AIM: To study the apoptotic effect of TNFα on rat pulmonary microvascular endothelial cells (PMVEC) and the influences of Fas, NF-κB in its mechanism. METHODS: Apoptosis of PMVEC was analyzed and quantitated with TUNEL, flow cytometer. The distribution of NF-κB was detected via histoimmunochemical staining in TNF-treated cells and the control. Northern blot was applied to assess the influence of TNF on PMVEC Fas expression. Fas antibody was used to investigate the apoptotic effect of Fas on PMVEC. Activation of caspase-8 was detected with Western blot. Expression of caspase-3 was analyzed with histoimmunochemical staining. RESULTS: After treatment with 5×108 U/L TNF for 24 hours, viable PMVEC significantly diminished. Apoptosis rate was 14.0%±3.1% detected with TUNEL, and 13.1% with flow cytometer. Histoimmunochemical staining showed that NF-κB relocated from cytoplasm to the nuclear. When the cells were co-cultured with TNF and APDC, an NF-κB inhibitor, less cells were viable and more cells were positively stained with TUNEL. Fas expression in PMVEC was elevated treated with TNF. Apoptosis in PMVEC was found aggravated, when the cells were co-cultured with TNF and anti-Fas antibody. The positive rate was 24.1%±1.5% with TUNEL. Increase of caspase-8 activation was manifested by Western blot following TNF stimulation. Caspase-3 expression was found elevated using histoimmunochemical staining. Cell permeable caspase-3 inhibitor significantly ameliorated PMVEC apoptosis induced by TNF. CONCLUSION: 1. Large dose of TNF(5×108 U/L) can induce apoptosis in rat PMVEC. 2. NF-κB has a protective effect on PMVEC apoptosis. 3. TNF up-regulates Fas expression in PMVEC. And the latter takes a part in apoptosis. 4. TNF induced caspase-8 activation in PMVEC, and more caspase-3 was expressed. These may be involved in PMVEC apoptosis induced by TNF.

AIM:To study the apoptotic effect of TNF? on rat pulmonary microvascular endothelial cells (PMVEC)and the role of Fas, NF-?B in its mechanism. METHODS: Apoptosis of PMVEC was analyzed and quantitated with TUNEL, flow cytometer. Northern blot was applied to assess the influence of TNF? on PMVEC Fas expression. Fas antibody was used to investigate the apoptotic effect of Fas on PMVEC. Activation of caspase-8 was examined by Western blot. Expression of caspase-3 was analyzed with histo-immunochemical staining. RESULTS:Growth curve showed that TNF? suppressed endothelial cell proliferation in a dose-dependent manner. After treatment with 5?10 6 U/L TNF?, apoptotic rate was 14.0%?3.1% detected with TUNEL, and 13.1% with flow cytometer. When the cells were co-cultured with TNF? and APDC, an NF-?B inhibitor, less cells were viable and more cells were positively stained with TUNEL. Fas expression in PMVEC was elevated after TNF? treatment. Co-culturing with Anti-Fas antibody aggravated PMVEC apoptosis. Caspase-8 activity and caspase-3 expression was elevated. Caspase-3 inhibitor significantly ameliorated PMVEC apoptosis. CONCLUSION: Large dose of TNF? (5?10 6 U/L) can induce apoptosis in rat PMVEC. NF-?B has an anti-apoptotic effect in PMVEC. TNF? up-regulates Fas expression in PMVEC, and the latter takes a part in apoptosis. Caspase-8 and caspase-3 are involved in PMVEC apoptosis induced by TNF?.

OBJECTIVE:A RP-HPLC method was established to determine the concentration of ambroxol hydrochloride in human plasma.METHODS:Waters HPLC instrument was used with the Hypersil BDS C 18 column(5?m,250mm?4.6mm). Ditiazem hydrochloride was chosen as the internal standard.The mobile phase was composed of acetonitrile-methyl alcohol-0.01mol/L phosphatic buffer(pH7.0)-tetrahydrofuran(35∶35∶27.5∶2.5).Flow rate was1.0ml/min.Detection wavelength was242nm.A single oral dose of90mg imported ambroxol hydrochloride tablet was given to9healthy male volunteers.Ambro xol concentration in plasma was assayed.RESULTS:The standard curve of ambroxol hydrochloride was linear in the range of10～640ng/ml.The minium detection concentration was10ng/ml.The extraction recovery was more than90%.A one-compartment open pharmacokinetic model was adopted in ambroxol plasma concentration-time data analysis.The main pharmac okinetic parameters were as follows:C max =(226.53?34.59)ng/ml,T max =(1.82?0.80)h,T 1/2ke =(6.27?0.66)h,AUC 0～24 =(2363.55?448.86)ng/(h?ml).CONCLUSION:The method is simple.The sensitivity and accuracy are high.It is applicable to study the pharmacokinetics of ambroxol hydrochloride in healthy male volunteers.

