三磷酸腺苷结合盒转运体成员2在人脑胶质瘤中的表达及其与神经巢蛋白表达的关系

目的 探讨三磷酸腺苷结合盒转运体成员2(ABCG2)在人脑胶质瘤组织中的表达及其与胶质瘤分级和神经巢蛋白(nestin)表达的关系.方法 实时荧光定量聚合酶链反应(PCR)检测ABCG2在52例不同病理级别胶质瘤中的mRNA表达;免疫组织化学SABC法检测ABCG2和nestin的蛋白表达.结果 ABCG2在脑胶质瘤中的mRNA表达与正常脑组织比较呈过表达(P<0.05),且随着病理级别的升高而增加(P<0.05);免疫组织化学显示ABCG2在52例胶质瘤中的阳性表达率为32.7%(17/52),并与病理级别明显相关(x2=4.62,P<0.05),主要呈亲血管分布;ABCG2的表达与nestin的表达明显相关(x2=7.60,P<0.05).结论 ABCG2在人脑胶质瘤中的表达随着病理级别的升高而逐渐增加,且与nestin的表达呈正相关;脑肿瘤干细胞可能与神经干细胞具有一定的同源性.     

特异性抑制髓母细胞瘤1型胰岛素生长因子受体信号通路后的差异性基因表达

目的 应用聚合酶链反应(PCR)芯片技术观察特异性抑制髓母细胞瘤细胞1型胰岛素生长因子受体(IGF-1R)信号通路的差异性基因表达,探讨IGF-1R在髓母细胞瘤细胞增殖中的作用.方法 体外培养髓母细胞瘤Doay细胞株,使用不同终质量浓度NVP-ADW742作用48 h和24 h后,检测细胞增殖和凋亡,分别使用IGF-1R酪氨酸蛋白激酶抑制剂NVP-ADW742终质量浓度为0、0.5、1.0、2.0、5.0、10.0、20.0、50.0μmol/L作用48 h后检测细胞存活率,以及终质量浓度为0.1、1.0、2.0、5.0、10.0μmol/L作用24 h后检测凋亡率,应用PCR芯片技术检测IGF-1R信号通路的差异性基因表达,并采用Western blot检测其蛋白表达.结果 在对应浓度的NVP-ADW742作用下Doay细胞的存活率分别为95.95%、93.95%、91.97%、86.60%、70.57%、15.01%、1.63%(P＜0.05),凋亡率分别为6.92%、8.83%、11.22%、17.27%、29.98%(P＜0.05),两者呈剂量依赖性,其浓度越高,抑制和促凋亡作用越强;在PCR芯片检测有14个差异表达基因;Western blot检测表明在2μmol/L的NVP-ADW742作用下,随时间延长PI3K、Akt、p38、GSK-3β和bcl-2蛋白表达逐渐下调.结论 髓母细胞瘤中IGF-1R信号通路异常可以促进肿瘤细胞的增殖,利用功能性PCR芯片可以有效地检测肿瘤细胞中相关基因在肿瘤增殖和凋亡中的作用.     

合肥地区小儿肺炎链球菌感染及菌型研究

本文报告小儿肺炎链球菌感染34例,以菌血症、肺炎及脑膜炎为主,有32例,占94.1％。34例获菌株39株,除4株血与脑脊液培养重复者外,35株可分成11个血清型(群),占前四位的有6个型(群):5型、1型、6群、2型、14型及12群,共29株,占82.9％,此为今后制造疫苗提供了科学依据。     

三磷酸腺苷结合盒转运体成员2在人脑胶质瘤中的表达及其与神经巢蛋白表达的关系

Objective To study the expression of ATP-binding cassette superfamily G member 2 ( ABCG2) and its relationship with the malignant degree and expression of nestin in human gliomas. Methods Fifty-two gliomas of different WHO grades and 8 normal brain tissues were detected for the mRNA expression of ABCG2 by using real-time quantitive polymerase chain reaction (PCR). And the protein expression of ABCG2 and nestin was detected by immunohistochemistry. Results The mRNA of ABCG2 was overexpressed in gliomas as compared with that in normal brain tissues (P<0. 05), and up-regulated with the increase of pathologic degrees (P<0. 05 ). The positive protein expression rate of ABCG2 in 52 gliomas was 32.7%. The positive rate was significantly higher in grade Ⅲ-Ⅳ than in grade Ⅰ -Ⅱ (x2 =4. 62, P < 0. 05). And the expression level of ABCG2 was obviously related with the expression of nestin (x2= 7. 60,P < 0.05). Conclusion The expression level of ABCG2 was obviously related with glioma pathological grade and the expression of nestin. Brain tumor stem cells may have some homology with nerve stem cells.     

三磷酸腺苷结合盒转运体成员2在人脑胶质瘤中的表达及其与神经巢蛋白表达的关系

Objective To study the expression of ATP-binding cassette superfamily G member 2 ( ABCG2) and its relationship with the malignant degree and expression of nestin in human gliomas. Methods Fifty-two gliomas of different WHO grades and 8 normal brain tissues were detected for the mRNA expression of ABCG2 by using real-time quantitive polymerase chain reaction (PCR). And the protein expression of ABCG2 and nestin was detected by immunohistochemistry. Results The mRNA of ABCG2 was overexpressed in gliomas as compared with that in normal brain tissues (P<0. 05), and up-regulated with the increase of pathologic degrees (P<0. 05 ). The positive protein expression rate of ABCG2 in 52 gliomas was 32.7%. The positive rate was significantly higher in grade Ⅲ-Ⅳ than in grade Ⅰ -Ⅱ (x2 =4. 62, P < 0. 05). And the expression level of ABCG2 was obviously related with the expression of nestin (x2= 7. 60,P < 0.05). Conclusion The expression level of ABCG2 was obviously related with glioma pathological grade and the expression of nestin. Brain tumor stem cells may have some homology with nerve stem cells.     

