Objective: To construct gene recombinant expression plasmids of human anti-P antibody with IgG1 and kappa chain based on the human hybridoma cell line. Methods: Starting from the specific RT-PCR products encoding the variable regions of the IgH chain and IgL chain of the anti-P monoclonal cell line, appropriate restriction enzyme digestion sites were introduced at both ends of the VH and VL fragments through nested PCR. The plasmids carrying the antibody constant region and the nested PCR products of VH and VL were ligated by the action of T4 ligase and subsequently transferred into competent E. coli DH5ɑ, and positive clones were selected in the antibiotic resistant LB medium. After sequence confirmation, recombinant plasmids DNA were transfected into HEK293T cells, and the recombinant antibody obtained in the culture supernatant. The characteristics of recombinant expression antibodies were determined by using rapid antibody isotying kit, serological agglutination tests and flow cytometry. Results: Recombinant gene expression vectors, pFUSEss-CHIg-hG1+VH, pFUSE2ss-CLIg-hκ+VL, were successfully constructed. Human IgG1 kappa light chain antibodies were detected in the supernatant of HEK293T cells transient transfected with recombinant plasmids. After the supernatant was ultra-filtered and concentrated, it could cause agglutination reactions with P antigen-positive red blood cells. The mean fluorescence intensity (MFI) of the reaction between recombinant antibodies and antigen-positive red blood cells in flow cytometry experiments was higher than that of antigen-negative red blood cells. Conclusion: The experimental study on the conversion of red blood group antibody types by genetic engineering technology represents a beneficial exploration towards establishing a feasible technical route for the development of genetic recombination and modification of antibodies reagent.

Objective: To explore the relationship between socio ecological risk factors (SERF), physical activity (PA) developmental trajectories, and depressive symptoms among junior and senior high school students in Tianjin, in order to provide theoretical support for comprehensive interventions for adolescent depression. Methods: A longitudinal follow up design was adopted. In September 2022, a baseline survey was conducted using a stratified cluster random sampling method in two junior high schools and two senior high schools in Tianjin, collecting data on students basic information, SERF, PA, and depressive symptoms. Two follow up surveys were conducted in September 2023 and 2024, yielding 588 valid participants. Latent class growth analysis (LCGA) was used to identify PA developmental trajectory categories among junior and senior high school students. Logistic regression was applied to examine the associations between depressive symptoms and SERF as well as PA trajectories. Results: The detection rates of depressive symptoms among Tianjin junior and senior high school students over the three years were 26.53%, 20.24%, and 21.26 %, respectively. Depressive symptoms were positively correlated with SERF ( OR=1.04, 95%CI=1.03-1.05, P <0.05). The highest risks were observed in the individual dimension and family dimension ( OR =1.28, 1.21, both P <0.05). LCGA identified three PA trajectory groups:persistently low level (80.65%), persistently high level (4.58%), and slowly increasing group ( 14.77 %). Multivariate regression analysis showed that compared with the persistently low level PA, the slowly increasing PA significantly reduced the risk of depressive symptoms ( OR=0.44, 95%CI =0.20-0.88), while SERF still increased the risk of depressive symptoms ( OR=1.04, 95%CI =1.03-1.05) (both P <0.05). Conclusion SERF are risk factors for depression symptoms among junior and senior high school students, whereas slowly increasing PA development trajectory demonstrates a protective effect.

ObjectiveTo understand the seropositivity of neutralizing antibodies (NAb) and low-level NAb against SARS-CoV-2 infection in the community residents, and to explore the impact of COVID-19 vaccination and SARS-CoV-2 infection on the levels of NAb in human serum. MethodsOn the ground of surveillance cohort for acute infectious diseases in community populations in Shanghai, a proportional stratified sampling method was used to enroll the subjects at a 20% proportion for each age group (0‒14, 15‒24, 25‒59, and ≥60 years old). Blood samples collection and serum SARS-CoV-2 NAb concentration testing were conducted from March to April 2023. Low-level NAb were defined as below the 25^th percentile of NAb. ResultsA total of 2 230 participants were included, the positive rate of NAb was 97.58%, and the proportion of low-level NAb was 25.02% (558/2 230). Multivariate logistic regression analysis indicated that age, infection history and vaccination status were correlated with low-level NAb (all P<0.05). Individuals aged 60 years and above had the highest risk of low-level NAb. There was a statistically significant interaction between booster vaccination and one single infection (aOR=0.38, 95%CI: 0.19‒0.77). Compared to individuals without vaccination, among individuals infected with SARS-CoV-2 once, both primary immunization (aOR=0.23, 95%CI: 0.16‒0.35) and booster immunization (aOR=0.12, 95%CI: 0.08‒0.17) significantly reduced the risk of low-level NAb; among individuals without infections, only booster immunization (aOR=0.28, 95%CI: 0.14‒0.52) showed a negative correlation with the risk of low-level NAb. ConclusionsThe population aged 60 and above had the highest risk of low-level NAb. Regardless of infection history, a booster immunization could reduce the risk of low-level NAb. It is recommended that eligible individuals , especially the elderly, should get vaccinated in a timely manner to exert the protective role of NAb.