Aim To investigate the pharmacokinetics of intravenous injection of magnesium isoglycyrrhizinate in single dose in Chinese healthy volunteers.Methods 9 healthy volunteers received magnesium isoglycyrrhizinate injections in single dose of 100,200,300 mg respectively.The concentrations of magnesium isoglycyrrhizinate in plasma at different time were assayed with HPLC-UV method.The pharmacokinetic parameters of magnesium isoglycyrrhizinate injections were calculated with program 3P87.Results It was found that the plasma concentration-time curves of the preparetion fitted two-compartment model.The main pharmacokinetic parameters were as follows: C_(max) were(28.79?3.54),(67.56?8.84) and(99.28?17.57) mg?L~(-1);T_(12?) were(1.72?0.27),(1.46?0.35) and(1.13?0.33) h;T_(12?)were(23.10?3.30),(23.95?4.72) and(24.25?4.12) h;V_d were(3.332?0.471),(2.921?0.382) and(2.921?0.622) L;CL were(0.209?0.041),(0.186?0.048) and(0.166?0.039) L?h~(-1);k_(10) were(0.063?0.012),(0.064?0.016) and(0.057?0.009) h~(-1);AUC_(0-72) were(448.68?75.06),(1015.29?225.14) and(1688.42?367.44) mg?h ?L~(-1).Conclusion The pharmacokinetics of the drug in the dosage range of 100～300 mg in human body approximately fit linear dynamic features.Compared with glycyrrhizic acid and other glycyrrhizinate salts,magnesium isoglycyrrhizinate is eliminated more slowly.This is beneficial to the treatment of chronic hepatitis.

OBJECTIVE:To evaluate the safety and tolerance of magnesium isoglycyrrhizinate(MgIG)injection in Chinese healthy volunteers,and provide safe and utility dosage scheme.METHODS:36healthy volunteers were randomly divided into4groups,each group administered respectively with single-dose intravenous infusion of100,200,300mg of MgIG and mul?ti-dose intravenous infusion of200mg of MgIG,qd for7consecutive days.Clinical symptoms,vital signs,electrocardiograms,routine blood and urine tests,as well as hepatic and renal function were observed before and after IV infusion of MgIG.RESULTS:In single-dose IV infusion of MgIG groups,the vital signs and laboratory indices did not change in the subjects immediately after or at1h,8h,24h after administration,as compared with before,while in multi-dose constant intravenous infusion group,the vital signs and laboratory indices at4days and8days after administration were in the normal range,as compared with before.CONCLUSION:Tolerance of healthy subjects to MgIG is good,and daily dosage of100mg to200mg is recommended for clinical use.

OBJECTIVE： Four different one－ week triple therapeutic schemes to eradicate Helicobacter pylori were compared by pharmacoeconomic analysis to provide a scientific method for rational assignment of our limited medical fund.METHODS： According to literature reports,892 cases of duodenal ulcer and gastritis with Helicobacter pylori were selected.These patients were randomly divided into four groups which received different one－ week triple therapeutic schemes： OMC(Omeprazole＋ Metronidazole＋ Clarithromycin),OFC(Omeprazole＋ Furazolidone＋ Clarithromycin),OFA(Omeprazole＋ Furazolidone＋ Amoxicillin)and BFC(Colloidal bismuth subcitrate＋ Furazolidone＋ Clarithromycin).The schemes were evaluated with pharmacoeconomic cost－ effectiveness analysis.RESULTS： The cost－ effectiveness ratios of OMC,OFC,OFA,BFC for gastritis or duodenal ulcer were 16.19/17.33,17.30/13.93,11.40/10.56,11.17/10.75 respectively.They changed to 16.36/17.51,17.49/14.08,8.15/7.55,11.66/11.20 by sensitivity analysis.CONCLUSION： OFA is the most effective and inexpensive one to eradicate Hp.

