The Kingdom of Saudi Arabia generates an enormous amount of date palm waste, causing severe environmental concerns. Green and strong concrete is increasingly demanded due to low carbon footprints and better performance. In this research work, biochar derived from locally available agriculture waste  was used as an additive to produce high-strength and durable concrete. Mechanical properties such as compressive and flexural strength were evaluated at 7, 14, and 28 days for control and all other mixes containing biochar. In addition, the durability properties of the concrete samples for the mixes were investigated by performing electric resistivity and ultra-sonic pulse velocity testing. Finally, a SWOT  analysis was carried out to make strategic decisions about biochar’s use in concrete. The results demonstrated that the compressive strength of concrete increased to 28–29% with the addition of 0.75–1.5 wt% of biochar. Biochar-concrete containing 0.75 wt% of biochar showed 16% higher flexural strength than the control specimen. The high ultrasonic pulse velocity (UPV) values (>7.79 km/s) and low electrical resistivity (<22.4 kΩ-cm) of biochar-based concrete confirm that the addition of biochar resulted in high-quality concrete free from internal flaws, cracks, and better structural integrity. SWOT analysis indicated that biochar-based concrete possessed improved performance than ordinary concrete, is suitable for extreme environments, and has opportunities for circular economy and applications in various construction designs. However, cost and technical shortcomings in biochar production and biochar-concrete mix design are still challenging. Moreover, activities related to the processing and transportation of cement are also responsible for greenhouse gas emissions, which are considered a severe environmental threat. Concrete contributes to approximately 8% of the entire world’s production of CO2 during the production, processing, and preparation phase [2 emissions and environmental issues, it has become necessary to implement sustainable CO2 reduction methods relevant to cement-based materials (CBM) in the environment [The construction industry is one of the most rapidly growing industries globally. The substantial growth of the construction industry led to the increasing demand for concrete. The population in urban regions is likely to grow from around 3.4 billion in 2009 to 6.5 billion by 2050. It is estimated that the yearly concrete production withstood at 10,000 Mt and is expected to rise twice in the next 40 years [ironment ,8,9. TheBiowaste, including municipal, industrial, agricultural, and other forms, is widely generated worldwide. This waste can be dumped into landfills and cause severe environmental consequences . TransfoThe primary aim of the presented research is to study the impact of biochar derived from date palm fronds as an additive in concrete to produce high-strength and durable concrete. The control and all other biochar-containing mixes assessed compressive and flexural strengths after 7, 14, and 28 days. Additionally, the durability properties of the concrete samples for the mixes were evaluated by measuring their electric resistivity and ultrasonic pulse velocity. Finally, the SWOT analysis and techno-economic assessment of the biochar-concrete system were performed to provide detailed insight into date palm derived biochar’s potential application on a commercial level.Type-1 Ordinary Portland cement (OPC) from a Saudi cement factory was used as a primary binder in all concrete specimens . The cemA total of 6 mix ratios were established to calculate the impact of biochar in standard strength concrete; 16 cylinders and two beams were cast for each mix. Five mixes contain various dosages of biochar, and one mix consists of ordinary Portland Cement (OPC) only. Biochar was added to the mix at a rate of 0.25%, 0.50%, 0.75%, 1.00%, and 1.50 wt% of the overall volume of concrete , represeAfter 7, 14, and 28 days of water curing, the compressive strength of cylindrical specimens was determined in three-time intervals. From each mix, three specimens were tested on a compression testing machine (CTM) with a loading rate of 2.4 kN/s, and the average value was noted according to  Figure .Beams with (100 mm × 100 mm × 500 mm) were utilised to calculate the flexural strength of concrete beams after 7, 14, and 28 days of water curing. Two beam specimens from every mix were assessed on a flexural testing machine with a consistent loading rate of 0.05 kN/s, and the mean value was noted according to (ASTM C 78)  Figure .The electrical resistivity of concrete was determined using nondestructive test equipment RESIPOD as per (ASTM C1876) standard  Figure . This teThe ultrasound pulse velocity of concrete specimens from each mix was determined using a portable ultrasonic nondestructive digital indicating tester (PUNDIT), according to (ASTM C597) standard , after 2The flexural strength biochar concrete specimens are displayed in In contrast to destructive testing, the research group focused on determining the mechanical properties of biochar-based concrete using non-destructive testing. ρ decreases gradually with increased biochar content. The value of electric resistivity was recorded as 26.1 kΩ-cm for the control mix. The values of electric resistivity noticed for 0.25%0.50%,0.75%,1.00%,1.50% biochar-concrete specimen was 22.4,22.2, 20.9,19.6,16.5kΩ-cm, respectively, which indicate linear decrease in the values of electric resistivity. Any material’s electrical resistivity (ρ) is described as its ability to resist the ions transfer exposed to an electrical field. It mainly relies on the microstructure elements associated with the shape of interconnection and pore size [The electrical resistivity of biochar-concrete and the control specimen is displayed in ore size . A finerore size ,41,42.Concrete is considered an essential building material globally and widely used for various construction applications. However, concrete manufacturing accounts for substantial greenhouse gas emissions associated with cement production. Therefore, innovative approaches toward green concrete building materials reduce environmental and climate impact and promote sustainable societal development. Recently, biochar-based concrete gained increasing attention due to its sustainability and improved mechanical and durable properties compared to ordinary Portland concrete ,43,44,45ρ), which is 47% lesser than the control concrete, suggesting that the biochar-concrete matrix consists of a heterogeneous structure with a strongly connected pore network.The major strengths of using date palm fronds derived biochar-based concrete as building materials are listed in −6 m2/s, respectively [2 emissions of the concrete industry. The study evaluated the impact of various governing factors, including raw materials, methods, synthesis, and transportation of biochar-concrete systems on the environment. It was estimated that approximately 0 to 20 wt% of biochar additions might expect to reduce 0.15–0.20 kg of cement in concrete. Therefore, using low cement amounts in biochar-concrete may be expected to reduce greenhouse gas emissions, ozone depletion, climate change, and hazardous biowaste management [Additionally, studies also confirmed that the high thermal stability of biochar-based cement composites is another essential factor demonstrating its applicability compared to ordinary concrete. It was reported that biochar-based mortar specimens consisting of different proportions  when subjected to different heating environments , showed minimal % loss in strength compared to ordinary mortar . The stuectively ,49. TherWhile other agricultural wastes such as rice husk ash, bagasse ash, palm oil fuel ash, etc., have been widely used to replace OPC in cementitious materials, there is little knowledge and availability on the properties of biochar made from date palm fronds and cementitious materials from it in most regions of the world. As a result of this lack of understanding, date palm fronds biochar application is very limited in the construction industry due to the lack of high confidence in the material. Furthermore, the effect of biochar on composite cement performance still requires various experiments to be completed to draw a more accurate conclusion. It is imperative to conduct an imminent study that examines the properties of date palm fronds biochar and its impact on cementitious materials’ long-term performance to promote the practical application of date palm fronds.The use of date palm fronds biochar in cementitious materials also has a weakness: its lower strength at high volume percentages. However, higher date palm fronds biochar content (1.5%) in the cementitious mix reduces its engineering performance, limiting its application as a binder component in cementitious materials ,52. ConvSimilarly, cementitious materials that contain date palm fronds biochar have a longer setting time, making them less suitable for applications requiring shorter setting times. A chemical additive such as an accelerator can be added to cementitious materials incorporating DPFA as a replacement for OPC to shorten their initial and final set times .Growing sustainability awareness in the construction industry has led to the search for sustainable materials that can replace OPC in cementitious materials. Due to its chemical properties and the fact that it is derived from agricultural waste, date palm frond biochar is an excellent alternative source of revenue for developing sustainable concrete. Saudi Arabia ranks among the world’s leading date-producing countries. As a result of the high production of dates, the date palm industry produces a tremendous amount of agricultural waste. If these wastes are improperly disposed of in the environment, they could pose a fire and safety risk. In addition, valuable land spaces could be depleted, and the aesthetics of the environment might be impacted. The processing of these date palm fronds into biochar would provide an opportunity to efficiently manage these waste materials and use them as a raw material for making green concrete. Further, converting these wastes into valuable resources would entail a monetary value for date palm factories, opening up another source of revenue.2 directly from the atmosphere in building materials and the addition of CO2-saturated biochar have a vital role. Even with a lack of studies on this topic, building materials that contain date palm fronds derived from biochar have superb possibilities for reducing carbon footprints and mitigating climate change [The application of date palm fronds biochar in building materials has been shown to improve the mechanical properties of building materials and enhance the durability of composites under extreme environmental conditions. The improved properties suggest that biochar-containing building materials perform equally or even better than those without ,59,60,61e change .Various potential barriers that may restrict date palm derived biochar-concrete commercialisation of biochar-concrete are listed in 2-eq./kg in the biochar life cycle using different biomass was estimated. In addition to reducing the net emission of GHG, the use of biochar in concrete has played a vital role in improving its chemical and mechanical properties. Most studies in the past showed significant improvements in concrete’s compressive and flexural strength [2-eq./kg as well as the generation of other value-added products such as syngas, bio-oil production in pyrolysis.The possible emissions of GHGs to the environment because of the decay/decomposition of the biomass are avoided by the process of valorising the biomass to produce biochar. Reducing GHG emissions of COstrength ,66,67,68strength ,64,69,70strength , compareApart from the environmental and other technical benefits of using biochar in concrete, its economic viability is crucial in deploying it in the construction industry. Kung et al.  reportedThe compressive strength of biochar-concrete increased with increasing biochar content and showed a maximum 28%, 26%and 29% improvement in power at 28-day age with the incorporation of 0.75%, 1.00%, and 1.50% of biochar. The biochar-concrete containing 0.75 wt% biochar loading indicated 16% higher flexural strength than the control mix. The increased surface area, small particle size, and water retention capability of porous biochar lead to a denser concrete matrix, formation of cement hydrates, and filler effect resulting in stronger concrete.Biochar-concrete showed high values (>7.79 km/s) of UPV demonstrating high-performance concrete. The electrical resistivity reduced linearly with the incorporation of biochar. This confirmed the formation homogeneous and denser biochar-concrete network resulting in lower permeable concrete.The SWOT and techno-economic assessment analysis further corroborates that the biochar-concrete system possessed the high potential to be commercially adopted as green and sustainable material despite the economic and engineering challenges.In general, it is suggested that biochar derived from Saudi agriculture waste can be used as a beneficial product for infrastructure designs requiring high-performance and durable building materials to attain technical and environmental benefits.In this research, biochar derived from date palm waste was used as an additive to concrete at the different mass compositions of 0.25 wt% to 1.5 wt%. The performance of biochar-concrete specimens derived from date palm waste was examined by the fresh concrete specimen’s representative mechanical and durability characteristics. The following conclusions can be drawn based on the outcomes:


The filter mud in molasses has a significant inhibitory effect on biological activity and cannot be utilised by organisms; therefore, before molasses are biotransformed, the filter mud will be separated and directly discarded in the environment. In this study, the filter mud was used as the retarder of cement concrete OPC 42.5 for the first time. It was found that when 0.2–0.8% filter mud was added to fresh cement concrete OPC PC 42.5, the hardening time of cement slurry was significantly prolonged due to the synergistic retarding effect of sugar, colloid and total cellulose in the filter mud. In addition, the compressive strength of cement concrete mixed with the filter mud in the early stage (<10 days), middle stage (10–100 days) and later stage (180 days) was significantly higher than that of cement concrete and cement concrete mixed with commercial asphalt lignosulfonate. These results showed that the filter mud in molasses could realise harmless and resource utilisation, which could promote the comprehensive utilisation of molasses. With the progress of hydration, the unstable clathrate will automatically decompose, and hydration will continue; (2) -OH, -NH2, -COOH groups are easily associated with water molecules through hydrogen bonding, coupled with the hydrogen bond association between water molecules, and a solvated water film will be formed on the surface of cement particles [2− on the surface of hydration products [The main reason for cement setting and hardening was that mineral compositions, dicalcium silicate 2CaO·SiOH), etc. . These hilicate CS and triH), etc. ,5,6. Ino C-A-S-H , therebyarticles ; thus, tproducts , coupledproducts ; thus, tproducts ,10,11.2 and C-S-H, thus hindering the setting and hardening of fresh concrete [Sugar and sugar derivatives contain active -OH groups and can be converted into saccharic acid containing -COOH in an alkaline solution . Sugar aconcrete ,14.Molasses is a cheap waste of biomass, and it is commonly used for the biosynthesis of many chemical products; however, molasses was often solid–liquid separated first, then the sugar solution with high sugar concentration was utilised , and theIn this paper, filter mud, a significant biological inhibitory component in molasses, was successfully utilised as a resource for the first time, that is, it was used as a cement retarder, which was conducive to increasing the types of molasses-based products and improving the comprehensive utilisation rate of molasses.Ordinary Portland cement OPC PC 42.5 was purchased from Shandong Linyi San’en Import and Export Co., Ltd. . The physical properties and chemical composition of the cement are given in Two kinds of crushed particles, mainly consisting of dolomite were used as aggregates: aggregate 1 (maximum particle size 16 mm) and aggregate 2 (maximum particle size 38 mm). Natural sand (maximum particle size 2 mm) and crushed stone sand (maximum particle size 4 mm) were used as fine aggregates. The volume proportions of various components in the aggregate mixture were 26% of aggregate 1, 28% of aggregate 2, 36% of natural sand, and 10% of crushed stone sand.w/w) solution. Formaldehyde was added to the mixture to prevent fermentation.Admixtures include Ca-lignosulfonate, molasses raw materials, filter mud from molasses, molasses colloid, sugar solution and water-soluble pigment. The molasses raw materials were purchased from Shandong Jinan Sugar Company ; the filter mud from molasses, colloid, sugar solution and water-soluble pigment was obtained by a gradual separation of the molasses raw materials (as shown in W/C (W/C: mass ratio of water to cement per cubic meter of concrete mixture) for all concrete mixtures were the same: 320 kg·m−3 and 0.65, respectively. The weight ratio of cement/aggregate 2/aggregate 1/crushed sand/natural sand was 1:1.7:1.6:0.6:2.0. The mixture was mixed in a 6 L mixer for the concrete test  for 3 min.The weight ratio of cement/sand/water in the mortar was 1:2.5:0.5. The cement content and The initial setting time and final setting time of cement concrete were measured by a Vicat apparatus  according to the method specified in GB/T1346-2011. The sample of cement concrete was cured in a moisture curing case , and the measurement was carried out 30 min after the sample was added to water. According to the methods specified in GB/T 50081-2002 and GB/T 50080-2002, the slump of the fresh concrete sample was measured by a concrete slump tester .WV is the total bled water/g, W is the water consumption of fresh concrete/g, G is the total fresh concrete/g, and GW is the mass of sample/g.The seepage characteristics of fresh concrete were analysed according to the bleeding rate. The test was carried out in the following method: A 5 L cylinder (with an inner diameter of 185 mm and a height of 200 mm) with a cover was dampened by a wet cloth; the fresh concrete was loaded into the cylinder one time and vibrated on a vibrating table; the surface of the cylinder was plastered with a trowel, and the cover was affixed to prevent evaporation. The surface of the sample was approximately 20 mm lower than the edge of the cylinder. From the time after the plastering operation, the bled water was sucked with a suction tube one time every 10 min in the first 60 min and one time every 20 min thereafter until three successive sucking operations had no water available. In the first 5 min before every sucking operation, one side of the bottom was underlaid up approximately 20 mm to make the cylinder tilt for water suction. After the sucking operations, the cylinder was gently placed flat and covered. The water obtained was fed into a graduated cylinder with a plug, the total bled water was measured, and the bleeding rate B was calculated according to Formula (1).3 at the ages of 1 d, 3 d, 7 d, 28 d, 90 d, 180 d and 200 d. The sample surface was compressed by a loading rate of 0.3–0.8 MPa·s−1 until the sample was near damage and started to deform, and the failure load was recorded.Compressive strength tests were carried out on cubic specimens of 15 × 15 × 15 cm3 cut from concrete specimens mentioned above on the 180th day to determine the freezing–thawing and wetting-drying properties. The sample was preserved for 28 days in saturated lime water and then in a laboratory with a relative humidity of 65 ± 5% and a temperature of 22 ± 2 °C. The sample was frozen in air, thawed and wetted in water. A wetting-drying cycle of 24 h was conducted. The sample was dried for 4 h in air at a temperature of 22 °C and then dried for 20 h in a baking oven at a temperature of 50 ± 2 °C. The degree of qualitative change in cement concrete should be determined by resonance frequency tests, ultrasonic velocity tests, and weight loss tests. A carbonation test was conducted for the fresh concrete sample after hardening for 28 days in the conditions with a relative humidity of 65 ± 5% and a temperature of 22 ± 2 °C, with phenolphthalein used as a lime indicator.Capillary tests were conducted for cubic specimens of 10 × 10 × 10 cmEach of the above tests was conducted three times in parallel, and related data were recorded as the average ± standard deviation.The setting time of fresh cement is given in 6-C3, sulfonic acid and other groups provide it with the structural characteristics of surfactant; thus, it shows certain surface physical and chemical properties, such as adsorption and dispersity, viscosity, rheological properties, and colloidal properties; however, it was a three-dimensional net structure of unsulfonated phenylpropane in the central site of the lignosulfonate molecule, and this kind of three-dimensional molecular structure hinders lignosulfonate from dispersing in cement; it was also detrimental to the adsorption of lignosulfonate on the surface of cement particles and failed to provide good adsorption retention and high surface activity, which caused poor compatibility between lignosulfonate and Portland cement [2 and on C-S-H gels, thereby preventing cement from setting [Lignosulfonate is a commonly used cement retarder and water reducer, and its phenolic hydroxyl group and hydroxyl group participate in multiple chemical reactions and physical interactions in cement retardation. Hydrophobic frame Cd cement . Sugar sd cement . These sd cement . When or setting ; howeverW/C value was, the more pores in cement were to be filled; that is, more demand for solid hydrates results in a longer time for cement setting and hardening. When the retarder was added, the setting time was further increased, and the greater the addition amount was, the longer the setting time was [2; thus, the hydration process was postponed. In fact, the retarding effect of celluloses could be attributed to the combined effect of the above two theories [W/C and the added dose of admixtures were the same, the initial setting time and final setting time of cement with added filter mud from molasses advanced; however, increasing the amount of filter mud from molasses could reach the same effect as adding molasses raw materials, and in practical applications, the amount of filter mud from molasses could be selected as required; therefore, filter mud from molasses could still be considered an effective, promising cement retarder and a water reducer that could replace lignosulfonate, and it could achieve the purpose of making full use of molasses raw materials.When the addition amount exceeds the critical dose, the setting time could rapidly fall from the maximum until the coagulating effect occurs  because time was . When cotime was . The moltime was . Filter theories . In thisThe slump of the fresh concrete sample is given in W/C value and additional amount of retarder, the concrete with added lignosulfonate had the largest slump, that with added filter mud from molasses had the second largest slump, and that with added molasses raw materials had the smallest slump, which was probably attributed to the higher air content in fresh concrete with added lignosulfonate and that with added filter mud from molasses. Jumadurdiyev et al. [As shown in v et al.  comparedv et al. . The sliv et al. ,22.The bleeding rates of fresh concrete with added molasses raw materials, filter mud from molasses, and lignosulfonate are given in The influences of molasses raw materials, filter mud from molasses and lignin sulfonate on the compressive strength of hardened concrete are given in Q is the cumulative water absorption per unit area, mm; a is a constant, min; and K is the absorbance.Water absorption of concrete with 0.4% molasses, 0.4% molasses and lignosulfonate was observed (as shown in W/C value was the same, the concrete with added lignosulfonate and that with added filter mud from molasses had poorer water absorption capacity than that with added molasses, which was attributed to the higher air content in the concrete with added lignosulfonate and that with added filter mud from molasses. In most cases, bubbles in the concrete reduced the number of pores and disturbed the continuity of capillary pores in the cement. In addition, the cement concrete supplemented with lignosulfonate and that supplemented with molasses showed a lower bleeding rate than that supplemented with molasses; this was because the concrete with added lignosulfonate and that with added molasses had poorer water absorption capacity than that with added molasses.As shown in The results of the drying shrinkage test of concrete at the age of 0 to 180 days are given in W/C values and found that the concrete with W/C equal to or higher than 0.60 had good freezing–thawing resistance, and the concrete with added sugar had better freezing–thawing resistance than that without sugar; however, after 100 cycles, the measured longitudinal ultrasonic velocity showed that the cement concrete sample had suffered slight deterioration (The variation in resonant frequency in the freezing–thawing test was 100 cycles. As shown in ioration . The velAfter 60 cycles of the wetting and drying test, the concrete sample obtained resonant frequency and ultrasonic velocity, as shown in The carbonation depth of the concrete with added molasses raw materials, that with added filter mud from molasses and that with added lignin sulfonate is shown in The main conclusions can be summarised from this work as follows. Molasses filter mud could be used as a good substitute for lignosulfonate, and could also partially replace molasses as retarder and water reducer of OPC. The molasses filter mud-added cement pastes showed expanded setting times even in 0.2% dosage; however, the water-soluble pigment in molasses had no significant effect on the setting time of cement paste, and colloids significantly promoted the setting of cement paste. In addition, the filter mud significantly reduced the slump of cement concrete and increased the compressive strength and water absorption capacity of OPC. After 100 freeze–thaw cycles, the concrete with molasses filter mud had better freeze-thaw resistance than the concrete with lignosulfonate; therefore, the performance of filter mud in concrete workability was similar to that of lignosulfonate-based admixture, which provided a potential way for the comprehensive utilisation of molasses.


