This paper proposes a physics-informed neural network (PINN) for predicting the early-age time-dependent behaviors of prestressed concrete beams. The PINN utilizes deep neural networks to learn the time-dependent coupling among the effective prestress force and the several factors that affect the time-dependent behavior of the beam, such as concrete creep and shrinkage, tendon relaxation, and changes in concrete elastic modulus. Unlike traditional numerical algorithms such as the finite difference method, the PINN directly solves the integro-differential equation without the need for discretization, offering an efficient and accurate solution. Considering the trade-off between solution accuracy and the computing cost, optimal hyperparameter combinations are determined for the PINN. The proposed PINN is verified through the comparison to the numerical results from the finite difference method for two representative cross sections of PSC beams. Most machine learning-based approaches for predicting the mechanical behaviors of structural systems require a huge amount of high-quality data for supervised learning. In many real-world engineering problems, such vast amounts of data are often unattainable on site, and limitations in data accessibility pose challenges for machine learning based on field data. To overcome these limitations, the physics-informed neural network (PINN) has gained attention for its capability of incorporating fundamental physical  knowledge, presented in the form of differential or integral equations, into machine learning ,5,6,7,8.The PINN addresses data limitation challenges by excelling in solving problems with uncertain or missing information  and holdThe PINN utilizes a feed-forward architecture combined with automated differentiation in artificial neural networks to train models that satisfy the governing equations, enabling the construction of physical prediction models with far fewer field data than traditional data-driven neural network techniques . The PINPINNs demonstrate constraints when handling issues that evolve sharp gradients or involve interconnected PDEs . As a coThe incorporation of adaptive activation functions into traditional PINNs has emerged as a compelling alternative ,44,45,46In the civil engineering industry, prestressed concrete (PSC) beams are increasingly used in construction due to their advantages over reinforced concrete (RC) beams in controlling cracking and minimizing long-term deflections . To desiThis paper presents a PINN-based approach for predicting the early-age time-dependent behaviors of a PSC beam considering interaction among concrete creep and shrinkage, tendon relaxation, and the changes of concrete elastic modulus. We introduce a novel closed-form integro-differential governing equation, formulated in terms of effective prestress force, to predict the early-age time-dependent behaviors of PSC beams. Contrary to the traditional finite element method, the PINN directly solves the integro-differential equation without requiring discretization. There exists a trade-off between solution accuracy and the computing time when solving the integro-differential governing equation through PINN. Balancing the importance of the solution accuracy and the computing cost, optimal hyperparameter combinations are determined for the PINN.The paper is organized as follows. (i) Multiple tendons of the PSC beam can be substituted with a single equivalent tendon as shown in (ii) The tendon is perfectly bonded with concrete; thus, section analysis for equilibrium between the tendon and the concrete is possible;(iii) The beam model follows the Euler–Bernoulli beam theory;(vi) The time-dependent behaviors of concrete include the change of elastic modulus, creep, and shrinkage;(v) The creep of concrete follows the standard linear model of viscoelasticity;(vi) The relaxation of the tendon is assumed to be intrinsic;(vii) The shrinkage of concrete occurs uniformly throughout the gross section of concrete;(viii) The net strain of concrete consists of elastic strain, creep strain, and shrinkage strain;(ix) The self-weight of the PSC beam is introduced at the moment of initial prestress transfer.In the simplified model for a PSC beam in The governing equations for time-dependent behaviors can be expressed as follows:(i) Axial-bending stress equation of concrete(ii) Constitutive equation of elastic strain of concrete (2)εely,(iii) Constitutive equation of creep strain of concrete (3)εcry,(iv) Net strain of concrete(v) Constitutive equation of elastic strain of the tendon(vi) Constitutive equation of relaxed strain of the tendon(vii) Net strain of the tendon(viii) Compatibility between the concrete and tendon at  otherwise mentioned hereafter. For example, substituting For the simplicity of formulation in (i) Dimensionless axial-bending stress equation of concreteIn a similar fashion, Equations (2)–(8) can be rewritten as follows:(ii) Dimensionless constitutive equation of elastic strain of concrete(iii) Dimensionless constitutive equation of creep strain of concrete(iv) Dimensionless net strain of concrete(v) Dimensionless constitutive equation of elastic strain of the tendon(vi) Dimensionless constitutive equation of relaxed strain of the tendon(vii) Dimensionless net strain of the tendon(viii) Dimensionless compatibility between the concrete and tendon at Because there are eight unknown dimensionless variables Integrating Equation (16) with respect to Substituting Equations (10) and (13) at Substituting Equation (13) into Equation (15) producesSubstituting Equation (17) with Equation (18) into Equation (20) results inSubstituting Equation (12) into Equation (21) producesIntroducing a prestress transfer coefficient Substituting Equation (24) into Equation (22) results in the following equation:Substituting Equation (13) into Equation (14), Substituting Equations (26) and (27) into Equation (25) produces an integro-differential equation for The numerical solution of Equation (28) for the dimensionless effective prestress force Letting constant time increment The derivative Substituting Equation (31) into Equation (30) producesSubstituting Equation (29) incorporated with Equation (32) into Equation (28) yields the finite difference equation of Equation (28)The numerical procedures to solve Equation (33) for The advantage of the PINN over conventional numerical methods lies in solving partial differential equations in strong forms without discretization through automatic differentiation. When the governing equation is expressed as an integro-differential equation like Equation (28), the PINN can be employed to solve it through automatic differentiation for integer-order derivatives and approximate integral operators based on Gaussian quadrature .Substituting The right-hand side of Equation (34) can be rearranged by separating the integral term from the non-integral terms as follows:The basic architecture of the PINN is illustrated in The PINN architecture shown in ode 1990 . The relode 1990  for low-The forward finite difference method elaborated in  DeepXDE  was adopFor each hyperparameter combination outlined in Three implications can be drawn from Similar to The optimal Based on the observation of The analysis of time-dependent behaviors of the PSC beam is conducted by using the forward finite difference method in the same way described in The dimensionless integro-differential governing equation is derived for the PINN. The governing equation accounts for time-dependent coupling among the effective prestress force and several factors including concrete creep and shrinkage, tendon relaxation, and changes in concrete elastic modulus.The fidelity of the PINN is determined by three primary hyperparameters: the number of nodes, the number of hidden layers, and the number of domains. Among these primary hyperparameters, the number of domains is the most critical factor for the accuracy and consistency of predicted solutions from the PINN. Optimal hyperparameter combinations for the PINN were determined by considering the trade-off between solution accuracy and computation time.The numerical results from the PINN with an optimal hyperparameter combination yielded satisfactory results, with absolute differences at 365 days in loss of prestress force being 0.027% and in all stresses at the top and bottom of the beam and the tendon location being less than 0.2% for PSC beams with a rectangular and an I-shaped section.The proposed PINN approach can effectively predict time-dependent behaviors of PSC beams, offering a promising alternative to conventional numerical methods. The proposed PINN can address the limitations of conventional machine learning by training deep neural networks that satisfy the governing equation associated with the complicated time-dependent behaviors of the PSC beam.In this study, a PINN approach was proposed to predict the early-age time-dependent behaviors of the PSC beam. The original contributions of this study are presented below:Future work will pursue a PINN to solve inverse problems such as estimating time-dependent material properties related to the concrete creep and shrinkage, and the relaxation of the tendon by using a limited amount of actual measurement data from the PSC beam. It is necessary to implement a more advanced PINN to solve the proposed integro-differential equation to improve the performance of the conventional PINN used in the present study. Future work will explore the incorporation of optimal adaptive activation functions ,44,45,46


Denosumab (DMAb) is a human monoclonal antibody used as an antiresorptive drug in the treatment of osteoporosis. Approval at a dosage of 60 mg every 6 months was based on the results of the randomized, placebo-controlled trial (FREEDOM). The design of this 3-year study included an extension for up to 10 years. Those who were randomized to DMAb continued on drug, while those who were randomized to placebo transitioned to DMAb. The 10-year experience with DMAb provides data on efficacy of drug in terms of reduced fractures and continued increases in bone mineral density (BMD). The 10-year experience with denosumab also provides information about rare complications associated with the use of DMAb, such as osteonecrosis of the jaw (ONJ), and atypical femoral fractures (AFF). This experience provided new insights into the reversibility of effects upon discontinuation without follow-on therapy with another agent. This review focuses upon prolonged treatment with DMAb, with regard to beneficial effects on fracture reduction and safety. Additionally, its use in patients with impaired renal function, compare its results with those of bisphosphonates (BPs), the occurrence/frequency of complications, in addition to the use of different tools, from imaging techniques to histological findings, to evaluate its effects on bone tissue. Denosumab (DMAb) is a fully human monoclonal antibody to the receptor activator of nuclear factor-κB (RANKL), inhibiting the development and activity of osteoclasts, followed by supression of bone resorption .DMAb is considered a relevant option in the treatment of osteoporosis owing to the corresponding beneficial outcome of DMAb administration – i.e., reduced risk of fractures . Rare side effects and