目的 分析促红细胞生成素（EPO）及促红细胞生成素受体（EPOR）在肝细胞癌（NCC）以及正常组织中的表达规律,以及它们和肝细胞癌微血管密度（MVD）之间的关系。方法 选取我院手术切除的肝细胞肝癌的标本30例,取肿瘤边缘2.0 cm的肝组织作为对照,同时取正常肝脏组织10例做为阴性对照。利用酶联免疫吸附实验（ELISA）检测各组织中EPO及EPOR表达水平,利用免疫组织化学方法染色检测微血管密度（MVD）。对比癌组织和癌旁组织EPO、EPOR及MVD差异,分析NCC中EPO、EPOR、MVD与肿瘤病理特征的关系,分析EPO、EPOR表达水平与MVD之间的关系。结果 HCC组织中,EPR、EPOR、MVD均高于癌旁组织和正常组织,差异有统计学意义（P<0.001）,EPR、EPOR、MVD在癌旁组织和正常组织中,差异无统计学意义（P>0.05）。肿瘤大小>5 cm、存在包膜侵犯、存在远处转移以及高中分化的HCC中,EPR、EPOR、MVD水平高于肿瘤大小≤5 cm、无包膜侵犯、无远处转移以及低分化的水平,差异有统计学意义（P<0.05）。Person相关分析结果显示,EPO表达水平与MVD的相关系数r=0.651（P<0.001）,EPOR表达水平与MVD的相关系数r=0.620（P<0.001）。结论 EPO、EPOR、MVD在HCC中呈现高水平,且与肿瘤大小、局部侵犯、远处转移及分化程度有关,其机制可能与EPO、EPOR增加MVD有关。

Objective To analyze the expression of erythropoietin （EPO） and erythropoietin receptor （EPOR） in hepatocellular carcinoma （NCC） and normal tissues,and their relationship with hepatocyte microvessel density （MVD）. Methods Thirty specimens of hepatocellular carcinoma hepatectomy were selected from our hospital. The liver tissue at the edge of the tumor was taken as a control,and 10 cases of normal liver tissue were used as a negative control. The expression levels of EPO and EPOR in each tissues were detected by enzyme-linked immunosorbent assay （ELISA）,and microvessel density （MVD） was detected by immunohistochemistry. The differences of EPO,EPOR and MVD between each tissues were compared. The relationship between EPO,EPOR,MVD and tumor pathological features in NCC was analyzed. The relationship between EPO and EPOR expression levels and MVD was analyzed. Results In HCC tissues,EPR,EPOR and MVD were higher than those in adjacent tissues and normal tissues. The difference was statistical difference （P<0.001）. EPR,EPOR and MVD were not statistically significant in adjacent tissues and normal tissues. P>0.05）. The levels of EPR,EPOR,and MVD in tumors with tumor size >5 cm,invasion of the capsule,distant metastasis,and high-differentiation were higher than those of tumor size ≤ 5 cm,no capsule invasion,no distant metastasis,and poor differentiation. The difference was statistical difference （P < 0.05）. Person correlation analysis showed that the correlation coefficient between EPO expression level and MVD was r=0.651 （P<0.001）,and the correlation coefficient between EPOR expression level and MVD was r=0.620 （P<0.001）. Conclusion EPO,EPOR and MVD are highly expressed in HCC,and are related to tumor size,local invasion,distant metastasis and differentiation. The mechanism may be related to EPO and EPOR increasing MVD.

目的 筛选结直肠癌（CRC）差异性表达的长链非编码RNA（lncRNA）,并进行临床标本验证,研究其对结肠癌细胞HCT116功能的作用。方法 利用lncRNA PCR芯片对3对CRC组织和癌旁对照组织筛选差异性表达的lncRNA,确定候选研究lncRNA GAPLINC,RT-qPCR对21例临床样本进行验证其表达的差异性;同时构建GAPLINC表达质粒及其沉默体siRNA转染HCT116细胞,研究其对细胞凋亡、迁移及侵袭能力的影响。结果 lncRNA芯片实验结果提示CRC组织中存在大量的差异性表达的lncRNA,其中GAPLINC在CRC组织表达稳定增加,21例临床样本进一步验证了其在肿瘤组织中表达增加（P<0.05）;转染GAPLINC表达质粒后,HCT116细胞凋亡被抑制,同时其迁移及侵袭能力增强,转染siRNA抑制GAPLINC的表达后,则出现相反的结果。结论 利用lncRNA芯片可对CRC差异性表达lncRNA进行批量筛选,GAPLINC在CRC组织中表达稳定增加,具有促癌作用,在CRC发生发展中可能起着重要作用。

