Association of microRNAs genes polymorphisms with rheumatoid arthritis in Egyptian female patients

ObjectiveTo investigate whether miRNA-499 (rs3746444) and miRNA-146a (rs2910164) genes polymorphisms are independent factors for rheumatoid arthritis (RA) in Egyptians, and whether they influence disease severity and activity.MethodsTwo hundred and seventeen RA patients and 245 healthy controls were enrolled in this study. Polymorphisms of miRNA-146a and miRNA-499 genes were detected using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP).ResultsThe miRNA-499 CT genotype was an independent factor of RA. The miRNA-499 CT, CC genotypes and C allele frequencies were significantly increased in erosive RA group. Moreover, the heterozygote CT had more severe and more active form of the disease compared with homozygote CC or TT. However, we did not find any significant association of miRNA-146a polymorphism with RA risk, severity, and activity.ConclusionThe miRNA-499 polymorphism is an independent factor of RA, and influences disease severity and activity.     

BONE-INDUCTIVE EFFICACY OF RECOMBINANT HUMAN BONE MORPHOGENETIC PROTEIN-2 EXPRESSED IN ESCHERICHIA COLI: AN EXPERIMENTAL STUDY IN RAT MANDIBULAR DEFECTS

Because to our knowledge the efficacy of prokaryotically expressed recombinant human bone morphogenetic proteins (rhBMP) to promote orthotopic osteogenesis has not previously been investigated, our aim was to test the efficacy of rhBMP-2 produced in Escherichia coli to promote bone healing in a standardised experimental bone healing model in rat mandibles. Different doses of rhBMP-2 were delivered in an absorbable collagen sponge carrier, and microporous barrier membranes were placed over half the number of defects in each treatment group, thereby making intraosseous cells the only recruitment source for new osteogenic cells. Results were evaluated by computerised image analysis after 12 and 24 days. The relative efficacy of rhBMP-2 preparations of different purity was also compared. E coli-produced rhBMP-2 stimulated bone healing, but its efficacy was estimated to be about one order of magnitude less than that of rhBMP-2 expressed in eukaryotic cells. We conclude that bacterially expressed rhBMP-2 is osteogenic in vivo, although higher doses will be required than of rhBMP-2 expressed in mammalian cell lines.     

人CD147胞内结构域蛋白的表达与纯化

目的 构建人CD147胞内结构域（CD147 intracellular domain,CD147ICD）融合蛋白质粒,表达、提取、纯化CD147ICD蛋白并进行蛋白质互作分析。方法 将CD147ICD的cDNA片段插入pET-32a(+)质粒载体后,转化大肠杆菌Origami B(DE3)感受态细胞,大量培养后用异丙基硫代-β-d-半乳糖苷诱导蛋白表达,超声破碎法裂解细胞,采用Ni-Sepharose FF预装柱纯化TrxA-His6-CD147ICD融合蛋白,经凝血酶酶切、超滤后获得CD147ICD蛋白。结果 CD147ICD融合蛋白质粒经测序比对后表明构建成功;采用Ni-Sepharose FF预装柱纯化的TrxA-His6-CD147ICD融合蛋白纯度和浓度均较高,经凝血酶酶切、超滤后获得的CD147ICD蛋白纯度在95%以上,蛋白浓度为0.74 mg/mL。结论 纯化出了高纯度的、具有生物学活性的CD147ICD蛋白,为CD147ICD互作分子的鉴定及结构解析奠定了基础。     

Novel approaches to develop community-built biological network models for potential drug discovery

Introduction: Hundreds of thousands of data points are now routinely generated in clinical trials by molecular profiling and NGS technologies. A true translation of this data into knowledge is not possible without analysis and interpretation in a well-defined biology context. Currently, there are many public and commercial pathway tools and network models that can facilitate such analysis. At the same time, insights and knowledge that can be gained is highly dependent on the underlying biological content of these resources. Crowdsourcing can be employed to guarantee the accuracy and transparency of the biological content underlining the tools used to interpret rich molecular data.

Areas covered: In this review, the authors describe crowdsourcing in drug discovery. The focal point is the efforts that have successfully used the crowdsourcing approach to verify and augment pathway tools and biological network models. Technologies that enable the building of biological networks with the community are also described.

Expert opinion: A crowd of experts can be leveraged for the entire development process of biological network models, from ontologies to the evaluation of their mechanistic completeness. The ultimate goal is to facilitate biomarker discovery and personalized medicine by mechanistically explaining patients’ differences with respect to disease prevention, diagnosis, and therapy outcome.     

