血浆巨噬细胞刺激蛋白表达在AECOPD的临床研究

目的研究患者血浆巨噬细胞刺激蛋白(macrophage stimulating protein,MSP)表达在AECOPD的临床研究。方法随机选择我院收治的60例急性加重期COPD(AECOPD)患者,60例稳定期COPD患者,并选择年龄、性别配对的60例正常体检人员设为对照组。检测三组受试者血浆MSP水平以及白介素6(in-terleukin-6,IL-6)、IL-8等炎性因子水平,并进行MSP水平与血清炎性因子水平及肺动脉压的相关性分析。结果 AECOPD组血浆MSP水平显著高于稳定期COPD组与对照组,稳定期COPD组显著高于对照组,(P 0. 05); AECOPD组血清IL-6、IL-8、肿瘤坏死因子α(Tumor necrosis factorα,TNF-α)水平显著高于稳定期COPD组与对照组,稳定期COPD组显著高于对照组,(P 0. 05);血浆MSP表达水平与血清IL-6、IL-8、TNF-α水平、平均肺动脉压(mm Hg)呈显著正相关(P 0. 05)。结论 MSP与COPD有关,与急性加重有关,可能与MPS通过调控巨噬细胞参与COPD气道炎症反应有关,其表达水平变化有助于COPD诊断与治疗指导。     

巨噬细胞刺激蛋白对烟熏大鼠肺泡巨噬细胞氧化应激和细胞因子产生的影响

目的 探讨巨噬细胞刺激蛋白(MSP)对烟熏大鼠肺泡巨噬细胞氧化应激和细胞因子产生的影响.方法 培养正常和烟熏不同时间(1个月、2个月、3个月)的大鼠肺泡巨噬细胞,给予不同浓度MSP处理24 h,采用酶联免疫法检测细胞上清液中细胞因子肿瘤坏死因子α(TNF-α)、白介索8(IL-8)和IL-1β的浓度,比色法检测细胞上清液中丙二醛(MDA)和超氧化物歧化酶(SOD)的水平.结果 ①MSP呈浓度依赖性促进正常组和各烟熏组大鼠肺泡巨噬细胞分泌TNF-α、IL-8和IL-1β;经MSP处理后,各烟熏组大鼠肺泡巨噬细胞上清液中TNF-α、IL-8和n-1β浓度均高于正常组(P＜0.05);大鼠肺泡巨噬细胞上清液中TNF-α、IL-8和IL-1β浓度随烟熏时间延长呈时间依赖性增加.②MSP呈浓度依赖性促进正常组和各烟熏组大鼠肺泡巨噬细胞分泌MDA,抑制其产生SOD;烟熏2个月组和烟熏3个月组大鼠肺泡巨噬细胞上清液中MDA水平均高于正常组(P＜0.05),SOD水平均低于正常组(P＜0.05);随着烟熏时间延长,大鼠肺泡巨噬细胞上清液中MDA水平呈时间依赖性增加,SOD水平呈时间依赖性降低.结论 MSP呈浓度依赖性促进正常和烟熏大鼠肺泡巨噬细胞分泌TNF-α、IL-8、IL-1β和MDA,抑制其产生SOD.MSP促烟熏大鼠肺泡巨噬细胞分泌TNF-α、IL-8、IL-1β、MDA及抑制其产生SOD的作用较正常大鼠更显著,且烟熏时间越长此作用越明显.     

RNA干扰技术阻断小鼠巨噬细胞受体相互作用蛋白2表达的实验研究

目的观察阻断受体相互作用蛋白2（Rip2）对巨噬细胞产生炎症细胞因子的影响,以及对内毒素血症小鼠的保护作用。方法构建Rip2小干扰RNA（siRNh）重组表达质粒,转染细胞后RT．PCR和Western blot检测Rip2的mRNA和蛋白表达,四甲基偶氮唑盐（M1Tr）法检测细胞增殖水平,脂多糖（LPS）刺激后,测定TNFa和高迁移率组蛋白1（HMGB1）的水平。Rip2 siRNA质粒转染小鼠后,观察小鼠病死率,测定血清TNFct水平和肝组织Rip2和HMGB1表达。结果Rip2 siRNA表达质粒可阻断Rip2 mRNA和蛋白表达。Rip2阻断的细胞增殖明显,LPS刺激后产生TNFα、HMGB1减少;Rip2阻断的小鼠生存率较其他组高（P〈0．05）,肝组织中HMGB1[（40．21±11．03）Pg／g]表达和血清TNFα[（300．43±59．26）ng／L]水平均较其他组低（P〈0．05）。结论Rip2 siRNA表达质粒可阻断Rip2的表达,从而减少TNFα、HMGB1等炎症细胞因子的产生,降低小鼠内毒素血症的病死率。     