Objective To screen the differentiational expression of lncRNA in CRC tissue,confirm it in large simple of clinical specimens,and study its effects on human colorectal cells HCT116 cell line. Methods We screened the lncRNA which expressed differently in 3 CRC tissues and their pair-non carcinour tissues by lncRNA arrays;chose the over expressed lncRNA which played the potential role of oncogene for further researching,and tested the difference in 21 clinical specimens by RT-qPCR. We cultured the HCT166 cells,and then constructed expressed plasmids pcCDNA3.1-GAPLINC and synthesized GAPLINC siRNA,transfected the plasmids and siRNA into HCT116 cells;to study the changes of HCT116 cells behavior,the transwell assays were carried on;the changes of apoptosis of HCT116 cells were tested by flow cytometry. Results There existed many lncRNA which expressed differently between CRC tissues and normal control tissues by lncRNA arrays,there were 21 lncRNA down expressed,and 3 lncRNA up expressed;among these lncRNA,GAPLINC over expressed stably,and its high level of expression was approved in 21 clinical specimens by the test of RT-qPCR. We constructed the expressed plasmids pcCDNA3.1-GAPLINC and synthesizing GAPLINC siRNA successfully;after transfecting pcCDNA3.1-GAPLINC into HCT116 cells,the over expression of GAPLINC increased the migration and invasion of the HCT166 cells （P<0.05）,decreased the proportion of cell apoptosis （P <0.05）;by contraries,knocked down the expression of GAPLINC inhibited invasion and migration of HCT116 cells （P<0.05）,and promoted the apoptosis of the HCT116 cells （P <0.05）. Conclusion It could screen the different expression of lncRNA in large quantities by lncRNA arrays,and GAPLINC expressed highly and stably in CRC tissues. GAPLINC played a role of oncogene,which promoted the proliferation and invasion of CRC cells,and inhibited the apoptosis of CRC cells,which meant playing an important role in the carcinoma and development of CRC.

目的 探讨血清ApoA-1浓度与食管鳞状上皮细胞癌的相关性及其预后价值。方法 对我院2010年1月—2012年10月收治的食管鳞状上皮细胞癌患者的血清ApoA-1浓度进行回顾性调查分析,与我院体检中心2009年1月—2009年4月219例正常人群的血清ApoA-1浓度作为对照组进行统计学处理分析。结果 210例食管鳞状上皮细胞癌患者组和219例健康对照组在性别和年龄比较均无统计学差异(P>0.05)。血清ApoA-1 浓度在ESCC组的pTNM和临床分期均有统计学差异(P<0.05)。与ApoA-1水平高的患者相比,ApoA-1水平低的患者的总体生存率更低(P=0.002)。结论 血清ApoA-1 水平是食管鳞状上皮细胞癌食管癌的一个重要的预后因素。

目的 采用生物信息学方法预测低氧预处理人胎盘绒毛膜间充质干细胞环状RNAs相对应的miRNA及其靶基因,并分析靶基因所参与的生物学过程和信号通路。方法 用Arraystar公司的商业软件为环状RNAs预测其相对应的miRNAs,分别用targetScan7.1和mirdbV5数据库预测miRNAs的靶基因,并取两个预测结果的合集,应用在线网站http://www.geneontology.org和http://www.genome.ad.jp/kegg对靶基因进行功能富集分析和信号通路富集分析。结果 功能富集分析表明,circRNAs的靶基因主要涉及到细胞发育、细胞分化和细胞发育调控。东京基因和基因组百科全书信号通路富集分析表明肿瘤中转录失控和有丝分裂原激活蛋白激酶(MAPK)信号通路最有意义,而且分析发现MAPK信号通路为核心通路。本研究表明,低氧预处理使得间充质干细胞中部分circRNAs的表达量发生差异性变化。结论 低氧预处理人胎盘绒毛膜间充质干细胞环状RNAs同低氧预处理间充质干细胞的生物学特性变化密切有关,为了解低氧预处理影响间充质干细胞特性发生变化的分子机制提供新思路。

