间日疟原虫深圳株红内期SSUrDNA基因片段的克隆与序列分析

目的 体外扩增间日疟原虫深圳株红内期小亚单位核糖体核糖核酸编码基因(SSUrDNA)片段，研究其结构与功能。方法 设计一对特异性引物，采用聚合酶链反应(PCR)从间日疟原虫患者血样中扩增出间日疟原虫SSUrDNA片段，以PUC19质粒T载体构建重组子导入大肠杆菌JM109；阳性克隆双酶切鉴定后，双脱氧末端终止法测定序列。结果 间日疟原虫SSUrDNA扩增片段大小为341bp；阳性克隆双酶切及PCR扩增均得到预期大小的片段；序列测定插入片段为341bp，与Sal I株顺序相比，仅在第151位处缺失一个碱基C。结论 成功克隆了间日疟原虫SSUrDNA片段．该序列在间日疟原虫虫株间高度保守。     

益气活血方对气虚血瘀证脑缺血再灌注损伤大鼠Fas、FasL蛋白表达的影响

目的：探讨益气活血方对气虚血瘀证局灶性脑缺血再灌注损伤大鼠Fas、FasL蛋白表达的影响。方法：采用气虚血瘀证局灶性脑缺血再灌注损伤大鼠模型，用线栓法阻塞大鼠大脑中动脉复制局部脑缺血再灌注模型，缺血2h后再灌注l，3，7d。用免疫组化染色法分别检测缺血皮质Fas、FasL蛋白表达。结果：与模型组比较。益气活血方组Fas、FasL蛋白表达显降低。结论：益气活血方可能通过抑制Fas、FasL蛋白表达，减轻气虚血瘀证局灶性脑缺血再灌注损伤。     

凋亡调控基因Fas/APO-1、bcl-2在胆囊癌中的表达及与生物学特性的关系

目的　研究肿瘤凋亡调控基因Fas/APO 1、bcl 2在胆囊癌发生发展中的作用。方法　应用免疫组化法对 2 8例胆囊癌及其癌旁组织、2 0例胆囊息肉和腺瘤组织检测基因Fas/APO 1、bc1 2的表达 ,并分析它们与胆囊癌生物学特性的关系。结果　胆囊癌组织中bc1 2表达率与癌旁组织、息肉和腺瘤组织相比显著增高 ,而Fas/APO 1表达明显降低 (均P <0 .0 5 ) ;与肿瘤生物学特性的关系 ,bc1 2蛋白表达率在高、中分化组中明显低于低、未分化组 ,NevinⅠ、Ⅱ、Ⅲ期低于Ⅳ、Ⅴ期 ,而Fas/APO 1蛋白表达则相反 (均P <0 .0 5 )。结论　bc1 2为胆囊癌细胞凋亡抑制基因 ,而Fas/APO 1则为促进基因 ,两者在胆囊癌的发生发展中起重要作用     

微型染色体维持蛋白2和Fas相关磷酸酯酶1与乳腺癌增殖和凋亡的关系

目的 探讨微型染色体维持蛋白2(MCM2)和Fas相关磷酸酯酶(FAP-1)的表达与乳腺癌增殖和凋亡的关系.方法 采用SP法检测60例乳腺癌及15例乳腺良性疾病中MCM2及FAP-1蛋白的表达水平,10例正常乳腺组织为对照组.结果 10例正常乳腺上皮、15例乳腺良性疾病、60例乳腺癌MCM2的阳性标记指数(LI)分别为6.6±1.1、14.7±3.2、44.7±4.3,各组间差异有统计学意义(P＜0.01);MCM2的阳性表达在乳腺癌TNM分期、不同组织学分级(Ⅰ、Ⅱ、Ⅲ)及淋巴结转移之间差异有统计学意义,呈升高趋势(P＜0.05);MCM2的LI均高于Ki-67的LI,MCM2和Ki-67在乳腺癌中的表达呈平行关系(P＜0.01).FAP-1的阳性表达率与乳腺癌的临床分期、病理分级及淋巴结转移性明显相关(P＜0.05);FAP-1在乳腺癌组织中的表达均显著高于正常组织和良性肿瘤.结论 MCM2、FAP-1在乳腺癌组织中的表达与乳腺癌的发生发展有密切联系,MCM2可能是更理想的细胞增殖标记物,FAP-1可能具有促进乳腺癌细胞增殖的作用.     

