目的 探讨不稳定型心绞痛伴左心收缩功能不全患者血清脑钠肽前体(Pro-BNP)、同型半胱氨酸(Hcy) 和血沉(ESR)的表达与临床意义。方法 选取2015年1月—2016年10月于广州市第一人民医院心血管内科就诊的130例不稳定型心绞痛(A组) 、130例不稳定型心绞痛伴左心收缩功能不全(B组)患者作为研究对象,同时选取同期130例健康体检者(C组)作为对照。分别检测3组受试者的血清Pro-BNP、Hcy、ESR水平,并用方差分析对3组血清水平进行比较;通过Pearson相关分析比较B组患者的血清Pro-BNP、Hcy和ESR水平与左心室射血分数(LVEF)的相关性。结果 不稳定型心绞痛患者和不稳定型心绞痛伴左心收缩功能不全患者的Pro-BNP、Hcy、ESR水平均明显高于对照组,不稳定型心绞痛伴左心收缩功能不全患者的Pro-BNP、Hcy、ESR水平明显高于不稳定型心绞痛患者血清水平。随着心功能分级的升高,患者的血清Pro-BNP、Hcy和ESR水平呈明显上升趋势。血清Pro-BNP、Hcy、ESR水平与 LVEF 呈负相关,血清 Hcy、ESR水平与Pro-BNP水平呈正相关。结论 不稳定型心绞痛伴左心收缩功能不全患者血清Pro-BNP、Hcy、ESR水平与心功能分级明显相关,可作为不稳定型心绞痛伴左心收缩功能不全病情评估及预后判断的重要指标。

Objective To investigate the expressions and clinical significances of atrial brain natriuretic peptide precursor (PRO-BNP), homocysteine (Hcy) and erythrocyte sedimentation rate (ESR) in patients with unstable angina pectoris associated by Left ventricular systolic insufficiency. Methods A total of 130 patients with simple unstable angina pectoris and another 130 patients with unstable angina pectoris associated by left ventricular systolic insufficiency treated during January 2015 and October 2016 were selected as group A and group B respectively,and at the same period,130 healthy persons taking medical examination were selected as the control froup (group C). Serum levels of PRO-BNP,Hcy and ESR were detected in three groups,and the correlation between serums PRO-BNP,Hcy and ESR and left ventricular ejection fraction (LVEF) in group B were analyzed. Results Serum levels of in group A and B were significantly higher than those in group C,and the levels in group B were significantly higher than those in group A. In group B,serum levels of PRO-BNP,Hcy and ESR were significantly increased with rising cardiac function classification. Serum levels of PRO-BNP,Hcy and ESR in group B were negatively correlated with LVEF,but serum levels of Hcy and ESR were positively correlated with PRO-BNP level. Conclusion Serum levels of PRO-BNP,Hcy and ESR in group A and B are significantly correlated with cardiac function in patients with unstable angina pectoris associated by left ventricular systolic insufficiency,so the levels may be used as important indexes for evaluating the severity and prognosis of with unstable angina pectoris associated by left ventricular systolic insufficiency.

目的 观察表皮生长因子(EGF)在新生儿坏死性小肠结肠炎(NEC)患儿肠组织中的动态表达情况,探讨EGF在NEC病程中起到的保护作用。方法 选取15例NEC患儿行一期回肠造瘘手术治疗的回肠组织为实验组(NEC组),将以上15例NEC患儿行二期回肠封瘘手术治疗的回肠组织为对照组(封瘘组),采用免疫组织化学技术检测,通过光密度测算软件(IPP)分析回肠组织中的EGF表达。结果 EGF主要表达于肠壁黏膜层,少量表达于黏膜下层、肌层。EGF在NEC组各层表达平均光密度值为:黏膜层(0.241±0.075),黏膜下层(0.213±0.061),肌层(0.1397±0.026),差异有统计学意义(P＜0.05);在封瘘组各层表达情况为:黏膜层(0.211±0.028),黏膜下层(0.119±0.022),肌层(0.097±0.007),差异有统计学意义(P＜0.05)。EGF在NEC组总体表达平均光密度值为(0.198±0.071),明显高于封瘘组(0.146±0.058),差异有统计学意义(P＜0.05)。结论 表皮生长因子(EGF)在新生儿坏死性小肠结肠炎(NEC)肠组织中的表达较封瘘组显著上调,推测EGF可能与NEC炎症相关,可能在NEC炎症过程中起到了一定的保护作用。

