1.Extraction and purification of Jo-1 antigen
Caizhong ZHU ; Hua CHEN ; Xuexin DENG ; Fulin TANG ; Zhijian YAO
Basic & Clinical Medicine 2006;0(10):-
Objective To improve the purifying method of Jo-1 antigen from rabbit thymus used for detection of anti-Jo-1 antibody by dot-blotting immunoassay(DB).Methods The rabbit thymus glands were cut into pieces,homogenized and extracted by PBS.Total protein was precipitated by acetone to get acetone powder(RTAP).The RTAP was solved in PBS and separated by an by anti-Jo-1 IgG affinity column.Results 5~7 g RTAP was obtained from 100g rabbit thymus glands.There was 19%~24% of protein in RTAP.Jo-1 antigen was enriched around 1900 folds through affinity chromatography,with 2.5% recovery of antigenic activity.In this preparation,there were several bands on SDS-PAGE,but only one band about 50 ku,reacted with anti-Jo-1 antisera on immunoblotting.Dot-blotting also showed that the antigen only reacted with Jo-1 antisera.The purified Jo-1 antigen was not stable for long time,but the antigenic activity could maintain for a long time when there was MgCl2 in the solution.Conclusion Affinity chromatography was a simple and easy method for purifying Jo-1 antigen from rabbit thymus.The antigen purified by affinity chromatography could meet the requirement for detecting Jo-1 antibody bydot-blotting.
2.Research on PEG modification of uricase
Lei CAI ; Xiangdong GAO ; Shu ZHU ; Hua WANG ; Wenbing YAO
Journal of China Pharmaceutical University 2008;(6):557-562
To modify uricase with PEG reagent in order to decrease uricase immunogenieity and increase its stability.Methods:The branched PEG of 40 kD was chosen to modify native uricase.The properties of the mod-ified uricase including the stabilities to protease,pH and temperature,in vivo half-life time,as well as the immu-nogenicity were evaluated.The pharmacokinetic profiles of the midofied uricase were studied in mice.Results:It is demonstrated that the conjugation of PEG to lysine residues of Candida utilis uricase resulted in higher tryp-sin resistance.reduced immune response.and prolonged in vivo half-life.PEG modified uricase retained 80% of the enzymatic activity of native uricase.In addition,it was found that half-life in serum of the intravenously injec-ted PEGylated uriease of up to 696 min was longer that that of native uficase of 45 min.Higher plasma drug con-centrations were also reached with dosing of the PEGylated uricase to mice.Furthermore,the binding affinity Was shown to be reduced for the PEG-uricase using ELISA assay.and it was one-eishth that of native uricase.Final-ly,it Was indicated that the PEG uficase induced a delayed immunoresponse in mice following repeated adminis-trations.Conclusion:These findings demonstrate that this chemically modified form of uricase may serve as a potentially effective drug to treat gout patients.
3.Structure,Function and Evolution of Baculoviral Inhibitor of Apoptosis Protein Gene
Rui ZHANG ; Qing YAO ; Jian-Xin PENG ; Hua-Zhu HONG ;
Microbiology 1992;0(01):-
Baculoviral IAP(inhibitor of apoptosis protein) gene was identified firstly in IAP gene family.The structurcal feature of baculoviral IAP genes are characterized BIR and RING domain;Despite similar to P35 in antiapoptotic function,baculovrial IAP and P35 differ in structure and mechnism of action.Phylogenetic analysis of IAP genes and lots of evidence sppport the origin of this viral gene by capture of a host gene early in the evolution of Lepidoptera.
4.Nicotine dependence among rural-to-urban migrant workers in Shanghai
Yao LIU ; Zhigang PAN ; Tianying WANG ; Tianhao WANG ; Yao SHEN ; Jian GONG ; Wei DAI ; Jin ZHOU ; Hua YANG ; Shanzhu ZHU
Chinese Journal of General Practitioners 2014;(6):439-443
Objective To explore the nicotine dependence levels and the influencing factors of dependence among rural-to-urban migrant workers in Shanghai so as to provide scientific rationales for an effective implementation of tobacco control.Methods Multi-stage randomized sampling was used to select a total of 5 856 rural-to-urban migrant workers from 7 districts in Shanghai.Fagerstrom Test for Nicotine Dependence ( FTND) and logistic regression model were used to analyze the association between nicotine dependence and relevant risk factors.Results The current smoking prevalence was 23.7%.The average FTND score for current smokers , high dependence and low dependence were 3.38 ±2.49, 7.00 ±1.07 and 2.23 ±1.71 respectively.The high and low dependence rates were 21.7% and 78.3%.The results of logistic regression showed that education , duration of smoking and length of migration were associated with nicotine dependence.Lower level of education was more prone to high nicotine dependence.Those with a lower education had a higher risk of nicotine dependence ( junior high school: OR=1.742, primary school or lower: OR =1.994 vs.senior high school or higher ).High nicotine dependence increased with the duration of smoking (smoking 6-10 years:OR=3.007, 11-15 years:OR=4.076, 16-20 years:OR=5.451, 20 years or more:OR=5.726 vs.5 years or less).Length of migration over 3 years was less likely to have a high nicotine dependence (3-5 years: OR=0.602, more than 5 years: OR=0.407 vs.<1 year) .Conclusion The rate of high nicotine dependence is high among rural-to-urban migrant workers in Shanghai.Level of education , duration of smoking and length of migration are significant influencing factors of nicotine dependence.
