AIM: To observe the direct effect of lipopolysaccharide (LPS) on secretion of endothelin-1(ET-1) and nitric oxide by human umbilical vein endothelial cell and cell viability of the secretor. METHODS: The third passage of human umbilical vein endothelial cells were incubated with different concentrations of LPS(1 g/L, 100 mg/L, 10 mg/L, 1 mg/L, 100 ?g/L, 10 ?g/L, 1 ?g/L) for 6 hours, and the culture supernatants were collected. The concentrations of ET-1 were determined by radioimmunoassay, the concentrations of nitric oxide were determined using Greiss's method. The viabilities of cells were measured by MTT method. RESULTS: The concentration of ET-1 (pg/L) of normal control group was 251 64?10 90. The concentrations of ET-1(pg/L) of LPS treated groups were 220 85?19 14, 278 67?15 45, 306 40?11 60, 312 87?33 50, 324 38?17 02, 291 49?14 30, 282 11?13 38, respectively. (each group compared with normal control group, P

Objective To observe the effect of Xinmailing Injection (XI) on lipopolysaccharide(LPS)- induced secretion of endothelin- 1 (ET- 1) and nitric oxide (NO) from endothelial cells of human umbilical veins.Methods The in- vitro culture of human umbilical vein endothelial cells were stimulated by LPS( 100 ? g/L) and incubated with XI(320 g /L) for half an hour,one hour,2 hours,4 and 6 hours respectively.Then the cultured supernatants were collected.The viabilities of cells were measured by MTT method.The concentrations of ET- 1 and NO were determined by radioimmunoassay method and by Gneiss's Method respectively.Results (1) The viabilities of cells arrived the highest level as treated with XI with the concentration of 320 g /L (P

AIM and METHODS: To investigate the effect of endotoxin on the celluar activity and secretion of endothelin-1 by radioimmunoassay and MTT methods in cultured human umbilical vein endothelial cells stimulated by E coli endotoxin (E coli O55:B5, Sigma) of various concentrations (1 g/L, 100 mg/L， 10 mg/L，1 mg/L，100 μg/L，10 μg/L, 1 μg/L) and at the same time interval (HUVEC stimulated by endotoxin for 6 hours) in vitro.RESULTS:Endotoxin showed a slightly inhibitory effect on the viability of endothelial cells at low doses (1 μg/L， 10 μg/L, 100 μg/L, 1 mg/L). The viabilities were 92.00%±1.45%， 91.81%±2.03%, 89.52%±1.49%, 88.35%±1.88%, respectively, versus control group, P＜0.01. The cells were impaired significantly at the higher dose of LPS (100 mg/L), the viability was 80.49%±8.76%, versus control group, P＜0.01. The cells were killed evidently at the concentration of LPS (1 g/L), the viability was 73%±8%, versus control group, P＜0.01. The secretion of ET-1 increased gradually with the concentration of endotoxin manifolding. The concentration of ET-1 reached its peak at the dose of 100 μg/L, and it was (324.384±17.023) ng/L, versus control group (251.636±17.023) ng/L, P＜0.01. Endotoxin was effective in stimulating the endothelial cells to secret ET-1 in a dose dependent manner. CONCLUSION: These findings suggested ET-1 may be one of the important factors in endotoxic shock, and the increase in plasma ET-1 level in endotoxemia may be associated with increase in ET-1 secretion.

To study the direct effect of E.Coli endotoxin on the production of nitric oxide by endothelial cells, the second passage of cultured human umbilical cells was stimulated by serial doses of endotoxin (1 g/L, 10 mg/L, 100 μg/L, 10 μg/L, 1 μg/L), and the content of nitric oxide in supematant of culture and the viability of endothelial cells 6 hours after the stimulation were obcerved. The result showed that endotoxin had a slightly inhibitory effect on both the production of nitric oxide and the viability of endothelial cells at low doses (1 μg/L, 10 μg/L, 100 μg/L), especially the dose of 100 μg/L ［(608.63±11.64) μmol/L, versus that of unstimulated grouop (629.46±13.36) μmol/L, P＜0.05］. While the high doses of endotoxin exerted a big increasing in production of nitric oxide and a big decrease in the viability of endothelial cells, especially the dose of 1 g/L (NO: 722.58 μmol/L±32.18 μmol/L, versus that of unstimulated group P＜0.01; viability: 73.63%±8.50%, versus that of unstimulated group, P＜0.01). These could be concluded that low doses of endotoxin mainly resulted in functional changes in endothelial cells, such as decrease in relaxing factor (nitrc oxide), while high doses endotoxin exerted lethal effects on endothelial cells accompanied with high production of nitric oxide, which might be related to the death of cells.

AIM: To observe the direct effect of LPS on expressions of ET-1, eNOS, and iNOS mRNA in human umbilical vein endothelial cells, and further research the molecular mechanism of effect of LPS on production of ET-1 and NO. METHODS: The third passage of cultured human umbilical vein endothelial cells was incubated with low concentration (100 ?g/L) of LPS for 6 hours. Total RNA was extracted. The expressions of ET-1, eNOS, and iNOS mRNA were analyzed by semi-quantitative RT-PCR method. RESULTS: ET-1 mRNA experession increased significantly, while expression of eNOS mRNA decreased significantly, and there was no significant change in expression of iNOS mRNA. CONCLUSIONS: In human umbilical vein endothelial cells, low concentration of LPS enhanced the expression of ET-1 mRNA, inhibited the expression of eNOS mRNA, and had no significant effect on the expression of iNOS mRNA.

