1.Construction and characterization of recombinant human coagulation factor Ⅶ stable transfected cell lines
Xiaoxiao LI ; Jiabin CHEN ; Jiajun LIU ; Zhifei ZHANG ; Sen ZOU ; Lihua ZHU ; Zhaoyong YANG
Acta Universitatis Medicinalis Anhui 2026;61(1):16-22
ObjectiveTo construct a stable monoclonal human embryonic kidney 293 (HEK293) cell line expressing recombinant human coagulation factor Ⅶ (rhFⅦ) and evaluate the expression level and procoagulant bioactivity of rhFⅦ. MethodsThe plasmid pCDNA3.1-EGFP-FⅦ was transfected into HEK293 cells to verify the effectiveness of the transfection system. The plasmid pCDNA3.1-FⅦ was transfected into HEK293 cells, and monoclonal stable transfected cell lines were selected using geneticin (G418). The transcription of the FⅦ gene was identified by reverse transcription polymerase chain reaction (RT-PCR). The expression level of rhFⅦ in the supernatant of the monoclonal stable transfected cell line was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. The concentration of rhFⅦ was determined by enzyme-linked immunosorbent assay (ELISA), and the procoagulant activity of rhFⅦ was measured by human coagulation factor Ⅶ potency assay. ResultsHEK293 cells transfected with pcDNA3.1-EGFP-FⅦ showed green fluorescence, indicating that rhFⅦ was successfully expressed in the supernatant of HEK293 cells after transient transfection with pcDNA3.1-FⅦ. The monoclonal stable transfected cell line was obtained by G418 screening. RT-PCR identified that the FⅦ gene was integrated into the genome of the monoclonal stable transfected cell line. The cell viability was good as detected by Cell Counting Kit-8, and a single band of rhFⅦ was obtained by purification of the cell supernatant. The highest rhFⅦ expression was (1.27±0.09) mg/L, and the highest procoagulant activity was (380.29±13.80)%. ConclusionThe monoclonal HEK293 cell lines which can express rhFⅦ protein efficiently and stably with excellent procoagulant bioactivity is successfully screened.
2.Validity and reliability of the Chinese version of the Sense of Body Boundaries Survey in college students
Jiajun ZHU ; Yong LIU ; Jianfang ZHAO
Chinese Mental Health Journal 2025;39(2):186-192
Objective:To introduce the Sense of Body Boundaries Survey(SBBS),and examine the validity and reliability of the Chinese version among college students.Methods:A total of 701 college students were sur-veyed to test the construct validity and internal consistency reliability of the Chinese version of the SBBS.A total of 154 college students completed the criterion validity test of the Visual Analogue Scale Assessing Perceived Body Boundaries(VASAPBB),Psychological Boundary Questionnaire(PBQ),Chinese Body Shame Scale(CBSS),Self-liking/Self-competence Scale(SLCS)and Self-Objectification Beliefs and Behaviors Scale(SOBBS),and 117 of them were retested 4 weeks later for the test-retest reliability.Results:The Chinese version of the SBBS was demonstrated bifactor model.General factor was sense of body boundaries,and specific factors were sense of barrier and permeability.The bifactor model fit well with the data(x2/df=3.93,CFI=0.91,TLI=0.91,RMSEA=0.07,AIC=368.05,BIC=482.36).The score of the Chinese version of SBBS was positively correlated with the scores of VASAPBB,PBQ and SLCS(r=0.77,0.66,0.38,Ps<0.001),negatively correlated with CBSS and SOBBS(r=-0.61,-0.56,Ps<0.001).The Cronbach α of the Chinese version of SBBS was 0.94,and the test-retest reli-ability was 0.73.Conclusion:The Chinese version of the Sense of Body Boundaries Survey(SBBS)shows good validity and reliability among college students.
