Objective To explore the expression of caspase-3 in skeletal muscle of the mice in the state of cancer, and to elucidate the relationship between Caspase-3 and apoptosis,consumption of skeletal muscle protein in cancer cachexia.Methods 48 male BALB/c mice were randomly divided into cancer cachexia group and control group (n=24).The mice in cancer cachexia group were inoculated with mouse colon 26 adenocarcinoma cells.The body weights of the mice in two groups were detected daily.Eight mice in each group were executed to test the weight of left gastrocnemius, fiber crosscut area, the expression levels of tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6),Caspase-3 proteins and the apoptotic rate of gastrocnemius cells on day 8,14,and 20,respectively. Results The mice in cancer cachexia group appeared cachectic symptoms on day 14,the non-tumor body weight was decreased more than 20% of that in control group (P<0.05).Compared with control group at the same time, the mouse body weight,non-tumor body weight,the weight of left gastrocnemius and the fiber crosscut area of the mice in cancer cachexia group were obviously decreased with the prolongation of inoculation time (P<0.05 ), whereas the expression levels of TNF-α,IL-6,Caspase-3 proteins and the apoptotic rate of muscle cells were obviously increased after tumor inoculation (P<0.05).The level of Caspase-3 protein was negatively correlated with the weight of gastrocnemius and fiber crosscut area (r=-0.716,P<0.05;r=-0.694,P<0.05),and the level of Caspase-3 was positively correlated with the levels of TNF-αand IL-6 (r=0.742,P<0.05;r=0.675,P<0.05).Conclusion Caspase-3 may be a key factor in the protein comsumption of skeletal muscle in cancer cachexia.

Objective To investigate the association between tumor necrosis factor ? (TNF ?) gene polymorphism and ankylosing spondylitis (AS).Methods Genomic DNA from 98 Chinese AS patients and 70 ethnically matched controls were typed for TNF(308) polymorphism by allele specific polymerase chain reaction (AS PCR).Results The TNF genotypes in AS patients were respectively TNF1 homozygote 37%,TNF2 homozygote 10% and TNF1 and TNF2 heterozygote 53%.While TNF genotypes in controls group were respectively TNF1 homozygote 67%,TNF2 homozygote 3% and TNF1 and TNF2 heterozygote 30%.Significant difference was found in the distribution of TNF 308 genotype between both groups ( ? 2=15 73, P

Objective To investigate the relationship between serum HBeAg level and inflammation grade (G)/fibrosis stage (S) in the liver tissues of chronic hepatitis B (CHB) patients in the immune clearance phase (IC). Methods Both liver biopsy samples and serum samples were consecutively collected from CHB patients in Liver Center,First Affiliated Hospital,Fujian Medical University during March 2007 to June 2010.Electro-chemiluminescence and fluorogenic quantitative polymerase chain reaction (PCR) methods were used to determine HBeAg titer and hepatitis B virus (HBV) DNA level,respectively.The relationships between HBeAg titer and liver pathological stages were analyzed using Spearman rank correlation analysis.Receive operating characteristic (ROC) curve was used to evaluate the diagnostic value of HBeAg for liver pathological stages.Results Totally 249 patients with CHB were enrolled into this study.The serum HBeAg absorbances in patients with liver inflammation G1 to G4 were (2.93±2.85),(2.96±2.74),(2.69±2.67) and (2.30±2.41) lg s/co,respectively,while those in patients with liver fibrosis S1 to S4 were (2.99±2.74),(2.89±2.73),(2.58±2.55) and (2.32±2.44) lg s/co,respectively,which indicated that serum HBeAg titers were significant different in patients with different grading and staging of liver tissues (x2 =47.13,P＜0.01; x2 =74.12,P＜0.01).Spearman rank correlation analysis showed that serum HBeAg titer was negatively correlated with inflammation grades and fibrosis stages of liver tissues (r=-0.418 and-0.532,respectively; both P＜0.01).ROC curve analysis revealed that the areas under the curve (AUC) were 0.74 (G≥≥3) and 0.73 (G≥4),and the HBeAg (s/co) cut-off values were 2.95 and 2.64 lg s/co,respectively.Similarly,ROC curve analysis revealed that the AUC were 0.80 (S≥3) and 0.77 S≥4),and the HBeAg cut-off values were 2.99 and 2.82 lg s/co,respectively.Conclusions The serum HBeAg titer is negatively correlated with the inflammation grades and fibrosis stages m liver tissues of CHB patients in IC phase.The level of HBeAg may be used as an adjunctive noninvasive marker to reflect the inflammation and fibrosis status in the liver.

