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.

目的 研究前列腺癌细胞中miR-221的表达情况及其对癌细胞增殖的影响。方法 运用实时荧光定量PCR(qRT-PCR)检测miR-221在前列腺正常细胞株与前列腺癌细胞株中表达的差异情况,利用细胞转染构建miR-221过表达LNCaP和DU145细胞株,再通过CCK8细胞增殖实验检测细胞增殖情况的变化。结果 qRT-PCR检测细胞株发现miR-221在PC3、LNCaP和DU145三种前列腺癌细胞株中表达量均比前列腺正常细胞株PrEC低 (F=254.197,P<0.001),其中两两比较差异也均有统计学意义。细胞转染技术构建的miR-221过表达LNCaP和DU145细胞株,经qRT-PCR结果显示,miR-221在LNCaP和DU145细胞株中的表达水平明显升高(LNCaP,倍数变化=2.24,t=3.46,P<0.01;Du145,倍数变化=2.24,t=4.29,P<0.01)。细胞增殖实验结果显示,过表达了miR-221的LNCaP(P<0.001)和DU145(P<0.001)细胞生长速度慢于对照组。结论 实验证明miR-221表达过度能减慢前列腺癌细胞的增殖,miR-221有可能成为前列腺肿瘤治疗的生物学标志物。

Objective To investigate miR-221 expression in prostate cancer cells and its influence on prostate cancer cell proliferation. Methods miR-221 expressions in prostate normal cell lines and cancer cell lines were measured by qRT-PCR. Overexpression of the miR-221 in LNCaP and DU145 cell lines used by cell transfection. Effects of the depletion on cell proliferation were assessed in vitro with CCK8. Results qRT-PCR showed miR-221 was lower expressed in PC3, LNCaP and DU145 than in PrEC(F=254.197, P<0.001), in which pairwise comparison also had significant differences. qRT-PCR showed miR-221 expression rose significantly in LNCaP and DU145 cell lines whose miR-221 was overexpression with cell transfection(LNCaP, Fold Change=2.24,t=3.46,P<0.001;Du145, Fold Change=2.24,t=4.29,P<0.001). Cell proliferation assay showed that growth of LNCaP(P<0.001) and DU145(P<0.001) cells whose miR-221 was overexpression was slower than the control group. Conclusion This study demonstrates miR-221 overexpression can inhibited the proliferation of prostate cancer cells for the first time, it also suggests that miR-221 has the potential to serve as a biomarker for PCa therapy.

目的 探讨Bcl-2、COX-2在宫颈癌新辅助化疗前后表达的意义, 以及新辅助化疗(NACT)对宫颈癌的近期临床疗效。方法 对32例宫颈癌患者,采集NACT治疗前后的宫颈癌组织标本,采用免疫组织化学SP法检测组织中的Bcl-2及COX-2表达。结果 ①经NACT后,治疗总有效率(CR+PR)为75%,无效率(PD+SD)为25%。②宫颈癌组织中Bcl-2、COX-2的表达均出现明显下降,差异均有统计学意义(P<0.05);临床有效组中Bcl-2、COX-2的表达在NACT后出现显著下降(P<0.05),无效组中Bcl-2、COX-2的表达在NACT前后无明显统计学意义(P>0.05)。结论 Bcl-2、COX-2的表达情况对评价宫颈癌患者新辅助化疗效具有肯定的临床意义,宫颈癌行NACT后近期疗效良好。

Objective To investigate the expression of Bcl-2 and COX-2 in cervical cancer before and after neoadjuvant chemotherapy; To evaluate the efficacy of neoadjuvant chemotherapy(NACT) for cervical cancer in the recent clinical effects. Methods To select 32 cases of patients with cervical cancer, collect the cervical cancer tissues before and after NACT, immunohistochemical SP method was used to detect the expression of Bcl-2and COX-2 in the tissues. Results After neoadjuvant chemotherapy, total effective rate (CR+PR) was 75%(24/32), inefficient rate(PD+SD) was 25%(8/32). The expression of Bcl-2 and COX-2 of cervical cancer patients who had neoadjuvant chemotherapy (NACT), before and after, had great differences. The difference had statistical significance (P<0.05); The expression of Bcl-2 and COX-2 were significantly lower after neoadjuvant chemotherapy in clinical effective group(P<0.05), there is no statistical significance in clinical non-effective group(P>0.05). Conclusion The expression of Bcl-2 and COX-2 of cervical cancer patients has certain clinical significance in evaluating the effect of neoadjuvant chemotherapy in cervical cancer patients. Recent curative effect after NACT in the cervical cancer patients is good.