巨噬细胞集落刺激因子对小鼠腹腔巨噬细胞清道夫受体途径和细胞内胆固醇酯积聚的影响

为探索巨噬细胞集落刺激因子、清道夫受体、氧化型低密度脂蛋白与动脉粥样硬化的关系，观察了重组人巨噬细胞集落刺激因子对小鼠腹腔巨噬细胞清道夫受体途径的影响以及重组人巨噬细胞集落刺激因子对氧化型低密度脂蛋白所致细胞内胆固醇酯积聚的影响．结果发现重组人巨噬细胞集落刺激因子能增加培养的小鼠腹腔巨噬细胞表面的清道夫受体数目，使之对氧化型低密度脂蛋白的结合和降解呈现剂量和时间依赖性增加，并使细胞内胆固醇酯积聚增多．表明巨噬细胞集落刺激因子可通过增加清道夫受体数目使小鼠腹腔巨噬细胞对氧化型低密度脂蛋白的结合和降解增多，从而增加细胞内胆固醇酯含量，促进动脉壁内泡沫细胞形成．     

胃癌的浸润、转移与E—钙粘附素—粘连素系统失活

E－钙粘附素－粘连素系统是上皮细胞间紧密连接的分子基础,其功能失活与源于上皮的癌肿的浸润与转移密切相关。本文综述该系统失活在胃癌发生发展中的作用。     

粒-巨噬细胞集落刺激因子在肺泡蛋白沉积症患者中的表达

目的 研究粒－巨噬细胞集落刺激因子（GM－CSF）蛋白及mRNA在4例肺泡蛋白沉积症（PAP）患者中的表达,探索PAP的发病机制。方法 采用ELISA法测定GM－CSF蛋白,采用RT－PCR检测外周血单个核细胞及肺泡巨噬细胞中GM－CSFmRNA表达水平。GM－CSFcDNA测序采用双脱氧链终止反应法。结果 4例PAP中3例外周血单个核细胞和肺泡巨噬细胞均检测不到GM－CSF释放,但mRNA表达水平均正常。cDNA测序发现1例PAP患者GM－CSFcDNA第382位碱基发生点突变（T→C）,致117位氨基酸序列发生改变（异亮氨酸→苏氨酸）。结论 GM－CSF蛋白表达异常与PAP发病可能有关,GM－CSFcDNA点突变可能是导致GM－CSF蛋白表达异常的原因之一。     

尿巨噬细胞趋化蛋白-1和巨噬细胞集落刺激因子对狼疮肾炎复发的诊断作用

目的探讨在成功的泼尼松和环磷酰胺诱导治疗后,引起弥漫增生型(Ⅳ型)狼疮肾炎复发的预测指标。方法收集弥漫增生型狼疮肾炎病例,将泼尼松和环磷酰胺诱导治疗成功的病例纳入研究对象。记录临床和实验室资料,于治疗开始后6个月时检测患者尿巨噬细胞趋化蛋白(MCP)-1和巨噬细胞集落刺激因子(M-CSF)。追踪治疗后的复发情况。结果共收集到64例诱导治疗成功的病例,经平均(27±3)个月随访,18例(28%)患者发生至少一次肾性复发,其复发的平均时间为(14±4)个月。复发组患者尿MCP-1和M-CSF水平明显高于缓解维持组。尿MCP-1和M-CSF升高,及血C3降低和抗dsDNA抗体阳性均是Ⅳ型狼疮肾炎复发的独立预测因子。有7例患者出现血肌酐倍增(CRX2),肾性复发是CRX2的惟一预测因子。结论尿MCP-1和M-CSF持续升高是Ⅳ型狼疮肾炎复发的独立预测因子。该研究提示监测诱导缓解患者肾组织炎症指标可有利于指导狼疮肾炎的治疗。     

