AIM:To study the effects of extracellular potassium on the protein expression of wild-type HERG and its mutant L539fs/47.METHODS:Wild-type HERG (WT) or its mutant HERG-L539fs/47 (MT) were transfected into HEK293 cells for 36 h.The cells were incubated in different media containing 0.8, 4.3 or 10 mmol/L potassium.Af-ter 6 h of incubation, the protein expression of HERG was detected by flow cytometry.After 12 h of incubation, the locali-zation and quantity of the proteins were detected by laser confocal imaging and Western blot.RESULTS: Different from the retention of mutant protein in cytoplasm, wild-type HERG protein was mainly distributed in the cell membrane.The 2 proteins both increased with the changes of extracellular potassium.Flow cytometry showed that the fluorescence in the 2 groups both increased with the changes of extracellular potassium ( P<0.01 ) .The fluorescence in WT group was signifi-cantly higher than that in MT group (P<0.01).Western blot showed that mutant HERG protein included only one 60 kD band, different from the 135 kD and 155 kD bands in wild-type HERG, which were affected by the changes of extracellular potassium (P<0.05).CONCLUSION:The retention of HERG mutant L539fs/47 protein in the cytoplasm is more than wild-type HERG.Chronic high extracellular potassium keeps the stability of wild-type and mutant HERG proteins on the cell membrane.Chronic low potassium reduces the expression of HERG channel proteins in a time-dependent manner.

Objective To observe the effect of somatostatin combined with transcatheter arterial infusion chemotherapy in the treatment of malignant intestinal obstruction.Methods 60 patients with malignant intestinal obstruction were enrolled in our hospital from January 2013 to June 2015.The rats were randomly divided into 4 groups: the conventional group(n=15),the growth inhibition group(n=15),the low dose intra-arterial infusion chemotherapy group(n=15)and somatostatin plus low-dose intra-arterial infusion chemotherapy group(n=15).The clinical symptom remission time,duration of clinical symptom remission and survival time were compared between the four groups.Results The study showed that somatostatin group of gastrointestinal decompression and symptom remission time were significantly better than the conventional group.Especially the symptom remission time is significantly faster than the conventional group and chemotherapy group(P< 0.05); the duration of chemotherapy group and somatostatin+artery perfusion chemotherapy group in survival time and relieve symptoms,were significantly better than the conventional group and somatostatin group(P< 0.05).Conclusion Somatostatin is beneficial to reduce the amount of gastrointestinal decompression in the treatment of malignant intestinal obstruction.Celiac artery infusion chemotherapy can help to improve the maintenance time and prolong the survival time of patients with malignant obstruction.The combination of somatostatin and celiac artery infusion chemotherapy is beneficial to relieve the symptoms and prolong the time interval,and improve the survival time of patients.

Objective To investigate the short term effect of licartin transarterial infusion in combination with chemoembolization (LTACE) and compare its effect with conventional transcatheter arterial chemoembolization (TACE) for advanced hepatocellular carcinoma (HCC).Methods Seventy-two cases of advanced HCC were included in this analysis.There were 50 males and 22 females with the average age of (58±12) yrs (range 34-86 yrs).Twenty-nine patients received LTACE treatment while the other forty three patients received conventional TACE treatment.Before intervention,there was no variation (P>0.05) in gender (X2 =0.202),Child-Pugh grading for hepatic function (X2=2.428),as well as in white blood cell count (t=1.101)and platelet count (t =0.080) between the two groups except for age and portal vein thrombosis.For LTACE group,30 minutes after the infusion of licartin (27.75 MBq/kg) into proper hepatic artery,an emulsion of 40 rag pharmorubicin and 30 ml uhrafluid lipidol was infused until hemostasis within target artery.For TACE group,only an emulsion of 40 nag pharmorubicin and 30ml uhrafluid lipidol was infused until hemostasis within target artery.Following these interventions,the two groups were given the same treatment to stabilize hepatic function and relief embolization-relating symptoms; Patients' follow-up included clinical symptoms and signs,hepatic and renal function,peripheral blood test,CT and radionuclide study(ECT).All data were analyzed with SPSS 11.5.Measurement data were expressed with mean and processed by t test; numeration data were processed by Chi square test and Fisher precise test; Kaplan-Meier analysis and log-rank test were applied for comparing the survival rate of the two groups.P <0.05 means the exist of a statistic variation.Results After treatment,there was no variation of Albumin,GPT,serum bilirubin,white blood cell,platelet and serum creatinine level between the two groups [t=0.250,0.907,0.629,0.005,0.250,0.453 (7 days) and 0.978,1.250,1.942,0.733,0.315,1.243 (14 days); P >0.05].ECT imaging demonstrated a 55.17% (16/29) uptake ratio of licartin within tumor areas by the time of 7-days follow-up study.The lesions in both LTACE and TACE groups exhibited a decrease in their size and statistically significant difference was demonstrated before and after treatment in either group( t=7.207,8.006,P <0.01).But between the two groups,the tumor size reduction showed no statistical difference,the tumor size in LTACE and TACE groups were( 1.68±0.32),(1.74±0.31)respectively (t =0.786,P>0.05)before treatment and(1.52±0.38),(1.61±0.36) respectively(t=0.891,P>0.05) after treatment.There was no variation between the two groups comparing the 6 months cumulative survival rate(LTACE 52%,TACE 76%,log-rank test,X2=3.080,P >0.05).Conclusion There was no statistically significant differences between LTACE and TACE groups concerning the short term effect and adverse reaction for treatment of advanced HCC.The long term outcomes should be established on the basis of a large-sample,multiconter,randomized trail.

