Regulator of G-protein signaling 2 inhibits acid-induced mucin5AC hypersecretion in human airway epithelial cells

Mucus hypersecretion is a common pathological feature of inflammatory airway diseases. Previous studies have shown that acidic microenvironment of inflamed airways may provoke the pathophysiology of inflammatory airway diseases. However, the acidic-sensing and negative regulatory mechanisms that mediate mucus hypersecretion in inflamed airway remain elusive. Thus, we sought to explore the role of ovarian cancer G-protein-coupled receptor 1 (OGR1) in acid-induced mucin5AC (MUC5AC) hypersecretion in human airway epithelium and the inhibitory effect of regulator of G-protein signaling 2 (RGS2) in this process. We found that airway acidification increased [Ca²⁺]_i, which was required for MUC5AC secretion. Knocking-down OGR1 and G_q with siRNAs and pretreating cells with phospholipase C inhibitor effectively attenuated acid-induced cellular responses. Moreover, the overexpression of wild-type RGS2 attenuated acid-induced cellular responses. In contrast, reducing RGS2 with siRNA enhanced the increases in acid-induced cellular responses. These data suggest that airway acidification can induce MUC5AC hypersecretion through an OGR1-mediated mechanism and RGS2 can inhibit acid-induced MUC5AC hypersecretion at OGR1 receptor level.     

PPARγ inhibits inflammation and RANKL expression in epoxy resin-based sealer-induced osteoblast precursor cells E1 cells

ObjectivesThe AH26 of epoxy resin-based sealer is used widely owing to its excellent physical characteristics but it induces oxidative stress and cytotoxicity at the periapical tissues. AH26 exhibited cytotoxicity towards MC-3T3-E1 cells, which resulted in mitochondria-mediated apoptosis. Peroxisome proliferator-activated receptor (PPARγ) has an anti-inflammatory effect in several tissue and cells, but its action of AH26-related inflammation is not completely understood. The aim of this study is to investigate the anti-inflammatory and anti-osteoclastic mechanisms of PPARγ in AH26-induced MC-3T3 E1 cells.MethodsAH26 was prepared according to the manufacturer's instructions. The 1-day extraction sample, which was diluted by 30%, was tested in this experiment. Recombinant deficiency adenoviral PPARγ (Ad/PPARγ) was used to examine PPARγ over-expression in MC-3T3 E1 cells. AH26-induced reactive oxygen species (ROS) formation was analysed using 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) with fluorescence-activated cell sorting (FACS), and the expression of receptor activator of nuclear factor-κB ligand (RANKL) and inflammatory molecules was determined by immunoblotting. The anti-inflammatory and anti-osteoclastic mechanisms of the PPARγ-involved signal pathway was examined by immunoblotting.ResultsThe AH26 elutes induced inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), RANKL expression and ROS formation. In addition, the AH26 elutes suppressed the expression of PPARγ. However, the recovery of PPARγ expression with Ad/PPARγ resulted in the inhibition of iNOS, COX-2, RANKL and ROS formation despite the AH26 treatment in MC-3T3 E1 cells. The mechanism of PPARγ was confirmed by the blocking of nuclear factor kappa B (NF-κB) translocation to the nucleus after the suppression of ERK1/2, SAPK/JNK and AP-1 in AH26-induced MC-3T3 E1 cells.ConclusionFrom this result, PPARγ acts to inhibit bone destruction in AH26-induced bone cells. Therefore, the anti-inflammatory and anti-osteoclastic character of PPARγ might be applicable for healing periapical lesions more rapidly or reducing the induction of cellular inflammation caused by some endodontic sealers.     

A novel method of augmenting gene expression and angiogenesis in the normal and ischemic canine myocardium

This study presents a novel method that direct intramyocardial injection of low-dose plasmid DNA and microbubbles combined with insonation could further augment gene expression in normal and ischemic canine myocardium. Plasmids encoding enhanced green fluorescent protein (pEGFP) and hepatocyte growth factor (pHGF) (500???g) were individually mixed with 0.5?ml of microbubble solution (MB) and injected into the normal or acute ischemic canine myocardium. The dogs in the plasmid?+?MB/US group underwent insonation (US). Other dogs were randomly divided into three treatment groups: plasmid and insonation, plasmid and MB injection, and plasmid injection only. The EGFP and HGF mRNA expressions were assessed in the myocardium at the injection site and at sites 0.5 and 1?cm remote from the injection site. Compared to plasmid transfer alone, a mean 13.4-fold enhancement of gene expression was achieved in the EGFP?+?MB/US group at 48?h (p?<?0.01). HGF mRNA expression in ischemic zones was markedly elevated after 28?days, with a mean 9.0-fold enhancement in the HGF?+?MB/US group (p?<?0.01). EGFP protein expression was detected in the normal myocardium at 1?cm remote from the injection site in the EGFP?+?MB/US group. Similarly, HGF protein expression was detected in the ischemic myocardium at 0.5?cm remote from the injection site in the HGF?+?MB/US group. These findings indicate that the radius of gene expression was partly extended in the two plasmid?+?MB/US groups. The capillary density increased from 20.9?±?5.3/mm² in control myocardial infarction dogs without treatment to 126.7?±?38.2/mm² in the HGF?+?MB/US group (p?<?0.01). Taken together, the present data demonstrate that direct intramyocardial injection of an angiogenic gene and microbubbles combined with insonation can augment gene expression and angiogenesis. Consequently, this strategy may be a useful tool for gene therapy of ischemic heart disease.     

