1.Effect of bushen yizhi formula on neurotransmitter release in rat models with Alzheimer disease
Zhenguo ZHONG ; Maocai LIU ; Shilong LAI ; Jie GAO ; Shuyi CHENG
Chinese Journal of Tissue Engineering Research 2005;9(44):167-170
BACKGROUND: The decrease of activity level of choline acetyltransferase (ChAT) and reduction of membrane protein of synaptic vesicles are the main biochemical indexes in Alzheimer disease. Traditional Chinese bushen yizhi formula is of the effects of strengthening the kidney, replenishing qi, generating marrow and nourishing the brain, its medicine-containing serum can effectively protect these changes resulting from amyloid protein.OBJECTIVE: To investigate whether the medicine-containing serum of bushen yizhi formula can improve the pathological injury of neurons induced by amyloid protein metabolites in Alzheimer disease, thus to estimate the therapeutic effect of bushen yizhi formula on Alzheimer disease.DESIGN:A randomized and controlled trial. SETTING:Institute of Neuroscience, Second Affiliated Hospital, Guangxi College of Traditional Chinese Medicine, Nanning 530011, Guangxi Zhuang Nationality Autonomous Region, China; Encephalopathic Institute of Guangzhou University of Traditional Chinese Medicine.MATERIALS:The experiment was conducted from September 2003 to March 2004 at the Research and Development Center of New Drugs of Guangxi College of TCM. Totally 60 healthy Wistar rats of clean grade,three months, were at random divided as blank control and bushen yizhi groups, with 30 in each group. The bushen yizhi formula consisted of Fructus Lycii, Fructus Cnidii, Radix Gingeng, Radix Polygoni Multiflori,Cortex Moutan Radicis, Borneolum, etc. The medicine was prepared before use as 0.176 g/Ml liquid(including 1.13 g/Ml rude drugs).METHODS:① Each of the rats in bushen yizhi group was by garage given 1.0-1.5 Ml prepared liquid per day for consecutive 30 days. Within 4hours after the last administration the serum was isolated at 5 000 r/min for 5 minutes, filtered aseptically, the compliment was inactivated at 56 ℃,then the medicine-containing serum was obtained. The rats in blank control group were given saline of the same volume, and the serum was prepared with the same method. ② For NG108-15 cells, the concentration of amyloid protein 25-35 segments in the culture liquid was 5 μ mol/L, or 1-42 segments, 200 nmol/L, was suitable. ③ NG108-15 cells was first in cubated for a day in CO2 incubator with Dulbecco-improved Eagle medium of 0.1 molarity at 37 ℃, then 5 μmol/L amyloid protein 25-35 segment or 200 nmol/L segment 1-42 was added to continuously incubate for a day, after that the cell models of Alzheimer disease was ready. Then 50 g/L medicine-containing serum or Dulbecco-improved Eagle medium of blank control group serum was taken to replace the original culture liquid, and the amyloid protein segments of the above-mentioned concentration was still added for maintaining the cell models of Alzheimer disease, after 5 days'differentiation incubation, the indexes were detected with those in non-amyloid protein as control. ④ Western blotting, radioimmunoassay and electrophysiological examination were used to investigate the ChAT activity,synapsin protein level and rate of functional synapse formation of NG108-15cells in Alzheimer disease after treatment with the medicine-containing serum of bushen yizhi formula.MAIN OUTCOME MEASURES :The effects of the medicine-containing serum of bushen yizhi formula on ChAT activity, synapsin protein level,rate of functional synapse formation of NG108-15 cells and frequency of miniature plate potential in NG108-15 cells.RESULTS:Totally 60 rats involved all entered the final result analysis.