miR-146a-5p regulates autophagy and NLRP3 inflammasome activation in epithelial barrier damage in the in vitro cell model of ulcerative colitis through the RNF8/Notch1/mTORC1 pathway

Ulcerative colitis (UC) is a chronic inflammatory disease affecting the colon that can be influenced by microRNAs (miRNAs). This study aims to investigate the impact of miR-146a-5p on lipopolysaccharide (LPS)-induced Caco-2/HT-29 cell autophagy and NLRP3 inflammasome activation and the underlying mechanism, with the aim of identifying potential therapeutic targets. We used LPS to establish Caco-2/HT-29 cell models and measured cell viability by CCK-8. The levels of miR-146a-5p, RNF8, markers of NLRP3 inflammasome activation and autophagy, proteins involved in the Notch1/mTORC1 pathway, and inflammatory factors were assessed by RT-qPCR, Western blot, and ELISA. Intestinal epithelial barrier function was evaluated by measuring transepithelial electrical resistance. Autophagic flux was measured using tandem fluorescent-labeled LC3. miR-146a-5p was highly-expressed in LPS-induced Caco-2/HT-29 cells, and autophagy flux was blocked at the autolysosomal stage after LPS induction. Inhibition of miR-146a-5p suppressed NLRP3 inflammasome activation, reduced intestinal epithelial barrier damage, and facilitated autophagy inhibition in LPS-induced Caco-2/HT-29 cells. The autophagy inhibitor NH4Cl partially nullified the inhibitory effects of miR-146a-5p inhibition on NLRP3 inflammation activation. miR-146a-5p targeted RNF8, and silencing RNF8 partly abrogated the action of miR-146a-5p inhibition on promoting autophagy and inhibiting NLRP3 inflammasome activation. miR-146a-5p inhibition suppressed the Notch1/mTORC1 pathway activation by upregulating RNF8. Inhibition of the Notch1/mTORC1 pathway partially nullified the function of silencing RNF8 on inhibiting autophagy and bolstering NLRP3 inflammasome activation. In conclusion, miR-146a-5p inhibition may be a potential therapeutic approach for UC, as it facilitates autophagy of LPS-stimulated Caco-2/HT-29 cells, inhibits NLRP3 inflammasome activation, and reduces intestinal epithelial barrier damage by upregulating RNF8 and suppressing the Notch1/mTORC1 pathway.     

疏肝理脾片抗大鼠免疫性肝纤维化研究

用牛血清白蛋白（ＢＳＡ）造成大鼠免疫损伤性肝纤维化模型，观察疏肝理脾片对肝纤维组织增生程度、肝羟脯氨酸（Ｈｙｐ）含量和血清透明质酸（ＨＡ）及肝功能的影响。结果表明疏肝理脾片能明显降低肝Ｈｙｐ含量，减轻肝纤维组织增生程度，并较好地提高血清白蛋白水平     

烧烫安乳膏对大鼠皮肤深二度烫伤创面愈合的影响

目的探讨烧烫安乳膏对大鼠深二度烫伤的影响。方法采用大鼠深二度烫伤模型,创面分别外用高、中、低剂量的烧烫安乳膏,通过伤后不同时间点创面取材,检测烫伤创面羟脯氨酸水平、创面愈合面积百分比、愈合时间及观察组织病理学变化,以基质乳膏作阴性对照,湿润烧伤膏作阳性对照,观察烧烫安乳膏对大鼠深二度烫伤创面愈合的影响。结果创面外用中、高剂量的烧烫安乳膏可以增加创面羟脯氨酸水平,促进肉芽组织生长,促进创面局部成纤维细胞生长因子-1β(FGF-1β)的表达,缩短愈合时间;作用与湿润烧伤膏相当。结论烧烫安乳膏可增强胶原合成,促进上皮细胞增殖,加速烫伤创面愈合。     