粒-巨噬细胞集落刺激因子及肺表面活性蛋白在肺泡蛋白沉积症患者肺内的表达

目的通过观察成人特发性肺泡蛋白沉积症(PAP)患者肺部粒-巨噬细胞集落刺激因子(GM-CSF)及其受体蛋白以及肺泡表面活性蛋白的表达,探讨PAP的发病机制。方法用SP免疫组化技术检测6例PAP患者(PAP组)及6例对照者(对照组)肺组织肺泡巨噬细胞及Ⅱ型肺泡上皮细胞GM-CSF、GM-CSF/IL-3/IL-5共同β链受体(GM-CSFβcR)、肺表面活性蛋白(SP)-A和SP-D蛋白表达。结果(1)肺泡巨噬细胞2组表达GM-CSF蛋白均较弱,阳性细胞数少,组间比较差异无统计学意义(P=0.818);2组均表达GM-CSFβcR蛋白,对照组显著高于PAP组(P=0.002);2组均表达SP-A蛋白,且PAP组呈阳性反应,但阳性细胞数少,积分指数显著低于对照组(P=0.004);2组表达SP-D蛋白差异无统计学意义(P=0.24)。(2)Ⅱ型肺泡上皮细胞2组均表达SP-A和SP-D蛋白,组间比较差异无统计学意义(P=0.818,P=0.485);对照组少数表达GM-CSFβcR蛋白,PAP组无GM-CSFβcR蛋白表达;2组均无GM-CSF蛋白表达。结论肺泡巨噬细胞表达GM-CSFβcR蛋白减少可能与成人特发性PAP肺泡巨噬细胞功能障碍有关。     

糖基化终产物对U937巨噬细胞高密度脂蛋白受体蛋白表达的影响

目的　研究诱导分化及糖基化终产物 (AGEs)对人单核 /巨噬细胞 (U937细胞 )高密度脂蛋白受体 (SR BI)蛋白表达的影响 ,探讨AGEs和巨噬细胞SR BI在动脉粥样硬化中的作用。方法　U937细胞经PMA诱导分化 ,并将不同浓度或同一浓度AGEs与诱导分化 48h后的U937细胞共同孵育 ,用免疫细胞化学法和Western印迹法检测SR BI蛋白的表达。结果　诱导分化后U937细胞SR BI表达在 2 4、48h逐渐升高 ,72h下降 ;1 0 0、2 0 0和 40 0 μg/mlAGEs刺激后细胞表面SR BI蛋白表达量分别是BSA组的 1 44、2 38和 2 77倍 (P <0 0 5) ;40 0 μg/ml的AGEs作用 6、1 2、2 4、48h后 ,U937巨噬细胞SR BI蛋白表达量分别为 0h的 1 38、2 49、3 76和 4 2 5倍 (P <0 0 5)。结论　AGEs可增加U937巨噬细胞SR BI蛋白表达且呈浓度和时间依赖性。     

Death decoy receptor overexpression and increased malignancy risk in colorectal cancer

AIM:To evaluate human epidermal growth factor receptor 2(HER2)and death decoy receptor(DcR3)as colorectal cancer prognostic indicators.METHODS:Colorectal carcinoma specimens from 300patients were analyzed by immunohistochemistry to detect the staining patterns of HER2 and DcR3.Classification of HER2 staining was carried out using the United States Food and Drug Administration semi-quantitative scoring system,with scores of 0 or 1+indicating a tumor-negative(normal expression)status and scores of 2+and 3+indicating a tumor-positive(overexpression)status.Classification of DcR3 was carried out by quantitating the percentage of positive cells within the stained section,with<10%indicating a tumor-negative status and≥10%indicating a tumor-positive status.Correlation of the HER2 and DcR3 staining status with clinicopathological parameters[age,sex,tumor size,differentiation,and the tumor,node,metastasis(pTNM)classification]and survival was statistically assessed.RESULTS:Tumor-positive status for HER2 and DcR3was found in 18.33%and 58.33%of the 300 colorectal carcinoma specimens,respectively.HER2 tumorpositive status showed a significant correlation with tumor size(P=0.003)but not with other clinicopathological parameters.DcR3 tumor-positive status showed a significant correlation with tumor differentiation(P<0.001),pTNM stage(P<0.001),and lymph node metastasis(P<0.001).However,correlation coefficient analysis did not indicate that a statistically significant correlation exists between tumor-positive status for the HER2 and DcR3 overexpression(P=0.236).Patients with specimens classified as DcR3-overexpressing had a significantly worse overall survival(OS)rate than those without DcR3 overexpression(median OS:42.11vs 61.21 mo;HR=50.27,95%CI:44.90-55.64,P<0.001).HER2 overexpression had no significant impact on median OS(35.10 mo vs 45.25 mo;HR=44.40,95%CI:39.32-49.48,P=0.344).However,patients with specimens classified as both HER2-and DcR3-overexpressing had a significantly poorer median OS than those with only HER2 overexpression(31.80 mo vs 52.20 mo;HR=35.10,95%CI:22.04-48.16,P=0.006).CONCLUSION:HER2 overexpression is not an independent prognostic marker of colorectal cancer,but DcR3 overexpression is highly correlated with lymph node metastasis and poor OS.     

