Objective To investigate the influence of gene therapy using survivin as a gene target on biological behavior of pancreatic carcinoma cell line. Methods Chemically synthesized siRNA and shRNA in pGCSi vector were used to silence survivin expression of pancreatic carcinoma cell line PaTu8988. The therapeutical effects of survivin as a gene target were evaluated through determination of the down-regulation of survivin gene expression, cellular shape, cell apoptosis, cell viability and apoptosis signal pathway changes. Results After transfection of different arrays of siRNA and shRNA vectors to silence the survivin expression, survivin mRNA and protein levels were significantly decreased (P < 0.05) ; PI staining revealed the presence of karyopyknosis, the cell apoptosis index was more than 20%; hypodiploid DNA content before G0/G1 detected by flow cytometry ; cell viability measured by MTT assay was significantly decreased (P <0.05) ; the activity of caspase-3 remarkably increased (P < 0. 05). Conclusions The pancreatic carcinoma cell line PaTu8988 be induced to promote spontaneous apoptosis procedure through silencing survivin expression by RNAi, which could accelerate carcinoma cell apoptosis and improve therapeutic effect on pancreatic carcinoma.

Objective To assess the efficacy of triple therapy including proton pump inhibitor (PPI), levofloxacin and amoxicillin for the first-line treatment of H. pylori infection, and the relation between H. pylori eradication and CYP2C19 genetic polymorphism. Methods Two hundred and five H. pylori-positive patients were divided into group E_(20) (esomeprazole 20 mg twice daily), group E_(40)(esomeprazote 40 mg twice daily),group R (rabeprazole 10 mg twice daily) and group L (lansoprazole 30 mg twice daily). Besides PPI, all patients were received levofloxacin 500 mg daily and amoxicillin 1000 mg twice daily for 1 week. The CYP2C19 genotypes were detected in 161 patients. The eradication of H. pylori were analyzed by intention-to-treat (ITT) and per protocol (PP) methods.ResultsThe H. pylori eradication was 86.70% in group E_(20), 88.5% in group E_(40),73.5% in group R and 78.1% in group L. Whereas the H. pylori eradication was 90% in patients with PM genotype,81.5% in patients with HetEM genotype and 82.1% in patients with HomEM genotype. The H.pylori eradication was 83.4% and 79.00% by per protocol (PP) and intention-to-treat (ITT) analyses,respectively. There was no significant difference in H. pylori eradication among four groups (P>0.05), and no relation was found between H. pylori eradication and genotypes (P>0.05). Conclusions PPI based triple therapy was effective in eradication of H. pylori, which is not influenced by CYP2C19 genotypes.

Objective To explore the miRNA regulating the potential cancer-promoting gene CCL18 in cutaneous malignant melanoma.Methods Bioinformatics analysis was conducted by using online software miRanda and TargetScan,so as to predict the miRNA targeting CCL18 gene.Three kinds of C CL18 3'UTR dual-luciferase reporter vectors,including mutant 3'UTR vector (mutant 3'UTR group),wildtype 3'UTR vector (wild-type 3'UTR group) and empty vector (blank control group),as well as miRNA vectors carring selected miRNAs were constructed according to human gene sequence analysis,and then were used to co-transfect 293T cells.After 48-hour treatment,the cells were lysed for detection of luciferase activity.Real-time fluorescence-based quantitative PCR was performed to measure the expression of CCL 18 and selected miRNA in 14 fresh malignant melanoma tissue specimens and 14 paracancerous normal skin tissue specimens (control tissues),and their correlations were analyzed.Results Online software analysis showed that some miRNAs were identified to target the 3'UTR of CCL18 gene,including miR-183,miR-128 and miR-33a.Luciferase reporter vectors and miRNA vectors were constructed successfully.As luciferase activity assay showed,when miR-183 and miR-128 were bound to the CCL18 3'UTR,the luciferase activities were significantly higher in their mutant 3'UTR groups (11.63 ± 0.42;8.80 ± 0.49) than in their wild-type 3'UTR groups (4.86 ± 0.39;5.01 ± 0.54;both P ＜ 0.05) and blank control groups (2.41 ± 0.13;2.39 ± 0.05;both P ＜ 0.01),while there were no significant differences between miR-33a-hinding mutant 3'UTR group (6.41 ± 0.47) and miR-33a-binding wild-type 3'UTR group (6.16 ± 0.22,P ＞ 0.05).Real-time fluorescence-based quantitative PCR revealed higher mRNA expression of the CCL18 gene (3.52 ± 1.68),but lower expression of miR-183 (0.49 ± 0.32),miR-128 (0.30 ± 0.20) and miR-33a (0.46 ± 0.40) in the malignant melanoma tissues compared with the control tissues.The mRNA expression of the CCL18 gene was negatively correlated with the expression of miR-128 (rs =-0548,P ＜ 0.05),but showed no significant correlations with the expression of miR-183 and miR-33a (both P ＞ 0.05).Conclusion miR-128 may play a role in regulating the potential malignant melanoma-promoting gene CCL18.