吡格列酮对炎性因子所致胰岛细胞凋亡的保护作用

目的：探讨Ppar-γ激动剂吡格列酮（PGZ）对炎性因子所诱导的胰岛细胞凋亡的保护作用，初步研究其作用机制。方法：Western—blotting检测NIT-1细胞的Ppar-γ受体蛋白表达；联合IL-1β及IFN-γ作用于体外培养的胰岛细胞株NIT-1，建立胰岛细胞炎症模型，观察（IL-1β＋IFN-γ）与PGZ预孵育NIT-1细胞的凋亡；MTT法、流式细胞术检测细胞生长抑制率及凋亡率，比色法检测凋亡细胞caspase-3比活性。结果：NIT-1细胞表达Ppat-γ蛋白；（IL—1β＋IFN-γ）组作用24h对NIT-1细胞生长抑制率、凋亡率、caspase-3比活性分别为49．8％、28．0％、3．5，对照组3项指标分别为0％、4．3％、1，两组比较均有十分显著差异（P〈0．01）；PGZ组3项指标分别为2．7％、7．1％、1.3，与对照组比较，均无明显差异（P〉0．05）；（PGZ＋IL-1β＋IFN-γ）组的3项指标分别为18．6％、14．8％、1．5，与（IL-1β＋IFN-γ）组比较均有差异（P〈0．05），与对照组比较分别为P〉0．05，P〈0．05，P〈0．05。结论：NIT-1细胞表达Ppar-γ受体蛋白；联合IL—1β及IFN-γ明显诱导NIT-1细胞凋亡，PGZ显著抑制IL-1β及IFN-γ诱导的NIT-1细胞凋亡，其机制与降低caspase-3活性有关。     

中医医院膝关节骨关节炎住院患者的治疗情况--附892份病历分析

为了解中医医院膝关节骨关节炎住院患者的治疗现状,对湖南省12家中医医院5年膝关节骨关节炎住院患者治疗情况进行调查,摘录其相关治疗资料,并进行统计学分析处理.结果显示892例中,78.8%者内服中药汤剂,19.4%外用中药,所用中药性味多为辛温,内服药以祛风湿、补虚类药物居多,外用药以祛风湿、活血化瘀类药物为主.25.1%采用针灸治疗,常用穴位有膝眼、足三里、阳陵泉、阴陵泉、血海、梁丘等.13.1%接受推拿按摩治疗,但大多数病案未记载具体手法.46.2%内服NSAIDs,19.5%使用抗生素,15.4%使用激素类药物.表明中医医院对膝关节骨关节炎住院患者常采用中西医结合方法治疗,中医治疗多为各种方法组合的综合疗法,中药多选择性味辛温之品,祛风湿药、补虚药和活血化瘀类药物为临床常用中药.     

基于影像组学分类器术前预测直肠非黏液性腺癌淋巴结转移

目的：探讨影像组学方法在术前预测直肠非黏液性腺癌淋巴结转移中的价值。方法：回顾性分析91例手术病理切片证实为直肠非黏液性腺癌患者的影像学资料,其中61例为训练样本,30例为验证样本。基于全瘤体积,从每个原发病灶术前高分辨T2加权成像(T2-weighted imaging,T2WI)图像中提取影像组学特征1 301个。基于训练样本,利用最小绝对收缩和选择算子(the least absolute shrinkage and selection operator,LASSO)逻辑回归方法筛选关键特征并构建影像组学分类器。采用受试者工作特征(receiver operating characteristic,ROC)曲线评价影像组学分类器的辨别效能,并将其与形态学标准进行比较。在验证样本中验证影像组学分类器的价值。结果：由5个影像组学特征构建的分类器与淋巴结转移状态有关(P<0.001)。在训练样本和验证样本中,影像组学分类器诊断淋巴结转移的曲线下面积分别为0.874(95% CI：0.787~0.960)和0.878(95% CI：0.727~1.000),形态学标准诊断淋巴结转移的曲线下面积分别为0.619(95% CI：0.487~0.752)和0.556(95% CI：0.355~0.756)。无论是训练样本还是验证样本,影像组学分类器的诊断效能均高于形态学标准(均P<0.05)。结论：影像组学分类器可术前个体化预测直肠非黏液性腺癌淋巴结转移,而且其诊断效能高于形态学标准。     

