小干扰RNA干扰大鼠神经元Nogo受体mRNA表达的实验研究

目的观察大鼠Nogo受体(NgR)特异性小干扰RNA(small interfering RNA,siRNA)在原代神经元干扰其mRNA表达的效果。方法体外培养大鼠原代皮层和海马细胞,应用阳离子脂质体转染试剂转染4对化学合成的大鼠NgR特异性siRNA,72h后提取细胞总RNA,实时荧光定量RT-PCR检测NgR mRNA表达情况。结果4对siRNA均能够下调靶基因NgR的mRNA表达水平,siNgR199、siNgR562、siNgR772和siNgR964等干扰后,NgR mRNA的表达分别为对照siRNA干扰组的6.5%、62.4%、15.2%和6.86%,与对照siRNA干扰组比较有统计学意义(P<0.05)。结论化学合成的NgR特异性siRNA能够有效的干扰大鼠原代皮层和海马细胞内NgR基因mRNA的表达水平。     

胰腺癌中p16基因甲基化改变及其蛋白表达分析

目的:探讨胰腺癌与癌旁组织中p16基因启动子区异常甲基化的改变及其蛋白表达的特点,以及其与胰腺癌发生发展的关系。方法:分别采用免疫组化SP法及甲基化特异PCR(MSP)检测46例人胰腺癌和癌旁组织中p16基因表达及其甲基化的水平,并结合临床资料进行分析。结果:胰腺癌中p16蛋白表达率为41.3%(19/46),而癌旁组织表达率为95.7%(44/46),两者有差异显著性(P<0.01)。p16蛋白阳性的19例胰腺癌标本中均未检出基因甲基化;p16蛋白缺失的27例标本中有18例检出基因甲基化,甲基化率为39.1%。p16基因甲基化与蛋白缺失有明显关系(P<0.05)。p16基因表达及其启动子区甲基化的发生率与胰腺癌的大小,患者性别﹑年龄的差异无显著意义(P>0.05),但与组织分化程度﹑淋巴结转移﹑PTNM分期有显著意义(P<0.05)。结论:p16基因启动子的异常甲基化可影响p16蛋白的表达,它们与胰腺癌的发生发展有关;p16甲基化和蛋白表达可能成为胰腺癌诊断及预后的候选标志物之一。     

Postmenopausal Women With Colles'' Fracture Have Lower Values of Bone Mineral Density Than Controls as Measured by Quantitative Ultrasound and Densitometry

Measurement of ultrasonographic parameters provides information concerning not only bone density but also bone architecture. We investigated the usefulness of ultrasonographic parameters and bone mineral density (BMD) to evaluate the probability of Colles' fracture. Two-hundred eighty-nine postmenopausal women (62.3 +/- 8.7 yr) with (n = 76) and without (n = 213) Colles' fracture were studied. BMD of lumbar spine and proximal femur was evaluated in all women by dual-energy X-ray absorptiometry (DXA) and speed of sound (SOS), broadband ultrasound attenuation (BUA), and stiffness in the calcaneus were measured by a Sahara ultrasonometer (Hologic). Patients suffering from Colles' fracture had lower values of BMD adjusted by height at the lumbar spine, L2-L4 (0.797 g/cm2 vs 0.860 g/cm2), femoral neck (0.685 g/cm2 vs 0.712 g/cm2 ), SOS (1518 m/sg vs 1525 m/sg), and stiffness (74.6 vs 77.7) (p < 0.05). Nevertheless, BUA values were similar in both groups. After stepwise logistic regression analysis, the area found under receiver operating characteristic (ROC) curves was 0.60 for L2L4 and 0.63 for a formula combining L2L4 and height. Our data suggest that patients suffering from Colles' fracture have lower values of BMD by DXA, SOS, and stiffness. However, the ability of these techniques to discriminate is low because the values for the area under ROC curve are 0.60 for L2-L4 and 0.63 for a formula derived of the combination of L2-L4 and height.     

Quantitative observation by enzyme cytochemistry of human rectal adenocarcinoma cells killed by LAK cells

Activity of succinic dehydrogenase(SDH)and acid phosphatase(AcPase)of effector-target cells during the process of LAK cells killing HR8348 cells was estimated by enzyme cyto-chemistry technique.SHD positive granules and AePase gray level were assayed with MIAS-300image analyser.The results showed:(1)After cocultivation of effector and target cells for vari-ous times,the activity of AcPase of HR8348 cells was apparently higher than that of the controlgroup,and it increased following prolonged coincubation.SDH activity of target cells increasedmarkedly within 30 and 60 rain cocultivation,but became low after 90 min treatment.(2)Ac-Pase content within LAK cells at 60,90,120,180 and 240 rain cocultivation was significantlyhigher than that of control group(P<0.01).The phenomenon of high AcPase and SDH activitywithin effector-target cells indicates that the function of the two types of cells was in an activestate.At the early stage of effector-target combining,the increase of SDH with HR8348 cellsmay be related to defensive function of the target cells.Higher AcPase activity of target cells in-dicates the activation of lysosomal enzyme which serves as the material basis for autolysis of thecells.     

