5.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.
6.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.
7.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.
8.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
9.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
10.Influence of glycine supplement on partial biochemical indicator in exhausive exercise mice
Jin CAO ; Xiaofei FAN ; Liangju LI ; Zi XU
Chongqing Medicine 2015;(31):4327-4329,4332
Objective To investigate the influence of glycine supplement on partial biochemical indicator in exhaustive exer‐cise mice .Methods Divided Kunming′s male‐mice into 3 group(10/group):exhaustive swimming group(E) ,exhaustive swimming group with glycine supplement(G) and silent control group (S) .Mice in the G group was treated with 4‐weeked intragastric admin‐istration of glycine ,while mice in other 2 groups received physiological saline .After 4 weeks ,mice in E and G group did the exhaus‐tive swimming exercise .After sampling ,the activity of complement hemolytic activity was measured ,and the serum levels of some biochemical indexes were determined .Results Time of exhaustive swimming in G group was significantly longer than E group(P<0 .01) .After exhaustive swimming ,the ALT ,AST ,ALP ,LDH ,CK ,N ,Fe3+ and CHA in E group were significantly changed com‐pared with S group(P<0 .01);while those indicators in G group were significantly improved than E group (P<0 .01) ,the ALP and LDH level can regain to the level of S group (P>0 .05) .Conclusion Exhaustive swimming mice can have part indicator abnormali‐ty ,but Glycine supplement can effectively improve that condition and enhance the exercise ability to some extent .