① The effect of the medicine-containing serum of bushen yizhi formula on ChAT activity: In the conditions when there was no amyloid protein or the final concentration of amyloid protein 1- 42 was 200 nmol/L, the ChAT activity in bushen yizhi group was obviously higher than that in blank control group [(1651.2±134.5), (1336.1±268.2), ( 586.1±223.4),(1290.7±381.5)μmol/(g·h); P < 0.05] . ② The effect of the medicinecontaining serum of bushen yizhi formula on synapsin protein level: The absorbance values of synapsin Ⅰa and Ⅰb, in the conditions when there was no amyloid protein, amyloid protein 25-35 and 1-42, were respectively 5.02, 1.36 and 2.46 in bushen yizhi group; and 3.18, 0.57 and 0.71 in blank control group. The values in bushen yizhi group were respectively 1.6,2.4 and 3.5times those in blank control group. ③ The effect of the medicine-containing serum of bushen yizhi formula on functional synapse formation of NG108-15 cells: In the conditions when there was no amyloid protein or amyloid protein 1-42, the levels of functional synapse formation in bushen yizhi group wereall higher than those in blank control group [(90.6±6.0)%, (63.2±17.0)%, (58.0±13.1)%, (34.2±13.0)% ;P<0.05].④ The effect of the medicine-containing serum of bushen yizhi formula on frequency of miniature plate potential in NG108-15 cells: In the conditions when there was no amyloid protein or amyloid protein 1-42, the frequencies of miniature plate potential in bushen yizhi group were all higher than those in blank control group [(9.28±4.1), (5.48 ±5.14), (5.55±5.85),(3.05±4.46)/min; P < 0.01, P< 0.05].CONCLUSION:In the conditions when there existed no amyloid protein or amyloid protein, the medicine-containing serum of bushen yizhi formula can raise ChAT activity, not only correct the decrease of cellular synapsin caused by amyloid protein, but also raise the expression of synapsin protein, the rate of functional synapse formation and neurotransmitter release under the condition of no amyloid protein. It was shown that the prescription can antagonize the pathological development of Alzheimer disease, and play the therapeutic effect through enhancing the release power of neurotransmitter.
2.Modulation of microRNA-155 on CD4 + T lymphocyte in patients with unstable angina pectoris
Yongguang LU ; Lang LI ; Yanmei CHEN ; Jiming ZHONG ; Shuyi ZENG
Chinese Journal of Emergency Medicine 2011;20(9):960-965
ObjectiveTo inyestigate the effect of miRNA-155 expression on the differentiations and functions of CD4 + T lymphocyte in patients with unstable angina pectoris. MethodsTwenty-one patients with unstable angina pectoris (UAP) were enrolled in this study, and another 18 patients with normal coronary arteries evidenced by angiography were assigned as control group. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expression of miRNA-155 in CD4 + T lymphocyte of the peripheral blood. And the levels of IFN-γ and IL-4 in serum were determined by using enzyme-linked immunosorbent assay (ELISA). The CD4 + T lymphocyte were isolated from mononuclear cells prepared with Ficoll-Hypaque density-gradients centrifugation from human peripheral blood by magnetic cell sorting system. Then, the CD4 +T cells (2 × 106 cells/mL) were seeded in culture plates with 6 wells.The CD4 + T lymphocytes were divided into 3 groups in experiment: control group, miRNA-155 group, and miRNA-155 inhibitor group. The numbers of Th1 and Th2 cells were measured by flow cytometry analysis (FACS).The protein levels and expressions of IFN-γRα, T-bet, GATA-3 mRNA were measured by using western blotting and qRT-PCR, respectively. The levels of IFN-γ and IL-4 in culture supernatants of CD4 +T lymphocytes were detected by using ELISA. ResultsIn comparison with the control group, there was significant increase in the expression of miRNA-155 and serum level of IFN-γ in the UAP group (P <0. 01 ). There was a positive correlation between miRNA-155 and serum IFN-γ ( r =0. 842, P < 0. 0l ).However, in vitro, the number of Th1, the protein level and expression of T-bet mRNA, and supernatant IFN-γ increased, and the protein level and expression of IFN-γRα protein decreased in miRNA-155 group in comparison with the control group (all P <0. 01 ). Interestingly, there were no significant differences in the number of Th2 cells, the expressions of GATA-3mRNA and IFN-γRα mRNA, GATA-3 protein and supernatant IL-4 between control group and miRNA-155 group ( all P > 0. 05 ). And the miRNA-155 inhibitor could attenuate the effect of miRNA-155 effectively. ConclusionsThe miRNA-155 can promote differentiations and improve the function of Thl, playing an important role in the pathogenesis of unstable angina pectoris.