Objective We realized that EGF could play an important protective role against NEC. However, the practical condition of the distribution and expression of EGF in intestine of infants with NEC was indefinite. In order to figure out this problem,we carried out this experimentation. Methods The sample were divided into two group.The experimental group(necgroup) were composed of fifteen individual intestinal tissues after the ileostomy were performed on those infants suffered from NEC. The control group(sealing fistula group) were composed of fifteen individual intestinal tissues after the ileal closure fistula were performed on the same infants who were accepted the one-stage ileostomy in the period of NEC and were later accepted the two-stage operation on the condition that their bodies almost recovered from NEC after two to three months gone.Then, we utilized immunohistochemistry to test the distribution and quantities of EGF on those samples of the two group infants. Results The characteristic of EGF expression in intestine of the both group included strong positive expression in mucous layer and less expression in strata submucosum and muscular coat. The average optical density in nec group was mucous layer (0.241±0.075),strata submucosum(0.213±0.026),muscular coat (0.1397±0.022);In the control groupmucous layer (0.211±0.028),strata submucosum (0.119±0.022),muscular coat (0.097±0.007). The expression of EGF in intestinal tissues increased in the period of NEC0.198±0.071 by comparing with the control group (0.146±0.058). Conclusion There may be a correlation between the strong positive expression of EGF in intestinal tissues in the period of NEC and inflammation.By combining the result of this experiment and the research about EGF. We assumed that EGF is one factor of the protective mechanism by which injured intestinal mucous could be recovered and resist inflammation.

目的 观察比较术中及术前化疗干预对进展期胃恶性肿瘤手术患者p53、ki-67表达及预后的影响,为临床化疗时间的选择提供理论依据。方法 自2014年8月—2015年5月,我院共收入胃恶性肿瘤患者40例,将40例患者随机分为两组,每组各20例,保证两组患者在性别、年龄、胃癌分期等方面可比,无统计学差异(P＞0.05),标记为Ⅰ组和Ⅱ组。Ⅰ组20例患者于术前进行化疗干预,Ⅱ组在术中给予化疗干预。观察比较两组患者p53、ki-67表达状况及预后。结果 Ⅰ组及Ⅱ组治疗后p53及ki-67均比治疗前升高,差异有统计学意义(P<0.05)。但是治疗后,Ⅰ组和Ⅱ组的p53表达状况组间差异不明显,无统计学意义(P>0.05)。治疗前后,AI差异有统计学意义(P<0.05)。Ⅰ组效果明显好于Ⅱ组,两者差异有统计学意义(P <0.05)。术后六个月、一年随访时发现两组复发率、死亡率差别不大,无统计学意义(P>0.05),术后两年随访发现Ⅱ组复发率、死亡率明显低于Ⅰ组,差异有统计学意义(P<0.05)。结论 术中化疗的疗效优于术前化疗,患者预后较术前化疗好。

Objective To observe the effect of intraoperative and preoperative chemotherapy on the expression of p53, Ki-67 and prognosis in patients with advanced gastric cancer. Methods 40 cases of advanced gastric cancer in our hospital from Aug 2014 to May 2015 were enrolled in the study, and were divide into 2 groups randomly. In group I, 20 patients received chemotherapy intervention befoerer operation, and the other group received chemotherapy intervention during operation. The expressions and prognosis of p53 and Ki-67 were observed and compared between the two groups. Results Group Ⅰ and group Ⅱ after treatment, p53 and Ki-67 were higher than that before treatment, with statistical significance(P<0.05). However, there was no significant difference in the expression of p53 between group Ⅰ and group Ⅱ after treatment, and there was no significant difference(P>0.05). Before and after treatment, the difference of AI was significant, with statistical significance (P<0.05). The effect of group Ⅰ was obviously better than that of group Ⅱ, the difference was statistically significant(P<0.05). Six monthse after the operation and one year follow-up found two groups of recurrence rate and mortality rate had no significant difference(P>0.05). After two years follow-up found the group Ⅱ recurrence rate, mortality was lower than in group Ⅰ (P<0.05). Conclusion The effect of intraoperative chemotherapy is better than that of preoperative chemotherapy, and the prognosis is better than that of preoperative chemotherapy.

