Stability study of umbilical cord mesenchymal stem cells formulation in large-scale production / 药学学报

Umbilical cord mesenchymal stem cells (UC-MSCs) have been widely used in regenerative medicine, but there is limited research on the stability of UC-MSCs formulation during production. This study aims to assess the stability of the cell stock solution and intermediate product throughout the production process, as well as the final product following reconstitution, in order to offer guidance for the manufacturing process and serve as a reference for formulation reconstitution methods. Three batches of cell formulation were produced and stored under low temperature (2-8 ℃) and room temperature (20-26 ℃) during cell stock solution and intermediate product stages. The storage time intervals for cell stock solution were 0, 2, 4, and 6 h, while for intermediate products, the intervals were 0, 1, 2, and 3 h. The evaluation items included visual inspection, viable cell concentration, cell viability, cell surface markers, lymphocyte proliferation inhibition rate, and sterility. Additionally, dilution and culture stability studies were performed after reconstitution of the cell product. The reconstitution diluents included 0.9% sodium chloride injection, 0.9% sodium chloride injection + 1% human serum albumin, and 0.9% sodium chloride injection + 2% human serum albumin, with dilution ratios of 10-fold and 40-fold. The storage time intervals after dilution were 0, 1, 2, 3, and 4 h. The reconstitution culture media included DMEM medium, DMEM + 2% platelet lysate, 0.9% sodium chloride injection, and 0.9% sodium chloride injection + 1% human serum albumin, and the culture duration was 24 h. The evaluation items were viable cell concentration and cell viability. The results showed that the cell stock solution remained stable for up to 6 h under both low temperature (2-8 ℃) and room temperature (20-26 ℃) conditions, while the intermediate product remained stable for up to 3 h under the same conditions. After formulation reconstitution, using sodium chloride injection diluted with 1% or 2% human serum albumin maintained a viability of over 80% within 4 h. It was observed that different dilution factors had an impact on cell viability. After formulation reconstitution, cultivation in medium with 2% platelet lysate resulted in a cell viability of over 80% after 24 h. In conclusion, the stability of cell stock solution within 6 h and intermediate product within 3 h meets the requirements. The addition of 1% or 2% human serum albumin in the reconstitution diluent can better protect the post-reconstitution cell viability.

Effects of orienteering exercises on myopia prevention and control of primary school students / 中国学校卫生

Objective@#To analyze the eye gaze pattern during the orienteering exercise and effects of orienteering exercises on myopia prevention and control in primary school students, so as to provide reference for the effective prevention and control of myopia.@*Methods@#Visual acuity screening was conducted on fifth grade students aged 10-11 years old in Shaanxi Normal University Primary School, and 120 primary school students with abnormal visual acuity were selected and randomly divided into experimental group ( n =60) and control group ( n =60). The experimental group practiced orienteering cross country program on the basis of the control group, and compared the differences in myopia improvement between the two groups through the characteristics of eye changes tested by ETG spectacle type oculomotor.@*Results@#The differences between the experimental group and the control group in naked eye visual acuity, refraction and visual fatigue before the intervention were not statistically significant ( t=0.13, 1.94, 0.70, P > 0.05); after ten weeks of intervention, the naked eye visual acuity, refraction and visual fatigue of the experimental group were better than those of the control group, and the differences were statistically significant ( t=11.28, 15.03, -247.01, P <0.05). During the process of map recognition and scene recognition, the differences in fixation time, number, frequency and saccade distance of the experimental group were statistically significant ( t=-13.62, -7.83, 4.82, 7.15, P <0.05); the students in the experimental group showed alternate eye gaze characteristics in the process of map recognition and scene recognition.@*Conclusion@#During orienteering exercises, map recognition and real scene recognition mobilized eye following, sweeping, gaze, focus and alternate visual tracking, which have an improved effect on myopia prevention and control in primary school students.

