Objective To study the effects of oncolytic adenoviruses ZD55 harboring IL-24 gene (ZD55-IL-24) on the apoptosis of human melanoma cell line A375. Methods The oncolytie adenoviruses ZD55-IL-24 were verified by PCR. Then, the viruses were propagated, purified, and titrated by HEK293 cell plaque assay. A375 cells were cultured, divided into three groups transfected with ZD55-1L-24, ZD55 fused with enhanced green fluorescent protein (ZD55-EGFP), and replication-deficient adenovirus ZD55 carrying IL-24 gene (AD-IL-24), respectively. The multiplicity of infection was 0.1, 1, 10 and 100, respectively.Subsequently, the eytotoxity of these viruses and proliferation of A375 cells were determined by crystal violet staining and methyl thiazolyl tetrazolium (MTT) assay, respectively. The expressions of EIA and IL-24 protein were detected by Western blot in A375 cells. Results PCR verified that the adenoviruses ZD55-IL-24 contained IL-24 gene without wild adenovirns contamination. Crystal violet staining revealed that ZD55-IL-24 had an obvious eytotoxic effect on A375 cells, and MTT assay indicated that ZD55-IL-24 inhibited the proliferation of A375 cells in a time-and concentration-dependent manner. As shown by Western blot analysis, ZD55-1L-24 expressed IL-24 and E1A protein in A375 cells with a high efficiency. Conclusions The oncolytic adenoviruses ZD55-IL-24 can efficiently express IL-24 gene, inhibit the proliferation of, and induce the apoptosis in A375 cells.

Objective To evaluate the antitumor effect of oncolytic adenovirus expressing human IL-18 gene on malignant melanoma implanted in nude mice.Methods BALB/c nude mice were subcutaneously inoculated with A375 cells to establish a model of malignant melanoma.When the volume of implanted tumor reached 100-150 mm3,murine models were randomly divided into three groups to receive a 3-day intratumoral injection of IL-18 gene-expressing human oncolytic adenovirus named ZD55-IL-18,IL-18 gene-expressing adenovirus named Ad-IL-18,phosphate buffer saline(PBS),respectively.The tumor size was measured at an interval of 4 days for 9 weeks.Hematoxylin and eosin (HE)staining was performed to observe the morphological changes of tumor cells.The protein expression of IL-18 and E1A.microvessel density in tumor tissue,and apoptosis of tumor xenografts were detected by immuno fluorescence assay,immunohistochemistry and in situ end labeling technique (TUNEL).respectively.Results The treatment with ZD55-IL-18 significantly inhibited the growth of tumor.Forty-four days after the treatment,the mean tumor volume was 1039.378±29.67 mm3 in ZD55-IL-18-treated mice.significantly smaller than that in Ad-IL-18 treated mice(2900.46±62.65 mm3)and PBS-treated mice(3980.24±63.78 mm3).HE staining showed that the nuclei of tumor cells were heavily stained with few nucleoli in ZD55-IL-18-treated mice.Increased positivity rate of IL-18 was noticed in ZD55-IL-18-treated mice vs.AD55-IL-18-treated mice(83.4%±3.2%vs 24.4%±2.1%.P＜0.01).Moreover,immunofluorescence assay revealed the presence of E1A protein in tumor tissue.A decrease was found in the microvessel density in ZD55-IL-18-treated mice compared with the PBS-treated mice(P＜0.01).The apoptosis rate in tumor cells from high to low was 86.28%±3.25%in ZD55-IL-18-treated mice,43.67%±3.46%in Ad-IL-18-treated mice,and 10.73%±2.48%in PBS-treated mice;there was a significant difference between the three groups(all P＜0.05).Conclusion The oncolytic adenovirus expressing human IL-18 gene,ZD55-IL-18,has a significant inhibitory effect on the growth and metastasis of malignant melanoma implanted in nude mice.

Objective:To identify the androgen-responsive genes in prostate and screen the molecular targets for further studying human prostate cancer. Methods:The potential androgen-responsive gene pituitary tumor transforming gene 1 (PTTG1) was selected which had been previously screened by cDNA microarray in rat prostate and its mRNA level was detected by Northern blot in the castrated rat prostate with and without replacement of Mibolerone. Immunohistochemistry was performed to determine the expression and location of PTTG1 in human prostate tissues. Then human androgen-dependent prostate cancer cells LNCaP were used as a model to study the regulation of PTTG1 by Mibolerone. Results: PTTG1 mRNA was hardly detectable in the prostate of 7-day castrated rats, while it was up-regulated dramatically in the prostate of 7-day castrated rats treated with Mibolerone for 2 days. It was showed that high expression of PTTG1 was localized to the epithelial cells of human prostate cancer but not to the stromal cells with Immunohistochemistry. Northern blot analysis indicated that LNCaP cells treated with 0.1 nmol/L Mibolerone for 2 days led to the high PTTG1 mRNA expression. The basic expression of PTTG1 in human androgen-independent prostate cancer cell lines PC3 or DU145 was even higher than that in the human androgen-dependent prostate cancer cells LNCaP treated with Mibolerone. Conclusion: Androgen can up-regulate the PTTG1 expression in castrated rat prostate and human prostate cancer cell LNCaP. It suggests that PTTG1 is potential to play an important role in human prostate cancer progression.

