目的:构建尼古丁诱导的小鼠生精细胞体外损伤模型,探讨小鼠骨髓间质干细胞来源的胞外囊泡(mouse bone marrow mesenchymal stem cell-derived extracellular vesicles,mBMMSCs-EVs)对于小鼠GC-1生精细胞损伤的修复作用.方法:采用不同浓度尼古丁(0、8、16、32μg/mL)处理GC-1生精细胞24、48、72 h,通过MTT实验筛选尼古丁诱导GC-1生精细胞损伤的最适浓度和时间;将GC-1生精细胞分为对照组、尼古丁组、尼古丁+胞外囊泡组;通过MTT实验、Transwell实验和流式细胞术凋亡实验观察mBMMSCs-EVs干预前后受损生精细胞GC-1生物学特性的变化.结果:MTT实验结果表明,32μg/mL尼古丁作用GC-1生精细胞24 h时,细胞增殖能力明显降低(P<0.05),mBMMSCs-EVs作用于尼古丁预处理后,细胞增殖能力明显增高(P<0.05);与对照组相比,尼古丁组GC-1生精细胞迁移能力明显降低,细胞凋亡能力明显升高(P均<0.01),与尼古丁组相比,尼古丁+胞外囊泡组生精细胞增殖和迁移能力明显升高,细胞凋亡能力明显下降(P<0.01或<0.05).结论:mBMMSCs-EVs可以逆转尼古丁引发的GC-1生精细胞增殖、迁移能力的损伤以及细胞凋亡.

目的:探讨miR-376c-3p在乳腺癌中的表达水平、生物学功能及分子调控机制.方法:通过实时荧光定量PCR检测乳腺上皮MCF-10A细胞及乳腺癌细胞中miR-376c-3p表达水平;然后,用miR-376c-3p mimic转染乳腺癌MCF-7与MDA-MB-231细胞,用平板克隆实验、划痕实验以及Transwell实验分别检测其增殖与迁移能力.利用Starbase数据库预测miR-376c-3p的下游靶基因,并通过蛋白质印迹实验进行验证.结果:与人乳腺上皮MCF-10A细胞相比,乳腺癌MCF-7和MDA-MB-231细胞中miR-376 c-3p表达水平显著降低(P＜0.01);平板克隆实验显示,过表达miR-376c-3p可以显著抑制乳腺癌细胞增殖(P均＜0.05);Transwell实验和划痕实验表明,miR-376c-3p表达上调可抑制乳腺癌细胞迁移能力;通过生物信息学预测分析,结果显示miR-376 c-3p可以与谷氨酸受体相互作用蛋白1(glutamate receptor interacting protein 1,GRIP1)结合,且蛋白质印迹实验证实过表达miR-376c-3p可以抑制乳腺癌细胞GRIP1表达,同时波形蛋白表达水平明显下调,E-钙黏蛋白表达明显上调.结论:在乳腺癌细胞中,miR-376c-3p表达下调;在体外,过表达miR-376c-3p可显著抑制乳腺癌细胞的增殖和迁移能力,可能通过靶向GRIP1调节乳腺癌细胞上皮间质转化进程.

目的:探讨人脐静脉内皮细胞来源胞外囊泡(human umbilical vein endothelial cell-derived extracellular vesi-cles,HUVEC-EVs)对乳腺癌细胞增殖和迁移的影响及其作用机制.方法:采用不同浓度(0、10、20、40μg/mL)HUVEC-EVs分别处理乳腺癌MDA-MB-231和MCF-7细胞24 h,通过MTT实验和平板克隆形成实验检测乳腺癌细胞增殖能力,Transwell实验检测乳腺癌细胞迁移和侵袭能力,划痕实验检测乳腺癌细胞迁移能力,蛋白质印迹实验检测乳腺癌细胞JAK2/STAT3信号通路相关蛋白的表达.结果:与0μg/mL HUVEC-EVs空白对照组相比,10、20、40μg/mL HUVEC-EVs处理组乳腺癌MDA-MB-231和MCF-7细胞增殖能力明显提高(P<0.05),平板克隆形成能力、迁移和侵袭能力也明显增强(P<0.05),总JAK2、STAT3蛋白无明显变化(P>0.05),而p-JAK2、p-STAT3蛋白表达升高(P<0.05).结论:HUVEC-EVs可能通过JAK2/STAT3信号通路的磷酸化促进乳腺癌细胞增殖及迁移.

