Objective Based on the observation of the changes of symptoms, histopathology, visceral sensitivity, mast cell activation, autophagy, and Beclin-1 and Claudin-2 expression in rats, we established and evaluated a new rat model of diarrhea-predominant irritable bowel syndrome (D-IBS) induced by restraint-stress combined with capsaicin (CAP) administration.Methods Forty healthy 5-week old male Sprague Dawley (SD) rats were randomly divided into normal group, model group I, model group II and model group III, with 10 rats in each group.The D-IBS model was established by restraint-stress combined with intragastric administration of CAP (2 mL/100 g body weight, 0.125% in group I, 0.250% in group II, 0.500% in group III), tail clipping and forelimb restriction for 30 minutes every day for 2 weeks.The rats in the control group were treated with saline for 2 weeks.The number of contraction of abdominal wall and arched back were measured by Power Lab instrument.The mast cell activation was detected using aldehyde-magenta-orange G staining.Light and electron microscopic examinations were performed to detect the morphology and autophagy of colonic tissues.The expressions of Beclin-1 and Claudin-2 in the colonic mucosa were detected by streptavidin-biotin complex (SABC) immunohistochemical staining.Results All rats in the model group III died during the experiment.Compared with the control group and model group I, the stool frequency was increased and the visceral sensitivity threshold decreased in the model group II, and there were statistically significant differences between the model group II and the control and model groups I (P < 0.05).The colonic mucosa, mucosal epithelium and glands in each group showed normal morphology and there was no submucosal vasodilatation and diffuse inflammatory cell infiltration.Except for the control group, round purple-reddish staining spots were observed in the rat mucosal stroma or submucosa in the model groups I and II, indicating an increased expression of mast cells.The autophagy, expressions of Beclin-1 and Claudin-2 in the colonic epithelium were significantly increased in the model group II compared with control group and model group I (P< 0.05).Conclusions The model of D-IBS induced by restraint-stress combined with capsaicin is characterized by increased diarrhea, visceral hypersensitivity, increased mast cell expression and autophagy of intestinal epithelial cells, and disruption of the intestinal mucosal barrier.This model is simple to set up and shows similar symptoms of human irritable bowel syndrome.Therefore, it is worthy of popularization and application.

Objective To prepare vardenafil hydrochloride orally disintegrating tablets and evaluate their quality. Methods The tablets were prepared by direct power compression method, using crosslinking povidone ( PVPP ) as disintegrants. The preparation method was optimized by response surface test using amount of PVPP, menthol and taste-masking agents as factors with disintegrating time and distance of bitterness as index. The results of taste of orally disintegrating tablets were determined by electronic tongue, comparing to the results of taste tests. At the same time, the properties of the tablets were evaluated using appearance, content uniformity, disintegrating time, et al. as index. Results The optimal formula was as follows:PVPP 13. 26%, menthol 0. 43%, taste-masking agent SGxj 1. 26%. The results on evaluation of electronic tongue were consistent with the results of taste tests. The quality of the prepared tablets was in line with standard. The disintegrating time was (22. 34 ± 0. 34 ) s. Conclusion The preparation technology of orally disintegrating tablets is simple, and controllable in quality.

Objective To study the expressions of DD3 mRNA and PSA mRNA in the prostate tissues and its diagnostic value in prostate cancer (PCa). Methods DD3 mRNA and PSA mRNA were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction ( FQ-RT-PCR) based on Taqman technique in the tissues of 21 cases of PCa and 39 cases of BPH. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficacy of DD3 mRNA, PSA mRNA and DD3 mRNA/PSA mRNA. Results The expressions of DD3 mRNA and PSA mRNA, and DD3 mRNA/ PSA mRNA were significantly higher in PCa tissues than those in BPH tissues ( P 0.05 for all). The AUC-ROC of DD3 mRNA,PSA mRNA and DD3 mRNA/PSA mRNA were 0. 937 (95% CI,0. 879 -0. 995) , 0.755(95% CI,0.629 -0.880) and 0.839 (95%CI,0.738 -0.940),respectively. The sensitivity for DD3 mRNA,PSA mRNA and DD3 mRNA/PSA mRNA was 90. 5% ,81. 0% and 81. 0% , respectively, and the specificity was 85.0% ,62.0% and 66.7% at cutoff value of 1.4?105 copies/mg tissue,3.0?107 copies/ mg tissue and 5. 0?10-3,respectively. The sensitivity and specificity of simultaneous detection for DD3 mRNA and PSA mRNA were 100% and 85.0%. Conclusions Both DD3 mRNA and PSA mRNA expressions were significantly higher in PCa tissues than those in BPH tissues; and the quantitative detection of DD3 mRNA is more helpful for the diagnosis. The simultaneous detection of DD3 mRNA and PSA mRNA can improve the sensitivity in the diagnosis of PCa.