risks related to the drug, such as osteonecrosis of the jaw (ONJ), atypical femoral fractures (AFF), and rebound effect should always be considered during clinical management ,2.In this narrative review, we address the evidence for the efficacy of DMAb in the treatment of osteoporosis.The Fracture Reduction Evaluation of Denosumab in Osteoporosis Every 6 Months (FREEDOM) study, published in 2009, led to the approval of DMAb as a treatment for osteoporosis. This randomized placebo-controlled clinical trial involved 7,808 postmenopausal women with osteoporosis  in which 60 mg of DMAb was administered subcutaneously every six months over a 36-month period. Exclusion criteria were women with other skeletal disorders, current or recent therapies for osteoporosis , T-Score of ≤ 4.0 in lumbar spine or total femur, with a history of at least one severe or two moderate vertebral fractures, or with serum levels of 25-hydroxyvitamin D < 12 ng/mL. The primary outcome index was the incidence of vertebral fracture after 3 years. Secondary outcomes were non-vertebral and hip fractures. The 36-month incidence of vertebral fractures identified by radiographs in the DMAb group was 2.3% vs. 7.2% in the placebo group, representing a 68% relative risk reduction (p < 0.001) with DMAb use. Similar results were also observed when evaluating the occurrence of multiple vertebral fractures and clinically evident fractures. DMAb also reduced the relative risk of hip fracture by 40%  and non-vertebral fracture by 20% . The drug significantly increased lumbar spine and total femur bone mineral density (BMD) , as well as reductions in bone turnover markers . There was no increase in the incidence of cancer, cardiovascular events, infections, or delay in the healing of fractures associated with DMAb. There was no case of ONJ during the 36-month follow-up  (estimated glomerular filtration rate of 15-29 mL/min). In all groups, DMAb was not associated with any significant changes in serum calcium or creatinine levels. Effectiveness in reducing fractures was pervasive from CKD stages 1 n = 842), 2 n = 4,069) and 3 n = 2,817). A slight reduction in CKD stage 4 (n = 73) was not significant  has also provided important microstructural data after DMAb exposure. It has the capacity to measure volumetric bone density, as well as cortical and cancellous bone microarchitecture ,21. Of tMore readily available but not as quantitatively reliable, as the bone biopsy and HR-pQCT, is the trabecular bone score (TBS), a software adaptation of lumbar spine Dual-energy X-ray absorptiometry (DXA) ,16,19,20These imaging modalities, taken together, and followed for up to 10 years, together with the reduction in fracture occurrence over this period of time, suggest that prolonged inhibition of osteoclastic activity by DMAb has overall a beneficial effect on bone quality ,17,19,21ONJ and AFF are rare complications of DMAb. Rapid reversibility of its actions is also a point of discussion.ONJ is a rare but serious complication of antiresorptive therapy, which can occur with either BPs or DMAb use. It is more commonly observed in patients with neoplasms treated with higher doses than used for osteoporosis ,24. In tEpidemiological data on ONJ are difficult to obtain, given its low prevalence, together with limitations in sample size and study design . The risAFF are typically subtrochanteric and occurring spontaneously or with minimal trauma. They typically involved diaphyseal insufficiency and may have delayed consolidation ,16,30. IAn analysis involving FREEDOM data and a hypothetical virtual placebo group demonstrated a satisfactory risk/benefit ratio with 10 years of DMAb therapy, observing 281 and 40 clinical fractures prevented for each episode of AFF and ONJ, respectively .Another possible complication associated with the use of DMAb is the so-called rebound effect. Its discontinuation, without follow-on use of another anti-osteoporotic drug, leads to an increase in bone turnover and a rapid decrease in BMD, leading to a potentially high risk of multiple vertebral fractures ,11,34. MIn conclusion, DMAb is an efficacious agent in the treatment of osteoporosis. Long-term therapy for up to 10 years has demonstrated continued efficacy with an acceptable safety profile for those at continued high risk. As with other therapies, its use requires monitoring with attention to inadvertent withdrawal and to the possibility of rare complications.


Phyllocoptes fructiphilus (Pf), both native to North America, has caused significant damage to roses over the last several decades. As cultural and chemical control of this disease is difficult and expensive, a field trial was established to systematically screen rose germplasm for potential sources of resistance. One hundred and eight rose accessions representing the diversity of rose germplasm were planted in Tennessee and Delaware, managed to encourage disease development, and evaluated for symptom development and viral presence for three years. All major commercial rose cultivars were susceptible to this viral disease to varying levels. The rose accessions with no or few symptoms were species accessions from the sections Cinnamomeae, Carolinae, Bracteatae, and Systylae or hybrids with these. Among these, some were asymptomatic; they displayed no symptoms but were infected by the virus. Their potential depends on their ability to serve as a source of viruses. The next step is to understand the mechanism of resistance and genetic control of the various sources of resistance identified.Rose rosette disease (RRD) caused by the rose rosette emaravirus (RRV) and transmitted by the eriophyid mite  Emaravirus rosae), was first reported in the western part of North America and has since moved to the eastern coast. Within the last couple of decades, it has emerged as the major disease in cultivated roses in North America [Rose rosette disease, caused by the rose rosette emaravirus  is a widespread threat in the USA to rose producers, landscapers, and consumers ,23,24,25and Asia .R. arkansana Porter, R. blanda Aiton, R. carolina L., R. californica Cham. and Schon., Rosa palustris Marsh, and R. setigera Michx.) and one Asian species (R. spinossisima (L.)) did not develop symptoms of the disease after grafting infected buds onto them and were considered resistant to RRD [R. arkansana Porter and R. woodsii Lindl., were reported as tolerant to the viral disease. Other species, including three from North America ) and 12 with Asian or European origins, appeared to be susceptible to RRD. These Eurasian species included R. banksiae Aiton, R. canina, R. dumetorum Thuill, R. eglanteria L., R. rubrifolia Vill., R. soulieana Crep., R. spinosissima var. altaica (L.) Rehd., R. montezuma Hum and Bonpl., R. multiflora Thunb., R. odorata (Andr.) Sweet, R. gallica L., R. hugonis Hemsl., R. villosa L., and R. wichurana Crepin. These species belong to eight different sections of the genus Rosa and thus represent wide genetic diversity. It should be noted that RRD resistance in one accession of a species cannot be extrapolated to all accessions of the species.The first reports about resistance to RRD among roses comes from grafting work, which indicated that several North American species  was reported to be resistant to the mite but not the pathogen. This included a wide range of diversity, including climbers, hybrid rugosas, floribundas, hybrid teas, miniature, polyanthas, one rootstock, and 14 Old Garden Roses . SubsequWork in Arkansas showed that the roses Bonica and Homerun (‘WEKcisbako’) were difficult to infect and that the rose Stormy Weather (‘ORAfantanov’) was not infected by either budding with an infected bud or by the feeding of infected mites . The flo1 populations with nine parents) as well as two connected tetraploid populations involving three cultivars (My Girl (‘BAIgirl’), Stormy Weather™, and Brite Eyes™ (‘RADbrite’)). A QTL located on linkage group (LG) 5 coincided in both diploid and tetraploid germplasm, had the greatest effect on RRD resistance (explaining 20 to 41% of the phenotypic variance), and was consistently detected in multiple datasets. The sources of the RRD resistance QTL on LG5 were traced back to the tetraploid variety Brite Eyes™ and the diploid TAMU breeding line M4-4, both with Rosa wichurana in their parentage. Additional minor QTLs were identified on LG1, LG3, LG6, and LG7.Recent research identified several genes/quantitative trait loci (QTL) for partial resistance to RRD ,29 in anPf [Pf was not able to reproduce on two rose species accessions (R. bracteata and R. carolina) or plants from related genera in the Rosaceae family.Mite reproduction studies on 32 modern rose cultivars have shown that modern roses are good hosts for Pf ,19,30, aRoses are one of the most popular plants in public gardens and commercial/residential landscapes. Ornamentals, including roses used in home, private, and public landscapes, promote human well-being, enhance air and water quality, reduce runoff and erosion, facilitate rain capture and stormwater management, reduce noise and dust pollution, and increase property values . The rosThe landscape rose market accounts for 35% of the roses sold, but this share recently decreased by 10–15% due to RRD. The estimated loss of rose sales from the RRD epidemic is USD 5–10 million annually. A recent introduction of RRD into a California production block resulted in expenses of over USD 1,000,000 to eliminate the threat and has led others to change their protocols at a high cost to prevent the disease from getting into their production blocks. Increased production costs drive up rose wholesale and retail prices while depressing rose sales in a market already being devastated by the fear of RRD. This is reflected in a 23% decrease in annual value (adjusted for inflation), a 33% decrease in the number of shrub roses produced, and 19% fewer businesses producing shrub roses from 2014 to 2019 .Since there is no chemical control of the virus, current control approaches include exclusion via the use of clean stock material, the decrease in inoculum/vector populations by roguing of symptomatic plants in and around the garden, the use of barriers to slow the vector’s spread, and chemical control of the vector . An apprThe objective of this field trial was to evaluate a suite of roses that represent the wide genetic diversity of the cultivated and wild roses for their resistance to RRD.Rosa rugosa parentage. Most of the accessions were genotyped using the Axiom WagRhSNP 68K SNP array, and their genetic relationships were assessed using Prevosti’s distance [A total of 108 rose accessions including rose species and a wide array of cultivars  were pladistance , a measudistance . Using Pdistance  and visudistance .The Tennessee (Lily loam) and the Delaware (Elsinboro silt/loam) sites are in the USDA Hardiness zones 6b and 7a, respectively. The fields were organized using a randomized block design with three replications. Each rose plant was an experimental unit. In Tennessee, roses were planted 2.4 m apart in a staggered double rose configuration that was 1.8 m apart. Roses were mulched ~8 cm deep and irrigated with drip irrigation. Irrigation was run to ensure that the plants received one inch of water per week. Fertilization