目的:探讨6-磷酸果糖激酶-2/果糖-2,6-二磷酸酶同工酶3(6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases 3 isozyme,PFKFB3)在结直肠腺癌中的表达及其意义.方法:选取31例结直肠腺癌患者手术切除的癌组织及其对应的癌旁组织标本,采用实时荧光定量PCR(qRT-PCR)法检测PFKFB3 mRNA表达,免疫组织化学法检测PFKFB3蛋白的表达,并探讨免疫组化阳性表达率与临床病理参数间的关系.结果:qRT-PCR结果显示,31例结直肠腺癌组织PFKFB3基因的相对表达量为1.93±1.73,明显高于其相对应的癌旁组织,差异有统计学意义(t=-2.985,P<0.01).免疫组织化学结果显示,结直肠腺癌组织及其对应的癌旁组织PFKFB3阳性表达率分别为61.3%(19/31)和3.2%(1/31),差异有统计学意义(P<0.01).PFKFB3的高表达与肿瘤直径、分化程度、侵袭、转移及治疗前癌胚抗原水平有关(均P<0.05).结论:PFKFB3对结直肠腺癌具有潜在的临床诊断价值,且与肿瘤的恶性程度存在一定的相关关系.

Severe acute pancreatitis, one of the most common clinical acute abdominal diseases and one of the difficult diseases in clinical treatment, often happens in emergency with complex etiology and leads to high mortality. Multiple signaling pathways are involved in the pathogenesis and progression of severe acute pancreatitis, in which p38MAPK signal transduction pathway is the most notable. P38MAPK inhibitors, a kind of synthesis of small molecular organic compounds, by their specificity can suppress the expression and release of inflammatory mediators and can re-lieve the tissues injury. Thus, p38MAPK inhibitors are expected to become the effective way for treatment of severe acute pancreatitis.

Objective:To investigate the protective effects of clodronate-liposome on cerebral damage in-duced by severe acute pancreatitis(SAP)in rats.Methods:Clodronate-liposome was prepared by means of thin film.The models of SAP were performed by prinking injection of 5%sodium taurocholate (2 mL/kg) into the pancreatic capsule.A total of 48 male healthy SD rats were randomly divided into the sham group (Sham),SAP+liposome group (LB),SAP+clodronate-liposome group (LC).LB group were injected with liposome (4 mL/kg)while LC group were injected with clodronate-liposomes (4 mL/kg)in tail vein immediately after the SAP model performed.The change of amylase (AMS)in serum and the level of IL-6, TNF-αwere obtained at 2 h and 6 h after operation,respectively.Pathologic alteration of pancreas was ob-served and pathological score of pancreatic tissue was also estimated.The water and Evan′s blue content of brain tissues,the content of myelin basic protein (MBP)in cerebrospinal fluid were detected.Results:Compared with Sham group,the serum levels of AMS,IL-6,TNF-αand pancreatic tissue pathological score were significant elevated at 2 and 6 hours in LB group(P<0.01).In LC group,the serum levels of AMS, IL-6,TNF-αand pancreatic tissue pathological score were significant decreased at 2 and 6 hours compared with LB group(P<0.01).Compared with Sham group,the water and Evan′s blue content of brain tissues, the content of MBP in cerebrospinal fluid were significant elevated at 2 and 6 hours in LB group (P <0.01 ).In LC group,the water and Evan′s blue content of brain tissues,the content of MBP in cerebrospi-nal fluid were significant decreased at 2 and 6 hours compared with LB group(P<0.01 ).Conclusion:Monocytes/macrophages can be depleted by phagocytosis of liposome encapsulated clodronate,which may contribute to protection of clodronate-liposome against cerebral damage caused by SAP in rats.

目的:探讨氯膦酸二钠脂质体（LC）对大鼠重症急性胰腺炎（SAP）肺损伤的影响及与Akt、MAPK（ERK1/2）通路的关系。方法:将48只SD大鼠随机均分为假手术组、SAP模型组（模型组）、SAP模型+LC处理组（LC组）,后两组采用膜下注射5%牛磺胆酸钠制作SAP模型,并分别于造模后尾静脉注射空白脂质体与LC。各组分别于术后2、6h后检测血清淀粉酶（AMS）、IL-6、TNF-α的含量,观察肺组织病理学变化,及肺组织中Akt和MAPK（ERK1/2）的表达。结果:与假手术组比较,模型组与LC组血清AMS、IL-6及TNF-α含量、肺组织病理学评分,肺组织Akt和MAPK（ERK1/2）表达水平均明显升高,且均随时间延长而更加明显（均P＜0.05）,但LC组的上述指标在各时间点上均明显低于模型组（均P＜0.05）。结论:LC有减轻大鼠SAP肺损伤的作用,机制可能与肺泡巨噬细胞吞噬LC后,Akt和MAPK（ERK1/2）信号通路抑制,从而减少炎症细胞因子的释放有关。