To investigate the expression of hypoxia-inducible factor 1α (HIF-1α) and CD133 in predicting pathologic remission and survival of patients with locally advanced rectal cancer undergoing neoadjuvant chemoradiotherapy.One hundred and fourteen patients with locally advanced rectal cancer undergoing neoadjuvant chemoradiotherapy from January 2010 to December 2015 in Jinhua Municipal Central Hospital were enrolled in the study. RT-PCR and immunohistochemistry methods were used to detect the mRNA and protein expression of HIF-1α and CD133 before and after chemoradiotherapy. Spearman rank correlation was used to analyze the correlation between HIF-1α and CD133 mRNA expression. Univariate and logistic multivariate analyses were used to determine the factors related to pathological complete response (pCR). Logistic regression analysis and Cox's proportional hazard model were used to determine factors related to overall survival and recurrence-free survival.The expression of HIF-1α and CD133 mRNA was correlated with pT, ypTNM, pCR, recurrence and metastasis of rectal cancer, while not correlated with sex, age and BMI of patients. HIF-1α mRNA expression was positively correlated with CD133 mRNA expression (=0.579,=0.000). Immunohistochemistry analysis showed that residual cancer cells strongly expressing HIF-1α also expressed CD133 strongly. Univariate analysis showed that HIF-1α mRNA and CD133 mRNA were significantly correlated with pCR (=0.001,=0.022, respectively). Multivariate analysis showed that HIF-1α and CD133 mRNA expression were independent prognostic factors of pCR (=0.012,=0.047, respectively). Cox regression analysis showed that the expression of HIF-1α mRNA and CD133 mRNA were independent predictors of recurrence-free survival and overall survival (=0.025,=0.033, respectively).The study indicates that HIF-1α and CD133 can predict pathological complete remission and survival of patients with locally advanced rectal cancer.
AC133 Antigen ; analysis ; genetics ; Biomarkers, Tumor ; analysis ; Chemoradiotherapy ; Disease-Free Survival ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; analysis ; genetics ; Male ; Neoadjuvant Therapy ; Neoplasm Grading ; Neoplasm Metastasis ; diagnosis ; Neoplasm Recurrence, Local ; epidemiology ; genetics ; Neoplasm, Residual ; genetics ; Prognosis ; Proportional Hazards Models ; Rectal Neoplasms ; chemistry ; epidemiology ; genetics ; therapy ; Survival Rate

OBJECTIVETo establish a diet-induced obesity model in zebrafish larvae.

METHODSAt 7 days post-fertilization (dpf), 200 zebrafish larvae with normal development were randomly allocated to two groups with the feeding quantity of 30 mg per day (normal feeding group) or 180 mg per day (overfed group) for 20 days. The weight, length, BMI, triglyceride (TG) and total cholesterol (TCH) of each group were measured. Whole-mount Oil Red O staining, frozen Oil Red O staining and hematoxylin-eosin (HE) staining were used to estimate the rate of hepatic steatosis and liver histology of the zebrafish. The dynamic change of hepatic lipid droplets and distribution of adipose tissue were observed with Nile Red staining in overfed zebrafish in vivo.

RESULTSThe weight, length, BMI and TG of overfed zebrafish were significantly increased compared with those in normal feeding group. Whole-mount Oil Red O staining showed that the percent of hepatic steatosis in overfed group (89.4%) was markedly higher than that in normal feeding group (20.7%). Macrovesicular steatosis was observed in the liver of the overfed larvae. Nile Red staining visualized hepatic lipid droplets and the distribution of larval adipose tissue, which increased with feeding time in the overfed zebrafish. Starving larvae showed depletion of fat and hepatic lipid, and adipose tissue was induced after refeeding.

CONCLUSIONSWe successfully established an diet-induced obesity model in zebrafish larva, in which Nile Red staining allows in vivo observation of the adipocytes and hepatic lipid droplets.

Adipose Tissue ; Animals ; Cholesterol ; Diet ; adverse effects ; Disease Models, Animal ; Fatty Liver ; Larva ; Lipids ; Obesity ; pathology ; Triglycerides ; Zebrafish

OBJECTIVETo establish a zebrafish model of liver fibrosis via diethylnitrosamine (DEN)-induced liver injury.

METHODSA total of 120 wild-type 3-month-old zebrafish were randomly divided into DEN-treated group and control group. The survival rate and behavioral changes of each group were observed. After treatment with DEN for 2, 4, and 6 weeks, liver index was measured, and liver fibrosis was evaluated with HE staining, Gomori staining and Sirius red staining.

RESULTSNo obvious behavioral change was observed in DEN-treated group during the experiment. Compared with that in control group, the liver index of zebrafish in DEN-treated group showed no significantly changes at the time points of observation. Proliferation of reticulate fibers was found in 30% of zebrafish treated with DEN for 4 weeks, and the rate increased to 80% at 6 weeks when reticulate fibers and collagen fibers actively proliferated to result in fiber collapse and formation of fibrotic nodules.

CONCLUSIONA stable zebrafish liver fibrosis model was successfully established by inducing liver damage to facilitate studies of the pathogenesis of liver fibrosis and screening therapeutic drugs.