酪氨酸激酶Syk在肝癌中的表达及与血管生成的关系

目的探讨酪氨酸激酶Syk在肝细胞性肝癌中的表达及对肿瘤微血管生成的影响。方法采用RT-PCR检测Syk mRNA在肝细胞性肝癌及癌旁正常组织中的表达，免疫组化SABC法检测标本中CD34的表达反映肿瘤的微血管密度（MVD）。结果24例癌旁正常组织中Syk mRNA均表达阳性，32例肝细胞性肝癌中Syk mRNA表达率为46.9％（15／32），其中低分化组阳性表达率23．1％（3／13），明显低于高分化组阳性表达率63．2％（12／19）（P〈0．05）。肿瘤微血管密度（MVD）检测：低分化组（Ⅲ级Ⅳ级为49．2±3.6，54±4.3），明显高于高分化组（Ⅰ级Ⅱ级为13．6±4．5，32.3±3．2）与正常组织（5．9±1．7），有显著统计学意义（P〈0．05）。Syk mRNA的表达与CD34的表达明显负相关（r=-0.97）。结论肝细胞性肝癌中Syk基因的缺失对癌组织血管的生成起重要的作用。     

uPA、uPAR和PTEN在膀胱移行细胞癌中的表达及其与肿瘤侵袭和转移的关系

目的研究膀胱移行细胞癌uPA、uPAR及抑癌基因PTEN蛋白的表达及其与肿瘤侵袭、转移的关系。方法应用免疫组织化学SP法检测60例膀胱移行细胞癌组织uPA、uPAR和PTEN蛋白的表达。结果uPA、uPAR和PTEN阳性表达率分别为50.00%、50.00%和41.67%。uPA和uPAR的高表达与膀胱移行细胞癌的分化程度、浸润和转移关系密切(P<0.05);PTEN蛋白的低表达与膀胱移行细胞癌的分化程度、浸润和转移密切相关(P<0.05);膀胱癌中uPA和uP-AR与PTEN蛋白表达呈负相关(r=-0.481,P<0.05)。结论uPA和uPAR的高表达可能作为膀胱移行细胞癌侵袭、转移的指征之一,PTEN可能作为膀胱移行细胞癌预后不良的指标;uPA和uPAR对肿瘤细胞侵袭转移的作用可能在一定程度上受到PTEN的调控。     

Therapeutic effect of 5-FC on YCD modified murine P388 leukemia in vivo

Cytosine deaminases (CDs) in bacteria andfungi are found to deaminate prodrug 5 - FC intocytotoxic agent 5 - FU .Yeast CD (YCD) is clonedfrom Saccharomyces cerevisiae.It has been shownpreviously that YCD was more efficient inconversing5 - FC than bacterial CD(b CD) [1-3 ] .As anovel suicide gene therapy system,YCD/ 5 - Fcsystem may be promising in cancer therapy andprevention of graft versus host disease.In thepresent study,we established a P388/ DBA murineleukemia model by infusin…     

Determination of phospholipidosis potential based on gene expression analysis in HepG2 cells.