Objective To study the influence of bFGF gene transfected bone marrow-derived mesenchymal stem cells (BMSCs) on the inflammatory cytokines of COPD rat. Methods The BMSCs were separated from SD rat and cultured and then bFGF gene was imported to BMSCs by liposome transfection method. The samples were prepared into six groups: normal control group, COPD group (A), BMSCs group (B), pcDNA3.1-BMSCs group (C), bFGF-pcDNA3.1-BMSCs group (D), and bFGF group (E). The expressions of TNF-α and IL-1β by QRT-PCR were detected. Results Compared with COPD group, TNF-α and IL-1β genes from groups B to D dropped significantly (P < 0.05). The changes of TNF-α and IL-1β genes among groups B to D showed no significant difference (P > 0.05). Conclusion BFGF transfected BMSCs, sample BMSCs and pcDNA3.1 transfected BMSCs can inhibit the expression of inflammatory cytokines of TNF-α and IL-1β, but there is no obvious advantage in comparison to bFGF transfected BMSCs and sample BMSCs in respect of inhibiting the expression of inflammatory cytokines of TNF-α and IL-1β.

Objective Using quantitative real-time PCR to establish a rapid specific genetic diagnostic technique for Yersinia pestis.Methods ①Four sets of specific probes and primers were designed,which targeted to chromosome genes of YPO0392,YPO1094,YPO2087 and YPO2090,respectively.②The probes and primers were tested for stability and specificity with 40 strains of Yersinia pestis and 47 strains of non-Yersinia pestis of different sources in Yunnan.③Eight positive DNA in Yulong,Yunnan,were tested with the screened probes and primers.Results ①Two sets probes and primers were selected,they were targeting YPO0392 and YPO1094,respectively.②The results were all positive of the eight positive DNA samples tested.Conclusion Two sets of primers and probes are selected for rapid specific diagnosis of Yersinia pestis.

Objective Bone marrow mesenchymal stem cells ( BMSCs) , a kind of stem cells with multiple differentiation po-tentials, exist in the bone marrow and other organizations.This study aimed to investigate the repairing effect of the exogenous basic fi-broblast growth factor ( bFGF) against chronic obstructive pulmonary disease ( COPD) and its action mechanism, and to determine the expression of the bFGF gene in transfected rat BMSCs. Methods BMSCs were isolated, cultured and identified.The recombinant plasmid bFGF-pcDNA3.1 was constructed and sequenced.Liposome-mediated bFGF-pcDNA3.1 plasmid was transfected into the BM-SCs of the rat (bFGF-pcDNA3.1 transfection group), liposome-mediated pcDNA3.1 transfected into the BMSCs (pcDNA3.1 transfec-tion group) , and untransfected BMSCs used as the control.G418 screening was performed for 14 days.The gene and protein expres-sions of bFGF were determined by qRT-PCR and Western blot. Results The full-length sequence of the bFGF gene was consistent with that of the GenBank.The expression of the bFGF gene was significantly higher in the bFGF-pcDNA3.1 transfection group (7.028 ±0.568) than in the pcDNA3.1 transfection group (1.000 ±0.082) and the non-transfection control (1) (P<0.01), but with no statistically significant difference between the latter two groups (P>0.05).The expression of the bFGF protein was also re-markably higher in the bFGF-pcDNA3.1 transfection group (1.017 ±0.054) than in the pcDNA3.1 transfection group (0.217 ± 0.009) and the non-transfection control (0.165 ±0.013) (P<0.05), with no statistically significant difference between the latter two groups (P>0.05). Conclusion Mediated by the liposome reagent, the recombinant eukaryotic expression vector bFGF-pcD-NA3.1 can be transfected into rat BMSCs and expresses the bFGF gene and protein.

OBJECTIVE:To discuss the characteristics and regular patterns of adverse drug reactions (ADRs) induced by Qingkailing injection for references of clinical rational drug use. METHODS:ADR cases induced by Qingkailing injection reported in Chinese pharmaceutical journals from 2003 to 2005 were retrieved and analyzed statistically. RESULTS:The ADRs induced by Qingkailing injection were more often seen in men than in women, with children and young adults showing higher percentages. Allergic reactions were the main type of adverse drug reactions. Of the total ADR cases induced by Qingkailing injection, 22 cases(33.33%) were anaphylactic shock,15(22.73%) were allergic reactions,10(15.15%) were allergy of circulation system. CONCLUSION:The ADRs induced by Qingkailing injection may be resulted form many factors, which should be given fully attention in the clinic. ADRs monitoring should be stressed when this drug being used by western medicine physicians to make sure rational use of which and to reduce incidences of ADRs.

OBJECTIVE:To investigate the situation and characteristics of serious adverse drug reactions (ADR) occurred in our hospital in order to promote rational clinical drug use. METHODS: 25 severe ADR cases collected in our hospital from Jun. 2004 to Dec. 2006 were analyzed statistically and evaluated based on the ADR causality judgment criteria. RESULTS: The 25 severe ADR were associated with 21 kinds of drugs, with anti-infectives (12 kinds) making up the highest proportion, followed by traditional Chinese medicine preparations. The main systems involved in he ADR were skin and its appendages and nervous system. 3 new and severe ADR cases were found to have no report in package inserts or literature. CONCLUSION: Severe ADR may be induced by many factors, therefore, it is necessary to strengthen the awareness of rational drug use to guard against severe ADR and ensure safe and effective medication.