目的 了解DFU患者的TXA2表达水平的变化及其影响因素,并讨论其与糖尿病血管病变的严重程度的相关性。方法 选择我院内分泌科2009年6月—2012年12月收治的2型糖尿病足部溃疡并坏疽形成的患者共30例、DFU非坏疽组38例、无DFU的2型糖尿病组患者40例,收集其一般资料及检查结果。分析TXA2的表达水平与其他资料的关系。并采用相关分析TXA2的稳定代谢产物TXB2水平升高的危险因素。结果 年龄、糖尿病病程、吸烟、BMI、血浆白蛋白与TXB2水平相关。结论 2型糖尿病足部溃疡患者的血浆血栓素A2水平显著增高,并且表达水平与病情严重程度相关,且与糖尿病病程、血脂、血糖 、血胰岛素、胰岛素抵抗指数水平、血压控制水平等呈明显相关关系。提示其可作为 2 型糖尿病患者血管内皮功能及动脉硬化的早期反映。

目的 探讨小脑延髓池注射纳洛酮对心肺复苏大鼠脑神经保护的作用机制。方法 将30只雄性SD大鼠随机分为假手术组、常规复苏组和纳洛酮复苏组。采用窒息法建立大鼠心脏骤停模型,复苏的同时给予药物治疗。恢复自主循环(ROSC)后24 h取脑组织,荧光定量PCR法检测脑组织c-Fos mRNA表达水平,免疫组化法检测脑组织c-Fos蛋白的表达。结果 与常规复苏组比较,纳洛酮可显著降低大鼠脑组织c-Fos mRNA及蛋白表达量(P<0.01)。结论 小脑延髓池注射纳洛酮可及时有效的作用于c-Fos基因,发挥脑神经保护作用。

Objective To investigate the neuroprotective mechanism of naloxone injected into cisterna magna on cerebral ischemia-reperfusion. Methods Thirty adult male SD rats were randomly divided into sham group, conventional cardiopulmonary resuscitation (CPR) group and naloxone CPR group. Asphyxiation was used to set up rat cardiac arrest model, and corresponding drugs were given when the resuscitation was carried out. The Brain tissues were taken at 24 h after restoration of spontaneous circulation(ROSC). Fluorescence quantitative polymerase chain reaction (PCR) and immunohistochemical was used to detect the expression of c-Fos proteins was used to detect the expression of c-Fos mRNA level. Results Compared with the conventional CPR group, Naloxone could significantly decrease the expression of c-Fos protein and c-Fos mRNA in rat brain. Conclusion Naloxone injected into cisterna magna can promptly and effectively act on c-Fos gene, playing a neuroprotective role.

目的 研究细胞周期调控因子E2F-1和p16蛋白与宫颈上皮内瘤变及宫颈鳞癌的表达及其临床意义。方法 通过免疫组化SP法检测宫颈上皮内瘤变Ⅰ-Ⅱ级(CINⅠ-Ⅱ级)、宫颈上皮内瘤变Ⅲ级(CINⅢ级)、宫颈鳞癌各40例中E2F-1和p16蛋白的表达情况,并用20例慢性宫颈炎组织作对照。结果 E2F-1蛋白在CINⅠ-Ⅱ级、CINⅢ级和宫颈鳞癌组中阳性率分别为17.5%、67.5%和80.0%, 3组间有差异(χ²=37.278,P<0.001)。p16蛋白表达阳性率分别为27.5%、70.0%和82.5%,3组间差异有统计学意义(χ²=27.708, P<0.001)。CINⅠ-Ⅱ级与CINⅢ级组及宫颈鳞癌组比较均有差异,但CINⅢ级与宫颈鳞癌组间无差异。p16蛋白的表达与E2F-1蛋白表达有正相关关系;在鳞癌组中E2F-1与p16的异常表达与组织学分级、临床分期有关(P<0.05)。结论 子宫颈鳞癌的形成与E2F-1 、p16蛋白过表达是呈正相关关系,E2F-1 、p16 蛋白可能作为子宫颈鳞癌及CIN的标志物,对子宫颈癌筛查和预防有重要意义。

