Clinical and molecular analysis of hereditary non-polyposis colorectal cancer in Chinese colorectal cancer patients

AIM:To analyze the frequency of hereditary non-polyposis colorectal cancer(HNPCC)in Chinese colorectal cancer(CRC)patients,and to discuss the value of microsatellite instability(MSI)and/or immunohistochemistry(IHC)for MSH2/MLH1 protein analysis as pre-screening tests in China.METHODS:The Amsterdam criteriaⅠandⅡ(clinical diagnosis)and/or germline hMLH1/hMSH2 mutations(genetic diagnosis)were used to classify HNPCC families.Genetic tests,including microsatellite instability,immunohistochemistry for MSH2/MLH1 proteins and hMSH2/hMLH1 genes,were performed in each proband.RESULTS:From July 2000 to June 2004,1988 patients with colorectal cancer were analysed and 114 CRC patients(5.7%)from 48 families were categorized as having HNPCC,including 76 from 26 families diagnosed clinically and 38 from the other 22 families diagnosed genetically.The sensitivity and specificity of high MSI and IHC for predicting mutations were 100% and 54%,and 79% and 77%,respectively.CONCLUSION:The frequency of HNPCC is approximately 10% among all Chinese CRC cases.The MSI and IHC detections for hMSH2/hMLH1 proteins are reliable pre-screening tests for hMLH1/hMSH2 germline mutations in families suspected of having HNPCC.     

Augmented regeneration of partial liver allograft induced by nuclear factor-κB decoy oligodeoxynucleotides-modified dendritic cells

AIM: To investigate the effect of NF-κB decoy oligodeoxynuleotides (ODNs) - modified dendritic cells (DCs)on regeneration of partial liver allograft.METHODS: Bone marrow (BM)- derived DCs from SD rats were propagated in the presence of GM-CSF or GM-CSF+IL4 to obtain immature DCs or mature DCs, respectively. GMCSF-propagated DCs were treated with double-strand NF-κB decoy ODNs containing two NF-κB binding sites or scrambled ODNs. Allogeneic (SD rat to LEW rat) 50% partial liver transplantation was performed. Normal saline (group A),GM-CSF -propagated DCs (group B), GM-CSF+IL-4 -propagated DCs (group C), and GM-CSF+NF-κB decoy ODNs (group D) or scrambled ODNs -propagated DCs (group E)were injected intravenously into recipient LEW rats 7 days prior to liver transplantation and immediately after transplantation. DNA synthesis (BrdU labeling) and apoptosis of hepatocytes were detected with immunostaining and TUNEL staining postoperative 24 h, 48 h, 72 h and 84 h,respectively. Liver graft-resident NK cell activity, hepatic IFN-y mRNA expression and recipient serum IFN-γ level at the time of the maximal liver allograft regeneration were measured with 51Cr release assay, semiquantitative RT-PCR and ELISA, respectively.RESULTS: Regeneration of liver allograft was markedly promoted by NF-κB decoy ODNs-modified immature DCs but was significantly suppressed by mature DCs, the DNA synthesis of hepatocytes peaked at postoperative 72 h in group A, group B and group E rats, whereas the DNA synthesis of hepatocytes peaked at postoperative 84 h in group C rats and 48 h in group D rats, respectively. The maximal BrdU labeling index of hepatocytes in group D rats was significantly higher than that in the other groups rats.NF-κB decoy ODNs-modified immature DCs markedly suppressed but mature DCs markedly promoted apoptosis of hepatocytes, liver-resident NK cell activity, hepatic IFN-γmRNA expression and recipient serum IFN-γ production. At the time of the maximal regeneration of liver allograft, the minimal apoptosis of hepatocytes, the minimal activity of liver-resident NK cells, the minimal hepatic IFN-γ mRNA expression and serum IFN-γ production were detected in group D rats. The apoptotic index of hepatocytes, the activity of liver- resident NK cells, the hepatic IFN-γ mRNA expression level and the serum IFN-γlevel in group D rats were significantly lower than that in the other groups rats at the time of the maximal regeneration of liver allograft.CONCLUSION: The data suggest that the augmented regeneration of partial liver allograft induced by NF-κB decoy ODNs-modified DCs may be attributable to the reduced apoptotic hepatocytes, the suppressed activity of liverresident NK cells and the reduced IFN-γ production.     