3.Effects of pre-existing antibody on seroconversion rate after influenza vaccination
Mao LI ; Yanhui CHENG ; Shuyi ZHONG ; Hejiang WEI ; Simin WEN ; Weijuan HUANG ; Dayan WANG ; Yuelong SHU
Chinese Journal of Microbiology and Immunology 2021;41(1):56-61
Objective:To investigate the effects of pre-existing antibody on seroconversion rate after influenza vaccination.Methods:This study recruited 1 900 healthy volunteers to receive influenza split vaccines in Xinjiang Uygur Autonomous region and Yunnan Province from September 2009 to October 2018. Hemagglutinin agglutination inhibition assay was used to detect the titers of specific antibodies in blood samples collected before vaccination and 28 d after vaccination and the effects of pre-existing antibody on the seroconversion to different influenza vaccine components were analyzed.Results:Trend analysis showed that with the increasing titer of pre-existing antibody, the seroconversion rates to A/H1N1, A/H3N2, B/Victoria and B/Yamagata vaccine components were gradually decreased (χ 2=121.76, P<0.001; χ 2=67.58, P<0.001; χ 2=45.25, P<0.001; χ 2=54.55, P<0.001). After adjusting for factors such as region, gender and age, multivariate logistic regression showed that pre-existing antibody titer equal to or higher than 40 was an independent factor that affected the seroconversion to A/H1N1, A/H3N2 and B/Victoria vaccine components, and the adjusted OR (95%CI) values were 2.50(2.00-3.13)、1.64(1.35-2.00) and 2.50(1.79-3.45), respectively. Conclusions:The seroconversion rate to each vaccine component was negatively correlated with the pre-existing antibody titer. The factor that pre-existing antibody titer equal to or higher than 40 was detrimental to the seroconversion to A/H1N1, A/H3N2 and B/Victoria vaccine components, but had no significant influence on B/Yamagata seroconversion.
4.Comparison of the Effect of Different Establishment Methods on the Rat Models of Polycystic Ovary Syndrome
Zhengwang YANG ; Fang ZHOU ; Zhenyu TAN ; Qiongfang XU ; Shuyi ZHANG ; Fei ZHONG ; Lan LI ; Juan XIONG ; Ximing YU ; Yaobang LU
Acta Laboratorium Animalis Scientia Sinica 2010;18(1):13-16,彩4
Objective To establish rat models of polycystic ovary syndrome(PCOS) induced by different methods,to assess the serum levels of several related hormones,to examine the morphological changes in the ovaries,and to discuss their significance.Methods Letrozol,sodium prasterone sulfate,or sodium prasterone sulfate combined with human chorionic gonadotropin were used to establish rat models of PCOS.Radioimmunoassay was used to measure the serum levels of hteinizing hormone(LH),follicle-stimulating hormone(FSH),estrogen(E_2),progesterone(P),testosterone(T),prolactin(PRL),and insulin(INS).HE staining was used to examine the morphological changes of the ovaries.Results Comparing with the normal group A,the serum FSH was increased and the serum progesterone was reduced in the group B,with a statistically significant difference(P<0.05).The serum testosterone was significantly higher in the group B than in the group A(P<0.01).The levels of serum sex hormones and insulin were not significantly different in the group D and C(P>0.05).In comparison with the group C,the levels of serum testosterone and LH/FSH ratio was significantly increased in the group E.(P<0.05).Comparing with the group D,the serum levels of progesterone and testosterone were significantly increased in the group E(P<0.05).The ovaries in the rats of groups A and C showed almost a normal histyology,with mature follicles and dominant follicles.Polycystic changes were observed only in the ovaries of groups B,D and E.Conclusion At the aspect of affecting the level of sex hormones in serum and changing the ovarian morphology.adopting letrozol tablets or sodium prasterone sulfate combined with HCG to induce rat PCO model is more close to clinic manifestations and meets the criteria of PCO animals.In the rat PCOS models induced with letrozol or with sodium prasterone sulfate combined with HCG,either the serum levels of sex hormones and ovarian histology are quite similar to those of human clinical appearance,and may well meet the modeling requirements for future experimental studies of polycystic ovary syndrome.
5.Progress in the role of leptin in immune response
Mao LI ; Simin WEN ; Shuyi ZHONG ; Huanle LUO ; Yuelong SHU
Chinese Journal of Microbiology and Immunology 2020;40(5):387-392
Leptin, a peptide hormone discovered in the 1990s, arouses interest as it can regulate the body′s metabolism. In recent years, many studies have shown that leptin can promote the proliferation, activation and cytokine synthesis of various immune cells, and participate in innate and adaptive immune responses. This article reviewed the role of leptin and involved signaling pathways in immune response and the potential of leptin as a vaccine adjuvant.