MicroRNA‐21 (miR‐21) expression in hypothermic machine perfusate may be predictive of early outcomes in kidney transplantation

Hypothermic machine perfusion is effective in improving outcome following kidney transplantation. Molecular analyses of hypothermic machine perfusate (HMP) have the potential to identify biomarkers of organ viability prior to transplantation, offering significant advantages to the transplant surgeon, and leading to a potential increase in the organ donor pool. MicroRNAs are emerging as important biomarkers in the context of kidney injury and transplantation. Recent data demonstrate increased microRNA‐21 (miR‐21) expression in the kidney following acute kidney injury. This study investigated the potential of miR‐21 detected in HMP to act as a sentinel for early kidney transplant outcomes. MiR‐21 was found to be readily detectable in HMP by RT‐qPCR. Eleven ECD kidneys were maintained on a hypothermic machine perfusion system for a median 627 (range 117–1027) minutes, and evaluation of flow and resistance characteristics suggested stability on the machine from 60 min post‐perfusion. MiR‐21 quantification at 60 min post‐perfusion correlated with eGFR at 6 and 12 months post‐transplantation. These data suggest that miR‐21 expression in HMP may be predictive of early outcomes following kidney transplantation. In the era of ECD kidneys, a reliable measure of organ quality is urgently needed, and this study suggests miR‐21 may be such a marker.     

MicroRNA在胃癌中的调控机制及预后价值

作为非编码RNA (ncRNAs)的一种,微小RNA(microRNA,miRNA)在基因表达调控方面受到广泛关注.MiRNA参与细胞周期、细胞凋亡、细胞迁移等进程,在肿瘤发生发展过程中也发挥重要作用.中国胃癌发病率较高,但对胃癌的治疗未能取得突破性进展,进展期胃癌患者往往预后不佳.目前尚缺乏可对胃癌患者预后进行判断的有效生物学标志,胃癌耐药也是临床的棘手问题.如何根据有效的生物学标志来判断预后,并针对这些靶点展开治疗,如何改善胃癌化疗耐药问题.miRNA在基因调控方面的作用给攻破这些难题带来了可能.该文是近年来关于miRNA与胃癌患者预后关系,改善患者预后,化疗耐药调控等方面的最新研究的综合论述.     

2型糖尿病肾病患者血清生长分化因子-15的表达及其临床意义

目的：探讨2型糖尿病肾病患者血清生长分化因子-15(GDF-15)的表达情况及临床意义。方法选择2011年12月至2015年2月在我院内分泌科进行治疗的2型糖尿病患者520例,根据尿白蛋白的含量水平分为正常白蛋白尿组(n=192)和微量白蛋白尿组(n=185)以及大量白蛋白尿组(n=143),应用酶联免疫吸附法(ELISA)检测并比较每组患者的血清GDF-15水平。结果正常白蛋白尿组患者血清GDF-15为(704.5±82.5) pg/ml,微量白蛋白尿组患者为(864.0±85.4) pg/ml,大量白蛋白尿组患者为(1773.9±113.5) pg/ml。正常白蛋白尿组和微量白蛋白尿组患者的血清GDF-15水平明显低于大量白蛋白尿组,差异均具有统计学意义(P<0.05);微量白蛋白尿组的血清GDF-15水平明显高于正常白蛋白尿组,差异具有统计学意义(P<0.05)。多元线形回归分析显示,微量白蛋白(mAlb)与白蛋白(Alb)呈负相关,与GDF-15呈正相关（P<0.05）。结论2型糖尿病肾病患者在临床的不同时刻血清GDF-15也会发生变化,患者血清GDF-15的高表达对病情的诊断和预后评价具有重要价值。     

放射性125I粒子对肺癌A549细胞裸鼠移植瘤VEGF表达的影响

目的 探讨放射性125I粒子对肺癌A549细胞裸鼠移植瘤血管内皮细胞生长因子(VEGF)表达的影响.方法 建立肺癌A549细胞裸鼠移植瘤模型,将24只小鼠随机分为对照组和观察组,每组12只.观察组小鼠植入125I粒子,对照组小鼠植入空白粒子.检测裸鼠移植瘤体积、瘤重,计算抑瘤率.采用免疫组织化学法检测移植瘤组织内VEGF蛋白表达.采用PCR法检测移植瘤组织内VEGF mRNA表达.结果 观察组小鼠移植瘤体积及瘤重[(0.658±0.213) g]均明显小于对照组[(1.807±0.625) g],组间比较差异均具有统计学意义(P<0.05);观察组小鼠平均抑瘤率为(68.2±5.3)%;观察组移植瘤中VEGF蛋白表达及VEGF mRNA表达分别为(22.6±5.9)、(98.7±8.2),均明显低于对照组的(58.5±6.4)、(138.7±5.2),组间比较差异均具有统计学意义(P<0.05).结论 放射性125I粒子植入可显著抑制肺癌A549细胞裸鼠移植瘤生长,抑制VEGF表达可能是其主要作用机制.     

