Objective:To develop an HPLC method for the determination of capsaicine, dihydrocapsaicin, imperatorin and isoim-peratorin in Guanjie Jietong plasters. Methods:A CAPCELL PAK C18 column was used as the chromatographic column, and the flow rate was 0. 8 ml·min-1. The mobile phase A consisted of methanol-acetonitrile(1∶1), the mobile phase B was of 1% phosphoric acid. The detection wavelength was 280 nm and 254 nm, respectively. Results:There was a good linear relationship between the con-centrations and the peak areas within the range of 0. 298-5. 956μg(r=0. 9998) for capsaicine, 0. 152-3. 044μg(r=0. 999 6) for di-hydrocapsaicin, 0. 018-0. 352 μg(r=0. 999 5) for imperatorin and 0. 010-0. 204 μg(r=0. 999 3) for isoimperatorin. The average re-covery was 97. 8%(RSD=1. 02%), 97. 0%(RSD=0. 76%), 96. 6%(RSD=0. 65%) and 98. 1%(RSD=1. 35%), respectively. Conclusion:The method is convenient, accurate, sensitive and repeatable, which can be used in the determination of capsaicine, di-hydrocapsaicin, imperatorin and isoimperatorin in Guanjie Jietong plasters.

Objective:To investigate the impact of recombinant flagellin targeting TLR5 and NLRC4 simultaneously or respectively on innate immune cells in mice. Methods: Induction,expression,purification and identification of recombiant FliC,which were FliC(activating both TLR5 and NLRC4);FliCΔ90-97(unable to activate TLR5),FliC-L3A(unable to activate NLRC4),FliCΔ90-97:L3A(unable to activate both TLR5 and NLRC4). The mice were divided into five groups,namely group FliC,FliC-L3A,FliCΔ90-97,FliCΔ90-97:L3A and PBS,which were injected with 100μl PBS or 10μg recombinant flagellin intraperitoneally,three mice in each group. 12 h later,the mice were executed using dislocation of cervical vertebra and the splenic and peritoneal cells were isolated. The spleen was grinded into single-cell suspension. The proportion of neutrophils,NK cells,DCs and the expression level of CD80 and CD86 on DCs were evaluated with flow cytometry. Results:Group FliC,group FliC-L3A and group FliCΔ90-97 shared the similar proportion of neutrophils in peritoneal cavity ( P>0. 05 ) , and all of which were significantly higher than group PBS and group FliCΔ90-97 ( P<0. 01),and NK cells also showed the similar trend. Compared with group FliCΔ90-97 and FliCΔ90-97:L3A,the mean fluorescence intensities(MFIs) of CD80 and CD86 in group FliC and FliC-L3A increased significantly(P<0. 01). The proportion of Treg in spleen was highest among all groups. Conclusion:Activation of TLR5 and NLRC4 had similar chemotaxis of neutrophils and NK cells. The ex-pression of CD80 and CD86 on DCs were upregulated after stimulation by flagellin and TLR5-dependent. Activation of TLR5,but not NLRC4,increased the proportion of Treg in spleen.

Objective:To explore the impact of TLR5 and NLRC4 activation on the proliferation of different breast cancer cell lines,MCF-7 and MDA-MB-23 i.Methods:Induction,expression,purification and identification of recombiant flagellin,including FliC (activating both TLR5 and NLRC4),FliC△90-97 (unable to activate TLR5),FliC-L3A (unable to activate NLRC4),FliC△90-97:L3A (unable to activate both TLR5 and NLRC4).Using different concentration of recombinant flagellin to stimulate MCF-7 and MDA-MB-231 cell lines,72 h later,the proliferation of tumor cells were detected with CCK8.We also used soft AGAR forming experiments to detect the inhibition ratio of recombinant flagellin on breast cancer cell lines.Briefly,1 000 cells were plated in the 6-well plate,then stimulated with 1 μg/ml recombinant flagellin,14 days later,the number of cloning were counted after crystal violet staining.Results:After stimulation with four recombinant flagellins at the concentration of 0.1 μ,g/ml,the inhibition ratio on MCF-7 reached 30％,and FliC△90-97 were dose-dependent on the inhibition of MCF-7 proliferation.At the concentration of 1 μg/ml,FliC-L3A which only activated TLR5 showed stronger inhibition ratio than FliC.FliC△90-97:L3A which did not activate both TLR5 and NLRC4 also inhibited the proliferation of MCF-7.After adding transfection reagent,four recombinant flagellins showed inhibition effect on MDA-MB-231.Conclusion:Flagellin can inhibit the proliferation of MCF-7 and MDA-MB-231,and the mechanism of inhibition on the proliferation were not TLR5 and NLRC4 pathway dependent.There might exist new mechanisms to explain this phenomenon.