HIF1α-siRNA and gemcitabine combination-based GE-11 peptide antibody-targeted nanomedicine for enhanced therapeutic efficacy in pancreatic cancers

Abstract

Pancreatic cancer is one of the deadliest cancers across the world with an average 5-year survival rate of less than <6%. In this study, gemcitabine (GEM) and HIF1α-siRNA loaded GE-11 peptide conjugated liposome was successfully prepared and evaluated for its antitumor efficacy in pancreatic cancer cells. The GE11 increased the targeting specificity of liposome carrier and increased the intracellular concentrations in the cancer cells. Furthermore, synergistic combination of GEM and HIF1a-siRNA exhibited remarkable improvement in the declining of cancer cell proliferations. siRNA could effectively decrease the expression of HIF1a gene in the cancer cells. Importantly, GE-11 peptide-conjugated GEM/siRNA-loaded liposomes (GE-GML/siRNA) increased the total amount of apoptosis cells with higher proportion of cells in late apoptosis phase. GE-GML induced remarkable apoptosis of cancer cells and induced chromatin condensation and nuclear fragmentation which are considered to be typical features of apoptosis and cell death. GE-GML/siRNA showed a significant reduction in the tumour burden suggesting the superior anticancer efficacy of this formulation. GE-GML/siRNA showed four-fold reduction in tumour compared to control and two-fold reduction compared to GE-GML, respectively. Overall, present work lays foundation for the combination of GEM and HIF1a-siRNA loaded in a targeted nanocarrier system as a unique therapeutic option in pancreatic cancer treatment.     

KA1表达再分布对神经元兴奋毒性的影响

背景:特异的兴奋性神经递质受体过度活化时便发生细胞的兴奋性中毒,有关NMDA受体和AMPA受体活化致细胞兴奋性中毒死亡通路较为明确,但关于红藻氨酸(Kainate,KA)受体的功能到目前为止仍未明了。目的:探究小鼠海马内注射KA之后KA1受体的再分布状况对中枢神经元兴奋性中毒的影响。方法:成年雄性C57BL6小鼠海马内注射KA或PBS,2h后进行Bederson体征评分、全脑切片免疫组化分析和死亡神经元检测。结果与结论:海马内注射KA2h后,Bederson体征评分表明中枢神经功能出现明显损伤;KA1受体表达的变化主要出现在海马CA1和CA3区,在CA1区的表达上调明显,并开始出现神经元死亡,4~6h后可见大量神经元死亡。结果表明KA促使KA1受体亚单位在海马神经元CA区重新分布,增加神经元对兴奋性氨基酸毒性的敏感性,导致神经细胞死亡,中枢神经功能缺失。     

葡萄糖转运蛋白1和波形蛋白在肺癌胸水中的应用价值

目的:采用免疫细胞化学技术测定葡萄糖转运蛋白1(GLUT1)及波形蛋白(VM)在癌细胞中的定位表达,探讨GLUT1和VM在肺癌胸水中的应用价值。方法:以肺癌患者为研究对象,以肺癌性胸水为检测基础,收集胸水沉淀细胞,制成石蜡切块,采用免疫组化技术-链霉菌抗生物素-过氧化物霉法检测30例肺癌胸水和20例良性胸水中GLUT1和VM蛋白表达,统计单个及联合应用价值。结果:30例肺癌胸水中,GLUT1表达阳性的有25例(86.7%),良性胸水中间皮细胞VM表达阳性的有18例(90.0%),两种不同性质的胸水GLUT1和VM的阳性率表达均有差异(P<0.05),GLUT1和VM在肺癌胸水中表达呈负相关(P<0.05),两者联合诊断灵敏度为90%。结论:应用免疫组化方法检测肺癌胸水中GLUT1和VM蛋白表达,可提高肺癌早期诊断的灵敏度。