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.

Objective To explore the clinical effect of Busshenqiangji decoction combined with adalimumab(Adam)in patients with ankylosing spondylitis.Methods A total of 114 patients with ankylosing spondylitis were included and randomly divided into the Adam group and the combined group,with 57 cases in each group.Patients in the Adam group were given adalimumab by subcutaneous injection,40mg/time,once every other week.Patients in the combined group were treated with Bushenqiangji decoction(1 dose/day)on the basis of the above,and both groups were treated continuously for 12 weeks.The efficacy,Traditional Chinese Medicine(TCM)syndrome score,spinal function,clinical index,serological index and adverse reactions were evaluated between the two groups.Results The total effective rate was 92.98%(53/57)in the combination group,which was higher than that of the Adam group(75.44%,43/57,P＜0.05).After treatment,the total score of TCM syndromes,Bath Ankylosing Spondylitis Disease Activity Index(BASDAI)and Bath Andylosing Spondylitis Function Index(BASFI)scores were decreased in the combination group,and which were lower than those of the adalimumab group(P＜0.05).After treatment,the finger-to-floor distance and occipital bone-to-wall distance were decreased in the combination group,and which were smaller than those in the Adam group(P＜0.05).The chest range of motion increased,and which was greater than that in the Adam group(P＜0.05).After treatment,serum levels of sclerostin and 25 hydroxyvitamin D were significantly increased in the combination group than those in the Adam group(P＜0.05).There were no significant differences in the total incidence rates of adverse reactions,such as skin rash and gastrointestinal reactions between the two groups[10.53%(6/57)vs.14.04%(8/57),P＞0.05].Conclusion Bushenqiangji decoction combined with adalimumab can play a synergistic role in the treatment of patients with ankylosing spondylitis,significantly improving the symptoms of pain and morning stiffness.The mechanism of action may be related to improving the expression of osteosclerosis protein and the increase of 25-hydroxyvitamin D.

OBJECTIVE To explore the efficacy and safety of sofosbuvir-velpatasvir(SOF/VEL)combined with or without ribavirin(RBV)in treatment of the patients with genotype 3(GT3)chronic hepatitis C(CHC)and liver cirrhosis.METHODS Totally 230 patients with CT3 CHC and liver cirrhosis who were treated in Traditional Chi-nese Hospital of Xinjiang Uygur Autonomous Region,Xinjiang Hetian Specialized Hospital of Infectious Diseases and Xinjiang Manasi County People's Hospital from Jun.2018 to Mar.2023 were recruited as the research sub-jects.The clinical curative effects were observed after the subjects were treated with single SOF-VEL or the com-bination with RBV for 12 to 24 weeks.The indexes including high-sensitivity hepatitis C RNA(HCV RNA),blood routine indexes,liver function indexes and noninvasive diagnosis indexes for liver fibrosis were observed,and the sustained virological response 12 weeks after the treatment(SVR12)was analyzed.RESULTS The mean age of the enrolled patients was(42.31±11.18)years old,the male patients accounted for 66.52％,and there were 137 cases of GT3a and 93 cases of GT3b 93,there were 183 cases of CHC,44 cases of compensated cirrhosis(CC)and 3 cases of decompensated cirrhosis(DCC).There were 189 cases of single HCV infection,33 cases of mixed infections of HCV and HIV,6 cases of mixed infections of HBV/HCV and 2 cases of triple infections of HBV/HCV/HIV.The overall SVR12 of the 230 patients was 99.57％,the SVR12 of the GT3a type patients was 100.00％,the GT3b type patients 98.92％.The SVR12 of the patients with CHC,CC and DCC were 99.45％,100.00％and 100.00％,respectively.The SVR12 of the patients with single HCV infection,HCV/HIV infec-tion,HBV/HCV infection and HBV/HCV/HIV were 99.47％,100.00％,100.00％and 100.00％,respective-ly.No patient quit the direct-acting antivirals(DAAs)treatment due to the drug-induced adverse reactions.1 pa-tient had relapse due to irregular administration of DAAs.CONCLUSION The virological response rate is high a-mong the patients with GT3 CHC and liver cirrhosis who are treated with single SOF/VEL or the combination with RBV,with the safety favorable.