目的 探讨胃癌组织中刺猬信号通路(Hedgehog signaling pathway, Hh)中的音猬因子(Sonic hedgehog, Shh)和胶质瘤相关癌基因同源物-1(Glioma-associated oncogene homolog -1, Gli-1)与金属基质蛋白酶-2(Matrix metalloproteinase-2, MMP-2)的表达和临床意义。方法 采用免疫组织化学方法检测40例人胃癌组织、人胃息肉组织和40例正常胃黏膜组织中音猬因子、胶质瘤相关癌基因同源物-1、金属基质蛋白酶-2蛋白的表达。结果 胃癌组织中音猬因子、胶质瘤相关癌基因同源物-1、金属基质蛋白酶-2的阳性表达率分别为62.5%、67.5%、72.5%,高于胃息肉组织(阳性表达率分别为27.5%、37.5%、32.5%)和正常胃黏膜组织(阳性表达率分别为22.5%、17.5%、12.5%)(P<0.05);以上三者的表达与患者性别、年龄、组织学类型无关(P>0.05);而与分化程度、浸润深度、淋巴结转移相关(P<0.05);音猬因子、胶质瘤相关癌基因同源物-1、金属基质蛋白酶-2表达呈正相关。结论 刺猬信号通路可能通过某些机制可上调金属基质蛋白酶-2的表达,从而增强胃癌的侵袭性。联合检测胃癌组织中音猬因子、胶质瘤相关癌基因同源物-1、金属基质蛋白酶-2的表达水平在一定程度上可以作为胃癌预后的客观参考指标。

Objective To investigate the expression and clinical significance of Sonic hedgehog(Shh), Glioma-associated oncogene homolog -1(Gli-1) and Matrix metalloproteinase-2(MMP-2) in gastric cancer.Shh and Gli-1 are the molecules of Hedgehog(Hh) signaling pathway. MMP-2 is the member of matrix metalloproteinase family. Methods The expression of Shh,Gli-1 and MMP-2 proteins was examined by immunohistochemistry in the human gastric cancer tissues and the human gastric polyp and the normal gastric mucosa tissues of 40 cases. Results The positive expression rates of Shh,Gli-1 and MMP-2 in gastric cancer were 62.5%,67.5% and 72.5% respectively, which were significantly higher than those in the gastric polyp tissues (the positive expression rates were 27.5%,37.5% and 32.5% respectively) and normal gastric mucosa tissues (the positive expression rates were 22.5%,17.5% and 12.5% respectively),P<0.05.The expression of Shh,Gli-1 and MMP-2 was not correlated with the sex,age or histological type(P>0.05),but was correlated with depth of invasion,differentiation level and lymphonode metastasis in gastric cancer(P<0.05). The expression of Shh and Gli-1 was positive correlated with MMP-2. Conclusion Hedgehog(Hh)signaling pathway may have great effects on enhancing the invasive ability of gastric cancer by upregulating MMP-2 protein through some unknown mechanisms.The combined detection of the expression level of Shh,Gli-1 and MMP-2 in gastric cancer tissues might be used as an Objective references for assessing the prognosis of gastric cancer.

目的 对乳腺癌中血管内皮生长因子－C(VEGF－C)的表达与淋巴结转移及预后的关系展开研究分析。方法 随机选取我院接收救治的50例乳腺癌患者,采用免疫组化法检测50例患者乳腺癌中VEGF－C的表达情况,研究乳腺癌VEGF－C的表达与淋巴结转移及预后的关系。结果 50例乳腺癌患者中,淋巴结节转移组,VEGF－C阳性23例,阳性率92.0%;未见淋巴结节转移组,VEGF－C阳性10例,阳性率40.0%;淋巴结节转移组VEGF－C阳性表达率高于未见淋巴结节转移组;不同年龄、肿瘤直径以及病理分型的乳腺癌,VEGF－C阳性表达率差异无统计学意义(P均＞0.05);不同临床分期乳腺癌中,I~II期乳腺癌VEGF－C阳性表达率(58.1%)低于III~IV期VEGF－C阳性表达率(84.2%),数据差异有统计学意义(P＜0.05)。结论 早期检测乳腺癌中VEGF－C表达情况,能够为临床早期判定乳腺癌是否转移提供一项可测参考指标,对临床治疗、预后评估可起到一定参考价值。