was conducted according to soil test data received each year from the UT Soil and Plant Disease/Pest Diagnostic Center . In Delaware, the rows were spaced 2.4 m apart. Within rows, roses were planted with 0.9 m spacing between each plant. Supplemental watering was provided to the rose, initially by hand and later via drip irrigation. The irrigation was run up to twice a week depending on the rainfall, temperature, and plant age. Roses were planted by hand and kept weeded, mulched, and pruned during the growing season to encourage the growth of soft tissue, which is required by the eriophyid mite vector for feeding and reproduction. No fungicides or insecticides were sprayed during the trials.Rosa multiflora plants infected by the virus and infested by the vector mite (Phyllocoptes fructiphilus), which served as a natural source of the virus and mite. In Tennessee, Knock Out Roses® infected with rose rosette emaravirus (RRV) and infested with the vector mite were dug from a commercial bed in Nashville, TN, and transplanted into the experimental block to enhance disease pressure. The plants in both locations had disease pressure augmented by the release of large numbers of viruliferous mites (600–1200 mites per plant) directly to the roses being screened. This was performed by collecting rosetted shoots from RRV-infected roses and attaching sprigs of these (5–15 cm in length) by clipping with binders or by twist-tying to actively growing shoot tips of a target rose. Augmentation was performed 2–4 times per season with at least one symptomatic shoot attached to each test rose.At both locations, there were Plants were evaluated for RRD symptoms several times from July to October each year. In Tennessee, the plants were rated for the severity of symptom development using a 0 to 3 scale , whereas in Delaware, the plants were rated either with no symptoms or with RRD symptoms. Thus, for analysis, the severity scores were transformed into a binary score for each year and replication. The average score per plant over the 3-year trial was used as the phenotypic score for RRD. Thus, if the plant had RRD symptoms all three years, it had an average score of 1.0, and if the plant was asymptomatic all three years, the average score would be zero.In both locations, plants were tested for the presence of RRV using the RRV2 primer pair . In TennA mixed model was used to analyze the mean RRD evaluations over years with the rose accession as a fixed variable and location as a random variable using JMP Pro 16 software. The calculated means were compared to the overall mean, and the rose accessions were divided into three groups according to rose rosette symptom development—low, moderate, and high. The separation of the low and high groups from the mean score was performed at the 0.10 significance level. The results of virus testing are overlaid on these scores in The spread of the disease throughout the experimental plot as indicated by symptom development was low in the first year (0.08), increased four-fold in the second year to 0.36, and peaked in the third year at 0.47. This general pattern is seen for both locations, although the Delaware location, especially in the first year, had lower levels of symptom development than the Tennessee site. Thus, even with augmentation, it takes a three-year field trial to have good confidence in the plant’s ability to resist the infection by this disease.The interaction between the cultivar and location was significant although the variation explained by the cultivar was much higher. Upon inspection of this interaction, it was clear that the Delaware site did not differentiate among the levels of disease incidence as did the Tennessee location. This is reflected in the greater number of cultivars with low symptom development in Delaware (34 roses) as compared to Tennessee (19 roses). All disagreements among cultivars were explained by the movement of cultivars among adjacent categories . This suggests that the interaction effect is due to non-uniform disease pressure rather than variation in virus pathogenicity.The modern rose is a multi-species complex that was developed initially by combining species from the Gallicanae and Indicae sections, which was followed by using species mainly from the Systylae section and, to a lesser extent, species from the Cinnamomeae and Carolinae sections ,44,45. TR. rugosa  and with R. carolina and R. virginiana (‘Basye’s Blueberry’). The A2 subgroup contains the cultivars with more distant introgressions with these species. As expected, the Canadian cultivars  cluster together within this A2 group. These have parentage derived from R. kordesii, R. arkansana, and R. spinosissima. It is in this A group where most of the roses with no or few symptoms are found.In our genetic relationship tree , the A gR. multifora, R. wichurana, and R. setigera) as well as cultivars with recent introgression from these species. Within this group are two roses that showed no or few symptoms. The predominance of floribundas in group C indicates a significant influence of the Systylae species, especially R. multiflora. There is a lessening of Systylae background in group D with primarily shrub and grandiflora types. Group E contains species from the Indicae section and related cultivars. The hybrid tea types, which consist of intercrosses primarily among species in the Gallicanae and Indicae sections, are within the F and G groups, along with a few floribundas, grandifloras, and climbing shrubs. Group H, which includes cultivars such as Knock Out®, Miracle on the Hudson™, and Carefree Beauty™, are distinct from groups F and G, indicating some introgressions from the section Systylae and perhaps Cinnamomeae. As such, the collection evaluated does represent the wide diversity seen among commercial rose germplasm.The B group contains several species of the Systylae section  and one Asian species (R. rugosa) belong to two closely related sections, Carolinae and Cinnamomeae [R. arkansana, R. blanda, R. carolina, R. californica, and R. palustris) did not show symptom development when they were grafted with infected buds [Among those with no or low symptom development are nine species accessions. Accessions of five North American species  to develop the rose ‘Basye’s Blueberry’ [R. arkansana and R. spinosissima, both of which have been previously reported to be resistant to RRD [R. arkansana as well as through his use of the roses ‘Prairie Princess’ and ‘Assinboine’. ‘Morden Blush’ and ‘Winnipeg Parks’ showed few symptoms, whereas ‘Morden Centennial’ and ‘Morden Fireglow’ showed moderate symptom development. RRV was detected in ‘Winnipeg Parks’ and ‘Morden Centennial’ but not in ‘Morden Blush’ and ‘Morden Fireglow’. These have good fertility with most commercial shrub germplasm and may be good sources of RRD resistance. Currently, there is an experiment with populations developed with ‘Morden Blush’ and ‘Morden Fireglow’ to identify the genes involved in their resistance to RRD.Unfortunately, these have been seldom used in rose breeding due to sexual incompatibility with cultivated roses, with a couple of exceptions. Dr. Robert Basye, in his breeding in Texas, used two species from this group  and R. wichurana (section Systylae). Rosa bracteata, or the McCartney rose, is native to China but, once introduced into the US, became established in the southeastern USA. Rosa bracteata has been reported as resistant to the mite that transmits RRD as the mite was not able to reproduce on it [The other two species accessions with no or low symptoms are ce on it . Unfortuce on it .Rosa wichurana thornless ARE showed few symptoms, whereas R. wichurana poterifolia ARE and R. soulieana RM showed moderate to severe symptoms. In addition, the source of the RRD resistance on LG5 appears to be derived from Rosa wichurana ‘Basye’s Thornless’ [Rosa wichurana [Rosa setigera [Resistance from RRD is found within the Systylae section, which has been used extensively in the commercial breeding of roses ,44,45. Hornless’ ,29. It iichurana . Anothersetigera , which iR. rugosa , Purple Pavement (‘HANpur’), ‘Sir Thomas Lipton’, Star Delight (‘MORstar90’), and ‘Therese Bugnet’). These vary extensively in their sexual compatibility depending on how recently the Rosa rugosa was used in the parentage. Those that are one or two generations from the initial cross with Rosa rugosa, such as ‘Fuzzy Wuzzy’, ‘Purple Pavement’, ‘Sir Thomas Lipton’, ‘Star Delight’, and ‘Therese Bugnet’, have less fertility. Nevertheless, these have some breeding potential in an introgression program to incorporate RRD resistance into a wide diversity of cultivated roses. This is the first report of RRD resistance in this species, although it should be noted that some R. rugosa hybrids, such as ‘Basye’s Purple’, ‘Fru Dagmar Hastrup’, and Linda Campbell (‘MORredrug’), showed moderate symptom development.Another large group of accessions with low symptom development is the hybrids with R. wichurana, the TAMU RRD resistant breeding line M4-4, and the previously reported resistant species R. setigera. Thus, it is possible that these have some Systylae background from which their resistance to RRD is derived.The final two accessions that developed no or low symptoms of RRD over the 3-year trial were the floribunda rose Chuckles (‘SIMmimi’) and the miniature rose Fairy Moss (MORfairpol’). Both of these clustered in the B group, which also contained the resistant accession of rugosa accessions (Rosa rugosa Bailey), R. wichurana thornless ARE, Fair Molly, four Rosa rugosa hybrids , and one Canadian rose (‘Winnipeg Parks’) with Rosa arkansana/spinosissima background. The usefulness of these materials depends on how easily the mite can acquire and transmit the virus from these to a susceptible rose. It is possible that mite populations are lower on these roses because they do not form rosettes in which the Pf populations proliferate, but this remains to be studied experimentally. Lower mite populations could reduce the spread of the vector if, as some believe, the mite is more likely to balloon off to find another host plant under high population conditions .Of those with no or low symptoms, about half tested positive for the presence of RRV. These include one of the All other commercial roses and species roses included in this trial showed moderate to severe symptoms of RRD, and most tested positive for RRV. Those cultivars showing moderate RRD symptoms without a positive RRV diagnosis were ‘Caldwell Pink’, ‘Lafter’, Manetti, ‘Morden Fireglow’, and Sorcerer (‘SAVasorc’). These roses generally developed rosettes of an RRV infection but not until late in the trial. To ensure high virus titer, rosetted tissue was tested at least twice with different samples. The potential reasons for this anomaly are the following. (1.) The rose leaf tissue contains inhibitory phenolic compounds that disrupt the extraction and/or PCR reaction , causingPrevious greenhouse work conducted in Arkansas found that out of the 25 roses examined for their susceptibility using mite and graft inoculations, 24 were susceptible, with 2 of these susceptible roses (Bonica and Homerun) being difficult to infect . Both BoRosa bracteata and only moderate symptom development in Bonica is due to their reported mite resistance. If this translates to lower mite numbers or less feeding, this could be a mechanism of resistance protecting these accessions.Although we have identified roses that did not test positive for RRV and/or develop symptoms of RRD under field conditions, it is not known how the plant is able to resist this viral pathogen or its vector or suppress symptom development. It is possible that the lack of symptom development for It has also been reported that the QTL for partial resistance on LG5 affects both symptom development as well as virus titer as measured by Ct values ,29. ThisRosa wichurana. This species, along with Rosa setigera, was reported to have resistance to RRD, which would support future screening of the R. wichurana- and R. setigera-derived climbing and rambling roses. Many of the roses that developed moderate to high symptoms possess a breeding background containing species primarily in the sections Systylae, Indicae, and Gallicanae.As we examine the genetic background of roses with low RRD symptom development in these trials, common parentage includes species within the Cinnamomeae and Carolinae sections. This is consistent with previous work  and sugg