Objective:To evaluate the effects of exosomes derived from cardiac fibroblasts treated with hypothermic hypoxia-reoxygenation on ventricular electrical conduction during hypothermic cardiac ischemia-reperfusion (I/R) in rats.Methods:SPF neonatal Sprague-Dawley rats of either sex, aged 1-2 days, were used, and primary cardiac fibroblasts were extracted by differential adhesion method. The cells were passaged for 2-4 generations. When the cell density reached 60%-70%, the cells were transferred and exposed to 95% N 2 + 5% CO 2 for 1 h at 4 ℃, and then exposed to 95% air + 5% CO 2 for 24-48 h at 37 ℃, and then exosomes were extracted. Twenty-four SPF healthy adult male Sprague-Dawley rats, aged 2-3 months, weighing 280-360 g, were divided into 3 groups ( n=8 each) according to the random number table method: control group (group C), hypothermic cardiac IR group (I/R group) and exosome + hypothermic cardiac IR group (Exo-IR group). At 48 h before equilibrium perfusion, 1.5 ml (200 μg) of exosomes secreted by cardiac fibroblasts treated with hypothermic hypoxia-reoxygenation was injected into the tail vein in Exo-IR group, and PBS 1.5 ml was injected into the tail vein in C group and IR group each. Group C received 110 min equilibration perfusion. After 20 min of equilibration, the perfusion was suspended for 60 min (global ischemia) followed by 30 min of reperfusion in IR and Exo-IR groups. Microelectrode arrays were applied at 20 min of equilibrium perfusion and 15 and 30 min of reperfusion to obtain myocardial conduction velocity (CV), absolute conduction inhomogeneity (P 5-95) and inhomogeneity index (P 5-95/P 50) on the left ventricular surface of isolated rat hearts. Results:Compared with group C, the CV was significantly decreased at 15 and 30 min of reperfusion, and P 5-95 and P 5-95/P 50 were increased in IR and Exo-IR groups ( P<0.05). Compared with IR group, CV was significantly increased at 15 and 30 min of reperfusion, and P 5-95 and P 5-95/P 50 were decreased in Exo-IR group ( P<0.05). Conclusions:Exosomes derived from cardiac fibroblasts treated with hypothermic hypoxia-reoxygenation can improve ventricular electrical conduction during hypothermic cardiac I/R in rats.

Background::The pharmacokinetic and clinical behaviors of many proton pump inhibitors (PPIs) in peptic ulcer treatment are altered by CYP2C19 genetic polymorphisms. This non-inferiority study evaluated the efficacy and safety of the novel PPI anaprazole compared with rabeprazole. We also explored the influence of Helicobacter pylori ( H. pylori) infection status and CYP2C19 polymorphism on anaprazole. Methods::In this multicenter, randomized, double-blind, double-dummy, positive-drug parallel-controlled, phase III study, Chinese patients with duodenal ulcers were randomized 1:1 to receive rabeprazole 10 mg + anaprazole placebo or rabeprazole placebo + anaprazole 20 mg once daily for 4 weeks. The primary efficacy endpoint was the 4-week ulcer healing rate assessed by blinded independent review. Secondary endpoints were the proportion of patients with improved overall and individual duodenal ulcer symptoms at 4 weeks. Furthermore, exploratory subgroup analysis of the primary endpoint by H. pylori status and CYP2C19 polymorphism was conducted. Adverse events were monitored for safety. Non-inferiority analysis was conducted for the primary endpoint. Results::The study enrolled 448 patients (anaprazole, n = 225; rabeprazole, n = 223). The 4-week healing rates were 90.9% and 93.7% for anaprazole and rabeprazole, respectively (difference, -2.8% [95% confidence interval, -7.7%, 2.2%]), demonstrating non-inferiority of anaprazole to rabeprazole. Overall duodenal ulcer symptoms improved in 90.9% and 92.5% of patients, respectively. Improvement rates of individual symptoms were similar between the groups. Healing rates did not significantly differ by H. pylori status or CYP2C19 genotype for either treatment group. The incidence of treatment-emergent adverse events was similar for anaprazole (72/220, 32.7%) and rabeprazole (84/219, 38.4%). Conclusions::The efficacy of anaprazole is non-inferior to that of rabeprazole in Chinese patients with duodenal ulcers.Registration::ClinicalTrials.gov, NCT04215653.