QRS波时限对起搏器依赖患者心脏功能的影响

目的:观察右心室起搏QRS波时限对起搏器依赖患者心脏功能的影响。方法:选取在我院诊断为Ⅲ度房室传导阻滞并行永久性右心室起搏的患者112例,以起搏QRS波时限将患者分为A组(起搏QRS波时限<190ms)和B组(起搏QRS波时限≥190ms),并对每例患者进行临床评估和心脏彩色多普勒超声检查,动态随访心脏功能,以随访期间出现明显心功能下降为终点,观察起搏QRS波时限与心脏功能的关系。同时根据随访期间是否出现心功能下降将患者分为心功能下降组(Y组)和心功能无下降组(N组),并进行各因素与心功能下降之间的单因素分析,取P<0.1的因素纳入Logistic回归模型,寻找影响起搏器依赖患者心功能的因素。结果:平均随访(45.46±23.00)个月,40例(28.57%)出现明显心功能下降,其中,A组24例(27.27%),B组16例(66.67%),2组间差异有统计学意义(P=0.0004),Y组的起搏QRS波时限较N组明显延长[(176.58±22.71)∶(159.74±20.23)ms,P<0.0001]。多元Logistic逐步回归分析结果显示,左心室舒张末期内径增大、射血分数下降、左束支传导阻滞、起搏QRS波时限≥190ms、年龄及起搏时间是起搏器依赖患者心功能下降的危险因素。结论:起搏QRS波时限延长是心脏功能下降和心力衰竭发生的危险因素,可以作为起搏器依赖患者起搏后心脏功能下降的预测指标。除此之外,左心室增大、左束支传导阻滞、射血分数降低、年龄及起搏时间也是起搏器依赖患者心脏功能下降的危险因素。     

活体肝移植后小肝综合征的防治

活体肝移植是治疗终末期肝病最有效的手段,但小肝综合征是成人活体肝移植术后一种发生率和病死率都较高的临床并发症,是制约成人间活体肝移植的主要原因之一.目前,如何有效防治小肝综合征已成为肝移植领域的研究热点.本篇就活体肝移植后小肝综合征的病理生理、危险因素及防治策略的最新进展作一简要综述.     

吻合口溃疡穿透致吻合口横结肠瘘1例

吻合口溃疡穿透导致吻合口横结肠瘘临床少见,表现无特异性,主要为腹痛、腹泻未消化物、呕吐粪样物、营养不良,胃肠镜检查及消化道造影有助于诊断。外科手术是唯一有效的治疗方式。     

右房峡部的影像解剖特征研究进展

右房峡部已被证实为典型心房扑动的关键传导部位,全程线性透壁消融右房峡部是临床介入治疗典型房扑的首选策略;但右房峡部内成分构成复杂,其解剖特点是影响消融结果与预后的重要因素。近年来,随着多维超声、高分辨率计算机体层成像和磁共振在心脏成像的广泛应用和大量相关临床研究的开展,右房峡部长度增加、存在欧氏嵴以及过深的隐窝和囊袋、与下腔静脉成角增加等解剖变异均可能增加导管消融的难度,对个体而言,舒张状态下的右房峡部长度是影响消融成功率的重要因素。现就此研究进展作出综述。     