The water and rapid chloride permeabilities were measured. SEM analysis of the microstructure was also performed. The coarse aggregates used were dolomite (control) and recycled concrete aggregate, incorporating different mineral admixtures, including ground, granulated blast slag, granite, and nanosilica. It was found that the slump values of the dolomite concrete decreased compared with recycled aggregate concrete. Compared to the control mix produced with the recycled aggregate, the slump value of the concrete mixes created with the recycled aggregate increased by approximately 11.1% with the addition of binary cementing materials of 1% NS. The results also indicate that the concrete mix containing the recycled aggregate had the highest compressive strength, tensile strength, and flexural strength compared to that of the dolomite aggregate. Regarding the compressive strength, the addition of 1% NS and 15% slag improved the physico-mechanical properties of the recycled aggregate concretes compared to the other mixes after curing in tap water. Compared to the other mixes, the concrete mix containing 1% NS and 15% slag had a comparatively dense and compact microstructure.In this work, the effects of recycled concrete aggregate, modified with mineral admixtures and nanosilica, on the mechanical properties and performance of concrete after curing in tap water for 28 and 90 days were investigated. The compressive (ƒ Solid waste pollution is currently one of the most pressing issues because of its detrimental effects on the environment, public health, and the global economy. Therefore, it is vital to effectively utilize and recycle various solid wastes for the creation of building materials, etc. ,2. The p2 is completely negligible, and nanosilica promotes the growth of Si chains, thereby forming denser structures. Moreover, nanoscale CNS and C-S-H particles fill the voids between hydrates, thus refining the pore size, increasing the complexity of pores, and improving the microstructure of ITZ, contributing to improved impermeability [According to earlier studies, the addition of industrial wastes like slag, aluminum, and granite to concrete reduces production costs and carbon emissions while also adding new qualities, including improved mechanical capabilities and durability. Many contemporary concrete mixtures are altered using admixtures which enhance the microstructure and lower the concentration of calcium hydroxide produced via cement hydration (CH) by consuming it in a pozzolanic reaction. The fine particle size of the pozzolanic materials found in the concrete mixes creates an abundance of nucleating sites for the precipitation of hydration products. Nanosilica additions improve the mechanical properties (compressive strength) of the resulting cement paste when compared to the addition of microsilica. In the case of nanosilica, the consumption of Ca (OH)eability ,8,9.2) and the calcium hydroxide (CH) created during the cement hydration process. In addition, the refined grains’ physical effects enable dense filling within the cement specimens and decrease the wall effect in the zone where the paste meets the fine and coarse aggregate [Concrete specimens are more homogeneous due to this technique. This results from the reaction between the pozzolanic material’s amorphous silica  and slag (GBFS), combined with recycled aggregate, were studied and evaluated for use in the creation of concrete ,16.Compressive strength, rupture modulus, and shrinkage tests were carried out as part of an experimental program that manufactured concrete using the substitutes of recycled coarse aggregate with ratios of 0%, 50%, and 100%; slag (GBFS) with ratios of 0% and 50%; and fly ash (FA) with ratios 0% and 50%. According to the study’s findings, the addition of more recycled aggregate boosted the concrete’s strength. The quality of the recycled material, as well as the mixing technique, may be related to this performance. High-quality recycled concrete may be produced by mixing recycled coarse aggregates, fly ash, and slag ,18,19.Several studies ,21 examiThe impact of using leftover ceramic aggregate in concrete, both coarse and fine, on the strength of concrete was investigated. According to the findings, the concrete produced using ceramic aggregate waste exhibited an enhanced flexural strength, compressive strength, STS, and elastic modulus. . Equal a2O3) and salts) is composed of a mixture of free metal and non-metal materials. Along with metal oxides produced from the molten alloy, aluminum nitrides and carbides may also be present [Aluminum dross  entered concrete through water or sea winds carrying salt ions that eroded the concrete [The concrete industry can positively impact the environment through the recycling of construction debris, reducing demand for landfill space, conserving resources, saving energy, and reducing pollution, as academics have suggested ,32,33,34concrete ,43,44,45This study thoroughly examined the impacts of recycled aggregates and mineral admixtures  on the physical and mechanical properties of concrete specimens. We utilized typical values for the cement replacement levels in concrete mixes: 15% slag, 10% granite, 1.5% aluminum waste, and 1% nanosilica. The aim of this research was to produce sustainable concrete and lower carbon emissions using local waste materials in Egypt, such as slag, granite powder, nanosilica, and aluminum waste as a replacement for cement and recycled aggregate from old concrete as a replacement for coarse aggregate (dolomite).−), as a contaminant, served as the fine aggregate. Portland cement CEM1-42.5 N, supplied by the Suez cement business in Suez, Egypt, was used in this work. The used cement met the Egyptian Standard Specifications (4756-1/2007). The mechanical and physical properties of the used cement were confirmed through laboratory testing at the Housing and Building National Research Center (as per E.S.S No. 2421/2005), which indicated that it was suitable for concrete work. The measured properties of the cement used in this study are presented in (1)(Egyptian Standard Specification No. 1109 2002).(2)(Egyptian Code of Practice for Reinforced Concrete No. 203 2017).(3)(ASTM C637 2009).A 180 kg pan mixer was used to combine the aggregates, binder material, superplasticizer, and water in order to create concrete. The mixing process involved combining the aggregates for approximately 2 min, adding half the water volume in 15 s, combining the ingredients for 3 min, distributing the binder material and superplasticizer over the aggregate in the pan, and mixing for 30 s. The remaining water was added within the next 30 s after restarting the mixing process. Due to the high surface energy of nanoparticles, they are mixed in a number of stages . After aFresh mixtures were slump-tested for workability, According to ASTM C511, the cubes were cured and subjected to strength tests at 28 and 90 days, as per EN 196-1:2016. A 2500 kN Avery Denison machine, as shown in ASTM C1202 states that a cylindrical specimen of 50 × 100 mm is submerged in a 30% NaCl solution for a chloride permeability test, as shown in Samples from the chosen specimens were taken, processed, and tested under TGA/DTG. Testing was performed on the selected samples of dried paste using a TGA-50 thermal analyzer. The analysis was carried out after completing a compression test at the age of 28 days. A sample of approximately 50 mg (−53 µm) was used with a heating rate of 20 °C/min up to 1000 °C in a dynamic nitrogen atmosphere. The TGA investigations revealed the decomposition of the cement hydrates through diminishing increments on the TGA curves and endothermic peaks on the derivative TGA. The slump test is the simplest way to determine workability. In The tensile strength values of the concrete mixes were evaluated at days 28 and 90. The depth of water penetration under pressure indirectly determines water permeability. According to ASTM C1202-C97, the cylinder specimens underwent RCPT, and after 6 h of applying 60 volts, the Coulombs representing the total charge passed was observed. 2 generation.All chosen samples underwent evaluation at temperatures ranging from 25 °C to 1000 °C. After 28 days of hydration, the TGA/DTG curves of OPC, OPC-1N, OPC-1N-1.5Al, OPC-1N-10G, and OPC-1N-15S were obtained, as displayed in (1)The slump value of dolomite concrete decreases as compared to recycled aggregate concrete. However, the addition of 1% NS binary cementing material increases the slump value of the concrete mix produced with recycled aggregate by approximately 11.1%.(2)The concrete mix with recycled aggregate has higher compressive, tensile, and flexural strength than dolomite aggregate because the recycled aggregate’s physical and mechanical characteristics improve upon interlocking with cement paste.(3)The strength increase is paired with a decrease in chloride penetration, indicating that microcracking affects both chloride penetration and compressive strength.(4)The inclusion of nanosilica and mineral admixtures in recycled aggregate concrete improves its mechanical strength.(5)The micrographs show that concrete mixes with recycled aggregate and mineral admixtures have a more compact microstructure compared to normal dolomite concrete due to an increased C-S-H phase and hydration products.This paper presented an experimental study on the performance of recycled aggregates and mineral admixtures with respect to their mechanical and microstructural properties. The study investigated the materials’ compressive, flexural, and tensile strengths, chloride penetration, and microstructure. The findings provide fundamental knowledge, as follows:


Zea mays PHO1, replacing serine residues with alanine, glutamic acid, and aspartic acid, to discern the effects of phosphorylation. Our findings indicate that phosphorylation exerts no impact on the stability or localization of PHO1. Nonetheless, our enzymatic assays unveiled a crucial role for phosphorylation at the S566 residue within the L80 region of the PHO1 structure, suggesting a potential modulation or enhancement of PHO1 activity. These data advance our understanding of starch biosynthesis regulation and present potential targets for crop yield optimization.Starch phosphorylase (PHO) is a pivotal enzyme within the GT35-glycogen–phosphorylase  superfamily. Despite the ongoing debate surrounding the precise role of PHO1, evidence points to its substantial influence on starch biosynthesis, supported by its gene expression profile and subcellular localization. Key to PHO1 function is the enzymatic regulation via phosphorylation; a myriad of such modification sites has been unveiled in model crops. However, the functional implications of these sites remain to be elucidated. In this study, we utilized site-directed mutagenesis on the phosphorylation sites of  Phosphorylase EC 2.4.1.1), discovered in 1939, is instrumental in the reversible transfer of glucosyl units from glucose-1-phosphate (Glc-1-P) to glycogen, enabling the liberation of inorganic phosphates [.1, discoIpomoea batatas, when its N-terminal amino acid residues are phosphorylated, it serves as an active substrate for the 20S or 26S proteasome [Notably, plant phosphorylases bear structural similarities to animal and bacterial counterparts, like maltodextrin phosphorylase, yet they are distinguished by a unique N-terminus and an additional peptide segment known as the L80 domain ,6,7,8. Toteasome . These iThe expression patterns and subcellular localization of PHO1, particularly within plastids, underscore its significant role in starch biosynthesis, as evidenced by correlations with several starch biosynthetic enzymes ,14. StarZea mays, Oryza sativa, Hordeum vulgare, and Triticum aestivum [Zea mays [The phosphorylation-dependent regulation of major starch biosynthetic enzymes was first elucidated in wheat plants, highlighting the phosphorylation of serine residues in SBEs isoforms  ,23. Phosaestivum ,25,26,27Zea mays . In animZea mays . StructuZea mays .Zea mays PHO1, phosphorylation sites have been identified, including serine 69, threonine 537, serine 547, serine 556, serine 565, and serine 566. To decipher the impact of phosphorylation on PHO1 activity, stability, and subcellular localization, site-directed point mutations were executed at serine 69 and serine 566, and recombinant PHO1 was generated. The findings suggest that phosphorylation at serine 566 may primarily modulate PHO1 activity, while the stability and subcellular localization appear unimpacted by phosphorylation.To elucidate the role of phosphorylation in For the protein expression and purification, the PGEX vector system was employed. The PGEX-6T vector allows for the expression of fusion proteins tagged with Glutathione S-transferase (GST), facilitating easy purification. GST is a widely used tag due to its robust expression and ease of purification using glutathione-based resins. The 651 bp fragment from the C-terminal of PHO1 (PHO1-C) gene was amplified C and cloE. coli BL21 (DE3) strain, which was then induced with 0.5 mmol/L IPTG. After being identified via 10% SDS-PAGE, a significant number of recombinant PHO1 protein bands were obtained from 48 kDa–55 kDa, consistent with the expected theoretical molecular weight  (The constructed prokaryotic expression vector PGEX-6T-PHO1-C was transformed into the am tool) E. The coam tool) G, consisZea mays, differing notably from other model crops. Particularly, PHO1 expression presented a dynamic landscape, being significantly amplified in the endosperm, while diminished in the anthers  analysis, point towards the complex expression pattern of PHO1 in  anthers . IntriguZea mays, observing its presence across all developmental stages from the first DAP onwards. Our data depict a progressive increase in protein expression, reaching its peak between 16 to 22 DAP, before initiating a gradual decline until 26 DAP. However, this decline is punctuated by an unexpected surge in PHO1 expression during the later stages of seed development  reporter gene is a useful tool for determining protein location, by fusing the target gene and the GFP reporter gene. The proteins’ location can be accurately determined by observing the green fluorescence emitted under the laser irradiation of a fluorescence microscope. In this study, a PHO1-eGFP fusion protein vector was constructed (pCAMBIA-2300: PHO1-eGFP) and successfully expressed after transformation. The constructed plasmids containing the mutated PHO1-19t were recombined with the PHO1-eGFP vector to determine the subcellular localization of the mutant proteins. The stability of the mutated PHO1 proteins was not significantly affected. The results showed that the 35S-driven ZmPHO1 and eGFP fusion protein was specifically expressed in the amyloplasts of maize endosperm protoplasts . HoweverE. coli.The expression of the cloned genes was assessed using SDS-PAGE and Western blot techniques. Both non-mutated and mutated ZmPHO1 were expressed by inducing with 1 mM IPTG, as described in the section Materials and Methods. Soluble extracts were collected from the induced and non-induced control groups to analyze the expression of recombinant ZmPHO1. The molecular weight of the ZmPHO1 protein fused with the His label was around 120 kDa. SDS-PAGE electrophoresis revealed clear protein bands between 112 kDa–120 kDa for all recombinant PHO1 fused with the His label . WesternThe activity of PHO1 was determined by measuring the release of Pi from Glc-1-P in the presence of maltodextrins using purified recombinant mutated and non-mutated PHO1. All of the recombinant PHO1, both mutated and non-mutated, were found to have similar activity without any pre-treatment with ATP. However, to investigate the effect of phosphorylation site mutation on the activity of PHO1, recombinant mutated and non-mutated PHO1 were pre-treated with 1 mM ATP to stimulate protein phosphorylation by protein kinases present in the amyloplast lysate before enzyme assays. The results showed that S566 is a critical site for the upregulation of PHO1 activity, as PHO1S566A exhibited significantly lower activity compared to non-mutated PHO1 and PHO1S566E, presumably due to the lack of a phosphorylation site in PHO1S566A. The S69 mutation was also found to affect the activity of PHO1 but to a lesser extent. The results are presented in Zea mays grains at all developmental stages and the lack of PHO1 activity in Zea mays shrunken 4 (Sh4) mutants resulted in reduced starch content [Hordeum vulgare [Zea mays suggests that the expression patterns of SSs, SBEs, DBEs, and DPEs were similar to PHO1, and the activities of these enzymes appear similar to PHO1 as well [Oryza sativa, a mutant lack of PHO1 showed changes in seed size and starch content, highlighting the importance of PHO1 in starch biosynthesis [PHO1 is a major active form of phosphorylase in plants and has a diverse role across different species. In rice, PHO1 accounts for 96% of the total phosphorylase activity and is localized in the amyloplast matrix . Similar content ,37. Simi vulgare . Express as well . In Oryzynthesis . OverallZea mays amyloplast starch biosynthesis.PHO1 plays a crucial role in starch metabolism, particularly in non-photosynthetic cells, by regulating the activities of various enzymes through protein–protein interactions. It has been reported that most isoforms of DBEs, SSs, and DPEs form complexes with PHO1, but the exact fate of these interactions remains unclear. One proposed function of PHO1 in starch biosynthesis is that it acts on malto-oligosaccharides (MOs) to produce linear maltose of sufficient length for subsequent branching reactions by SBEs . MoreovePhosphorylation is a key post-translational modification that regulates the activity and function of many enzymes. In starch biosynthesis, various enzymes including SBEI, SBEIIa, SBEIIb, and PHO1 have been reported to undergo phosphorylation ,33. The According to biochemical evidence, unlike the phosphorylation of PHO1, the available level of Pi may be responsible for determining its activity (phosphorolysis/synthesis) in starch metabolism ,43. The Zea mays were not significantly affected by mutations in the phosphorylation sites, possibly due to the presence of other partners in the plastid. It is observed that the expression patterns of non-mutated PHO1 and two mutated forms were similar, indicating that phosphorylation at these sites may not play a significant role in localization. Interestingly, it was found that dephosphorylation of the S566 and S69 sites may enhance local expression patterns, suggesting a potential role for dephosphorylation in this process. Overall, the study suggests that the phosphorylation sites in PHO1 may not have a significant impact on stability and localization in Zea mays.It is found that the stability and localization of PHO1 in Previous studies have reported the phosphorylation of various PHO1 complexes , but no Zea mays, the phosphorylation of the S566 residue within the L80 region does regulate or amplify the activity of PHO1, emphasizing the significance of this specific phosphorylation site.The transcript levels of PHO1 peak in a pattern that aligns closely with various starch biosynthetic enzymes. Analysis of ZmPHO1 expression suggests its involvement in the synthesis and regulation of maize starch during endosperm development. While the phosphorylation sites do not influence the stability and localization of PHO1, indicating that these sites do not play a role in these aspects for PHO1 in These results highlight the significant role of phosphorylation in modulating the activity of PHO1, a critical enzyme in the early and maturity stages of endosperm development. Therefore, any variations in its activity could have substantial consequences for starch production. Our study offers valuable insights that could enhance our comprehension of starch synthesis, with substantial implications for crop improvement. By generating and studying mutants with augmented or restricted PHO1 activity, our research opens avenues for the development of crop varieties with potentially higher starch content. This could lead to improvements in crop yield and quality. Furthermore, elucidating how the phosphorylation status of PHO1 influences PHO1 activity could pave the way for the creation of crop varieties with altered starch properties.Zea mays seeds was selected to identify the expression level of PHO1 protein and transcripts. The inbred line was grown under normal field conditions at the Chongzhou Research Base of Sichuan Agricultural University, China. Materials were collected at 2-day intervals (from 2 to 30 DAP) and quick-frozen with liquid nitrogen and stored at –80 °C for later use.An Mo17 inbred line of The expression analysis in different developmental stages of seeds was carried out using the expression level data determined by RT-qPCR. The extraction of RNA, cDNA synthesis, and expression analysis was carried out as described by Jian M. . The GAPZea mays seeds according to the protocol described in [The protein was extracted from the developing ribed in . SDS-PAGribed in . After e2+-Phos-tag™ agarose. This mixture was subjected to an incubation process at a temperature of 4 °C for 4 h. Post incubation, the mixture was thoroughly washed thrice with the wash buffer composed of 0.1 M Tris-acetate, and 