OBJECTIVE To explore the effects of trichloroisocyanuric acid(TCCA)on the prolifera-tion of spermatogonia by inducing oxidative stress and ferroptosis.METHODS GC-1 cells were cultured in DMEM-F12 medium,and cell proliferation was plotted according to the growth curve.GC-1 cells were treated with TCCA at concentrations of 0(cell control),97,194,and 387 μmol·L-1 for 24 h.Cell viability was detected using the CCK-8 method,apoptosis cells were stained with Hoechst 33342,cell cycle was examined by PI staining method,RT-qPCR was performed to measure the mRNA expres-sion levels of apoptosis-related genes Bax,Fas,oxidative stress-related genes superoxide dismutase 2(SOD2),glutathione peroxidase 4(GPX4),nuclear factor erythroid 2-related factor 2(Nrf2),solute carrier family 7 member 11(SLC7A11),dihydroorotate dehydrogenase(DHODH),DNA methyltransferase 3A(DNMT3A),and DNA methyltransferase 3L(DNMT3L).The Griess method was used to determine the nitric oxide(NO)content,colorimetric method for the malondialdehyde(MDA)level,DCFH-DA fluores-cence probe method for the reactive oxygen species(ROS)level,DNTB colorimetric method for the reduced glutathione(GSH)content,and WST-8 method for the reduced coenzymeⅡ(NADPH)content.RESULTS Compared with the cell control group,the cell survival rates in the TCCA 194 and 387 μmol·L-1 groups decreased significantly(P<0.01),accompanied by nuclear condensation and fragmentation,a significant increase in apoptosis rate(P<0.01),and cell arrest in the G2/M phase(P<0.05).Additionally,in the TCCA 387 μmol·L-1 group,the levels of NO,MDA and ROS increased(P<0.01),while the levels of GSH and NADPH decreased(P<0.01).Moreover,the mRNA expressions of SOD2,GPX4,Nrf2,SLC7A11,and DHODH decreased(P<0.05,P<0.01),while the expressions of Bax,Fas,DNMT3L,and DNMT3A increased(P<0.05,P<0.01).CONCLUSION TCCA exposure reduces the viability of GC-1 cells,inhibits cell proliferation,induces apoptosis of GC-1 cells.The mechanism may be related to the ability of TCCA to enhance oxidate stress,induce ferroptosis,and interfere with the methylation of GC-1 cells.

BACKGROUND: Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome is a severe congenital disorder characterized by vaginal hypoplasia caused by dysplasia of the Müllerian duct. Patients with MRKH syndrome often require nonsurgical or surgical treatment to achieve satisfactory vaginal length and sexual outcomes. The extracellular matrix has been successfully used for vaginal reconstruction. METHODS: In this study, we developed a new biological material derived from porcine vagina (acellular vaginal matrix, AVM) to reconstruct the vagina in Bama miniature pigs. The histological characteristics and efficacy of acellularization of AVM were evaluated, and AVM was subsequently transplanted into Bama miniature pigs to reconstruct the vaginas. RESULTS: Macroscopic analysis showed that the neovaginas functioned well in all Bama miniature pigs with AVM implants. Histological analysis and electrophysiological evidence indicated that morphological and functional recovery was restored in normal vaginal tissues. Scanning electron microscopy showed that the neovaginas had mucosal folds characteristics of normal vagina. No significant differences were observed in the expression of CK14, HSP47, and a-actin between the neovaginas and normal vaginal tissues. However, the expression of estrogen receptor (ER) was significantly lower in the neovaginas than in normal vaginal tissues. In addition, AVM promoted the expression of b-catenin, c-Myc, and cyclin D1. These results suggest that AVM might promotes vaginal regeneration by activating the b-catenin/cMyc/cyclin D1 pathway. CONCLUSION This study reveals that porcine-derived AVM has potential application for vaginal regeneration.

This study explores the application effect of the case-based teaching method based on Xuexitong learning platform in the online teaching of Digestive System, and analyzes the learner's emotional experience, learning behavior, and learning effect in the case-based online teaching. The results of the study show that the case-based online teaching model based on Xuexitong learning platform improves students' online learning interest, and the students have good emotional experience, high learning enthusiasm, good classroom interaction, enhanced self-learning ability before and after class, and good learning effect. In addition, precise teaching can be used for individual students who are not enthusiastic about online learning.

