ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Fuzhenghuayu compound (FZHc) on expression of nuclear factor E2-related factor 2 (Nrf2) in hepatocytes under conditions of hepatic fibrosis using a mouse model.</p><p><b>METHODS</b>Mice were randomly assigned to a control group and a hepatic fibrosis model group. The control group was further divided into three subgroups for use as normal controls (A1), mineral oil-treated controls (A2), and FZHc-treated controls (A3); the hepatic fibrosis model group was administered carbon tetrachloride (CC14 dissolved in mineral oil and injected intraperitoneally) and further divided into four subgroups for use as 6-weeks models (B1), 10-weeks models (B2), low-dose (L)-FZHc models (C1), and high-dose (H)-FZHc models (C2). The FZHc (capsule powder diluted with double-distilled water to 0.1 g/mL) was administered via gastric perfusion to groups A3, C1, and C2 starting at week 7 of the experiment. At the end of week 6 and 10, hepatic specimens were collected and evaluated for degree of hepatic fibrosis and inflammation using routine haematoxylin-eosin staining and Masson staining. Immunohistochemical analysis was performed to measure the hepatocyte expression of Nrf2, NAD(P)H quinine oxidoreductase 1 (Nqol), a-smooth muscle actin (a-SMA) and fibronectin (FN). Real-time fluorescence quantitative PCR was used to measure Nrf2 mRNA expression. Western blotting was used to detect Nrf2 and Nqol total protein expression and Nrf2 nuclear translocation. F test, LSD test and ridit test were used for statistical analyses.</p><p><b>RESULTS</b>Compared with the B2 group (ridit value: 0.09), the model groups treated with FZHc showed significantly lower degrees of hepatic inflammation and fibrosis for both the low (C1 group, ridit value: 0.32) and high doses (C2 group, ridit value: 0.40) (F =82.927, P less than 0.05). In addition, compared with the B2 group, the model groups treated with FZHc showed significantly decreased expression of a-SMA and FN proteins, with a dose-dependent trend (by immunohistochemistry: C 1 group at the end of 10 weeks, F =77.421, 118.262, P less than 0.05; C2 group, P =0.002, 0.013) and significantly increased expression of Nrf2 and Nqol proteins (by immunohistochemistry:C1 and C2 groups at the end of 10 weeks, F =182.537, 75.615, P less than 0.05 and by westen blotting: F =45.664, 127.673, P less than 0.05), which also showed a dose-dependent trend (C2 group, P =0.000, 0.014; 0.005, 0.014). Western blotting also indicated that the amount of nuclear transported Nrf2 was higher in the C1 and C2 groups at the end of 10 weeks (vs. B2 group, F =94.787, P less than 0.05), and the amount of nuclear transported Nrf2 was significantly higher in the C2 group (vs. C1 group, P =0.044). Nrf2 mRNA expression was significantly higher in the C1 group than in the B2 group (F =3230.105, P less than 0.05), and the C2 group had more substantially increased expression (P =0.001); there was no statistical difference found between groups B1 and B2 (P =0.094).</p><p><b>CONCLUSION</b>Fuzhenghuayu compound increased the expression of Nrf2 mRNA and protein under conditions of hepatic fibrosis in mice and stimulated Nrf2 nuclear transport, as well as increased expression of the Nrf2 target gene Nqol that is known to suppress activation of hepatic stellate cells and decrease the deposition of FN. Therefore, Fuzhenghuayu compound may ameliorate hepatocyte injury in hepatic fibrosis in mice by exerting an antihepatic fibrosis effect.</p>
Subject(s)
Animals , Female , Mice , Drugs, Chinese Herbal , Pharmacology , Hepatocytes , Metabolism , Liver Cirrhosis, Experimental , Metabolism , Mice, Inbred Strains , NAD(P)H Dehydrogenase (Quinone) , Metabolism , NF-E2-Related Factor 2 , MetabolismABSTRACT