Prolongation of liver allograft survival by dendritic cells modified with NF-κB decoy oligodeoxynucleotides

AIM: To induce the tolerance of rat liver allograft by dendritic cells (DCs) modified with NF-κB decoy oligodeoxynucleotides (ODNs).METHODS: Bone marrow (BM)-derived DCs from SD rats were propagated in the presence of GM-CSF or GM-CSF+IL-4 to obtain immature DCs or mature DCs. GM-CSF+IL-4-propagated DCs were treated with double-strand NF-κB decoy ODNs containing two NF-κB binding sites or scrambled ODNs to ascertain whether NF-κB decoy ODNs might prevent DC maturation. GM-CSF-propagated DCs, GM-CSF+NF-κB decoy ODNs or scrambled ODNs-propagated DCs were treated with LPS for 18 h to determine whether NF-κB decoy ODNs could prevent LPS-induced IL-12 production in DCs. NF-κB binding activities, costimulatory molecule (CD40, CD80, CD86) surface expression, IL-12 protein expression and allostimulatory capacity of DCs were measured with electrophoretic mobility shift assay (ENSA),flow cytometry, Western blotting, and mixed lymphocyte reaction (MLR), respectively. GM-CSF-propagated DCs, GM-CSF+IL-4 -propagated DCs, and GM-CSF+NF-κB decoy ODNs or scrambled ODNs-propagated DCs were injected intravenously into recipient LEW rats 7 d prior to liver transplantation and immediately after liver transplantation.Histological grading of liver graft rejection was determined 7 d after liver transplantation. Expression of IL-2, IL-4 and IFN-γ, mRNA in liver graft and in recipient spleen was analyzed by semiquantitative RT-PCR. Apoptosis of liver allograft-infiltrating cells was measured with TUNEL staining.RESULTS: GM-CSF-propagated DCs, GM-CSF+NF-κB decoy ODNs-propagated DCs and GM-CSF+ scrambled ODNs-propagated DCs exhibited features of immature DCs, with similar low level of costimulatory molecule(CD40, CD80,CD86) surface expression, absence of NF-κB activation,and few allocostimulatory activities. GM-CSF+IL-4-propagated DCs displayed features of mature DCs, with high levels of costimulatory molecule (CD40, CD80, CD86) surface expression, marked NF-κB activation, and significant allocostimulatory activity. NF-κB decoy ODNs completely abrogated IL-4-induced DC maturation and allocostimulatory activity as well as LPS-induced NF-κB activation and IL-12 protein expression in DCs. GM-CSF+NF-κB decoy ODNs-propagated DCs promoted apoptosis of liver allograft-infiltrating cells within portal areas, and significantly decreased the expression of IL-2 and IFN-γ mRNA but markedly elevated IL-4 mRNA expression both in liverallograft and in recipient spleen, and consequently suppressed liver allograft rejection, and promoted liverallograft survival.CONCLUSION: NF-κB decoy ODNs-rnodified DCs canprolong liver allograft survival by promoting apoptosis of graft-infiltrating cells within portal areas as well as down-regulating IL-2 and IFN-γ mRNA and up-regulating IL-4 rnRNA expression both in liver graft and in recipient spleen.     

CHA2DS2-VASc评分对急性心肌梗死患者院内结局事件的预测价值——China PEACE回顾性急性心肌梗死研究

目的评估CHA₂DS₂-VASc评分对急性心肌梗死(AMI)患者院内结局事件的预测价值。方法回顾性分析冠心病医疗结果评价和临床转化研究(China PEACE)回顾性急性心肌梗死研究中23728例AMI患者的病历信息,按CHA₂DS₂-VASc评分分为低(0~3分)、中(4~6分)、高(7~9分)分值组。院内结局包括主要不良心血管事件、死亡、死亡或放弃治疗、再发心肌梗死、缺血性卒中等。采用多因素Cox回归分析CHA₂DS₂-VASc评分对AMI患者院内结局的影响。通过受试者工作特征(ROC)曲线,评估CHA₂DS₂-VASc评分对AMI患者院内死亡与死亡或放弃治疗的预测价值。结果入组患者年龄66(56,75岁)岁,女性占30.7%。CHA₂DS₂-VASc评分高分值组患者院内结局事件发生率更高,基础疾病更多(P值均<0.001);多因素logistic回归中,院内病死率(OR=6.13,95%CI 4.77~7.87,P<0.001)、院内死亡或放弃治疗率(OR=6.43,95%CI 5.16~8.00,P<0.001)、主要心血管事件发生率(OR=4.94,95%CI 4.06~6.01,P<0.001)明显高于其他两组。ROC曲线分析显示,无论院内病死率,还是死亡或放弃治疗率,CHA₂DS₂-VASc评分与简化版全球急性冠状动脉事件登记(global registry of acute coronary events,mini-GRACE)评分相比差异无统计学意义(ROC曲线下面积:0.699与0.696,P=0.752;0.708与0.713,P=0.489)。结论CHA₂DS₂-VASc评分是一种有效预测AMI患者院内风险的评估工具,该评分操作简单,预测价值与mini-GRACE评分相当。     