Background: The possible relation between antibiotic exposure and the alteration of gut microbiota, which may affect dementia risk, has been revealed. However, the association between antibiotics and dementia incidence has rarely been studied. We aimed to determine the association between antibiotic exposure and the risk of dementia.Methods: This population-based retrospective cohort study used data from the National Health Insurance Service-Health Screening Cohort (NHIS-HEALS) in South Korea. Exposure was the cumulative days of antibiotic prescription from 2002 to 2005. Newly diagnosed overall dementia, Alzheimer’s disease (AD), and vascular dementia (VD) were identified based on diagnostic codes and prescriptions for dementia-related drugs. The follow-up investigation was carried out from 1 January 2006 to 31 December 2013. The Cox proportional hazards regression was used to assess the association between cumulative antibiotic prescription days and dementia incidence.Results: A total of 313,161 participants were analyzed in this study. Compared to antibiotic non-users, the participants who used antibiotics for 91 or more days had an increased risk of overall dementia , AD , and VD . Those who used five or more antibiotic classes had higher risks of overall dementia  and AD  than antibiotic non-users.Conclusion: Antibiotic exposure may increase the risk of dementia in a cumulative duration-dependent manner among adult participants. Future studies are needed to assess the causality between the long-term prescription of antibiotics and dementia risk. Dementia is one of the most prevalent neurodegenerative diseases globally . In 2020Gut microbiota consists of the aggregate of all microorganisms that affect essential human functions . SpecifiPrevious studies about the relationship between antibiotics and dementia are insufficient. The gut microbiome is highly affected by an unhealthy lifestyle  and its significant changes have been reported in patients with AD . In partThis population-based retrospective cohort study was conducted to assess the association between antibiotic exposure and the risk of dementia incidence using the Korean National Health Insurance Service-Health Screening Cohort (NHIS-HEALS) database.The NHIS provides compulsory health insurance for all South Korean, and it covers almost all types of medical services, including health screening for all dependents aged 40 and over biennially. The NHIS-HEALS database is constructed by a 10% random sampling of all individuals aged between 40–79 years who participated in the 2002 and 2003 health examinations, and these participants were followed up until 2013 . The datAmong the 334,265 participants who completed health screening between 2004 and 2005, 858 individuals who died before the index date of 1 January 2006, were excluded. In addition, we excluded 1,290 individuals who had a dementia diagnosis or a prescription of dementia-related drugs before the index date. The 18,956 individuals who had missing covariate values were also excluded . AntibioThe primary outcome was the development of dementia during the observation period, defined by the 10th Revision of the International Classification of Diseases (ICD-10) codes of F00–F03 and G30 with prescription records of anticholinesterase drugs .The diagnosis codes for Alzheimer’s disease  and vascular dementia  were adopted from a previous study .The exposure variables were the cumulative days of antibiotic prescription and the number of antibiotic classes between 2002 and 2005, the first 4 years in the database. The cumulative days of antibiotic use were calculated as the sum of all days of antibiotic subscription from 2002 to 2005. Penicillin, cephalosporin, macrolides, fluoroquinolones, sulfonamides, lincosamides, tetracyclines, and vancomycin were defined by the World Health Organization Anatomical Therapeutic Chemical (ATC) classification of drugs. Antibiotic exposure groups were classified as 0, 1–30, 31–90, and 91 or more days of prescriptions. The number of antibiotic classes was classified as 0, 1, 2, 3, 4, and 5 or more.Covariates were extracted between 2002 and 2005, including age, sex, body mass index (BMI), smoking status, alcohol consumption, physical activity, household income, Charlson comorbidity index (CCI), fasting blood sugar, systolic blood pressure, total cholesterol, infectious diseases  , and antMultivariable Cox proportional hazard regression was performed to estimate the adjusted hazard ratios (aHRs) and 95% confidence intervals (CI) for overall dementia, AD, and VD by antibiotic use. Age, sex, BMI, smoking status, alcohol consumption, physical activity, household income, CCI, fasting blood sugar, systolic blood pressure, total cholesterol, and antidepressant use were considered as potential confounders in the first model. The antibiotic non-user group was considered the reference group in the first model. The second model additionally adjusted for infectious diseases, which are the primary cause of antibiotic usage. In addition, 1–30 days of the exposure group was used as the reference group when evaluating the risk of dementia among antibiotic users only. Stratified analyses of subgroups that were stratified by age, sex, BMI, physical activity, household income, CCI, and antidepressant use were performed. Furthermore, we carried out sensitivity analyses that determined the association of antibiotic use with dementia risk after excluding patients diagnosed with dementia within the first 1–3 years of follow-up. In other words, we did not consider new cases of dementia for those diagnosed within the first 3 years of follow-up to address protopathic bias . We alsop-value <0.05 in a two-sided manner. P for trend was calculated by the cumulative days of antibiotic exposure and the number of antibiotic classes, as a continuous variable, separately. P for interaction in Cox proportional hazards models was calculated by including interaction terms between antibiotic exposure and covariates.Data collection, data mining, and statistical analyses were conducted using SAS 9.4 , STATA ver. 13.0 , and R version 4.2.0 . Statistical significance was defined as a The baseline characteristics of participants are shown in The association between cumulative antibiotic prescription days and the risk of developing overall dementia, AD, and VD is depicted in In The effect of the number of antibiotic classes prescribed on dementia incidence is demonstrated in In this nationally representative cohort study, antibiotic use was associated with the risk of dementia incidence. There was a cumulative duration-dependent relationship between dementia incidences with cumulative days of antibiotic exposure. The results were significant after adjustments for all covariates. Furthermore, the results of the propensity score matching analysis verified our primary findings. To our knowledge, this is the first study to indicate that longer cumulative days of antibiotic exposure could lead to significantly higher dementia risk.Previous studies showed that antibiotics can induce alterations in the gut microbiota, leading to AD . In a ChDementia is a chronic neurodegenerative disease with clinical characteristics such as slow onset and gradual progression . In partAR), causing central excitotoxicity (Multiple mechanisms of the association between antibiotics and dementia risk have been postulated. First, the gut microbiota–brain axis is considered a crucial way that mediates the relationship between gut dysbiosis by antibiotics and dementia. A recent study has suggested that antibiotics can cause neuroinflammation and neurodegeneration through the gut microbiota–brain axis, favoring AD or worsening its course . Anothertoxicity . In our toxicity . It is btoxicity . Furthertoxicity . Thus, gOur study had several limitations. First, the study population was confined to participants who underwent a national health examination. There is a possibility of selection bias that participants in a bad health have been excluded. Second, the operational definition of dementia was based on the claim data. Therefore, it has a validation problem about the real diagnoses although we defined the outcome by diagnosis and prescription concurrently. Thus, future studies with medical chart reviews are required. Another limitation concerns the follow-up duration of 8 years, which may not be long enough to fully disclose the effects of antibiotics on the risk of dementia. As dementia usually develops slowly with gradual cognitive decline , future Despite these limitations, our study’s strengths include the ability to assess the association of antibiotic exposure with dementia risk in older adults. First, the NHIS-HEALS database was based on the medical claim records collected at the nationally representative level. It contains individual-level information about demographic data, health examinations, diagnoses, and prescriptions. Furthermore, a large study population of 313,161 may strengthen the generalization of our results. Second, we conducted the association between antibiotic exposure and dementia incidence by regarding various risk factors such as age, household income, CCI, and infectious diseases. Third, we conducted sensitivity analysis and propensity score matching analysis to determine the robustness of our findings. Analyses that considered latent periods to address reverse causality also showed consistent results.Antibiotic exposure appears to be associated with greater dementia incidence in our cohort study. There was a cumulative duration-dependent association between antibiotic exposure and dementia risk. However, it is possible that individuals who were long-term prescribed antibiotics had a poor health status, which increases their vulnerability to dementia. In other words, antibiotic exposure might be a proxy for dementia risk factors that we could not consider in this study, not a direct cause of dementia. This study suggests that antibiotic exposure could be one of several risk factors for dementia. Ultimately, future prospective and epidemiological studies with a broader study population that investigate the association between changes in gut microbiota by antibiotic exposure and dementia incidence are needed to validate the interpretation of our findings.