Tyrosine kinase of insulin-like growth factor receptor as target for novel treatment and prevention strategies of colorectal cancer

INTRODUCTION Colorectal cancer (CRC) is the third most common cancer and the fourth most frequent cause of cancer-related deaths worldwide. Long-term survival of colorectal cancer is related to the stage of disease. Once distal metastases develop the prog…     

Blocking effects of siRNA on VEGF expression in human colorectal cancer cells

AIM:To investigate the expression of vascular endothelial cell growth factor (VEGF) and its receptors Fmslike tyrosine kinase 1 (FLT-1) and fetal liver kinase 1 (FLK-1) in colorectal carcinoma (CRC),and the blocking effects of small interfering RNAs (siRNAs) on VEGF expression in human colorectal cancer HCT116 cells.METHODS:Immunohistochemical staining for VEGF,FLT-1 and FLK-1 proteins was performed in 82 cases of CRC and 14 normal colorectal mucosae.A siRNA targeting VEGF was synthesized and transfected into HCT116 cells using lipofectamine 2000.Immunocytochemical staining and Western blotting analyses were performed to detect the expression of VEGF protein.The suppressive effect of the siRNA on cell proliferation was detected using the 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltertrazolium bromide (MTT) assay.Cellular apoptosis was detected using flow cytometry (FCM).RESULTS:The expression of VEGF,FLT-1 and FLK-1 in tumor tissues was significantly higher than that in normal tissues (P=0.008,P=0.000,P=0.000).The expression of VEGF was positively correlated with both lymph node metastasis and clinical stage (P=0.009 and P=0.025,respectively).Immunocytochemistry showed that the expression of VEGF was weakly positive and Western blotting indicated a significant reduction in VEGF-siRNA cell protein levels.VEGF-siRNA cell growth inhibition was assessed by the MTT assay,and the tumor cell proliferation rate was significantly different at 24,48,and 72 h after transfection.FCM results showed that the VEGF-siRNA group had an apparent aneuploid peak.CONCLUSION:VEGF,FLT-1 and FLK-1 are associated with colorectal carcinogenesis.siRNA silencing of the VEGF gene suppresses proliferation,and induces apoptosis in HCT116 cells.The results suggest that VEGF may be a new gene therapy target for colorectal cancer.     

巨噬细胞集落刺激因子对破骨细胞活性的影响

巨噬细胞集落刺激因子(M-CSF)是目前临床上应用最广泛的细胞因子之一,因其具有独特和良好的升白细胞及干细胞动员作用,得到临床医师的肯定。M-CSF可诱导人破骨细胞形成,刺激其分化、存活,参与成熟破骨细胞对骨的吸收作用;而且可以抑制破骨细胞凋亡,因而可用于治疗骨骼石化症。虽然有关M-CSF对破骨细胞的生长、分化和凋亡的影响已有较广泛的研究,但对其确切的细胞生物学机制并不很清楚,本文综述了近年来该方面的研究进展。     

促甲状腺激素受体胞内环基因突变与乳头状甲状腺癌发病的相关性研究

目的探讨乳头状甲状腺癌的发病与促甲状腺激素受体(TSHR)基因突变的相关性。方法采用巢氏-多聚酶联反应-单链构象多态性分析(NEST-PCR-SSCP)和DNA测序方法,对65例乳头状甲状腺癌和44例正常甲状腺组织TSHR3个胞内环基因进行检测。结果NEST-PCR-SSCP检测乳头状甲状腺癌促甲状腺激素受体(TSHR)3个胞内环未发现明显带型异常;DNA测序后,检测的第3胞内环2例对照组织和3例甲状腺癌组织TSHR2000位点碱基均由C→T,使得所编码的601位氨基酸由组氨酸(CAT)→酪氨酸(TAT)(His→Tyr),余胞内环未发现基因突变。结论乳头状甲状腺癌TSHR3个胞内环未发现基因突变,提示乳头状甲状腺癌发病与TSHR胞内环基因突变无关;5例标本601位氨基酸均为酪氨酸,考虑中国人TSHR基因可能与国外人群存在差异,TSHR2000位点碱基存在基因多态性。     