Objective:To evaluate the effects of exosomes derived from cardiac fibroblasts pretreated with sevoflurane on ventricular electrical conduction in isolated rat hearts subjected to hypothermic ischemia-reperfusion (I/R) using the multi-electrode array mapping technique.Methods:Primary cardiac fibroblasts were extracted by differential adhesion in SPF Sprague-Dawley rats of either sex.Cardiac fibroblasts of passage 2-4 were treated with 2.5% sevoflurane for 1 h, and then cultured for 24-48 h to extract exosome.SPF healthy male Sprague-Dawley rats, aged 2-3 months, weighing 280-320 g, were divided into 3 groups ( n=8 each) using a random number table method: control group (group C), I/R group and sevoflurane-pretreated cardiac fibroblast-derived exosome+ IR group (group S+ IR). Hearts were perfused for 110-min equilibration in group C. After 20 min of equilibration, the perfusion was suspended for 60 min (global ischemia) followed by 30 min of reperfusion in IR and S+ IR groups.Exosomes 1 ml (200 μg) derived from cardiac fibroblasts pretreated with sevoflurane were injected through the tail vein at 48 h before surgery in group S+ IR, and the equal volume of normal saline was injected instead in C and IR groups.The cardiac conduction velocity (CV), conduction absolute inhomogeneity (P 5-95) and inhomogeneity index (P 5-95/P 50) were obtained at 20 min of equilibration (T 0) and 15 and 30 min of reperfusion (T 1, 2) using the microelectrode array attaching to the left ventricular surface of the isolated heart. Results:Compared with group C, CV was significantly decreased and P 5-95 and P 5-95/P 50 were increased at T 1 ( P<0.05), and no significant change was found at T 2 in group S+ IR ( P>0.05), and CV was significantly decreased and P 5-95 and P 5-95/P 50 were increased at T 1, 2 in group IR ( P<0.05). Compared with group IR, CV was significantly increased and P 5-95 and P 5-95/P 50 were decreased at T 1, 2 in group S+ IR ( P<0.05). Conclusions:Exosomes derived from cardiac fibroblasts pretreated with sevoflurane can improve ventricular electrical conduction in isolated rat hearts subjected to hypothermic I/R.

Objective To develop a diagnostic model based on deep neural network for intelligent discrimination of thyroid function. Methods A total of 1616 patients ( 283 males, 1333 females, average age:52 years) who underwent thyroid imaging between May 2016 and June 2018 were selected. According to the clinical diagnosis, the 1616 cases included 299 normal thyroid cases, 876 hyperthyroidism cases and 441 hypothyroidism cases. Feature extraction and learning training were performed on 1000 training set sam-ples by two deep neural network models ( AlexNet;deep convolution generative adversarial networks ( DCGAN) ) using deep learning algorithm. Performance verifications were implemented on 616 test set samples. The con-sistency between the verification results of the two models and the clinical diagnosis was analyzed by Kappa test. Meanwhile, the time advantage of the intelligent diagnosis models was analyzed. Results The average diagnostic time of AlexNet model was 1 s/case, and the classification accuracy for normal thyroid, hyperthy-roidism, hypothyroidism were 82.29％(79/96), 94.62％(369/390), 100％(130/130), respectively. The Kappa value between results of AlexNet model and clinical diagnosis was 0.886 ( P<0.05) . The average di-agnostic time of DCGAN model was 1 s/case, and the classification accuracy for normal thyroid, hyperthy-roidism, hypothyroidism were 85.42％(82/96), 95.64％(373/390), 99.23％(129/130), respectively. The Kappa value between results of DCGAN model and clinical diagnosis was 0.904 ( P<0.05) . Conclusion The deep neural network intelligent diagnosis model can quickly determine the functional status of thyroid gland in thyroid imaging, and it has a high recognition accuracy, thus providing a new method for thyroid image review.

