Copper slag (CPS) is a large amount of waste material produced during the manufacture of copper. The disposal of this waste material becomes a problem for environmental concerns. Therefore, it is necessary to explore feasible alternate disposal options. They may also be utilized in concrete manufacturing to cut down on the usage of cement and natural aggregates. A lot of researchers focus on utilizing CPS in concrete, either as a cement replacement or as a filler material. This article aims to summarize the literature already carried out on CPS in conventional concrete to identify the influence of CPS on the fresh, hardened and durability performance of cement concrete. Results indicate that CPS improved the strength and durability performance of concrete but simultaneously decreased the slump value of concrete. Furthermore, an increase in the durability performance of concrete was also observed with CPS. However, the higher dose results declined in mechanical and durability aspects owing to a scarcity of flowability. Therefore, it is suggested to use the optimum dose of CPS. However, a different researcher recommends a different optimum dose ranging from 50 to 60% by weight of fine aggregate depending on the source of CPS. The review also recommends future researcher guidelines on CPS in concrete. A widely utilized raw material in construction is concrete, which is the basis for all construction and development initiatives around the globe, serving as the base for all buildings and infrastructure ,2,3. The2 [2 emissions, waste materials should be used instead of cement in concrete.The term “sustainable building” refers to management that is accountable for providing a favorable environment that considers ecological and resource development ,11,12,132 ,18,19. C2 . The amo2 ,21. Desp2 . To miniThe industrial sector has seen significant expansion, resulting in a vast number of by-products whose dumping has come to be a serious problem, since it impacts the ecosystem’s ,24,25. TCopper slag (CPS) is a metallurgical waste product that is created through the matte smelting of copper metal. CPS is an industrial waste substance that is formed as a by-product of the copper production process. It is a smooth and glassy by-product of the matte smelting and refining phases involved in the pyrometallurgical removal of copper. Copper is extracted and purified from copper oxide ores using aqueous (water-based) solutions at room temperature, often in three steps: heap leaching, solvent extraction and electrowinning.Heap leaching is the method of extracting metals from chemical solutions by allowing them to percolate. Low-grade ore that would otherwise not be economically sent through a milling process is often utilized in heap leaching. The crushed ore is placed into a heap on top of an impermeable layer, on a little slope, after mining, shipping and crushing to a constant gravel or golf ball size. The copper from the ore is dispersed in the leaching agent (diluted sulfuric acid), which is sprayed via sprinklers on top of the heap pile and allowed to flow down into the heap. A small pool is used to collect the copper sulfate and sulfuric acid “pregnant” leach solution that results. Currently, concentrations of the copper complex range from 60 to 70 percent.The second stage is solvent extraction, which involves stirring and allowing two immiscible (non-mixing) liquids to separate, causing the copper to transfer from one liquid to the other. A solvent is aggressively combined with the pregnant leach solution. The copper migrates into the solvent from the leach solution. The two liquids are then allowed to separate depending on solubility, with the contaminants staying in the leach solution while copper remain in solution in the solvent. The remaining leach solution is then recycled by adding more acid and returning it to the heap leaching sprinklers.The last stage is an electrolysis process known as electrowinning. An inert anode (positive electrode) and the copper solution from the prior phase, which functions as an electrolyte, are both contacted by an electrical current. Next, 99.99 percent pure copper is deposited onto a cathode (negative electrode) as positively charged copper ions (referred to as cations) emerge from solution. The manufacture process of copper slag in the industry is displayed in It has been calculated that the copper industry generates about 24.6 million tons of slag per year across the globe. Even though CPS is frequently employed in the sandblasting business and the manufacture of abrasive tools, the rest is placed without any further usage or recovery possible. CPS contains mechanical and chemical properties that allow it to be utilized in concrete as a substitute for cement or sand, depending on the application. For example, copper slag possesses a variety of mechanical features that make it a desirable choice for aggregate application, including great soundness characteristics, outstanding abrasion resistance and excellent stability . BecauseFollowing the escalation of the issue, various investigations have shown a wide range of potential reuse and recycling options for this particular material. Among the potential options for metal recovery from slags containing significant levels of metallic elements are different techniques such as electric arc furnace melting, leaching and flotation (as well as other methods). However, since the metals present in copper slag are often found in trace levels, their recovery may not be economically feasible in most cases. Instead, various applications for copper slag were investigated and eventually accepted . An alte2O3, SiO2, and Fe2O3. Pozzolans are classified as siliceous or siliceous and aluminous minerals that have no or little cementitious properties; however, when finely ground, they chemically react with calcium hydroxide (CH) in the presence of water to produce compounds with cementitious characteristics . It is possible to reduce disposal costs by using CPS in concrete as binder or filler. This may also assist with conserving the environment by lowering the amount of waste produced. Because of the massive amount of CPS being generated, environmental pollution has an adverse effect on the growth of the nation. The effective and environmentally friendly use of CPS, as well as the promotion of green building, are significant concerns of this assessment.Additionally, copper slag has pozzolanic capabilities due to its low CaO level, as well as the presence of other oxides such as AlAlthough some studies have discovered that copper slag possesses pozzolanic activity, the activity is quite low, limiting the use of copper slag as a mineral additive in concrete ,37.2, Al2O3 and Fe2O3) in the clinker) of the activator increases the hydration degree of quick-cooled copper slag and the polymerization of amorphous hydration products. In quickly cooled copper slag, instead of high Fe particles, high Si particles provide primary activities. Alkali-activated quick cooled copper slag mortars exhibit reasonably high compressive strengths, particularly when the silica modulus is greater than one, indicating that alkali-activated copper slag for building may be feasible in the future [The silica modulus  of CPS is shown in 2, MgO and Fe2O3 as the primary constituents [2, Al2O3, CaO and MgO, which account for 95 percent of the total composition. It also contains manganese, iron and sulfur compounds, as well as tiny quantities of numerous other elements [Four to five percent alumina, four to six percent calcium oxide, thirty-five percent to thirty-seven percent iron, thirty percent to thirty-four percent silica, and one percent copper are the primary elements of CPS . Other stituents . Iron slelements . Accordi2O6), fayalite S(iO4Fe2), anorthite (CaAl2Si2O8), quartz (SiO2) and magnetite (Fe3O4) [3O4). The amorphous nature of the SiO2 found in CPS has a significant impact on concrete, from the initial hydration to the ultimate strength [X-ray diffraction (XRD) analysis revealed that the mineralogical components included in this slag are pyroxene (CaZnSi (Fe3O4) . Figure strength . Accordistrength , it is pw/c). Concrete made with w/c 0.55 possess high flowability (100–175 mm) to have a maximum slump value of 175 mm, and therefore higher workable concrete. However, the slumps of copper slag mixes were less than control concrete for a 0.45 w/c and high percentages of copper slag are substituted. In such a case, concrete has very little workability, which could be unsatisfactory for several useful purposes. Furthermore, lower workability also adversely affects strength properties.It was hypothesized that the CPS would experience more slump collapse (more workable concrete) when compared with natural aggregate due to lower water absorption. However, the findings proved that the supposition is wrong. Slumps were probably influenced to some degree due to the angular shape of CPS, which enhances the friction between concrete components and ultimately reduced the flowability of concrete. In addition, a researcher discovered that when sand was substituted with CPS, the workability of the concrete decreased, contrary to what was expected . WorkablAccording to one investigation, the recorded slump was 150 mm when CPS was utilized to substitute 100 percent of the copper. According to the authors, the low water absorption features of CPS and its glassy surface when compared to aggregate caused in a noteworthy increase in the flowability of concrete due to a significant rise in the amount of free water remaining after the absorption and hydration processes were completed. The improvement in flowability of concrete may be made with the same amount of sand substituted, and these mixes may have better flowability, as well as higher strength and durability than standard high-performance concrete (HPC) mixes. Furthermore, it was highlighted that mixes containing significant levels of CPS exhibit bleeding and segregation which might have a harmful influence on the strength of concrete .2 decreases, while small particles of mineral mixture fill the spaces between cement particles, making the cement mortar denser and strengthening the interfacial area [The compressive strength (CMS) outcomes at each curing interval are shown in ial area . Accordiial area . Under sial area . The majial area ,67. HoweNumerous studies have indicated that the CMS and TS of concrete specimens created using CPS as fine and coarse particles are higher than those of regular concrete . As a co2 value greater than 90%. The flexural strength (FS) of concrete for all mixtures at each curing time is shown in Analysis of FS of the mortar containing 10 percent CPS reveals that FS increased by 12.9 percent more than the control mixture. According to some theories, this improvement may be attributed to the increase in the compactness and durability matrix of the CPS mortar. Despite this, the findings of the 20 and 30 percent CPS tests indicate that the gain in FS is much more significant. The improvement in FS of the 20 percent CS mortar was about 18.2 percent higher than the FS of the control mortar. The concrete containing 30 percent CPS resulted in the greatest improvement in FS, which was 38.7 percent more than the control mortar. This substantial increase in FS was most likely due to the increased compaction and uniformity of the distribution of CPS in mortar mixes, which has resulted in a successful improvement in FS . The FS FS is almost 10% to 20% of CMS varying on the mix design of concrete. It can be noticed that the trendline among CMS and FS seems to be straight. Therefore, a strong correlation has occurred among CMS and FS with an R2 value more than 90%. w/c = 0.55, higher water absorption was exhibited compared with blank concrete. The w/c = 0.45 mixes showed less water absorption when compared to the corresponding control mix, except for the two mixes with the greatest CPS proportions, i.e., 80 and 100% which