Willingness and related factors of COVID-19 vaccination among middle school students in four cities of China / 中国学校卫生

Objective@#This study aims to investigate the willingness and influencing factors of COVID-19 vaccination among Chinese middle school students, and to provide a reference for promoting the implementation of COVID-19 vaccination among middle school students.@*Methods@#An online survey was conducted among middle school students in Beijing, Anqing of Anhui Province, Xi an of Shaanxi Province and Shenzhen of Guangdong Province, and 9 153 participants were enrolled in the present study. Single factor analysis and multinomial Logistic regression was used to determine the related factors of COVID-19 vaccination willingness.@*Results@#The prevalence of vaccine acceptance, vaccine hesitancy and vaccine refusal among middle school students were 60.05%, 31.59% and 8.36%, respectively. The results of Chi square analysis showed there were significant difference of COVID-19 vaccination willingness among sex, school residence, grade, region, vaccination willingness of surrounding classmates, vaccination willingness of others, fear of SARS-CoV-2 and risk of SARS-CoV-2 infection( χ 2=8.43-1 059.43, P <0.05). Multinomial Logistic regression analysis presented that, compared to "vaccine acceptance" group, those girls, non resident in school, living in Beijing or Anqing, disagree or uncertain with influenced by peers, disagree or uncertain with influenced by others, uncertain with fear of SARS-CoV-2, disagree or uncertain with having risk of SARS-CoV-2 infection were more likely to show vaccine hesitancy( OR =1.22,1.21,1.49,1.69,1.75,2.54,1.41,2.32,3.32,1.99,2.38, P <0.05). And those non boarding in school, living in Beijing or Xi an, disagree or uncertainty influenced by peers, disagree or uncertainty influenced by others, disagree with fear of SARS- CoV- 2, disagree or uncertainty in having risk of SARS-CoV-2 infection were more likely to show vaccine refusal ( OR =1.27, 2.29 ,1.46,3.88,2.37,2.31,1.47,2.14,4.24,1.47, P <0.05).@*Conclusion@#The willingness of COVID-19 vaccination among middle school students in four cities is low, and targeted interventions should be advocated to form herd immunity.

Influence of school climate on influenza vaccination among middle school students in four cities in China / 中国学校卫生

Objective@#This study aims to explore the influencing factors of influenza vaccination in middle school students, and to provide a theoretical basis for improving the influenza vaccination rate and inform interventions.@*Methods@#By using mixed research method, a combination of qualitative research and quantitative research, a survey was conducted among 9 145 middle school students selected from Beijing, Anhui, Shaanxi, Guangdong via the online survey using Wenjuanxing software. Multivariate Logistic regression models were used to analyze the influencing factors of influenza vaccination. After a purposeful sampling, 35 middle school students were invited for semi structured interviews and NVivo plus software were used for data analysis.@*Results@#The self reported influenza vaccination rate of the middle school students was 38.2%. No experience of discussing an influenza vaccine with others ( OR=1.37, 95%CI =1.17-1.60) and devaluing the discussion about vaccination ( OR=1.30, 95%CI =1.08-1.57) was negatively associated with vaccination( P <0.05); The head teacher suggestion(OR=0.39, 95%CI =0.32-0.48), peers vaccination( OR= 0.29, 95%CI =0.25-0.33), and awareness of what the vaccine was and being able to explain it clearly( OR=0.68, 95%CI =0.61-0.75), discussing with others( OR=0.76, 95%CI =0.67-0.86), feeling the discussion is valuable( OR=0.66, 95%CI = 0.59- 0.73), being able to positively influence others( OR=0.61, 95%CI =0.55-0.68), being positively influenced by others ( OR= 0.62, 95%CI =0.54-0.71) were more likely to take vaccination via separated models( P <0.05). The interview results showed that the teacher s cognition and attitude towards the influenza vaccine could directly or indirectly affect the students vaccination decision, and the positive school opinion atmosphere may improve students vaccination rate.@*Conclusion@#It is suggested that the schools should increase students vaccination rate through head teachers guidance and organizing students to participate in discussion on influenza topics.

