MiR-142-3p enhances chemosensitivity of breast cancer cells and inhibits autophagy by targeting HMGB1

MiR-142-3p has been reported to act as a tumor suppressor in breast cancer. However, the regulatory effect of miR-142-3p on drug resistance of breast cancer cells and its underlying mechanism remain unknown. Here, we found that miR-142-3p was significantly downregulated in the doxorubicin (DOX)-resistant MCF-7 cell line (MCF-7/DOX). MiR-142-3p overexpression increased DOX sensitivity and enhanced DOX-induced apoptosis in breast cancer cells. High-mobility group box 1 (HMGB1) is a direct functional target of miR-142-3p in breast cancer cells and miR-142-3p negatively regulated HMGB1 expression. Moreover, overexpression of HMGB1 dramatically reversed the promotion of apoptosis and inhibition of autophagy mediated by miR-142-3p up-regulation. In conclusion, miR-142-3p overexpression may inhibit autophagy and promote the drug sensitivity of breast cancer cells to DOX by targeting HMGB1. The miR-142-3p/HMGB1 axis might be a novel target to regulate the drug resistance of breast cancer patients.     

miR-142-5p通过影响上皮间质转化抑制肺腺癌H1650细胞的侵袭与迁移

目的：探讨肺腺癌组织中miR-142-5p的表达及其对H1650细胞增殖、侵袭、迁移及上皮间质转化（epithelieal-mesenchymal transition,EMT）的影响及其作用机制。方法：收集2014年1月至2015年1月在河北医科大学第四医院胸外科行肿瘤切 除并经病理证实的107例肺腺癌患者的癌组织及其癌旁组织标本,以及人肺腺癌细胞系H1650、HCC827、 A549、 H1975、PC9和人 支气管上皮细胞BEAS-2B, 用qPCR实验检测肺腺癌组织及细胞中miR-142-5p的表达水平及其与患者临床特征的关系。分别用 miR-142-5p模拟物（mimics）、miR-阴性对照质粒（miR-NC）转染H1650细胞后, 用CCK8、细胞划痕愈合和Transwell侵袭实验分 别检测H1650细胞的增殖、侵袭和迁移能力。使用生物信息学工具预测miR-142-5p的靶基因,通过双荧光素酶报告基因实验验 证miR-142-5p对靶基因的调控作用,Western blotting检测细胞周期蛋白依赖性激酶5（cyclin-dependent kinase 5,CDK5）及EMT 相关蛋白的表达水平。结果：肺腺癌组织及细胞系中miR-142-5p表达水平显著低于癌旁组织及BEAS-2B细胞（均P<0.01）;107 例肺腺癌组织中, 61例（57.01%）低表达miR-142-5p,其表达水平与患者的TNM分期、淋巴结转移密切相关（均P<0.01）。转染 miR-142-5p模拟物后, H1650细胞中miR-142-5p高表达,细胞的增殖、侵袭和迁移能力显著降低（均P<0.05或P<0.01）。生物信息 学工具预测CDK5是miR-142-5p的靶基因,经双荧光素酶报告基因验证,miR-142-5p可显著降低H1650细胞中CDK5的表达水 平,显著提高E-cadherin表达,降低N-cadherin和Snail的表达水平（均P<0.01）。结论：miR-142-5p在肺腺癌组织和细胞中呈低表 达状态,其通过下调CDK5表达影响EMT抑制H1650细胞的侵袭与迁移能力。     