We believe that major steps can be taken towards Acquired Immunodeficiency Syndrome (AIDS) prevention through identifying the relevant factors that are apt to predict risky behavior. The main purpose of the present study was to analyze and evaluate the relationship of social support, family functioning, self-efficacy and AIDS risk perception to controlling risky behavior associated with AIDS.To conduct this cross-sectional study, 765 subjects  were selected from the youth inhabiting the western provinces of Iran through cluster sampling. Five questionnaires were used: AIDS risk perception, self-efficacy in controlling risky behavior associated with AIDS, controlling risky behavior associated with AIDS, the multidimensional scale of perceived social support, and the family assessment device.p < 0.01).The results demonstrated that all two models enjoyed acceptable fitness, and the mediating roles of self-efficacy and AIDS risk perception were confirmed. Moreover, family functioning and perceived social support together could predict 20% of the variance of controlling risky behavior associated with AIDS. The results also indicated that family functioning with a standardized coefficient of − 0.24 and self-efficacy in controlling risky behavior associated with AIDS with a standardized coefficient of 0.58 could predict the controlling risky behavior associated with AIDS (Our findings suggest that self-efficacy and AIDS risk perception play major roles in controlling risky behavior associated with AIDS. Therefore, it is recommended that families and psychologists promote self-efficacy in order to prevent the occurrence of high-risk behaviors. AIDS is still considered as one of the most prominent health problems worldwide in a way that the statistics announced in 2007 indicated that the various rates of AIDS infection were as follows: 3–7.2 million annually , given tOne can change one’s behavior. In this regard, Albert Bandura (1986) considers self-efficacy as a major factor in changing behavior , and accSeveral theories about health-promoting behaviors have already been proposed by researchers, which were also considered in this study, including social-cognitive theory , protect). The inclusion criteria were to be in the 18–35 age range, a five-year residency in these provinces at the least, and studying for at least six whole academic years and people who did not want to collaborate in the study were excluded. Samples were selected using cluster sampling. To do so, three provinces  were randomly chosen from five western provinces in Iran. Then, 250 subjects were selected from each of three provincial capitals according to the municipal zoning using Cochran's sample size formula. Additionally, given the likelihood of anticipated sample dropouts and lack of cooperation, 800 questionnaires were distributed among the target subjects residing in the three provincial capitals. To this end, the researchers walked around the cities, detailed explanations of how to complete the questionnaires were supplied, and the participants were requested to ask for more clarification in case of encountering problems filling out the questionnaires. Additionally, they were assured that their information would remain confidential, and their informed consent was taken. Further, the questionnaires were completed individually and collectively in the presence of the researchers. Finally, 765 questionnaires were collected and then analyzed.The statistical population of the present cross-sectional study consisted of the whole youth inhabiting the western provinces of Iran in 2016 .This 13-item tool was scored on a five-point Likert scale . The results of analyzing the reliability of the questionnaire demonstrated that there was a 0.70 correlation coefficient between the first and second stages of test–retest reliability, and Cronbach's alpha was 0.70. Moreover, the validity and reliability of this tool have been confirmed . The resThis 13-item tool was scored on a six-point Likert scale . The results of analyzing the reliability of the questionnaire indicated that there was a 0.91 correlation coefficient between the first and second stages of test–retest reliability, and Cronbach's alpha was 0.85. Moreover, the validity and reliability of this tool have been confirmed , and itsThis 14-item tool was scored on a six-point Likert scale . The results of analyzing the reliability of the questionnaire showed that there was a 0.77 correlation coefficient between the first and second stages of test–retest reliability, and Cronbach's alpha was 0.77. Further, the validity and reliability of this tool have been confirmed , and itsIn this 12-item scale, developed by Zimet et al. (1990), three dimensions of family support, support from friends, and support from others are measured . FurtherThis 60-item tool, formulated based on McMaster’s pattern, determines the structural, occupational, and interactive characteristics of families based on McMaster’s pattern. Further, six dimensions of family functioning are specified in this tool: problem solving, communication, roles, affective responses, affective involvement, and behavior control. It also includes a subscale pertaining to the general functioning of families. Scoring is based on Likert scale, and lower scores are indicative of healthier functioning. Further, a reliability of 0.72 to 0.92 has been reported in this questionnaire , and itsThe obtained data are analyzed using SPSS-18, SPSS Inc, Chicago, Illinois, USA and AMOS-18 software application and structural equation modelling method (SEM) based on the variables of the model. Based on the obtained model, we can foresee which variables have the capability of predicting the risky behaviors. Two models were assessed in the present study: AIDS risk perception and self-efficacy in controlling risky behavior associated with AIDS were examined in the first and second model.The current correlational study was a structural equation modeling. Descriptive and inferential statistical methods were used in the present study. These methods include mean and standard deviation method to describe the status of variables, simple correlation coefficient (Pearson) between variables to examine the relationship between variables and related test hypotheses (such as the linearity of the relationship between independent and dependent variables), other methods include statistical analysis to study structural equation modeling assumptions, structural equation modeling method to test the models in the present research, Macro bootstraping analysis of indirect relationships to investigate indirect effect of family functioning and social support in controlling risky behavior associated with AIDS. Data analysis was performed using SPSS-19 and amos-19 statistical software.Of the 765 participants who entered the study, 59% (448 subjects) were female, and the average age of the entire sample was 27.5 ± 92.33. Socio- demographic information of participants are shown in Table The mean and standard deviation of the main variables of the study as well as ages in each gender are shown in Table As shown in Table The matrix of correlation coefficients between the variables under study is shown in Table p < 0.001) and perceived social support with a standardized coefficient of 0.17 (p < 0.02) could predict the variable of controlling risky behavior associated with AIDS. To investigate the mediating role of AIDS risk perception in the relationship of family functioning and perceived social support to controlling risky behavior associated with AIDS, a model was formulated.First, a model was designed in the absence of the mediating variables. The results demonstrated that family functioning and perceived social support together could predict 20% of the variance of controlling risky behavior associated with AIDS. In addition, the results showed that family functioning with a standardized coefficient of − 0.30 (p < 0.009), perceived social support with a standardized coefficient of 0.13 (p < 0.04) and AIDS risk perception with a standardized coefficient of 0.47 (p < 0.01) could predict the variable of controlling risky behavior associated with AIDS.The results of analyzing the model displayed in Fig. The results demonstrated that the variables of family functioning, perceived social support and AIDS risk perception had direct effects on controlling risky behavior associated with AIDS, whereas only family functioning (β = − 0.13) had indirect effects on controlling risky behavior associated with AIDS  and self-efficacy in controlling risky behavior associated with AIDS with a standardized coefficient of 0.58 (p < 0.01) could predict the variable of controlling risky behavior associated with AIDS.The results of analyzing the model displayed in Fig. The results indicated that the variables of family functioning and self-efficacy in controlling risky behavior associated with AIDS had direct effects on controlling risky behavior associated with AIDS, whereas perceived social support did not have direct effects on controlling risky behavior associated with AIDS. Moreover, the results showed that family functioning did not have indirect effects on controlling risky behavior associated with AIDS  denote unhealthy functioning, i.e., the healthier the family functioning, the higher the perception of the members of the family of AIDS, they will have higher self-efficacy in controlling risky behavior and controlling risky behavior associated with AIDS are better conducted. Furthermore, the results of the present study showed that family functioning could predict the risky behavior associated with AIDS and AIDS risk perception, whereas it could not predict self-efficacy in controlling risky behavior associated with AIDS. Previous studies conducted in this field have been indicative of the role of family in displaying risky behavior associated with AIDS, including drug use and risky sexual behavior –21. So, The results of a systematic study demonstrated that the role of family structure has been confirmed in risky behavior in studies conducted in African countries over 2000–2014 .However, some comparative studies showed that there were differences between the criminals’ families and peer groups , 47. SomIn another section, the results of the present study demonstrated that perceived social support, AIDS risk perception, self-efficacy in controlling risky behavior associated with AIDS and controlling risky behavior associated with AIDS were positively related. In other words, the higher one’s perceived social support, the extent of each of the other three factors will be greater.As a further matter, the results indicated that perceived social support and family functioning together could predict controlling risky behavior associated with AIDS, but its impact factor was less than that of family functioning. The results of this section of the study were consistent with the preceding studies , 8, 9. AMoreover, the results of the present study showed that AIDS risk perception and controlling risky behavior associated with AIDS were positively related, i.e., higher risk perception will result in higher control over risky behavior. In the proposed model for predicting risky behavior based on family functioning and perceived social support, AIDS risk perception played a mediating role. In other words, it can be said that proper family functioning and high social support will lead to controlling risky behavior when one’s AIDS risk perception is high. The results of various studies have been indicative of the fact that low AIDS risk perception will give rise to risky sexual behavior , 23, 54,As a further matter, in the proposed model for predicting risky behavior based on family functioning and perceived social support, it was concluded that self-efficacy played a mediating role. The relationship between self-efficacy in controlling risky behavior associated with AIDS and AIDS preventive behavior has been confirmed in preceding studies, and this construct can play a major role in decreasing risky behavior leading to AIDS –31. The Furthermore, gender is one of the variables that plays a key role in the risk of AIDS. Gender roles in any society may organize relationships that play a key role in AIDS . In the Iranian and Islamic cultures through religious beliefs, and teachings may influence the pattern of social relations especially in the AIDS/HIV disease. In Iran and other Islamic countries, there may be a belief that HIV is caused by extramarital relationships. Therefore, self-restraint from extramarital and amoral sexual behaviors is the important way to decrease the spread of AIDS/HIV. Moreover, Dadipoor (2020) claimed that AIDS is condemned among Iranian people . ADIS isHowever, in Iran, the role of cultural factors in prevention of AIDS are still in an early stage. It follows that sociocultural factors are also a new issue and needs more investigation. Given the lack of reliable information on the status of people with ADIS, there is a great need for research and most importantly for interventions to prevention and to cope with knowledge related to this disease. Therefore, those interventions have to be culturally sensitive.According to the results, it can be said that self-efficacy is a mediating variable in the relationship of family functioning and perceived social support to controlling risky behavior. Therefore, it is recommended that families and psychologists promote self-efficacy in order to prevent the occurrence of high-risk behaviors.Given that the present study was conducted in Iran, extreme caution must be exercised when generalizing the results and these models must be investigated in other societies, too. Hence, it is better to take this point into consideration when generalizing the results. Since our study was conducted on Iranian young’s, future research should be conducted on other samples. This study was unique and the results was interesting, so we can use the results across prevention of AIDS.


