1.Identification of human leucocyte antigen -A* 0201-restricted hepatitis C virus-cytotoxic T lymphocyte epitopes
Jun CHEN ; Zhiliang DUAN ; Wenci GONG ; Yongping CHEN ; Lifang ZHANG ; Qin ZHANG ; Jinsheng WEN
Chinese Journal of Infectious Diseases 2010;28(11):651-655
Objective To identify human leucocyte antigen (HLA)-A* 0201-restricted hepatitis C virus (HCV)-cytotoxic T lymphocyte (CTL) epitopes. Methods Based on the prediction results of RANKpep and SYFPEITHI prediction programs, six candidate CTL epitopes were selected and synthesized. The affinity of candidate CTL epitopes to HLA-A* 0201 molecules of T2 cells was explored. Subsequently, enzyme-linked immunosorbent spot (ELISPOT) assay and intracellular cytokine staining (ICS) were utilized to determine whether candidate CTL epitopes could induce the recall positive response in peripheral blood mononuclear cells (PBMC) of HLA-A* 0201 positive HCV-1b-infected patients. Results Among six candidate CTL epitopes, peptides C_181(LLSCLTTPV) and NS2_172 (VLQAGLIRV) had high affinity to HLA-A* 0201 molecules. Moreover, the affinity was proportional to the concentration of peptide. Furthermore, among ten HLA-A* 0201 positive HCV-1b-infected patients, the frequencies of C_181 and NS2_172-specific interferon (IFN)-γ-producing cells were 0-19 spots forming cells (SFC)/1 × 105 PBMC and 0-20 SFC/1 × 105 PBMC, respectively.The percentages of C_ 181 and NS2_172-specific IFN-γ+ CD8+ T lymphocytes in total CD8+ T lymphocytes were 0.006%-0.065% and 0.005%-0.080%, respectively. Conclusion Peptides C_181 (LLSCLTTPV) and NS2_172 (VLQAGLIRV) are identified as novel HLA-A* 0201-restricted HCV-CTL epitopes.
2.Hypolipemic treatment of hyperlipidemic pancreatitis with enema
Yunxing SHI ; Jiao Lü ; Guozhong ZHOU ; Yongping LI ; Changyun LIU ; Linhua QIN ; Zhiwu ZHENG
International Journal of Traditional Chinese Medicine 2010;32(3):247-248
Objective To investigate hypolipemic treatment of hyperlipidemic panereatiti(HLP)with integrated traditional Chinese and western medicine.Methods Cinical data of 20 patients of HLP were analyzed retrospectively.Eight patients in the control group were treated with conventional therapies,while 12 patients in the treatment group were treated as follows:①Enema with 180ml solution(50% magnesium sulfate 30 ml.Glycerin 60ml,water 90ml).②Rhubarb gastrogavage with 9 g tid.③Intravenous drip with 24 g salvia miltiorrhiza qd.Results The treatment group had significant difference comparing with the control group in terms of the serum TG in 48 hours(P<0.01),time of autonomous bowel movement recover(P<0.01),days of abdominal pain disappear(P<0.05),days of hospitalization(P<0.01).Conclusion The treatment of Enema with 180 ml solution.Rhubarb gastrogavage with 9g tid,and Intravenous drip with 24 g salvia miltiorrhiza qd can relieve the symptoms of HLP and decrease blood-fat greatly.
3.Expression and significance of CXCL5 and CXCL8 in hepatic fibrosis and cirrhosis tissues
Xinxin LIAO ; Caixian LIAO ; Yongping HUANG ; Ancheng QIN ; Jie YUAN ; Yongqiang LAI ; Zuyuan GONG
Chinese Journal of Digestive Surgery 2010;9(2):130-132
Objective To investigate the level of chemotactic factors(CXCL5 and CXCL8)in hepatic fibrosis and cirrhosis tissue.Methods Hepatic tissues were obtained from 9 patients with hepatic hemangioma (hepatic hemangioma group),10 patients with liver fibrosis(liver fibrosis group)and 11 patients with liver cirrhosis(1iver cirrhosis group)at Nanfang Hospital from May 2008 to May 2009.The contents of CXCL5 and CXCL8 in hepatic tissue were assayed by ELISA.All data were analyzed by one-way ANOVA,Pearson rank correlation or Spearman correlation.Results The contents of CXCL5 and CXCL8 were(0.8±0.7)ng/g and(6.2±3.7)ng/g in hepatic hemangioma group,(2.0±2.0)ng/g and(11.6±3.5)ng/g in liver fibrosis group and (17.1±4.8)ng/g and(12.3±3.9)ng/g in liver cirrhosis group,with significant difference among the 3 groups (F=60.050,7.690,P<0.05).The expression of CXCL5 was correlated with the content of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and prothrombin time(PT)(r=0.502,0.468,0.523,P<0.05):the expression of CXCL8 was correlated with the content of ALT,AST.total bilirubin and PT(r=0.477,0.504,0.537,0.431,P<0.05).Conclusions With the aggravation of hepatic fibrosis,the contents of CXCL5 and CXCL8 are increased with different patterns.The changes of CXCL5 and CXCL8 are related with the injury of liver,but the changes of CXCL5 and CXCL8 do not correspond with the degree of the injury of liver.