Objective To investigate the value of endoscopic retrograde appendicitis therapy ( ERAT) in the diagnosis and treatment of atypical acute appendicitis. Methods All the 48 patients suspected of atypical acute appendicitis in Jiangsu Province Hospital from January 2015 to December 2016 were randomly divided into ERAT group and conservative treatment group according to the treatment method. The final appendectomy rate of the two groups was analyzed. Results Only 17 of the 24 patients in the ERAT group received endoscopic treatment because of complex conditions or personal wishes, and 16 cases were diagnosed as acute appendicitis. Surgical resection was performed in 5 cases because of recurrence of the disease after ERAT, and the appendectomy rate was 31. 2％ ( 5/16 ) . In the conservative treatment group, all 24 patients were treated with antibiotics. Twenty of them underwent surgical resection with appendectomy rate of 83. 3％ ( 20/24) , and 1 of them had appendiceal perforation. The appendectomy rate of the ERAT group was significantly lower than that of the conservative treatment group (χ2=11. 111, P<0. 05) . Conclusion ERAT has a high diagnostic and therapeutic value for atypical acute appendicitis.

Objective: To investigate the correlation between the diversity of biliary bacterial flora and the recurrence of common bile duct stones. Methods: From September 2018 to March 2019, 48 patients underwent endoscopic retrograde cholangiopancreatography (ERCP) at the First Affiliated Hospital of Nanjing Medical University were enrolled and divided into primary choledocholithiasis (primary group, 38 patients) and recurrent choledocholithiasis group (recurrent group, 10 patients). The bile of the patients of the two groups was collected and analyzed with high-throughput sequencing of 16S rRNA. The Chao1 estimator indicated the richness of the biliary bacterial flora, and Shannon index and Simpson index demonstrated the diversity of biliary bacterial flora. The bacterial flora distribution was explored from different levels of phylum, class, order, family, genus and species. Chi-square test and t test were performed for statistical analysis. Results: The Chao1 estimator of the primary group was higher than that of the recurrent group (419.413±118.704 vs. 396.000±70.483), and the difference was statistically significant (t=3.130, P=0.003). At the level of the phylum, the highest abundance of bacteria in the primary group was Firmicute (56.48%), the highest abundance of bacteria in the recurrent group was Proteobacteria (57.79%). The dominant bacteria of the recurrent group was Proteobacteria, which was inconsistent with that of the primary group, and there were significant differences in the distribution of Proteobacteria and Firmicute between two groups (χ²=0.962 and -2.619, both P<0.05). At the genus level, the abundance of Bacillus and Lactococcus of the recurrent group were both lower than those of the primary group (9.75% vs. 20.77%, 10.86% vs. 22.01%, respectively), and the differences were statistically significant (χ²=0.354 and 0.503, both P<0.05). The abundance of Morganella of the recurrent group was higher than that of the primary group (9.00% vs. 0.44%), and the difference was statistically significant (χ²=-2.025, P=0.049). At the species level, the abundance of Bacillus unclassified, Carnobacterium maltaromaticum and Bacillus circulans of the primary group were all higher than those of the recurrent group (17.78% vs. 8.84%, 2.39% vs. 1.11%, 2.59% vs. 0.74%, respectively), and the differences were all statistically significant (χ²=2.540, 2.643 and 2.515, all P<0.05). The abundance of Aeromonas veronii of recurrent group was higher than that of the primary group (2.04% vs. 0.01%), and the difference was statistically significant (χ²=-2.397, P=0.021). The bacteria that had significant effects in the primary group included Lactococcus (P=0.012), Lactobacillus (P=0.033) and Geobacillus (P=0.021), while in the recurrent group, which included Enterobacter (P=0.007), Aeromonadaceae (P=0.001), Actinomycetes (P=0.009), and Aeromonas (P=0.001). Conclusions There are differences in abundance and composition of biliary bacterial flora between the recurrent group and the primary group. It remains to be further studied whether the changes of some bacterial flora correlated with the recurrence of common bile duct stones.

We conducted a randomized, open-label, parallel-controlled, multicenter trial on the use of Shuanghuanglian (SHL), a traditional Chinese patent medicine, in treating cases of COVID-19. A total of 176 patients received SHL by three doses (56 in low dose, 61 in middle dose, and 59 in high dose) in addition to standard care. The control group was composed of 59 patients who received standard therapy alone. Treatment with SHL was not associated with a difference from standard care in the time to disease recovery. Patients with 14-day SHL treatment had significantly higher rate in negative conversion of SARS-CoV-2 in nucleic acid swab tests than the patients from the control group (93.4% vs. 73.9%, P = 0.006). Analysis of chest computed tomography images showed that treatment with high-dose SHL significantly promoted absorption of inflammatory focus of pneumonia, which was evaluated by density reduction of inflammatory focus from baseline, at day 7 (mean difference (95% CI), -46.39 (-86.83 to -5.94) HU; P = 0.025) and day 14 (mean difference (95% CI), -74.21 (-133.35 to -15.08) HU; P = 0.014). No serious adverse events occurred in the SHL groups. This study illustrated that SHL in combination with standard care was safe and partially effective for the treatment of COVID-19.
COVID-19 ; Humans ; Medicine, Chinese Traditional ; Research ; SARS-CoV-2 ; Treatment Outcome