Saline soil in Western China contains high concentrations of chloride ions, sulfate ions, and other corrosive ions, and the performance of concrete will substantially deteriorate from exposure to this environment. Therefore, it is of great significance to study and predict the concrete compressive strength in saline soil environments. In this paper, the effects of corrosion on concrete were analyzed from the aspects of surface damage, damage depth, and X-ray diffraction (XRD) of the corrosion products. The effects of corrosion were quantified by damage depth and corrosion depth. Then, considering the corrosion effects combined with Fick’s diffusion law, a time-dependent model of concrete compressive strength and a time-dependent model of damage depth were established. The results show that the deterioration of concrete gradually developed from the surface to the interior, and that the interface of the concrete specimen was equivalent to three parts: a failure zone, a filling zone, and an undisturbed zone. The results also showed that the time-varying model of concrete compressive strength proposed by the author was fully applicable, with an error of less than five percent. The service life of concrete predicted by the damage depth was found to be about 253 months (21.1 years), and the service life predicted by the time-varying compressive strength model was about 187 months (15.6 years). Both prediction results were far less than the normal concrete service life of 50 years. In addition, the long-term compressive strength of the corroded concrete was about 90% of that of the noncorroded concrete, which did not deteriorate with the corrosion time. There3A)  in concr2·10H2O) ,10, whic2·10H2O) . The phyO4·10H2O ,13. The aterials . The gyp[4·2H2O)  and ettr4·31H2O)  are two 4·31H2O) . However4·31H2O) ,19, causIn view of the mechanism of concrete degradation in the abovementioned chloride and sulfate environments, many scholars have studied the compressive strength of concrete which has been exposed to corrosive environments. Some scholars have found that the compressive strength tends to gradually decrease after exposure to corrosive environments for different durations ,21, and −, SO42−, CO32−, Na+, K+, and Mg2+, are widely distributed in Northwest China [Saline soils, which contain high concentrations of Clst China ,30; thesst China ,32,33. Mst China . In this3. The size of the crushed stone was 5~25 mm, and its apparent density was 2660 kg/m3. The mix of concrete is shown in The test piece was a cubic specimen with side lengths of 100 mm. The total number of test pieces was 36. The chemical composition and the working performance index of the cement P. O. 42.5 R, according to Chinese Portland cement Standard , similarThe manufacturing process of the test piece is as follows: (1). Weigh the cement, stone, sand, and water used in the test according to mass mix of concrete in There were 6 dry and wet cycle inspection periods in total: 0 months , 5 months, 8 months, 10 months, 15 months, and 20 months. Six cubic specimens were soaked in each inspection period, with a total number of 36 specimens. The test solution was composed of the same composition and mixture ratio as saline soil in Western China [After the dry and wet cyclic corrosion test of each group of concrete specimens was completed, compressive tests were carried out on three of the concrete specimens. Subsequent analyses of the free chloride content and the free sulfate content and the XRD analysis of the corrosion products were carried out on the other three specimens.When testing the distribution of ions in concrete, the corresponding test specimens were ground and sampled. The sampling depth was within 20 mm from the concrete surface, and samples were taken every 2 mm. After exceeding 20 mm, samples were taken every 5 mm—that is, the sampling depths were 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 22.5, 27.5, and 32.5 mm, respectively. After the sample was taken, it was then passed through a 0.63 mm sieve to remove large particles. According to the relevant Chinese standard , the fre2CrO4 indicator with a concentration of about 5%, phenolphthalein solution with a concentration of about 0.5%, dilute sulfuric acid, and standard NaCl solution with a concentration of 0.02 mol/L. Prepare a V2 mL standard silver nitrate solution with a concentration of about 0.02 mol/L with a V1 mL standard NaCl solution. The calculation formula of its concentration CAgNO3 is shown in (1),Prepare K3) of distilled water, tighten the cork, and shake it violently for 1–2 min. Soak it for 24 h. Shake it again after soaking for 12 h to ensure that the chloride ions in the sample are fully dissolved in the distilled water.Place the concrete sample in the drying box oven for 2 h with a drying temperature of 105 ± 1 ℃ to ensure that the water in the sample is fully volatilized. Accurately weigh 2 g (recorded as G) of the dried sample and pour it into a triangular beaker. Add 200 mL (V4) portions of filtrate, respectively, and put them into two triangular flasks; add two drops of phenolphthalein to the two triangular flasks, respectively, and then neutralize them with dilute sulfuric acid until they are colorless to ensure that the filtrate is neutral. Add 10 drops of potassium chromate indicator into the triangular flasks, and then titrate them with silver nitrate solution immediately until brick red sediment appears. Record the volume of silver nitrate solution (V5) consumed (the triangular flask needs to be shaken violently during titration). The calculation formula of free chloride ions is:fc is the content of free chloride ions in the sample ; CAgNO3 is the concentration of silver nitrate solution (mol/L); G is sample mass (g); V3 is the volume of distilled water added to the sample (mL); V4 is the volume of filtrate used for each titration (mL); and V5 is the volume of silver nitrate solution consumed after titration (mL). The final measured value of fc is the average value of the two measured results.Use neutral medium-speed filter paper to filter out the sediment in the soaking solution; measure two 20 mL  mixture was prepared in advance, and then 2.0 g of concrete powder sample was weighed, soaked in 50 mL distilled water, shaken for 2 h, and then soaked for more than 24 h. The sample was filtered with slow-speed filter paper. An amount of 25 mL filtrate was taken into a 50 mL volumetric flask. First, 2.5 mL hydrochloric acid was added to the volumetric flask, followed by 10 mL of uniformly mixed BaCl2-PVA mixture. The volume of distilled water was fixed to 50 mL, and the turbidity of the solution was shaken 2–3 times by hand. After the turbidity of the solution was obviously stable, the solution was left to stand for 5 min and then tested by ultraviolet spectrophotometer. The relationship between the SO42- content measured in the test and the absorbance value is as follows:fsc is the mass of sulfate ions in the 50 mL solution (mg) and abs is the absorbance value displayed on the spectrophotometer display.Ultraviolet spectrophotometer method was used to measure the content of free sulfate ions, as shown in According to the absorbance value abs measured by the spectrophotometer, the free sulfate ion concentration can be calculated.3 coloration method [Assuming that all ions had the same damage depth in concrete during the same dry and wet cyclic corrosion duration, the damage depth was expressed by the chloride diffusion depth in this paper, and the chloride diffusion depth was determined by the AgNOn method .2O precipitation. An obvious color boundary, namely a color trace, formed at the junction of the two different color regions. Finally, vernier calipers were used to measure the depth of the color-developing area of the specimen section. Usually, the average width from the penetration surface to the color-changing boundary was measured as the average erosion depth of chloride ions.First, the corresponding specimen was cut from the middle part, and two planed sections were selected as the test surface. Then, 0.1 mol/L silver nitrate solution was sprayed evenly on the concrete section. In the area where the chloride ion concentration was greater than a certain critical value, the chloride ions reacted with the silver ions to form a white AgCl precipitation. When the chloride ion concentration was lower than a certain value, the hydroxyl ions reacted with the silver ions to form AgOH, and then oxidized to a brown AgAfter the dry–wet cycle erosion test was completed and the corresponding specimens were taken for microscopic observation and corrosion product analysis. First, the specimens were cut and sampled at the depth of 10~15 mm from the concrete surface. After the test sampling was completed, the coarse aggregate in the samples was removed with a 200 mesh sieve, and then the carbon spraying treatment was carried out. Then, the SEM-EDS test was carried out on a SIGMA 500 field emission scanning electron microscope manufactured by Zeiss AG of Jena, Germany. The magnification of the concrete SEM image was 2000 times. Finally, the change of the internal microstructure of the concrete specimen was analyzed through the test results. EDS analysis was carried out by X pert3 powder X-ray diffractometer to detect the types of elements contained in the sample and to study the corrosion products generated in the concrete after erosion. XRD test conditions: Target was Cu Target, acceleration voltage was 40 kV, current was 40 mA, and scanning angle was 5–80°.The electro-hydraulic servo universal testing machine, with a maximum range of 1000 kN, was used as the basic mechanical property test instrument of concrete. According to the relevant requirements in the national standards of China , the cubThe failure patterns and variations in the compressive strength after different dry and wet cycle inspection periods is shown in cuf is the compression strength (MPa), 0.95 is the size conversion factor, F is the failure load (N), and A is the area of the cross section (mm2).According to the relevant Chinese code , the comAs can be seen from The surface changes in concrete after different dry and wet cycle inspection periods are shown in 2SO4, and CaCO3. The test solution contained NaCl and Na2SO4, but did not contain Ca2+, indicating that the Ca2+ came from the concrete. The test solution reacted with Ca(OH)2 and C-S-H in the concrete to form soluble calcium-containing substances, which precipitated from concrete in the form of CaCO3. The contents of Ca(OH)2 and C-S-H gel in the concrete decreased due to the precipitation of calcium ions, resulting in decreased concrete strength.The XRD analysis was carried out on samples of the white crystalline substances from the “salting out” phenomenon on the concrete surface, and the results are shown in 3A only fixes chloride ions after an initial period, which is longer than the one corresponding to the absorption process. In contrast, the effect of the C4AF fixing chlorides by forming chloroaluminates is significant [The damage depths of the specimens after different dry and wet cycle inspection periods are shown in nificant . This finificant  reached 2O3·3CaSO4·32H2O), gypsum