3.Expression of PKMYT1 in glioma and its association with prognosis,drug sensitivity,and immune infiltration
Huiping XIE ; Jiajun LIU ; Jiabin CHEN ; Lihua ZHU ; Zhifei ZHANG ; Zhaoyong YANG
Chinese Journal of Cancer Biotherapy 2025;32(5):498-509
Objective:To analyze by bioinformatics the association between the expression of membrane-associated tyrosine/threonine protein kinase 1(PKMYT1)in glioma with its prognostic value,biological function,drug sensitivity,gene mutation,and immune infiltration.Methods:The differential expression of PKMYT1 was analyzed based on the Chinese Glioma Genome Atlas database(CGGA)and the Cancer Genome Atlas database(TCGA).Pathways likely to be enriched for PKMYT1 were predicted by gene ontology analysis(GO)and gene set enrichment analysis(GSEA).PKMYT1 was subjected to Pearson correlation and gene set variation analysis(GSVA)with cell cycle-related genes and gene sets.The survival prognosis,gene mutation,drug sensitivity and immune infiltration were further analyzed for PKMYT1 high and low expression groups of glioma patients.Results:PKMYT1 was significantly highly expressed in WHO high-grade glioma(P<0.000 1),IDH wild-type glioma(P<0.05),and glioblastoma(P<0.000 1).Overall survival(OS)of patients in the PKMYT1 low expression group was significantly higher than that of the high expression group(P<0.05).Cox regression analysis showed that PKMYT1 expression level was an independent prognostic factor for OS(P<0.05).GO and GSEA analyses showed that the gene sets co-expressed with PKMYT1 were mainly enriched in signaling pathways such as cell cycle,DNA replication and DNA damage repair.Pearson correlation and GSVA analyses showed that the expression of PKMYT1 was significantly and positively correlated with the cell cycle-related genes,gene sets and cell cycle checkpoint genes(P<0.01).Drug sensitivity analysis revealed that patients in the PKMYT1 high expression group had high sensitivity osimertinib,dabrafenib,carmustine and cediranib(P<0.05).Mutation analysis revealed that the IDH1 gene had a higher mutation frequency in the PKMYT1 low expression group.The results of immune infiltration analysis showed that PKMYT1 expression was significantly positively correlated with glioma stroma score(r=0.13,P<0.001),immune score(r=0.11,P<0.01)and ESTIMATE score(r=0.13,P<0.001);and was significantly positively correlated with immune cell infiltration level of regulatory T(Treg)cells and M2-type macrophages(P<0.05).Conclusion:Patients with high PKMYT1 expression have a poorer prognosis,and the mechanism may be related to tumor immune infiltration and cell cycle regulation.PKMYT1 is expected to be a potential target for the diagnosis and treatment of glioma.
4.Exosome in HIV infection:Mechanistic roles and translational potential
Ning DING ; Jiajun LI ; Aiwei ZHU ; Bin SU
Chinese Journal of Experimental and Clinical Virology 2025;39(5):652-659
Exosome play a crucial role in human immunodeficiency virus(HIV)infection. They share high similarities with HIV virions in physicochemical properties and secretion mechanisms:both the formation and secretion of exosome and the budding of HIV depend on the endosomal sorting complex required for transport(ESCRT)complex and VPS4 protein,with their membrane structures being phospholipid bilayers and comparable ranges of diameter and density. These similarities pose challenges to related research. Exosome participate in HIV infection through multiple mechanisms,including promoting HIV entry,inhibiting HIV replication,activating HIV latent reservoirs,and engaging in host antiviral immune responses. In terms of clinical applications,the potential of exosome as biomarkers or vaccine carriers has attracted attention,showing translational prospects. In summary,exosome influence the progression of HIV infection through complex mechanisms,and their application prospects in diagnostic markers,vaccine development,and therapeutic targets merit in-depth exploration.