had the maximum water absorption. However, all the mixes had low water absorption rates of less than three percent. This was within predicted limits for high durability in terms of liquid infiltration in concrete, which was observed by the researcher [w/c = 0.55 mixes, was caused by an excess of water, i.e., CPS grains showed less water absorption than sand, which might have resulted in a larger porosity of the mixture [The average water absorption of the mix with CPS substation is shown in searcher . The ris mixture . Accordi mixture . Similar mixture . Water a mixture .The percentage of voids in concrete mixes containing CPS is shown in w/c ratios of 0.45 and 0.55, respectively) showed more corrosion (as seen by the larger mass loss of the bars) than the concrete made with CPS. The results of the faster corrosion assessment in terms of mass failure of the entrenched reinforcement are depicted in w/c = 0.55 mix with 100 percent CPS demonstrated only a minor loss in mass compared to the other mixtures. Utilizing CPS as a substitute for sand, the researchers discovered that the corrosion resistance of the resultant mix was greater than that of mix made with natural sand. However, reverse results were examined by Brindha et al. [The 0.45 control mix had the greatest amount of corrosion, as well as several surface fractures near the reinforcing bar, which were the most severe. However, the a et al. , who demThe concrete including various amounts of CPS percentages was preserved in water for 28 days before being subjected to the sulfuric acid mixture for 56 days and examined for corrosion. The compressive capacity and mass of concrete sample were measured before and after contact with acid to determine the severity of the acid assault on the specimens. When exposed to a sulfuric acid solution for 56 days, the mass of concrete specimens varied, as shown in The electrical resistance is defined as the voltage age ratio multiplied by the electrical current that runs across a sample (voltage to current ratio). It is also referred to as material resistance to electric current in certain circles. Understanding the strength of the flow of the electrical current is important, since it may aid in predicting the probability of reinforcing corrosion occurring. The fluctuation in electrical resistance with hardening time and the percentage of CPS is shown in Extremely serious environmental deteriorations brought on by sulfate assault have an impact on the long-term durability of concrete buildings. Structures made of concrete, such as foundations, bridges, piers, concrete pipelines, etc., experience expansion and cracking as a result of sulfate assault, which worsens the condition. If the sulfate ions originate from sea water, ground water or soil, they will be present in the solution together with other ions including magnesium, sodium, calcium and potassium . The finIn another investigation, iron slag was used as a replacement for fine aggregates in the SCC, and it was shown that specimens attacked by sulfate showed no mass loss despite developing white deposits after 91 days. After 28 days of sulfate exposure, it was discovered that each degree of iron slag replacement had a lower-than-5% effect on the compressive strength . Sharma 3), larnite (C2S), and ettringite are the major minerals in the paste samples [X-ray diffraction (XRD) patterns after 7, 28 and 90 days of curing and the blended cement pastes containing 30 percent copper slag are shown in  samples .2S is present as a non-hydrated cement component [Portlandite is produced when cement hydrates, and it is the main crystalline mineral in the pastes. A study also claimed that CH forms weak pockets which adversely affect the strength of concrete . Furtheromponent . In addiIt is generally known that the reactive element in pozzolanic materials, amorphous silica, may interact with CH to create more hydration products. The consumption of CH in cement pastes is often used to measure the intensity of pozzolanic reaction. In mixed cement pastes, the CH diffraction peaks are located at 18.02 and 34.05 degree. As indicated in 3 into account to precisely quantify the level of pozzolanic reaction [The findings indicate that this peak intensity gradually and noticeably decreased throughout the course of the curing process, with a residual amount still present in the pastes after 90 days. This suggests that the pozzolanic reaction results from the inclusion of UGCS and would continue even after 90 days of cure time. In the part that follows, a quantitative study will be carried out while taking the impact of CaCOreaction .Copper slags are used in the production of cement, aggregates, landfill, ballast, abrasives, roofing granules, glass, tiles and bituminous pavements, among other things. Copper slag has lately been studied in terms of its properties and applications . The bigHowever, The Cu and Pb concentrations exceed the authorized limits  at the 30 percent replacement level of CPS, whereas the other elements  are within the limits. Cr has the lowest concentration (it is close to the detection limit of 0.003 mg/L). When the CPS content is raised to 50 wt.%, the excess quantities of Cu and Pb concentrations rise as well compared with the 30 wt.% content instances. Excess concentrations of the metals Zn, Ni and As are also found. The findings clearly reveal that employing CPS (both air cooled and water cooled) as a partial cement replacement offers a significant danger of heavy metal leaching into the environment , particularly at high replacement levels. Other CPS applications, such as aggregates in concrete and raw materials for alkali activation, may also provide a risk of heavy metal leaching .The results of the leaching procedure (TCLP), acid leaching and repeated extraction tests performed on a large number of slag samples of varied compositions produced from the use of numerous copper concentrate show that heavy metals have little leachability and that long-term stability is assured even in harsh environments. Leaching experiments on mechanically activated samples provide an indication of the heavy metals’ resistance to leaching even after weathering. The heavy metals contained in the slag are stable, according to numerous extractions leaching experiments, and are unlikely to dissolve considerably even after repeated leaching in an acid rain environment. The greatest concentration of all components is significantly below the USEPA 40CFR Part 261 permitted limits . A studyIt is indicated that the slag is safe for use in a broad range of applications, including Portland cement, building materials such as tiles and bituminous pavement projects. The slag samples are non-toxic and pose no risk to the environment.The focus on green construction has resulted in an ongoing search for alternative materials to be employed in concrete construction. In this review, a complete parametric analysis was carried out to determine the impacts of CPS on physical and chemical properties, concrete qualities both fresh and hardened and concrete’s long-term durability performance. Consequently, the findings are given below.The physical property of CPS shows that the particle nature of CPS is rough and angular which adversely affects the flowability of concrete.The chemical composition of CPS ensures that it can be used as binding material.The slump value of concrete was reduced with the replacement of CPS due to angular and rough surface texture.The setting time increased with CPS as the pozzolanic reaction proceeded slowly.CPS up to 60% can be used without any harmful impact on the mechanical strength of concrete. The improvement in compressive, split tensile strength and flexure at 28 days was 9%, 6% and 9% higher than control concrete, respectively.The higher dose of CPS (80 and 90%) resulted in a decline in the mechanical strength of concrete due to the absence of flowability.2 value greater than 90%.A good correlation was observed between two specified strengths with an RThe durability performance of concrete, such as water absorption and voids, corrosion resistance, acid resistance and electric resistivity increased with CPS.SEM results reveal that the performance of concrete with CPS can be improved with the addition of secondary cementitious materials.The overall studies demonstrate that the CPS has the credibility to be utilized partially in concrete, either as a binding material or as a sand. The optimal percentages are an essential parameter for good strength. Different researchers recommend a different optimum value of CPS due to a change of source. The typical range of optimum value of CPS is from 50 to 60% by weight of fine aggregate. Furthermore, less information is available on dry shrinkage and creep properties of concrete with CPS. No or little information is available about the alkali silica reaction (ASR) connected with the CPS substitution. Lastly, although CPS can be utilized in concrete and the mechanical capacity can be enhanced, concrete is still low in tension. Therefore, further research was suggested to enhance the ductility of concrete with the supplement of various kinds of fibers. A study  conclude


Understanding pregnant women and mothers’ perceptions towards antibiotic use and resistance is essential for appropriate antibiotic use and limiting antibiotic resistance. This study aimed to explore perceptions and reported practices of pregnant women and mothers with children under two years of age regarding correct antibiotic use and antibiotic resistance in Vientiane Province, Lao PDR.The study employed an exploratory qualitative research design using focus groups discussions (FGDs). Participants were purposively selected based on: being pregnant at third trimester and attending antenatal care and mothers with children under two years of age, attending the health facility for postpartum visit /vaccinations. Six focus group discussions were conducted in September 2019 with a total of 55 women. The FGDs were transcribed verbatim, data were analyzed first by coding then categorizing the data as we looked for patterns and themes by using the qualitative content analysis.Most participants had some understanding of antibiotics but wrongly believed antibiotics can be used to treat viral disease. Over half of the participants had heard the term “antibiotic resistance”, but often believed it was their bodies, not the bacteria that developed antibiotic resistance. During pregnancy and for their infants, women preferred to use antibiotics only when prescribed by a doctor. Outside of pregnancy however, consuming antibiotics without a prescription was commonly reported. Participants wanted more information about the indications for antibiotic use and antibiotic resistance.More effort is required to increase the level of understanding, and practice of mothers to promote optimal antibiotic use. Mothers’ desire to learn more, and their fundamental concern for their children, can be used to promote appropriate antibiotic use. Awareness raising should be complemented by efforts to address other determinants of inappropriate antibiotic use, including educating healthcare workers, and pharmacists and addressing health service determinants that contribute to inappropriate antibiotic use. Pregnancy is a period of great physiological changes with pregnant women typically more susceptible to bacterial infections than non-pregnant women , 2. CommInappropriate use of antibiotics during pregnancy and in children under two years of age, is also considered an important contributor to antimicrobial resistance, an issue of global concern , 15. InaIn low and lower middle-income countries, inappropriate antibiotic use is also often facilitated by over the counter availability of antibiotics , 23–27. The Lao People’s Democratic Republic (Lao PDR) is a lower middle-income country situated in South East Asia. Information about understanding of antibiotic use and resistance is scant, but there