OBJECTIVE: To observe the effect of acupuncture at "pelvic floor six needles" for mild to moderate female stress urinary incontinence (SUI). METHODS: A total of 60 patients with mild to moderate female SUI were randomly divided into an observation group and a control group, 30 cases each group. The control group received Kegel exercise. The observation group received acupuncture at "pelvic floor six needles" on the basis of the treatment as the control group, bilateral Zhongliao (BL33), Zhibian (BL54), Huiyang (BL35), Shuidao (ST28), Dahe (KI12) and Guanyuan (CV4) were selected, once every other day, 3 times a week, 4 weeks as a course of treatment, a total of 2 courses were required. Before treatment and after 4, 8 weeks of treatment, urine leakage in 1 hour, International Consultation on Incontinence questionnaire short form (ICI-Q-SF) score, and incontinence quality of life questionnaire (I-QOL) score were observed in the two groups, and the clinical efficacy was evaluated. RESULTS: After 8 weeks of treatment, urine leakage in 1 hour and ICI-Q-SF scores in both groups were decreased compared with those before treatment (P<0.05), and urine leakage in 1 hour and ICI-Q-SF score in the observation group were lower than those in the control group (P<0.05). After 4, 8 weeks of treatment, I-QOL scores were increased compared with those before treatment in both groups (P<0.05), and the I-QOL scores in the observation group were higher than those in the control group (P<0.01, P<0.001). The total effective rate of the observation group was 93.3% (28/30), which was higher than 73.3% (22/30) in the control group (P<0.05). CONCLUSION Acupuncture at "pelvic floor six needles" could improve the clinical symptoms and quality of life in patients with mild to moderate female SUI to a certain degree.
Humans ; Female ; Acupuncture Therapy/instrumentation* ; Urinary Incontinence, Stress/physiopathology* ; Middle Aged ; Adult ; Exercise Therapy ; Acupuncture Points ; Pelvic Floor/physiopathology* ; Aged ; Treatment Outcome ; Quality of Life

Extensive epigenetic reprogramming involves in memory CD8+ T-cell differentiation. The elaborate epigenetic rewiring underlying the heterogeneous functional states of CD8+ T cells remains hidden. Here, we profile single-cell chromatin accessibility and map enhancer-promoter interactomes to characterize the differentiation trajectory of memory CD8+ T cells. We reveal that under distinct epigenetic regulations, the early activated CD8+ T cells divergently originated for short-lived effector and memory precursor effector cells. We also uncover a defined epigenetic rewiring leading to the conversion from effector memory to central memory cells during memory formation. Additionally, we illustrate chromatin regulatory mechanisms underlying long-lasting versus transient transcription regulation during memory differentiation. Finally, we confirm the essential roles of Sox4 and Nrf2 in developing memory precursor effector and effector memory cells, respectively, and validate cell state-specific enhancers in regulating Il7r using CRISPR-Cas9. Our data pave the way for understanding the mechanism underlying epigenetic memory formation in CD8+ T-cell differentiation.
CD8-Positive T-Lymphocytes/metabolism* ; Cell Differentiation ; Chromatin/immunology* ; Animals ; Mice ; Immunologic Memory ; Epigenesis, Genetic ; SOXC Transcription Factors/immunology* ; NF-E2-Related Factor 2/immunology* ; Mice, Inbred C57BL ; Gene Regulatory Networks ; Enhancer Elements, Genetic