抗核抗体和抗核抗体谱联合检测诊断自身免疫性疾病的临床价值

目的：探讨自身免疫性疾病(AID)患者血清抗核抗体(ANA)和抗核抗体谱(ANAs)的表达及临床意义。方法选取2013年1月至2015年4月海南省第三人民医院收治的194例AID患者和103例非AID患者(对照组),根据美国风湿病学会的诊断标准将194例AID患者分为系统性红斑狼疮(SLE)组、混合性结缔组织病(MCTD)组、干燥综合征(SS)组、硬皮病(PSS)组、类风湿性关节炎(RA)组和过敏性紫癜(AP)组。采用间接免疫荧光法(IIF)和免疫印迹法(IBT)检测所有患者的ANA和ANAs,并进行统计分析。结果女性患者检出ANA、Ul-SnRNP、抗dsDNA、抗SSA、抗SSB、抗核小体和抗组蛋白的阳性率均明显高于男性,差异均有统计学意义(P<0.05);在不同AID患者中ANA检测均呈阳性,其中SLE组阳性率最高,达93.5%,其次分别为MCTD组(85.0%)、SS组(79.2%)、PSS组(69.2%)、RA组(35.8%)、AP组(20.0%),均明显高于对照组(4.9%),差异均有统计学意义(P<0.05)。RA组主要表达抗组蛋白(47.7%),明显高于对照组(1.0%)。SLE组主要表达抗SmDl (37.9%)、抗dsDNA (66.1%)、抗SSA (41.9%）、抗核小体(77.4%)和抗组蛋白(37.1%),均明显高于对照组,差异均有统计学意义(P<0.05)。SS组主要表达抗SSA (58.3%)和抗SSB (50.0%),均明显高于对照组。MCTD组主要表达Ul-SnRNP (35.0%)、抗SSA (60.0%)和抗SSB (30.0%),均明显高于对照组。PSS组和AP组主要表达抗Scl-70(46.1%和10.0%),均明显高于对照组,差异均有统计学意义(P<0.05);130例ANA阳性患者检出97例ANAs阳性,符合率74.6%;64例ANA阴性患者检出58例ANAs阴性,符合率为90.6%;AID患者ANA核型分布以核颗粒型和核均质型为主,其他核型较少见;核颗粒型多为抗SSA (48.5%)、抗SSB (23.7%)和抗组蛋白(26.8%),核均质型多为抗dsDNA (28.9%)、抗核小体(29.9%)和抗组蛋白(25.7%)。结论 ANA可辅助诊断自身免疫性疾病,但缺乏特异性,联合抗核抗体谱检测对AID的诊断有重要意义。     

慢性心力衰竭患者血浆miRNA表达谱分析

目的：对慢性心力衰竭(CHF)患者的血浆miRNA进行二代测序,探讨CHF患者血浆中miRNA的表达差异。方法收集2013年7月至2015年6月间在北京大学深圳医院治疗的40例CHF患者和10例健康对照个体,应用第二代高通量测序技术对其血浆miRNA进行测序,寻找差异表达的miRNA。另收集同期CHF患者和健康对照个体各96例,用逆转录-实时荧光定量聚合酶链反应对差异表达的miRNA进行验证。结果与健康对照者相比,高通量测序显示在CHF患者的血浆中miR-184、miR-651-5p、miR-130b-5p、miR-203a-3p、miR-548e-3p和miR-106a-5p的表达水平存在显著上调,miR-31-5p和miR-5091的表达水平存在下调,逆转录-实时荧光定量聚合酶链反应检测血浆miRNA的表达差异与高通量测序的结果相符。结论 CHF患者的血浆miRNA与健康个体的表达谱存在差异,差异表达的miR-184、miR-651-5p、miR-130b-5p、miR-203a-3p、miR-548e-3p、miR-106a-5p、miR-31-5p和miR-5091或许与CHF的发生有关。