1000 ml).Methods From 2000 to 2002,a total of 89 critically ill surgical patients were enrolled whose preoperative liver function and blood analysis were normal. The patients′ heart rate?blood pressure?body temperature?hemoglobin(Hb)?platelet count(PLT)?hematocrit(HCT)?prothrombin time(PT)?activated partial thrombo-plastin time(APTT)?fibrinogen concentration(FIB)?fibrinogen degrade product(FDP)?D-dimer immediately after enrollment into the study were determined. Results As the blood loss increased PT prolonged from(11.4?0.6) s to (16.0?0.6) s ( P

Objective To investigate the effects of liver X receptor agonist on the expressions of C-reactive protein and CD40 ligand and smooth muscle cell ?-actin in the aorta of ApoE gene knockout mice with earlier atherosclerosis. Methods Male ApoE gene knockout mice (8-week old) were divided randomly into control group and T0901317 treatment group (n=6 in each group). The mice in T0901317 group were administered intraperitoneally with T0901317 at the dose of 20 mg?kg-1?d-1 for 4 weeks. Mice in the control group were only given equivalent amount of dimethyl sulfoxide (DMSO). The expressions of C-reactive protein and CD40 ligand and smooth muscle cell ?-actin were detected by immunological histochemical method. Results The expressions of C-reactive protein and CD40 ligand in the atherosclerotic plaque in the aortic wall were significantly lower in T0901317 group as compared with those in the DMSO control group (P0.05). Conclusion Liver X receptor agonist may reduce the formation of atherosclerotic lesions by inhibiting the inflammation and the expressions of C-reactive protein and CD40 ligand in the aorta of ApoE gene knockout mice.

OBJECTIVE:To improve the preparation of compound menthol spray and study its stability.METHODS:The liquor compound menthol spray was changed to emulsified ones by the solubilizing action of tween80.The stability of emulsified compound menthol spray(ECMS)was observed in airtight container at room temperature(25?2)℃.RESULTS:ECMS was stable within2wks in airtight container at room temperature.CONCLUSION:ECMS is stable in property,convenient in stor?age,and accords with pharmaceutical quality standards.

100 U/ml and treated for 48 h,the inhibitory ratio of the HepG2 cells was raised to (39.08?2.32)%,which were not further elevated with the increase of dose or elapse of treatment time. HepG2 cells apoptosis was induced by thrombin.in a dose-and time-dependent manner when the dose was between 25 to 100 U/ml. However,there was no significant difference in the apoptosis of different groups when the dose was higher than 100 U/ml and the time exceed 48 h. Conclusion Higher doses of thrombin can inhibit the proliferation of HepG2 cells,and lead to cell apoptosis. The both effects are increased in a dose-and time-dependent manner when the dose is

Objective To investigate the effects of celastrol on the expressions of macrophage migration inhibitory factor(MIF)and matrix metalloproteinase-9(MMP-9)in the aorta of apoE gene knockout mice with earlier atherosclerosis.Methods Eight-week-old ApoE gene knockout male mice were divided randomly into control group and celastrol treatment group(n=6 in each group).The mice in celastrol group were given.celastrol(2 mg?kg-1?d-1)by intraperitoneal injection for 4 weeks;and the mice in control group were only given equivalent amount of dimethyl sulfoxide(DMSO).HE staining of root aorta were used to observe the histomorphological changes and measure the size of plaque in ApoE-/-mice.The expressions of MIF and MMP-9 were detected by immunological histochemical method.Results The area of lipid plaque in the mice treated with celastrol was significantly smaller than that of the control(P

