Evaluation of paramalleolar and inframalleolar bypasses in dialysis- and nondialysis-dependent patients with critical limb ischemia

Objective

The aim of this study was to elucidate the efficacy of paramalleolar or inframalleolar bypass (PIMB) in hemodialysis-dependent (HD) patients with critical limb ischemia (CLI) and nonhemodialysis-dependent (NHD) patients in terms of clinical outcomes.

骨髓基质细胞联合细胞因子对脐血CD133+细胞体外扩增作用的研究

为了探讨胚胎骨髓基质细胞（FBMSC）联合细胞因子对脐血单个核细胞（MNC）中CD133＋细胞的体外扩增作用,将新鲜脐血（CB）中分离出来的MNC接种于无血清培养体系中培养14天。实验分为4组：C组为空白对照组,不含基质细胞和细胞因子;S组为单用基质细胞组;F组为单用细胞因子组;SF组为联合使用基质细胞和细胞因子组。在第0,6,10及14天检测有核细胞总数、CD133＋细胞数及集落形成单位（CFU）数。结果表明：各时间点SF组有核细胞总数的扩增倍数均高于其它组;除了第14天外,SF组在第6、10天时CD133＋细胞数、CFU数的扩增倍数均高于其它组。结论：胚胎骨髓基质细胞对延缓造血细胞的分化具有重要的作用,基质细胞联合细胞因子可以有效的扩增脐血单个核细胞及其中的CD133＋细胞,这是一种比较接近于临床移植要求的造血细胞体外扩增方法。     

Up-regulation of miR-181a in clear cell renal cell carcinoma is associated with lower KLF6 expression,enhanced cell proliferation,accelerated cell cycle transition,and diminished apoptosis

Objectives

Dysregulated expression of miR-181a accompanies tumorigenesis in many human cancers. However, in clear cell renal cell carcinoma (ccRCC), the role of miR-181a remains unclear. The aim of this study was to investigate biological functions of miR-181a and its expression levels in ccRCC tissues and cancer cell lines.

ADAM12基因表达沉默对CD133阳性胶质瘤细胞自我更新能力的影响

目的研究解整合素-金属蛋白酶12(a disintegrin and metalloprotease 12,ADAM12)基因表达沉默抑制CD133阳性(CD133+)胶质瘤细胞的自我更新能力。方法采用sh RNA重组慢病毒转染技术沉默胶质瘤U87细胞系ADAM12基因表达分为ADAM12基因表达干扰序列组(sh RNA-ADAM12)、阴性对照组(sh RNA-NC)及空白对照(sh RNA-C)组。通过Western blotting以及Real-time PCR验证各组细胞ADAM12表达情况;采用悬浮培养得到胶质瘤细胞球以富集CD133+的胶质瘤细胞;并通过免疫荧光染色技术检测ADAM12与CD133在细胞球与贴壁细胞的表达情况;通过神经肿瘤球形成实验检测三组细胞的自我更新能力;利用Western blotting分别检测三组细胞成球后未分化或分化相关蛋白CD133、GFAP及TUBB3以及Notch通路靶基因Hes1的蛋白表达情况。结果慢病毒转染技术可显著下调U87胶质瘤细胞ADAM12的m RNA及蛋白的表达量,sh RNA-ADAM12组与sh RNA-NC组较sh RNA-C组m RNA的相对表达量为0.22±0.03与0.98±0.06(F=425.37,P0.01);三组ADAM12蛋白的相对表达量分别为28.72%±2.36%、69.21%±3.92%及69.04%±3.57%(F=145.42,P0.01);免疫荧光染色显示细胞球中ADAM12与CD133表达量明显高于普通细胞;神经肿瘤球形成实验结果显示,三组成球数分别为45.5±2.3、104.2±5.8以及109.6±6.2,与sh RNA-NC组及sh RNA-C组相比,sh RNA-ADAM12组的成球能力明显降低,差异具有统计学意义(F=147.03,P0.01)。sh RNA-ADAM12组与sh RNA-C组相比,GFAP与TUBB3蛋白表达量分别上调约166%与146%,CD133与HES1的蛋白表达量分别下调了54%与50%,差异均具有统计学意义(P0.01)。结论 ADAM12基因表达沉默可能通过抑制Notch通路活性降低CD133+胶质瘤细胞的自我更新能力。     