Objective To retrospectively study the incidence and risk factors of thyroid disease induced by peginterferon plus ribavirin (Peg IFN/RBV) antiviral treatment in patients with chronic hepatitis C (CHC).Methods From January 2012 to December 2014,a total of 362 CHC patients were treated with Peg IFN/RBV antiviral therapy at Wuxi Fifth People's Hospital,among whom 315 patients were included in this study.Thyroid function,autoantibodies and thyroid ultrasound were measured at baseline,weeks 12,24 and 48.Comparisons between groups were performed using chi-square test for qualitative data and the unpaired t test for continuous variables.Results After treatment,thyroid dysfunction developed in 12.1％ of patients.Positive anti-thyroid peroxidase and anti-thyrobolulin antibodies were detected in 21％ of patients.The thyroid ultrasound changes developed in 14.9％ of the patients,among whom normal thyroid function and thyroid autoantibodies negative were detected in 8.7 ％ of patients.The titer of thyroid autoantibodies increased steadily from baseline to the end of treatment.Logistic regression analysis showed that positive thyroid autoantibody (OR =5.423,95％ CI:3.441 8.716),sustained virological response (OR=7.201,95％CI:6.231-10.206),female(OR=1.22,95％ CI:0.951-2.212) were the risk factors of thyroid dysfunction.Conclusions Peg IFN/RBV antiviral treatment increases the incidence of thyroid disease in CHC patients.Positive thyroid autoantibody,sustained virological response and the female gender are the risk factors of thyroid dysfunction.

Objective:To analyze the relationship between antithrombin Ⅲ(AT-Ⅲ) activity and survival, bleeding and thrombosis complications in patients with acute-on-chronic liver failure (ACLF), and to explore the prediction value of AT-Ⅲ activity in the prognosis of ACLF patients.Methods:The clinical data of 130 hospitalized patients with ACLF were retrospectively collected in Wuxi No.5 People′s Hospital from January 1, 2013 to April 1, 2019. The liver function, international normalized ratio (INR), and 90-day survival rate were detected. The AT-Ⅲ activity values at admission, week two, week four, and week eight of hospitalization were recorded, and the occurrences of fecal occult blood and femoral vein thrombosis were also recorded. The measurement data were compared by t test, analysis of variance, or rank sum test, and the categorical data were compared by chi-square test. The risk factors affecting the survival of ACLF patients were analyzed by Cox regression. The survival analysis was performed using the Kaplan-Meier method. Results:At the end of 90-day follow-up of 130 patients, 56 patients died, 20 patients (15.38%) were fecal occult blood positive and 15 (11.54%) had femoral vein thrombosis. The baseline AT-Ⅲ activity in the death group was lower than that in the survival group ((17.89±13.68)% vs (36.03±11.96)%), and the difference was statistically significant ( t=-8.045, P<0.01). The baseline AT-Ⅲ activities in fecal occult blood positive and negative patients were (18.26±11.52)% and (25.06±10.97)%, respectively, and in femoral vein thrombosis and non-thrombotic patients were (17.55±10.33)% and (32.48±11.88)%, respectively. The differences were both statistically significant ( t=8.746 and 8.090, respectively, both P<0.01). Through dynamic monitoring of AT-Ⅲ, the AT-Ⅲ activity showed a downward trend in the death group, while that showed an upward trend in the survival group, but the differences were not statistically significant ( F=0.282 and 0.401, respectively, both P>0.05). The Cox regression analysis suggested INR (odds ratio ( OR)=1.364, 95% confidence interval ( CI) 1.078-1.726, P=0.010) and AT-Ⅲ activity ( OR=0.930, 95% CI 0.906-0.954, P<0.01) were the independent factors affecting the survival of patients with ACLF. The area under the receiver operator characteristic curve of the AT-Ⅲ activity for predicting 90-day survival outcome of the patient was 0.706 (95% CI 0.773-0.952, P<0.01), and the cut-off value was 25%. Patients with AT-Ⅲ activity ≥ 25% had a higher survival rate than those with AT-Ⅲ activity <25% ( χ2=58.20, P<0.01). Conclusions:AT-Ⅲ activity is associated with fecal occult blood positive and femoral vein thrombosis in ACLF patients. The AT-Ⅲ activity is an independent influencing factor for predicting the prognosis of ACLF patients. Patients with AT-Ⅲ activity less than 25% have the higher mortality rate.