Animals ; Diethylnitrosamine ; Disease Models, Animal ; Liver Cirrhosis ; chemically induced ; Zebrafish

Objective To investigate the value of protective transverse colostomy in decreasing postoperative anastomotic leakage and stricture after laparoscopic resection for rectal cancer.Methods The clinical data of 128 patients with rectal cancer who were admitted to the Jinhua Hospital of Zhejiang University from March 2008 to February 2012 were retrospectively analyzed.All the patients received laparoscopic anterior resection of rectal cancer after neoadjuvant chemoradiotherapy.Sixty-one patients received protective transverse colostomy (colostomy group) and 67 patients did not receive protective transverse colostomy (non-colostomy group).The incidences of postoperative anastomotic leakage and stricture,condition of the patients and the prognosis of the patients in the 2 groups were compared.Patients were followed up via out-patient examination,in-patient chemotherapy or phone call till May 2013.All data were analyzed using the t test,chi-square test or Fisher exact probability.Results The overall incidence of postoperative anastomotic leakage was 7.03％ (9/128).The incidences of postoperative anastomotic leakage in the colostomy group and non-colostomy group were 6.56％ (4/61) and 7.46％ (5/67),with no significant difference between the 2 groups (x2 =0.000,P ＞ 0.05).The systemic condition of 4 patients who were complicated with anastomotic leakage in the colostomy group was comparatively better,and all of them were cured by conservative treatment.The condition of 5 patients who were complicated with anastomotic leakage in the non-colostomy group was severe,and 4 of them received reoperation,with the reoperation rate of 4/5 ; the other patient received conservative treatment.There was significant difference in the reoperation rate between the 2 groups (P ＜0.05).The overall incidence of postoperative anastomotic stricture was 13.28％ (17/128).The incidences of postoperative anastomotic stricture in the colostomy group and the non-colostomy group were 19.67％ (12/61) and 7.46％ (5/67),with significant difference between the 2 groups (x2=4.133,P ＜ 0.05).The incidences of severe anastomotic stricture of the colostomy group and the non-colostomy group were 8.20 (5/61) and 0,respectively.Conclusion Protective transverse colostomy could not reduce the incidence of postoperative anastomotic leakage,but could mitigate the symptoms followed anastomotic leakage and reduce the reoperation rate.Protective transverse colostomy may improve the incidence of anastomotic stricture,therefore it should be applied cautiously.

Objective To evaluate the value of transient elastography (TE) for predicting severity of liver fibrosis in patients with chronic hepatitis B (CHB).Methods A total of 969 patients with CHB was enrolled and recruited for analysis,which had been received TE scan,including 258 patients of liver biopsy,and 117 patients of gastric endoscopy.Results A total of 35 patients was excluded from analysis due to TE failure or unreliable TE.Liver stiffness measurement (LSM) was independently influenced by bilirubin,AST,liver fibrosis and inflammation,ultrasonic score and albumin.TE predicted Child-Pugh C,B/C,liver fibrosis S4,≥S3 and ≥ S2 with respective area under receiver operating characteristics curves (AUROC)0.907 (95% CI 0.886-0.928 ),0.920 ( 95% CI 0.899-0.940 ),0.871 ( 95% CI 0.819-0.923 ),0.852(95%CI0.805-0.899) and 0.807(95% CI0.749-0.865),respectively.While LSM ＜32.2 kPa excluded Child-Pugh C with 99.4% probability,LSM ≥35.3 kPa determined Child-Pugh B/C with positive predictive value (PPV) 0.820.For compensated CHB,cut-offs of LSM 23.3,15.2 and 10.8 kPa diagnosed cirrhosis,liver fibrosis ≥S3 and ≥S2 with positive likelihood ratio nearly 10.0 and PPV 0.692,0.882 and 0.980,respectively; and cut-offs 8.8 kPa,6.6 kPa excluded cirrhosis,liver fibrosis ≥ S3 with negative likelihood ration nearly 0.1 and negative predictive value 0.977 and 0.903,respectively.Correlation coefficient between LSM and grades of esophageal varices was only 0.180,and AUROC for TE predicting EV was of no clinical value.ConclusionTE relatively make accurate prediction in the severity of liver fibrosis and classification of Child-Pugh.Patients with LSM ≥ 10.8 kPa should be considered for receiving antivirus treatment.

OBJECTIVETo establish a simple and practicable method to identify the different genotypes of hepatitis B virus (HBV).

METHODSBased on the alignment of 114 complete nucleotide sequence for HBV DNA of different genotypes, the specific sequence of each genotype was found. Six primer sets were designed for each of the six genotypes according to the genotype-specific sequence, and used separately for PCR. The genotype of HBV was identified according to the positive result of PCR. Three primer sets for B, C and D genotypes were added into a single tube for PCR reaction, and HBV was genotyped according to the length of the amplified DNA.

RESULTSThere was no difference in the genotyping result of PCR by single or multiplex primers, which was identical to the PCR-RFLP method.

CONCLUSIONSThis multiplex PCR method is simple, precise, sensitive, and easy to use.

DNA Primers ; Databases, Nucleic Acid ; Genotype ; Hepatitis B virus ; classification ; genetics ; Humans ; Polymerase Chain Reaction ; methods ; Sequence Alignment ; methods