Objective To predict the miRNA and its target genes of circular RNAs in hypoxia- preconditioned human palcenta chorionic mesenchymal stem cells using bioinformatics, and analyze the biological process and signaling pathway. Methods Arraystar's commercial software was used to predict the corresponding miRNAs of circular RNAs. The target genes of miRNAs were predicted by targetScan7.1 and mirdbV5 databases respectively, and an intersection of two prediction results was obtained. The online databases http://www. geneontology.org and http://www.genome.ad.jp/kegg performed functional enrichment analysis and signal pathway enrichment analysis of target genes. Results Functional enrichment analysis indicated that the target genes of circRNAs mainly involved cell development, cell differentiation and cell development regulation. The signal enrichment analysis of the Tokyo Gene and Genome Encyclopedia indicates that transcriptional misregulation in cancer and mitogen-activated protein kinase (MAPK) signaling pathway are most meaningful, and the MAPK signaling pathway is found to be the core pathway. This study showed that hypoxic preconditioning caused significant changes in the expression of mesenchymal stem cell circRNAs. Conclusion The changes of circular RNAs in hypoxia-preconditioned human placental chorionic mesenchymal stem cell is closely related to the biological characteristics of hypoxia-preconditioned mesenchymal stem cells. This study provides a new idea for understanding the molecular mechanism of hypoxic preconditioning affecting the changes of biological characteristics in mesenchymal stem cells.

目的 本研究以ICSI后未成熟卵母细胞为研究对象,分析比较卵母细胞不同发育阶段冷冻对其后续效果的影响,评估ICSI后未成熟卵母细胞的利用价值。方法 未成熟卵母细胞直接成熟培养(新鲜组)与玻璃化冷冻后成熟培养(冷冻组)的成熟率,并利用孤雌激活的方法比较卵母细胞的发育潜力。结果 发现新鲜组与冷冻组体外培养卵母细胞成熟率、受精率、卵裂率、优质胚胎率和囊胚率均无差异(P>0.05)。但两组的GV期卵母细胞成熟率低于MI期(P<0.05),且冷冻组的GV期卵母细胞受精率低于MI期(P<0.05)。不过裸卵体外成熟培养效果欠佳,特别是对GV期卵母细胞,体外成熟培养后的卵母细胞发育潜力低下,无囊胚形成。结论 ICSI后未成熟卵母细胞的冷冻对卵母细胞的发育潜力没有明显影响,但体外成熟培养的卵母细胞发育潜力低下,有待进一步提高体外成熟培养技术。

Objective In this study, the immature oocytes after ICSI were used to analyze the effects of freezing on the subsequent development of oocytes at different developmental stages, and to evaluate the utilization value of immature oocytes after ICSI(intracytoplasmic sperm injection). Methods The immature oocytes was directly cultured (fresh group) and matured after vitrification (frozen group), and the development potential of the oocytes was compared by parthenogenetic activation. Results There was no significant difference in the oocyte maturation rate, fertilization rate, embryo cleavage rate, high quality embryo rate and blastocyst rate between the fresh and frozen groups (P>0.05). However, the maturation rate of GV oocytes in the two groups was lower than MI oocytes (P<0.05), and the fertilization rate of GV oocytes in the frozen group was lower than MI oocytes (P<0.05). However, the in vitro maturation of naked oocytes was not effective, especially for GV oocytes, the oocyte development potential after in vitro maturation was low, there was no blastocyst formation. Conclusion The freezing of immature oocytes after ICSI has no significant effect on the development potential, but the development potential of naked oocytes matured in vitro was low, and the in vitro maturation culture technology of naked oocyte needs to be further improved.

目的 探讨红细胞分布宽度(RDW)与稳定性冠心病的冠脉病变严重程度的关系,评价RDW对稳定性冠心病的预测价值。方法 收集我院2016年11月—2018年11月期间因胸痛住院的患者330例,根据冠脉造影确诊稳定性冠心病组200例,除外冠心病130例为对照组。比较2组RDW水平的差异,分析RDW与冠脉病变严重程度(Gensini评分)的相关性及稳定性冠心病的独立危险因素。结果 稳定性冠心病组RDW水平高于对照组(P<0.05),稳定性冠心病组RDW与 Gensini评分之间存在正相关(r=0.217,P=0.002);多因素logistic回归分析显示RDW(OR=2.950,95%CI: 1.986~4.381,P<0.001)是稳定性冠心病的独立危险因子。结论 RDW与稳定性冠心病的冠脉病变严重程度呈正相关,RDW是稳定性冠心病的独立危险因素,为稳定性冠心病的诊断提供一定预测价值。