5.Exosome in HIV infection:Mechanistic roles and translational potential
Ning DING ; Jiajun LI ; Aiwei ZHU ; Bin SU
Chinese Journal of Experimental and Clinical Virology 2025;39(5):652-659
Exosome play a crucial role in human immunodeficiency virus(HIV)infection. They share high similarities with HIV virions in physicochemical properties and secretion mechanisms:both the formation and secretion of exosome and the budding of HIV depend on the endosomal sorting complex required for transport(ESCRT)complex and VPS4 protein,with their membrane structures being phospholipid bilayers and comparable ranges of diameter and density. These similarities pose challenges to related research. Exosome participate in HIV infection through multiple mechanisms,including promoting HIV entry,inhibiting HIV replication,activating HIV latent reservoirs,and engaging in host antiviral immune responses. In terms of clinical applications,the potential of exosome as biomarkers or vaccine carriers has attracted attention,showing translational prospects. In summary,exosome influence the progression of HIV infection through complex mechanisms,and their application prospects in diagnostic markers,vaccine development,and therapeutic targets merit in-depth exploration.
6.The practice and exploration on the continuous medical service in children′s hospital
Jiajun YUAN ; Xiaoli TANG ; Tiantian JING ; Zhenhua ZHU ; Yufen WU ; Xiaowei HU ; Li HONG ; Hao ZHANG
Chinese Journal of Preventive Medicine 2025;59(7):1170-1176
To summarize the clinical practice of continuous medical service for patients at Shanghai Children′s Medical Center, affiliated with Shanghai Jiao Tong University School of Medicine, from September 2023 to December 2024, following the approval of its extended care qualification. This study utilized a mixed-methods research design that integrates quantitative and qualitative approaches. The quantitative study included a total of 117 subjects, with an age range of 18 to 35 years, an average age of 21.56 years, and a median age of 19 years; there were 59 males and 58 females. The disease types covered four major categories: childhood leukemia and solid tumors (68 cases), congenital structural malformations (25 cases), congenital hereditary metabolic diseases (4 cases), and rare diseases (20 cases). Among the subjects, 57.26% (67 cases) were first-time visitors to SCMC. The patients came from 20 provinces, autonomous regions, and municipalities across the country, with 88.03% (103 cases) from outside Shanghai. The treatment outcomes showed improvement or cure in 80.34% (94 cases) of the subjects, and there were no medical complaints. In addition, a qualitative study was conducted to deeply explore the experiences, confusions, and challenges of receiving or implementing continuous medical services from the perspectives of patients and their families, as well as medical staff. According to the inclusion and exclusion criteria, a total of 44 subjects were included in the study, among them, there were 12 patients, 12 family members who were taking care of the patients in SCMC, and 20 corresponding medical staff members. The results of the qualitative study showed that trust in the attending physicians of the children′s specialty hospital, a good doctor-patient relationship, satisfactory treatment outcomes, and support from medical insurance policies are the main driving forces for patients over 18 years old to receive continuous treatment at children′s specialty hospitals. The medical staff of the hospital also believed that this model can promote patient benefits. In conclusion, under the policy support of the Shanghai Municipal Health Commission, the "Six Fixed" Model for continuous treatment established by SCMC has achieved certain positive results in practice. This provides practical references for the development of continuous treatment in China and offers new strategies for the application of preventive medicine in the field of children′s health.
7.Validity and reliability of the Chinese version of the Sense of Body Boundaries Survey in college students
Jiajun ZHU ; Yong LIU ; Jianfang ZHAO
Chinese Mental Health Journal 2025;39(2):186-192
Objective:To introduce the Sense of Body Boundaries Survey(SBBS),and examine the validity and reliability of the Chinese version among college students.Methods:A total of 701 college students were sur-veyed to test the construct validity and internal consistency reliability of the Chinese version of the SBBS.A total of 154 college students completed the criterion validity test of the Visual Analogue Scale Assessing Perceived Body Boundaries(VASAPBB),Psychological Boundary Questionnaire(PBQ),Chinese Body Shame Scale(CBSS),Self-liking/Self-competence Scale(SLCS)and Self-Objectification Beliefs and Behaviors Scale(SOBBS),and 117 of them were retested 4 weeks later for the test-retest reliability.Results:The Chinese version of the SBBS was demonstrated bifactor model.General factor was sense of body boundaries,and specific factors were sense of barrier and permeability.The bifactor model fit well with the data(x2/df=3.93,CFI=0.91,TLI=0.91,RMSEA=0.07,AIC=368.05,BIC=482.36).The score of the Chinese version of SBBS was positively correlated with the scores of VASAPBB,PBQ and SLCS(r=0.77,0.66,0.38,Ps<0.001),negatively correlated with CBSS and SOBBS(r=-0.61,-0.56,Ps<0.001).The Cronbach α of the Chinese version of SBBS was 0.94,and the test-retest reli-ability was 0.73.Conclusion:The Chinese version of the Sense of Body Boundaries Survey(SBBS)shows good validity and reliability among college students.
