ObjectiveTransfusion-Related Acute Lung Injury （TRALI） is a common fatal transfusion adverse reaction. Major Histocompatibility Complex （MHC） class I is an important factor involved in the pathogenesis of TRALI； however， the role of complement in itspathogenesis has not been fully elucidated. This study aims to explore the role of complement in MHC class I antibody-mediated TRALI， so as to provide a theoretical basis for clinical prevention and treatment. MethodsThis study established a murine model of transfusion-related acute lung injury （TRALI） based on the "two-hit" theory， with lipopolysaccharide （LPS） as the first hit and MHC class I antibody as the second hit. Male Balb/c mice were randomly divided into seven groups （n=5 per group per experiment）： Naive （blank control）， LPS （first hit only）， Isotype （isotype antibody control）， TRALI （model group）， C5aR1 inhi （C5aR1 antagonist intervention）， C5aR2 inhi （C5aR2 antagonist intervention）， and Anti-C5 （anti-complement C5 antibody intervention）. Rectal temperature was monitored after MHC class I antibody injection. After sample collection， the severity of pulmonary edema was assessed by measuring the lung wet-to-dry weight ratio， histological analysis， and immunohistochemistry. Serum and bronchoalveolar lavage fluid were collected to measure cytokine and complement levels. ResultsMice in the TRALI group exhibited a significant decrease in rectal temperature， an increased lung wet-to-dry weight ratio， elevated serum cytokine levels， and markedly heightened complement C5a levels in bronchoalveolar lavage fluid （P＜0.000 1）. Histopathological examination revealed substantial infiltration of inflammatory cells， predominantly neutrophils accompanied by fewer lymphocytes， plasma cells， and monocytes， along with increased deposition of the membrane attack complex C5b-9 in lung tissues. In contrast， mice treated with anti-C5 antibody demonstrated no significant decrease in rectal temperature. The lung wet-to-dry weight ratio in this group showed no statistical difference compared to either the Naive or Isotype control groups （P＞0.05）. Furthermore， these mice displayed reduced serum cytokine levels， a significant attenuation of inflammatory cell infiltration in the lungs， and a 100% survival rate at the 2-hour time point. However， mice administered either the C5aR1 antagonist or the C5aR2 antagonist failed to be protected and subsequently developed TRALI. ConclusionComplement activation， which forms the membrane attack complex C5b-9， plays a critical role in MHC class I antibody-mediated TRALI. Blocking complement C5 activation can effectively prevent the occurrence of TRALI.

Objective: To explore the role of the c.598G>A mutation of the ITGB3 gene in the occurrence of fetal and neonatal alloimmune thrombocytopenia (FNAIT) through its expression in vitro. Methods: The platelet antibodies in the sera of the affected neonate and her mother were detected using commercial enzyme-linked immunosorbent assay (ELISA), solid-phase agglutination, flow cytometry and the gold standard monoclonal antibody-specific immobilization of platelet antigens (MAIPA). The common human platelet antigen (HPA) genotypes of the neonate and her parents were obtained using the HPA-SSP method. The presence of mutations was analyzed by sequencing the exons of the ITGB3 and ITGA2B genes. The target gene of ITGB3 was obtained by PCR amplification using the existing human platelet cDNA. The wild-type ITGB3 eukaryotic expression vector was constructed by TA cloning technology. The 598G>A mutant ITGB3 eukaryotic expression vector was obtained by point mutation, and the plasmid DNA was co-transfected with that of ITGA2B (αⅡb) into HEK293 cells. The transfected cells stably expressing GP Ⅱb/Ⅲa were screened and obtained. The expression of GP Ⅱb/Ⅲa in 598G>A mutant transfected cells and the presence of antibodies against this mutation in the serum of mother were detected by flow cytometry and MAIPA. Results: Antibodies against HLA-class Ⅰ and GP Ⅱb/Ⅲa glycoproteins were detected in the serum of the neonate's mother, and subsequent HLA antibody-specific testing confirmed the presence of antibodies against HLA-B 57∶01 and A 02∶05. ITGB3 sequencing showed that the neonate and her father carried the c.598G>A point mutation, which results in the change of glutamate to lysine at position 200. Antibodies against GP Ⅱb/Ⅲa glycoproteins were not detected using constructed c.598G>A mutant transfected cells reacted with the maternal serum. Conclusion: The in vitro expression and analysis of the ITGB3 c.598G>A mutation did not support a role for this mutation in the pathogenesis of FNAIT. The establishment of this method facilitates the discovery of new platelet low-frequency antigens, and provides a theoretical foundation for the detection of antibodies against platelet antigens associated with patients with adverse pregnancy and childbirth histories.