Objective To explore the signs of consistent changes of intestinal flora in type 2 diabetes mellitus (T2DM) and diabetes kidney disease (DKD) patients, by studying the key change characteristics of intestinal flora in these patients. Methods Thirty patients with T2DM,twenty-five patients with DKD were involved. Thirty healthy patients with matching age and sex were also involved as the control group. Fecal and serum specimens were collected from both the study group and the control group. High-throughput sequencing technology was used to sequence the 16S rDNA-v4 region of fecal samples;interleukin-6 (IL-6) and C-reactive protein (CRP) were detected by electrochemical luminescence and immunoturbidimetry. Microbiome analysis software QIIME (v1.9.1) was used to analyze the composition and diversity of intestinal flora. Microbial diversity analysis software LEfSe was used to compare intestinal bacteria markers differences between the study group and the healthy control group. The diagnosis model was established by the random forest method. The change characteristics of intestinal flora function were predicted by the PICRUSt. Results The intestinal flora diversity of DM and DKD patients was significantly different from that of the healthy control group (P<0.05). T2DM and DKD patients harbored lots of similar changes. For example, there was a significant decrease in Lachnospira, Faecalibacterium, Roseburia and Coprococcus(P<0.05). However, there was also a disease-specific pattern of imbalance between the two disease. There was a significant increase in Bacteroides in T2DM patients, and in Lactobacillus, Slackia, Anaerotruncus,Haemophilus and Enterococcus in DKD patients. Functional prediction was also confirmed that T2DM and DKD patients had more consistent changes. The correlation analysis between serum inflammatory indicators of T2DM and DKD and bacteria suggested that the decrease of beneficial bacteria in the intestinal tract of T2DM and DKD patients may be the cause of the increase of serum inflammatory indicators. Conclusion T2DM and DKD patients harbored lots of similar changes in intestinal flora, a decrease of bacteria producing butyrate,but there was also a disease-specific change between the two disease,providing a data basis for further studies to evaluate the risk of nephropathy in patients with diabetes by intestinal flora .

Background::There is a need for effective and safe therapies for psoriasis that provide sustained benefits. The aim of this study was to assess the efficacy and safety of tildrakizumab, an anti-interleukin-23p19 monoclonal antibody, for treating moderate-to-severe plaque psoriasis in Chinese patients.Methods::In this multi-center, double-blind, phase III trial, patients with moderate-to-severe plaque psoriasis were enrolled and randomly assigned (1:1) to receive subcutaneous tildrakizumab 100 mg or placebo at weeks 0 and 4. Patients initially assigned to placebo were switched to receive tildrakizumab at weeks 12, 16, and every 12 weeks thereafter. Patients in the tildrakizumab group continued with tildrakizumab at week 16, and every 12 weeks until week 52. The primary endpoint was the Psoriasis Area and Severity Index (PASI 75) response rate at week 12.Results::At week 12, tildrakizumab demonstrated significantly higher PASI 75 response rates (66.4% [73/110] vs. 12.7% [14/110]; difference, 51.4% [95% confidence interval (CI), 40.72, 62.13]; P <0.001) and Physician’s Global Assessment (60.9% [67/110] vs. 10.0% [11/110]; difference, 49.1% [95% CI, 38.64, 59.62]; P <0.001) compared to placebo. PASI 75 response continued to improve over time in both tildrakizumab and placebo-switching to tildrakizumab groups, reaching maximal efficacy after 28 weeks (86.8% [92/106] vs. 82.4% [89/108]) and maintained up to 52 weeks (91.3% [95/104] vs. 87.4% [90/103]). Most treatment-emergent adverse events were mild and not related to tildrakizumab. Conclusion::Tildrakizumab demonstrated durable efficacy through week 52 and was well tolerated in Chinese patients with moderate-to-severe plaque psoriasis.Trial registration::ClinicalTrials.gov, NCT05108766.

【Objective】 To explore the effect of Genistein on the proliferation and cell cycle regulation of ovarian cancer cells. 【Methods】 Ovarian cancer OVCAR-5 cells were treated with Genistein. Cell counting and MTS assays were performed to determine the alterations of cell proliferation. Real-time PCR and Western blotting were conducted to examine the expression changes of key cell cycle regulators. 【Results】 Genistein significantly promoted the proliferation and viability of OVCAR-5 cells. After Genistein treatment, cellular mRNA and protein expression levels of cell cycle activators such as PCNA, Cyclin D1 and CDK4 were increased, but those of cell cycle inhibitors such as p21 and p27 were decreased. 【Conclusion】 Genistein can upregulate the proliferation and G1-S transition of ovarian cancer OVCAR-5 cells. The discrepancy may be caused by diverged experimental conditions and/or different ER expression patterns of cell lines. The findings may provide basic information for in-depth analysis of the role(s) and mechanisms by which genistein confers its effect on ovarian cancer cells.