Clinical analysis on risk factors and prognosis of early post-traumatic epilepsy / Análise clínica de fatores de risco e prognóstico da epilepsia pós-traumática precoce

ABSTRACT Objective To analyze the risk factors and prognosis related to early post-traumatic epilepsy (EPTE). Methods One hundred and eighty-six patients with traumatic brain injury were enrolled. Their full clinical data were collected. Single factor analysis and logistic regression analysis of risk factors related to EPTE were performed. The prognosis of patients was determined. Results Single factor analysis showed that there were significant differences of age (p = 0.011), epilepsy history (p < 0.001), injury site (p = 0.004), injury type (p < 0.001) and injury degree (p < 0.001) between the EPTE group (40 patients) and non-EPTE group (146 patients). Logistic regression analysis showed that the injury site, injury type and injury degree were the main risk factors for EPTE. The odds ratio values of injury site, injury type and injury degree were 1.977 (1.473-2.679), 2.096 (1.543-2.842) and 2.376 (1.864-3.609), respectively. The logistic regression equation was P = Exp (-1.473 + 0.698 × injury site + 0.717 × injury type + 0.935 × injury degree). The sensitivity and specificity of injury site, injury type and injury degree for predicting EPTE were 79.2% and 80.5%, 78.9% and 85.7% and 84.2% and 81.0%, respectively. The analysis of prognosis showed that the Glasgow Outcome Scale/Activity of Daily Living Scale scores in the EPTE group were significantly lower than those in non-EPTE group (p < 0.05). Conclusions Injury site, injury type and injury degree are the main risk factors for EPTE. The prognosis of patients with traumatic brain injury can be affected by EPTE.

RESUMO Objetivo Analisar os fatores de risco e prognóstico relacionados à epilepsia pós-traumática precoce (EPTE). Métodos Cento e oitenta e seis pacientes com lesão cerebral traumática foram incluídos. Seus dados clínicos completos foram coletados. A análise fatorial única e a análise de regressão logística dos fatores de risco relacionados à EPTE foram realizadas. O prognóstico dos pacientes foi observado. Resultados A análise fatorial única mostrou que houve diferenças significativas de idade (p = 0,011), história de epilepsia (p < 0,001), local da lesão (p = 0,004), tipo de lesão (p < 0,001) e grau de lesão (p < 0,001) entre o grupo EPTE (40 casos) e o grupo não-EPTE (146 casos), respectivamente. A análise de regressão logística mostrou que o local da lesão, tipo de lesão e grau de lesão foram os principais fatores de risco para EPTE. Os valores de razões de chance do local da lesão, tipo de lesão e grau de lesão foram 1.977 (1.473-2.679), 2.096 (1.543-2.842) e 2.376 (1.864-3.609), respectivamente. A equação de regressão logística foi P = Exp (-1,473 + 0,698 × local de lesão + 0,717 × tipo de lesão + 0,935 × grau de lesão). A sensibilidade e especificidade do local da lesão, tipo de lesão e grau de lesão para a predição da EPTE foram de 79,2% e 80,5%, 78,9% e 85,7% e 84,2% e 81,0%, respectivamente. A análise do prognóstico mostrou que o escore da Escala de Desfechos de Glasgow / Atividade de Vida Diária no grupo EPTE foi significativamente menor do que no grupo não-EPTE (P <0,05). Conclusões O local da lesão, tipo de lesão e grau de lesão são os principais fatores de risco para EPTE. A EPTE pode afetar o prognóstico de pacientes com lesão cerebral traumática.

Association of Increased Urine Brain Derived Neurotrophic Factor with Lower Urinary Tract Symptoms in Men with Benign Prostatic Hyperplasia / 华中科技大学学报（医学）（英德文版）

