Pou4f2‐GFP knock‐in mouse line: A model for studying retinal ganglion cell development

Pou4f2 acts as a key node in the comprehensive and step‐wise gene regulatory network (GRN) and regulates the development of retinal ganglion cells (RGCs). Accordingly, deletion of Pou4f2 results in RGC axon defects and apoptosis. To investigate the GRN involved in RGC regeneration, we generated a mouse line with a POU4F2‐green fluorescent protein (GFP) fusion protein expressed in RGCs. Co‐localization of POU4F2 and GFP in the retina and brain of Pou4f2‐GFP/+ heterozygote mice was confirmed using immunofluorescence analysis. Compared with those in wild‐type mice, the expression patterns of POU4F2 and POU4F1 and the co‐expression patterns of ISL1 and POU4F2 were unaffected in Pou4f2‐GFP/GFP homozygote mice. Moreover, the quantification of RGCs showed no significant difference between Pou4f2‐GFP/GFP homozygote and wild‐type mice. These results demonstrated that the development of RGCs in Pou4f2‐GFP/GFP homozygote mice was the same as in wild‐type mice. Thus, the present Pou4f2‐GFP knock‐in mouse line is a useful tool for further studies on the differentiation and regeneration of RGCs.     

参麦注射液与布地奈德联合治疗急性酒精中毒伴吸入性肺炎的临床疗效

目的:探讨参麦注射液与布地奈德联合治疗急性酒精中毒伴吸入性肺炎的临床疗效及对肝功能和炎症指标的影响。方法:选择2014年3月至2015年8月在我院接受治疗的120例急性酒精中毒伴吸入性肺炎患者为研究对象,按照随机数字表法分为治疗组和对照组,每组60例患者,对照组给予布地奈德治疗,治疗组在对照组的基础上给予参麦注射液治疗,观察两组患者的临床疗效、症状消失时间、肝功能及血清肿瘤坏死因子-α、IL-6和IL-8炎性细胞因子的变化。结果:治疗组的总有效率为95.00%显著高于对照组的83.00%,差异有统计学意义(X~2=-4.227,P=0.039)。治疗组的哮鸣音、湿罗音、咳嗽和气喘消失时间明显短于对照组,差异有统计学意义(P0.005)。治疗组的肝功能ALT、AsT和GGT明显改善,炎性因子TNF-α、IL-6和IL-8水平下降明显,差异均有统计学意义(P0.005)。结论:参麦注射液与布地奈德联合治疗急性酒精中毒伴吸入性肺炎疗效确切,可以有效缩短患者的症状消失时间,改善肝功能,减轻炎症反应,值得临床推广应用。     

不同时间高压氧治疗对颅脑损伤患者认知功能障碍的影响

目的:探讨不同时间高压氧治疗对颅脑损伤患者认知功能障碍的影响。方法:选择了2006年5月-2015年5月在我院接受高压氧治疗的500例颅脑损伤手术或非手术患者,根据患者开始采取高压氧治疗时间的不同分为A(入院后7d内)、B(7-30 d)、C(30 d后)三组,通过相应的治疗比较三组患者认知功能评分、FIM评分及治疗效果。结果:高压氧治疗后,三组患者较治疗前都有了不同程度上的改善,但A组在定向能力、专注能力、理解能力、复述能力、结构组织能力、记忆能力、计算能力以及自我照顾、转移、行走、交流、社会认知较B组、C组改善显著(P0.05),而B组与C组间认知功能各项评分、FIM评分比较差异无统计学意义(P0.05);A组患者治疗后总有效率为90.63%,显著优于B组73.89%的总有效率及C组70.00%的总有效率(P0.05),而B组与C组比较差异无统计学意义(P0.05)。结论:颅脑伤患者尽早的接受高压氧治疗其认知功能改善越明显,治疗效果越好。     

丹参酮ⅡA 磺酸钠注射液对原发性高血压患者血脂水平及内皮功能的影响

目的:研究丹参酮ⅡA磺酸钠注射液对原发性高血压患者血脂水平及内皮功能的影响。方法:选择2013年5月-2015年10月在我院首次诊断为原发性高血压的80例患者作为研究对象,根据治疗方法不同将入组患者随机分为实验组和对照组。实验组患者接受丹参酮ⅡA磺酸钠注射液联合口服降压药物治疗,对照组患者仅接受口服降压药物治疗,比较两者患者治疗前后的血压、肝肾功能、血脂水平以及内皮功能指标的变化。结果:治疗后,两组患者的收缩压、舒张压水平均低于治疗前,且实验组患者的收缩压、舒张压水平与对照组比较均无统计学差异。治疗后,两组患者的ALT、AST、Scr水平均与治疗前比较均无显著差异,且实验组患者的ALT、AST、Scr水平与对照组比较无统计学差异。治疗后,实验组患者的TC、TG、LDL水平明显低于治疗前(P0.05),对照组患者的TC、TG、LDL与治疗前比较无统计学差异(P0.05),实验组患者的TC、TG、LDL水平显著低于对照组(P0.05)。结论:丹参酮ⅡA磺酸钠注射液有助于改善原发性高血压患者的血脂代谢以及内皮功能,且对患者的肝肾功能无明显影响。     