Objective: To understand the subjective and objective comfort evaluations of students from different age groups on desks and chairs, so as to provide reference for standardized allocation and use of desks and chairs. Methods: From January to April 2024, a total of 2 446 students were selected from 26 schools in 13 districts (counties/cities) in Shanghai, Tianjin, and Wuxi from Jiangsu Province by using cluster random method, including students in kindergartens, primary schools, junior high schools,senior high schools, colleges and universities. Standardized procedures were used to measure the height and weight of participants, and the matching desks and chairs models were selected according to the height. The subjective comfort of students on matching desks and chairs was investigated, and their objective comfort was evaluated by using a self designed questionnaire. The χ 2 test was used to analyze the differences of subjective perception and objective evaluation in comfort between different types of desks and chairs. Results: About 84.1% of the students subjectively thought that large desks and small chairs were very comfortable or relatively comfortable, followed by large desks and chairs (75.7%), and the proportion of small desks and chairs was the lowest among the three types (46.2%), and the difference was statistically significant ( χ 2=722.46, P <0.01). The reporting rates of primary school, junior high school and senior high school students who subjectively considered large desks and chairs to be very comfortable/relatively comfortable were higher than that of other types of desks and chairs, and the differences were statistically significant ( χ 2=297.49, 252.82, 343.67, P <0.01). However, there was no significant difference in the subjective comfort evaluation of different types of desks and chairs among kindergarten children ( χ 2=3.21, P >0.05), and 66.3% of the students in colleges and universities felt very comfortable/relatively comfortable when they used the matching standard desks and chairs. The objective evaluation results of the comfort for the three types of desks and chairs were consistent with the subjective evaluation, but the proportions of the objective evaluation as very comfortable/relatively comfortable were higher than that of the subjective evaluation ( χ 2=20.76- 813.47, P <0.01). Conclusions Large desks and chairs, as well as large desks with small chairs are perceived comfortable, while small desks and chairs are perceived less comfortable. It is recommended to match the large desks and chairs or large desks and small chairs that are suitable for them according to the "standard", to promote physical and mental health of students.

Objective: To investigate the differences in the comfort of desks and chairs furniture among students with different body types according to the standard, so as to provide a reference for guiding students with overweight and obesity to choose the correct study furniture and revising the standards. Methods: From January to April 2024, 2 443 students from 26 schools in 13 districts (counties/cities) in Shanghai, Tianjin, and Jiangsu Province were selected by the cluster random sampling method to conduct physical examination. The subjective and objective evaluations of the comfort of height matched desks and chairs were investigated. The students were divided into non overweight, overweight, and obesity groups according to relevant criteria, and stratified analysis was performed. The χ 2 test was used to analyze differences in the comfort evaluations of desks and chairs among students with different body types. Results: Among the 2 443 students surveyed, 16.7% and 12.6% were respectively classified as overweight and obese. All students assigned the highest comfort ratings to large desks and small chairs (84.1%), and consistency was observed between students subjective and objective evaluations. The reporting rate of samll desks and chairs of obesity students subjective evaluation was lower (36.8%) than that of overweight and non overweight/obesity students (52.1%, 48.0%) ( χ 2=14.63, P <0.01). The overweight and obese group of primary school students had a worse evaluation of the comfort of large desks and chairs and small desks and chairs than those of the non overweight and obese groups( χ 2=15.78, 7.63, P <0.05). Among high school students, the overweight and obese group had worse evaluation of the comfort of large desks and chairs, as well as large desks and small chairs, than those of the non-overweight and obese groups( χ 2=9.62, 11.77, P <0.05). The objective evaluations revealed low compliance ratings on the posture of the thighs and calves for naturally forming an angle greater than 90° (55.6%), and headroom height under the table (50.3%) with small desks and chairs ( χ 2=94.05, 166.47, P <0.05). Conclusions Compared with non overweight/obese students, students with overweight and obese students report poor comfort evaluations of height matched desks and chairs. Revision of the standard should consider the body types of students, and evaluations of the comfort of desks and chairs furniture by students with overweight and obesity should be improved.