Objective To make expand research and analysis for breast cancer and vascular endothelial growthfactor-C(VEGF-C)expression and lymph node metastasis and prognosis. Methods 50 cases of breast cancer patients were random collected in our hospital to detect the expression of VEGF-C in patients with breast cancer using immunohistochemical staining,the relationship between breast cancer VEGF-C expression and lymph node metastasis and prognosis. Results In 50 cases of breast cancer,lymph node metastasis group,VEGF-C positive in 23 cases,the positive rate is 92.0%;no lymph node metastasis group,VEGF-C positive in 10 cases,the positive rate is 40.0%;lymph node metastasis group VEGF-C positive expression rate was significantly higher than that no lymph node metastasis group;different age,tumor size and histological type of breast cancer,the VEGF-C positive expression rate difference was not statistically significant(P＞0.05);different clinical stages of breast cancer,I ~ II breast cancer VEGF-C positive expression rate(58.1%)was significantly lower than the III ~ IV of VEGF-C positive expression rate(84.2%),the data were statistically significant differences(P<0.05). Conclusion Early detection of breast cancer in the expression of VEGF-C can determine for early clinical metastasis of breast cancer,can provide a reference index for clinical treatment and prognosis.

目的 观察鼻咽癌患者癌组织中RhoA蛋白-Rho激酶蛋白表达情况。方法 收集增城市人民医院2009年2月—2014年6月耳鼻喉科住院治疗,进行活检的鼻咽癌患者切除标本共62例,包括癌组织及癌旁组织。通过SABC免疫组织化学法检测患者癌组织及癌旁组织中RhoA蛋白-Rho激酶蛋白表达情况。结果 鼻咽癌组织RhoA蛋白阳性表达率及Rho激酶蛋白阳性率高于癌旁组织(P=0.00);鼻咽癌癌组织RhoA及Rho激酶表达在年龄及性别分布上无差异,而TNM分期中Ⅲ期+Ⅳ期RhoA及Rho激酶表达阳性率高于Ⅰ期+Ⅱ期患者,同时存在淋巴结转移患者其RhoA及Rho激酶表达阳性率高于未转移患者。结论 鼻咽癌患者癌组织中RhoA蛋白及Rho激酶表达阳性率高,TNM分期越高及存在淋巴结转移者其RhoA蛋白及Rho激酶表达阳性率越高。RhoA-Rho激酶信号通路参与了鼻咽癌发生发展的过程。

Objective To observe the expression of RhoA-Rho kinase protein in patients with nasopharyngeal carcinoma tissue. Methods Collecting 62 cases of patients with nasopharyngeal carcinoma who hospitalized and hadbiopsyof specimens, including carcinoma tissues and adjacent tissues in department of otolaryngology-head and neck surgery from February 2010 to October 2014. The expression of RhoA-Rho kinase protein in carcinoma tissues and adjacent tissues was detected with SABC immune histochemical method. Results Positive expression rate of nasophryngeal carcinoma tissue RhoA protein and Rho kinase protein was significantly higher than that of adjacent tissues, and there was no obvious difference in age and sex between the expression of nasopharyngeal carcinoma tissue RhoA and Rho kinase(P=0.00), while patients being in the third and fourth periods of TNM stage had a higher positive expression rate of RhoA and Rho kinase than those being in the first and second periods. At the same time, RhoA and Rho kinase positive expression rate in patients with lymph node metastasis was higher than that in those without lymph node metastasis. Conclusion Positive expression rate of RhoA protein and Rho kinase in patients with nasopharyngeal carcinoma. The higher TNM stage is, the higher positive expression rate of RhoA protein and Rho kinase in lymph node metastasis is. RhoA-Rho kinase signaling pathways involved in the occurrence and development of nasopharyngeal carcinoma.