Objective: To explore the role of the c.598G>A mutation of the ITGB3 gene in the occurrence of fetal and neonatal alloimmune thrombocytopenia (FNAIT) through its expression in vitro. Methods: The platelet antibodies in the sera of the affected neonate and her mother were detected using commercial enzyme-linked immunosorbent assay (ELISA), solid-phase agglutination, flow cytometry and the gold standard monoclonal antibody-specific immobilization of platelet antigens (MAIPA). The common human platelet antigen (HPA) genotypes of the neonate and her parents were obtained using the HPA-SSP method. The presence of mutations was analyzed by sequencing the exons of the ITGB3 and ITGA2B genes. The target gene of ITGB3 was obtained by PCR amplification using the existing human platelet cDNA. The wild-type ITGB3 eukaryotic expression vector was constructed by TA cloning technology. The 598G>A mutant ITGB3 eukaryotic expression vector was obtained by point mutation, and the plasmid DNA was co-transfected with that of ITGA2B (αⅡb) into HEK293 cells. The transfected cells stably expressing GP Ⅱb/Ⅲa were screened and obtained. The expression of GP Ⅱb/Ⅲa in 598G>A mutant transfected cells and the presence of antibodies against this mutation in the serum of mother were detected by flow cytometry and MAIPA. Results: Antibodies against HLA-class Ⅰ and GP Ⅱb/Ⅲa glycoproteins were detected in the serum of the neonate's mother, and subsequent HLA antibody-specific testing confirmed the presence of antibodies against HLA-B 57∶01 and A 02∶05. ITGB3 sequencing showed that the neonate and her father carried the c.598G>A point mutation, which results in the change of glutamate to lysine at position 200. Antibodies against GP Ⅱb/Ⅲa glycoproteins were not detected using constructed c.598G>A mutant transfected cells reacted with the maternal serum. Conclusion: The in vitro expression and analysis of the ITGB3 c.598G>A mutation did not support a role for this mutation in the pathogenesis of FNAIT. The establishment of this method facilitates the discovery of new platelet low-frequency antigens, and provides a theoretical foundation for the detection of antibodies against platelet antigens associated with patients with adverse pregnancy and childbirth histories.

Turning to critical illness is a common stage of various diseases and injuries before death. Patients usually have complex health conditions, while the treatment process involves a wide range of content, along with high requirements for doctor′s professionalism and multi-specialty teamwork, as well as a great demand for time-sensitive treatments. However, this is not matched with critical care professionals and the current state of medical care in China. Telemedicine, which shortens the distance of medical professionals and the gap of disease diagnosis and treatments in various regions through electronic information, can effectively solve the current problem. Therefore, there is an urgent need to develop a standardized, high-quality visualization telemedicine round system .Therefore, experts have been organized to search domestic and foreign literature on telemedicine round for critically ill patients and to form this consensus based on clinical experiences so as to further improve the level of critical care treatments in regions.

Turning to critical illness is a common stage of various diseases and injuries before death. Patients usually have complex health conditions, while the treatment process involves a wide range of content, along with high requirements for doctor′s professionalism and multi-specialty teamwork, as well as a great demand for time-sensitive treatments. However, this is not matched with critical care professionals and the current state of medical care in China. Telemedicine, which shortens the distance of medical professionals and the gap of disease diagnosis and treatments in various regions through electronic information, can effectively solve the current problem. Therefore, there is an urgent need to develop a standardized, high-quality visualization telemedicine round system .Therefore, experts have been organized to search domestic and foreign literature on telemedicine round for critically ill patients and to form this consensus based on clinical experiences so as to further improve the level of critical care treatments in regions.