Silencing LINC01234 represses pancreatic cancer progression by inhibiting the malignant phenotypes of pancreatic cancer cells

ObjectivePrevious works have outlined the pivotal involvement of long intergenic non-coding RNA (lincRNA) in cancer progression, while the efficiency of LINC01234 in pancreatic cancer remained obscure. The purpose of this research is to unravel the regulatory mechanism of LINC01234 in pancreatic cancer via modulating microRNA (miR)-513a-3p and hexose 6-phosphate dehydrogenase (H6PD).MethodsPancreatic cancer cells were cultured and clinical tissue specimens were collected. LINC01234, miR-513a-3p and H6PD levels in pancreatic cancer cells and tissues were examined. Plasmids altering LINC01234, miR-513a-3p and H6PD expression were transfected into pancreatic cancer cells to assess the change in biological behaviors of pancreatic cancer cells. The targeting relations among LINC01234, miR-513a-3p and H6PD were validated.ResultsLINC01234 and H6PD levels were elevated while miR-513a-3p level was reduced in pancreatic cancer cells and tissues. LINC01234 deficiency hindered the malignant biological activities of pancreatic cancer cells. MiR-513a-3p depletion or H6PD elevation could abrogate the inhibitory effects of LINC01234 silencing on pancreatic cancer cells. LINC01234 sponged miR-513a-3p that targeted H6PD.ConclusionThe reduced LINC01234 exerts inhibitory impacts on pancreatic cancer cells via targeting miR-513a-3p to restrain H6PD level. The current study broadens the understanding of LINC01234 function and affords novel therapeutic targets for pancreatic cancer treatment.     

Fibroblast-derived exosomal miRNA-133 promotes cardiomyocyte-like differentiation

ObjectiveTo investigate the role of exosomal miRNA-133 secreted by cardiac fibroblasts (CFs) in promoting cardiomyocyte differentiation.MethodsNeonatal rat CFs were cultured in vitro, and the cultured CFs were divided into three groups as follows: induction, miRNA-133 high expression, and miRNA-133 inhibition. miRNA-133 was transfected into CFs with lentivirus as a vector. CFs were transfected with the miRNA-133 inhibitor, and the markers of cardiomyocyte were detected through immunofluorescence staining, Western blotting, and real-time quantitative polymerase chain reaction (qRT-PCR) at 3, 8, and 14 days, respectively. The expression levels of cardiac troponin T (cTnT) and cardiac actin (α-actin) were determined, and qRT-PCR was used to detect the expression of miRNA-133 in the fibroblast exosomes.ResultsCFs subjected to immunofluorescence staining expressed vimentin and discoid domain receptor 2. The exosomes secreted by CFs were observed as small vesicles of 30–100 nm via transmission electron microscopy, and Western blotting was used to detect exosome-specific protein CD63 and CD9 expression. The expression levels of cTnT, α-actin, and exosomal miRNA-133 secreted into the supernatant of the miRNA-133 high-expression group increased gradually at different time points and reached the highest level at 14 days. The expression levels of cTnT, α-actin, and exosome miRNA-133 in the miRNA-133 inhibition group were the lowest.ConclusionThe exosomal miRNA-133, which is derived from CFs, can promote the differentiation of fibroblasts into cardiomyocyte-like cells.     

抑制miR-133b通过靶向FOXP3对PD大鼠调节性T淋巴细胞、炎性反应和神经元凋亡的影响

目的探究抑制微小RNA(microRNA,miR)-133b通过靶向叉头盒蛋白3(forkhead box protein 3,FOXP3)对帕金森病(Parkinson’s disease,PD)大鼠调节性T细胞(regulatory T cells,Treg)的影响。方法 32只PD模型大鼠随机分为PD组和PD+miR-133b antagomir组(n=16),健康大鼠16只作为对照组。尾静脉注射miR-133b antagomir(300μg)来抑制miR-133b的水平。检测和比较各组大鼠神经功能、黑质损伤、细胞凋亡、炎性反应、Treg细胞水平、miR-133b、FOXP3 mRNA和蛋白表达水平;通过双荧光素酶报告验证miR-133b和FOXP3的靶向关系。结果 PD组的逃避潜伏期、旋转速率、细胞凋亡情况、IL-6、miR-133b水平显著高于对照组,穿越次数、IL-10、FOXP3 mRNA和蛋白表达量显著低于对照组(P<0.05)。PD+miR-133b antagomir组的逃避潜伏期、旋转速率、细胞凋亡情况、IL-6、miR-133b水平显著低于PD组,穿越次...     