蛋白酶抑制剂联合聚乙二醇干扰素及利巴韦林治疗慢性丙型肝炎初治患者疗效及安全性的荟萃分析

目的 比较蛋白酶抑制剂(特拉普韦或波塞普韦)联合聚乙二醇干扰素(Peg-IFN)及利巴韦林(RBV)的三联治疗方案与Peg-IFN联合RBV的标准二联方案治疗基因1型初治慢性丙型肝炎的疗效及安全性.方法 检索PubMed、EMBASE、OVID和Cochrane图书馆等数据库,纳入比较蛋白酶抑制剂联合Peg-IFN及RBV的三联治疗方案与Peg-IFN联合RBV的标准二联方案治疗基因1型初治慢性丙型肝炎疗效与安全性的随机对照试验.检索词为protease inhibitor、hepatitis C和genotype 1.主要的研究结果包括:持续病毒学应答率(SVR)、复发率、贫血发生率、因严重不良反应而致的停药率.采用比值比(OR)及其95％可信区间(CI)作为研究结果的评价指标,异质性通过x2检验及I2检验评价,异质性明显时采用随机效应模式,否则采用固定效应模式.结果 共纳入5篇文献,包括3200例患者.三联治疗组的SVR率高于标准二联组(65.4％比40.9％,OR=2.92,95％口CI为2.5～3.42,P＜0.01),复发率低于二联治疗组(11.3％比24.8％,OR=0.42,95％CI为0.26～0.68,P＜0.01),贫血发生率高于二联治疗组(44.1％比26.2％,OR=2.25,95％ CI为1.9～2.65,P＜0.01),因严重不良反应而致的停药率也高于二联治疗组(12.4％比7.7％,OR=1.66,95％ CI为1.19～2.32,P＜0.01).亚组分析结果显示,三联治疗组(包括24或28周组、48周组、应答指导疗程组)均有较高的SVR率及较低的复发率.结论 特拉普韦或波塞普韦联合Peg-IFN及RBV的三联治疗方案对于基因1型的初治慢性丙型肝炎的疗效优于Peg-IFN联合RBV的二联治疗方案,但增加了贫血发生率及因不良反应而导致的停药率.     

Characterization of gastric cancer models from different cell lines orthotopically constructed using improved implantation techniques

AIM: To develop orthotopic gastric cancer mouse models from different cell lines and characterize the tumor features to assist further in preclinical trials and clinical treatment strategies.METHODS: Human gastric cancer SGC-7901 and BGC-823 cell suspensions were injected subcutaneously into nude mice to develop solid tumors, and tumor tissue pieces were then implanted under the serous coat of the stomach. An autopsy was performed on all animals of the SGC-7901 and BGC-823 models to observe the primary tumor growth and metastases using pathological and immunohistochemical methods.RESULTS: Both models showed large tumors in situ resulting in pressure and infiltration of the adjacent organs. The gastric cavity became smaller, along with stenosis of the cardia or pylorus. There were biological and statistical differences between the two models. The metastasis rate in involved organs (lymph nodes, kidney, spleen, testis) was significantly higher in the BGC-823 model compared to the SGC-7901 model (P < 0.05 or P < 0.01). The median survival of the BGC-823 model was shorter than that of SGC-7901 (23 d vs 84 d, P < 0.05). Histopathologically, the primary tumor and metastatic lesions of the two models showed obvious atypia and mucus in the cytoplasm. Compared with the SGC-7901 model, BGC-823 appeared more poorly differentiated (absence of adenoid structure), had a smaller volume, and richer capillary structure. Immunohistochemical staining revealed cytokeratin 20 and epithelial membrane antigen expression was positive in the SGC-7901 tumors, while negative in BGC-823 ones.CONCLUSION: Models using the SGC-7901 and BGC-823 cell lines were established which could function in gastric cancer research on carcinogenesis mechanism and drug discovery. The two models showed different tumor behavior and the latter was more malignant than the former.     

丹参、人参及蛤蚧组方对大鼠肺纤维化凋亡基因作用的研究

目的 研究丹参、人参及蛤蚧组方对博莱霉素致大鼠肺间质纤维化模型凋亡基因的表达,探讨该组方抗肺纤维化的作用机制.方法 SD雄性大鼠50只随机分为对照组、模型组、醋酸泼尼松组、治疗Ⅰ组(低剂量组:丹参83 mg/kg、人参50mg/kg及蛤蚧50mg/kg)及治疗Ⅱ组(高剂量组:丹参166mg/kg、人参50mg/kg及蛤蚧50 mg/kg);于第28天处死大鼠,RT-PCR 法检测大鼠肺组织bax及bcl-2基因mRNA表达水平,用单克隆抗体免疫组化法检测bax及bcl-2蛋白含量.结果 bax基因mRNA及蛋白在模型组、醋酸泼尼松组、治疗Ⅰ组及治疗Ⅱ组的表达明显升高,与对照组相比差异有统计学意义(P＜0.05);同时模型组升高幅度明显,治疗Ⅱ组升高幅度较低,两者相比差异有统计学意义(P ＜0.01);bcl-2基因mRNA及蛋白在醋酸泼尼松组、治疗Ⅰ组及治疗Ⅱ组的表达升高,并治疗Ⅱ组升高更明显,与模型组相比差异有统计学意义(P＜0.05).结论 丹参、人参及蛤蚧组方能通过调节细胞凋亡,下调bax活性,增强Bcl-2的活性,抑制细胞凋亡,减少纤维积聚和纤维化形成.