Objective To investigate the impact of statistical uncertainty per control point(SUpCP)on dose calculation on volumetric modulated arc therapy(VMAT)for rectum cancer,and to analyze the accuracy and efficiency of calculation.Methods 19 patients with rectum cancer undergoing radiotherapy were selected.The initial VMAT plans were generated on Monaco TPS using SUpCP=3,then changed SUpCP in the dose calculation process as follow:10 SUpCPs(1～10)for each patient,and totally190 VMAT dose distributions were obtained.For plan evaluation,Dmax,Dmean,D95%,V50,homogeneity index(HI),conformity index(CI)of the planning target volume(PTV),dissymmetric variations of bladder,small intestine and femoral head,and time calculation(Time)were analyzed.Patient specific quality assurance(PSQA),dose deviation of isocenter(ΔDISO)and passing rate of three-dimensional dose distribution(γ33,γ32,γ22)between calculated and delivered radiation doses were measured.Results AsSUpC increased,Dmax and HI of PTV,Dmax of bladder were increased,but D95%and V50 of PTV,Time,γ32 and γ22 were decreased(P<0.05).Dmax and CI of PTV,Dmean of bladder,Dmax and Dmean of small intestine and femoral head,ΔDISO and γ33 showed no statistical significance(P>0.05).When ΔDISO<1%,gamma passing rate>90%for all VMAT plan.When SUpCP<6,Dmax of PTV<110%of the prescribed dose was obtained;while SUpCP>2,time for dose calculation was less than 5 min.Conclusion For VMAT plan of rectum cancer on Monaco TPS using XVMC algorithm,3%～5%of statistical uncertainty per control point for dose calculation,and 3%2 mm or 2%2 mm gamma criteria for three-dimensional dose verification is recommended.This study provides clinical application basis for precise dose calculation of VMAT plan of rectum cancer.

Objective To investigate the expression of targeted regulation of microRNA-146a (miR-146a) on E-box zinc finger protein 1 gene (ZEB1) and its related molecular mechanisms involved in neuroprotection and autophagy inhibition in a rat model of cerebral hemorrhage. Methods Forty 8-week-old male SD rats were randomly divided into four groups: sham operation group, model group, miR-146a overexpression group and miR-146a underexpression group, with 10 rats in each group. Except for the sham operation group, the other three groups were induced to establish an aneurysmal spontaneous intracerebral hemorrhage rat model using type VII collagenase. Rats of four groups were fed a high-salt diet for 6 weeks and euthanized after an additional feeding for 20 weeks. Neurological function [modified neurologic severity score (mNSS)] and brain water content were compared among the four groups. FJC staining was used to detect the number of degenerated neurons; TUNEL staining was used to detect apoptosis; hematoxylin-eosin (HE) staining was used to observe pathological changes in brain tissue; quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-146a and ZEB1 mRNA; Western blot was used to detect the expression of ZEB1, autophagy-related proteins human recombinant autophagy effector protein (Beclin 1) and microtubule-associated protein 1 light chain 3 (LC3). Results Compared to the sham surgery group, the model group showed a significant increase in mNSS score, brain water content, the number of degenerated neurons and cell apoptosis rate, and the expression level of miR-146a was significantly decreased, while the expressions of ZEB1 mRNA, as well as ZEB1, Beclin 1and LC3 proteins, were significantly increased in the model group (P < 0.05). Compared to the model group, the miR-146a overexpression group exhibited a significant decrease in mNSS score, brain water content, the number of degenerated neurons and cell apoptosis rate, and the expression level of miR-146a was significantly increased, while the expressions of ZEB1 mRNA, as well as ZEB1, Beclin 1 and LC3 proteins were significantly reduced in the miR-146a overexpression group (P < 0.05). Compared to the model group, the miR-146a low-expression group showed a further increase in mNSS score, brain water content, the number of degenerated neurons and cell apoptosis rate, and the expression level of miR-146a was further decreased, while the expressions of ZEB1 mRNA, as well as ZEB1, Beclin 1 and LC3 proteins were further elevated in the miR-146a low-expression group (P < 0.05). Conclusion Upregulating miR-146a can exert neuroprotective effects by targeting and inhibiting ZEB1 gene and protein expression, as well as regulating autophagy activity.