1 M sodium acetate with pH adjusted to 7.5. Next, an elution buffer containing 0.1 M Tris-acetate, 1 M sodium chloride, and 10 mM sodium phosphate (pH 7.5) was used to elute the enriched phosphoproteins. The elution process involved the application of an alkaline phosphatase reaction solution of 10 μL, which contained 0.5 mM magnesium chloride and 40 µg of protein with 0.002 units of alkaline phosphatase. A control reaction solution of 10 μL, including 0.5 mM magnesium chloride and 40 µg protein, was also prepared. Both solutions were subjected to a temperature of 37 °C for 3 h. Subsequently, Phos-tag™ was employed for further enrichment and elution of the proteins. Lastly, the enriched phosphoproteins were analyzed through SDS-PAGE and immunoblotting techniques to confirm the results.The experiment was performed as described by . Phosphohttps://www.expassy.org/) was used (accessed on 5 December 2020) to predict the phosphorylation sites with default settings.The Expassy online tool (v/v) formic acid. The enrichment of phosphopeptide was conducted employing the High-Select™ Fe-NTA Phosphopeptides Enrichment Kit, as per the guidelines given by the manufacturer . Following lyophilization, the phosphopeptides were suspended once more in 20 µL of loading buffer consisting of 0.1% formic acid. The LC-MS analysis was executed with a timsTOF Pro mass spectrometer  in conjunction with Nanoelute  for 60 min. Peptides were placed onto a C18-reversed phase analytical column , initially with buffer A (0.1% formic acid), and then separated using a linear gradient of buffer B (84% acetonitrile and 0.1% formic acid) at a 300 g/min flow rate. The mass spectrometer was set to operate in positive ion mode, collecting ion mobility MS spectra within the m/z 100–1700 mass range and 1/k0 values varying from 0.6 to 1.6. This was followed by conducting 10 cycles of PASEF MS/MS, with a target intensity of 1.5 k and a threshold of 2500. The active exclusion feature was engaged with a release duration of 0.4 min. The initial data garnered from the mass spectrometry analysis were in the RAW file format, and subsequent library identification and quantitative analysis were conducted with the aid of the MaxQuant software (version 1.5.3.17).The detection of phosphorylated proteins was accomplished through the utilization of the iTRAQ™ technique. This investigation was replicated three times, making use of protein samples from Mo17 maize 25 DAP kernels gathered both during the day and night. The process of sample lysis and protein extraction was executed using SDT buffer, comprising 4% SDS, 100 mM Tris-HCl, and 1 mM DTT at a pH of 7.6. The Bio-Rad BCA Protein Assay Kit  was used to determine the protein concentration. Digestion of the protein was achieved with the help of trypsin, following the filter-aided sample preparation (FASP) procedure. The resulting peptides from each sample were subsequently desalted using Empore™ SPE Cartridges C18, standard density, then concentrated using vacuum centrifugation and reconstituted in a solution containing 0.1% (Zea mays PHO1 (gene of PHO1), after the verification of sequence identity subjected to cloning. The following primers were used to amplify the full length of Zea mays PHO1: Fwd: ATGGCGACGACCACCT; Rev: GAGCACAGAAGAAAGAGCAAAG. The full-length sequence was initially cloned with T-vector (pMD19-T) using the kit from Takara, China, according to the abovementioned instructions and prepared PHO1-19t plasmid. PHO1-19t was used to amplify the full-length Zea mays PHO1 and after the verification of sequence identity was subjected to recombination with pET32a and pCAMBIA-2300-eGFP vectors. The following primers: Fwd: GCCATGGCTGATATCATGGCGACGACCACCTCG CCG; Rev: TTGTCGACGGAGCTCGAATTCGGGAAGGATGGC AGGGCTGAT, having an overhang sequence of pET32a, were used to recombine the full-length Zea mays PHO1 with pET32a vector to produce a PHO1-32a plasmid. The following primers: Fwd: ATTTGGAGAGGACAGGGTACCATGGCGACGACCACCTCG; Rev: GGTACTAGTGTCGACTCT AGACTAGGGAAGGATGGCAGGGC, having an overhang sequence of pCAMBIA-2300-eGFP, were used to recombine the full-length Zea mays PHO1 with pCAMBIA-2300-eGFP to produce a PHO1-2300 plasmid. EcoR I and EcoR V combination of restriction enzymes  was used for the linearization of the pET32a vector. The reaction mixture was prepared as follows: pET32a: 1 μg; EcoR V/EcoR 1: 2 μL; 10 × H Buffer: 2 μL; ddH2O: up to 20 μL. The combination of restriction enzymes Kpn I and Xba I  was used for the linearization of the pCAMBIA-2300-eGFP vector. The reaction mixture was prepared as follows: pCAMBIA-2300-eGFP: 1 μg; Kpn I/Xba I: 2 μL; 10 × H Buffer: 2 μL; ddH2O: up to 20 μL. The reaction mixture was kept at 37 °C for up to 6 h for proper digestion. The product was subjected to running on 1% agarose gel and the linearized vector was purified using the Gel-DNA recovery mini kit . The product was used directly or stored at 4 °C. A Clone-Express II One Step Cloning Kit  was used for recombination cloning. DH5α competent cells were used to transfer and amplify the respective plasmids. The clones that were grown were used for bacteria testing to ensure positive clones. The purified plasmid  was used for sequencing and gene confirmation and kept at 4 °C for further use.Extracted RNA was used to generate cDNA, which was used to amplify the full-length Dpn I. The product was used directly or stored at 4 °C. DH5α competent cells were used to transfer and amplify the respective plasmids. The clones that were grown were used for bacteria testing to ensure positive clones. The purified plasmid  was used for sequencing and gene confirmation and kept at 4 °C for further use.The purified plasmid PHO1-19t was used to mutate the phosphorylation sites serine 69 and serine 566 with base-pair residues which encode alanine, aspartic acid, and glutamic acid instead of serine at their respective position. QuickMutation™ Site-Directed Mutagenesis Kit by Beyotime was used for site-directed point mutations through a PCR reaction. The following primers were used to mutate the serine 69 residue site: Ala-Fwd: TGCAAGGCCCCGTCGCGCCCGCGGAAGGGCTT; Ala-Rev: AAGCCCTTCCGCGGG CGCGACGGGGCCTTGCA; Glu-Fwd: TGCAAGGCCCCGTCGAGCCCGCGGAAGGG CTT; Glu-Rev: AAGCCCTTCCGCGGGCTCGACGGGGCCTTGCA. The following primers were used to mutate the serine 566 residue site: Ala-Fwd: CAGAAGAAGAGAGTGCT GAGGATGAGTTAG; Ala-Rev: CTAACTCATCCTCAGCACTCTCTTCTTCTG; Glu-Fwd: AGAAGAAGAGAGTGAAGAGGATGAGTTAGATCC; Glu-Rev: GGATCTAACTCTTC CTCATCACTCTCTTCTTCT; Asp-Fwd: AGAAGAAGAGAGTGATGAGGATGAGTTAGATCC; Asp-Rev: GGATCTAACTCATCCTCATCACTCTCTTCTTCT. PCR was set up using the synthesized primers  to amplify the complete plasmid following the Beyotime QuickMutation™ Site-Directed Mutagenesis Kit protocol. Following the PCR, the product was further incubated at 37 °C for 5 min after adding the 1 μL of TM His-tag Purification Resin” was used for the purification of the protein by following the steps in the instructions accompanying the product.BL21-receptive cells were used for the expression of protein induced via IPTG treatment. An His-tag protein purification kit bought from Beyotime, China, named “BeyoGold2SO4. After this, 2 mL of 1 N H2SO4 acid solution was added containing 1% ammonium molybdate and 4% ferrous sulfate. After 2 min, the resultant color was read at 700 nm (The activity of PHO1 was determined using the method previously used by . Recombit 700 nm . The expv/v) ratio. Two weeks following the primary immunization, the rabbits received five subsequent subcutaneous injections every week. Each of these booster injections comprised 500 µg of the purified protein combined with 500 µL of Freund’s incomplete adjuvant, maintaining a 1:1 ratio. A week post the final injection, blood was drawn from the rabbit’s carotid artery, and serum was subsequently separated. The serum was treated with the addition of 15% glycerin and 0.2% sodium azide and then stored at a temperature of –80 °C for future use. The performance of the antibody serum was then assessed through a Western blot assay.The polyclonal antibody was generated as described in . SubcutaZea mays seeds were germinated in 1:1 of nutrient soil and zircon for about 3 days to about 1–2 cm in bud length, and transferred to a dark-environment culture at 25 °C for 10 days; the seedlings were exposed for a short time (<12 h) to low light (<378 mol*m−2s−1) before the extraction of the protoplast.Material treatment: g centrifuged for 2 min, the upper clear liquid was removed, 100 μL WI solution was added, this was gently mixed at 23 °C with 2000 Lux light for 12 h to induce the protoplast, and a fluorescent microscope was used to observe the protoplast.The extraction of protoplasts: the seed blades of the treatment were cut into 0.5–1 mm wide blades with razorblade, immersed in an enzymatic solution, placed in a light-absorbing vacuum for 30 min, kept for enzymatic reaction for 3.5 h followed by shaking by hand for 10 min, then the same volume of W5 solution was added to terminate the enzyme reaction, filtered with 75 µm nylon mesh, the filtrate fluid was centrifuged at 80 g for 5 min, precipitation was formed, the upper liquid was removed followed by the slow addition of 5 mL of W5 to re-suspend the pellet, this kept on ice for 30 min, the supernatant liquid was removed, MMG solution was added and the pellet re-suspended so that the final concentration of progenitor was 2 × 105/mL. Then, 20 μg recombinant particles were added to a 2 mL round bottom centrifuge tube bottom and 100 μL native mass was added, gently mixed, 100 µL PEG was added, the bottom of the tube mix was gently beaten, conversion was induced in dark conditions for 15 min, 400 µL W5 solution was added to terminate the reaction, 80× w/v) Cellulase, 0.75% (w/v) Macerozyme, 20 mmol/L MES, 10 mmol/L KCl, 10 mmol/L CaCl2, 1% BSA, 0.6 mol/L mannitol. W5: 154 mmol/L NaCl, 125 mmol/L CaCl2, 5 mmol/L KCl, 2 mmol/L MES-KOH, pH = 5.7. MMG: 4 mmol/L MES, 0.4 mol/L Mannitol, 15 mmol/L MgCl2. PEG-Ca2+: 0.2 mol/L Mannitol, 0.1 mol/L CaCl2, (w/v) 25% PEG 4000. WI: 4 mmol/L MES, 0.5 mol/L Mannitol, 20 mmol/L KCl.Enzymatic solution: 1.5% (


