Objective :To investigate and compare the effects of different testing instruments and reagent combinations on PT and APTT coagulation detection ,reflect the differences in different detection systems ,to provide reference basis for data comparison for laboratories .Methods :Based on the different detection systems in varied manufacturers ,instru‐ments and reagents or different types sub‐group in the same manufacturers and instruments ,non‐parametric test and ANOVA rank method for statistical analysis ,by organized the data of PT and APTT in CGL sample from different la‐boratories issued by CAP in 2013 .To investigate and compare the effects of different testing instruments and reagent combinations .Results:There are different results of PT and APTT in varied detection systems .No significant difference exists in varied instrument manufacturers or reagents ,but there are significant difference in the results of the same manufacturer with different type reagents .On the contrary ,the results showed no significant difference in the same manufacturer with different type instrument .Conclusion:Manufacturer with the same instruments and reagents corre‐sponding detection results have good comparison .Construction of the data system available for reference in blood coag‐ulation detection is very important .

目的 评价氯胺酮的线粒体毒性,从而探讨氯胺酮可能的毒性机制.方法 HepG2细胞培养基中分别加入齐多夫定10～2000 μmol·L-1培养7d,或加入氯胺酮50～3000 μmol·L-1培养24 h,CCK8细胞计数法测定细胞存活率.HepG2细胞培养基中分别加入齐多夫定100 μmol· L-1培养7d后,或加入氯胺酮100和1200 μmol·L-1培养24 h,化学发光法检测胞内ATP合成水平,荧光探针法检测胞内钙离子浓度、活性氧及线粒体膜电位(△Ψm)的变化,同时设齐多夫定1000 μmol· L-1用于实时荧光定量PCR检测线粒体DNA(mtDNA)表达水平的变化.结果 与正常对照组相比,齐多夫定10～ 2000 μmol· L-1可以抑制细胞存活,IC50为100 μmol·L-1;氯胺酮1000～ 3000 μmol· L-1抑制细胞存活,IC50为1200 μmol·L-1.与正常对照组相比,齐多夫定100 μmol·L-1组线粒体ATP合成水平显著下降(P＜0.05),胞内钙离子浓度明显升高(P＜0.05),活性氧水平显著升高(P＜0.05),△Ψm显著下降(P＜0.05),而1000 μmol· L-1可以明显干扰mtDNA表达水平(P＜0.05).氯胺酮100 μmol· L-1组ATP合成水平明显下降(P＜0.05),其他指标均无显著性差异;氯胺酮1200 μmol·L-1组ATP合成水平显著下降(P＜0.01),活性氧水平和胞内钙离子浓度显著升高(P＜0.01),△Ψm显著下降(P＜0.05),mtDNA水平无明显改变.结论 氯胺酮可以通过干扰线粒体代谢功能而诱导线粒体损伤.

OBJECTIVE To explore the changes and role of plasma microRNA(miR-122)in non-clini-cal evaluation of drug-induced liver injury.METHODS Male SD rats were ig given acetaminophen(APAP)0(control),625 and 1250mg·kg-1.At 1.5,3,6,12,24,36 and 96h after administration,blood samples were collected.miR-103,as a suitable endogenous reference gene for plasma microRNAs quanti-fication in rats with acetaminophen-induced hepatotoxicity,was employed to detect the expression of miR-122 in plasma samples with real-time quantitative reverse transcription PCR(RT-qPCR).The blood levels of glu-tamic pyruric transminase(GPT)and glutamic oxaloacetic trnasminase(GOT)were detected and patho-logical changes in liver tissue were observed.RESULTS Histopathological examination showed that,compared with normal control group,there was no significant difference at 1.5 and 3h after rats were ig given APAP 625 and 1250mg·kg-1.Vacuolated hepatocytes and congestion in sinusoids were observed in zoneⅢat 6 and 12h after administration,while at 24h centrilobular necrosis was prominent in APAP 625 and 1250mg·kg-1 groups.Compared with normal control group,GPT and GOT in serum significantly in-creased at 12 and 24h in APAP 625 and 1250mg·kg-1 groups,and GPT activity more than doubled in APAP 1250mg·kg-1 group.Compared with normal control group,plasma miR-122 increased by 5.2-fold at 6h after administration in APAP 625mg·kg-1 and 3.6-fold at 1.5h after administration in APAP 1250 mg·kg-1.The level of plasma miR-122 continued to elevate but returned to normal at 36h in APAP 625 mg·kg-1 group and at 96h in APAP 1250mg·kg-1 group,respectively.The course of plasma miR-122 levels displayed similar kinetics to GPT and GOT.CONCLUSION miR-122 Expression in plasma may be-come an ideal hematologic molecular marker for early detection of drug-induced liver injury in pre-clinical and clinical evaluation.

