交通协警多环芳烃暴露与健康风险的初步研究

目的 了解交通协警值勤期间多环芳烃个体暴露量,初步探讨此职业暴露所致增量致癌风险的水平.方法 于2005年7月26日-8月2日采集了天津市交通协警值勤期间空气PM10个体暴露样本,运用气相色谱-质谱(GC-MS)联用仪定量测定了空气中13种多环芳烃的含量,并与马路边和校园环境中多环芳烃浓度作对比;采用终生增量致癌风险模型对其职业暴露所致增量致癌风险进行了初步计算;并对风险计算进行灵敏度分析.结果 交通协警工作环境空气中颗粒态苯并[a]芘及13种多环芳烃浓度之和的均值分别为26、867 ng/m3,显著高于马路边和校园内浓度;其职业多环芳烃呼吸暴露所致致癌风险增量介于10-6～10-3之间;灵敏度分析显示,苯并[a]芘致癌强度因子和多环芳烃浓度是对风险计算结果影响最大的两个因素.应提高这两个因素取值的准确性来提高风险评价的准确性.结论 交通协警职业多环芳烃暴露存在一定的增量致癌风险.     

绿麦隆和阿特拉津联合染毒对小鼠睾丸的影响

目的 研究绿麦隆和阿特拉津单体系及联合作用对小鼠睾丸形态及结构的影响.方法 将昆明种小鼠按灌胃农药和剂量随机分为20组[1个对照组,4个绿麦隆染毒组(321.5、1250、2500、5000mg/kg),3个阿特拉津染毒组(218.75、875、1750mg/kg),12个联合染毒组],每组10只.连续25 d经口灌胃,每天1次,每次1ml.并用光学显微镜、电子显微镜对小鼠睾丸病理学变化进行观察.结果 农药单体系及联合染毒体系各剂量组对小鼠睾丸均有不同程度的损伤.与对照组比较,光学显微镜观察所见为各剂量组出现不同程度生精上皮细胞排列疏松、紊乱,生精细胞脱落,层次减少,病变严重;电子显微镜观察为各剂量组出现不同程度生精细胞线粒体呈空泡样改变,核膜肿胀、弯曲,支持细胞功能低下随染毒剂量增加,上述病理学变化有加重趋势,联合染毒组病理变化比单体系染毒组更明显.结论 绿麦隆和阿特拉津对小鼠睾丸的毒性与染毒剂量相关,联合作用体系加重了对小鼠睾丸的毒性效应.     

半夏多倍体复合体基因组DNA甲基化状态的MSAP分析

目的分析半夏多倍体复合体DNA甲基化状态的变化(甲基化水平和甲基化差异模式)。方法应用甲基化敏感扩增多态性(methylation sensitive amplified polymorphism,MSAP)技术,采用34对引物进行选择性扩增。结果扩增共得到7708条带,其中5636条带为甲基化多态性带在半夏7倍、8倍、9倍、10倍体值株间DNA甲基化水平变化不大。总扩增位点甲基化水平在54%～58%,全甲基化率为24.1%～24.3%,高倍性植株甲基化程序相应比较高。除甲基化水平稍有变化外,半夏不同倍性株间的DNA甲基化模式也形式各异,检测到的带型分为两大类(和型)。其中,不同倍性间DNA甲基化状态保持不变的位点占52.5%,归为型;少部分检测位点(占47.5%,归为型)的DNA甲基化模式在不同倍性间存在显著差异。结论半夏多倍体复合体各倍性植株间存在大量的胞嘧啶甲基化变异且整体甲基化水平较高。     

大孔吸附树脂对莲子心中有效成分的分离纯化研究

目的研究大孔吸附树脂吸附分离莲子心中莲心碱、异莲心碱及甲基莲心碱的工艺条件及参数。方法考察了AKS-W、AB-8和H214树脂对莲心碱、异莲心碱及甲基莲心碱的静态及动态吸附性能,并采用HPLC定量分析莲心碱、异莲心碱及甲基莲心碱。结果AKS-W树脂对莲子心中有效成分有较好的吸附分离效果,提取物中莲心碱、异莲心碱及甲基莲心碱的质量分数分别提高到10.2%、6.7%和11.9%。结论大孔型非极性吸附树脂AKS-W对莲子心中有效成分是一种较为理想的分离介质。     

The Effect of calmodulin antagonist berbaminederivative- EBB on hepatoma in vi tro and in vivo