ObjectiveTo observe the architectural parameter changes of the muscles around the knee in middle-aged and elderly women with early knee osteoarthritis (KOA) by musculoskeletal ultrasound, and to clarify the indicators of muscles that affect early KOA. MethodsFrom January to August, 2022, 20 healthy middle-aged and elderly healthy women (controls) and 20 middle-aged and elderly women with unilateral early KOA (KOA group) were recruited through Beijing Bo'ai Hospital, to measure muscle thickness of the popliteus muscle, the muscle thickness and the pennation angle of the quadriceps, as rectus femoris, vastus intermedius, vastus lateralis, vastus medialis longus and vastus medialis oblique. ResultsCompared with the healthy limb of KOA group, and the controls, the muscle thickness of vastus medialis longus, vastus medialis oblique and popliteus muscle of the affected limb of KOA group became thinner (t > 2.133, P < 0.05); the proportion of thickness of vastus medialis oblique to thickness of vastus lateralis became smaller (t > 3.660, P < 0.05). The pennation angle was smaller in the affected limb of KOA group than in the matched dominant side of the controls (t = 3.101, P < 0.05). Logistic regression analysis showed that the muscle thickness of vastus medialis oblique (OR = 0.235, 95%CI 0.068 to 0.805, P = 0.021) and popliteus muscle (OR = 0.387, 95%CI 0.152 to 0.980, P = 0.045) were related to the onset of early KOA. ConclusionThe thickness of vastus medialis longus, vastus medialis oblique and popliteus muscle decrease, and the balance of the strength of vastus medialis oblique and vastus lateralis weakens in the affected limbs of the middle-aged and elderly women with early KOA. The thickness of vastus medialis oblique and popliteus muscle are protective factors for onset of KOA.

Objective:To study the clinical features of children with pertussis and the risk factors of severe pertussis.Methods:A retrospective analysis was performed based on clinical data and laboratory examination results of hospitalized children with pertussis who admitted to the intensive care unit, respiratory department, and emergency general department at Hunan Children′s Hospital from January 2019 to March 2020.According to the age, the patients were divided into age ≤3 months group( n=58)and age >3 months group( n=64). According to sputum culture, 63 cases were divided into negative sputum culture group and 59 cases were positive sputum culture group.The patients were also divided into vaccinated group( n=19)and unvaccinated group( n=103). Severe disease was seen in 28 cases, and the other 94 cases had the modest disease.The clinical characteristics between two groups were compared, and the risk factors of severe pertussis pneumonia were analyzed. Results:The hospitalization days in age ≤3 months group was higher than that in age >3 months group.It was also found that shortness of breath, apnea, cyanosis after coughing, heart rate decline were more common in age ≤3 months group than those in age >3 months group( P<0.05). The incidences of respiratory failure and heart failure in positive sputum culture group were higher than those in negative sputum culture group.Clinical characteristics such as hospitalization days, hospitalization expenses, peak white blood cell count, peak lymphocyte count, and incidence of bacterial infection were higher in severe pertussis group than those in non-severe pertussis group( P<0.05). Four patients were treated with exchange blood transfusion, and one patient died.Logistic regression analysis revealed that fever, wheezing, cyanosis after coughing and white blood cell count>20×10 9/L were risk factors for severe pertussis.White blood cell count of 20×10 9/L and lymphocyte count of 14×10 9/L had the highest sensitivity and specificity in predicting severe pertussis(0.71, 0.78; 0.54, 0.79). Conclusion:The younger the children are, the more likely they have shortness of breath, apnea, cyanosis, heart rate falls, and the longer the hospital stay.Bacterial infection will aggravate pertussis.Patients with fever, wheezing, cyanosis after coughing, and white blood cell count>20×10 9/L are more likely to develop severe pertussis.The white blood cell count >20×10 9/L and the lymphocyte count >14×10 9/L are associated with severe pertussis.