BACKGROUND:Currently, human umbilical cord derived-mesenchymal stem cel s are mainly for local transplantation, which has some shortcomings, such as large trauma, bleeding, complications, that limit its widespread application in clinical practice. OBJECTIVE:To investigate the feasibility of intravenous transplantation of human umbilical cord derived-mesenchymal stem cel s for repair of spinal cord injury. METHODS:Eighty Wistar rats with spinal cord hitting were divided into five groups:blank control group with no transplantation (n=10), DMEM local transplantation group (n=15), DMEM intravenous transplantation group (n=15), cel local transplantation group (n=20), cel intravenous transplantation group (n=20). The functional recovery of spinal cord injury was observed with Basso, Beattie and Bresnahan scores at regular time as wel as hematoxylin-eosin staining and immunohistochemistry staining. RESULTS AND CONCLUSION:During 1 day to 2 weeks after transplantation, there was no significant difference in the Basso, Beattie and Bresnahan scores between the five groups;within 4-12 weeks after transplantation, the Basso, Beattie and Bresnahan scores were significantly higher in the two cel transplantation groups than the other three groups, but there was no difference between these two cel transplantation groups (P>0.05). Histological observation showed that the number of voids and glial scars was less in the cel local transplantation group and cel intravenous transplantation group compared with the other three groups, and there was also no difference between the two cel transplantation groups. These results indicate that the intravenous transplantation of human umbilical cord derived-mesenchymal stem cel s is similar to the local transplantation in the repair of acute spinal cord injury, which is simple and avoids secondary injuries and various complications. It is recommended that this method provide a new approach for cel transplantation.

Objective To explore the relationship between the expression of human telomerase RNA component (TERC) gene , human papilloma virus (HPV) infection and mutation of chromosome 3 number with cervical lesions .Methods 81 women received the treatment in the Gynecology Department of the Second Affiliated Hospital of Kunming Medical University from June 2008 to February 2009 ,including the healthy group(normal pathological examination ,20 cases) ,CIN1 group(28 cases) ,CIN2 group(12 ca‐ses) ,CIN3 group(9 cases) and cervical cancer group(12 cases) .The TERC gene expression in uterine epithelial exfoliated cells was detected by using the fluorescence in situ hybridization (FISH) method ,meanwhile the HPV infection was detected by using the real time fluorescence quantitative polymerase chain reaction (FQPCR) technology .The correlation between cervical cancer with TERC gene and HPV was analyzed .At the same time the number of chromosome 3 mutations in 81 cases was recorded .Results In the cervical lesion detection ,the detection positive rate had no statistical difference between the TERC gene detection and HPV detec ‐tion (P> 0 .05) ,their positive rates in the CIN 1 ,CIN2 ,CIN3 and cervical cancer groups were significantly higher than that in the healthy group (P< 0 .05) ,the difference between the CIN1 group and the CIN2 group had no statistical significance (P > 0 .05) , while between the CIN3 group and the cervical cancer group had statistical significance (P< 0 .05) ,the higher the malignant degree , the higher the positive rate .The abnormal mutation rate of chromosome 3 number was 0% in the healthy group and the CIN1 group ,16 .7% in the CIN2 group ,66 .7% in the CIN3 group and 100 .0% in the cervical cancer group ,the positive rate in the CIN3 group and the cervical cancer group was significantly higher than that in the healthy group ,CIN1 group and CIN2 group ,the differ‐ences were statistically significant (P< 0 .05) .Conclusion The TERC abnormal gene expression ,high risk HPV infection and mutation of chromosome 3 number could play an important synergistic effect during the process of occurrence and progression of cervical cancer .