Objective To evaluate the anti- hepatoma effect of Calmodulin antagonist 0- 4- ethoxyl- butyl- Berbamine (EBB), one of the berbamine derivatives. Methods Monotetrazolium (MTT) method was used to analysize the effect of EBB on the proliferation and growth inhibition effect Of a hepatoma cell line in vitro.A mouse hepatoma model was induced by injection of hepatoma ells (H22) in the abdominal cavity.The effect of EBB on survival at different concentrations as well as in combination with 5- FU were investigated in vivo.Flow cytometry analysis, dot blot hybridization, western blot, immunochemistry, enzyme- linked lectin assay (ELISA), trifluoperazine (TFP) and electron microscopic observation were used to study the effect of EBB on cell cycle process, P53 mRNA and protein levels, calmodulin content and ultrastractural changes of hepatome cells. Results EBB exerts a very strong inhibitory effect on human hepatoma cell line 7402 and mouse hepatoma cell line H22 in vitro.The IC(50) value of EBB for the two cell lines are 3.312 μg/ml and 1.167 μg/ml, respectively.The sensitivity of H22 cells to 5- FU can be markedly enhanced: The IC(50) dosage of 5- Fu can be decreased from 0.75 μg/ml down to 0.15 μg/ml, when jointly administered with nontoxic dosages of EBB (IC10 ).In vivo, EBB can prolong the lifespan of mice with ascites H22 to more than three months.64% of mice survived, while all animals in the control group died by the 18th day.When EBB (5 mg·kg (- 1) ·d( - 1) ) is jointly used with 5- FU (25 mg·ml - 1 ·d[ - 1 ), 73% of mice with ascites H22 survived, much higher than 27% in the 5- FU treated group.EBB can enhance the anti- hepatoma ability of 5- Fu treatment.EBB mechanism against hepatoma: P53 expression in the EBB treated group is substantially higher than that in the control group.EBB increased the translation of P53.As a calmodulin antagonist, EBB decreases amount of the CaM in hepatoma cells and blocked the hepatoma cell proliferation cycle at the G 2M phase.Before the G 0/G 1 phase, a diploid peak and apoptic cells in the treated groups were observed. Conclusions The CaM antagonist, EBB, has a strong anti- hepatoma effect and enhances the effect of 5- FU, induces hepatoma cell to apotopsis, promotes the P53 protein expression and decreases the amount of CaM in the cytoplasm.All these results demonstrate that EBB is a new and potentially useful drug against hepatoma and should be researched further.     

壳聚糖的抑菌活性分析及其抑菌机理的研究

制备10个分子量的壳聚糖，对6种细菌作抑菌实验。抑菌实验结果表明分子量在14．9，297．6ku的壳聚糖有很强的抑菌活性，此活性有随pH的降低而增加的趋势，pH5．5时最低抑菌浓度为0．03％-0．06％，Ca^2＋、Mg^2＋可显著降低壳聚糖对革兰阴性菌的抑菌作用。壳聚糖的抑菌机理有可能影响细胞膜的通透性。对实验结果的方差分析表明，壳聚糖的不同分子量、pH值和金属离子等外界因素都是壳聚糖抑菌活性的极显著影响因素，但菌株本身才是壳聚糖抑菌活性大小的最关键因素。     

病毒序列促转基因在斑马鱼胚胎发育过程中的高效表达

目的探讨转基因在斑马鱼胚胎发育过程中高效表达的有效途径和手段。方法以增强处理的绿色荧光蛋白(GFP)作为报告基因,在表达质粒的两翼连接构建腺关联病毒(adeno-assoc iated virus,AAV)来源的病毒性反转末端重复序列(inverted term inal repeats,ITR s),以显微注射方式注入斑马鱼受精卵的单一细胞和双细胞时相。结果病毒序列中的ITR s可使转基因表达效率达到33.0%,远远高于未与病毒序列结合的质粒。结论源于病毒的ITR s可显著促进转基因在斑马鱼胚胎发育过程中的表达。     

The Biological Effect of Hepsin on the Proliferation and Invasion of PC-3 Prostate Cancer Cells

OBJECTIVE Recent studies have shown that hepsin, a type of transmembrane serine protease, is highly upregulated in prostate cancer, but, little is known about its role in progression and invasion of this cancer. We constructed a hepsin-expressing plasmid and transfected it into PC-3 cells to investigate the effect of the hepsin gene on the biological behavior of the PC-3 cells. METHODS Plasmid pHepsin-IRES2 was transfected into prostate cancer PC-3 cells using Fugene6, and the cells with stable hepsin expression were screened and selected with Zeocin (600 mg/L). The hepsin mRNA level was measured by real-time PCR and the growth curve of the PC-3-transfected cells assessed using MTT and BrdU assays. A Boyden chamber was used to examine the difference in invasion and metastases between transfected and non-transfected cells. RESULTS The hepsin mRNA level in pHepsin-IRES2 transfected -PC-3 cells was significantly higher than that found in the control PC -3 cells. While the growth curve of the hepsin gene transfected PC -3 cells showed that there was no significant effect on proliferation, the invasive ability of the pHepsin-IRES2 transfected PC-3 cells, as compared with control cells, was significantly increased (P<0.05). CONCLUSION The results suggest that even though hepsin has no effect on the proliferation of prostate cancer PC-3 cells, it does promote cellular invasion and metastasis.Therefore hepsin may have a role in the development of prostate cancer.     

两种CPD光敏剂对小鼠S180肉瘤的光动力杀伤效应

目的：比较新型光敏剂CPD4和CPD6对小鼠S180移植肉瘤的光动力学疗效。方法：给昆明小鼠接种S180肉瘤，建立移植肿瘤模型。将模型小鼠随机分成7组（n=8），分别为CPD4和CPD6中、低剂量组，血卟啉组，生理盐水对照组及未经任何治疗组。尾静脉注射给药，给药后20min以波长670nm激光垂直照射瘤区20min（血卟啉组照射60min）。治疗后30d将各组小鼠断颈处死，取瘤称质量并计算抑瘤率，做病理切片、常规染色，镜下观察形态学差异。结果：CPD4中、低剂量组的抑瘤率分别为54．5％，97．9％；CPD6低剂量组的抑瘤率为46．9％；血卟啉组的抑瘤率为55．8％．镜下观察各组切片形态学差异明显。CPD。低剂量组的疗效高于血卟啉组，亦高于同剂量CPD6组。CPD6低剂量组疗效与血卟啉组相近。结论：光动力学治疗对小鼠S180移植肉瘤有抑制作用，该研究可为生产治疗恶性肿瘤的新药提供科学的实验依据。