特异性抑制单核细胞中EMMPRIN基因表达的siRNA筛选和鉴定

目的 设计合成干扰细胞外基质金属蛋白酶诱导因子(EMMPRIN)基因表达的不同小型干扰RNA(siRNA),筛选出能高效抑制单核/巨噬细胞中EMMPRIN表达的siRNA.方法 根据人源EMMPRIN mRNA的序列,设计合成3对不同的且能干扰EMMPRIN基因表达的siRNA,将其转染THP-1单核细胞株,采用荧光定量PCR和蛋白印迹法评价基因表达情况.结果 设计合成的3对siRNA中的2对siRNA可以特异性抑制单核细胞中EMMPRIN mRNA和蛋白表达(分别减少70%和50%,P＜0.05).结论 成功设计合成了针对EMMPRIN基因的siRNA,并从中筛选出能特异且高效阻断EMMPRIN表达的siRNA,为进一步研究EMMPRIN基因对动脉粥样硬化形成的影响及其临床应用奠定了基础.     

肥大细胞类胰蛋白酶在其相关疾病中作用的研究

作为近年来备受关注的一种炎症介质，肥大细胞类胰蛋白酶有着多重生物活性效应，本文主要综述了肥大细胞类胰蛋白酶在其相关疾病发病机制中所起的重要作用；而对类胰蛋白酶的分泌和活性进行调控将有望成为此类相关疾病治疗的新途径。     

新型抗肿瘤药物双（β-二酮）钛（Ⅳ）配合物的合成

目的开发新型抗肿瘤药物。方法以苯甲酰丙酮、二硫化碳、1,2-二溴乙烷为起始原料,设计合成了两个新型含硫杂环双（β-二酮）配体（Ⅰa-Ⅰb）及其钛配合物（Ⅱa-Ⅱb）。结果合成得到的新型含硫杂环双（β-二酮）配体（Ⅰa-Ⅰb）及其钛配合物（Ⅱa-Ⅱb）通过元素分析、红外光谱1、HNMR、MS等手段对其进行了结构表征。结论此合成路线完全可行。     

同时稳定抑制X连锁凋亡抑制蛋白和生存素表达对胰腺癌细胞上皮-间质转化及侵袭性的影响

目的:探讨同时抑制X连锁凋亡抑制蛋白(XIAP)和生存素(survivin)后,对胰腺癌Panc-1细胞上皮-间质转化(EMT)及侵袭性的影响,并初步探讨其机制。方法:在前期实验构建的胰腺癌Panc-1-XS细胞(XIAP和survivin同时稳定抑制)中,运用Transwell小室实验及划痕实验分别检测细胞侵袭和迁移能力;半定量Western印迹分别检测钙黏蛋白-E(E-cadherin,上皮标志物)、锌指转录因子(Slug)蛋白(间质标志物)及第10号染色体同源丢失性磷酸酶——张力蛋白基因(PTEN)和磷酸化蛋白激酶B(P-Akt)蛋白的表达情况。结果:Panc-1-XS细胞侵袭和迁移能力显著下降,同时伴随E-cadherin蛋白表达显著上调及Slug蛋白显著下调,即出现间质-上皮转化(MET);PTEN蛋白表达上调、P-Akt蛋白表达下调。结论:同时抑制XIAP和survivin表达,能部分逆转胰腺癌Panc-1细胞EMT表型,显著减弱其侵袭和迁移能力;此调控过程可能通过PTEN/PI3K/Akt途径实现。     

内毒素脂多糖对胰岛细胞株INS-1细胞活力及胰岛素分泌的影响

目的 观察Toll样受体4(TLR4)在胰岛细胞株(INS-1)中的表达情况,探讨内毒素脂多糖(LPS)对INS-1细胞活力、胰岛素分泌功能以及TLR4表达的影响.方法 采用RT-PCR技术和Western blotting方法检测INS-1细胞和经1 mg/L LPS刺激的INS-1细胞中TLR4 mRNA和蛋白的表达变化;分别用不同质量浓度的LPS (0.01、0.1、1、5、10 mg/L)刺激INS-1细胞,CCK8法检测细胞活力,GSIS法测定胰岛素分泌功能.结果 RT-PCR和Western blotting检测结果显示:LPS刺激使INS-1细胞TLR4mRNA和蛋白的表达显著增加(P＜0.05).当LPS在0.1～ 10 mg/L时,INS-1细胞活力受到抑制,且呈剂量时间依赖性(P＜0.05);1 mg/L LPS刺激可使高糖环境下的INS-1细胞的胰岛素分泌量显著减少(P＜0.05).结论 一定浓度范围内的LPS刺激可抑制INS-1细胞活力,并减少高糖培养条件下细胞胰岛素的分泌量.LPS刺激能上调INS-1细胞TLR4 mRNA和蛋白的表达.内毒素可能通过直接作用损伤胰岛β细胞.     

红外/红光治疗仪缓解慢性软组织损伤症状的临床试验

[目的] 评价迈能MPET800红外/红光治疗仪,缓解慢性软组织损伤引起疼痛等症状的有效性及安全性.[方法] 将66名患者随机分为两组,对照组使用CQ-61型红外线治疗器,理疗14 d后评价临床疗效.[结果] 试验组综合疗效及局部疼痛疗效有效率优于对照组,压痛、局部肿胀及功能障碍疗效有效率与对照组无差异.[结论] 本仪器可缓解慢性软组织损伤引起的局部疼痛、压痛、功能障碍、局部肿胀症状.     