Objective To explore the clinicopathologic significance of the protein expression of cell cycle regulators E2F-1 and P16 in cervical intraepithelial neoplasia(CIN) and cervical squamous cell carcinoma(SCC). Methods Immunohistochemical technique S-P was used to determinate the expression of E2F-1 and P16 protein in 40 cases with CINⅠ-Ⅱ, 40 cases with CINⅢ and 40 cases with cervical squamous cell carcinoma. Results The positive rate of E2F-1 protein expression in CINⅠ-Ⅱ, CINⅢ and cervical squamous cell carcinoma was 17.5%,67.5% and 80.0%, respectively. There were significant differences among the three groups(χ²=37.278,P<0.001). The positive rate of P16 protein expression in CINⅠ-Ⅱ, CINⅢ and cervical squamous cell carcinoma was 27.5%,70.0% and 82.5%, respectively. There were significant differences among the three groups (χ²=27.708, P<0.001). The group with CINⅢ and cervical squamous cell carcinom compared with the group withCINⅠ-Ⅱ did have differences. But there were no significant differences in the group with CINⅢ and the group with cervical squamous cell carcinoma. The relative expressions between P16 and E2F-1 showed positive correlation. In the cervical squamous cell carcinoma group, the abnormal expression of E2F-1 and p16 were correlated with the histological grade and clinical stage. Conclusion It showed positive correlation between very high expression of p16 protein and E2F-1 infection in CIN and SCC. p16 protein and E2F-1 can be an indicator in SCC and CIN,which has very significance in preventing and screening of cervical cancer.

目的 观察miR-9在胃癌组织中的表达水平及其与临床病理的相关性。方法 采用实时荧光定量PCR方法分别检测28例胃癌及正常胃组织中miR-9的表达水平,并分析其表达情况与临床病理资料的关系。结果 胃癌组织miR-9的表达水平[0.0078(0.0031~0.0142)]显著低于相应正常胃组织[0.0177(0.0084~0.0311), P<0.05],尤其是伴淋巴结转移者[0.0021(0.0006～0.0685)]miR-9表达水平明显降低。结论 胃癌组织中miR-9低表达,且与胃癌淋巴结转移有关。

Objective To investigate the correlation between the level of miR-9 expression and clinicopathology in gastric carcinoma. Methods The expression of miR-9 in 28 cases of gastric carcinoma tissues and normal gastric tissues were detected by quantitative real-time PCR, and the relationship between the expression and clinicopathology was analyzed. Results The expression of miR-9 expression in gastric carcinoma tissues [0.0078(0.0031~0.0142)] was significantly lower compared with the corresponding normal gastric tissues [0.0177(0.0084~0.0311), P<0.05], especially in lymph node-invasive carcinoma [0.0021(0.0006～0.0685)]. Conclusion miR-9 was down regulated in gastric cancer tissues, which was significantly associated with lymph node metastasis.

目的 探讨2型糖尿病肾病(DN)患者肾组织中STOML2的表达及作用。方法 免疫组化检测临床2型糖尿病肾病患者肾组织STOML2的表达及定位,采用慢病毒转染方法建立稳定过表达STOML2的HK-2细胞系,并应用Western blot检测肾小管上皮细胞钙粘蛋白(E-cadherin)、Fibronectin和STOML2的表达。结果 STOML2在DN患者肾组织的肾小管上皮细胞胞浆中表达明显升高。在高糖刺激HK-2细胞建立的EMT模型中,STOML2呈时间依赖表达上调。STOML2稳定高表达时,E-cadherin表达下调,而Fibronectin明显上调,即能促进肾小管上皮细胞发生EMT。结论 STOML2可能通过促进肾小管上皮细胞向间充质细胞分化,进而参与糖尿病肾病肾脏纤维化的发生发展。

Objective To investigate the expression and role of STOML2 in renal tissue of patients with type 2 diabetic nephropathy. Methods To detect he expression and localization of STOML2 in clinical renal tissue in patients with type 2 diabetic nephropathy by immunohistochemistry, and use lentiviral transfection method to establish a stable cell line of over-expressing STOML2, lastly apply western blot to detect the expression of E-cadherin, Fibronectin and STOML2 in renal tubular epithelial cells. Results STOML2 was significantly increased in renal tubular epithelial cytoplasm of patients with DN. In the EMT model of HK-2 cells stimulated by high glucose, STOML2 was increased in a time dependent. Overexpression of STOML2 led to E-cadherin down-regulated, while Fibronectin up-regulated, which promoted the occurrence of EMT in renal tubular epithelial cells. Conclusion STOML2 may be involved in the development and progression of renal fibrosis in diabetic nephropathy by mediating epithelial- mesenchymal transition of renal tubular epithelial cells.