Objective To explore the influence of tranSCription factor FOXC2 in angiogenesis of breast cancer MCF-7 cells and to clarify the action mechanism of FOXC2 in promoting tumor angiogenesis.Methods FOXC2 gene and empty vector gene were transfected into breast cancer of MCF-7 cell line with FOXC2 lentivirus gene transfection technique to obtain stable transfection cell line. The MCF-7 cells were devided into non-transfected group,empty-vector group and over-expression group.Matrigel assay and Transwell chamber test were used to observe the changes of tube formation and migration ability of human umbilical vein endothelial cells (HUVECs)in MCF-7 cells supernatant in various groups. PT-PCR and Western blotting methods were applied to detect the expressions of FOXC2,DLL4 and Notch1 mRNA and protein.Results Compared with non-tranfected group and empty-vector group,the tube formation and the migration number of HUVECs in FOXC2 over-expression group were increased(P<0.05);the expressions of FOXC2,DLL4 and Notch1 mRNA and proteins were significantly increased(P<0.05).Conclusion The FOXC2 over-expression in MCF-7 cells can increase the tube formation ability and migration ability of HUVECs,and its mechanism may be related to Notch signaling pathway.

Objective: To observe the effects of follistatin (FST)on the skeletal muscle wasting of cancer cachexia mice and the expressions of Mstn, LncRNA-MALAT1 and Caspase-3, and to elucidate its associated molecular mechanisms.Methods:Thirty-two BALB/c mices were randomly assigned into:healthy control (HC) group,FST prevention (FP)group,FST treatment (FT)group and cancer cachexia (CC)group.The murine colon adenocarcinoma CT26 cells were inoculated subcutaneously into the mices in FP, FT and CC groups to establish the cancer cachexia models. The body weight, spontaneous activity and tumor growth were daily monitored.The mice in FP and FT groups were administrated with FST intraperitoneally on day 6 and 12 after inoculation.The samples were collected on day 20.The tumor and gastrocnemius weights of the mice were detected. The biochemical metabolism indexes and myofiber cross-sectional area of gastrocnemius tissue were detected.The mRNA expression levels of Mstn,Caspase-3 and LncRNA-MALAT1 were examined by Real-time PCR.The protein expression levels of Mstn and Caspase-3 were measured by Western blotting method. Results:Compared with CC group,the body weights,spontaneous activities,gastrocnemius weights and myofiber cross-sectional areas were increased (P <0.05);the serum levels of glucose,total protein and albumin of the mice in FP and FT groups were increased (P <0.05).The protein and mRNA expression levels of Mstn and Caspase-3 in gastrocnemius of the mice in CC group were significantly higher and the expression level of LncRNA-MALAT1 was significantly lower than those in HC group (P < 0.05).The mRNA and protein expression levels of Mstn and Caspase-3 in FP and FT groups were reduced and the expression level of LncRNA-MALAT1 was increased compared with CC group (P < 0.05).The prevention effect in FP group is better than FT group (P < 0.05). Conclusion:FST may alleviate the muscle wasting of the mice with cancer cachexia by inhibiting the expression of Mstn,thus upregulating the expression of LncRNA-MALAT1 which in turn to suppress the expression of Caspase-3.

Objective The aim of this prospective observational study was to analyze the prevalence and the predictive factors of hemorrhagic events after abdominal paracentesis in patients with acute-on-chronic liver failure (ACLF).Methods ACLF patients who received at least one episode of abdominal paracentesis were prospectively enrolled between January 2010 to December 2013. Prevalences of intraperitoneal and abdomen hemorrhage complications were examined. t test was performed for continuous variables and chi-square test was performed for categorical variables.Binary Logistic regression was used to analyze the risk factors of hemorrhage.Results A total of 525 abdominal paracenteses were carried out on 185 ACLF patients within a 4-year period,with 289 (55 .0%)for diagnostic purpose and 236 (45 .0%)for therapeutic purpose.A total of 16 (3.0%)hemorrhagic complications were identified, with 4 cases of abdominal wall hematomas and 12 cases of intraperitoneal hemorrhage.Patients were divided into hemorrhage group and non-hemorrhage group according to this complication.Age,gender, Child-Pugh score,volume of ascitic fluid removed,underlying cirrhosis,platelet count and thrombin time were not significantly different between two groups (all P > 0.05 ).Patients with bleeding events had lower fibrinogen levels and higher prothrombin time,international normalized ratio,activated partial thromboplastin time and model for end-stage liver disease score (all P <0.05).After adjustment of other factors,multivariate regression analysis indicated that low fibrinogen level was the only independent predictor of hemorrhagic complication (OR=0.105,95%CI :0.018-0.608,P =0.012).Conclusion Low fibrinogen level is the independent predictor of severe hemorrhagic complications following paracenteses in patients with ACLF.