Compressive strength was mainly provided by the strength of the aggregates, with high-strength clay ceramsite having the highest 28-day compressive strength value, followed by ordinary crushed-stone aggregates and lightweight ceramsite. Porosity was mainly influenced by the particle size and shape of the aggregates. Lightweight ceramsite had the highest permeability coefficient among different types of cement-bound aggregates, followed by high-strength clay ceramsite and ordinary crushed-stone aggregates. The quality and compressive strength of pervious concrete specimens decreased with the increase in freeze–thaw cycles; the quality loss was 1.52%, and the compressive strength loss rate was 6.84% after 25 freeze–thaw cycles. Quadratic polynomial regression analysis was used to quantify the relationship of durability and freeze–thaw cycles, with R2 of around 0.98. The results provide valuable insights into the potential applications and benefits of using ceramsite as an aggregate material in pervious concrete for more sustainable and durable infrastructure projects.Concrete with light weight and pervious performance has been widely recognized as an effective and sustainable solution for reducing the negative impacts of urbanization on the environment, as it plays a positive role in urban road drainage, alleviating the urban heat island effect and thermal insulation, as well as seismic performance, etc. This research paper presents a feasibility study of pervious concrete preparation with ceramsite as aggregate. First, pervious concrete specimens with different types of aggregates at various water–cement ratios were prepared, and the mechanical properties of pervious concrete specimens were evaluated based on the compressive strength test. Then, the permeability properties of the pervious concrete specimens with different types of aggregates at various water–cement ratios were characterized. Meanwhile, statistical analysis and regression fitting were conducted. Finally, the analysis of the freeze–thaw durability of pervious concrete specimens with ceramsite as aggregate according to indexes including quality loss rate and strength loss rate was performed. The results show that as the water–cement ratio increased, the compressive strength and permeability coefficient of pervious concrete generally decreased. Compressive strength and permeability coefficient showed a great correlation with the water–cement ratio; the R Concrete is a widely used building material due to its high compressive strength, durability, and low cost ,2,3,4. HResearch on the mechanical properties and permeability of pervious concrete has been carried out in the past and has achieved certain results. The strength of pervious concrete mainly comes from the mutual compression among aggregates and the bonding of cementitious materials ,14. TherThe role of aggregates in concrete’s performance has been studied by many scholars. Aggregates play a crucial role in the performance of concrete; they are granular materials, including coarse and fine aggregates, that directly influence the mechanical properties, durability, and workability of concrete . FirstlyThe increasing demand for concrete has brought new requirements for its performance, while environmental and energy-saving requirements have also put forward higher requirements for concrete. The lightweight nature, composite performance, and environmentally friendly development of concrete structures are important directions . CompareThis feasibility study aimed to investigate the use of ceramsite as an aggregate material in pervious concrete and evaluate its mechanical properties, permeability, and durability. The study involved laboratory experiments to determine the compressive strength, flexural strength, and water-absorption capacity of concrete samples, which allowed us to further explore the influence of the water–cement ratio and aggregate types. The novelty of this study is the statistical analysis of compressive strength, permeability, and frost resistance to quantitatively evaluate the influence of the aggregate type on the performance of pervious concrete. The results of this study provide valuable insights into the potential benefits of and challenges to using ceramsite as an aggregate material in pervious concrete. This information could be used by engineers and construction professionals to design more sustainable and durable infrastructure projects that help mitigate the negative impacts of urbanization on the environment.In this study, as the important components of pervious concrete, the cementitious material used was P.O.42.5 ordinary Portland cement, and the properties of this cement are listed in Aggregates are also among the main materials of pervious concrete and are load-bearing materials. The strength of aggregates largely determines the compressive strength of pervious concrete. Therefore, when selecting aggregates, it is important to consider both the permeability and the strength of the aggregates themselves. Different types of aggregates were selected in this study for pervious concrete: one was ordinary basalt crushed-stone aggregates, with a particle size of 9.5 mm, and the other was ceramsite. Ceramsite aggregates included lightweight ceramsite and high-strength clay ceramsite, as shown in In addition, a common method to improve the strength of pervious concrete is to reduce the water–cement ratio by adding a water-reducing agent (WRA). The main admixtures for pervious concrete in this study were high-efficiency naphthalene-series water-reducing agents, with a water-reducing agent dosage of 3% and a water-reducing rate of 10%.Mg, Mc, and Mw are the amounts of aggregates, cement, and water used per cubic meter of pervious concrete, respectively; rg, rc, and rw are the densities of aggregates, cement, and water, respectively; and p is the porosity of the specimen.Due to the fact that the preparation of pervious concrete does not include the use of fine aggregates such as sand and gravel, it is not possible to use traditional concrete mixing methods in the mix design process of pervious concrete with cement-bonded aggregates. Therefore, this study used the volume filling theory in the Chinese specification “Technical Specification for Pervious Cement Concrete Pavement” (CJJ/T 135-2009)  to deterAccording to the Chinese specification “Technical Specification for Pervious Cement Concrete Pavement” (CJJ/T 135-2009) , the mixAccording to the Chinese specification “Standard for Test Method of Mechanical Properties on Ordinary Concrete” (GB/T 50081-2002) , the comAt present, there are two methods for measuring the permeability coefficient of pervious concrete: one is the constant-head testing method, and the other is the variable-head testing method. In this study, according to the Chinese specification “Technical Specification for Pervious Cement Concrete Pavement” (CJJ/T 135-2009) , the varThe pervious concrete specimens were taken out of the curing box after the curing time of 25 d and immersed in water at 20 °C. The water surface had to be 2 cm to 3 cm above the specimen, so that the pervious concrete specimen was fully saturated with water. Three days later, the specimens were taken out of the water and subjected to a freeze–thaw cycle test on pervious concrete. According to the Chinese specification “Standard for Test Methods of Long-term Performance and Durability of Ordinary Concrete” (GB/T 50082-2009) , the slo2 of the model of around 0.94.The effects of the water–cement ratio on the 28-day compressive strength of pervious concrete are shown in For pervious concrete with different types of cement-bound aggregates, the 28-day compressive strength results of pervious concrete with different types of aggregates obtained with compressive strength tests are shown in 2 of the model of around 0.96.For pervious concrete with different types of cement-bound aggregates, the permeability coefficient results are shown in Freeze–thaw durability, including the quality loss rate and strength loss rate, is plotted in 2 of the model of around 0.98.In We observed the bonding surface and physical properties of high-strength ceramic aggregates and crushed-stone aggregates with hardened cement slurry using an optical microscope, and the observation image results are shown in Based on the above research analysis and discussion, it can be inferred that pervious concrete with ceramsite has advantages such as good permeability, environmental protection, and energy conservation and that it can improve soil permeability. Pervious concrete with ceramsite has good permeability, which can effectively reduce rainwater accumulation and the occurrence of water disasters. It can also reduce rainwater runoff, prevent water pollution and the waste of water resources, and meet the requirements for environmental protection and energy conservation. Additionally, it can improve soil permeability, which is beneficial for plant growth and ecological protection. However, the construction requirements of pervious concrete with ceramsite are high, requiring professional construction technology and equipment, and regular maintenance and cleaning to maintain its permeability, which may increase maintenance costs. Thus, applying the results of this research could provide the basis for the possibility of applications in roads and sidewalks, garages and parking lots, parks and gardens, and casing strings for oil wells.As the water–cement ratio increased, the compressive strength of pervious concrete generally decreased due to higher water content and lower cement ratio in the cement slurry. The thickness and bonding area of the cement slurry layer were directly determined by the aggregate–cement ratio. Compressive strength was mainly provided by the strength of aggregates, with the highest 28-day compressive strength being that of high-strength clay ceramsite, followed by ordinary crushed-stone aggregates and lightweight ceramsite.Pervious concrete’s permeability coefficient decreased with the water–cement ratio due to a decrease in aggregate porosity. Porosity was influenced by aggregate particle size and shape, with larger particles resulting in larger porosity and permeability coefficients. Among different aggregates, lightweight ceramsite had the highest permeability coefficient, followed by high-strength clay ceramsite and ordinary crushed-stone aggregates.The quality of pervious concrete decreased as the freeze–thaw cycles increased, which was caused by internal tension between cementitious material and aggregates. After 25 freeze–thaw cycles, the quality loss was 1.52%. Similarly, compressive strength decreased due to reduced bonding strength between cementitious material and aggregates. After 25 freeze–thaw cycles, the compressive strength loss rate was 6.84%.2 values of approximately 0.94 and 0.9, indicating a good regression fit. Quadratic polynomial regression analysis was employed to assess the relationship of durability and freeze–thaw cycles, yielding an R2 value of around 0.98. After 25 freeze–thaw cycles, pervious concrete specimens exhibited only a small area of detachment of ceramic particles on the surface.Linear regression analysis was used to measure the relationship between the water–cement ratio and the compressive strength or permeability coefficient of pervious concrete. The models showed a strong correlation, with RIn this research paper, a feasibility study of pervious concrete preparation with ceramsite as aggregate is discussed. The mechanical and permeability properties of pervious concrete specimens with different types of aggregates at water–cement ratios were investigated. Meanwhile, statistical analysis and regression fitting were conducted. Finally, the analysis of the freeze–thaw durability of pervious concrete with ceramsite as aggregate according to quality loss rate and strength loss rate was performed. Based on the experimental results, the conclusions were drawn:The results of this research could provide the basis for the possibility of applying pervious concrete in roads and sidewalks, garages and parking lots, parks and gardens, and casing strings for oil wells. This study did not consider the influence of cement slurry’s strength, rheological properties, or aggregate-grading characteristics on the mechanics and permeability of pervious concrete, which could be used as further research directions and potential optimization areas.