纤维支气管镜对急诊重症肺炎合并呼吸衰竭患者呼吸功能及血清炎症因子水平的影响

目的:探讨纤维支气管镜对急诊重症肺炎合并呼吸衰竭患者呼吸功能及炎症反应的影响。方法:选择2013年9月-2015年9月在我院接受治疗的重症肺炎合并呼吸衰竭患者89例作为研究对象,根据治疗方法不同,将患者分为研究组(47例)与对照组(42例)。对照组患者采用常规抗感染治疗,研究组患者在对照组基础上采用纤维支气管镜肺泡灌洗治疗。观察并比较两组患者治疗前后的动态顺应性(Cdyn)、氧合指数(Pa O2/Fi O2)、呼吸做功(WOB)、血清炎症因子水平的变化情况。结果:治疗前,两组患者Cdyn,WOB,Pa O2/Fi O2,CD11b+中性粒细胞、STREM-1及HMGB-1水平比较,差异均无统计学意义(P0.05);治疗后,对照组患者Cdyn、Pa O2/Fi O2及WOB均较治疗前显著升高,差异具有统计学意义(P0.05);研究组患者Cdyn及Pa O2/Fi O2较治疗前显著升高,而WOB较治疗前显著降低,差异具有统计学意义(P0.05);研究组患者Cdyn及Pa O2/Fi O2高于对照组,而WOB低于对照组,差异具有统计学意义(P0.05);两组患者CD11b+中性粒细胞、STREM-1及HMGB-1水平均较治疗前显著降低,且研究组显著低于对照组,差异均具有统计学意义(P0.05)。结论:纤维支气管镜肺泡灌洗能够改善急诊重症肺炎合并呼吸衰竭患者的呼吸功能,缓解机体炎症反应,具有重要的临床意义。     

网络云交互技术辅助外科实践教学的创新应用探索

目的:探索传统教学模式结合网络云交互技术在外科实践教学中的应用效果。方法:于我校2011级五年制临床医学专业本科生中选取76人,随机平均分为两组,分别采取单纯传统大课教学模式、传统教学结合网络云交互技术教学模式;课程结束后进行闭卷理论考试和实践操作考试,综合两项成绩进行评定。同时向两组学员发放调查问卷,以此评价使用网络云交互技术辅助教学的效果。结果:采取传统教学结合网络云交互技术教学模式组的成绩优于采取单纯传统教学模式组,两组成绩差异具有统计学意义(P0.05)。结论:在传统教学模式的基础上添加网络云交互技术更能激发学生的学习兴趣,大大提高了学习效率,更有利于本科生外科实践的教学以及教学资源的整合及教学档案的建立。     

血管内皮功能与急性脑梗死患者梗死类型及颈动脉斑块性质的相关性

目的:研究急性脑梗死(ACI)患者血管内皮功能的变化情况,并分析其与脑梗死类型及颈动脉斑块性质的相关性。方法:选择2014年6月到2015年6月在我院收治的脑梗死患者100例作为研究对象。根据病史及磁共振成像(MRI)诊断结果分为ACI组(n=70)和非急性脑梗死(NACI)组(n=30),对两组患者间的血管内皮细胞功能的相关指标进行比较,同时按照梗死类型以及颈动脉斑块性质不同对ACI组进行分组,探究梗死类型与斑块性质与反应性充血指数(RHI)的相关性。结果:ACI组纤维蛋白原(Fbg)、低密度脂蛋白(LDL)、大内皮素-1(big ET-1)、超敏C反应蛋白(hs-CRP)和内皮细胞生长因子(VEGF)水平均高于NACI组,RHI水平低于NACI组,差异有统计学意义(P0.05)。不同急性脑梗死TOAST分型中心源性栓塞型(CE)、小血管闭塞或腔隙性梗死型(SAA)、不明原因型(SUE)和大动脉粥样硬化型(LAA)测得RHI水平逐渐上升,hs-CRP水平和big ET-1水平逐渐下降,差异具有统计学意义(P0.05);比较不同斑块性质发现无斑块组、硬斑块组、软斑块组和混合斑块组的VEGF水平逐渐升高,RHI水平逐渐降低,差异均有统计学意义(P0.05)。相关性分析显示,RHI与斑块性质、hs-CRP以及big ET-1之间呈现负相关(r=-0.672,-0.402,-0.512,P0.05)。结论:血管内皮功能与急性脑梗死类型和颈动脉斑块性质有相关性,可作为评估梗死类型和斑块性质的预测指标。     

