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 探讨前列地尔联合依帕司他对糖尿病足患者创面肉芽组织肿瘤坏死因子(tumor necrosis factor-α,TNF-α)、白介素-6 (interleukin-6,IL-6)及血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的影响。方法 将90例糖尿病足患者随机分为研究组和对照组,每组45例,对照组予常规治疗,研究组在常规治疗基础上给予前列地尔+依帕司他联合治疗。监测两组患者创面愈合率,患肢足背血流动力学及腓总神经传导速度,创面肉芽组织TNF-α、IL-6、VEGF含量及基因表达变化。结果 治疗后第2、4周研究组较对照组创面愈合率升高,差异有统计学意义(P<0.05);治疗后两组患者足背动脉血流动力学及腓总神经传导速度均有改善,而研究组疗效更明显(P<0.05);治疗后研究组患者创面肉芽组织TNF-α、IL-6含量及基因表达较对照组降低,VEGF含量及基因表达则升高(P<0.05)。结论 前列地尔联合依帕司他联合治疗可改善糖尿病足患者足背动脉血流动力学,促进受损神经功能恢复;降低糖尿病足患者创面肉芽组织TNF-α、IL-6表达,减轻免疫损伤;增加VEGF基因表达,促进血管生成,加速创伤愈合。

Objective To investigate the effects of alprostadil combined with epalrestat on the expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF) in granulation tissue in patients with diabetic foot patients.Methods Totally 90 cases with diabetic foot were randomly divided into study group (45 cases)and control group(45 cases). The control group received conventional treatment for 4 weeks and the study group additionally received combination of alprostadil and epalatone for 4 weeks. The curative rate of wound healing, limb dorsal hemodynamics, peroneal nerve conduction velocity and the levels of TNF-α, IL-6 and VEGF in the granulation tissue of the wound were monitored in the two groups.Results The wound healing rate of the study group was significantly higher than that of the control group at the 2nd and 4th week after treatment (P<0.05). After treatment, the hemodynamics and peroneal nerve conduction velocity were improved (P<0.05). The contents and expressions of TNF-α and IL-6 in the granulation tissue of the treatment group were significantly lower than that of the control group, and the content of VEGF and gene expression were significantly increased in the study group (P<0.05).Conclusion The combination therapy of alprostadil and epalrestat may improve the hemodynamics of dorsalis pedis artery in patients with diabetic foot and promote the recovery of damaged nerve function.Also it may reduce the expression of TNF-α and IL-6 in the granulation tissue of diabetic patients and reduce the immune injury. It promotes angiogenesis and accelerates wound healing by increasing VEGF gene expression.

目的 探讨p27^Kip1蛋白的表达与梅州地区客家人食管鳞癌的关系。方法 收集我院病理科2003年1月—2008年12月的61例经手术切除的中国梅州地区客家人食管鳞癌组织及32例食管良性肿瘤组织为研究对象并包埋成蜡块。采用免疫组织化学技术检测组织中p27^Kip1蛋白的表达,结合患者的临床病理资料和随访资料,进行回顾性分析,并作出评价。结果 P27^Kip1蛋白在食管鳞癌细胞核和/或细胞浆都有表达。胞核表达阳性率为27.87%,低于食管良性肿瘤组织胞核表达（50.0%）（P<0.05）;而胞浆表达阳性率为49.18%,高于食管良性肿瘤组织胞浆表达（12.5%）（P<0.05）。无淋巴结转移的食管鳞癌p27^Kip1蛋白胞核表达阳性率为42.41%,高于有淋巴结转移者的阳性率（10.7%）,差异有统计学意义（P<0.05）;而无淋巴结转移的食管鳞癌p27^Kip1蛋白胞浆表达阳性率为42.4%,比有淋巴结转移者（57.12%）的阳性率有降低的趋势（P>0.05）。Ⅱ期（ⅡA+ⅡB）和Ⅲ期p27Kip1蛋白胞核表达阳性率分别为42.41%和10.71%,差异有统计学意义（P<0.05）;p27^Kip1蛋白胞浆表达阳性率有随着TNM分期的增高而增高的趋势（P>0.05）。结论 P27^Kip1蛋白作为肿瘤抑制因子,其在食管鳞癌的胞核和胞浆表达阳性呈相反趋势,随食管鳞癌TNM分期越高,胞核阳性率越低,胞浆阳性率越高,其可作为食管肿瘤恶性程度及进展的预测指标。