目的 探讨初诊的弥漫大B细胞淋巴瘤(DLBCL)患者外周血Th17细胞的表达与国际预后指标(IPI)之间关系。方法 初诊DLBCL组(n=45)按照国际预后指数(IPI)积分分为4组,采用ELISA和流式细胞术检测各个DLBCL组与正常对照组(n=43)的外周血中IL-17的浓度以及Th17阳性细胞比例,比较各组数值间的差异,并分析IPI的5个指标与IL-17的浓度以及Th17阳性细胞比例的相关性。结果 DLBCL中高危组与高危组的IL-17的浓度以及Th17阳性细胞比例较正常对照组及其他IPI组降低,有显著性差异;DLBCL四组的IL-17的浓度以及Th17阳性细胞比例均低于正常对照组;且可见随着IPI分组的增高,IL-17的浓度以及Th17阳性细胞比例呈降低的趋势;IPI指标中年龄、临床分期、全身状态与Th17细胞的表达有相关性。结论 初诊DLBCL患者外周血Th17细胞的表达与国际预后指标有关系;随着IPI积分的增加,DLBCL患者Th17细胞表达下降;临床上对于年龄60岁以上、临床分期Ⅲ期以上、长期卧床及需别人照顾的患者更要注意监测其外周血Th17细胞的表达情况。

Objective To explore the relationship between the international prognosis indexes(IPI) and the Th17 cells expression in DLBCL patients. Methods DLBCL patients (n=45) were divided into 4 groups according to IPI score, peripheral blood were taken from each person in DLBCL groups and normal group. We used ELISA to test IL-17 and flow cytometry (FCM) to examine the Th17 positive cells. We compared the value of each group, and analyzed the relativity of IPI and Th17 cells' expression. Results Th17 cells' expression level in middle-high risk group and high risk group were higher than that in normal group and other IPI groups; Th17 cells' expression level in DLBCL groups were all lower than that in normal group; Th17 cells decreased while IPI score increased; Age, clinical stage and general body state have the relativity with DLBCL patients' Th17 cells expression level. Conclusion Th17 cells in DLBCL patients has the relativity with prognosis index. In our clinical diagnosis and treatment, we need to pay more attention to those patients who are over 60 years old, or whose clinical stage is above Ⅲ phase, or who need to stay on bed for a long time and need other peoples' help .

目的 构建重组pEGFP-C3-HCVc真核表达载体,并建立稳定表达HCVc基因的肝内胆管癌细胞株RBE-core。方法 采用PCR钓取目的基因HCVc,并克隆入pEGFP-C3的多克隆位点,构建pEGFP-C3-HCVc重组质粒。经过双酶切及测序验证后,采用脂质体将pEGFP-C3-HCVc质粒转染到RBE细胞中,经2周G418 (200 μg/mL) 筛选后进行单克隆挑选及扩大培养,建立稳定表达HCVc的胆管癌细胞株RBE-core。采用RT-PCR和Western blot验证HCVc在RBE-core中的表达情况。结果 PCR成功钓取HCVc基因,大小约573 bp,并插入pEGFP-C3载体HindⅢ和BamHⅠ多克隆位点;双酶切及测序证实目的基因HCVc正确连接到pEGFP-C3的多克隆位点。RT-PCR和Western blot分别在573 bp处和34 KD左右检测到相应的阳性条带。结论 成功构建重组质粒pEGFP-C3-HCVc,并在胆管癌细胞RBE中获得稳定表达。

Objective To construct a recombinant plasmid of pEGFP-C3-HCVc containing hepatitis C virus core protein, and establish the HCVc-expressing cell line RBE-core. Methods The HCVc gene was amplified by PCR and cloned into HindⅢ and BamHⅠsite of pEGFP-C3 plasmid. The recombinant plasmid of pEGFP-C3-HCVc was confirmed by sequencing. RBE cells were transfected with the recombinant plasmid by using Lipofectamine 2000, and then performed G418 (200 μg/mL) selection after 2 weeks. The expressing of HCVc gene in RBE cells was confirmed by RT-RCR and western blot. Results The recombinant plasmid of pEGFP-C3-HCVc was successfully constructed. RT-PCR and western blot detected a 573bp and 34KD bland, indicating the stably expressing of HCVc in RBE cells. Conclusion The recombinant plasmid of pEGFP-C3-HCVc is stabled expressing in RBE cells,which provides support for the further study.