is evidence of inappropriate antibiotic use. Studies suggest that purchasing antibiotics over the counter without a prescription is common , 32–34. Inappropriate antibiotic use during pregnancy and infancy may put the mother and child at greater risk for maternal and child adverse outcomes as well as increased antibiotic resistance. Studies on perceptions and reported practices of pregnant women and mothers of children under two regarding antibiotic use and resistance is lacking in Lao PDR. Therefore, it is critical to understand pregnant women and mothers’ perceptions towards antibiotic use and resistance in order to improve appropriate antibiotic use, especially among these vulnerable groups.The purpose of this study was to explore perceptions and reported practices of pregnant women and mothers of children under two regarding antibiotic use and resistance in Lao PDR. This study could provide useful information for future intervention aiming to improve appropriate antibiotic use contributing to contain antimicrobial resistance.An exploratory qualitative research design using focus groups discussions (FGDs) was chosen to obtain the rich, in-depth information needed to understand and interpret knowledge, experiences and practices of pregnant women and mothers having children two years of age. Group discussions can generate in-depth understanding of participants’ experiences and beliefs and allow for an exploration of multiple dimensions of an issue under examination , 37 ThisThe study setting was Thoulakhom district in Vientiane Province. Vientiane Province has a population of 419,000 inhabitants and has 11 districts. Thoulakhom district, with a population of 57,791 inhabitants , was purParticipants were purposively selected based on being (i) pregnant at third trimester attending antenatal care (ANC), and (ii) mother with children under two years of age, attending the health facility. Pregnant women were identified via the local health ANC registry and invited to participate during their ANC visit if they gave informed consent. Mothers who participated in FGDs were identified during a postpartum visit and/or child vaccination visit and approached by local health staff who informed potential participants of the study and its objectives. All health facilities including district hospital and health centers in Thoulakhom district were selected using simple random sampling based on the health facility name list available at the district health office.At each selected health care facility, two FGDs were conducted, one with pregnant women and one with mothers with children under two years of age. Three groups of pregnant women in the third trimester with a total of 29 participants and three groups of mothers with children under two years of age with a total of 26 participants were conducted. Each FGD consisted of eight to eleven participants with varying occupations, age, education, number of pregnancy and children background. Altogether, six FGDs with a total of 55 participants were included. A semi-structured interview guide for FGDs was developed and pretested among one group of pregnant women at third trimester with eight participants and one group of mothers of children under two with eight participants at Phonehong district (Vientiane province), with similar characteristics as the target population. Participants were asked about their perceptions towards antibiotic side-effects and resistance, consequences of antibiotic resistance, experience with antibiotic use, information seeking and preferred sources of information. Based on the feedback from the pretest, the interview guide was adjusted and finalized.The FGDs were guided by three of the researchers . The FGDs were conducted at the meeting room of the district hospital or health centers. Each FGD lasted 60–90 min and was closed when no new ideas were emerging. Participants were coded according to parity status (pregnant women and mothers with children under two years of age). Each participant received a small gift of towel to thank them for their time.The audio records and notes of the discussions were transcribed verbatim. The Lao transcripts were translated into English and manually analyzed using manifest and latent qualitative content analysis , 39. We The validity of the analysis was ensured by: (1) the triangulation of information; (2) coding undertaken by three researchers  independently, with differences discussed between coders to reach agreement. In reporting the findings, codes are used to maintain anonymity of the key informants.Ethics approval was obtained from the National Ethical Committee for Health Research, Lao PDR . All methods were carried out in accordance with relevant guidelines and regulations. In addition, prior to undertaking the study, the study design and purpose were discussed with the head of each community health center and their approval gained. Participants and the researchers did not know each other, but participants were aware of the interviewers’ status as researchers. Verbal informed consent was gained from each participant after an explanation of the study objectives, assurance of confidentiality and that choosing not to participate would not disadvantage them in any way.Overall, 29 pregnant women and 26 mothers of children under 2 years of age participated in six FGDs. The mean age of the pregnant women was 27 years (range 17–40), most were housewives and rice farmers, slightly more than half (17/29) had completed primary education (Table Most participants were aware that antibiotics can have side-effects with anaphylaxis as the most commonly cited. Some recounted experiences of breathing difficulties after taking Ya-am-pi (ampicillin) or skin rashes:“I took antibiotics, and I had some adverse event such as feeling sleepy. I was allergic to ampicillin injection which caused respiratory distress and dyspnea” .One pregnant woman thought taking antibiotics during pregnancy may lead to congenital malformation. Other negative side-effects reported by participants included feeling weak, fatigue, vertigo and weight loss in pregnant women. A few participants thought that taking too many antibiotics could dry the womb out, resulting in miscarriage and infertility:If the pregnant women have overconsumption of antibiotics, their pregnancy could have been aborted. The older people were told not to take too many antibiotics, but we don’t know how bad it is. I thought that it could affect t the bone.” .““The side effect of antibiotic was that if women take too much antibiotics in a situation where it is not needed, the uterus can be dry, they could be thin, making them less likely to get pregnant or infertile” .Another reported side-effect was fatigue. Some mothers felt taking antibiotics after delivery could decrease breast milk production and cause a dry throat.“I thought that the adverse event was like the side effects of antibiotics overconsumption where it would make you thin, dry throat and no breast milk” . Over half of the participants had heard the term “antibiotic resistance” or in Lao language as “ Kan tan to ya tan xeua” which they described as a condition whereby infections could not be cured by previously used antibiotics, requiring a change of antibiotics. Typically, women understood it was their bodies, not bacteria, that developed antibiotic resistance. Understanding of antibiotic resistance was also informed by the women’s’ experiences as illustrated below:Antibiotic resistance is taking antibiotics without a prescription from the doctor in which the medicine works effectively at the beginning but fails the second time. My son often had pneumonia and went for check-up. The doctor prescribed antibiotics, but it was not effective. Later, the doctor tried to prescribe a different antibiotic and my son recovered.” .“Some mothers cited a link between antibiotic resistance and superbugs. Examples women gave of antibiotic misuse included substituting the prescribed antibiotic for a different one, inappropriate use of antibiotics, taking antibiotics for only mild symptoms or self-medicating. The following quotes highlight some of the mother’s understanding of how antibiotic resistance is caused:“Whenever people got sick and take an antibiotic by themselves without doctor’s prescription, they will become antibiotic resistance”  “Antibiotic resistance is the overconsumption of the antibiotic, but it is not effective anymore and they need to change to a new type of antibiotics. If the patients follow the doctor’s advice and completed the antibiotic treatment, there will not be a medicine resistance” .Not all participants understood the meaning of antibiotic resistance however, illustrated in the quote below:“I had no ideas regarding the antibiotic resistance. Our family did not usually use the antibiotics unless we have some wounds” .“Resistance to the antibiotic had impact to the human body  leading to severe complications and some fatalities. However, the consequences of antibiotic resistance were the physical deterioration which can be life threatening” Most participants recognized antibiotic resistance could harm human health, as one participant explained:“The consequences of antibiotic resistance were whenever we had pharyngitis, pneumonia, the patients took antibiotic orally, which could not be cured, so they will change to antibiotic injection. The consequences of antibiotic resistance will keep the infections reoccurring” Most participants understood antibiotic resistance makes it harder to treat some infectious diseases and felt it could lead to adverse effects, such as prolonged fever, weakness, loss of appetite, and increasing severity of the condition being treated. One pregnant woman explained that antibiotic resistance could lead to recurrent infections:“Some patients took amoxicillin or ampicillin wrongly which they could have an allergic, and trouble breathing, and I think that this is the consequence of antibiotic resistance” One mother linked antibiotic resistance to potential side-effects such as rashes and respiratory distress:Most participants said that they preferred not to take antibiotics while being pregnant. When not being pregnant however, several reported buying antibiotics from private pharmacies to treat the common cold, pharyngitis, or fever. Commonly used antibiotics were amoxicillin or ampicillin. Some participants said they had taken antibiotics for a headache, based on blood test results which showed increased level of white blood cells, dysuria, and leucorrhea, skin lesion, after abortion, after normal vaginal delivery or after caesarean section. Some women had also been prescribed antibiotics by their doctor while being pregnant:“I had vaginal bleeding while being pregnant in the 4th month. I was admitted in the hospital, and I got the intravenous (IV) line. The doctor also gave me 1 flacon of antibiotics through IV. Usually, I took amoxicillin or ampicillin and paracetamol.” “The doctor gave antibiotic to patients when suturing the caesarean section, vaginal bleeding while being pregnant or abortion after discharging from the hospital.” “I had been taking antibiotics when I was 3–4 months pregnant due to heavy headaches and vertigo. I went to the army health clinic then I was diagnosed that the leukocytes in blood vessels had risen and the doctor prescribed ampicillin for 7 days. Though I stopped on the 3rd day because the symptoms were recovered well.” Participants expressed that they were somewhat reluctant to take antibiotics during pregnancy, concerned about the risk of congenital malformation. This might also explain non-adherence to antibiotic regimens, as below:There is an advantage of using antibiotic to kids, it is treating their sickness such as pneumonia. Last time I brought my kid to Mittaphab hospital, doctor prescribed antibiotic and my kid recovered within three days.” “The women felt however that children should be given antibiotics to fasten recovery.