目的 研究鳞癌抗原(SCC-Ag)的表达对宫颈癌手术治疗后临床转归的预测价值。方法 选取我院2014年4月—2015年9月实施宫颈癌手术的患者52例,分别于手术前后分析所有患者的鳞癌抗原表达,对患者进行1年的随访,以发生癌细胞转移、复发或死亡为研究终点,比较预后良好的患者与预后差的患者鳞癌抗原的表达的不同。结果 Ia、Ib1、Ib2、IIa、IIb期宫颈癌患者术后SCC-Ag水平较手术前均显著降低(P＜0.05);纳入本次研究的患者宫颈癌术后复发或转移发生率为15.38%,转归良好的患者为84.62%,预后良好的患者术后SCC-Ag水平(0.91±0.27)ng/mL较发生复发或转移的患者(1.37±0.57)ng/ml显著较较低(P＜0.05)。结论 宫颈癌患者术后血清SCC-Ag水平与肿瘤的应答之间具有关联性,术后SCC-Ag水平高的患者复发与转移发生率显著高于SCC-Ag水平低的患者,鳞癌抗原的表达对宫颈癌手术后患者的转归情况具有预测价值,临床应予以重视。

目的 利用大肠杆菌原核表达系统优化表达纯化肠道病毒71型(EV71)VP3结构蛋白,为后续单克隆抗体制备及检测试剂盒研发提供前期基础。方法 采用PCR方法扩增EV71病毒VP3基因,将其插入表达载体pET28a(+),构建pET28a-VP3重组质粒,转化大肠杆菌BL21(DE3)菌株,分别在25 ℃、37 ℃下经IPTG诱导表达,重组表达的蛋白产物经凝胶电泳初步分析,比较不同温度诱导表达的蛋白产物。结果 成功构建pET28a-VP3重组质粒,不同温度下诱导表达的蛋白产物在30.5 kDa左右位置均出现目的条带;37 ℃下诱导表达的蛋白超声破碎并离心后,目的蛋白基本位于沉淀中,而25 ℃诱导表达的蛋白产物有少量目的蛋白溶解于上清液中。结论 在25 ℃或37 ℃下均能利用大肠杆菌原核表达系统有效表达EV71病毒VP3蛋白;37 ℃诱导时蛋白可融性表达低,目的蛋白获取效率较高。

Objective To express VP3 capsid protein of enterovirus 71 by using Escherichia coli prokaryotic expression system. Methods VP3 gene was amplified by PCR before inserted into pET28a(+) plasmid. Then the plasmid pET28a-VP3 was transformed and expressed in the Escherichia coli BL21 strain at 25 ℃ or 37 ℃. Finally the protein was analyzed by SDS-PAGE gel electrophoresis. Results The pET28a-VP3 plasmid was successfully constructed, and the EV71 VP3 protein was expressed. Supernatant of the production after ultrasonication and centrifugation got a little VP3 protein. Conclusion The EV71 VP3 protein was expressed. Expression at 25 ℃ may lead to the dissolution of the recombinant protein.