8.Mechanistic study of Yigan Fupi decoction-mediated repair of the intestinal barrier and reduction of IBS sensitivity through regulation of the PKA/PKC-CREB pathway
Yu CHEN ; Jiajun SHI ; Danting FU ; Qinqin YANG ; Rui FU ; Jiajie ZHU ; Mingjin ZHU ; Xinying LIU ; Mingxian CHEN
Acta Laboratorium Animalis Scientia Sinica 2025;33(4):512-521
Objective To study the therapeutic effects of Yigan Fupi decoction(YGFP)on irritable bowel syndrome(IBS)and its mechanism of action in repairing the intestinal barrier and reducing IBS sensitivity through the PKA/PKC-CREB pathway.Methods Baby rats separated from their mother were randomly divided into a model control(M)and a YGFP group,while baby rats without maternal separation were used as a normal control(N)group.The YGFP group was given YGFP for 4 weeks.Abdominal withdrawal reflux was used to evaluate intestinal sensitivity.Liquid chromatography tandem mass spectrometry and ELISA were used to detect bile acid metabolite concentrations and serum levels of interleukin(IL)-6 and CXCL1,respectively.HE staining was used to observe pathological changes in the colon,and Western Blot and immunohistochemistry were used to analyze the relative protein expression levels of PKA,PKC,CREB,5HT2AR,5-HT7R,ZO-1,and Claudin 1.Results Compared with the normal control group,the M group showed a significantly decreased visceral pain threshold,significantly increased levels of total bile acid metabolites,IL-6,and CXCL1,significantly increased relative expression of PKA,PKC,CREB,5HT2AR,and 5-HT7R,and significantly decreased relative expression of ZO-1 and Claudin 1.Compared with the M group,the YGFP group showed a significantly increased visceral pain threshold,significantly reduced levels of total bile acid metabolites,IL-6,and CXCL1,significantly reduced relative expression of PKA,PKC,CREB,5HT2AR,and 5-HT7R,and increased relative expression of ZO-1 and Claudin 1.Conclusions YGFP effectively improved IBS through a mechanism that may involve repair of the intestinal barrier and reduced sensitivity through the PKA/PKC-CREB pathway.