crystal (CaSO4·2H2O), Friedel’s salt (3CaO·AI2O3·CaCl2·10H2O), CaCO3 crystal, and other substances. The frontal angles of ettringite crystal are mainly 27°, 50°, and 60°. The frontal angles of gypsum crystal are mainly 32° and 36°. The frontal angles of Friedel’s salt are mainly 18°, 21°, 26°, 34°, and 39°. The frontal angles of CaCO3 crystal are mainly 29° and 47°. Both gypsum [- and the cement hydration product (C–S–H gel), has no cementitious properties; therefore, the formation of Friedel’s salt results in a decrease in the cementing ability of the concrete. The above analysis shows that the corrosion effect is the main reason that the compressive strength of concrete first increases and then decreases.The corrosion products of the concrete specimens after corrosion for 10 months were analyzed by XRD at the distance of 10~15 mm from the surface of the concrete, as shown in h gypsum  have typh gypsum . On the dx(t) and corrosion depth cx(t), as shown in As mentioned above, whether the corrosion effects fill or expand the internal pores of concrete depends mainly on the relationship between the content of the corrosion products and the internal porosity of the concrete . The disA and compressive strength of the concrete can be expressed as:It is assumed that the concrete is a homogeneous material. When the concrete is compressed, the concrete can be divided into N vertical rods. It was also assumed that the properties of each rod were the same, that the cross-sectional area was S, and that the failure stress was dA of the failure zone can be expressed as:In the failure zone, there were M bars failure. The stress that each rod can bear is cA of filling zone can be expressed as:In the filling zone, there were O bars strengthened. The stress that each rod can bear is The parallel bar model of compressive strength of corroded concrete considering corrosion effect can be expressed as:cuf(t) is the compressive strength of concrete after corrosion for t month; cuf(0) is the compressive strength of noncorroded concrete; dA(t) and cA(t) are the cross-sectional areas of the failure zone and filling zone, respectively; A is the total cross-sectional area of the cross section; and a is the length of cross section.In conjunction with the above formulas, Formula (10) can be obtained:dγ in this paper was 0.9.Various concrete specimens buried at Dun-huang Station in the saline soil area of Western China in 1959 were excavated and measured in 1995 after being buried in the soil for 36 years. After the corrosion of saline soil, the compressive strength of the ordinary Portland cement concrete tested in 1995 was approximately 10% lower than the original strength . Kwon 44 placed cf(t) is the compressive strength when the porosity is p(t), ω is the cement content of the concrete, and mr is the average pore size distribution. The average mr of the concrete specimens after different dry and wet cyclic corrosion durations measured in this paper was 20.4 nm.Kumar and Bhattacharjee  establisp(t) of the concrete specimens and the dry and wet cyclic corrosion duration t is as follows:The porosities of the concrete specimens taken 7–12 mm away from the surface of the concrete were measured, and the variation in the porosities after different dry and wet cycle inspection periods is shown in cγ can be expressed by Formula (10):The corrosion enhancement coefficient x(t) can be obtained as follows:x(t) is the diffusion depth when the dry and wet cycle inspection period is t months (mm), fc (x(t),t) is the free ion content at the depth of x(t) when the dry and wet cycle inspection period is t months (%), sc is the surface ion content (%), c0 is the initial ion content (%), D is the ion diffusion coefficient (mm2/s), and erf−1(u) is the Gaussian error inverse function.The diffusion of ions in concrete conforms to Fick’s diffusion law, which is essentially the diffusion of ions under the action of concentration difference . Therefoc0, i.e., t, the free ion content on the concrete surface is sc, i.e., t, at an infinite depth, the content of free ions in concrete is c0, i.e., c0 = 0 in this study. Hence, Equation (14) can be changed to:In Fick’s diffusion law , it is usc and the diffusion coefficient D of chloride were obtained, as shown in The distribution of free chloride and free sulfate content versus the diffusion depth is shown in 3 color coloration method. The free chloride content here was defined as the damage depth. It was found that the free chloride content here accounts for 0.15~0.28% of the concrete quality. Therefore, dx(t).Some scholars ,49 studicx(t).Carlos  investigsc and diffusion coefficient D of chloride in dx(t) and corrosion depth cx(t) of concrete specimens at different dry and wet cyclic corrosion times could be obtained.When dx(t), the corrosion depth cx(t), the corrosion damage coefficient dγ, and the corrosion enhancement coefficient cγ into Formula (10), and a comparison between the calculated and test values is shown in The compressive strength of concrete after different dry–wet cyclic corrosion durations can be calculated by introducing parameters of the damage depth sc is not constant, but is instead a time-dependent process from low to high and gradually to saturation. Costa [sc test data in this paper. The fitting curve is shown in R2 = 0.958. The fitting function is given by Formula (16):When concrete is exposed to an actual chloride environment, the surface chloride content n. Costa  found thD of chloride after different dry and wet cyclic corrosion durations is shown in D of chloride decreased gradually with increasing dry and wet cyclic corrosion durations and had obvious time dependence, i.e., the diffusion coefficient of chloride was not constant. In this paper, the diffusion coefficient D of chloride was modified by the attenuation coefficient n [tD is the actual diffusion coefficient of chloride, t0 is the reference time (t0 = 1 in this paper), and D0 is the chloride diffusion coefficient with a dry and wet cycle inspection period of 1 month.The variation in the diffusion coefficient icient n . The fitThe free chloride content for calculating the damage depth was given above, wherein dx is shown in x(t) = a/2, the corrosion damage zone extends to the whole specimen, and the specimen will crack completely. At this time, the dry and wet cyclic corrosion duration is approximately 253 months (21.1 years), which is much shorter than the 50 years of service life of normal concrete.The prediction of the time-dependent damage depth The free chloride content for calculating the corrosion depth was given above, wherein The time-dependent formula of the compressive strength of corroded concrete in a saline soil area can be obtained by introducing Formulas (18) and (19) into Formula (10):The prediction of the time-dependent compressive strength of corroded concrete in a saline soil environment is shown in The compressive strength of corroded concrete in a saline soil environment first increased and then decreased with increasing dry and wet cycle inspection periods. The deterioration of concrete in a saline soil environment developed gradually from the surface to the interior of the concrete. Therefore, the cross section of the concrete specimens was equivalent to three parts according to the damage depth and corrosion depth: a failure zone, a filling zone, and an undisturbed zone. The compressive strength of concrete was the superposition of these three parts.The compressive strength of corroded concrete in a saline soil environment, as calculated by the mechanical model established in this paper, was in good agreement with the test results, showing a trend of first increasing and then decreasing as the dry and wet cyclic corrosion duration increased. The maximum error was less than five percent.The service life of concrete was predicted to be 253 months (21.1 years) by damage depth and 187 months (15.6 years) by a time-dependent compressive strength model. Both predictions were far less than the normal concrete service life of 50 years. In addition, the long-term compressive strength of corroded concrete was found to be about 90% of that of noncorroded concrete, which did not deteriorate with the corrosion time.The variations in the concrete compressive strength versus the dry and wet cycle inspection periods in a saline soil environment was introduced, and a time-dependent compressive strength model of corroded concrete was established. The main conclusions are as follows:


In a recent article, Balaban and colleagues developed the iTDtest allowing characterization of the type of interactions between different antibiotics at bactericidal concentrations . This visual and semiquantitative assay is designed to determine how antibiotic cocktails affect tolerance and persistence, two phenomena of major importance for the eradication of difficult-to-treat pathogens. Importantly, Balaban and colleagues identified antibiotic combinations allowing for complete clearance of persister and tolerant cells. This commentary discusses the translation of this assay in clinical settings, where antibiotic combination therapies appear to be applied in specific contexts, such as in acute infections or in the case of multidrug or extensively drug-resistant pathogens. The antibiotic resistance crisis is rising worldwide and is announced as one of the next pandemics by the World Health Organization (WHO). By 2050, multidrug-resistant bacteria could cause the death of 10 million people each year. Even more alarming, the WHO warns that the pipeline for new antibiotics discovery and development is drying up with only a few leads under clinical trials.In addition to resistance, antibiotic tolerance and persistence are increasingly recognized as major players in the relapse of infections and emergence of antibiotic resistance . In contEscherichia coli strains. While confirming that ampicillin and rifampicin are strongly antagonist, these researchers bring to light a synergistic effect between ampicillin and kanamycin or streptomycin on the appearance of persister cells. Importantly, adding kanamycin or streptomycin both at high and sub-MICs reduces persistence to ampicillin. Finally, they showed that combining ampicillin, rifampicin and kanamycin completely eradicate persisters formed by an E. coli antibiotic tolerant strain.Classically, the antibiotic tolerance and/or persistence ability of a given strain is evaluated by performing liquid time/kill curve assays . This asMycobacterium tuberculosis infections, or in the case of febrile neutropenic patients (Acinetobacter baumannii, Pseudomonas aeruginosa, Klebsiella pneumoniae, or Escherichia coli strains, antibiotic cocktails have regained interest notably with combination regimens comprising colistin, tigecycline, aminoglycosides, vancomycin, and/or carbapenem, depending on the pathogen (Given that antibacterial drug discovery is a slow process and that the current clinical pipeline is mostly limited to derivatives of classical antibiotic molecules, the short-term approach to combat recalcitrant bacteria might rely on the use of antibiotic combinations , 8. Howepatients . With thpathogen , 11. Antpathogen . Howeverpathogen  and 14.