目的 观察芳香烃受体(AhR)及Th17相关细胞因子在类风湿关节炎中的表达水平及其对疾病的预测价值。方法 选择2014年1月—2015年12月于我院就诊的RA患者60例作为观察组,选取同期于我院进行健康体检的正常人60例作为对照组,采用酶联免疫吸附法检测血清中 IL-17、IL-23及AhR的表达水平,并分析其与疾病活动度的关系。结果 RA患者血清IL-17、IL-23及AhR水平高于对照组(P＜0.05)。而根据病情严重程度,RA 非早期组IL-17、IL-23及AhR水平较早期组增高(P＜0.05)。血清 IL-17 水平与除CRP以外的病情活动度指标均呈正相关关系(P＜0.05)。IL-23 水平与SJC、TJC、HAQ 、DAS28 评分呈正相关关系(P＜0.05),但其他指标无明显相关性(P>0.05)。RA患者PBMC中AhR的表达水平与各项临床指标无相关性(P>0.05)。IL-17和IL-23水平与Sharp评分呈正相关关系(r=0.895,P＜0.01;r=0.708,P＜0.01)。AhR的表达与血清IL-17和IL-23水平呈正相关(r=0.415,P＜0.01)。经荧光定量PCR检测结果显示,RA患者AhR及其下游因子CYP1A1、IL-17、FOXP3 mRNA的表达水平高于对照组(P＜0.05)。且RA 非早期组AhR及其下游因子CYP1A1、IL-17、FOXP3 mRNA的表达水平较早期组增高(P＜0.05)。经相关关系检测,RA患者AhR及其下游因子CYP1A1、IL-17、FOXP3 mRNA的表达水平与Sharp评分呈正相关关系(r=0.715,P＜0.01;r=0.734,P＜0.01;r=0.812,P＜0.01;r=0.755,P＜0.01)。结论 Th17相关细胞因子IL-17和IL-23在RA病理生理过程中发挥重要作用,其表达增高,提示关节炎症处于活跃状态,骨质破坏较重,可作为评估RA病情的重要指标。RA患者体内AhR蛋白及其相关下游信号通路均呈高表达状态,AhR通路在RA患者的发病过程中可能发挥关键作用。

Objective To observe the expression level of aroma receptor (AhR) and Th17 related cytokines in rheumatoid arthritis and its predictive value to disease. Methods Sixty patients with RA who were treated in our hospital from January 2014 to December 2015 were selected as the observation group. Sixty healthy subjects were randomly selected as the control group. IL-17, IL-23 and AhR levels in serum were detected by enzyme-linked immunosorbent and the relationship between IL-17, IL-23 and disease activity were analyzed. Results IL-17, IL-23 and AhR levels in serum of RA patients were significantly higher than those in controls (P<0.05). However, the levels of IL-17, IL-23 and AhR levels were significantly higher in the non- early RA group than in the early group (P<0.05), depending on the severity of the disease. There was a positive correlation between serum IL-17 level and disease activity index except CRP (P<0.05). IL-23 level was positively correlated with SJC, TJC, HAQ and DAS28 scores (P<0.05), but no significant correlation was found between other indexes (P>0.05). The expression level of AhR in PBMC of RA patients was not correlated with clinical indexes (P>0.05). IL-17 and IL-23 levels were positively correlated with Sharp score (r=0.895, P<0.01; r=0.708, P<0.01). The expression of AhR was positively correlated with serum IL-17 and IL-23 levels (r=0.415, P<0.01). The expression of AhR and its downstream factors CYP1A1, IL-17 and FOXP3 mRNA in RA patients were significantly higher than those in the control group (P<0.05) by fluorescence quantitative PCR. The expression of AhR and its downstream factors CYP1A1, IL-17 and FOXP3 mRNA in RA group were significantly higher than those in early group (P<0.05). The expression level of AhR and its downstream factors CYP1A1, IL-17 and FOXP3 mRNA in RA patients were positively correlated with Sharp scores (r=0.715, P<0.01; r=0.734, P<0.01; r=0.812, P<0.01; r=0.755, P<0.01). Conclusion The Th17-related cytokines IL-17 and IL-23 play an important role in the pathophysiology of RA, and the expression of Th17-associated cytokines is increased, suggesting that arthritis is active and bone destruction is serious. The AhR protein and its associated downstream signaling pathways are highly expressed in RA patients, and the AhR pathway may play a key role in the pathogenesis of RA patients.