ObjectiveTo analyze the status of DACH1 gene promoter methylation and explore its association with the expression of DACH1 gene promoter methylation and clinical significance of endometrium carcinoma(EC).Methods From February 2004 to August 2008,a total of 80 EC tissue samples with comprehensive surgical pathology staging were collected and used for this study.Twenty normal endometrium tissues in 2008 were abtained from the fractional curettage because of dysfunctional uterine bleeding as control.All samples were confirmed pathologically.Methylation specific PCR (MSP) was performed to detect the promoter methylation of DACH1 gene,and analyze its influence on the expression of DACH1 and the relationship between DACH1 promoter methylation and clinicopathological factors in EC.DACH1 protein expression was detected by western blot.Chi-square test and Pearson test were used for statistical analysis.ResultsThe rate of promoter methylation of DACH1 gene in the EC tissues was significantly higher than that in the normal endometrium issues (30％ vs.5％,P ＜ 0.05).There was an association between the expression of DACH1 and DACH1 gene promoter methylation ( r =- 0.30,P ＜ 0.01 ).There was statistical difference between the methylation of DACH1 and the pathological grade ( P ＜ 0.05 ) or histological type ( P ＜0.05).But DACH1 gene methylation was not related with the age,stage,myometrial invasion depth and lymphnode metastasis (P ＞ 0.05 ).Conclusions DACH1gene promoter methylaion could lead to a decrease or absence in the DACH1 expression in EC.The promoter methylation of DACH1 gene may induce the inhibition of DACH1 expression,which might be one of the mechanisms of DACH1 gene inactivation in human EC.
ABSTRACT
<p><b>OBJECTIVE</b>To establish a novel nude mice model which can be visualized in real time and detected in a continuous and dynamic way for the development and metastasis of adenoid cystic carcinoma.</p><p><b>METHODS</b>Human adenoid cystic carcinoma cells, ACCM cell line, were infected with retroviral vector of pLEGFP-N1 and then screened for a single colony of ACCM-GFP cells. Cell proliferation and morphological analysis were conducted for ACCM and ACCM-GFP cells. Nude mice lingual carcinoma model was set up with ACCM-GFP cells injection and real time observation with fluorescence imaging on ACCM-GFP tumors was performed subsequently. Histological assay was analyzed for ACCM and ACCM-GFP tumors as well.</p><p><b>RESULTS</b>ACCM-GFP cells were able to express GFP stably in the long term. ACCM and ACCM-GFP cells showed no significant difference in cell proliferation and morphology, and no significant difference of histological characteristics in vivo could be found between ACCM and ACCM-GFP tumors. Tumor development could be monitored in real time with fluorescence imaging system in vivo.</p><p><b>CONCLUSION</b>GFP-expressing ACCM tumor model can be applied to detect and observe its development in the long term in a noninvasive, real time and dynamic way. It is also a kind of ideal in vivo mouse model for adenoid cystic carcinoma research.</p>
Subject(s)
Animals , Humans , Mice , Carcinoma, Adenoid Cystic , Cell Line, Tumor , Cell Proliferation , Disease Models, Animal , Green Fluorescent Proteins , Mice, Inbred BALB C , Mice, NudeABSTRACT
Objective To investigate the effects of combination of propofol and whole-body hypoxic preconditioning on lung ischemia-reperfusion(I/R)injury in rats and the mechansim involved.Methods Ninety male SD rats weighing 250-320 g,were randomly divided into 5 groups(n=18 each): sham operation group(group S),lung I/R group(group I/R),propofol preconditioning group(group P),whole-body hypoxic preconditioning group(group WBHP),and combination of propofol and whole-body hypoxic preconditioning group(group PW).The animals were anesthetized with intraperitoneal 3% pentobarbital 30 mg/kg,tracheostomized and mechanically ventilated.Lung I/R injury was produced by occlusion of hilum of the left lung for 45 min followed by reperfusion.Propofol was continuously infused iv at 30 mg·kg-1·h-1 30 min before ischemia in group P.In group WBHP,5 times of WBHP were performed before ischemia.In group PW,propofol was infused iv at 30 mg· kg-1·h- 1 and 5 times of WBHP were performed 30 min before ischemia.Six rats from each group were