OBJECTIVE:Four different one-week triple therapeutic schemes to eradicate Helicobacter pylori were compared by pharmacoeconomic analysis to provide a scientific method for rational assignment of our limited medical fund.METHODS:According to literature reports,892 cases of duodenal ulcer and gastritis with Helicobacter pylori were selected.These patients were randomly divided into four groups which received different one-week triple therapeutic schemes:OMC(Omeprazole+Metronidazole+Clarithromycin),OFC(Omeprazole+Furazolidone+Clarithromycin),OFA(Omeprazole+Furazolidone+Amoxicillin)and BFC(Colloidal bismuth subcitrate+Furazolidone+Clarithromycin).The schemes were evaluated with pharmacoeconomic cost-effectiveness analysis.RESULTS:The cost-effectiveness ratios of OMC,OFC,OFA,BFC for gastritis or duodenal ulcer were 16.19/17.33,17.30/13.93,11.40/10.56,11.17/10.75 respectively.They changed to 16.36/17.51,17.49/14.08,8.15/7.55,11.66/11.20 by sensitivity analysis.CONCLUSION:OFA is the most effective and inexpensive one to era_dicate Hp.

Objective:To observe the occurrence of ADRs in a hospital and analyze its correlative factors.Meth- od:131 ADR cases collected in a hospital in 2006 were categorized statistically analyzed.Result:121 of the 131 ADR ca- ses were induced by chemical drugs,and the other 10 cases,induced by the traditional Chinese drugs.In the 121 ADR ca- ses induced by chemical drug,the most were induced by intravenous injection,which occupied 49.62%.Antimicrobial drugs were the first category of drugs to cause ADRs,totaling 77 cases from 42 drugs.ADR most commonly manifested themselves in the injury of skin and appendents,which accounted for 52.89%(64 cases).There were 10 comparatively serious ADR cases.Conclusion:The occurrence of ADRs was related to many factors,such as administration route and drug varieties.ADRs monitoring and publicizing of ADRs knowledge should be strengthened so as to lessen and avoid the occurrence of ADRs.

OBJECTIVE:To establish a RP-HPLC method for the determination of chlorogenic acid in Qingyan granules.METHODS:Waters Breeze System was used with a Hypersil BDS C18 column(5?m,4.6mm?150mm).The mobile phase was composed of methanol and 0.4%(V/V)H3PO4 water solution(20∶80,V/V),flow rate:1.0ml/min;wavelength of UV detector:327nm,column temperature 20℃.RESULTS:The calibration curve of chlorogenic acid was linear within the range of (174～1 392)ng(r=0.9 999).The average recovery was 100.10%,RSD=1.60%(n=5).CONCLUSION:This method is simple,sensitive,as well as reliable,and is available to quality control.

Objective To investigate the phenotype and the immunoregulatory function of CD8αα+TCRαβ+regulatory T cells in peripheral blood samples from mice.Methods The distribution profile and the phenotype of CD8αα+TCRαβ+regulatory T cells in C57BL/6 mice were detected by flow cytometry.The cytokines released by CD8αα+TCRαβ+regulatory T cells upon the stimulation with anti-CD3 antibody were analyzed by cytometric bead array.The in vitro immunosuppressive activity of CD8αα+TCRαβ+regulatory T cells on activated CD4+T cells was analyzed by using flow cytometry and carboxyfluorescein succinimidyl ester ( CFSE ) .An adoptive cell transfer assay was set up to evaluate the immunoprotective effects of CD8αα+TCRαβ+ regulatory T cells in a mouse model of experimental autoimmune encephalomyelitis ( EAE) .Results CD8αα+TCRαβ+regulatory T cells were detected in liver, spleen and peripheral blood samples collected from na?ve C57BL/6 mice.Compared with CD8αβ+TCRαβ+regulatory T cells, CD8αα+TCRαβ+regulatory T cells showed a memory-activated phenotype of CD25+CD122high CD44high CD62Llow CD69high NK1.1+DX5+.CD8αα+TCRαβ+regulatory T cells could produce IL-2 after 24 hours stimulation with anti-CD3 antibody, followed by producing IFN-γ, TNF-α, IL-4, IL-17A and traces of IL-6 and IL-10. In vitro, CD8αα+TCRαβ+regulatory T cells specifically suppressed the proliferation of activated CD4+T cells ( P<0.01 ).Moreover, they could delay the onset of EAE in mice and reduce clinical score (P<0.01).Conclusion CD8αα+TCRαβ+regulatory T cells were a unique population with immunoregula-tory function, which could be used as a potential therapeutic target in the treatment of autoimmune disease.