Investigating core genetic-and-epigenetic cell cycle networks for stemness and carcinogenic mechanisms,and cancer drug design using big database mining and genome-wide next-generation sequencing data

Recent studies have demonstrated that cell cycle plays a central role in development and carcinogenesis. Thus, the use of big databases and genome-wide high-throughput data to unravel the genetic and epigenetic mechanisms underlying cell cycle progression in stem cells and cancer cells is a matter of considerable interest.

Real genetic-and-epigenetic cell cycle networks (GECNs) of embryonic stem cells (ESCs) and HeLa cancer cells were constructed by applying system modeling, system identification, and big database mining to genome-wide next-generation sequencing data. Real GECNs were then reduced to core GECNs of HeLa cells and ESCs by applying principal genome-wide network projection. In this study, we investigated potential carcinogenic and stemness mechanisms for systems cancer drug design by identifying common core and specific GECNs between HeLa cells and ESCs. Integrating drug database information with the specific GECNs of HeLa cells could lead to identification of multiple drugs for cervical cancer treatment with minimal side-effects on the genes in the common core. We found that dysregulation of miR-29C, miR-34A, miR-98, and miR-215; and methylation of ANKRD1, ARID5B, CDCA2, PIF1, STAMBPL1, TROAP, ZNF165, and HIST1H2AJ in HeLa cells could result in cell proliferation and anti-apoptosis through NFκB, TGF-β, and PI3K pathways. We also identified 3 drugs, methotrexate, quercetin, and mimosine, which repressed the activated cell cycle genes, ARID5B, STK17B, and CCL2, in HeLa cells with minimal side-effects.     

Diverse mechanisms of metaeffector activity in an intracellular bacterial pathogen,Legionella pneumophila

Pathogens deliver complex arsenals of translocated effector proteins to host cells during infection, but the extent to which these proteins are regulated once inside the eukaryotic cell remains poorly defined. Among all bacterial pathogens, Legionella pneumophila maintains the largest known set of translocated substrates, delivering over 300 proteins to the host cell via its Type IVB, Icm/Dot translocation system. Backed by a few notable examples of effector–effector regulation in L. pneumophila, we sought to define the extent of this phenomenon through a systematic analysis of effector–effector functional interaction. We used Saccharomyces cerevisiae, an established proxy for the eukaryotic host, to query > 108,000 pairwise genetic interactions between two compatible expression libraries of ~330 L. pneumophila‐translocated substrates. While capturing all known examples of effector–effector suppression, we identify fourteen novel translocated substrates that suppress the activity of other bacterial effectors and one pair with synergistic activities. In at least nine instances, this regulation is direct—a hallmark of an emerging class of proteins called metaeffectors, or “effectors of effectors”. Through detailed structural and functional analysis, we show that metaeffector activity derives from a diverse range of mechanisms, shapes evolution, and can be used to reveal important aspects of each cognate effector's function. Metaeffectors, along with other, indirect, forms of effector–effector modulation, may be a common feature of many intracellular pathogens—with unrealized potential to inform our understanding of how pathogens regulate their interactions with the host cell.     

Structural mutation analysis of PTEN and its genotype‐phenotype correlations in endometriosis and cancer

The phosphatase and tensin homolog deleted on chromosome ten (PTEN) gene encodes a tumor suppressor phosphatase that has recently been found to be frequently mutated in patients with endometriosis, endometrial cancer and ovarian cancer. Here, we present the first computational analysis of 13 somatic missense PTEN mutations associated with these phenotypes. We found that a majority of the mutations are associated in conserved positions within the active site and are clustered within the signature motif, which contain residues that play a crucial role in loop conformation and are essential for catalysis. In silico analyses were utilized to identify the putative effects of these mutations. In addition, coarse‐grained models of both wild‐type (WT) PTEN and mutants were constructed using elastic network models to explore the interplay of the structural and global dynamic effects that the mutations have on the relationship between genotype and phenotype. The effects of the mutations reveal that the local structure and interactions affect polarity, protein structure stability, electrostatic surface potential, and global dynamics of the protein. Our results offer new insight into the role in which PTEN missense mutations contribute to the molecular mechanism and genotypic‐phenotypic correlation of endometriosis, endometrial cancer, and ovarian cancer. Proteins 2016; 84:1625–1643. © 2016 Wiley Periodicals, Inc.