5.Survey on mental health status of rural-to-urban migrant workers in Shanghai
Tianhao WANG ; Zhigang PAN ; Yumiao WU ; Hua YANG ; Yao LIU ; Jian GONG ; Wei DAI ; Jing ZHOU ; Yao SHEN ; Shanzhu ZHU
Chinese Journal of General Practitioners 2014;(6):444-447
Objective To survey the mental health status of rural-to-urban migrant workers in Shanghai.Methods The survey subjects were selected from migrant workers in Shanghai with a multi-stage stratified cluster sampling method.The Symptom Check List ( SCL-90 ) scores were used for evaluation of metal health status.Results The prevalence of mental health problem of 5 626 rural-to-urban migrant workers in Shanghai was 18.8% ( 1 058/5 626 ).The mean total score of SCL-90 was 114.86 ±31.21.Compulsive-obsessive, interpersonal sensitivity, hostility ranked the top three among 9 factors, with scores of 1.37 ±0.43, 1.31 ±0.43 and 1.30 ±0.41 respectively.The mean total scores and scores of 9 factors of rural-to-urban migrant workers in Shanghai were lower than those of Chinese adults norm the mean total scores(129.96 ±38.76) ( t =-15.34, -3.84 --24.08 respectively, all P <0.01) .There were significant differences in mental health among migrant workers with different age , education , marital status , living conditions and occupation (all P<0.05).Conclusion The prevalence of mental health problems in Shanghai rural-to-urban migrant workers is high, with are related to their age, education, marital status, living conditions and occupation.
6.Effect of Heroin on DLG4 Expression in Hippocampus, Amygdala and Frontal Cortex of Rats.
Liang-ming LUO ; Qun GONG ; Jian-feng LIU ; Ming-quan ZHAO ; Dong-dong CHEN ; Yao-yao XIE ; Hua ZHU
Journal of Forensic Medicine 2015;31(3):185-199
OBJECTIVE:
To observe the expression of discs large homolog 4 (DLG4) protein in hippocampus, amygdala and frontal cortex of rats and evaluate postsynaptic density in heroin dependence.
METHODS:
The rat heroin dependent model was established by increasing intraperitoneal injection of heroin. DLG4 proteins in hippocampus, amygdala and frontal cortex of heroin dependent 9, 18, 36 days rats were detected with immunohistochemical staining and compared with that in the control group.
RESULTS:
DLG4 proteins in hippocampus, amygdala and frontal cortex were gradually reduced with extension of heroin dependent time.
CONCLUSION
Heroin dependence can affect postsynaptic density of hippocampus, amygdala and frontal cortex. The changes become more apparent with extension of heroin dependence time.
Amygdala/metabolism*
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Animals
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Disks Large Homolog 4 Protein
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Frontal Lobe/metabolism*
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Heroin/pharmacology*
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Heroin Dependence
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Hippocampus/metabolism*
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Injections, Intraperitoneal
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Intracellular Signaling Peptides and Proteins/metabolism*
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Membrane Proteins/metabolism*
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Rats
7.Safty evaluation, migration and distribution of human bone marrow derived-mesenchymal stem cells in the cns of young macaca fascicularis
Jiamei LI ; Hua ZHU ; Zhigang YAO ; Wei DENG ; Qin LI ; Chunmei MA ; Chuan QIN
Chinese Journal of Comparative Medicine 2016;26(9):7-12
Objective To evalutate the safty of hBMSCs transpalntation and to observe their migration and distribution in the brain of young macaca fascicularis. To establish a new technology platform and theoretical basis for the treatment of central nervous system diseases in children. Methods Labelled hBMSCs were transplanted into the striatum of young macaca fascicularis. Brain sections were examined to evalutate the inflammatory reaction and immunological rejection of local injection sites by HE observation and immunohistochemical staining. Migration and distribution of transplanted?hBMSCs was observed by real?time fluorescence quantitative PCR of male DNA and fluorescence microscope. Results The results showed that the direct intracerebral injection of hBMSCs did not cause systemic symptoms in animals. There is no inflammatory reaction and immunological rejection was detected, and degeneration and necrosis of neural cells and proliferation of glial cells were absent in the local injection sites. The transplanted hBMSCs survived, and migrated into the brain after 4 weeks transplantation. Its migration and distribution have certain regularity and were overlapping between transplant recipients. In addtion, hBMSCs tended to extend rostrally into the forebrain and showed preference of migrating toward the blood vessels and below the ependyma. Conculsions Intracerebral transplantation of hBMSCs is safe. And hBMSCs can survive and migrate into the brain.