4.Evaluation of the effect of post-abortion counseling and education among unmarried abortion adolescents
Junpu QIN ; Shan CHEN ; Na DI ; Yongping YANG ; Li ZHOU ; Dikai ZHANG
Chinese Journal of Obstetrics and Gynecology 2010;45(3):201-204
Objective To find out the requirement and to evaluate the effect of post-abortion counseling and education (PACE) among unmarried abortion adolescents.Methods The subjects of the study were unmarried adolescents from 10 to 24 years of age who wanted induced abortion in the Second Affiliated Hospital of Sun Yat-sen University from December 2007 to April 2008.Totally 122 subjects received the intervention of PACE were considered as intervention group.Meanwhile,67 subjects refused the intervention of PACE were considered as no intervention group.Two groups were both investigated the requirements of PACE before abortion and were followed-up at one year after abortion.Results 97.4% (184/189) of 189 unmarried abortion adolescents were willing to receive PACE,48.1% (91/189) of them hoped to receive PACE when saw the doctor,72.0% (136/189) of them required face-to-face counseling during PACE.During the year after abortion,74% (57/77) cases in intervention group and 24% (10/41) cases in no intervention group took effective contraception (P< 0.01 ),while 1% (1/77) of intervention group and 10% (4/41) of no intervention group had unwanted pregnancy.There were significant different for the rate of unwanted pregnancy between two groups (P=0.034).Conclusion For unmarried abortion adolescents,the intervention of PACE may markedly increase the rate of effective contraception used and decrease the rate of another unwanted pregnancy.
5.Germination and propagartors of artificial seeds of Dendrobium huoshanense.
Ziqing QIN ; Ting ZHAO ; Jing QIU ; Yi LIN ; Yongping CAI
Chinese Journal of Biotechnology 2008;24(5):803-809
The artificial seeds of Dendrobium huoshanese was produced with Auxiliary buds, Protocorm-like bodies, and adventitious shoots. By using orthogonal experiment, we studied the effect of Maltose (%), hormone rate between 6-BA (mgx L(-1)) and NAA (mgL(-1)), active carbon (%), sodium alginate (%), time of ion exchange (min) on germination rate of artificial seeds of D. huoshanese. Then the leaking rate of maltose and variation of pH value of artificial seed capsule during vegetating of artificial seeds of D. huoshanese was measured. The results show that maltose played the most important role in inducing D. huoshanese artificial seeds to germinate. The optimal combination was: maltose 4%, hormone rate between 6-BA (mg x L(-1)) and NAA (mg x L(-1)) 10:1, active carbon 0.1%, sodium Alginate 4%, time of ion exchange is 10 min. Protocorm-like bodies were appropriate propagartor, the germination rate of artificial seeds of D. huoshanese takeing Protocorm-like bodies as the propagartors is 90.1%. After germination, the survival rate of seedlings of artificial seeds was 80.6%, the leaking rate of maltose of artificial seed capsule was 0.52%, and the pH value of artificial seed capsule decreased during the process of vegetation of artificial seeds. After having been stored at 4 degrees C for 20 d, the germination rate of artificial seeds of D. huoshanese takeing Auxiliary buds, Protocorm-like bodies, Adventitious shoots as the propagartors were 3.3%, 10.6%, 5.2%. Under natural conditions the germination rate was 13.8% after 10.0 g/L carbendazim was appended into artificial seed capsule. This result provides a foundation of manufacture and further study of the artificial seeds of D. huoshanese.
Alginates
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pharmacology
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Charcoal
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pharmacology
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Culture Techniques
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methods
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Dendrobium
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growth & development
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Germination
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Glucuronic Acid
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pharmacology
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Hexuronic Acids
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pharmacology
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Maltose
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pharmacology
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Plant Shoots
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growth & development
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Seeds
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growth & development
6.Andrographolide inhibits extracellular signal-regulated kinase 1/2 signaling pathway in activated macrophages.
Linhua QIN ; Jiao Lü ; Lin KONG ; Yunxing SHI ; Yongping LI ; Guozhong ZHOU ; Zhiwu ZHENG ; Lin LI ; Xingying JI
Journal of Integrative Medicine 2011;9(6):632-637
Objective: To investigate the effects of andrographolide on extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway and tumor necrosis factor-α (TNF-α) expression in lipopolysaccharide (LPS)-activated macrophages. Methods: LPS-activated mouse peritoneal macrophages were cultured in media with different concentrations of andrographolide. Cytotoxicity of andrographolide was detected by cell counting kit-8. The macrophages were lysed, and then expressions of phosphorylated ERK1/2, JNK and p38 and nuclear factor-κB inhibitor (IκBα) protein were detected by Western blotting and TNF-α mRNA expression was detected by reverse transcription-polymerase chain reaction. Supernatants of the macrophages were used to detect content of TNF-α protein by enzyme-linked immunosorbent assay. Results: Andrographolide at 1-100 μg/mL showed no cytotoxicity on LPS-activated mouse peritoneal macrophages. Andrographolide inhibited ERK1/2 phosphorylation in LPS-activated murine peritoneal macrophages, which was concentration-dependent (P<0.01). Andrographolide at 1-25 μg/mL had no effects on phosphorylation levels of JNK and p38 and IκBα degradation in LPS-stimulated mouse peritoneal macrophages. In activated macrophages, TNF-α expression was inhibited by 12 μg/mL andrographolide and 20 μmol/L PD98059 (inhibitor of ERK1/2 signaling pathway) at both mRNA expression and protein secretion levels. Conclusion: In LPS-activated macrophages, andrographolide may inhibit the expression of TNF-α by inhibiting ERK1/2 signaling pathway.