Developing more resilient and sustainable physical infrastructure increases the demand for sustainable materials and strengthening approaches. Many investigations into strengthening RC beam structures have used either externally bonded (EB) or near-surface mounted (NSM) systems with synthetic fibre reinforced polymer composites. These synthetic fibres are unsustainable since they involve the use of nonrenewable resources and a large amount of energy. Research shows that natural fibre reinforced polymer (NFRP) composites may be an alternative to synthetic FRP composites in the strengthening of concrete beams. However, there is limited literature that validates their performance in various structural applications. Hence, the purpose of this paper is to explore the advances, prospects, and gaps of using EB/NSM NFRP techniques in strengthening concrete beams to provide areas for future research directions. The NSM FRP technique provides improved strengthening effects and mitigates the concerns associated with the EB system, based on a wider range of applications using synthetic FRPs. However, the NSM NFRP strengthening technique has been underutilized, though the EB NFRP system has been more commonly explored in reviewed studies. The knowledge gaps and areas for proposed future research directions are essential in developing work in emerging NFRPs and strengthening techniques for sustainable infrastructure. Concrete today is a very essential component in most construction projects. Its utilization globally is seen to be immense. However, reinforced concrete structures face various levels of deterioration during and after their service life due to various forms of loading conditions and other external agencies. This explains the need for structural enhancement of deteriorating reinforced concrete infrastructure. Several studies have been conducted to evaluate the capacity of natural fibres in improving the flexural, tensile, shear, impact load, and torsional capacity of concrete in order to reduce the demand for synthetic fibres and achieve a more eco-friendly and sustainable construction ,3,4,5,6.Fibres have been engineered over the years to form exterior bonded and near-surface mounted natural fibre composites in reinforced concrete beams, as seen in Several studies have explored the advancements in strengthening concrete structures using FRPs. However, these studies have focused mainly on synthetic fibres such as carbon, glass, basalt, and aramid FRPs, which are the most often utilized reinforcing materials in FRPs due to their high strength and stiffness ,29,30,31An increasing number of people throughout the globe are turning to synthetic fibres such as glass, carbon, or aramid for reinforcement and retrofitting purposes. From considerable research carried out throughout the globe over the past 30 years, we have gained a greater knowledge of the characteristics and behavior of FRPs under various situations . Thus, mIt is true that natural fibres have flaws that must be overcome to compete with glass and carbon. Glass fibres outlast and outperform natural fibres in terms of durability and strength . HoweverTo build a laminate that is effective at reinforcing a concrete beam, it is crucial to comprehend the characteristics, strengths, and limitations of the specific fibre functioning as reinforcement . This knUse of NFRP in flexure applications is gradually gaining significant attention with the current need for a more sustainable means of strengthening deteriorating RC elements, though these studies are limited. Sen & Reddy  created According to Nwankwo & Ede , the useAlso, for RC buildings, Grazide et al.  investigFor enhanced strengthening, some researchers propose merging synthetic fibres with natural fibres to better the mechanical and structural strength of RC constructions because of the downsides  of employing natural fibres . This isIn research by Joyklad et al. , jute fiConsidering a variety of polymer matrices, the FRP thickness and compressive strength of concrete, Yinh et al.  discoverIt is worthy to note that corrosion is a significant contributor to the degradation of concrete ,52. HoweJute fibre reinforced polymer (JFRP) composites were investigated by Jirawattanasomkul et al.  for usagAlam et al.  developeEB Beams with shear-inadequate apertures have yet to be strengthened with NFRP composites through this approach. However, strengthening has been applied using synthetic FRPs as shown by Aksoylu et al. , who conThe demand for safer buildings is becoming more important as the frequency of purposeful and inadvertent explosions continues to rise globally . DraganiEven though torsional stresses can be induced in several structural concrete buildings, design engineers mostly ignored torsion prior to the 1960s. It was anticipated that torsional impacts were small and that the substantial safety factors utilized in flexural design would take care of them. Many incidents of torsional strain and failure have resulted from this notion . The appThe NSM fibre reinforced polymers (FRP) approach is gaining traction as a viable substitute to externally bonded reinforcement (EBR) due to its enhancing of the load bearing capability of RC members. When compared to the EBR approach, the NSM FRP process offers various benefits, including a lower chance of debonding and more durability . HoweverKhalifa  comparedFire occurrences can also be a source of structural deterioration which could be managed using the NSM FRP strengthening technique. This was shown in research conducted by Jadooe et al. , which iIn finding an alternative polymer matrix other than polyester resin or epoxy resin, Al-Saadi et al.  investigIn other ways to improve the efficiency the NSM FRP system in strengthening RC beams, the side near surface mounted system was investigated by Abdallah et al.  through Many studies have been undertaken on single span beam elements, but relatively few on continuous span beams. However, Abdallah et al.  comparedRC beams with pinned and free ends were studied by Capozucca , who exaMinimal research has been documented on the bonding behavior of NSM FRP composites and concrete adopting epoxy glue and evaluated under monotonic loading conditions. Seracino et al.  discoverb is the bar diameter and ab and ba are the smallest and largest widths of the FRP strips shown, respectively.The cohesion failure in a small layer of concrete at the epoxy interface is usually the desired debonding failure mechanism for an NSM FRP-to-concrete contact. If the concrete and CFRP surfaces are correctly prepared and a suitable adhesive is used, then this failure scenario is conceivable to encounter . NSM CFRThere are only a few studies on the shear behavior of RC beams reinforced with the NFRP NSM system as seen on Other investigations on the shear enhancement of RC beams using the NSM system have mostly used CFRP and GFRP as test materials. Studies such as these are important because they provide the groundwork for future research in the field of NFRP. Nonlinear finite element models and practical investigations were carried out by Banjara & Ramanjaneyulu  to invesA wide gap is also seen in the application of the NSM system using NFRP in seismic loading since there little or no studies to validate their applicability and efficiency. However, few studies using CFRP have been conducted to test the effectiveness of NSM system in seismic retrofitting. Esmaeeli & He  in theirReinforcement in polymer composites may be made more environmentally friendly by using natural fibres instead of synthetic fibres. Although natural fibres must be used in greater amounts to give the same strength as synthetic fibres, they are a considerably more cost-effective and rational solution. Reinforced concrete members using NFRP laminates were shown to be more resistant to cyclic stress and to fail at greater displacement values than CFRP laminates. NFRP laminate strengthening of concrete has a bright future thanks to the elimination of most of the objections to the external bonding technique using the near surface mounting method (NSM) . HoweverIt has been discovered that employing natural fibre reinforced polymers (EB or NSM) to strengthen RC beams provides a variety of issues, possibilities, and recommendations, some of which are consistent with earlier studies. On the basis of a thorough evaluation of previous research and the identification of research gaps, the following are the suggested future research areas for strengthening concrete beams in a sustainable and robust manner utilizing natural FRP.Studies concerning the use of NSM natural FRP in strengthening RC beams are scant. This implores the need for further research into such a technique to provide better strengthening to RC beams.Numerical analysis will be required for the validation of experimental findings associated with EB and NSM natural FRP beam strengthening to proffer wider acceptance and applicability.Further studies into the life cycle assessment of the various strengthening techniques using natural FRP will be essential to assess the environmental performance and help improve future strengthening techniques. This is relevant considering the constant changes in the socio-economic and environmental climate.Research into green alternatives to conventional polymer matrices such as epoxy and polyester is encouraged, as they require significant energy input in the manufacturing process and could be toxic, thereby reducing the positive environmental compatibility of natural FRPs.This review also suggests the adoption of innovative technology (such as smart sensors) to give improved insights into the long-term behavior of EB and NSM NFRP reinforced concrete structures.Application of natural FRP in improving torsional resistance of concrete beams is underexplored, hence the need for research is essential as torsional effect is one that can be experienced often in dynamic loading conditions such as strong winds and seismic loads.Research into the application of natural FRPs in the strengthening of beams with unconventional physical dimensions, such as beams with openings, is essential for wider applicability.The need to explore the effectiveness of NFRP in blast resistant beam structures is also essential owing to increasing threats and the need for safer structures as a result of the increasing number of explosions worldwide.There are limitations to research and related issues, particularly with respect to the effectiveness of EB and NSM natural FRP composite systems when coupled loading conditions such as fatigue, cyclic, seismic, impact, and exposure to harsh weather conditions are applied. Research incorporating coupled loading conditions would be valuable to real-life applications of EB and NSM natural FRP strengthening techniques.GFRP and CFRP have been widely used in structural retrofitting based on reviewed studies either using the EB or NSM system. Nonrenewable resources and considerable energy use are required to produce these synthetic fibres, making them unsustainable.Reviewed studies have shown that natural fibre reinforced polymer (NFRP) composites may be an alternative to conventional synthetic FRP composites in structural strengthening.However, their use to validate their performance in various structural applications is underexplored, particularly in their application using the NSM strengthening technique.The NSM strengthening technique using FRPs reduces the flaws associated with the EB technique. These flaws include: delamination, concrete cover separation, and the spread of crushing shear cracks in concrete.Shear and rupture of NFRP are the most frequent mechanism of failure for externally side-wrapped and u-wrapped systems, whereas rupture of the NFRP is the most common cause of failure for fully wrapped systems owing to the confinement effect.NSM synthetic FRPs have shown failure mechanisms through flexural failure by crushing of compressive concrete, flexural failure by rupture, intermediate crack driven pull-out, and end pull-out. However, these structural behaviours remain underexplored using natural FRPs.NFRP strengthening also aids in the transition of a structure from a brittle to a flexible mode of failure via NFRP strengthening.Factors affecting the structural improvement of RC beams employing NSM system include groove sizes and FRP dimensions; clear gaps and edge lengths of NSM FRP; FRP bond length; adhesive material type; and stress restrictions in steel reinforcements. These findings are mainly associated with the use of synthetic FRPs, which are not sustainable.With the development of new techniques that maximize NFRP strength, minimize brittleness and increase ductility, reduce the risk of high temperature exposures and accidental damage, minimize embodied carbon as well as carbon footprints during production, and reduce high capital costs, NFRP adoption, and utilization will grow even more.This review focused on assessing the efficacy of EB and NSM NFRP strengthening procedures in RC beams in terms of structural efficiency empirically. Based on the reviewed studies, the following conclusions can be drawn:


Myoblast differentiation is essential for the formation of skeletal muscle myofibers. Profilin1 (Pfn1) has been identified as an actin-associated protein, and has been shown to be critically important to cellular function. Our previous study found that PFN1 may inhibit the differentiation of bovine skeletal muscle satellite cells, but the underlying mechanism is not known. Here, we confirmed that PFN1 negatively regulated the myogenic differentiation of bovine skeletal muscle satellite cells. Immunoprecipitation assay combined with mass spectrometry showed that Cdc42 was a binding protein of PFN1. Cdc42 could be activated by PFN1 and could inhibit the myogenic differentiation like PFN1. Mechanistically, activated Cdc42 increased the phosphorylation level of p2l-activated kinase (PAK), which further activated the phosphorylation activity of c-Jun N-terminal kinase (JNK), whereas PAK and JNK are inhibitors of myogenic differentiation. Taken together, our results reveal that PFN1 is a repressor of bovine myogenic differentiation, and provide the regulatory mechanism. Satellite cells are a type of specific adult stem cell population of skeletal muscle, which are normally quiescent. However, satellite cells can be strongly activated after tissue injury, and begin to divide and proliferate into myoblasts. These myoblasts then fuse and differentiate into muscle fibers, thus supporting skeletal muscle regeneration . MyoblasPTEN gene, thus inhibiting the AKT pathway, resulting in the increase of p27 protein [PFN1 is one of the important actin-binding proteins, the molecular weight of which is about 15 KDa. The tertiary structure of PFN1 is mainly composed of four α helixes and seven β folds . Mutatio protein . The bin protein . Another protein . Some st protein ,11,12. W protein . TherefoThe small GTPase protein, Cdc42, is a member of Rho GTPase and is widely present in cells. Cdc42 can regulate many physiological processes, such as cell proliferation, movement, polarity, cell division, and growth . In mousIn this study, we find that PFN1 negatively regulates the differentiation of bovine skeletal muscle cells. Cdc42 is identified as a binding protein of PFN1, and has a negative regulatory effect on the differentiation of skeletal muscle cells consistent with PFN1. Mechanistically, PFN1 interacts with Cdc42 to stimulate Cdc42 activity, thereby activating PAK and JNK signaling pathways to inhibit bovine myoblastic differentiation.2 incubator until the cell density reached 80%, and the proliferating cells were collected, or the differentiation medium containing DMEM with 2% horse serum  and 1% penicillin-streptomycin liquid was aspirated to induce the cells to differentiate. After culturing in cell differentiation medium for 2 days (DM2), experiments were usually carried out.The bovine myoblasts used in this study were prepared in our lab; the isolation, identification, and induced differentiation procedure were described in our previous study . Myoblas® Cdc42 Activation Assay Biochem Kit™  was used.The primary antibodies used in the study included rabbit anti-Profilin1 , rabbit anti-Cdc42 , rabbit anti-PAK , rabbit anti-Phospho PAK , rabbit anti-IgG , rabbit anti-JNK , rabbit anti-Phospho JNK, mouse anti-MyHC , mouse anti-MyoG , mouse anti-GAPDH , and mouse anti-tubulin . The secondary antibodies included horseradish peroxidase goat anti-mouse/rabbit IgG , PAK inhibitors , and JNK inhibitors . A G-LISA® 96 Instrument . GAPDH was used as an internal reference gene. The relative expression level of genes was calculated by the 2−ΔΔCt method. The sequences of all primers used were listed as follows: GAPDH forward: 5′-TGTTGTGGATCTGACCTGCC-3′, reverse: 5′-AAGTCGCAGGAGACAACCTG-3′; MyoG forward: 5′-GGCTGACAAATGCCAGACTATCC-3′, reverse: 5′-TGGTCCCTTGCTTTATCTCCCT-3′; MyHC forward: 5′-CTGGAATCCGGAGGCAGAA-3′, reverse: 5′-TTTTCGAAGGTAGGGAGCGG-3′; Cdc42 forward: 5′-AAACAGCTGCCCCTACTGTC-3′, reverse: 5′-GGTTTGTTAGGGCTGCCTGA-3′; PFN1 forward: 5′-GTGGAGCCCCAACCTTCAAT-3′, reverse: 5′-TTGATCATACCGCCGTGGAC-3′.The total RNA of cells were isolated using the Cell RNA Extraction Kit . Then, the first strand cDNA was prepared using PrimeScript II 1st Strand cDNA Synthesis Kit . qRT-PCR was performed using All-in-One™ qRT-PCR Mix  in a LightCyclerBovine myoblasts were washed by PBS buffer and lysed in RIPA buffer  supplemented with PMSF for total protein characterization. Then, equal amounts of cells lysate were resolved by 10% or SDS-PAGE and transferred onto PVDF membranes . The membranes were blocked with 5% BSA for 1h, incubated with primary antibody at 4 °C for 2 h or overnight, then incubated with secondary antibody for 1h to detection using ECL chemiluminescent substrate (Solarbio). Quantitative analysis was carried out by calculating the gray value, which was calculated using ImageJ software  and the value of the target protein was normalized to internal reference protein.PFN1 and Cdc42 to the pcDNA3.1(+) vector, respectively. The pcDNA3.1(+) was a negative control. All the primer sequences of gene cloning are listed in PFN1 and Cdc42 knockdown and negative control sequences (si-NC) were synthesized by RiboBio ; the siRNA sequences are listed in We obtained the recombinant plasmids pcDNA-PFN1 and pcDNA-Cdc42 by ligating the CDS regions of Co-IP was performed using a Beaver BeadsTM Protein A/G Immunoprecipitation Kit . Briefly, the protein of DM2 myoblasts were collected using binding buffer supplemented with 1× protease inhibitor cocktail. The pretreated magnetic beads protein A/G and the PFN1 antibody or IgG antibody were incubated for 30 min at room temperature. Then, the beads–antibody complex and cell lysates were incubated with rotating at 4 °C overnight. Finally, the supernatants were subjected to SDS-PAGE and mass spectrometry (MS) identification by Wuhan GeneCreate Biological Engineering Co., Ltd .® Cdc42 Activity Assay Biochem Kit . Briefly, the cells were lysed with lysis buffer containing 1% protease inhibitor cocktail, and the diluted Cdc42 primary antibody was added and incubated for 15 min in the micro-vibrator. Then, the diluted Cdc42 secondary antibody was added and incubated for 30 min in a micro-vibrator. Finally, the mixed HRP A/B was added, incubated at 37 °C for 15 min, and the absorbance value of 490 nm was immediately measured.A Cdc42 activity assay was performed using a G-LISAp < 0.05 was considered statistically significant; * p < 0.05, ** p < 0.01, and *** p < 0.001.All of the experiments were performed at least three times. All results are presented as the mean ± SEM. Statistical analyses of differences between groups were performed using a two-tailed Student’s t-test, and To examine the expression of PFN1 during myoblast differentiation, the mRNA and protein expression levels of PFN1 at different differentiation days were assessed. The results showed that the expression of PFN1 decreased significantly during the myoblast differentiation. To determine the function of PFN1 in this process, we transferred PFN1 siRNA or pcDNA-PFN1 into the bovine myoblasts to gain the knockdown or overexpress of PFN1 cells, respectively. Myoblast differentiation marker genes, MyHC and MyoG, were detected by qRT-PCR and Western blot in DM2. We found a significant increase both at the mRNA and protein levels of MyHC and MyoG in PFN1 knockdown cells, whereas the mRNA and protein levels of MyHC and MyoG significantly decreased in PFN1 overexpression cells A–D. MoreAccording to previous research, PFN1 usually plays a regulatory role by interacting with other proteins. Thus, we attempted to identify the interacting protein of PFN1 to analyze the regulation mechanism of PFN1 on myogenic differentiation. We successfully obtained the binding protein of PFN1 in DM2 myoblasts by immunoprecipitation technology . Then, tThese pathways are also related to muscle development, which further proves the key regulatory role of PFN1 in myogenic differentiation. With the aid of the previous bioinformatics analysis, it was found that the differential binding proteins were mostly related to actin and small GTPase, and these proteins were also related to muscle development. Protein–protein interaction (PPI) analysis of PBPs by the STRING database revealed that PFN1 interacted with Cdc42, RhoA, RhoC, VASP, and so on F. After Further, in order to validate the binding of PFN1 and Cdc42, proteins of DM2 myoblasts were collected and subjected to CoIP. The IP efficiency was detected by Western blot. The results showed that Cdc42 was significantly enriched in the PFN1-IP group compared with negative IgG G. Thus, Next, to illuminate the mechanism of PFN1 cooperating with Cdc42 in regulating the cell differentiation, the effects of PFN1 on Cdc42 were measured both at mRNA and protein levels. The results showed that changing the expression of PFN1 did not significantly effect the mRNA level of Cdc42 in DM2 A; howeveConsidering that although it had been reported that Cdc42 could regulate myoblast differentiation in other species , the effBased on the above findings, we concluded that PFN1 activated Cdc42 activity by binding to Cdc42, consequently inhibiting the cell differentiation.Based on above data, we speculated that PFN1 could negatively regulate the differentiation of bovine myoblasts by activating Cdc42. Uncovering the detailed regulation mechanism of PFN1-Cdc42 on inhibiting myogenic differentiation is needed. Some studies have reported that Cdc42 can activate the PAK signaling pathway to affect cell activity, and the PAK signaling pathway has been confirmed to play important roles in the differentiation of myoblasts. Thus, we verified whether Cdc42 regulates myogenic differentiation through the PAK signaling pathway in bovine myoblasts. The phosphorylation of PAK is a sign of activation of the PAK signaling pathway. We found that the knockdown of Cdc42 could decrease the phosphorylation levels of PAK in DM2 of the bovine myoblasts A, whereaCombining all the results, we considered that PFN1-Cdc42 could activate the PAK/JNK signaling pathway, thereby negatively regulating the differentiation of bovine myoblasts.PFN1 is an evolutionarily conserved small actin-binding protein that regulates many cellular activities, including proliferation and motility . AccordiAn increasing amount of research is showing that PFN1 can act as a key signaling molecule in the network hub, and participate in a series of important physiological processes, such as protein transmembrane transport, nuclear transcription, and activation of the small GTPase signaling pathway, by binding to target proteins through its domain . Thus, wAs a Rho GTPase, Cdc42 plays a key role in a variety of cellular processes, and activated Cdc42 can mediate signaling cascades and activate various downstream effectors, thereby leading to many cellular activities, such as cell adhesion, motility, polarity, cytokinesis, and growth ,27,28. IActive Rho GTPases can interact with a number of downstream effectors, of which more than 200 effectors have been identified, e.g., some protein kinases such as ROCK and PAK family members, and scaffold proteins such as mDia1 and WASP . PAK hasIn conclusion, we confirmed that PFN1 played an important negative role in the differentiation of bovine myoblasts. Cdc42 is identified as a binding protein of PFN1, and has a negative regulatory effect on the differentiation of myoblasts consistent with PFN1. Mechanistically, PFN1 interacts with Cdc42 to stimulate Cdc42 activity, thereby activating PAK and JNK signaling pathways to inhibit bovine myoblastic differentiation.