Objective To study the effects of wedelolactone on the expression of P-glycoprotein (P-gp) and to explore the multi-drug resistance reversing mechanism of wedelolactone in K562/A02 cells in vitro.Methods The half maximal inhibitory concentration of ADM in K562/A02 was determined by MTT method.The protein expression level of P-gp was determined by Western blot after wedelolactone pretreatment.Results Wedelolactone remarkably enhanced chemo-sensitivity to ADM of K562/A02 cells.After 0.2, 2, 20 μmol/L different concentration of wedelolactone treatment in 24 h, the relative reversal efficiency of K562/A02 to ADM was 23.5％, 47.1％ and 67.7％, respectively.according to the results of Western blot, wedelolactone was shown to efficiently inhibit the expression of P-gp (P＜0.05).The relative efficiency of K562/A02 to ADM was 25.4％,46％ and 55.6％, respectively.Conclusion Wedelolactone could modulate P-gp expression, and P-gp expression down regulation may be one of the MDR reversal mechanisms in K562/A02 cells by wedelolactone.

Objective To study the influence of HBV replication regulator,enhancer I and Pre- S2,on the immune response of HBV DNA vaccine.Methods DNA fragments of HBsAg,PreS2 HBsAg,HBsAg-enhancer I and PreS2-HBsAg-enhancer I region of HBV were amplified by PCR using the complete genome DNA of HBV adr subtype,and inserted into VR1012 vectors,respective- ly.The recombinant plasmids were transfected into HepG2 cells,and injected into Balb/C mice.The expression of HepG2 cells and the cellular and humoral immune response of mice were tested by cell immuno-chemistry,ELISA and ELISPOT.Results The target protein were expressed by transfected HepG2 cells,enhancer I and Pre-S2 can promote the expression of HBsAg in transfected cells.The HBsAb and the HBsAg specific CTL in inoculated mice were found in the second week after injection, PreS2 but not enhancer I can promote the immune response in inoculated mice.Conclusions When inserted into HBV DNA vaccine,enhancer I and PreS2 can promote the expression of HBsAg in transfected HepG2 cells,PreS2 can promote the immune response in inoculated Balb/C mice.

AIM To establish a quantitative analysis of multi-components by single marker(QAMS) method for the content determination of six catechins in Xinnaojian Capsules (Tablets) (tea extract).METHODS The analysis of 50％ methanol extract of this drug was performed on a 35 ℃ thermostatic Shimadzu Wonda Cract ODS-2 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of 0.5％ acetic acid (A)-acetonitrile (B) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 280 nm.With epigallocatechin gallate as an internal standard,the relative correction factors of epigallocatechin,catechin,epicatechin,gallocatechin gallate and epicatechin gallate were calculated,from which the content determination was made.RESULTS Six constituents showed good linear relationships within their own ranges (r ≥ 0.999 8),whose average recoveries were 96.00％-98.47％ with the RSDs of 2.09％-2.91％.The results obtained by QAMS method approximated those obtained by external standard method.CONCLUSION This simple and reliable method can be used for the quality control of Xinnaojian Capsules (Tablets).

BACKGROUND:Many studies reported the relationship of the mechanical properties and water content about bone tissue, which is one of organizations containing the lowest water content on human body. Researches on effect of water on biological tribology behavior of bone tissue have been rarely reported and are the experimental study generally. OBJECTIVE:To explore the influence and the damage mechanism of water on biological tribology behavior of bone tissue, by comparing multiscale numerical model established with the experiment. METHODS:Dehydration of the bone tissue was studied by nanoindentation test and both reciprocating sliding and impact wear tests. A multi-scale finite element model was constructed under a flat-on-ball configuration. RESULTS AND CONCLUSION:The viscoelasticity and the tribological properties of bone tissue significantly decreased as well as the different wear mechanisms under applied loading after drying. The analytical results indicated that there were high stress condition, which incurred the micro-crack initiation and the appearance of peeling and wear, around the Haversian canal, circumferential lamellas and the interstitial tissues. Meso-scale:dehydration weakened the function of absorption and interruption of stress, which facilitated crack extension in pore. Micro-scale:the high stress gradient of structure of canaliculi and lacunae is an important cause of tissue damage.