Phospholipidosis (PLD) is characterized by an intracellular accumulation of phospholipids in lysosomes and the concurrent development of concentric lamellar bodies. Recently, H. Sawada et al. (2005, Toxicol. Sci. 83, 282-292) identified 17 genes as potential biomarkers of PLD in HepG2 cells. The present study was undertaken to determine if this set of genes measured by quantitative PCR could be validated in the same cell line. The objective was also to investigate the dose-response relationship to further validate the assay and to select the concentrations to use for screening activities. In a first experiment (one concentration tested), out of the 17 genes, the best gene biomarkers of PLD (i.e., 11 genes) were selected for practical screening reasons. Based on these genes, 91.6% (i.e., 11 of 12) of the compounds known to induce PLD were identified as positive and all the negative compounds (i.e., five of five) were also confirmed. When the data obtained in the first experiment were compared to the data by Sawada et al., (2005) the coefficient of correlation calculated was slightly higher than 75%. In the second experiment (26 compounds [all 17 compounds from the first experiment plus 9 other compounds] tested at a minimum of three concentrations), 93.3% (14/15) of the compounds known to induce PLD were identified as such and all the negative controls (six compounds) were also confirmed. Three compounds likely to induce PLD were identified as positive in our assay. Finally, two compounds for which no data are available were also tested. When both experiments 1 and 2 were compared, the coefficient of correlation for 16 compounds tested at the same concentrations reached 87.7%. In conclusion, the present study further confirms the utility of gene expression in HepG2 cells to identify a potential to induce PLD. Finally, based on the data presented, researchers are encouraged to use a range of minimum three concentrations (e.g., 12.5, 25, and 50 microM) to screen for PLD in the human HepG2 cell line.     

超声引导下细针穿刺检测甲状腺乳头状癌 ret/PTC基因重组突变

目的了解ret／PTC基因突变与超声、病理诊断在甲状腺乳头状癌诊断中的关系。方法术前超声检查，并在超声引导下细针穿刺，活体取材进行病理诊断和逆转录聚合酶链（RT-PCR）方法检测ret／PTC基因突变。结果在34例超声与病理诊断的甲状腺乳头状癌（PTC）患者中，16例分子生物学检测发现ret／PTC基因突变（47％），其中8例为PTC-1基因突变（50％），2例PTC-2基因突变（12．5％），2例PCT-3基因突变（12．5％），3例PTC-1和PTC-2突变同时存在（18．75％），1例PTC-1和PTC-3突变同时存在（6．25％）。结论ret／PTC基因重组突变可存在于散发的甲状腺乳头状癌中，主要表现PTC-1型。应用超声引导下细针穿刺与甲状腺乳头状癌基因突变的检测相结合是早期诊断甲状腺乳头状癌的有效方法。     

FasL基因表达对缺氧状态下直肠癌细胞增殖凋亡的影响

Objective To elucidate the effect of FasL gene expression on the proliferation and apoptosis of hypoxic rectal carcinoma cells. Methods The normoxic expression level of FasL in HR-8348 subtype cells (HR-8348B, HR-8348L, HR-8348F and HR-8348As) with different invasive power were verified by Western blot. Hypoxia models for HR-8348B, HR-8348L, HR-8348F and HR-8348As were constructed with chemical modeling, then the FasL levels in all groups at 12 h after hypoxia were quantitated by Western blot. Distribution of different cell life cycles was determined with flow cytometry. Cell reproductive activities were detected with MTT method, and cell apoptesis was assessed with TUNEL. Results FasL protein was pigmentized at the position of 40 000 by Western blot, and the expression level of FasL was significantly higher in HR-8348F cells than those in HR-8348B, HR-8348L and HR-8348As cells(F=361.149, P<0.01) in normoxia. At 12 h after hypoxia, the FasL level was also significantly higher in HR-8348F cells than those in other groups (F=278.766, P<0.01), but was not markedly different as compared to themselves in normoxia (t=1.762, P>0.05). The proliferation index was significantly higher in HR-8348F (60.43±3.72) than those in HR-8348B (40.01±3.30), HR-8348L (41.30±4.06) and HR-8348As cells (35.87±4.39), respectively (F=39.477, P<0.01). However, both inhibition rate of proliferation and apoptotic index were remarkably lower in HR-8348F (17.30±1.98 and 13.10±1.04) than those in HR-834B (33.70±4.33 and 21.60±1.31), HR-8348L (34.20±3.92 and 20.10±1.15), and HR-8348As (38.00±4.55 and 23.90±1.23), respectively (F=28.811 and 76.462, respectively, P<0.01). Conclusion The expression enhancement of intracellular FasL in rectal carcinoma in hypoxia can lead to accelerated proliferation and reduced apeptosis of cells, which will promote tumor cells to adapt microenvironmental hypoxia.