Objective To investigate the relationship between levels of ceruloplasmin (CP) and inflammation grade,fibrosis stages in liver of patients with chronic hepatitis B (CHB),and to establish liver fibrosis non-invasive model and evaluate its diagnostic value for liver pathological stages.Methods Both liver biopsy samples and sera were collected from 148 consecutive CHB patients in Liver Center,First Affiliated Hospital,Fujian Medical University during January 2009 to June 2011.The relationships between CP and liver pathological stages were analyzed using Spearman rank correlation analysis.Receiver operator characteristic (ROC) curve was used to evaluate the diagnostic value of CP for liver pathological stages.The diagnostic values of relevant indicators were analyzed by Logistic regression.The liver pathology-predicting model was built and the diagnostic value of the model was analyzed by ROC curve.Results The mean values of CP in 148 CHB patients with liver inflammation grades of G1 to G4 were (212.5 ± 34.9),(205.5± 32.0),(201.4 ± 37.7) and (172.8 ± 20.4) mg/L,respectively,which were significantly different by ANOVA test (F=6.309,P＜0.01).Similarly,the mean values of CP in patients with liver fibrosis stages of S1 to S4 were (217.4±32.3),(206.0±37.7),(194.2±29.8) and (179.7±30.4) mg/L,respectively,which were significantly different by ANOVA test (F =8.608,P ＜ 0.01).Spearman rank correlation analysis showed that CP was negatively correlated with liver inflammation grades (r=-0.316,P＜0.01) and fibrosis stages (r=-0.404,P＜0.01).ROC curve analysis revealed that the area under the curves (AUC) were 0.71 (S≥2),0.70 (S≥3) and 0.72 (S=4).Multiple Logistic regression analysis showed that CP,α-fetoprotein,cholesterol,platelet and age were independent predictors for liver fibrosis.ROC curve analysis revealed that AUC were 0.84 in model-1 (S≥2),0.83 in model-2 (S≥3) and 0.87 in model-3 (S=4).The accuracy rates were 71.8％,80.3％ and 79.2％,respectively.Conclusions The CP levels are negatively correlated with inflammation grades and fibrosis stages in the liver of CHB patients.CP could be an important non-invasive indicator for liver fibrosis and the model including CP can be used to predict liver fibrosis in CHB.

Objective: To investigate the efficacy of sequential therapy with telbivudine in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients with partial response after a standard course of interferon therapy. Methods: A retrospective cohort study was performed for 58 HBeAg-positive CHB patients with partial response at the end of interferon therapy (48-60 weeks) from January 2009 to December 2013. According to whether telbivudine was used sequentially or withdrawn at the end of the course of treatment, the patients were divided into telbivudine sequential therapy group and withdrawal group, and the two groups were compared with in terms of biochemical, virological, and serological response rates. The chi-square test, the t-test, and the non-parametric test were used based on data type. Results: A total of 58 patients were enrolled in this study, with 31 in the telbivudine sequential therapy group and 27 in the withdrawal group. At 12 and 24 weeks after interferon therapy ended, the telbivudine sequential therapy group had a significantly higher HBeAg clearance rate than the withdrawal group (22.6%/29.0% vs 0%/3.7%, P < 0.05). At week 48 of follow-up, the telbivudine sequential therapy group had a significantly higher combined response rate than the withdrawal group (22.6% vs 0%, P = 0.015). Among the 31 patients in the telbivudine sequential therapy group, 11 had an increase in creatine kinase during the administration of telbivudine. No patient in either group experienced serious adverse reactions during follow-up, such as muscular soreness, myositis, peripheral neuropathy, renal dysfunction, and liver function decompensation. Conclusion In HBeAg-positive CHB patients with partial response to interferon therapy, sequential therapy with telbivudine can increase serological HBeAg clearance rate and combined response rate at week 48, and it is safe in HBeAg-positive CHB patients achieving partial response at the end of interferon therapy.