目的 观察乳腺癌组织中沉默信息调节因子(SIRT1)的表达并探讨其临床意义。方法 选取70例乳腺癌组织和20例乳腺纤维腺瘤组织,应用免疫组化法观察SIRI1在两组中的表达,并分析其与临床病理参数、预后之间的关系。结果 SIRT1在乳腺癌组织中和乳腺纤维腺瘤组织中的阳性表达分别为:62.9%(44/70)、35%(7/20),P＜0.05;SIRT1的表达与乳腺癌的病理分期及淋巴结转移有关,P＜0.05;SIRT1表达阳性和阴性患者2年无疾病进展生存率分别为40.6%和73.7%,P＜0.05。结论 乳腺癌组织中SIRT1的阳性表达率较高,并与乳腺癌的发生、发展、侵袭、转移、预后有关。

Objective To investigate the sirtuin (SIRT1) expression in breast cancer and its relationship with clinicopathological parameters in breast tissue. Methods We used immunohistochemical staining (Envision two-step) to detect the expression of 70 cases of breast cancer and 20 cases of breast fibroadenoma of SIRT1. Results The positive expression of sirt1 in breast cancer and breast fibroadenoma tissues were 62.9 % (44/70) and 35 % (7/20),P<0.05. The expression of sirt1 was correlated with the pathological stage and lymph node metastasis of breast cancer, P<0.05. The 2-year survival rates of sirt1 positive and negative patients were 40.6 % and 73.7 %, respectively, P<0.05. Conclusion The positive expression rate of sirt1 in breast cancer is higher, and is related to the occurrence, development, invasion, metastasis and prognosis of breast cancer.

目的 探讨川芎嗪对链脲佐菌素(STZ)诱导2型糖尿病大鼠肾病的治疗作用及机制。方法 SD大鼠50只,随机分为正常组和模型组。除正常组外,其余大鼠均给予高脂-高糖饲料喂养4周,再给予链脲佐菌素(40 mg/kg,ip),72 h后测定空腹血糖,将血糖值高于16.67 mmol/L的大鼠随机分成4个组即模型组,二甲双胍阳性组(250 mg/kg),川芎嗪低、高剂量组(80、160 mg/kg),连续给予相应试药8周。其中正常组和模型组的大鼠均给予同等量蒸馏水灌胃。实验结束时,测定大鼠血糖、尿蛋白、血尿素氮和血肌酐含量;免疫组化法测定大鼠肾组织TLR4和caspase3蛋白表达。光镜下观察肾脏病理学变化。结果 与模型组比较,二甲双胍组和川芎嗪高剂量组给药8周后,大鼠动态空腹血糖均能明显降低(P<0.05),大鼠动态尿蛋白显著性降低(P<0.01,P<0.05); 二甲双胍和高剂量组TLR4和caspase3蛋白表达明显低于模型组(P<0.05);肾脏组织病理性损伤明显减轻。结论 川芎嗪对STZ诱导2型糖尿病大鼠肾病具有保护作用,其机制可能与下调TLR4表达作用有关。

Objective To investigate the therapeutic effects and mechanisms of tetramethylpyrazine (TMP) on streptozocin(STZ)-induced-nephropathy in type 2 diabetic rats. Methods 50 SD rats were randomly divided into normal group(n=10) and model group(n=40). The model rats were fed on high fat and sugar diets for 4 weeks, then given STZ(40 mg/kg,ip). After 72 hours, the fasting blood glucose (FBG) was measured. Rats with high FBG above 16.67mmol/L were randomly divided into four groups: model, metformin(Met, 250 mg/kg)and TMP (80 mg/kg, 160 mg/kg) groups for treating 8 weeks, and both the control and model groups were given equals distilled water by intragastric administration. At the end of the experiment, blood glucose, urine protein, blood urea nitrogen and creatinine were measured. The expression of TLR4 and caspase3 protein in kidney tissue of rats was determined by immunohistochemistry. Pathological changes of kidney were observed under light microscope. Results Compared with the model group, metformin and high dose of TMP administered after 8 weeks, rats can significantly reduce the dynamic fasting blood glucose(P<0.05). Urinary protein excretion of total dynamic decreased significantly (P<0.01, P<0.05); the protein expression of TLR4 and caspase3 in the metformin group and high dose group was significantly lower than that in the model group (P<0.05); kidney tissue pathological damage was significantly reduced. Conclusion TMP has a protective effect on STZ induced nephropathy in type 2 diabetic rats, and its mechanism may be related to the down-regulation of TLR4 expression.