The neoadjuvant use of immune checkpoint inhibitor combined with chemotherapy (nICT) or chemoradiotherapy (nICRT) in locally advanced esophageal cancer (EC) is currently an area of active ongoing research. Therefore, we carried out a comprehensive meta-analysis to compare the efficacy and safety of the new strategy with routine neoadjuvant strategy, which included neoadjuvant chemotherapy (nCT) and neoadjuvant chemoradiotherapy (nCRT).MEDLINE (via PubMed), Embase (via OVID), ISI Web of Science database and Cochrane Library were included. And, all of them were searched for eligible studies between January, 2000 and February, 2023. The pathological complete response (pCR) and major pathological response (MPR) were primary outcome of our study. The second outcome of interest was R0 resection rate. Odds ratio (OR) and associated 95% CI were used as the effect indicators comparing the safety and efficiency of the neoadjuvant immunotherapy with the routine neoadjuvant therapy. Fixed-effect model (Inverse Variance) or random-effect model (Mantel-Haenszel method) was performed depending on the statistically heterogeneity.2 = 32.8%, P=0.166). The different results were found in the esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC) subgroups, the estimated OR was 2.35  in the EAC subgroup, and 2.35  in the ESCC subgroup, respectively. The neoadjuvant immunotherapy also showed the advantage in the MPR rates . There was no obvious difference between the neoadjuvant immunotherapy and routine neoadjuvant therapy with respect to surgical resection rate, R0 resection rate, surgical delay rate; while more treatment-related adverse events were observed for the neoadjuvant immunotherapy for pneumonitis/pneumonia  and thyroid dysfunction .There were eight trials with 652 patients were included in our meta-analysis. The estimated pCR rate was higher in the neoadjuvant immunotherapy group  could significantly increase the rates of pCR and MPR, compared with routine neoadjuvant therapy (both nCT and nCRT) in the treatment of locally advanced EC. The neoadjuvant immunotherapy and routine neoadjuvant therapy were with acceptable toxicity. However, randomized studies with larger groups of patients need to performed to confirm these results.https://www.crd.york.ac.uk/prospero/, identifier CRD42020155802. Esophageal cancer is one of the deadliest cancers. As the eighth most commonly diagnosed cancer worldwide, there were 544,000 cancer-related deaths of EC in 2020, ranked sixth of cancer-related mortality . AccordiSurgery remains the mainstay for ESCC or EAC, but surgery alone did not show satisfactory clinical data. Some studies showed that neoadjuvant therapy was the most effective strategy in improving survival of resectable esophageal cancer , 3. At pImmune checkpoint inhibitors (ICIs) combined with chemotherapy, as first line, obviously improved survival data of patients with advanced/metastatic esophageal cancer –11. The We summarized the recent studies and carried out this systematic review and meta-analysis to compare the efficacy and safety of the neoadjuvant immunotherapy with the routine neoadjuvant therapy followed by esophagectomy for patients with locally advanced EC.This study was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) and Meta-analysis of Observational Studies in Epidemiology (MOOSE) reporting guidelines , 19 , Embase (via OVID), ISI Web of Science database and Cochrane Library, between January, 2000 and February, 2023. The language was limited to English. The following search terms or keywords were used: esophageal cancer (MeSH) OR esophageal squamous cell carcinoma OR esophageal adenocarcinoma AND neoadjuvant OR preoperative AND programmed cell death 1 (PD-1) OR programmed cell death ligand 1 (PD-L1) OR immunotherapy (Two authors (H.D.X. and K.L.) independently retrieved the available literature to identify the eligible studies. The studies were chosen on the basis of the following criteria: (a) studies only including patients with esophagus cancer or esophagogastric cancer; (b) the primary efficacy outcomes were pathological complete response rate; complete (R0) tumor resection rate; adverse events of neoadjuvant treat; (c) Randomized Controlled Trials (RCTs) or Retrospective experiments comparing neoadjuvant ICIs combined with nCT or nCRT for treating EC and (d) The experimental design met the requirements and included patients with ESCC and EAC. Exclusion criteria were as the following criteria: (a) studies reporting incomplete or inconsistent outcomes; and (b) duplicate studies, studies reporting animal experiments, case reports, cohort studies, and review articles.Data extraction was respectively and carefully performed by two reviewers (H.D.X. and K.L.). The following information was collected: first author, year of publication, region, characteristics of the study population , TNM stage, treatment therapy, adverse events of neoadjuvant therapies, postoperative complications, and pathological response. If the HR and its 95% CI were not directly provided in the original articles, the extracted survival information and the published risk table were used to reconstruct the survival curve for each included study using the method of David . The extThe neoadjuvant immunotherapy comprised neoadjuvant immune checkpoint inhibitor in combination with chemotherapy (nICT) and neoadjuvant immune checkpoint inhibitor in combination with chemoradiotherapy (nICRT). The routine neoadjuvant therapy included neoadjuvant chemotherapy (nCT) and neoadjuvant chemoradiotherapy (nCRT).The pathological TNM stage was staged according to the 8th edition American Joint Committee on Cancer/Union for International Cancer Control staging system . We usedPathologic complete response (pCR) was defined as no evidence of residual tumor cells of the complete resected tumor specimen of neoadjuvant therapy and resection. The major pathological response (MPR) was defined as less than 10% of residual tumor cells. In the present study, the pCR and MPR rates were considered to be the primary outcomes. R0 resection was defined as a microscopically margin-negative resection without microscopic tumor on the primary tumor bed. The R0 surgical resection rate was set as the secondary outcome for comparing neoadjuvant immunotherapy plus chemotherapy with chemotherapy alone for patients.2 statistics (2 >50%) between studies were found. If low heterogeneity (I2 ≤50%) was between studies fixed-effect model (Inverse Variance) was performed.The primary outcome of interest was pathologic response (pCR and MPR). The second outcome of interest was R0 resection rate. Odds ratio (OR) and associated 95% CI were used as the effect indicators comparing the safety and efficiency of the neoadjuvant immunotherapy with the routine neoadjuvant therapy. To minimize the influence of recall and selection bias that occur in retrospective studies, we performed stratified analyses to assess the association in all cohort studies. The heterogeneity between studies was evaluated with Q and Iatistics . The resSensitivity analysis, subgroup analysis and meta-regression were all performed to explore the sources of heterogeneity. The potential publication bias was further validated by the Egger’s and Begg’s test . All staAfter reviewing 557 publications found using the predefined search terms. All investigators finally agreed to include eight eligible studies –33 with 2 = 32.8%, P=0.166), the data from the subgroups within a single study was pooled using a fixed-effect model. The estimated pCR rate was higher in the neoadjuvant immunotherapy group, including nICT and nICRT  and , respectively. Interestingly, we found the common result  .No difference of R0 resection was founded between two groups . Further analyses of individual grade ≥3 TRAEs showed that the neoadjuvant immunotherapy was associated with more pneumonitis/pneumonia  and surgical delay rate  were comparable between the neoadjuvant immunotherapy and the routine neoadjuvant therapy , and tumor mutation burden (TMB). A recent meta-analysis suggested that tissue-based PD-L1 expression, more than any variable other than dMMR/MSI-H, identified varying degrees of benefit from ICIs-containing therapy . The dMMIncidence rate of TRAEs was higher in the immunotherapy than routine neoadjuvant therapy. Our meta-analysis also suggested the same result, especially in pneumonitis/pneumonia and thyroid dysfunction. Tom van don Ende et al. reported one death due to pneumonitis ; and deaThere were several limitations in our study. Firstly, all included studies were descriptive study and the results have not been evaluated in large-scale controlled trials. Therefore, these findings required further validation by large RCTs. Only the RCTs were the golden standard of comparing the neoadjuvant immunotherapy and the routine neoadjuvant therapy. Secondly, researches for neoadjuvant immunotherapy in EAC remains fairly limited. The few researches were all performed in North America and Europe , 30. TheThe current meta-analysis revealed that the neoadjuvant immunotherapy (nICT and nICRT) could significantly increase the rates of pCR and MPR, compared with routine neoadjuvant therapy (nCT and nCRT) in the treatment of locally advanced EC. The neoadjuvant immunotherapy and routine neoadjuvant therapy were with acceptable toxicity. However, randomized studies with larger groups of patients need to performed to confirm these results.The original contributions presented in the study are included in the article/FL: conceptualization. HX and KL: methodology. HQ and HX: software. YL: formal analysis. HQ, HX and KL: data curation. YZ: writing original draft preparation. YZ, LW, HQ and FY: writing-review and editing. All authors contributed to the article and approved the submitted version.