miR-142-3p靶向调控mll-af4融合基因的实验研究

本研究探讨mll-af4融合基因的表观遗传学调控机制,以筛选靶向调控mll-af4融合基因的microRNA。利用Targetscan在线分析软件预测特异性靶向mllaf4基因3′端非翻译区域（3′-untranslated region,3′UTR）的microRNA,采用PCR方法从1例健康供者DNA中扩增mll-af4基因3′UTR序列,插入经EcoRI和PstI双酶切的荧光素酶报告载体pGL3-M,采用脂质体SuperFect包裹荧光素酶重组质粒及microRNA表达质粒转染293T细胞,应用双荧光素酶检测试剂盒测定荧光素酶活性。miR-142-3p mimics以脂质体Hiperfect转染至RS4;11细胞,选用实时定量PCR及Western blot检测mll-af4 mRNA及蛋白的表达。结果表明,成功构建了含有1935bp、2104bp和1371bp的mll-af4基因3′UTR序列的荧光素酶报告重组质粒pGL3-AF4-3′UTR,并通过酶切及基因测序方法鉴定得到证实。荧光素酶报告实验提示,miR-142组荧光素酶活性明显低于对照组。RS4;11细胞中过表达miR-142-3p可以明显下调MLL-AF4融合蛋白及mRNA表达。结论：miR-142-3p通过靶向结合mll-af4基因3′UTR的结合位点特异调控mll-af4基因表达。     

Production and characterisation of a recombinant scFv reactive with human gastrointestinal carcinomas

SC142-reactive antigen are highly glycosylated glycoproteins expressed on tissues of gastric and colon cancers but not on normal tissues. Murine SC142 antibody specific for the SC142-reactive antigen has been produced by immunisation with SNU16 stomach cancer cells. However, SC142 antibody has several potential problems such as high immunogenicity and poor tumour penetration owing to their large size. To improve tumour penetration potential in vivo, recombinant single-chain fragments have been produced using the original hybridoma cells as a source of variable heavy- and variable light-chain-encoding antibody genes. The use of the polymerase chain reaction, expression cloning technology and gene expression systems in E. coli has led to the production of SC142 single-chain fragments, which was similar in activity to the SC142 parent antibody confirmed by immunohistochemistry. Analysis by DNA sequencing, SDS-PAGE and Western blotting has demonstrated the integrity of the single-chain fragments. Competitive ELISA showed that SC142 single-chain fragments originated from parent SC142 antibody. BIAcore biosensor binding experiments showed that the SC142 single-chain fragments had an ideal dissociation rate constant as a tumour imaging reagent. These results illustrate the potential application of these novel products as an immunodiagnostic and further immunotherapeutic reagent.     

Cardioprotective functions of atrial natriuretic peptide and B-type natriuretic peptide: a brief review

1. If one was to design a hormone to protect the heart, it would have a number of features shown by the cardiac natriuretic peptides atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP). These hormones are made in cardiomyocytes and are released into the circulation in response to atrial and ventricular stretch, respectively. Atrial natriuretic peptide and BNP can reduce the preload and after-load in normal and failing hearts. They reduce blood volume over the short term by sequestering plasma and over the longer term by promoting renal salt and water excretion and by antagonizing the renin-angiotensin-aldosterone system at many levels. Each of these actions affords indirect benefit to a volume- or pressure-threatened heart. 2. Recent studies have identified additional modes of action of the natriuretic peptides that may also confer cardioprotective benefits, especially in heart disease. The emerging findings are: (i) that ANP and BNP antagonize the cardiac hypertrophic action of angiotensin II and continue working under conditions where endothelial nitric oxide (NO) function is compromised, such as in the presence of high glucose in diabetes; (ii) they potentiate the bradycardia caused by inhibitory ('autoprotective') cardio-cardiac reflexes; and, furthermore, (iii) BNP can suppress cardiac sympathetic nerve activity in humans, including those with heart failure. Thus, it appears that natriuretic peptides can shift sympathovagal balance in a beneficial direction (away from the sympathetic). The vagal reflex and antihypertrophic actions of the peptides are mediated by particulate guanylyl cyclase (pGC) natriuretic peptide receptors. 3. The multiple synergistic actions of the natriuretic peptides make them and their pGC receptors attractive targets for therapy in heart disease. Encouragingly, exogenous natriuretic peptides remain effective even when endogenous peptide levels are raised, as is the case in heart failure. They also remain effective in disease states where other protective mechanisms, such as the NO system, have become ineffective, offering yet further encouragement for the therapeutic use of the natriuretic peptides.     