“My kid took antibiotics when he had pharyngitis or pneumonia. I bought antibiotic in the form of powder that costed about 10,000 LAK, but this medicine was not effective, however, the antibiotics from overseas that costed 40,000 LAK were more effective” Common reasons for giving antibiotics to their children, were fever, cough, pneumonia or pharyngitis. Amoxicillin, ampicillin, and erythromycin were the most used antibiotics. Some mothers also said they kept some in the house in case of illness.Note: 1US dollar = 10,000 LAK.“I understand what the doctors said, and I believe in the decisions of doctors because they are well educated in this field. I have heard some doctors said that the infectious diseases will last longer if we don’t follow doctor’s advice to take antibiotic correctly. Pneumonia will take longer time to be treated.” Most participants received information about antibiotics and antibiotic resistance from healthcare providers in public hospitals, private clinics and pharmacies and trusted the advice of the allopathic doctors:“I knew about antibiotic from my relatives as she used the antibiotic for her children” “I heard about using antibiotics from my friend during her pregnancy as the doctor prescribed antibiotic to her when she got some infectious diseases” Other sources of information related to antibiotic use were neighbours, friends, and family with experience in taking antibiotics. A few mothers said they had heard about antibiotics and antibiotic resistance from television and the internet.“I want to learn more about the causes of the reproductive tract infections such as leukorrea, sexually transmitted infections… and how to treat.” Most participants expressed an interest in receiving more information about the indications for antibiotic use, the benefits, and short- and long-term effects of antibiotic use for themselves and their children. Some mothers particularly wanted to learn more about the causes of reproductive tract infections, for example leukorrhea, sexually transmitted infections and how to prevent and treat the infections.“I want to learn about the side-effects of antibiotics in general and during pregnancy in every period; for examples, delivery and after giving birth. I want to learn more on the definition of infectious diseases, which ones are infectious diseases and which ones are not.” The women also wanted to learn more on antibiotic use during pregnancy, delivery, and postpartum period.“Source of information shall be provided by village speaker, Facebook, posters on roadside, hospitals, health centers, pharmacies, and village meetings.” Preferred sources of information were healthcare staff and village leaders. Other channels suggested by participants included television, radio, posters, websites and YouTube.This qualitative study explored perceptions and reported practices of pregnant women and mothers with children under two regarding antibiotic use and resistance. All participants had some understanding of antibiotic use and resistance, but misconceptions were evident and contributed to inappropriate antibiotic use. These modifiable practices are likely to be due to relatively low levels of education amongst participants and lack of consistent messages within the different information sources women used to obtain information. Most participants felt antibiotics are an effective treatment but had concerns about short and long-term side-effects of antibiotic use during pregnancy and early infancy. While self-medication was reported, during pregnancy and when their children were under 2 years, women stated they relied more on allopathic doctors, regarding treatment, including antibiotics use. The desire to learn more and the intrinsic desire for their children to thrive are encouraging and can be leveraged to promote appropriate antibiotic use  .As with previous studies , 27, 40,Pharmacies are often a first point of healthcare access in Lao PDR but are poorly regulated , 24, 32 In lower-middle income countries, younger children are often given medications by their parents without a doctor’s prescription . The preThe women in this study reported that during pregnancy and while seeking care for infants, they preferred to rely on the advice of allopathic doctors. As physicians and other healthcare workers play an important role in contributing to public knowledge of antibiotics, they are a powerful instrument for promoting appropriate antibiotic use and must prescribe appropriately and support patients to understand the importance of following the prescribed regimen. Health workers such as doctors and pharmacists can also educate people about the difference between bacterial and viral infections, and to better understand the concept of antibiotic resistance , 50, 51.The results of this study should be considered in the light of its limitations. As a qualitative study, caution should be taken in generalizing the findings beyond the sites where the data was collected. Nevertheless, the purpose of this exploratory study was not to make statistical inferences but to understand the cultural context of pregnant women and mothers’ perspectives of antibiotic use and resistance, providing a platform for further research. In addition, our study participants were mainly from Lao ethnic group and might not reflect the perspectives and experiences of women from different ethnic and cultural backgrounds within Lao PDR. Moreover, as the findings are based on the subjective experiences and self-reports of participants, the potential for recall and social desirability bias could not be discounted. Nevertheless, the study has provided useful information on perceptions and reported practices regarding antibiotic use and resistance among pregnant women and mothers of children under two. The findings gained from this study can be used to develop educational interventions  to promote appropriate antibiotic use.This study revealed that participants had misconceptions related to antibiotic use and resistance which might contribute to inappropriate antibiotic use. Participants, however, were concerned about antibiotic use during pregnancy and for their infants, and mostly sought advice from healthcare professionals in these contexts. Clear and consistent messages that can be disseminated using various channels, including trusted healthcare professionals, are likely to be the most effective. Messages can also potentially leverage the fundamental concern women have for their children. Awareness raising must also be complemented by efforts to address other determinants of inappropriate antibiotic use, including educating healthcare workers and pharmacists.


Fire and extreme heat environmental changes can have an impact on concrete performance, and as climate change increases, new concrete structures are being developed. Nano-silica and nano-calcium carbonate have shown excellent performances in modifying concrete due to their large specific surface areas. This review describes the changes in concrete modified with nano-silica (NS) and nano-calcium carbonate (NC), which accelerate the hydration reaction with the cementitious materials to produce more C-S-H, resulting in a denser microstructure and improved mechanical properties and durability of the concrete. The mechanical property decay and visualization of deformation of mixed NS and NC concrete were tested by exposure to high temperatures to investigate the practical application of mixed composite nanomaterials (NC+NS) to concrete. The nano-modified concrete had better overall properties and was heated at 200 °C, 400 °C, 600 °C and 800 °C to relatively improve the mechanical properties of the nano concrete structures. The review concluded that high temperatures of 800 °C to 1000 °C severely damaged the structure of the concrete, reducing the mechanical properties by around 60%, and the dense nano concrete structures were more susceptible to cracking and damage. The high temperature resistance of NS and NC-modified nano concrete was relatively higher than that of normal concrete, with NC concrete being more resistant to damage at high temperatures than the NS samples. Cement concrete has long been a structural material in civil engineering and continues to give rise to concepts such as Ultra-High-Performance Concrete (UHPC) ,2, RecycNS and NC are common nanomaterials in the market and are used in a wide variety of applications . NS has The addition of NS improves the performance of concrete and can significantly improve the defects that exist in the concrete itself . NS acce2 and nano-C-S-H shortened the induction period and the arrival time of the peak heat release rate of hydration, and nano-C-S-H also increased the peak heat release rate. The addition of nanoparticles and high temperatures led to the early production of hydrate layers [2 can improve the role of the thermal insulation capacity of concrete, due to the dense internal structure, increasing the specific heat capacity and reducing the thermal diffusivity [The effect of nanoparticles on the hydration of cement depends not only on the type of material but also on the temperature . A certae layers . Emergine layers . The scifusivity .Concrete in high-temperature areas is prone to structural damage and these damages seriously affect the service life of buildings. There is a need to conduct research and develop new nanomaterial-modified concrete with good mechanical properties and durability at high temperatures. An overview of many articles found that nanomaterial concrete technology continues to be researched and developed; NC and NS-modified concrete have a good evaluation in terms of economy and performance, because the constant deterioration of extreme high-temperature environments on concrete is a long-term impact, while whether nano concrete has a lot of resistance is based on the effects of fire and internal ambient temperature. By reviewing the latest prospective research on nano concrete, it is hoped that the argumentation will provide directional guidance for future research, give a characterisation of nano concrete, make concrete structures more sustainable and promote the development of concrete technology and industrial revolution. In the future, more applications of nano concrete will be seen.An overview of the microstructural changes, mechanical properties and durability of concrete mixed with NS and NC materials, as well as the mechanical decay and deformation in high-temperature environments is provided. Microscopic analysis techniques such as Scanning Electron Microscopy (SEM) and Differential Thermal Analyses (DTA) are used to reveal the internal morphology of the concrete. The paper reviews the changes in the mechanical properties of nano concrete in terms of compressive strength, flexural strength and durability properties such as water permeability, depth of carbonation and self-shrinkage. The final section further demonstrates the residual macro-mechanical properties and visualisation of the decaying deformation in the appearance of nano concrete after a high temperature, visualising the colour change and crack expansion of nano concrete after heating by means of content inference and picture synthesis. This review aims to investigate the effects and patterns of composite nanoparticles on the overall properties of concrete and to promote experimental research on nanocomposites (NC+NS). The following 2−4 reacts with Ca2+ to produce C-S-H seeds, which undergo secondary hydration with Ca(OH)2 to form C-S-H gels. NC is a modifier that increases the order of CH arrangement, filling tiny voids to produce a denser cement matrix. A certain amount of nanomaterials are mixed into concrete instead of cement to accelerate the hydration and heat of the hydration reaction of cement, which helps to fill the pores in concrete. This reduces the increase in the volume and denseness of the tiny pores in concrete . NS can A microscopic SEM examination of the composite concrete specimens