Objective To investigate the relationship between the distribution width of red blood cells (RDW) and the severity of coronary artery disease in stable coronary artery disease(SCAD), and to evaluate the predictive value of RDW for SCAD. Methods Patients who were hospitalized for chest pain from November 2016 to November 2018 were enrolled (n=330). According to coronary angiography,200 patients were divided into SCAD group (n=200),except for the SCAD group, the remaining 130 cases were the control group(n=130). The differences of RDW levels between the two groups were compared, and the correlation between RDW and severity of coronary artery disease (Gensini score) and independent risk factors for SCAD were analyzed. Results The RDW level in the SCAD group was higher than that in the control group (P<0.05). There was a positive correlation between the RDW and Gensini scores in the SCAD group (r=0.217,P=0.002). Multivariate logistic regression analysis showed RDW (OR=2.950, 95% CI: 1.986~4.381, P<0.001) is an independent risk factor for SCAD. Conclusion RDW is positively correlated with the severity of coronary artery disease in SCAD. RDW is independent risk factor of SCAD and provides predictive value for the diagnosis of SCAD.

目的 探讨钙调磷酸酶结合蛋白1(calcineurin binding protein 1, Cabin1)在肾小管上皮细胞(renal tubular epithelial cells,RTECs)线粒体损伤中的作用机制。方法 采用siRNA干预体外培养RTECs,敲低Cabin1的表达,继而以电镜观察其对RTECs线粒体形态的影响。结果 在对照组和阴性对照组中Cabin1蛋白在RTECs中有高表达,采用siRNA干预RTECs后,Cabin1蛋白的表达量较对照组和阴性对照组降低50%以上(P＜0.05)。对照组与阴性对照组中,线粒体形态规则,呈圆形或椭圆形,线粒体膜完整,线粒体嵴清晰可见。敲低组中,线粒体肿胀,呈长条形或不规则形,线粒体膜、线粒体嵴结构模糊甚至消失。结论 敲低Cabin1引起RTECs的线粒体形态学异常,提示Cabin1是维持RTECs线粒体正常功能的重要分子。

Objective To investigate the role of calcineurin binding protein 1 (Cabin1) in renal tubular epithelial cells (RTECs) mitochondrial dysfunction. Methods Knocked down Cabin1 in RTECs with siRNA, Western bolt was applied to detect the level of Cabin1 protein. The morphology of mitochondria in RTECs were observed under microscopy. Results In control and negative control groups, Cabin1 protein was obviously expressed in RTECs. After knocked down by siRNA, Cabin1 protein expression was decreased (P＜0.05). In Cabin1 knocked down group, mitochondria changed from large and ellipsoid shape to the small, long and irregulars. Morover, mitochondria were swollen and cristae were remarkably dissolved. Conclusion Knocked down Cabin1 induced RTECs mitochondrial dysfunction, which indicates Cabin1 is a crucial factor regulating mitochondrial function.

目的 探讨Meis1在人子宫内膜细胞中的表达及其受雌、孕激素调控的规律。方法 通过免疫细胞化学和western blot的方法检测雌、孕激素对体外培养的在子宫内膜基质细胞(ESC)及Ishikawa细胞中Meis1的表达及调控。结果 Meis1在ESC和Ishikawa细胞均有表达,且均表达于细胞核中;在ESC中,E2、P4和 E2＋P4三组中Meis1平均蛋白表达水平均高于对照组(P<0.05)。Meis1在E2、P4和 E2＋P4组之间的表达水平亦差异有统计学意义(P<0.05),表达强度E2＋P4组>P4组>E2组;在Ishikawa细胞中,E2、P4和 E2＋P4使Meis1表达增强,表达强度P4组>E2＋P4组>E2组,但与对照组比较无差异(P>0.05),E2、P4和 E2＋P4各组间亦无差异(P>0.05)。结论 转录因子Meis1在ESC和Ishikawa细胞中受到雌、孕激素的调控,可能在子宫内膜容受性分子网络的构建中发挥着重要的作用。

Objective To investegate the expression and regulation discipline of Meis1 in human ESC and Ishikawa cells in vitro by estrogen and progesterone stimuli. Methods Immunocytochemistry and western blot were used to detect the expression and regulation discipline of Meis1 in human normal endometrial cells. Results Meis1 expressed both in endometrial stromal cells (ESC) and in ishikawa cells, and both situ in nucleus. In ESC, the expression of Meis1 was up-regulated by E₂/P₄ and E₂＋P₄, and the up-regulated effect may be superposition by E₂＋P₄, the differences between the groups were statistically difference(P<0.05). In Ishikawa cells, western blot showed that the expression of Meis1 was up-regulated by E₂/P₄ and E₂＋P₄. The differences weren't statistically significant when compared with the control group or between themselves(P>0.05). Conclusion The expression of transcription factor Meis1 is regularly regulated by estrogen and progesterone, which may be a key role during the formation of endometrial receptivity molecular network.