目的 分析PBK在前列腺癌中的表达及临床意义。方法 利用前列腺癌的组织芯片,包含98例前列腺癌及81例对照癌旁组织作为研究对象,免疫组化方法检测PBK的表达情况,并运用统计学方法分析免疫组化芯片及Taylor数据库中PBK表达与前列腺癌临床病理特征之间的关系。结果 PBK在前列腺癌中表达明显升高(P=0.001);且在Gleason高评分组的表达比低评分组表达升高(P=0.001)。Taylor数据库得到相似结果,且运用Kaplan-Meier分析发现PBK与无生化复发生存率显著相关(P=0.007),最后采用Cox回归模型进行多因素综合分析发现在影响前列腺癌预后的队列中,PBK高表达(P=0.041)与Gleason评分、病理分期都是前列腺癌生化复发的独立预测指标。结论 PBK的表达与前列腺癌密切相关,可作为临床诊断及治疗的分子标志物。

Objective To investigate the expression and clinical significance of PBK in prostate cancer. Methods Using tissue microarrays of prostate cancer, which including 98 cases of prostate cancer and 81 cases of normal tissue adjacent to cancer as the research object, the expression of PBK was detected by immunohistochemistry, and statistical analysis was used to analyze the relationship between the expression of PBK and the clinicopathological features of prostate cancer in the microarray and Taylor database. Results The expression of PBK in prostate cancer was significantly higher (P=0.001), and the expression increased in the group of high Gleason score (P=0.001). The Taylor database obtained similar results, and Kaplan-Meier analysis showed that PBK was significantly correlated with the biochemical recurrence free survival (P=0.007). Finally, Cox regression model was used to analyze the prognostic factors of prostate cancer. Result shows that, the high expression of PBK (P=0.041), Gleason score and pathological stage were independent predictors of biochemical recurrence of prostate cancer. Conclusion The expression of PBK is closely related to prostate cancer, and can be used as a molecular marker for clinical diagnosis and treatment.

目的 探讨ERCC1、RRM1、TS蛋白表达对晚期非小细胞肺癌(NSCLC)个体化治疗的指导意义。方法 收集经病理确诊的晚期NSCLC患者87例,其中67例愿意接受药敏免疫组化检测的患者作为研究组,采用SP法检测肿瘤组织ERCC1、RRM1、TS蛋白表达,并根据蛋白表达情况选择化疗方案;另外20例患者不进行药敏免疫组化检测,以常规吉西他滨联合顺铂方案化疗,以此作为对照组。比较两组患者化疗的有效率,疾病控制率(DCR),并以无进展生存期(PFS)为指标比较患者预后。结果 研究组67例患者中,PR 33例(49.25%),SD 13例(19.4%),PD 21例(31.35%);对照组20例患者中,PR 4例(20%),SD 4例(20%),PD 12例(60%),两组疗效之间有差异( χ²=6.437,P=0.04),研究组DCR为68.6%,高于对照组DCR 40%,差异有统计学意义(χ²=5.372,P=0.034)。研究组患者的中位PFS高于对照组,研究组的PFS为5月,对照组为3月,差异有统计学意义(P＜0.05)。结论 对晚期NSCLC患者进行ERCC1、RRM1、TS药敏蛋白免疫组化检测,指导个体化治疗方案,能提高患者化疗的疾病控制率及延长患者的疾病进展时间。