3.Preliminary analyses of lymphocyte differentially expressed proteins in Huaihua Dong peoples with hypertension
Ling JIN ; Weiwu LIU ; Zi XING ; Yufeng DUAN ; Hongliang LI
Journal of Chinese Physician 2013;(5):599-603
Objective To explore lymphocyte differentially expressed protein profile and its pathogenic significance in Huaihua Dong peoples with hypertension.Methods The lymphocyte protein profiles among 130 cases of blood specimen were detected and identified with matrix-assisted laser desorption ionization/time of flight mass spectrometry (MALDI-TOF-MS) and Mascot search against protein database.Results The reproducible protein-maps of two-dimensional gel electrophoresis were obtained among three groups.Compared with Dong normal peoples,the protein disulfide isomerase-related protein 5 and heat shock protein 27 showed higher expressions in the Dong peoples with hypertension (P < 0.01),but chain A,structure of haemoglobin in the deoxy quaternary state with ligand bound at the alpha haems is no statistical significance(P >0.05).Those three proteins were upregulated in Han peoples with hypertension (P <0.01).Conclusions The up-regulation of the protein disulfide isomerase-related protein 5 and heat shock protein 27 in the Dong and Han peoples with hypertension suggest that those two proteins might be associated with the occurrence and development of hypertension.
4.A Meta -analysis of Radix Astragali f or Primary Nephrotic Syndrome in Adu lts
Junming FAN ; Liansheng LIU ; Zi LI ; Baihai SU ; Jin GUAN
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(01):-
Objective To evaluate the effects of Radix Astr agali combined with prednisone and i mmunosuppressant for primary nephrotic syndrome (PNS)in adults and to compare the effects o f Radix Astragali in various prepata tions for PNS.Methods Randomized controlled trials were a pplied for systemic reviews.Electr onic and manual retrieve of Medline,Embase,Cochrane Library,CBMdisc a nd CEBM/CCD and relevant medical jou rnals in China were applied to search the RCTs of Radix Astragali,non -specific treatment,glucocorticoids and i mmunosuppresants for PNS,and the RCTs were analysed with RevMan 4.1.Results There were 14randomized controlled trials with 524cases involved.Meta-analysis showed that Radix Astragali could in crease the therapeutic effect of pre dnisone and immunosuppressant for PNS and re-duce its recurrence.Radix Astragali also had an effect in decreasing 24-hour proteinuria content and the pla sma levels of total cholesterol and albumin.There were no differences between single injection and compound decoction.Asymmetry showed in"Funnel plot"may be related to publication bias,l ow quality of methodology and small -size in sample.Conclusion Radix Astragali and its prescriptio n may become a prospect therapy for PN S and its recurrence and the com-bination of traditional Chinese med icine and western medicine can be more effective for PNS.The dedinite effect of Radix Astragali for PNS will be further con firmed by multiple -center,large -s ample randomized controlled trial.
5.Serum-Free Culture and Identification of Neural Stem Cells from Hippocampus of Neonatal Rats
zi-jin, YANG ; hai-yan, WANG ; li-min, WANG
Journal of Applied Clinical Pediatrics 2006;0(14):-
Objective To explore the best serum-free cultural way in neural stem cells from the hippocampus of the newborn rats and observe the characteristics of growth,proliferation,and induced differentiation of neural stem cells.Methods The neonatal rats′ hippocampus tissues were dissociated mechanically.The cells were cultured in the serum-free cultural medium which were added separately basic fibroblast growth factor(bFGF),epithelium growth factor(EGF),bFGF+EGF by suspended cultural way.The neural stem cells were identified by nestin immunofluorescence.After induced differentiation of embryonic cow serum,the differentiated cells were identified.Results The serum-free medium added bFGF and EGF could perfectly induce neural stem cells to proliferate and the differentiated cells expressed the specific antigen of neurons,astrocytes and oligodendrocytes.Conclusion The serum-free medium with bFGF and EGF can be used to culture neural stem cells from hippocampus tissue of rats in vitro.