8.Criteria for trainers of general practitioners in clinical training bases
Yao LIU ; Shanzhu ZHU ; Xiangjie ZHANG ; Hua YANG ; Jing ZHOU ; Qian CHEN ; Juan SHOU
Chinese Journal of General Practitioners 2014;13(7):534-538
Objective To develop criteria for trainers of general practitioners in clinical training bases.Methods A primary version of criteria was established through literature review and expert interview.Thirty-two experts with middle-level professional title or above,who had 10 years of working experience or more in general practice,teaching and administration,were invited for 3 rounds of Delphi consultation during March and October 2013.Results The criteria consisted of 3 first-grade indicators and 12 second-grade indicators were established.The first-grade indicators included professional quality,clinical competence and teaching capability.The weight coefficient of first-grade indicators were 0.332 0,0.336 0 and 0.332 0,respectively.For 3 rounds of consultation the activity coefficient of experts was all 100% ; the authority coefficients of experts were 0.881,0.897 and 0.883,respectively; and the harmonious coefficient of importance of the evaluation were 0.136,0.127 and 0.204,respectively (P < 0.01).Conclusion The established criteria are credible and important for the selection of trainers of general practitioners in clinical bases,which would improve the quality of standardized training for general practitioners.
9.Establishment of accreditation standards for trainers of general practitioners in community health service centers
Juan SHOU ; Yao LIU ; Xiangjie ZHANG ; Hua YANG ; Jing ZHOU ; Qian CHEN ; Shanzhu ZHU
Chinese Journal of General Practitioners 2014;13(8):634-638
Objective To establish a set of accreditation standards for trainers of general practitioner (GP) in community health service centers.Methods A modified Delphi expert consultation was conducted during December 2012 and September 2013.Thirty two experts of general practice from different teaching hospitals and community health service centers were invited for three rounds Delphi questionnaires.A set of accreditation standards for GP trainers in primary care was established through data analysis,synthesis and process.Results Expert activity coefficients were 100.0% for three rounds consultation,the authority coefficients were 0.894,0.882 and 0.893,respectively.The opinion coordination coefficients of experts were 0.156,0.166 and 0.215,respectively (P =0.000).The developed accreditation standards system comprised of three first-grade indicators and 14 second-grade indicators.The weight coefficients of three first-grade indicators,namely professional profiles,clinical competences in primary care and teaching abilities,were 0.339 8,0.335 6 and 0.324 6,respectively.Conclusions The established accreditation standard system is credible,which would appropriately guide the selection of GP trainers in primary care from three dimensions and enhance the quality of standardized training of general practitioners.
10.Relationship between genetic polymorphism of transcription factor 7-like 2 and genetic susceptibility to type 2 diabetes mellitus in Uygur population of Xinjiang region
Zhiqiang WANG ; Qi MA ; Tingting WANG ; Yinxia SU ; Yan MA ; Jun ZHU ; Hua YAO
Basic & Clinical Medicine 2015;(9):1162-1166
Objective To identify the potential association of transcription factor 7-like 2(TCF7L2 polymorphisms with type 2 diabetes mellitus in Uygur population of Xinjiang region .Methods In this case-control study ,819 ca-ses of type 2 diabetes mellitus patients were recruited in case group and 731 healthy individuals were selected as control.5 mL of blood sample were collected from each subject .The polymorphism was examined by matrix-assis-ted laser desorption/ionization-time of flight ( MALDI-TOF) and the OR value (95%CI) was evaluated by Logistic Regression Method to analyze the relationship between susceptibility to type 2 diabetes mellitus and different geno-types.Results In case group, the frequencies of TC, CC genotype and C allele at rs7901695 were higher than the corresponding frequencies in control group (P<0.05).The interaction between TCF7L2 and environment risk factors did not contribute to the occurrence of the type 2 diabetes mellitus .Conclusions The polymorphisms of rs7091695 in TCF7L2 but not rs7085532 in TCG7L2 may be associated with type 2 diabetes mellitus in Uygur pop-ulation in Xinjiang region .