LINC00240 knockdown inhibits nasopharyngeal carcinoma progress by targeting miR‐26a‐5p

ObjectiveThis study intended to explore the regulatory functions of LINC00240 on nasopharyngeal carcinoma (NPC).MethodsMiR‐26a‐5p inhibitor, mimic, and siLINC00240 were transfected into NPC cells. QRT‐PCR was employed to assess miR‐26a‐5p and LINC00240 expressions. The targeting relationship of LINC00240 and miR‐26a‐5p was analyzed through dual luciferase reporter and RNA immunoprecipitation assay. Cell counting kit‐8 assay, colony formation assay, flow cytometry assay, wound healing assay, Transwell assay and in vitro angiogenesis assay were adopted for the evaluation of the effects of LINC00240 or miR‐26a‐5p and LINC00240 on NPC cells regarding cell proliferation, apoptosis and cycle, migration, invasion, and angiogenesis. EZH2, cell cycle, and epithelial‐mesenchymal transition (EMT)‐related protein expression was tested through Western blot.ResultsLINC00240 had a high expression in NPC tissues and cell lines. Silenced LINC00240 significantly suppressed the 5‐8F and HK1 cell proliferation, invasion, migration, and angiogenesis, but raised cell apoptosis, and cells were blocked in G0/G1 phase. MiR‐26a‐5p was a target of LINC00240. MiR‐26a‐5p upregulation suppressed the NPC cell proliferation, migration, invasion, angiogenesis, N‐cadherin and EZH2 expression, while it elevated apoptosis and p21, p27 and E‐cadherin expressions, whereas miR‐26a‐5p downregulation performed conversely. LINC00240 knockdown partially offset the effects of miR‐26a‐5p downregulation on cell proliferation, migration, invasion, angiogenesis, apoptosis, and EZH2.ConclusionLINC00240 knockdown restrained cell proliferation, invasion, migration, and angiogenesis, while it advanced apoptosis via miR‐26a‐5p in NPC by EZH2 inhibition.     

Effect of L-Glutamine on Chylomicron Formation and Fat-Induced Activation of Intestinal Mucosal Mast Cells in Sprague-Dawley Rats

Glutamine (Gln) is required for intestinal mucosal homeostasis, and it can promote triglyceride absorption. The intestinal mucosal mast cells (MMCs) are activated during fat absorption. This study investigated the potential role of Gln on fat absorption-induced activation of MMCs in rats. Lymph fistula rats (n = 24) were studied after an overnight recovery with the infusion of saline only, saline plus 85 mM L-glutamine (L-Gln) or 85 mM D-glutamine (D-Gln), respectively. On the test day, rats (n = 8/group) were given an intraduodenal bolus of 20% Intralipid contained either saline only (vehicle group), 85 mM L-Gln (L-Gln group), or 85 mM D-Gln (D-Gln group). Lymph was collected hourly for up to 6 h for analyses. The results showed that intestinal lymph from rats given L-Gln had increased levels of apolipoprotein B (ApoB) and A-I (ApoA-I), concomitant with an increased spectrum of smaller chylomicron particles. Unexpectedly, L-Gln also increased levels of rat mucosal mast cell protease II (RMCPII), as well as histamine and prostaglandin D₂ (PGD₂) in response to dietary lipid. However, these effects were not observed in rats treated with 85 mM of the stereoisomer D-Gln. Our results showed that L-glutamine could specifically activate MMCs to degranulate and release MMC mediators to the lymph during fat absorption. This observation is potentially important clinically since L-glutamine is often used to promote gut health and repair leaky gut.     

Microstructure Evolution and Deformation Mechanisms of As-Cast Antibacterial Ti6Al4V-5Cu Alloy for Isothermal Forging Process

The hot workability behavior of antibacterial Ti6Al4V-5Cu alloy was investigated using a hot compression experiment in the temperature range of 790–1040 °C and strain rate of 10⁻³–10 s⁻¹ with a strain of 0.4. The deformation behavior of the alloy was characterized by Gleeble 3800 compression experiment, and the relationship among deformed microstructures and deformation parameters was established. The deformations of Ti6Al4V-5Cu alloy were temperature and strain rate-dependent. Higher temperature and lower strain rate made power dissipation efficiency (η) increase and reach 89%. The activation energies (Q) in the dual-phase (α + β) and single β phase regions were calculated as 175.43 and 159.03 kJ mol⁻¹, respectively. In the dual (α + β) phase region, with an increase in strain rate, flow-softening behavior was dominated, however in the single β phase region such as processing at 940 °C. Flow stress increased slightly in which work-hardening behavior was dominated (especially between strain rates of 10⁻³–1 s⁻¹). The deformation at various conditions exhibited different stress-strain profiles, providing an insight that work hardening and flow softening coexisted in Ti6Al4V-5Cu alloy. The relative intensity of oscillatory change in flow stress profile decreased as the strain rate decreased. The hot workability of Ti6Al4V-5Cu alloy was also accessed from the viewpoint of the sub-grain structure.