7.Pharmacokinetics and relative bioavailability study of roxithromycin tablet in Chinese healthy volunteers by LC-MS/MS.
Li ZHENG ; Yongping QIN ; Feng NAN ; Ying WANG ; Nan XU ; Maozhi LIANG
Journal of Biomedical Engineering 2009;26(6):1315-1319
This is a study of assessing the comparative bioavailability of roxithromycin produced by two companies in 36 healthy volunteers. On the basis of informed consent, 36 healthy male volunteers received each medicine at the roxithromycin dose of 150mg in a cross-over study. There was a 1-week washout period among the doses. Plasma concentrations of roxithromycin were monitored by an LC-MS/MS for over a period of 72 hours after administration. In this study, roxithromycin was generally well tolerated. After an oral administration of roxithromycin capsule, the pharmacokinetic parameters of roxithromycin, such as AUC(0-72 h) (66 076 microg x L x h(-1) and 70 334 microg x L x h(-1) for test and reference capsule, respectively) and AUC(0-infinity) (68 153 microg x L x h(-1) and 72 362 microg x L x h(-1)) were significantly similar. For test and reference capsule, the values of C(max) were 6 631.5 microg x L(-1) and 7 033.9 microg x L(-1) respectively, of T1/2 were 15.39 +/- 4.61 h and 16.06 +/- 5.56 h, and of T(max) were 1.3 +/- 0.9 h and 1.4 +/- 0.7 h respectively. The relative bioavailability F was 94.9% +/- 22.4% of tested formulation. The values of 90% confidence interval around the ratios (test/reference) (obtained by analysis of variance, ANOVA) were 88.3%-101.2% for C(max), 86.2%-98.9% for AUC(0-72) h, being within the predefined acceptable range for the conclusion of bioequivalence. The results of statistical analysis suggest that the two formulations be bioequivalent.
Adult
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Anti-Bacterial Agents
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administration & dosage
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pharmacokinetics
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Biological Availability
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Chromatography, High Pressure Liquid
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methods
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Cross-Over Studies
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Humans
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Male
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Roxithromycin
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administration & dosage
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pharmacokinetics
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Tablets
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Tandem Mass Spectrometry
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methods
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Young Adult
8.Establishing an organic model of SMC proliferation with cultured aorta of rats and exploring the underlying mechanism.
Yanlin ZHANG ; Caiying WANG ; Qin YANG ; Yaan YANG ; Yongping GU ; Mincheng WANG ; Kaiyun WU
Journal of Biomedical Engineering 2008;25(6):1405-1410
To study the mechanism of proliferous vascular disease as well as its prevention and treatment, an organic model was established with cultured aortas of rats, and the mechanism there-in invloved was probed. Immunostaining histology showed that smooth muscle cell (SMC) proliferation was observed in the aorta segments of rats, after their endothelia being injured and cultured in vitro with 20% fetal bovine serum. After being cultured for 5 days, various degrees of proliferation of SMC on cultured artery segments were observed by HE staining, and conspicuous plaques were developed after being cultured for 13 days. The proliferous SMC was also observed by Brdu labeling. RT-PCR examination showed that the mRNA expression of hypertension-related gene-1 (Hrg-1) and smooth muscle 22 alpha (SM22a) in the aortas decreased with the prolongation of culture time, and completely disappeared after being cultured for 13 days . But when cultured in vitro for ten days, the ET-1 content of supernatant and the proliferous SMC labeled by Brdu increased obviously and the expressions of Hrg-1 and SM22a decreased after the endothelium was destroyed. Compared with the injured endothelium groups, the proliferous SMC of injured endothelium plus BQ123 groups decreased visibly. The same significant differences between serum groups and serum-free groups were also observed. These results suggest that the culturing of rat aorta segments in vitro can induce the proliferation of SMC and the transform of phenotype from contractile type to synthetic type. The ET-1 and serum are the main factors in the proliferation of SMC and in the transform of phenotype. This organic model could serve as a good experimental platform for the researches into the mechanism of proliferous vascular disease as well as its prevention and treatment.
Animals
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Aorta, Abdominal
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cytology
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Cell Proliferation
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Disease Models, Animal
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Endothelin-1
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genetics
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metabolism
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Female
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Male
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Microfilament Proteins
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genetics
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metabolism
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Muscle Proteins
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genetics
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metabolism
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Muscle, Smooth, Vascular
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cytology
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Organ Culture Techniques
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RNA, Messenger
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genetics
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metabolism
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Rats
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Rats, Sprague-Dawley