Objective To explore the diagnostic value of 18F-fluorodeoxyglucose(FDG)PET/CT in primary pulmonary enteric adenocarcinoma.Methods The clinical and imaging data of 9 patients with primary pulmonary enteric adenocarcinoma who under-went 18F-FDG PET/CT examination were retrospectively analyzed,including lesion distribution,morphology,maximum standardized uptake value(SUVmax),clinical symptoms and signs,gastroscopy finding,puncture pathological results,and serum tumor markers[carbohydrate antigen 72-4(CA72-4),cytokeratin 19 fragment antigen 21-1(CYFRA21-1),carcinoembryonic antigen(CEA),carbo-hydrate antigen 199(CA199)].Results Pathological examination confirmed a diagnosis of primary pulmonary enteric adenocarcinoma after excluding gastrointestinal primary tumors through clinical evaluation.In all nine patients,18F-FDG PET/CT examination did not reveal any evidence of digestive system malignancies,and gastrointestinal microscopy was negative.Primary lesions were observed as masses or nodular types in 6 cases(5 in the left lung and 1 in the right lung),while 3 cases exhibited diffuse bilateral pulmonary involvement(manifested as multiple patchy opacities,nodules,ground-glass opacities,and consolidations).All pulmonary primary lesions showed increased 18F-FDG uptake,with SUVmax ranging from 2.7 to 12.8,mean 8.6±3.7.The six masses-or nodular-type primary lesions showed maximum diameters ranging from 2.1 to 10.5 cm,mean(5.23±3.06)cm.Four cases demonstrated hilar and mediastinal lymph node metastases,intrapulmonary metastases,and distant metastases,while 1 case showed only distant metastasis.Elevated levels of serum tumor markers were observed as follows:CA72-4 in 7 cases(10-273.3 U/mL),CEA in 7 cases(5-147.4 ng/mL),CA199 in 6 cases(31.22-4 364 U/mL),and CYFRA21-1 in 5 cases(8.31-99.7 ng/mL).Conclusion When pathological biopsy of a pulmonary lesion suggests primary pulmonary enteric adenocarcinoma after excluding gastrointestinal primary tumors,and 18F-FDG PET/CT shows no gastrointestinal masses,this may support the diagnosis of primary pulmonary enteric adenocarcinoma.

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.

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 explore the clinical effect of Busshenqiangji decoction combined with adalimumab(Adam)in patients with ankylosing spondylitis.Methods A total of 114 patients with ankylosing spondylitis were included and randomly divided into the Adam group and the combined group,with 57 cases in each group.Patients in the Adam group were given adalimumab by subcutaneous injection,40mg/time,once every other week.Patients in the combined group were treated with Bushenqiangji decoction(1 dose/day)on the basis of the above,and both groups were treated continuously for 12 weeks.The efficacy,Traditional Chinese Medicine(TCM)syndrome score,spinal function,clinical index,serological index and adverse reactions were evaluated between the two groups.Results The total effective rate was 92.98%(53/57)in the combination group,which was higher than that of the Adam group(75.44%,43/57,P＜0.05).After treatment,the total score of TCM syndromes,Bath Ankylosing Spondylitis Disease Activity Index(BASDAI)and Bath Andylosing Spondylitis Function Index(BASFI)scores were decreased in the combination group,and which were lower than those of the adalimumab group(P＜0.05).After treatment,the finger-to-floor distance and occipital bone-to-wall distance were decreased in the combination group,and which were smaller than those in the Adam group(P＜0.05).The chest range of motion increased,and which was greater than that in the Adam group(P＜0.05).After treatment,serum levels of sclerostin and 25 hydroxyvitamin D were significantly increased in the combination group than those in the Adam group(P＜0.05).There were no significant differences in the total incidence rates of adverse reactions,such as skin rash and gastrointestinal reactions between the two groups[10.53%(6/57)vs.14.04%(8/57),P＞0.05].Conclusion Bushenqiangji decoction combined with adalimumab can play a synergistic role in the treatment of patients with ankylosing spondylitis,significantly improving the symptoms of pain and morning stiffness.The mechanism of action may be related to improving the expression of osteosclerosis protein and the increase of 25-hydroxyvitamin D.

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.

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.