Objective:To understand the occurrence of different patterns of multimorbidity among children and adolescents aged 9-18 in Tianjin City and analyze the cumulative effects of lifestyle on these patterns of multimorbidity.Methods:From September to November 2022, a stratified cluster random sampling method was used to select students from primary schools, junior high schools, general high schools, and vocational schools in 16 districts of Tianjin to screen for height, weight, blood pressure, distant vision, and diopter. One year later, a follow-up measurement and questionnaire survey were conducted. The log-binomial model was used to analyze the strength of the association between lifestyle factors and different patterns of multimorbidity.Results:The age of 9 488 students was (12.37±2.49) years old, including 4 999 boys and 4 489 girls. The detection rates of three patterns of multimorbidity of overweight obesity and high blood pressure, overweight obesity and myopia, and overweight obesity with high blood pressure and myopia were 6.63%, 9.32%, and 4.21%, respectively. The detection rates of the three types of multimorbidity in boys were higher than those in girls (all P<0.001). The detection rate of overweight obesity and high blood pressure in suburban areas was higher than that in urban areas ( P=0.002). The detection rate of overweight obesity and myopia in suburban areas was lower than that in urban areas ( P=0.034). The detection rate of overweight obesity and myopia among those aged 9-12 years old was higher than other age groups (all P<0.001). The ARR (95% CI) for the association between favorable physical activity and the occurrence of overweight obesity and high blood pressure was 0.79 (0.68-0.92). The ARR (95% CI) for the association between favorable physical activity and the occurrence of overweight obesity with high blood pressure and myopia was 0.82 (0.67-0.99). Compared with children and adolescents with 0-2 favorable lifestyle factors, those with 4-5 favorable lifestyle factors had a lower risk of overweight obesity and high blood pressure ( ARR=0.84, 95% CI: 0.59-0.92). Conclusion:Boys aged 9-18 in Tianjin City are more prone to multimorbidity of overweight obesity and high blood pressure, overweight obesity and myopia, and overweight obesity with high blood pressure and myopia. Children and adolescents with 4-5 favorable lifestyle factors have a reduced risk of occurrence of overweight obesity and high blood pressure. Lifestyle has cumulative effects on multimorbidity of overweight obesity and high blood pressure.

Objective:To investigate the prevalence status and the clinical characteristics of hepatitis D virus (HDV) among patients chronically infected with hepatitis B virus (HBV) in the Xinjiang region.Methods:A cross-sectional study was conducted. Serum samples from 1 830 patients with chronic HBV infection who visited the Traditional Chinese Medicine Hospital of Xinjiang Uygur Autonomous Region from December 2022 to October 2023 were collected. All sera were tested for anti-HDV IgG and IgM. Sera positive for anti-HDV IgG or IgM were selected for HDV RNA detection. HDV RNA-positive sera were sequenced to determine the HDV genotype. Age, gender, HBV course, and anti-HBV treatment status were used as scoring items based on the propensity score matching (PSM) method. Chronic HBV patients with negative anti-HDV were matched in a ratio of 1∶1. The clinical characteristics of anti-HDV -positive-patients were analyzed. The t-test was used for comparison between groups of normally distributed continuous data. The Wilcoxon signed-rank test was used for comparison between groups of skewness distribution. The χ2 test was used for comparison between groups of enumeration data. Results:The positive detection rates of anti-HDV IgG, anti-HDV IgM, and HDV RNA in 1 830 cases with chronic HBV infection were 2.24% (41/1 830), 1.09% (20/1 830), and 1.69% (31/1 830), respectively. All HDV RNA-positive patients had HDV genotype 1. Two anti-HDV-positive patients had negative hepatitis B surface antigen (HBsAg). Gender, age, HBV course, and anti-HBV treatment status had no significant difference. The quantification of HBsAg, liver biochemical indexes (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and total bile acids), the proportion of patients with liver cirrhosis, and alpha-fetoprotein were significantly higher in the anti-HDV-positive group than in those in the anti-HDV-negative group ( P<0.05). Conclusion:The prevalence rate of HDV in chronic HBV-infected patients at a single center in the Xinjiang region was 2.24%, with the primary genotype being 1. Furthermore, overlap infection should be paid attention to because it might aggravate liver damage.

Objective To establish a method for chiral analysis of dextromethorphan and levomethorphan in hair and urine by high performance liquid chromatography-triple quadrupole mass spectrometry.Methods Hair samples were extracted by grinding ultrasound,and urine samples were extracted by acetonitrile.The supernatant were separated using a Astec CHIROBIOTIC? V2 column(25 cm × 4.6 mm,5 μm)with 0.5 ‰ formic acid and 10 mmol/L ammonium acetate-methanol as mobile phases,then analyzed in scheduled-multiple reaction monitoring mode.Results The limits of detection and lower limits of quantitation of dextromethorphan and levomethorphan were 0.001 ng/mg and 0.005 ng/mg in hair samples,while they were 0.050 ng/mL and 0.200 ng/mL in urine samples.In the corresponding concentration range,there were good linearity for chiral isomers with correlation coefficients over 0.999.Besides,the recovery rates,accuracy and precision met analytical requirements,and no significant matrix effect were exhibited.In the actual case samples,dextromethorphan were detected in hair of 12 cases and in the corresponding urine of 3 cases.Conclusion This method is rapid,sensitive and reliable for chiral analysis of dextromethorphan and levomethorphan in hair and urine.