Objective To analyze the differentially expressed genes between the NCAM + c-Kit +RBE and NCAM-c-Kit-RBE of intrahepatic cholangiocarcinoma (ICC) cell lines,and to screen out the differentially expressed genes that are related to the stem cell signaling pathways.Methods Magnetic activated cell sorting was used to isolate the NCAM + c-Kit +/NCAM-c-Kit-subset cells,and then Agilent Whole Human Genome Microarray Kit was used to test the difference in gene expressions between the NCAM + cKit + and NCAM-c-Kit-subset cells.The difference in gene expressions related to the stem cell signaling pathways was analyzed by the SAS system.The result of the microarray was further confirmed by RT-PCR.Results The total differentially expressed genes which could be found through gene microarray were 7270 [foldchange(fc) ≥2 or fc ≤0.5].Compared with the NCAM-c-Kit-RBE,3572 genes were upregulated while 3698 genes were downregulated.The differences in gene expressions related to the stem cell signaling pathways were 421 (fc ≥2 or fc ≤ 0.5),among which 231 genes were upregulated while 190 genes were downregulated.Conclusions High-flux microarray could be used to screen out lots of differentially expressed genes between the NCAM + c-Kit + and NCAM-c-Kit-RBE cells.The differences in gene expression in the stem cell signaling pathways could also be further analyzed using the SAS system.

AIM:To observe whether modified epitopes from hepatocellular carcinoma antigen MAGEC 2 have HLA-A2-restricted antitumor ability.METHODS:HLA-A2 epitopes from MAGEC2 protein were predicted by NetCTL 1. 2,SYFPEITHI and IEDB.The change of binding anchor motifs by replacing anchor residues created the modified peptides from MAGEC2.The binding affinity of the peptides to HLA-A*0201 molecule was evaluated by T2 cells binding assay. ELISPOT assay and intracellular cytokine staining were used to investigate the ability of the peptides inducing specific re -stricted cytotoxic T-lymphocytes(CTLs)to release interferon-γ(IFN-γ).The ability of the peptides to induce T-cell re-sponse was investigated by cytotoxicity assay in vitro.RESULTS:The candidate peptides P248, P248-1Y,P356,P356-1Y,P356-2L and P356-1Y2L showed moderate affinity toward HLA-A2 molecule.T2 binding assay showed that P248-1Y and P356-1Y2L showed significantly higher affinity for HLA-A2 than the native peptides.ELISPOT assay and intracellular cytokine staining showed P248, P248-1Y, P356 and P356-1Y2L were able to induce specific CTLs to release IFN-γ. ELISPOT assay showed that significantly higher levels of IFN-γrelease were induced by P248-1Y and P356-1Y2L than the native peptides.The CTLs induced by P248,P248-1Y,P356 and P356-1Y2L lysed HepG2 cells,and P248-1Y and P356-1Y2L peptide-specific CTLs showed higher cytotoxicity against HepG 2 cells than the native peptide-specific CTLs(P<0.05).CONCLUSION: Compared with the native peptides, modified epitopes P248-1Y and P356-1Y2L have higher binding affinity with HLA-A2 and retain immunogenecity.In addition, the antitumor immunity effects of modified epitope P248-1Y and P356-1Y2L are stronger than the native peptides.The peptides P248-1Y and P356-1Y2L are excellent HLA-A2-restricted CTL epitopes from tumor antigen MAGEC 2, which could serve as new candidates towards antitumor peptide vaccines.

Objective To compare the effects of different methods of general anesthesia on postoperative cognitive function in patients undergoing non-cardiac surgery.Methods One thousand ASA Ⅰ or Ⅱ patients,aged 18-60 years and undergoing non-cardiac surgery,were randomly divided into five groups (n=200 each):isoflurane + propofol + fentanyl group (group IPF),isoflurane + remifentanil group (group IR),sevoflurane + propofol + fentanyl group (group SPF),sevoflurane + remifentanil group (group SR),and propofol + remifentanil group (group PR).Two hundred patients receiving non-operative treatment served as control group (group C).In groups IPF and SPF,anesthesia was maintained with inhalation of 1.68％ isoflurane or 1.71％ sevoflurane,target controlled infusion (TCI) of propofol with the target plasma concentration of 2-5 μg/ml,and intermittent intravenous boluses of fentanyl.In groups IR,SR and PR,anesthesia was maintained with inhalation of 1.68％ isoflurane or 1.71 ％ sevoflurane,or TCI of propofol with the target plasma concentration of 2-5 μg/ml,and TCI of remifentanil with the target plasma concentration of 2-6 ng/ml.The patients' cognitive function was assessed with minimental state examination (MMSE) 1 day before operation,when leaving the post-anesthetic care unit (PACU),and 1 and 3 days after operation,respectively.Z score was used to identify the cognitive dysfunction as recommended by Moiler when leaving the PACU,and 1 and 3 days after operation.Results Compared with group C,the MMSE score was significantly decreased when leaving the PACU,and the incidence of cognitive dysfunction increased when leaving the PACU and 1 day after operation in the other groups (P ＜ 0.05).Compared with groups IPF,IR,SPF and PR,the incidence of cognitive dysfunction was significantly increased in group SR (P＜0.05).Conclusion General anesthesia with sevoflurane combined remifentanil exerts fewer effects on the postoperative cognitive function in patients undergoing non-cardiac surgery.