after curing indicated the internal microstructure and the degree of hydration reaction ,46. FromZ. hang Wang et al.  concludeGuler, Türkmenoğlu et al.  analysed3 depletion in the 3% NS and cement binary composite samples.The incorporation of NS particles at optimum dosing levels improved the structural and mechanical properties of concrete. The addition of NS increased the hydration of the cementitious material and reduced and depleted the CH content; the extent of the change can be seen in the X-ray Diffraction (XRD) patterns in The rheological properties of concrete have been evaluated accordingly by many scholars in the configuration of concrete. As NS and NC are in the nanoscale (0–100 nm) range, very small nanoparticles fill the pore structure of concrete. Nano concrete exhibits greater water absorption and a larger specific surface area, which reduce compatibility and flowability . NS and The dynamic mechanical properties of concrete were tested by the compressive strength test, flexural strength test and splitting strength test, etc. The test process of casting cubic, rectangular and cylindrical samples can visualize the trend of a strength change of different kinds of nanomaterials in different shapes of concrete after different temperatures . The incAs shown in 2CO3 and NC, and that NC was effective in mitigating the loss of the 28-day combined strength of UHPC. After modification, the UHPC combined strength and flexural strength increased by about 68% and 38%, respectively, over the control sample. The optimum dosages of 3–4% NC were obtained, and 2% and 3% of NS and NC, respectively, influenced the heat of hydration and compressive strength of the cementitious material. The addition of 2% NS had the most significant effect on the mechanical properties of the concrete [T. Wang et al.  concludeconcrete . The mecconcrete . The comAccording to many studies, there was no significant difference in the amount of NS and NC replacement cement, with an overall range of 0% to 3%. The findings in 2O3 can reduce the weight loss during drying and the capillary permeability coefficient during water absorption. By increasing the nanomaterial content, the amount of charge passing through the sample is increased and the resistivity is subsequently reduced. Nanomaterial contents greater than 1% tend to agglomerate during hydration with the cementitious material, making the concrete less durable. Smaller admixtures allow for more uniform dispersion of nanoparticles and 0.5% nano concrete has better durability. In addition, the addition of NC also reduces the shrinkage of the concrete to a large extent. NS and NC contribute significantly to the improvement of the durability properties of concrete. The addition of 1%, 2%, 3% and 4% of the cement admixture of both NS and NC can reduce the water absorption of concrete. NC makes the cement matrix microstructure denser and produces more hydration products to improve the compressive strength and durability of concrete ,92,93. NBased on the durability studies carried out by many scholars, the water absorption, hazardous material permeability and freeze-thaw resistance of nano concrete are demonstrated, and 2 nanoparticles resulted in a lower cumulative heat of reaction than the addition of NC until 40 h into the reaction, with the results showing a higher heat of reaction by the end of 80 h. The relative increase in the cumulative heat of reaction for NS addition can be seen in Experiencing a fire or extreme high-temperature weather, through a series of reflections, the internal properties of concrete may change dramatically, by setting different high-temperature environments and simulating the state of concrete structure analysis, to derive the excellent medium difference of different nanomaterial admixture concrete, so as to develop the analysis ,107. ManNanomaterials mixed with other media, such as fibres, will have different mechanical property degradations in high-temperature environments. L. Wu et al.  investigYonggui et al.  studied NS and NC can improve the flexural properties of concrete in different temperature environments. By comparing 25 °C and 600 °C high temperatures, mixing NS improved the flexural strength and energy absorption capacity of the material. The flexural strength of NS mixed with 1.5% increased by 27% over the control sample in a room temperature environment, and by 21% after 600 °C high temperature. Unlike the NS modification, the addition of 3.0% NC was better at high temperatures than at room temperature. The flexural strength of concrete at an ambient temperature of 25 °C increased by 9% over the control sample and could be increased by 23% at 600 °C . It is wThe ambient environment of NS and NC makes the interfacial transition zone of concrete denser and holds with the aggregates to form higher mechanical properties . In contBased on the effects of changing global climatic extremes and fire on concrete structures, NS and NC modified the concrete microscopically, with the microfilling of pores, volcanic ash and hydration resulting in a more dense internal structure. The mechanical properties of the modified concrete were analysed by a nano-analysis and the compressive properties, splitting properties and flexural properties were improved, respectively. The durability performance varied slightly with the tested parameters, and where environmental conditions were harsh, the NS and NC admixtures needed to be increased to achieve a good balance. Based on the existing basic research, the review article derives the residual properties and decay deformation of NC and NS-modified concrete after high-temperature environments. Based on the review, it was found that the mechanical properties of NS and NC-modified nano concrete after high-temperature effects were not significantly affected at 200 °C, the mechanical properties decreased at 400 °C and the strength loss of the concrete accelerated as it continued to increase. Some authors such as Khan et al.  evaluateBoth NC and NS cause a hydration reaction within the concrete and at the same time fill the voids in the concrete, making the internal structure denser, as can be seen from the SEM, TGA and Infrared Spectroscopy Analysis.The mechanical properties of the concrete incorporated with the composite nanomaterial NC+NS are significantly enhanced, with a corresponding increase in mechanical properties under different high-temperature environments. The results of the review study show that concrete samples heated at different high temperatures such as 200 °C, 400 °C, 600 °C and 800 °C have higher strengths than the control concrete. The mechanical strength of the concrete does not change significantly in the 200 °C range, and the strength decreases from 200 °C to 600 °C. After high-temperature heating at 800 °C, the mechanical strength of concrete loses about 60%.The dense NC and NS materials reduce the water permeability, depth of carbonation and self-shrinkage of the concrete, contributing to the development of durability.Mixing of NC and NS materials can improve the thermal insulation of concrete structures, with NS having a more pronounced effect than NC. This facilitates the development and application of special insulation features for concrete.After continuous high temperatures, from 200 °C to 600 °C, the surface colour of NC and NS concrete gradually changes from grey to white, and from 600 °C to 1000 °C the colour gradually darkens to brown. The surface of the concrete starts to show obvious cracks at 600 °C and the concrete is more severely damaged at 800 °C, with the phenomenon of peeling of the skin. Concrete mixed with NC and NS has more severe appearance damage than normal concrete due to being denser and more closed, with temperatures exceeding 600 °C. In addition, the structural damage to the concrete is more severe as the time at high temperatures increases.By reviewing this research for the properties of nanomaterial in concrete, the decay deformation pattern of NC and NS in a high-temperature environment was obtained and the following conclusions can be drawn:Mixed nanomaterials have been shown to be effective in improving the mechanical strength of concrete, and long-term mechanical property changes in concrete are worth tracking using NC and NS studies. The durability of nano concrete structures after high temperatures necessitates further research as concrete is exposed to high temperatures for long periods of time through global warming. New properties of nanostructures are being developed to withstand the sustained high temperatures of fires, shielding against radiation, and nanomaterials are expected to be used in critical structures. An overview drives the experimental research on composite nano (NC+NS) concrete and some new conclusions will be drawn.


Supraglottic stenosis is a rare symptom, particularly in fibroinflammatory multifocal diseases, such as IgG4-related disease (IgG4-RD). There is still an inconsistency in the diagnosis of less-common locations of IgG4-RD, which causes a delay in the diagnosis and treatment. Our paper aims to analyze different aspects of IgG4-RD presenting as supraglottic stenosis, including the possible overlap with ANCA-associated vasculitis. We compare the usefulness of the recently revised ACR/EULAR and Comprehensive criteria and discuss treatment options. The review was performed according to PRISMA guidelines using the MEDLINE Pubmed and Scopus databases. The analysis includes nine papers describing supraglottic laryngeal stenosis in 13 patients. Furthermore, we present a case of a woman with ongoing supraglottic stenosis presenting with cough, temporary dyspnea and stridor as the symptoms of localized IgG4-RD. At the time of writing, the patient remains in remission while receiving treatment with cyclophosphamide and methylprednisolone. The symptoms of supraglottic localization of IgG4-RD may be severe; however, at that point, clinicians should suspect autoimmune etiology and attempt to modulate the autoimmune response instead of performing dilatation surgery—the effects of which may not result in extended intervals between interventions. The ACR/EULAR criteria show great specificity; however, when IgG4-RD is presumed, the specific treatment should be implemented. IgG4-related disease (IgG4-RD) is a relatively new fibroinflammatory clinical entity with a history dating back to 2003. Previously, the disease was referred to as autoimmune pancreatitis, retroperitoneal fibrosis, Kuttner tumors, Mikulicz’s disease, inflammatory pseudotumor, aortitis, Riedel thyroiditis and lymphadenopathy . The disHelicobacter pylori have been discussed .,17.16,17We identified a total number of 100 articles. After careful consideration, nine of them were included in our analysis . Table 1The chosen studies describe eight male and five female patients. One study included a child without defined age. In the adults’ group, the youngest patient was 29 years old, and the oldest was 76 years old. Female patients were reported to be more inclined to head and neck manifestations of the disease, which was not confirmed in the analyzed group of supraglottic stenosis . The mos’s syndrome, vasculitis, sclerosis cholangitis, rheumatoid arthritis, hypothyroidism and Helicobacter pylori infection, which cover the abovementioned IgG4-RD triggering factors. All the subjects were examined with flexible nasolaryngoscopy, which typically confirmed supraglottic and interarytenoid fibrosis, ulcer or diffuse swelling of the region, cicatricial narrowing and, in some cases, limited vocal fold mobility. Laser excision was performed in five cases; however, it had to be followed by another surgery. It was the pharmacological treatment that finally gave resolution of symptoms. The non-surgical approach included a wide range of medications: glucocorticoids, immunomodulatory drugs and cytostatic drugs; however, each of them is recommended in systhemic IgG4-RD [Several co-morbidities