目的 观察重组牛碱性成纤维细胞生长因子应用于治疗浅Ⅱ度烧伤创面的临床效果。方法 选取90例小面积浅Ⅱ度烧伤患者,随机平均分为2组:应用重组牛碱性成纤维细胞生长因子治疗的患者为治疗组,使用碘伏油纱治疗的患者为对照组,观察创面愈合时间、患者疼痛程度及远期色素沉着、瘢痕增生几率情况。结果 重组牛碱性成纤维细胞生长因子治疗组效果优于对照组,治疗组创面愈合时间(8.56±2.51)d短于对照组(12.42±2.13)d(P﹤0.05),平均愈合时间较提前3~5 d;患者疼痛度减轻[VAS评分分别是:(1.66±0.05)和(3.25±0.12),P﹤0.05];部分患者一年后随访发现治疗组远期色素沉着较轻,疤痕增生几率低。结论 应用重组牛碱性成纤维细胞生长因子治疗小面积浅度烧伤能够缩短创面愈合时间,在有效促进烧伤创面愈合同时,可减轻换药时疼痛,减轻远期色素沉着,降低瘢痕增生率。

Objective To observe effect of the recombinant bovine basic fibroblast growth factor in the treatment of superficial Ⅱ degree burn wounds. Methods Ninety cases of small area of superficial Ⅱ degree burn wounds were randomly divided into 2 groups: recombinant bovine basic fibroblast growth factor group(rb-bFGF) and control group (Iodophor gauze group). The wound healing time,patient pain, long-term hyperpigmentation and scar chance of proliferation were observed. Results The effect of rb-bFGF treatment group was better than that of control group. The healing time of the treatment group was (8.56 ± 2.51)d, it was shorter than that of the control group (12.42±2.13)d(P<0.01). The average healing time was 3~5 days, it was ahead of the control group; Compared to the control group, the rb-bFGF group had less pain (VAS scores were: 1.66±0.05 and 3.25±0.12,P<0.01); Some patients were followed up a year later, we found that the treatment group long-term pigmentation was lighter, scar chance of proliferation was lower. Conclusion Application of rb-bFGF in the treatment of small area of shallow Ⅱ degree of burns may shorten the wound healing time. As effective promotion of burn wound healing, it may reduce the pain when dressing, reduce long-term pigmentation and scarring rate.

目的 本研究从细胞生物学角度检测二甲双胍对小鼠胰岛瘤MIN6的影响,并探讨此过程中包含的分子生物学机制。方法 MTT法检测不同浓度二甲双胍(1、2、5、10、20 mmol/L)对MIN6细胞活力的影响,细胞划痕实验检测二甲双胍对MIN6细胞迁移的影响,免疫印记实验检测此过程中细胞凋亡相关蛋白Bcl-2、Bax、caspase3表达的变化,及AMPK和JNK信号通路蛋白磷酸化水平的变化。结果 二甲双胍浓度大于10 mmol/L时可以抑制MIN6细胞的活力(P<0.01),降低其迁移能力(P<0.01),高浓度二甲双胍可以上调细胞内凋亡蛋白Bax(P<0.05)和p-AMPK的表达(P<0.05),降低抗凋亡蛋白Bcl-2的表达,增加caspase3剪切体(P<0.05)。同时,二甲双胍可以降低MIN6细胞内JNK信号通路的磷酸化水平(P<0.05)。结论 高浓度二甲双胍可以抑制MIN6细胞的增殖和迁移,其作用可能与降低了JNK信号的通路活化有关。

Objective This study aims to investigate the effect of metformin on proliferation and migration of MIN6 cells, and to explore the underlying mechanism. Methods The viability of MIN6 cells that were treated with various metformin (1,2,5,10 and 20 mmol/L) was detected by MTT assay. The migration of MIN6 cells was determined by wound-healing assay. Meanwhile, the proteins expression of Bcl-2, Bax, caspase3 and the phosphorylation of AMPK, JNK was detected by western bolt assay. Results The cell viability and the migration of MIN6 cells were decreased when the concentration of metformin above 10 mmol/L(P<0.01). The expression of apoptosis-related protein Bax(P<0.05) and p-AMPK(P<0.05)was up-regulated, anti-apoptosis-related protein Bcl-2 was down-regulated and cleaved caspase3 (P<0.05)was increased after high metformin treatment. At the same time, the phosphorylation of JNK was down-regulated by metformin(P<0.05). Conclusion High concertration of metformin may inhibit the proliferation and migration of MIN6 cells through suppressing the activation of JNK signaling pathway.