Expression of AC133 and CD117 on candidate normal stem cell populations in childhood B-cell precursor acute lymphoblastic leukaemia

To identify residual candidate normal progenitor/stem cell populations in childhood B-cell precursor acute lymphoblastic leukaemia (ALL), expression of AC133 and CD117 was analysed on the leukaemic cell clone and on immature B-lineage-negative CD34+CD19- bone marrow cells. 10/25 patients (40%) had no detectable expression of AC133 within the leukaemic cell clone. 24/26 patients (92%) lacked expression of CD117 on the leukaemic blast cell population. In contrast, a distinct AC133-positive cell population was found in 8/8 children with AC133-negative ALL and a CD117-positive cell population could be identified in 12/12 children with CD117-negative ALL, within the CD34+CD19- progenitor/stem cell compartment. These observations provide further evidence that in B-cell precursor ALL, unlike in acute myelogenous leukaemia, it may be possible to distinguish residual normal progenitor/stem cells from the leukaemic cell clone.     

miR-451对胶质瘤细胞株A172生物学特征的影响

目的 研究miR-451对胶质瘤细胞株A172生物学特征的影响.方法 合成寡核苷酸miR-451拟似物(miR-451 mimics)转染胶质瘤细胞以上调miR-451的表达,real-time PCR检测转染后miR-451的表达,Western印迹法检测目标蛋白表达,应用流式细胞术、MTT法评价细胞生长和增殖的生物学特征变化.结果 转染miR-451 mimics后,real-time PCR检测提示miR-451表达升高,Western blot结果显示癌基因C-myc表达下降＞63.6%,细胞周期正向调节因子CDK2、CDK4、Cyclin D1和Cyclin E表达下降;细胞周期分析表明miR-451 mimics组进入G0/G1期的细胞数较对照组增多达17.4%,出现G0/G1期阻滞;MTT法分析显示miR-451 mimics组细胞生长受抑.结论 miR-451可以抑制人脑胶质瘤细胞生长和增殖能力,具有抑瘤作用.     

肺腺癌A549细胞CD133~+CD44~+表型在裸鼠体内成瘤能力的研究

目的 探讨肿瘤标志物CD133和CD44的表达与人肺腺癌A549细胞株在裸鼠体内形成肿瘤能力的关系.方法 使用单细胞克隆的方法 ,筛选出能够持续增殖分裂的肿瘤细胞,应用免疫荧光方法 和流式细胞仪检测CD133和CD44在具有增殖分裂能力的A549细胞和普通A549肿瘤细胞的表达情况;按表达结果 对A549细胞进行分组,应用单克隆方法 对各组细胞进行培养,并按不间密度注射于裸鼠皮下,观察各个组别的成瘤能力.结果 (1)在倒置显微镜下观察肿瘤细胞生长情况,发现多数细胞死亡,1周后仅有4.11%的细胞能够增殖分裂形成克隆.(2)具有增殖分裂能力肿瘤细胞的免疫荧光结果 :CD133阳性率为19.0%,CD44阳性率为57.0%;普通A549肿瘤细胞免疫荧光结果 :CD133阳性率为2.5%,CD44阳性率为34.0%;具有分裂增殖能力的肿瘤细胞CD133和CD44阳性表达率显著高于普通A549肿瘤细胞(P<0.01).(3)裸鼠体内成瘤能力实验提示:CD133~+CD44~+(A组)细胞成瘤能力显著高于CD133~-CD44~-(B组)、CD133~-CD44~+(C组)和CD133~+CD44~-(D组)细胞(P<0.05);裸鼠瘤块病理检查证实均为腺癌,各脏器检查未发现转移:在接种CD133~+CD44~+(A组)细胞的瘤块组织中发现除了有CD133~+CD44~+细胞外,还有CD133~-CD44~-细胞.结论 肺腺癌A549细胞株中CD133~+CD44~+细胞在裸鼠体内的成瘤能力显著高于其他细胞.