OBJECTIVETo use a rat model of nonalcoholic liver disease (NAFLD) to observe effects of the peroxisome proliferator-activated receptor-a (PPAR-a) agonist fenofibrate on hepatic steatosis in nonalcoholic fatty liver and to investigate the underlying mechanism.

METHODSSixty-six Sprague-Dawley rats were given adaptive feeding for 1 week and then randomly allocated into the following three groups: unmodeled control (group C,n =18), untreated NAFLD model (group M, n =24), and fenofibrate-treated NAFLD model (group F, n =24).Group C rats were given a normal diet, while group M and group F rats were given a high-fat diet. After model establishment, the group F rats were treated with fenofibrate (10 mg/kg/d, intraperitoneal) and the group C and group M rats were given sham-treatment with cosolvent (5 mL/kg/d, intraperitoneal). At the end of treatment weeks 4, 6 and 8, one-third of rats in each group were euthanized.Liver tissues were assessed by hematoxylin-eosin (HE) staining to determine level of steatosis and inflammaion activity, and by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling to measure changes in hepatocyte apoptosis index. Changes in expression levels of the PPAR-a receptor and apoptosis factors (bcl-2, bax and caspase-3) were assessed by reverse transcription-PCR and immunohistochemistry.

RESULTSThe NAFLD modeled rats showed appropriate induction of hepatic steatosis, hepatic inflammation, and hepatocyte apoptosis. Compared to the group M rats, the group F rats showed lower expression of PPAR-and bcl-2 and higher expression of bax and caspase-3 at both the mRNA and protein level.

CONCLUSIONFenofibrate can ameliorate hepatic steatosis in an experimental rat model of NAFLD, and the mechanism may be associated with inhibition of hepatocyte apoptosis.

Animals ; Apoptosis ; Caspase 3 ; metabolism ; Diet, High-Fat ; Fenofibrate ; pharmacology ; Hepatocytes ; drug effects ; Non-alcoholic Fatty Liver Disease ; pathology ; Peroxisome Proliferator-Activated Receptors ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

Objective To explore the dynamic expressions and clinical significance of toll-like receptor 4 (TLR4)/microRNA (miRNA)-181a in the pathogenesis of hepatic fibrosis (HF) in patients with chronic hepatitis B (CHB) .Methods CHB patients underwent liver biopsy for fibrosis staging HF (S) .Real-time polymerase chain reaction (PCR) was used to detect the expressions of miRNA-181a in both serum and liver tissue and the expression of TLR4 mRNA in liver tissue .Western blot was used to detect the expression of TLR4 protein in liver tissue . The fibrosis-4 (FIB-4 ) index and aspartate aminotransferase-to-platelet ratio index (APRI ) were calculated for noninvasive evaluation of fibrosis staging .One-way ANOVA ,Mann-Whitney U test ,spearman correlation analysis and receiver operating characteristic (ROC) curve were used for statistical analysis .Results Forty CHB patients were includedin this study ,including 7 with S0 ,6 with S1 ,14 with S2 ,7 with S3 and 6 with S4 .Serum levesl of miRNA-181a (2-ΔΔCt ) in paitents with S0-4 were 1 .00 ± 0 .00 ,0 .68 ± 0 .08 ,1 .60 ± 0 .43 ,2 .32 ± 0 .40 , and 1 .81 ± 0 .22 ,respectively ,showing an overall upward trend (F=207 .242 ,P< 0 .01) and a positive correlation with the severity of HF (r= 0 .754 , P< 0 .01) .The expressions of miRNA-181a ,TLR4 mRNA and TLR4 protein in liver tissues showed an overall increasing trend from S 0 to S4 (F=207 .242 , 110 .390 and 57 .030 ,respectively ,all P<0 .01) .The expression of miRNA-181a in liver tissue showed a positive correlation with both the expression of TLR4 protein in liver tissue and the severity of HF (r=0 .673 and 0 .911 ,respectively ,both P< 0 .01) .There was no significant difference of APRI scores between the severe (S3-4) and non-severe (S0-2) HF groups (Z= -1 .401 ,P>0 .05) .The serum level of miRNA-181a was superior to FIB-4 index for evaluation of the severe HF (S3-4) ,with areas under the ROC curve (AUROC ) of 0 .887 and 0 .695 , respectively , and accuracy of 85 .0％ and 60 .0％ , respectively .Conclusions miRNA-181a may be involved in the regulation of TLR4 signaling pathway so that to affect the progression of HF in CHB patients ,which may be a potential new target for the prevention and early treatment of HF and a non-invasive serum marker for evaluation of HF .