were described, including Sjogren IgG4-RD ,14,15,20A 72-year-old female patient L.G. presented with temporary dyspnea at rest, cough and globus symptoms. She had a 15-year history of hypothyroidism and glaucoma. No other significant diseases in her past or in family were described.At presentation at the Department of Otolaryngology, Head and Neck Oncology at the Medical University of Lodz, L.G. had severe supraglottic stenosis revealed in flexible nasolaryngoscopy. The laryngeal inlet was narrowed, and we observed thickening of the aryepiglottic folds, hypertrophy of posterior commissure with a tissue overhanging to the glottis and limited mobility of the vocal folds .We excluded cancer, injuries, iatrogenic causes, bullous disease, pemphigus vulgaris and other common causes of stenosis. Biopsy taken in local anesthesia from the oral cavity mucosa did not reveal either IgG immunoglobulin deposits or C3 complements. However, a biopsy taken from the laryngeal lesion revealed nonspecific active chronic inflammation. A direct immunofluorescence test, antibodies on the substrate of stratum spinosum and transitional epithelium were unrevealing, opposite to positive cANCA antibodies (1:40) (PR3 ++) and positive SES ANA (1:640) with the parepidermoidal normotypic epithelium. This led to a provisional diagnosis of a local form of ANCA-associated vasculitis (granulomatosis with polyangiitis).The patient’s condition initially improved after the methotrexate (20 mg/week) and prednisone (5 mg/day—starting from glucocorticoids pulses 3 × 1000 mg methylprednisolone and then 30 mg prednisone in descending doses). Full mobility of the larynx was restored. Due to poor tolerance and severe gastric complaints after 2 months of treatment, the methotrexate was changed to 150 mg azathioprine daily.Despite the initial improvement, the dyspnea gradually worsened over the next several months. The patient was re-admitted to the hospital with stridor and dyspnea. Endoscopic examination revealed stenosis of the upper airway with edematous posterior commissure of the larynx, which immobilized the arytenoid cartilages and impaired the vocal fold mobility C,D. The The histopathological examination of the sections from the supraglottic area revealed massive, subepithelial, mononuclear inflammatory infiltration A. Also, A definite diagnosis of localized IgG4-related disease was made . MultifoDiagnosis of IgG4-RD may be ambiguous, particularly when a non-specific and not common organ is affected. Currently, specific diagnostic criteria for laryngeal localization of IgG4-RD do not exist, as clinical trials are impossible due to the rare incidence and the fact that contemporary knowledge is typically based on case reports. We analyzed each case included in the study in terms of the Comprehensive and new ACR/EULAR criteria. Of 13 patients reported in the literature and patient L.G., nine had a probable diagnosis of IgG4-RD, and only four were definite. Some of those “probable” diagnoses were caused by an absence of IgG4 serum level, which may be either at a normal level or not assessed. On the other hand, it may disrupt an accurate diagnosis, as IgG4 serum levels are not routinely performed in hospitals, and its concentration may be elevated in numerous diseases .Nine patients with “definite” or “probable” IgG4-RD fulfilled the ACR/EULAR criteria for IgG4-RD diagnosis and gained 20 or more points . A similIgG4-RD is equally under- and over-diagnosed, as reported by Cheuk. Under-diagnosis results from low awareness of the disease, while over-diagnosis may be an effect of the increased level of IgG4+ cells .The disease is likely to form tumefactive lesions, and, in the head and neck region, it may be misdiagnosed as ANCA-associated vasculitis or malignancy. Furthermore, the authors hypothesize that IgG4-RD and ANCA-associated vasculitis in rare cases may overlap, particularly owing to the fact that they share a clinical and histopathological features ,26. The The distinction between IgG-RD and ANCA-associated vasculitis is crucial because these diseases have different prognoses and slightly different treatment approaches. This indicates the necessity to follow ACR and Chapel Hill for ANCA-associated vasculitis as well as IgG4-RD criteria to properly diagnose the diseases ,28.GPA affects patients at any age, with predominance in the sixth and seventh decade of life and can be localized in the larynx and trachea region; however, it often coexists with glomerulonephritis. In diagnosis, the presence of ANCA antibodies and histological examination of the fibrosis are necessary. A detailed scheme was suggested by the American College of Rheumatology . Mild foThe literature on the subject reports one case of a pediatric patient with a laryngeal lesion and finally diagnosed with Churg–Strauss syndrome. The lesion involved both vocal folds and the interarytenoid area. The mass was surgically removed but recurred after several months . NonetheA study from 2012 examined the prevalence of IgG4+ cells in biopsies from GPA patients with manifestation in the head and neck region. They found that 8 out of 26 probes revealed an increased number of IgG4+ cells. Hence, granulomatosis with polyangiitis may be misdiagnosed as the inflammatory background, fibrosis and high proportion of IgG4+ plasma cells is present in both diseases. Another study described that the IgG4-positive cells may be present in 38% of GPA cases in the head and neck region . A case In the case of L.G., the diagnosis of GPA with increased IgG4+ cells could have been made unless the criteria described in Treatment varies in those two entities. In ANCA-associated vasculitis, the treatment is standardized leading to a low relapse rate. This treatment includes cyclophosphamide or rituximab with glucocorticoids . On the Supraglottic laryngeal stenosis stands for 3% of all laryngeal narrowing cases; however, due to the rare incidence, some authors do not confirm the prevalence numbers . The mosTreatment typically includes endoscopic surgery, open surgery or a combination of both. In severe cases, and if the final treatment is delayed, patients may need a tracheostomy ,44. As tIt is currently discussed in the literature whether the surgical approach used as the first line of treatment in stenosis of unknown origin is best for the patient. Some authors underline the necessity for immunological system modulation rather than mechanical treatment . In the However, an open procedure significantly increases the risk, particularly when patients suffer from other co-morbidities compared to endoscopic procedures. On the other hand, the goal of extending the interval between interventions was achieved . In the Other patients were finally treated with prednisolone, rituximab, azathioprine or methotrexate, which covers the recommended treatment in IgG4-RD.  The detaPatients presenting with supraglottic laryngeal stenosis should have biopsies performed whenever possible and have those examined in search of IgG4-RD features. An appropriate workup can lead to a higher ratio of decannulation and reduce the voice and swallowing compromise. Furthermore, this may lead to a decrease in patient exposure to unnecessary surgeries. Laryngeal swelling, dyspnea and hoarseness may be the first isolated symptoms of IgG4-RD.Clinicians should mind both the Comprehensive and 2019 ACR/EULAR criteria, as the first encourages a reduction in unnecessary surgical procedures in favor of the pharmacological approach and the latter reduces false-positive diagnoses, as IgG4 may easily resemble several autoimmune diseases. The goal for patients with supraglottic stenosis in ongoing IgG4-RD is to extend the period without intervention, and it appears that this may be achieved by immunomodulatory drugs.


With the wide application of carbon fiber reinforced polymer (CFRP) plate, used for strengthening existed concrete structures, the prestressing technology of CFRP plate is becoming a hot topic, in order to sufficiently develop its high-strength peculiarity. In this paper, a full-scale hollow-section beam with length of 16 m taken from an old bridge which was in service for about 20 years was first examined for existed cracks and repaired by filling epoxy adhesive, and then the beam was strengthened with prestressed CFRP plates. The CFRP plates were tensioned and fixed with flat-plate anchorages at ends and bonded with adhesive on the bottom surface of the beam. The strengthened beam was experimentally studied using a four-point test to measure the concrete strain along the height of the mid-span section and the mid-span deflection. The finite element model of the strengthened beam was verified by the comparison of test results and used for an extending study of parametric analysis considering the effect of the length and amount of CFRP plates. Results indicated that with an increase in the length and amount of CFRP plates, the mid-span deflection of the beam decreases with the increased cracking resistance and bearing capacity, while the ultimate failure mode transfers from the under-reinforcement to the over-reinforcement. Prestressed concrete hollow-plate bridges are commonly constructed in highway and railway engineering with the advantage, including relatively lightened self-weight, of fast construction using cast-in-situ concrete with assembling of precast members ,2,3. In Regarding the strengthening methods for concrete structures, strengthening with fiber reinforced polymer (FRP) composites including carbon FRP, glass FRP and hybrid FRP, is a well-accepted technique due to its lightweight, high-strength, and good corrosion resistance ,10,11,12In order to develop the advantage of high-strength with a large stiffness of carbon FRP (CFRP), the prestressing method has been created to increase the strength utilizing efficiency of CFRP with a rational economy for improving the strengthening effect on reinforced concrete beams ,29,30,31Moreover, experimental research on full-scale reinforced concrete beams strengthened with prestressed CFRP plates have been performed. Wang et al.  conducteTo enrich research on the loading behaviors of full-scale reinforced concrete members strengthened with pressed CFRP plates, a full-scale hollow-section prestressed concrete beam was removed from an old bridge in operation for about 25 years, and transported to the lab in Zhengzhou, China, about 150 km from in-situ. The status of the beam was firstly detected to determine the properties of the concrete and the bearing performance of the hollow beam, and then the cracks were repaired by filling the epoxy paste to ensure the entirety of the beam. After that, the beam was strengthened by using prestressed CFRP plates, and the loading performance of the strengthened beam was experimentally studied by a four-point test. With the experience of using finite element (FE) analysis for the simulation of the loading process on reinforced concrete structures ,38, the Due to the limits of the experimental study, an extending parametric study was conducted to get a wide applicability of the experimental results. The effects of the length and the amount of prestressed CFRP plates on the flexural properties of prestressed concrete beams were analyzed using the FE model. Results provided a scientific basis for the strengthening technology of prestressed CFRP plates when applied to improve the loading capacity of old bridges, and the results could also be beneficial for similar projects.s5 prestressed steel wire. A total of 4ф12 plane hot-rolled steel bars were placed in the tensile zone, 6ф16 + 3ф12 plane hot-rolled steel bars were placed in the compressive zone, and ф6 plane hot-rolled steel stirrups in spacing from 150 mm to 200 mm were used to form a reinforcement skeleton. The concrete cover of the outside reinforcements was 28 mm.The prestressed concrete hollow-section beam investigated in this study was removed from an old simple-support assembly precast prestressed