大黄酸通过抑制ERK途径诱导胶质瘤细胞增殖分化的实验研究

目的探讨大黄酸对胶质瘤细胞增殖和分化的影响,以及其可能的机制。方法大鼠F98和人Hs683细胞系分别予以不同浓度的大黄酸处理(大黄酸处理组)和单用培养基培养(对照组)。采用四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)比色法检测各组细胞的增殖能力;倒置显微镜观察细胞形态变化;免疫细胞化学检测细胞胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)的表达。ELISA法检测经25μmol/L大黄酸处理不同时间的细胞磷酸化细胞外信号调节激酶(phospho-excelluar signal-regulated kinase,p-ERK)的活性。结果与对照组相比,大黄酸处理组F98细胞的相对活性降低,细胞胞体失去了对照组细胞的多边形形状,胞体变小,并发出较长的突起;免疫细胞化学检测发现GFAP表达增加。大黄酸处理后的F98细胞p-ERK相对活性降低。用ERK1/2特异性抑制剂PD98059处理的F98细胞及25μmol/L大黄酸处理的人Hs683细胞增值活性下降,细胞形态出现与大黄酸处理F98细胞相似的变化,GFAP表达亦明显增加。结论大黄酸抑制胶质瘤细胞增殖,并可能通过抑制ERK途径促进胶质瘤细胞向成熟星形胶质细胞分化。     

53例小儿细菌性脑膜炎有关病原研究

研究目的 小儿细菌性脑膜炎(菌脑)的病原构成及病原检测方法的阳性率比较。 研究方法 共53例,应用肺炎链球菌多价型(PNC)、流感嗜血杆菌B型(Hib)、脑膜炎双球菌A及B群(MCA,MCB)三种细菌抗血清,采用对流免疫电泳(CIE)检测脑脊液(CSF)、血、尿抗原共47例,CSF涂片革兰氏染色找细菌25例,CSF培养20例,血培养20例。 结果和结论 1.明确病原菌者86.7％,其中Hib占45.28％为主要病原菌,而MC28.3％占第二位,PNC占9.43％。2.检测方法中证明CIE检测抗原阳性率(89.4％)明显优于常规检查细菌的方法(15～35％),(P<0.01)。3.入院时第一次CSF,用CIE法检测病原阳性率最高。     

BMP受体在胶质母细胞瘤起始细胞早期分化中的表达

目的在分离鉴定胶质母细胞瘤U87细胞中的胶质瘤起始细胞(GTICs)的基础上,检测在GTICs细胞中骨形态发生蛋白(BMP)受体1A、B的表达,探讨BMP受体1A、B对GTICs所起的生物学作用。方法利用流式细胞术检测U87细胞中CD133和巢蛋白阳性细胞,细胞免疫荧光和蛋白印迹法检测GTICs和普通细胞中BMP受体1A、B的表达。结果 CD133和巢蛋白阳性表达的细胞在瘤U87细胞中所占的比例分别0.92%和5.72%;GTICs可诱导分化为表达β-微导管蛋白和胶质纤维酸性蛋白的细胞;U87细胞的GTICs中BMP受体1A表达明显增高,而经诱导分化培养后的BMP受体1A的表达又明显降低,BMP受体1B表达变化不明显。结论 BMPs信号通路中BMP受体对调控GTICs的增殖分化可能起着重要作用。     

PPM1D silencing by lentiviral-mediated RNA interference inhibits proliferation and invasion of human glioma cells

To construct a lentiviral shRNA vector targeting human protein phosphatase 1D magnesium-dependent(PPM1D) gene and detect its effectiveness of gene silencing in human gliomas,specific siRNA targets with short hairpin frame were designed and synthesized.DNA oligo was cloned into the pFU-GW-iRNA lentiviral expression vector,and then PCR and sequencing analyses were conducted to verify the constructs.After the verified plasmids were transfected into 293T cells,the lentivirus was produced and the titer of virus was determined.Real-time quantitative PCR and Western blot were performed to detect the PPM1D expression level in the infected glioma cells.PCR and Western blot analyses revealed the optimal interfering target,and the virus with a titer of 6×108 TU/mL was successfully packaged.The PPM1D expression in human glioma cells was knocked down at both mRNA and protein levels by virus infection.The expression of PPM1D mRNA and protein was decreased by 76.3% and 87.0% respectively as compared with control group.The multiple functions of human glioma cells after PPM1D RNA interference were detected by flow cytometry and cell counting kit-8(CCK-8).Efficient down-regulation of PPM1D resulted in significantly increased cell apoptosis and reduced cell proliferation and invasion potential in U87-MG cells.We have successfully constructed the lentiviral shRNA expression vector capable of stable PPM1D gene silencing at both mRNA and protein levels in glioma cells.And our data gave evidence that the reduced cell growth observed after PPM1D silencing in glioma cells was at least partly due to increased apoptotic cell death.