Diagnosis of olfactory disturbance

The methods of examination for dysosmia were reported. One thousand four hundred cases of dysosmia were treated in the last 5 years by the following examinations: 1) Olfactory acuity (detected and recognized) threshold test using T & T olfactometer. 2) Blast injection method using an applicator permeated with Alinamin solution. 3) Intravenous olfaction test using Alinamin solution. 4) Endoscopic observation of olfactory mucosa using an endoscope. The parts of lesions were diagnosed and various treatments were selected based on the results of these examinations. Nose drops of adrenocortical hormone were mainly used, while medications of peripheral vasodilator or medications for reducing of the mucosa were used in some cases. Many cases were effectively improved by the nose drops of adrenocortical hormone in Japanese patients. More than 70% of total cases were improved by these treatments.     

Familial nesidioblastosis: severe neonatal hypoglycemia in two families.

Severe neonatal hypoglycemia with pathologic findings of diffuse nesidoblastosis of the pancreas is described in five children of both sexes from two families with unaffected parents. This appears to represent an autosomal recessive disorder of pancreatic development. Despite extensive testing, the diagnosis of hyperinsulinism was difficult in the index case of each family and delayed definitive treatment. Medical therapy with steroids and diazoxide was unsuccessful; pancreatectomy was required to treat persistent hypoglycemia. An abnormality of circulating glucagon found in one child with this disorder suggested that hyperinsulinism may not be the sole hormonal imbalance present, but rather that this disease is one of generalized disturbance of islet cell function. The history of severe, persistent neonatal hypoglycemia in an older sibling should lead the physician to investigate subsequent children for the presence of asymptomatic hypoglycemia.     

Glutathione S-transferase Omega class (GSTO) polymorphisms in a sample from Rome (Central Italy)

Abstract

Glutathione S-transferases are a superfamily of enzymes that are involved in biotransformation of drugs, xenobiotics and play a fundamental role in the protection of cells from oxidative stress. In humans, the recently described GST Omega class contains two expressed genes GSTO1 and GSTO2, located on chromosome 10 (10q24.3). Four polymorphisms in GSTO genes have been identified in ethnic groups: GSTO1*A140D (rs4925), GSTO1*E155del (rs56204475), GSTO1*E208K (rs11509438) and GSTO2*N142D (rs156697). This study provides the allele frequencies of GSTO polymorphism in a sample consisting of 116 apparently healthy individuals of both sexes from Rome (Central Italy). Detection of GSTO1*A140D and GSTO2*N142D alleles was performed by PCR-RFLP analysis, while GSTO1*E155del and GSTO1*E208K alleles were detected using the Confronting Two-Pair Primers analysis (PCR-CTPP) and allele specific PCR, respectively. The GSTO allele frequencies found in the Italian sample were included in the variability range observed in European populations. Comparison between the data presented in this study and data in previous studies showed different patterns among European, Asian and African populations.     

心钠素对犬冠状动脉循环的调节作用

目的 :探讨生理及病理状态下心钠素调节冠状动脉循环的意义。方法 :应用测量冠状动脉直径的超声波晶体及测冠脉血流的多普勒探针 ,制成动物模型。将犬分别进行如下处理 :单独应用心钠素 ( 2 5、5 0和 1 0 0 ng· kg-1 · min-1 )持续静脉注射 ;HS- 1 4 2 - 1 ( HS)预处理后应用心钠素 ;单独应用 HS,测量冠状动脉直径、冠状动脉血流、平均动脉压、心率、血浆心钠素和环磷酸鸟苷浓度。结果 :单独应用心钠素可扩张冠状动脉直径 ,降低平均动脉压 ,对冠状动脉血流及心率无影响 ,增加血浆心钠素和环磷酸鸟苷浓度 ;HS处理后用心钠素 ,仅增加血浆心钠素浓度 ,其他指标均受抑制 ,单独用 HS对指标无影响。结论 :在以心功能不全为代表的病态下心钠素浓度升高 ,可通过直接扩张血管对维持冠脉循环起重要作用