Anticancer effects of sweet potato protein on human colorectal cancer cells

AIM: To investigate the effects of proteins purified from sweet potato storage roots on human colorectal cancer cell lines. METHODS: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Hoechst 33258 nuclear staining and Boyden transwell chamber methods were used to determine whether purified sweet potato protein (SPP) from fresh sweet potato roots affected proliferation, migration and invasion, respectively, of human colorectal cancer SW480 cells in vitro . The inhibitory effects of SPP on growth of human colorectal cancer HCT-8 cells intraperitoneally xenografted in nude mice and spontaneous lung metastasis of murine Lewis lung carcinoma 3LL cells subcutaneously transplanted in C57 BL/6 mice were also investigated in vivo . RESULTS: SPP inhibited the proliferation of SW480 cells in a dose-dependent manner, with an IC 50 value of 38.732 μmol/L (r2 = 0.980, P = 0.003) in the MTT assay. Hoechst 33258 nuclear staining further revealed inhibition of cell viability and induction of apoptosis by SPP. The transwell assay disclosed significant reduction in migrated cells/field by 8 μmol/L SPP (8.4 ± 2.6 vs 23.3 ± 5.4, P = 0.031) and invaded cells/field through the ECMatrix by 0.8 μmol/L SPP, compared with the control (25.2 ± 5.2 vs 34.8 ± 6.1, P = 0.038). Both intraperitoneal (ip) and intragastric (ig) administration of SPP led to significant suppression of growth of intraperitoneally inoculated HCT-8 cells in nude mice to 58.0% ± 5.9% (P = 0.037) and 43.5% ± 7.1% (P = 0.004) of the controls, respectively, after 9 d treatment. Bloody ascites additionally disappeared after ip injection of trypsin inhibitor. Notably, ig and ip administration of SPP induced a significant decrease in spontaneous pulmonary metastatic nodule formation in C57 BL/6 mice (21.0 ± 12.3 and 27.3 ± 12.7 nodules/lung vs 42.5 ± 4.5 nodules/lung in controls, respectively, P < 0.05) after 25 d treatment. Moreover, the average weight of primary tumor nodules in the hind leg of mice decreased from 8.2 ± 1.3 g/mice in     

Size does not determine the grade of malignancy of early invasive colorectal cancer

AIM： To clarify the clinicopathological characteristics of small and large early invasive colorectal cancers （EI-CRCs）, and to determine whether malignancy grade depends on size. METHODS： A total of 583 consecutive EI-CRCs treated by endoscopic mucosal resection or surgery at the National Cancer Center Hospital between 1980 and 2004 were enrolled in this study. Lesions were classified into two groups based on size： small （≤10 ram） and large （〉10 ram）. Clinicopathological features, incidence of lymph node metastasis （LNM） and risk factors for LNM, such as depth of invasion, lymphovascular invasion （LVI） and poorly differentiated adenocarcinoma （PDA） were analyzed in all resected specimens. RESULTS： There were 120 （21%） small and 463 （79%） large lesions. Histopathological analysis of the small lesion group revealed submucosal deep cancer （sin： 1〉1000 μm） in 90 （75%） cases, LVI in 26 （22%） cases, and PDA in 12 （10%） cases. Similarly, the large lesion group exhibited submucosal deep cancer in 380 （82%） cases, LVI in 125 （27%） cases, and PDA in 79 （17%） cases. The rate of LNM was 11.2% and 12.1% in the small and large lesion groups, respectively.CONCLUSION： Small EI-CRC demonstrated the same aggressiveness and malignant potential as large cancer.     

Stromal expression of urokinase-type plasminogen activator receptor (uPAR) is associated with invasive growth in primary liver cancer

Abstract: Aims/Background: Expression of urokinase-type plasminogen activator receptor (uPAR) was studied in 25 hepatocellular carcinomas (HCCs) and seven cholangiocellular carcinomas (CCCs) by immunohistochemistry. Methods and Results: uPAR was expressed mostly by host cells distributed along the tumour-host interface in all cases of HCC and CCC, and its expression was higher in CCC. These uPAR-positive cells were identified as macrophages by observation of serial sections stained for CD68, a marker for macrophages. Cancer cells were positive for uPAR in only one case of poorly differentiated HCC with sarcomatous changes and in three cases of CCC. Hepatocellular carcinomas were classified into two types: those with a fibrous capsule (expansive type) and those without a fibrous capsule (invasive type). Invasive-type HCCs showed more prominent expression of uPAR by macrophages than expansive HCCs (p<0.001), to approximately the same degree as that of CCC. Extrahepatic metastasis was observed in two of 16 expansive HCCs, five of nine invasive HCCs and six of seven CCCs. Conclusions: Our findings suggest that uPAR expression mainly by macrophages is associated with invasive growth of cancer cells into the surrounding tissue in primary carcinoma of the liver.