Invasive techniques for the diagnosis of respiratory infectious diseases

Conclusion Reviewing the history of diagnostic procedures of causative organisms of respiratory infections, invasive techniques such as the protected specimen catheter (PSB) and bronchoalveolar lavage (BAL) have become the preferred choices because they have many advantages. These methods cause the patient relatively little discomfort, and permit an early diagnosis since they can easily be performed at the bedside and the causative organism from the disease site is obtained in cultures. These procedures can be used not only in patients with community-acquired lung infections, but also in immunocompromised hosts, including those with blood diseases or following renal transplantation, in patients in intensive care units and in mechanically-ventilated patients so that the cause can be accurately determined and chemotherapy started quickly, resulting in better therapeutic efficacy. Although these invasive procedures are advantageous for the diagnosis of respiratory infections, they also present various problems which remain to be addressed including minimizing contamination and setting diagnostic threshold values. However, the importance of accurately determining the causative organism in respiratory infections should be recognized as the most important factor, and these methods have shown to date to provide the most accurate information to aid in the timely treatment of respiratory infections in a wide variety of patients.     

P53、P16、Bcl-2基因及产物在原发性肺癌中的表达及其临床病理意义

目的 :探讨原发性肺癌 Mtp5 3、p16、Bcl- 2的异常表达与肺癌发生、发展的关系 ,以及它们之间的调控关系。 方法 :用免疫组化技术检测 (L SAB) 114例原发性肺癌组织中 p5 3、p16、Bcl- 2的表达。并用 PCR技术对 6 2例 p16蛋白丢失的肺癌组织中 p16外显子 2 (p16 E2 )的缺失进行分析。 结果:p5 3蛋白异常表达与肺癌组织学类型、分化程度无关 (P >0 .0 5 ) ,但与淋巴结转移情况有关 (P >0 .0 5 )。 p16蛋白阳性表达率与肺癌的组织学类型无关 ,但是其表达水平与非小细胞肺癌 (NSCL C)的细胞分化程度和淋巴结转移密切相关 (P <0 .0 5 )。 NSCL C中 p16E2的缺失率为 45 .2 %,其缺失水平随淋巴结转移和组织学分级的升高而升高 (P <0 .0 5 )。 SCL C(小细胞肺癌 )和正常肺组织中无 p16 E2的缺失。 Bcl- 2在小细胞肺癌中阳性率 6 2 .2 %显著高于 Sq Ca(鳞癌 )的 2 5 %和 Ad Ca(腺癌 )的 9.1%(P <0 .0 5 ) ,与细胞分化程度和淋巴结转移情况无关。另外 ,p5 3与 p16蛋白之间存在调控关系。 结论 :(1) p16、p5 3、Bcl- 2的异常参与肺癌的发生、发展过程。 (2 ) p16基因及产物的异常表达与 NSCL C的组织学分级和淋巴结转移相关 ,可能参与 NSCL C的发生、发展和转移过程。 (3) Bcl- 2反映 SCL C的分之生物学行为和临床     

LightCycler实时监测PCR定量分析血清HBV DNA

目的 检验LightCycler实时监测PCR（real-time detection PCR,RTD-PCR)对血清中HBV DNA定量检测的灵敏性和可重复性，探讨HBV血清标志物与HBV DNA定量的关系。方法 HBV定量按深圳匹基公司乙肝PCR荧光检测试剂盒使用说明，对乙肝标志物已明确的773例血清中HBV DNA定量结果进行统计分析。结果 实时监测PCR对血清中HBV DNA定量检测的灵敏性高，可检测低至1000拷贝／ml血清；可重复性好，批间误差＜20％；各种标志物类型的血清HBV DNA含量分布情况表明，HBV血清标志物中HBeAg与HBV DNA含量有明显的关系，一般HBeAg阳性血清HBV DNA含量较高，但也有相当一部分例外。结论 LightCycler实时监测PCR对血清中HBV DNA定量检测灵敏性高可重复性好；仅根据HBV血清标志物往往不能确定乙肝患者HBV DNA复制水平的高低，HBV DNA定量检测具有十分重要的临床意义。