The fine structure of enoxaparin sodium samples with different degree of 1,6-anhydro derivatives were analyzed with polyacrylamide gel electrophoresis, high performance liquid chromatography, ultraviolet spectroscopy, infrared spectroscopy and nuclear magnetic resonance spectroscopy. A further study of anticoagulation activity of enoxaparins was performed, including those on their inhibition activities of coagulation factor Xa (FXa) and thrombin (FIIa). The results showed that the anti-FXa and -FIIa activities of enoxaparins with different degree of 1,6-anhydro derivatives (20.0%-39.7%) with similar structure characteristics, had decreasing tendency when the degree of 1,6-anhydro derivatives increased. Especially, the anti-FXa activity was sensitive to the change of the degree of 1,6-anhydro derivatives.
Anticoagulants ; chemistry ; Enoxaparin ; chemistry ; Factor Xa Inhibitors ; chemistry ; Thrombin ; antagonists & inhibitors

Objective To screen thyroid function among planned pregnant women in Chongqing, to guide prenatal and postnatal care. Methods Proportional multi-stage stratified cluster random sampling method was adopted to enroll 11 953 planned pregnant women for questionnaire, physical examination, and serum samples collection. Results The median TSH was 2. 04 mIU/ L, P25 = 1. 36 mIU/ L,P75 = 2. 99 mIU/ L. TSH levels being normal, higher, and lower than the reference were 91. 47% , 6. 20% , and 2. 33% , respectively. In Northeast Chongqing, the proportions of median TSH level and TSH level above the upper limit were higher than those in other regions(P<0. 05). With improved education, proportions of TSH above the upper limit and below the lower limit declined(P<0. 05). With the increase in body mass index, the proportion of those whose TSH was above the upper limit showed elevated trend(P<0. 05). In women with history of adverse pregnancy outcomes, their median TSH was higher than that in the control group, and those, whose TSH level exceeded the upper limit, yield higher results than those in the control group(P<0. 05). In women with higher fasting blood glucose The median TSH level was lower than that in normal blood glucose group( P<0. 05), with the fasting plasma glucose concentration and TSH negatively correlated(P<0. 05). Conclusion The abnormal rate of TSH level in planned pregnant women was 8. 53% in Chongqing. The abnormal rate varies by different regions, education levels, body mass indexes, and blood glucose levels. Previous history of adverse pregnancy outcomes was related to elevated TSH levels. It is necessary to take pre-pregnancy thyroid function screening investigation.

Objective To investigate the effect of clotrimazole on apoptosis of hepatic cells after ischemia-reperfusion injury and its mechanism. Methods Hepatic ischemia-reperfusion rat model was established. Thirty-two male Sprague-Dawley rats were randomly allocated into sham-operated group, model control group, low dose clotrimazole group and high dose clotrimazole group. Apoptosis in hepatic tissue was assessed by TUNEL method. Protein expression levels of CYP3A1,Bcl-2,Bax and PARP were measured by Western blotting. Results As compared with model control group, the apoptosis rate, tissue injury,activity of plasma enzymes and the Bax/Bcl-2 expression ratio were reduced in low and high dose clotrimazole groups. The apoptotic index in both clotrimazole-treated groups was lower than that of model control group with statistically significant difference. CYP3A1 expression was significantly induced by clotrimazole compared to the sham-operated group. Conclusion Clotrimazole may inhibit apoptosis of hepatic cells by up-regulating Bcl-2 and down-regulating Bax, thus produce a protective effect on hepatic ischemia-reperfusion injury and it is also related to the inhibition of PARP shear.