6.Value of Application of Histamine Provocation Test and Airway Resistance Detection in Diagnosis and Therapeutic Efficacy in Preschool Children with Asthma
xi-zhe, YUAN ; hong-zi, LI ; zheng, JIN ; ling, NIE
Journal of Applied Clinical Pediatrics 2006;0(16):-
Objective To study the value of application of histamine provocation test and airway resistance measurement in diagnosis and therapeutic efficacy in preschool children with asthma.Methods Histamine provocation test and airway resistance measurement by the Italian MEFAR MB3 provocating instrument and Germen Microloop lung function instrument for 42 cases who were diagnosed as asthmatic(27 patients with bronchial asthma and 15 cases of cough variant asthma)and 21 healthy cases was compared,and the differences between the 2 groups and the value of therapeutic efficacy were analyzed.Results The resistance ratio of respiratory tract of control group was(97.11?9.09)%,which in asthma and cough variant asthma group was(229.37?57.48)% and(248.80?76.80)%.There was significant difference between the 3 groups(F=48.466 P
7.Nerve Stem Cells Orientation Differentiation in Neonatal Rat Hippocampus Induced by Brain-Derived Neurotrophic Factor in Vitro
jun-feng, YU ; zi-jin, YANG ; wei-hong, ZHANG
Journal of Applied Clinical Pediatrics 2006;0(14):-
Objective To explore the influence of brain-derived neurotrophic factor (BDNF) on the differentiation of nerve stem cells (NSCs) from neonatal rat hippocampus in vitro and to find new revulsant of NSCs,which can improve the percentage of NSCs differentiating into neurons.Methods Twenty-four hours neonatal rats were selected to obtain hippocampus tissue to culture NSCs in serum-free culture medium by suspending culture.The high pure NSCs were obtained after passing 2 generations.The culture cells were identified as NSCs by staining of nestin,which was NSCs special marker.After passaged three generations,the NSCs were randomly classified into 2 groups:test group and control group.There were 15 pieces per group.There was 2 mL per piece,which contains 1?105 cells.50 g/L fetal bovine serum(FBS) and 20 ?g/L BDNF were added into foundational culture medium in test group;only 50 g/L FBS was added into foundational culture medium in control group.The neurons and their percentage were tested using the immunofluorescence labeling and flow cytometer after 7 days of differentiated cultivation.Results The hippocampus tissue cells grew in globular in serum-free culture medium by suspending culture,which expressed highly positive by nestin immunofluorescence staining.Its purity was above 90%.The percentage of neurone specific enolase(NSE)-positive cells in test group was 60.45%,which was obviously higher than that of control group (23.67%).The difference was significant between 2 groups(?2=27.75 P
8.Association of Hepatitis B virus infection and the expression of Toll-like receptors 2 and 4 in HepG2 cells
Sheng JIN ; Da-Zhi ZHANG ; Ya-Zi CHEN ;
Chinese Journal of Infectious Diseases 1997;0(04):-
Objective In order to explore the roles of TLR2 and TLR4 in the hepatocyte dam- age caused hy hepatitis B virus infection,and to find whether LPS can affect the damage of hepato- cytes pre-and pos-HBV infection,we detected the changes of TLR2 and TLR4 expressions in hu- man hepatocyte lines HepG2 cells and 2.2.15 cells.Methods HepG2 ceils are most similar to normal human hepatocytes and 2.2.15 ceils are HepG2 cells infected with HBV.We selected these two cell lines to study the differences of TLR2 and TLR4 expression between HepG2 cells before and after HBV infection.In this research,both HepG2 and 2.2.15 cells were stimulated with 0?g/ml, 1?g/ml,10?g/ml,100?g/ml,1 mg/ml and 10 mg/ml LPS.Then the expression of protein of TLR2 and TLR4 were examined by immuno-histochemistry(IHC).The cnRNA of HepG2 and 2.2.15 ceils stimulated with 0?g/ml and 10 mg/ml LPS were examined by reversal transcription-pol- ymerase chain reaction(RT-PCR).Thereafter,the apoptosis of HepG2 and HepG2.2.15 cells were examined by flow cytometry(FC),and the expressions of HBsAg and HBeAg of HepG2.2.15 cells tested with Abbott kits.Results IHC and RT-PCR analysis revealed that TLR2 and TLR4 expres- sions could he detected in both HepG2 and 2.2.15 cells.Moreover,without immune activation, TLR2 and TLR4 expressions were higher in the presence of higher concentrations of LPS.FC analy sis revealed that no apoptosis detected in HepG2 ceils stimulated with LPS in this research,but apop- tosis could be detected in 2.2.15 cells when treated with the same factors.Furthermore,the apoptosis ratios increased with the increase of LPS concentrations.When concentrations of LPS were 1?g/ml, 10?g/ml,100?g/ml,1 mg/ml and 10 mg/ml,the apoptosis ratios were 1.94%,3.03%,3.50%, 3.72%,5.30%,respectively.Abbott analysis revealed that expressions of HBsAg and HBeAg of 2.2.15 cells stimulated with LPS were lower than those not stimulated with LPS.Conclusion HBV can affect the expressions of TLR2 and TLR4 in HepG2 cell lines.LPS can lead 2.2.15 cells to apop- tosis but not HepG2 cells.Although LPS cannot damage normal hepatocytes,it might aggravate hep- atocytes damage when their microenvironment was changed by HBV infection.
9.Construction of HIV-2 gag recombinant fowlpox virus and its immune reactivity in mice
Li-Shu ZHANG ; Ning-Yi JIN ; Zi-Jian LI ;
Chinese Journal of Infectious Diseases 1997;0(04):-
Objective To provide experimental evidences for developing a safe and effective re- combinant fowlpox virus which can prevent the infection of HIV-2.Methods A fowlpox virus(FPV) transferring vector was constructed by inserting HIV-2 gag gene to the downstream of a synthetic complex promoter ATI-p7.5?20 of vector pUTA2.Transfection was then carried out,and recombi- nant FPV(rFPV)was screened by 5'-bromo-deoxyuridine(BrdU),genome PCR and western blot detection.Balb/c mice were immunized with rFPV by muscular injection.Anti-HIV-2 antibody, CD4~+ and CD8~+ T-cell count and specific target-killing activity of spleen CTL in immunized mice were analyzed by ELISA,FACS and LDH release assay,respectively.Results A transferring vector pA- gag was constructed and confirmed by amplifying a fragment of 766 bp from the rFPV genome.Mean- while,HIV-2 multi-antibody-specific protein blot(55 000)was detected from the recombinant virus and the HIV-2 specific antibody was detected from the immunized Balb/c mice.HIV-2 specific target- killing activity of spleen CTL was observed in immunized mice.Conclusion A recombinant fowlpox virus expressing HIV-2 structural protein Gag has been obtained,and it can stimulate HIV-2-specific eelluar and humoral immune reactions in mice.