9.Guideline for Adult Weight Management in China
Weiqing WANG ; Qin WAN ; Jianhua MA ; Guang WANG ; Yufan WANG ; Guixia WANG ; Yongquan SHI ; Tingjun YE ; Xiaoguang SHI ; Jian KUANG ; Bo FENG ; Xiuyan FENG ; Guang NING ; Yiming MU ; Hongyu KUANG ; Xiaoping XING ; Chunli PIAO ; Xingbo CHENG ; Zhifeng CHENG ; Yufang BI ; Yan BI ; Wenshan LYU ; Dalong ZHU ; Cuiyan ZHU ; Wei ZHU ; Fei HUA ; Fei XIANG ; Shuang YAN ; Zilin SUN ; Yadong SUN ; Liqin SUN ; Luying SUN ; Li YAN ; Yanbing LI ; Hong LI ; Shu LI ; Ling LI ; Yiming LI ; Chenzhong LI ; Hua YANG ; Jinkui YANG ; Ling YANG ; Ying YANG ; Tao YANG ; Xiao YANG ; Xinhua XIAO ; Dan WU ; Jinsong KUANG ; Lanjie HE ; Wei GU ; Jie SHEN ; Yongfeng SONG ; Qiao ZHANG ; Hong ZHANG ; Yuwei ZHANG ; Junqing ZHANG ; Xianfeng ZHANG ; Miao ZHANG ; Yifei ZHANG ; Yingli LU ; Hong CHEN ; Li CHEN ; Bing CHEN ; Shihong CHEN ; Guiyan CHEN ; Haibing CHEN ; Lei CHEN ; Yanyan CHEN ; Genben CHEN ; Yikun ZHOU ; Xianghai ZHOU ; Qiang ZHOU ; Jiaqiang ZHOU ; Hongting ZHENG ; Zhongyan SHAN ; Jiajun ZHAO ; Dong ZHAO ; Ji HU ; Jiang HU ; Xinguo HOU ; Bimin SHI ; Tianpei HONG ; Mingxia YUAN ; Weibo XIA ; Xuejiang GU ; Yong XU ; Shuguang PANG ; Tianshu GAO ; Zuhua GAO ; Xiaohui GUO ; Hongyi CAO ; Mingfeng CAO ; Xiaopei CAO ; Jing MA ; Bin LU ; Zhen LIANG ; Jun LIANG ; Min LONG ; Yongde PENG ; Jin LU ; Hongyun LU ; Yan LU ; Chunping ZENG ; Binhong WEN ; Xueyong LOU ; Qingbo GUAN ; Lin LIAO ; Xin LIAO ; Ping XIONG ; Yaoming XUE
Chinese Journal of Endocrinology and Metabolism 2025;41(11):891-907
Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.
10.Mechanistic study of Yigan Fupi decoction-mediated repair of the intestinal barrier and reduction of IBS sensitivity through regulation of the PKA/PKC-CREB pathway
Yu CHEN ; Jiajun SHI ; Danting FU ; Qinqin YANG ; Rui FU ; Jiajie ZHU ; Mingjin ZHU ; Xinying LIU ; Mingxian CHEN
Acta Laboratorium Animalis Scientia Sinica 2025;33(4):512-521
Objective To study the therapeutic effects of Yigan Fupi decoction(YGFP)on irritable bowel syndrome(IBS)and its mechanism of action in repairing the intestinal barrier and reducing IBS sensitivity through the PKA/PKC-CREB pathway.Methods Baby rats separated from their mother were randomly divided into a model control(M)and a YGFP group,while baby rats without maternal separation were used as a normal control(N)group.The YGFP group was given YGFP for 4 weeks.Abdominal withdrawal reflux was used to evaluate intestinal sensitivity.Liquid chromatography tandem mass spectrometry and ELISA were used to detect bile acid metabolite concentrations and serum levels of interleukin(IL)-6 and CXCL1,respectively.HE staining was used to observe pathological changes in the colon,and Western Blot and immunohistochemistry were used to analyze the relative protein expression levels of PKA,PKC,CREB,5HT2AR,5-HT7R,ZO-1,and Claudin 1.Results Compared with the normal control group,the M group showed a significantly decreased visceral pain threshold,significantly increased levels of total bile acid metabolites,IL-6,and CXCL1,significantly increased relative expression of PKA,PKC,CREB,5HT2AR,and 5-HT7R,and significantly decreased relative expression of ZO-1 and Claudin 1.Compared with the M group,the YGFP group showed a significantly increased visceral pain threshold,significantly reduced levels of total bile acid metabolites,IL-6,and CXCL1,significantly reduced relative expression of PKA,PKC,CREB,5HT2AR,and 5-HT7R,and increased relative expression of ZO-1 and Claudin 1.Conclusions YGFP effectively improved IBS through a mechanism that may involve repair of the intestinal barrier and reduced sensitivity through the PKA/PKC-CREB pathway.

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