DNA methylation profiling of ovarian carcinomas and their in vitro models identifies HOXA9, HOXB5, SCGB3A1, and CRABP1 as novel targets

Background  

The epigenetics of ovarian carcinogenesis remains poorly described. We have in the present study investigated the promoter methylation status of 13 genes in primary ovarian carcinomas (n = 52) and their in vitro models (n = 4; ES-2, OV-90, OVCAR-3, and SKOV-3) by methylation-specific polymerase chain reaction (MSP). Direct bisulphite sequencing analysis was used to confirm the methylation status of individual genes. The MSP results were compared with clinico- pathological features.     

无髓牙冠内漂白方法的对比研究

目的探讨无髓牙内漂白技术中2种细节操作方法对漂白效果影响。方法选择口腔中前牙和双尖牙区至少2颗变色无髓牙的患者40例,对2颗牙分别采用方法Ⅰ(标准根管治疗后,用光固化玻璃离子封闭根管口,用常温下浸有30%过氧化氢的棉球封闭在髓腔内,3d复诊1次,共封药3次后进行充填治疗)和方法Ⅱ(标准根管预备后,根充前用加热到36℃的30%过氧化氢反复冲洗髓腔,然后做常规根充和充填治疗)治疗。2组均采用同种复合树脂严密充填,1 a后用相同方法观察美观效果和强度效果。结果方法Ⅱ美观效果与强度效果均明显优于方法Ⅰ(P<0.05)。结论漂白变色的无髓牙最好方法是即刻高效的热漂白方法。     

水解猪全血蛋白提取氨基酸混合物的营养评价

猪全血经酸水解及离子交换树脂分离提取,适当补充L-色氨酸,L-蛋氨酸及L-异亮氨酸的混合氨基酸粉(简称配方Ⅰ)经蛋白质营养效价的研究结果表明,混合氨基酸配方Ⅰ,必需氨基酸含量为49.33％,其组成模式接近酪蛋白或全鸡蛋蛋白质,第一限制氨基酸为含硫氨基酸,化学分(CS)为63.77,必需氨基酸指数(EAAI)为92.78,均高于酪蛋白,大鼠生长试验证明,体重增长,蛋白质效能比(PER)和净蛋白比(NPR)与酪蛋白比较均无显著差别(P>0.05),但优于猪血粉(P<0.001)。本品可做为营养要素膳的良好氮源,又可以用于食品添加,为猪血的综合利用提供了一个重要途径。     

应用K-ras基因突变诊断胰腺癌的探讨

目的 采用连续富集酶切一限制性片段长度多态性／聚合酶链反应（CED-RnP／PCR）探索K-ras基因在胰腺癌患者血浆中的表达。方法 以血浆代替以往的标本并采用CED-RFLP／PCR技术检测胰腺癌、非胰腺癌患者及健康对照者血浆K-ras基因突变情况。结果 胰腺癌患者血浆K-Fas基因的突变阳性率为73％，无假阳性率，高于胰液、十二指肠液的检测率，低于细针穿刺的检测率；胰腺良性病变患者及健康对照者血浆中未检测到K-ras基因的突变；胰腺癌组K-ras基因突变与非胰腺癌组及健康对照组相比具有显著性差别。结论 CED-RFLP／PCR法检测血浆K-ras基因突变技术具有简单、易行高效的特点，克服以往技术的弊端，对胰腺癌的诊断及鉴别有一定的应用价值。     

康胃方对新生大鼠HP感染的根除及胃黏膜保护作用

目的 :HP悉尼菌株灌胃复制感染动物模型 ,观察康胃方对HP的根除及胃黏膜细胞保护作用。方法 :SD新生大鼠随机分为正常对照组、病理模型组、三联治疗组及康胃方组 ,除正常对照组外 ,每日经口接种 10 9CFU mlHP菌液 0 1ml,共 3次。 8周后进行UBT ,随即接受相应的治疗 ,共 4周。疗后第 4周 ,测定胃黏膜RO、NO、iNOS、MnSOD及NPSH的含量或活性。结果 :HP感染动物 ,UBT与胃黏膜所含RO、NO、iNOS和MnSOD明显升高 ,NPSH显著下降 (P <0 0 1)。康胃方治疗后 ,UBT降低 ,其他指标明显改善 (P <0 0 1)。结论 :康胃方对新生大鼠感染HP有明显的根除作用 ,并对胃黏膜有一定的细胞保护作用。