Objective:To investigate the factors affecting phenotypes in the patients of methylmalonic acidemia combined with homocysteinemia cblC type with MMACHC c. 609G>A homologous variant. Methods:A retrospective study on the clinical manifestations, complications, treatment, and outcome in 164patients of cblC type with MMACHC c. 609G>A homologous variant was conducted.The patients were diagnosed by biochemical and genetic analysisfrom January 1998 to December 2020. Results:Among the 164 patients, 2 cases were prenatally diagnosed and began treatment after birth. They are 3 and 12 years old with normal physical and mental development. Twenty-one cases were diagnosed by newborn screening. Among them, 15 cases had with normal development. They were treated fromthe age of two weeks at the asymptomatic period. Six cases began treatment aged 1 to 3 months after onset. Their development was delayed. One hundred and forty-one cases were clinically diagnosed. Their onset age ranges from a few minutes after birth to 6 years old. 110 cases had early-onset (78.0%). 31 cases had late-onset (22.0%). Five of them died. 24 patients lost to follow-up. Of the 141 clinically diagnosed patients, 130 (92.2%) with psychomotor retardation, 69 (48.9%) with epilepsy, 39 (27.7%) with anemia, 30 (21.3%) had visual impairment, 27 (19.1%) had hydrocephalus, 26 (18.4%) had feeding difficulties, 7 (5.0%) with liver damage, and 5 (3.5%) with metabolic syndrome. The frequency of hydrocephalus and seizures was significantly higher in the early-onset group. The urinary methylmalonic acid increased significantly in the patients with epilepsy. During the long-term follow-up, the level of plasma total homocysteine in the seizure-uncontrolled group was significantly higher than that in the seizure-controlled group, the difference had a statistical significance ( P<0.05). Conclusion:Most of the patients with MMACHC c. 609G>A homozygous variant had early-onset disease, with a high mortality and disability rate. If not treated in time, it will lead to neurological damage, resulting in epilepsy, mental retardation, hydrocephalus, and multiple organ damage. Pre-symptomatic diagnosis and treatment are crucial to prevent irreversible neurological damage. Neonatal screening and prenatal diagnosis are important to improve the outcome of the patients.

Objective:To retrospectively analyze the clinical application of extracorporeal membrane oxygenation (ECMO) in severe adenovirus pneumonia, and to evaluate the application value of ECMO in children with severe adenovirus pneumonia.Methods:Children diagnosed with severe adenovirus pneumonia and intervened with ECMO in the Hunan Children′s Hospital from January 1, 2018 to December 31, 2019 were recruited in this study for analyzing.The gender, age, clinical manifestations, mechanical ventilation duration, ECMO duration, the length of hospital stay, complications and prognosis were collected and analyzed.Results:A total of 4 children were included in the study, involving 2 cases were successfully evacuated from ECMO.Finally, 3 children died, and 1 case survived.Three death cases had a longer than 18 days of duration from the onset to the start with ECMO.Their ventilator assist time before star-ting ECMO was 3-5 days, and ECMO intervention time was longer, with the maximum of 27.5 days.The survived case had an 11-day duration from the onset to the start with ECMO, and the ventilator assisted time and ECMO intervention time were 5 days, and less than 10 days, respectively.Conclusions:ECMO treatment for children with severe adenovirus pneumonia has a low success rate, but it is still the most important way to save children.Early application of ECMO can improve the prognosis of children with severe adenovirus pneumonia.

Objective:To summary the mixed infection as well as clinical characteristics and analyze the risk factors for mixed infection of severe adenovirus pneumonia(SAP) in children.Methods:The clinical data of 114 children with SAP were retrospectively analyzed.Multivariate Logistic regression analysis was performed to assess the risk factors for mixed infection.Results:The incidence age was from 6 months to 2 years(62.5%). High fever(94.7%), cough(98.2%), dyspnea(86.8%) and lethargy(95.6%) were the main symptoms.Laboratory examination showed that children with SAP were prone to increased white blood cell count, C-reactive protein, procalcitonin, aspartate aminotransferase, alanine aminotransferase and CK-MB, as well as decreased proportion of CD3 + , CD4 + , CD8 + , CD4 + /CD8 + and NK cells.The main complications intrapulmonary organ were respiratory failure(80.7%). The main complications extrapulmonary organ were circulatory complications (55.3%). SAP was easily combined with other pathogenic infections.Streptococcus pneumoniae(22.9%)was the most common bacterial pathogen.Respiratory syncytial virus(10.0%)were the most common virus, in addition, mycoplasma pneumoniae(17.1%) was also common.Multivariable Logistic regression analysis showed that the decreasing ratio of CD4 + /CD8 + and NK cells, congenital heart disease and congenital airway dysplasia were the independent risk factors for mixed infection of SAP in children( P<0.05). Conclusion:The SAP patients could easily suffer from mixed infection and high fatality rate.Immune dysregulation is the important risk factors for mixed infection of SAP in children.So immunoregulatory treatment is very important.