Objective:To evaluate the effect of urethral stent tube irrigation in preventing postoperative complications in adolescents with hypospadias.Methods:A retrospective analysis was conducted using the clinical data of adolescents with hypospadias treated at the Department of Urology, Children’s Hospital of Chongqing Medical University, from April 2016 to July 2023. Patients were divided into control group and study group according to whether urethral stent tube was inserted in urethra during operation. All surgeries were performed by the same surgeon. For two days before surgery, patients were instructed to cleanse the surgical site daily with body wash and a 5% povidone-iodine solution. In the control group, patients received postoperative care with an indwelling urinary catheter. In the study group, in addition to the urinary catheter, a urethral stent tube was placed in the posterior urethra, and postoperative urethral irrigation with saline was performed. Postoperative complications, including wound infection, urethral fistula, urethral stricture, and urethral diverticulum, were monitored. Categorical data were expressed as frequencies and percentages and were analyzed using the χ2 test or Monte Carlo-based χ2 test. Continuous data with normal distribution are presented as Mean±SD and were analyzed using the t-test. Non-normally distributed continuous data were expressed as M ( Q1, Q3) and were analyzed using the Wilcoxon rank-sum test. P<0.05 was considered statistically significant. Results:A total of 76 patients were enrolled in the study, with 43 patients in the control group [median age 12.9(12.2, 13.7) years, reconstructed urethral length (3.88±1.86) cm] and 33 patients in the study group [median age 12.5(11.3, 14.1) years, reconstructed urethral length (3.30±1.45)cm]. Postoperatively, 32 patients (74.4%) in the control group experienced complications, including 24 cases (55.8%) of wound infection, 16 cases (37.2%) of urethral fistula, 6 cases (14.0%) of urethral stricture, and 2 cases (4.7%) of diverticulum. In the study group, only 1 patient (3.0%) developed both wound infection and urethral fistula. The incidence of wound infection, urethral fistula, and overall complications was significantly lower in the study group than in the control group (all P<0.01). Conclusion:Postoperative urethral stent irrigation in adolescents with hypospadias significantly reduces the incidence of wound infection and urethral fistula.

Objective:To evaluate the effect of urethral stent tube irrigation in preventing postoperative complications in adolescents with hypospadias.Methods:A retrospective analysis was conducted using the clinical data of adolescents with hypospadias treated at the Department of Urology, Children’s Hospital of Chongqing Medical University, from April 2016 to July 2023. Patients were divided into control group and study group according to whether urethral stent tube was inserted in urethra during operation. All surgeries were performed by the same surgeon. For two days before surgery, patients were instructed to cleanse the surgical site daily with body wash and a 5% povidone-iodine solution. In the control group, patients received postoperative care with an indwelling urinary catheter. In the study group, in addition to the urinary catheter, a urethral stent tube was placed in the posterior urethra, and postoperative urethral irrigation with saline was performed. Postoperative complications, including wound infection, urethral fistula, urethral stricture, and urethral diverticulum, were monitored. Categorical data were expressed as frequencies and percentages and were analyzed using the χ2 test or Monte Carlo-based χ2 test. Continuous data with normal distribution are presented as Mean±SD and were analyzed using the t-test. Non-normally distributed continuous data were expressed as M ( Q1, Q3) and were analyzed using the Wilcoxon rank-sum test. P<0.05 was considered statistically significant. Results:A total of 76 patients were enrolled in the study, with 43 patients in the control group [median age 12.9(12.2, 13.7) years, reconstructed urethral length (3.88±1.86) cm] and 33 patients in the study group [median age 12.5(11.3, 14.1) years, reconstructed urethral length (3.30±1.45)cm]. Postoperatively, 32 patients (74.4%) in the control group experienced complications, including 24 cases (55.8%) of wound infection, 16 cases (37.2%) of urethral fistula, 6 cases (14.0%) of urethral stricture, and 2 cases (4.7%) of diverticulum. In the study group, only 1 patient (3.0%) developed both wound infection and urethral fistula. The incidence of wound infection, urethral fistula, and overall complications was significantly lower in the study group than in the control group (all P<0.01). Conclusion:Postoperative urethral stent irrigation in adolescents with hypospadias significantly reduces the incidence of wound infection and urethral fistula.

Objective: To understand the prevalence rate of pre myopia among preschoolers and its associated factors, so as to provide a basis for the formulation of myopia prevention and control policies. Methods: In May 2022, a tatol of 1 070 preschoolers in the Science City Area of Mianyang City were selected by using a cluster sampling method. The visual acuity of preschool children was obtained through refractive examination, and a questionnaire survey was conducted to investigate associated factors of myopia in the preclinical stage by Chi square test and multivariate Logistic regression analysis. Results: The prevalence rate of pre myopia in preschoolers was 67.01%, including 66.30% for boys and 67.76% for girls. There was no significant difference by sex( χ 2=0.26, P > 0.05). Multivariate Logistic regression analysis showed that at least one parent with myopia, too much near work activities, time spent on outdoor activity <2 h per day, lack of sleep, and screen time ≥1 h per day were positively correlated with the occurrence of pre myopia( OR =1.62, 1.46, 1.46, 1.38, 1.53, P <0.01). Conclusion The prevalence of pre myopia among preschool children in Science City Area is high. It is suggested that parents, kindergartens and relevant departments should pay attention to the visual status of preschoolers, increase outdoor activities, ensure adequate sleep and train preschoolers to develop scientific eye hygiene habits.