killed at 30 min,1 h,and 4 h of reperfusion(T1-3).The lungs were then removed for determination of the contents of TNF-α,IL-1,IL6 and MDA,and activities of SOD.The W/D lung weight ratio was calculated.Results Compared with group S,the contents of TNF-α,IL-1,IL-6 and MDA and W/D ratio were significantly increased at T1-3,and SOD activity was significantly decreased at T1-3 in the other four groups(P＜0.05).The contents of TNF-α,IL-1,IL-6 and MDA and W/D ratio were significantly lower at T1-3 ,and SOD activity was significantly higher at T1-3 in group P,WBHP and PW than in group I/R(P ＜ 0.05).The contents of TNF-α and IL-6 and W/D ratio at T2,3 and contents of IL-1 and MDA at T3 were significantly lower,and SOD activity was significantly higher at T2,3 in group PW than in group P and WBHP(P＜0.05).There was no significant difference in the parameters metioned above between group P and WBHP(P＞0.05).Conclusion The combination of propofol and WBHP can protect the lungs from I/R injury,the efficacy is better than that of either of them alone,and it may be related to the enhancement in the inhibiton of inflammatory reaction and improvement in the antioxidant effect.
ABSTRACT
Objective To investigate the value of fluorescin-aided confocal laser endomicroscopy (CLE) for diagnosis of helicobacter pylori (Hp) infection. Methods From June 2009 to November 2009, patients undergone gastric endoscope examination with upper gastrointestinal symptoms (upper abdominal discomfort, abdominal distension, satiation, acid reflux and eructation) or screened for gastric cancer were enrolled. The gastric mucosa CLE image data of twenty diagnosed Hp positive patients and 10 Hp negative patients was analyzed retrospectively. By comparing with histological image of targeted biopsy tissue, the CLE diagnostic criteria for Hp infection were established. In the prospective study, CLE diagnose result was compared with Hp tested result. The consistency of CLE diagnostic criteria in different observers was also analyzed. The CLE image data with histopathology result were compared accordingly. Results Total 72 patients were enrolled in the prospective study,of 34 Hp positive patients, 31 patients were correctly diagnosed by CLE. The accuracy, sensitivity and specificity of CLE diagnosis were 88.9%, 91.2 and 86.8% respectively. CLE image displaying fluorescin leakage and cell shedding was the highest specificity for Hp infection diagnosis, (97.4 %);fluorescin leakage plus gastric pits distortion and cell edema was the highest sensitivity (88. 2%). The consistency of CLE diagnostic criteria in different observers was high (Kappa value 0. 72, 0.87). The CLE image of Hp infection was highly correlated with inflammation activity (P＜0. 001). Conclusion CLE can accurately distinguish normal mucosa from Hp infected mucosa at the cellular level. The diagnostic value for Hp infection was reliable.
ABSTRACT
Objective To detect the expression of a novo tumor suppressor gene PTEN and DNA direct repair enzyme MGMT in gynecomastia. Methods Immunohistochemical SP method was used to detect expression of PTEN and MGMT protein in 68 cases of gynecomastia(experiment group) and 24 cases of mammary gland of control group. The selected examples were divided into three different age groups and three different histological types. Results The PTEN and MGMT protein were all expressed in nucleusr of ductal cellula epithelialis. The expression level of PTEN and MGMT proteins in gynecomastia was significantly lower than that of mammary gland of control(P
ABSTRACT
Objective To investigate the relationship of c erbB2 with endocrine therapy and prognosis, to investigate the rapid and effective method to detect c erbB2 alteration in breast carcinoma(BC).Metheds Semi quantitative PCR was used to detect the amplification of c erbB2, and immunohistochemistry was used to detect the expression of protein of c erbB2. 58 cases of BC were followed up .Results There was significant relationship between c erbB2 protein overexpression and gene amplification(P