Urinary brain-derived neurotrophic factor (BDNF),an ubiquitous neurotrophin,was found to rise in patients with benign prostatic hyperplasia (BPH).We hypothesized that the urinary level of BDNF could be a potential biomarker for lower urinary tract symptoms (LUTS) in patients with BPH.Totally,76 patients with BPH-caused LUTS and 32 male control subjects without BPH were enrolled.International Prostate Symptom Score (IPSS) was applied to assess the symptom severity of LUTS.Urodynamic tests were performed for the diagnosis of underlying detrusor overactivity (DO) in the patients with BPH.Urine samples were collected from all subjects.Urinary BDNF levels were measured using enzyme-linked immunosorbent assays and normalized by urinary creatinine (Cr) levels.Seventy-six BPH patients were divided into moderate LUTS group (n=51,7＜IPSS ≤ 20) and severe LUTS group (n=25,IPSS＞20) according to the IPSS.Of the 76 BPH patients,DO was present in 34 (44.7％)according to the urodynamic test.The urinary BDNF/Cr levels were significantly higher in BPH patients with moderate LUTS (8.29±3.635,P＜0.0001) and severe LUTS (11.8±6.44,P＜0.0001) than normal controls (1.71±0.555).Patients with severe LUTS tended to have higher urinary BDNF/Cr levels than patients with moderate LUTS (11.8±6.44 vs.8.29±3.635,P=0.000).The conditions of BPH with LUTS correlated with elevated urinary BDNF levels,and urinary BDNF levels were even higher in BPH-DO patients.The results of this study have provided evidence to suggest that urinary BDNF level test could evaluate the severity of LUTS in BPH patients,and BDNF level can be used as a biornarker for the diagnosis of DO in BPH patients.

Role of nerve growth factor in the differentiation and proliferation of regulatory T cells induced by mesenchymal stem cells / 中华器官移植杂志

Objective To investigate the influence of nerve growth factor (NGF) on the ability of differentiation and proliferation of regulatory T cells (Tregs) induced by mesenchymal stem cells (MSCs).Methods The MSCs were stimulated directly by NGF.IL-10,TGF-β and HLA-G were tested.The expression of CD4 and CD25 was detected by flow cytometry after co-culture.The expression of CD4,CD25 and Foxp3 was detected by flow cytometry after Transwell co-culture.Results As compared with control group,the expression of IL-10,TGF-β and HLA-G in NGF group was increased (P＜0.05 for all).The number of Tregs was increased after the co-culture (P＜0.05).The reduction in IL-10 and TGF-β could block the inducing function of NGF (P＜0.05).Conclusion NGF can enhance the ability of differentiation and proliferation of Tregs induced by MSC,which is possibly associated with the increases in the expression of IL-10 and TGF-β.

Fusion expression of human renal cell carcinoma-associated antigen G250/MN/CA IX in prokaryotic expression system / 南方医科大学学报

<p><b>OBJECTIVE</b>To achieve high expression of human renal cell carcinoma-associated antigen G250 in Escherichia coli.</p><p><b>METHODS</b>The gene fragments encoding the protein obtained by PCR was cloned into prokaryotic expression vector pET32a(+) and expressed in E. coli Rosseta. The immunogenicity of the recombinant protein was evaluated by Western blotting.</p><p><b>RESULTS</b>The plasmid pET32a(+)/G250 was constructed and expressed in E. coli Rosseta successfully. Western blot analysis showed that the recombinant protein could be specifically recognized by monoclonal antibody M75.</p><p><b>CONCLUSION</b>Efficient G250 expression is achieved in prokaryotic expression system, which may facilitate further functional study of the protein and its monoclonal antibody preparation.</p>

The influence of seed maturity and seed coat on the germination of Bupleurum chinense and B. falcatum / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the factors influencing the seed germination of Bupleurum chinense and B. falcatum.</p><p><b>METHOD</b>The seeds of different maturity, and the seeds whose coat were removed or nicked before dipping in water, were germinated in a light growth chamber.</p><p><b>RESULT</b>The different inflorescence level represents different development period. Seeds harvested from cultivation have different maturity. The germination rate of the seeds in period I to III was nearly zero, and it was significant different from period IV to VII. The seed coat is not an obstacle for water absorption or gas exchange, but its removal led to significant increase of seed germination rate. However, to dip the seeds without coat into water for 48 h had no more effect on germination.</p><p><b>CONCLUSION</b>The seed maturity and germination inhibitors in seed coat are two important factor influencing seed germination in B. chinense and B. falcatum.</p>