concrete bridge. This bridge was designed in accordance with China code JTG D62-85 , completDue to the long-term service of this old bridge, several cracks appeared on the side and bottom surfaces of the proto beam, as shown in 4 MPa.According to the specification of China code JGJ/T 23, the rebound method was used to inspect the concrete compressive strength . Ten zon5 MPa.After the test of the strengthened beam, samples of prestressed steel wire were taken from the ends of the beam to measure the mechanical properties according to China code JGJ/T 152 . ResultsTo maintain the integrity of the proto beam, the existed cracks should be repaired before strengthening. Based on the specification of China code GB 50367 , the epoAlthough the existing cracks were repaired by filling epoxy adhesive, it was hard to avoid the stiffness degradation of the proto beam. To evaluate the degree of stiffness degradation, a four-point bending test with a pure bending length of 3.6 m was conducted, as presented in Two concentrated loads were applied step by step on the top surface of the repaired beam. As shown in Based on the specification of China code JTG 3362 , the maxP0. The loading step followed as 0.2, 0.4, 0.6, 0.8, 0.85, 0.9, 0.95, and 1.0 times the target load P0.In this study, the target load was determined with the bending moment at the serviceability limit state. Therefore, each concentrated load on the top surface of the repaired beam was 109.8 kN, denoted by Bss of the repaired beam was obtained by the formula based on the test mid-span deflection af,α is coefficient, the value of α is 0.038 in this test; l is the span of the repaired beam, taken as 15.6 m; and af is the mid-span deflection at serviceability limit state.The stiffness degradation of the repaired beam was quantified by the following method. The stiffness Bs0 of the proto beam without degradation was calculated according to the formula specified in China code GB 50010 [Es is the modulus of elasticity of the longitudinal tensile steel bars; αE is a ratio of the modulus of elasticity for steel bar to concrete; ρ is the reinforcement ratio of longitudinal tensile steel bars; ftk is the tensile strength of concrete; ρte is the reinforcement ratio of longitudinal tensile steel bars in an effective sectional area; and σsq is the tensile stress of longitudinal tensile steel bars across cracks under the service load.The stiffness GB 50010 ,(2)Bs0=βf was obtained as follow,Therefore, the stiffness degradation coefficient βf = 0.73. This degradation was applied to the FE model of the repaired and strengthened beams by reducing the tensile strength of concrete in the pure bending segment. That is, the tensile strength of concrete was taken as 0.73 times 1.60 MPa as 1.17 MPa.Based on the test, the mid-span deflection was 54.1 mm at the serviceability limit state. Substituting the values of other parameters into above formulas, it was calculated that 5 MPa, respectively. The tensioning stress was 720 MPa, 30% of the ultimate tensile strength of the CFRP plate. The effective prestress was 533.3 MPa which determined by deducting the prestress losses due to the anchor deformation and locking shrinkage, and the relaxation of CFRP plate. Therefore, according to the formula specified in China code GB 50367 [The prestressing of CFRP plates was used to strengthen the repaired beam. The design method specified in China code GB 50367 was applied to determine the amount of CFRP plates . Based oGB 50367 , the benAs shown in P0. After that, the load was exerted until the strengthened beam failed. The value of the targeted ultimate bending moment was that of the repaired beam plus the strengthened by prestressed CFRP plates, that is, 716.5 + 167.0= 883.5 kN·m.The strengthened beam was tested using the same loading method as presented in Nonlinear FE analysis of the repaired beam and the strengthened beam were made by using the ABQUAS software. The C3D8R element was used for concrete, which was a eight-node linear block with reduced integration. The S4R element was used for the CFRP plates, which was a four-node doubly curved thin shell with reduced integration. The T3D2 element was used for the steel bars, and the nonlinear spring element Spring2 was used to simulate the bond-slip between prestressed steel wire and concrete. The cohesive contact was used to simulate the interfaces between CFRP plates and concrete. In view of the sectional dimension of the beam, most of the concrete elements were divided in sizes of 100 mm. Those with the size of 50 mm were also used to simulate the hollow to reduce stress concentration.The concrete damage plasticity (CDP) model based on Lubliner et al. , Lee andConcrete tension:dt and dc are the tension and compression damage factors of concrete, respectively; Ec is the modulus of elasticity of concrete; ft,r and fc,r are the peak tensile and compressive stress, respectively; and εt,r and εc,r are the tensile and compressive strain corresponded to the peak stresses, respectively.Concrete compression:E1, E2, the rigidity module G12, G13, G23, and the Poisson coefficient Nu12. Test values of these parameters are presented in Prestressed steel wire was assumed to be a linear elastic material. CFRP plate was assumed to be an anisotropic material which can be specified in the model by “Lamina” material type ,50. The τf is the bond stress; s is the slip; and The interface between prestressed steel wire and concrete was nodal, connected using a tied contact element Spring2 . The bonThe traction-separation-based modeling was used to express the adhesive property of interface between CFRP and concrete. The bond-slip relation of a bilinear model is presented in δ is the effective opening displacement; τ is the effective traction; K0 is the initial stiffness; τmax is the shear bond strength; Gf is the fracture energy; and τmax and Gf are calculated by Formulas (12) and (13), respectively [Gta are the shear modulus and thickness of the epoxy; and fc is the prism compressive strength of concrete.Where ectively .(12)τmaThe boundary conditions were defined assigning displacements to the reference point of rigid bodies at the left and right of the model. As the left acted as pin support and the right acted as roller support, constraints on translation freedom in X, Y and Z directions and rotation freedom in X and Y directions were added at the left, while constraints on translation freedom in Y and Z directions and the rotation freedom in X and Y directions were added at the right.P0. It can be seen that the longitudinal strains of concrete linearly varied at the mid-span section of the repaired beam under loads below the cracking load of 0.6P0. Agreement presents well between experimental and FE analytical values at this stage. However, the tensile strains of concrete had a larger difference between the values of experiment and the FE analysis when the load was more than the cracking load. This resulted from the cracks due to the tensile stress exceeding the tensile strength of concrete, while the homogeneous tensile strength of concrete was disturbed by the repaired existing cracks in the repaired beam. Generally, the strain maintains a plane at the section of the repaired beam under service load P0.The experimental and predicted longitudinal concrete strains at mid-span section are shown in P0 at serviceability limit state are shown in P0) for the repaired beam.The concrete stresses distribution along span of the repaired beam under the cracking load and the load P0. As a result, many flexural cracks appeared in the pure bending sections, which was consistent with the experimental results.With the increase in load on the repaired beam, the tensile stress increased in certain tensile zone while the compressive stress increased in the compression zone of the pure bending sections. Due to the unloading effect of the cracked concrete which transferred the tensile stress from the cracked section to the uncracked section, the tensile stress at the BROWN area of the nephogram decreased to be a lower value, while the tensile stress at the web of shear-span near the loading section reached a higher value, which would lead to shear cracks see b. HoweveP0 were 54.1 mm and 53.9 mm, respectively. This indicates that a differential of 0.35% for the FE analysis, which is feasible to simulate the influence of the initial damage on stiffness by degrading the tensile strength of concrete.P0. It can be seen that the longitudinal strains of concrete linearly distribute at the mid-span section, and the results of the experiment agreed well with those of the FE analysis when the load was lower than the cracking load (0.73P0) for the strengthened beam. From the FE analysis, the strain at the mid-span section basically maintained a plane when the load was over P0 up until to the ultimate load. Therefore, the assumption of plane strain is adaptable to the bending section of the strengthened beam.The experimental and FE analytical strains of concrete at the mid-span section are shown in Comparing the concrete strain in P0 at serviceability limit state, and the ultimate load. The results are shown in P0). This was consistent with the experimental observation. The cracking load increased by 21.2% for the strengthened beam compared to the repaired beam. This verified that the prestressed CFRP plates helped arrest the cracking of the concrete.The stress distribution of concrete along the span of the strengthened beam was analyzed under the cracking load, the load P0, except for a lower tensile stress at the shear span of the strengthened beam due to the precompression at the tensile zone by the prestressed CFRP plates. Under the ultimate load, many more vertical cracks at the pure bending segment, accompanied with diagonal cracks in the shear span, appeared on the strengthened beams. The maximum compressive stress at the compression zone reached 24.2 MPa, which was close to the compressive strength 24.7 MPa of concrete. This corresponded to the damage to concrete at the compression zone, as shown in Similar nephograms existed on the strengthened beams and the repaired beam under the load P0, the stress linearly varied with the increase in load. Then, the stress tended to increase quickly, due to the loss of the ability of the cracked concrete to bear tensile stress which was transferred to the CFRP plates and steel wires. The experimental and FE analytical stress of the CFRP plates were 953.7 MPa and 919.5 MPa at the ultimate load. This also demonstrated the validity of the FE model with a difference of 3.6% from the experiment.The comparison of the experiment with the FE analytical results of the tensile stress of prestressed CFRP plates, and the FE analytical results of the tensile stress of prestressed steel wire are presented in Due to the proto beam being taken from an old bridge, the real stress of prestressed steel wire was not available. The stress of the prestressed steel wire of the strengthened beam under its ultimate load was close to the tensile strength of 1600 MPa, using FE analysis. This means that the prestressed steel wire could reach the yield. Meanwhile, the stress of prestressed CFRP plates was lower than the tensile strength of 2400 MPa. This means no brittle fracture of CFRP plates could happen, which was consistent with the experiment where only CFRP plates in some segments unbounded from the concrete surface.As presented in P0 was 42.8 mm, which decreased by 20.6% in comparison with that of the repaired beam. This demonstrates that the prestressed CFRP plates effectively improve the stiffness of the strengthened beam, thereby improve the bearing capacity.The mid-span deflection of the strengthened beam under the load In this section, a parametric study is conducted in view of the impacts of the length and amount of prestressed CFRP plates on the loading performance of the strengthened beam in response of the cracking load, ultimate load, and the mid-span