Objective:To investigate the relationship of triglyceride (TG), fasting blood glucose (FPG) and triglyceride glucose product index (TyG) with the incidence of hypertension, and provide basic data for the prevention and treatment of hypertension in the population.Methods:A total of 23 581 individuals who met the research criteria in Jinchang cohort were selected as the research subjects, the Cox proportional hazard model was used to analyze the relationship of TG, FPG, and TyG with the risk of hypertension. A stratified analysis was conducted by sex.Results:After adjusting for confounding factors, compared with the normal TG group, the HR(95% CI) of the elevated TG margin group and the elevated group were 1.16 (1.01-1.34) and 1.49 (1.30-1.70), respectively in the total population. Among men, they were 1.13 (1.01-1.27) and 1.17 (1.06-1.30), and among women, they were 1.05 (0.88-1.26) and 1.06 (0.88-1.28). Compared with the normal FPG group, the HR (95% CI) of the FPG-impaired group were 1.29 (1.13-1.48) in the total population, 1.26 (1.08-1.48) in men and 1.59 (1.14-2.21) in women. Taking the lowest quartile array as a reference, the HR (95% CI) of the highest quartile array of TyG was 1.73 (1.45-2.07) in the total population, 1.32 (1.14-1.53) in men and 1.87 (1.37-2.54) in women. TG, FPG had a nonlinear dose-response relationship with the risk of hypertension, while TyG had a linear correlation with the risk of hypertension. Conclusions:Higher TG, FPG, and TyG levels are independent risk factors for the incidence of hypertension. People with higher TG, FPG and TyG are at high risk for hypertension, to which close attention should be paid in the prevention and treatment of hypertension.

Objective:To explore the relationship between lipid indicators and the incidence of diabetes, and to compare the diabetes prediction and identification power of traditional lipid combined lipid indicators, in order to explore the best alternative indicators for identifying and predicting diabetes.Methods:Based on the Jinchang cohort, a nested case-control study was conducted in 1 025 new cases of diabetes after excluding patients with malignant tumor and related endocrine, circulatory system disease, then an age (±2 years), gender matched 1∶1 control group of 1 025 cases was set to analyze the relationship between the incidence of diabetes and lipid parameters.Results:Among the traditional lipid parameters, the fourth quartile of TG, TC, and LDL-C indicated higher risks of developing diabetes, which was 14.00 times (95% CI: 9.73-20.15), 2.15 times (95% CI: 1.65-2.79) and 1.66 times (95% CI: 1.29-2.14) than that of the first quartile, respectively. The risk of developing diabetes indicated by the fourth quartile of HDL-C was 0.21 times than that indicated by the first quartile (95% CI: 0.15-0.28). In the combined lipid parameters, the fourth quartile of TG/HDL-C, TC/HDL-C, LDL-C/HDL-C and non-HDL-C indicated higher risks of developing diabetes, which was 14.86 times (95% CI: 10.35-21.34), 8.12 times (95% CI: 5.94-11.01), 5.85 times (95% CI:4.34-7.88) and 5.20 times (95% CI: 3.85-7.03) than that indicated by the first quartile, respectively. The areas under the ROC curve of TG, TC, HDL-C, LDL-C, TG/HDL-C, TC/HDL-C, LDL-C/HDL-C and non-HDL-C were 0.76 (95% CI: 0.74-0.78), 0.59 (95% CI: 0.57-0.61), 0.67 (95% CI: 0.65-0.69), 0.57 (95% CI: 0.55-0.59), 0.77 (95% CI: 0.75-0.78), 0.73 (95% CI: 0.71-0.75), 0.69 (95% CI: 0.67-0.71) and 0.66 (95% CI: 0.64-0.68), respectively. The optimal diabetes predicting point cuts of TG, TC, HDL-C, LDL-C, TG/HDL-C, TC/HDL-C, LDL-C/HDL-C and non-HDL-C were 1.40, 4.70, 1.28, 3.25, 1.17, 3.43, 2.46, and 3.58 mmol/L, respectively. Conclusions:Lipid metabolic disorder is a risk factor for diabetes. TG and TG/HDL-C are the good lipid metabolism indicators for the prediction of diabetic.