Objective:To investigate the effects of methyltransferase-like protein 14 (METTL14)-mediated long-chain non-coding RNA EIF3J antisense RNA1 (Inc EIF3J-AS1) on the migration and invasion of cholangiocarcinoma cells and its mechanism.Methods:From September 2017 to December 2018, 10 pairs of cholangiocarcinoma and adjacent normal tissues were collected from the First Affiliated Hospital of Naval Medical University, which were surgically resected and pathologically confirmed. The expression of METTL14 mRNA and Inc EIF3J-AS1 in cholangiocarcinoma tissues was detected by fluorescence quantitative PCR, and the protein expression of METTL14 was detected by Western blotting. Cholangiocarcinoma cell lines HUCCTI and RBE were divided into control group and METTL14 or Inc EIF3J-AS1 knockdown group. The corresponding normal lentivirus was transfected in the control group, and METTL14 or Inc EIF3J-AS1 knockdown group was transfected with lentivirus that interfered with the expression of METTL14 or Inc EIF3J-AS1, respectively. Transwell assay was used to detect the ability of cell migration and invasion, and Western blotting was used to detect the expression of epidermal growth factor receptor (EGFR) and AKT protein.Results:The expressions of METTL14 mRNA and lnc EIF3J-AS1 in cholangiocarcinoma tissues were significantly higher than those in adjacent normal tissues (0.075±0.012 vs 0.031±0.006, 0.140±0.032 vs 0.064±0.012), and there was a positive correlation between expression of METTL4 mRNA and expression of lnc EIF3J-AS1 ( r=0.883, P=0.0007). The expression of METTL14 protein in cholangiocarcinoma tissues was higher than that in adjacent normal tissues (0.354±0.131 vs 0.187±0.183). Compared with the control group, the expression of lnc EIF3J-AS1 was significantly lower in METTL14 or Inc EIF3J-AS1 knockdown group (0.217±0.020 vs 1.000±0.052, 0.149±0.066 vs 1.000±0.045). The migration and invasion ability of cell lines HUCCTI and RBE decreased significantly in lnc EIF3J-AS1 knockout group (5.00±0.58 vs 23.33±0.33, 20.33±0.67 vs 70.67±0.33; 12.00±0.58 vs 25.00±2.52, 22.33±0.89 vs 43.67±0.33). The expression of EGFR and p-AKT/AKT protein were also significantly decreased (0.109±0.015 vs 1.000±0.018, 0.226±0.036 vs 1.000±0.051; 0.118±0.052 vs 1.000±0.069, 0.132±0.098 vs 1.000±0.023). The above differences were statistically significant (all P<0.05). Conclusions:Abnormal expression of lnc EIF3J-AS1 in cholangiocarcinoma mediated by METTL14 can promote tumor cell migration and invasion.

Background and aim:Real-world data on the effectiveness and safety of treatment with the direct-acting antiviral agent-based regimen are limited on the Chinese mainland.The aim of this study was to conduct a multicenter,prospective,real-world study of ombitasvir/paritaprevir/ritonavir(OBT/PTV/r)combined with dasabuvir(DSV)in hepatitis C virus(HCV)genotype 1b-infected non-cirrhotic or compensated cirrhotic Chinese adult patients. Materials and methods:Genotype 1b-infected patients were enrolled at eight sites in China.Patients received 25/150/100 mg of OBT/PTV/r once daily combined with 250 mg of DSV twice daily for 8 weeks or 12 weeks.Sustained virological response at 12 weeks post-treatment(SVR12)and the incidence of adverse events were assessed.We have also evaluated the effect of intensive questioning of patients who were overdue for SVR12 testing.Intention-to-treat(1TT)and modified 1TT(mITT)populations were used in the analysis. Results:One hundred forty patients were included,among whom 90.0％(126/140)were newly diag-nosed,9.3％(13/140)had compensated cirrhosis,92.9％(130/140)received 12 weeks of treatment,and 7.1％(10/140)received 8 weeks of treatment.In the mITT population,the virological response rate at week 4 was 96.4％(108/112),and at the end of treatment was 100％(102/102).Among these patients,139 patients completed 12 weeks of treatment,and 73 patients were followed-up.All followed-up patients achieved SVR12.There was no adverse event-related discontinuation.Serious adverse events during treatment were reported in two(1.4％)patients,and none were considered to be drug-related.Sixty-six(47.1％)patients did not return to receive the HCV RNA test at 12 weeks post-treatment. Conclusions:The rate of SVR12 was consistent with Phase Ⅲ clinical studies.OBT/PTV/r combined with DSV showed effectiveness in Chinese adult patients,and both tolerability and safety profile were favorable.However,patient compliance should be further improved in the real world.