deflection.Based on the verification of test results with the FE analysis, the concrete in the compression zone of the strengthened beam was crushed after experiencing large deflections. In this process, of the many cracks appeared in the pure bending segment play a dominate role in the failure of the beam. Therefore, the bonding length of prestressed CFRP plates should be greater than the pure bending segment, i.e., 3.6 m. In this section, the bonding length of prestressed CFRP plates is set to 3.6 m, 7 m, 10 m, and 13 m, with the repaired beam as a reference. Four FE models are built on basis of It can be seen that the cracking and ultimate bending moment increase with the increasing bonding length of the prestressed CFRP plate. On the contrary, a decrease presents for the ultimate mid-span deflection. Compared with the repaired beam, the cracking moment and ultimate moment of the strengthened beam increase by 28% and 10.6%, respectively, while the ultimate mid-span deflection reduces by 12.3%. This indicates that a more beneficial effect takes place on the cracking resistance with the increase in bonding length of prestressed CFRP plates. Combined with the concrete stress nephograms, the area of tensile stress over tensile strength of concrete decreases with the increase in the bonding length of prestressed CFRP plates, although the length of tensile area along the span remains unchanged at about 10 m. This reduces the possibility of concrete cracking and confines the extension of cracks in the tensile area. Therefore, the depth of compression zone increases to improve the loading capacity, and the flexural stiffness increases to reduce the mid-span deflection.In practice, the bonding length of prestressed CFRP plates should be rational to cover the middle segment to confine the possible cracks of concrete. Due to being subjected to moving vehicle loads on bridges, it is better for the simply supported hollow-section beam to be completely strengthened in span by prestressed CFRP plates, except for a certain length near the ends to install anchors and for the convenience of tensioning operation.Six FE models were built to investigate the effect of the amount of prestressed CFRP plates on the mechanical properties of the strengthened beam with the repaired beam as a reference. The number of prestressed CFRP plates was set to 3, 6, 9, 12, 15, and 18, respectively. Each plate was the same as the experiment, with a width of 54 mm and a thickness of 1.4 mm. The bonding length was set to 13 m.P0 with a smaller deflection due to the enhancement of concrete cracking resistance. With the number of prestressed CFRP plates increased from 3 to 12, the cracking moment increases by 65.6% in total, and each prestressed CFRP plate increases the cracking moment by 7.3%. When the number of prestressed CFRP plates is no less than 12, the crack of concrete can be avoided for the strengthened beam at serviceability limit state.At the same time, the ultimate bearing capacity can be obviously improved with the failure characterized by the under-reinforced mode transferred to the over-reinforcement mode. When the number of prestressed CFRP plates increases from 3 to 12, the ultimate moment linearly increases by 23.8% in total, while the mid-span deflection decreases by 31.5%. When the number of prestressed CFRP plates is no less than 12, the stress of prestressed steel wire is less than the designed tensile strength of 1280 MPa at the ultimate bearing capacity. This means that the prestressed steel wire cannot yield due to over-reinforcement by prestressed CFRP plates. In this condition, the increase in the ultimate bearing capacity of the strengthened beam is achieved at the expense of deformation ductility, and the ultimate moment.In practice, to maintain sufficient ductility of the strengthened beam with obviously deflection before failing ,43, the (1)Although several concrete cracks appeared on the proto beam, the concrete and the prestressed steel wire worked well without obvious degradation and steel erosion. After the concrete cracks were repaired with epoxy adhesive, the degradation coefficient of flexural stiffness was determined as 0.73, based on the experimental study of the repaired beam by four-point bending test.(2)FE models are built for the analyses of the repaired beam and the strengthened beam, including the concrete strain at mid-span section, the stress of the concrete and the prestressed steel wire, and the stress of prestressed CFRP plates, as well as mid-span deflection. Good agreement between the test and FE analytical results of the loading response demonstrates the validity of the FE model for simulating the strengthened beam with prestressed CFRP plates.(3)A parametric analysis indicates that the increase in length and amount of prestressed CFRP plates improves the serviceability and the ultimate bearing capacity of the strengthened beam. Additional benefits to improve the flexural stiffness come from the confinements of prestressed CFRP plates to the existed concrete cracks and by increasing the length of prestressed CFRP plates to the newly appeared concrete cracks, and the improvement of concrete cracking resistance by increasing the amount of prestressed CFRP plates.(4)It should be noticed that with the increased amount of prestressed CFRP plates, the failure mode of the strengthened beam can be transferred from the under-reinforcement to the over-reinforcement. This leads to an expense of deformation ductility of the strengthened beam.In this paper, a hollow-section beam was taken from an old simple support assembly precast concrete bridge which is normally in service for about 25 years. Experimental studies were carried out for the status detection of proto beam, the repair of existed concrete cracks, the strengthening by prestressed CFRP plates, and the loading tests for the repaired and strengthened beams. The FE model is verified by the test results, and used for extending parametric analysis. Conclusions can be drawn as follows:


Increasing evidence has demonstrated that circular RNAs (circRNAs) are implicated in cancer progression. However, the aberrant expression and biological functions of circRNAs in clear cell renal cell carcinoma (cRCC) remain largely elusive.Differentially expressed circRNAs in cRCC were filtered via bioinformatics analysis. Aberrant circPOLR2A expression was validated in cRCC tissues and cell lines via qRT-PCR. Sanger sequencing was used to identify the backsplicing site of circPOLR2A. In vitro and in vivo functional experiments were performed to evaluate the role of circPOLR2A in cRCC malignancy. RNA pull-down, mass spectrometry, RIP, FISH and immunofluorescence assays were used to identify and validate the circPOLR2A-interacting proteins. Ubiquitination modification and interaction between proteins were detected via Co-IP and western blotting. The m6A modification in circPOLR2A was validated by the meRIP assay.Bioinformatics analysis revealed that circPOLR2A was highly expressed in cRCC tissues and metastatic cRCC tissues. CircPOLR2A expression was associated with tumor size and TNM stage in cRCC patients. In vitro and in vivo functional assays revealed that circPOLR2A accelerated cRCC cell proliferation, migration, invasion and angiogenesis, while inhibiting apoptosis. Further mechanistic research suggested that circPOLR2A could interact with UBE3C and PEBP1 proteins, and that UBE3C could act as a specific ubiquitin E3 ligase for the PEBP1 protein. The UBE3C/circPOLR2A/PEBP1 protein-RNA ternary complex enhanced the UBE3C-mediated ubiquitination and degradation of the PEBP1 protein which could inactivate the ERK signaling pathway. Rescue experiments revealed that the PEBP1 protein was the functional downstream target of circPOLR2A. Furthermore, m6A modification in circPOLR2A was confirmed, and the m6A reader YTHDF2 could regulate circPOLR2A expression.Our study demonstrated that circPOLR2A modulated the UBE3C-mediated ubiquitination and degradation of the PEBP1 protein, and further activated the ERK pathway during cRCC progression and metastasis. The m6A reader, YTHDF2, regulated circPOLR2A expression in cRCC. Hence, circPOLR2A could be a potential target for the diagnosis and treatment of cRCC.The online version contains supplementary material available at 10.1186/s12943-022-01607-8. Renal cell carcinoma (RCC), arising from genetic changes, accounts for approximately 2% of all adult malignancies . Among dCircular RNAs (circRNAs) are covalently closed RNA molecules originating from precursor mRNA via noncanonical backsplicing in eukaryotes , 10. ThePhosphatidylethanolamine Binding Protein 1 (PEBP1) is a member of the phosphatidylethanolamine-binding protein family . InitialN6-methyladenosine (m6A), an epigenetic modification of RNA, is involved in RNA metabolism, including transcription splicing, translation and degradation . It was In the present study, we screened out a circRNA originating from POLR2A, hsa_circRNA_092437 (circBase ID: hsa_circ_0000741), named circPOLR2A, by analyzing the expression profiles of circRNAs in cRCC. Higher expression of circPOLR2A was verified in cRCC compared to matched adjacent normal tissues. We demonstrated that circPOLR2A promoted cRCC progression by mediating the ubiquitination of PEBP1 and activating the Raf1/MEK/ERK pathway. In addition, we further revealed that low expression of YTHDF2 contributed to upregulation of circPOLR2A in cRCC, which indicated that the expression of circPOLR2A was associated with m6A methylation modification. Our work suggested that circPOLR2A could act as an oncogene in cRCC progression and a potential target in the treatment of cRCC.All methods and materials are described in the p value < 0.01 and log2(fold change) > 3  repository: GSE100186, determining the expression profiles of circRNAs in 4 pairs of cRCC and nontumorous tissues ; GSE1378Next, we determined the 9 circRNAs in 32 pairs of cRCC samples and adjacent normal samples via quantitative reverse transcription PCR (qRT-PCR) with divergent primers. As shown in Fig. circPOLR2A is generated from exons 9–10 of the POLR2A gene on chromosome 17p13.1  Western blot indicated that no AGO2 protein was detected in the precipitates of RNA pull-down. (b) No significant change in PEBP1 mRNA expression was determined after circPOLR2A knockdown or overexpression.  Bioinformatics analysis of the CPTAC database suggested that PEBP1 protein levels were associated with cRCC tumor size (c), pathologic T stage (d) and Fuhrman grade (e) in the cRCC cohort. (f) Kaplan-Meier method and log-rank test confirmed that the PEBP1 protein level was a favorable prognostic factor in the cRCC cohort from CPTAC database.Additional file 3: Supplemental Fig. 3. Statistical analysis on the tube formation assay. (a) Total number of junctions in the tube formation assay for circPOLR2A knockdown or overexpression. (b) Total vessels length in the tube formation assay for circPOLR2A knockdown or overexpression.  Total vessels length in the tube formation assay for rescue experiments.Additional file 4: Supplemental Fig. 4.  Bioinformatics analysis suggested that YTHDF2 expression was associated with cRCC grade (a) and stage (b) in the cRCC cohort from TCGA database. (c) Western blotting detected the YTHDF2 level after circPOLR2A knockdown or overexpression. (d) The 4 putative m6A motifs of circPOLR2A predicted by the SRAMP prediction server. (e-h) The expression of METTL3 and YTHDF2 in cRCC cells transfected with siNC, siMETTL3 or siYTHDF2. 10e, 10f, METTL3 expression; 10 g, 10 h, YTHDF2 expression.  The schematic diagram illustrated the structure of circPOLR2A-WT (i) or circPOLR2A-MUT (j).Additional file 5: Supplemental Table 1.Additional file 6: Supplemental Table 2.Additional file 7: Supplemental Table 3.Additional file 8: Supplemental Table 4.Additional file 9: Supplemental Table 5.Additional file 10: Supplemental Table 6.Additional file 11. Supplemental Methods and Materials.