Objective To investigate the nonclinical research facilities certified by SFDA,so as to know the developing and operating situation of GLP facilities.Methods To do the questionnaire survey on the Internet,this survey collected the detail information about the scale of the instrument of GLP facilities in China,staff and training situation,etc.and use SPSS to analyse the information.Results Thirty-eight SFDA-certified facilities provided valid information.The result of the survey shows that animal room's area of 90% facilities are above 500 m2;the number of staffs who have the bachelor degree is above 1600,the 30% staffs in half of facilities have master degree;30 facilities established animal welfare system;28 facilities completed the 3Q confirmed program of important monitoring instruments;7 facilities passed the CNAS ISO17025 of laboratory accreditation.Conclusion The operating and management standard of the nonclinical facilities are on the rapid developing stage,and that is close to the international standard on the side of personnel structure,animal management,instrument,software application and etc.

Objective To investigate the diurnal variations of hematological and biochemical parameters in cynomolgus monkeys.Methods The blood samples were collected in 12 cynomolgus monkeys(6 of each sex) at 9:30(am) ～ 10:30(am),12:30(pm) ～ 13:30(pm),15:30(pm) ～ 16:30(pm) and 19:30(pm) ～ 20:30(pm).The hematological and biochemical parameters were determined with automated hematological and biochemical analyzers respectively.The plasma cortisol levels were determined by high-performance liquid chromatography(HPLC).Results White blood count(WBC) and blood urea nitrogen(BUN) increased significantly in the afternoon in male and female monkeys.Hemoglobin(HGB) and hematocrit(HCT) markedly reduced in the afternoon in female monkeys.The activities of asparate aminotransferase(AST) and alanine aminotransferase(ALT) significantly increase in the afternnon in female monkeys.The serum glucose and plasma cortisol reduced significantly in the afternoon in male monkeys.Conclusions Parts of the hematological and biochemical parameters show obvious diurnal variation in male and female cynomolgus monkeys.

目的 探讨小鼠经口灌胃纳米二氧化钛后的急性毒性反应.方法 以5 g/kg剂量分别用纳米二氧化钛和微米二氧化钛单次经口灌胃染毒小鼠,观察14 d后处死,比较纳米二氧化钛和微米二氧化钛对小鼠体重、摄食量、血清生化指标及脏器系数的影响.结果 纳米二氧化钛对小鼠体重、脏器系数没有明显影响;实验期间,小鼠的摄食量逐渐增加;纳米二氧化钛染毒雌性组天冬氨酸转氨酶(AST)活力明显低于微米二氧化钛组,差异有统计学意义(P＜0.01),且肌酐(Cr)、尿素氮(BUN)与微米二氧化钛组、对照组相比,差异亦有统计学意义(P＜0.05).微米二氧化钛雌性组肌酸激酶(CK)活力明显高于对照组;纳米二氧化钛雄性组CK、乳酸脱氢酶(LDH)、α-羟丁酸脱氢酶(α-HBDH)活力明显高于微米二氧化钛组,微米二氧化钛雄性组LDH、α-HBDH活力明显低于对照组,纳米二氧化钛雄性组CK活力明显高于对照组,差异均有统计学意义(P＜0.05).结论 纳米二氧化钛染毒未见明显的急性毒性反应,但对血液生化指标可能有一定的影响,且与微米二氧化钛的影响不同.

Objective:To investigate the effects of vertilmicin and gentamicin on gene expression of human renal tubular epithelial cell lines(HK-2).Methods: Gene expression profiles were identified by DNA microarrays in HK-2 cells exposed to vertilmicin and gentamicin(3.2mg·mL-1) for 48h,respectively.Results: Vertilmicin significantly upregulated 87 genes and downregulated 10 genes of HK-2 cells.Gentamicin significantly upregulated 62 genes and downregulated 18 genes of HK-2 cells.The expressions of the various genes modulated cell growth and proliferation,apoptosis,cell cycle,stress response,transporters,chaperones,transcription regulation and drug metabolism.Conclusion: The alteration of these specific genes that regulated cellular apoptosis,stress response and transportation may be associated with cytotoxicity of aminoglycosides.

目的探讨单光子发射计算机体层显像仪(SPECT)技术在药物非临床研究—实验动物肾功能检测中的应用。方法犬静脉滴注某一类新药抗生素的长期毒性试验中,以血清生化检测、尿检、病理组织学检测(包括超微结构)和SPECT方法检测动物肾脏功能受损情况,对检测的结果进行汇总和比较。结果以SPECT检测GFR,结果显示在给药6周后各给药组与对照组差异显著,而在停药4周后,中、高剂量组GFR仍较大幅度低于对照组。生化结果亦显示给药6周时高剂量组CCR较低,但低、中剂量组不明显,而在停药4周时,见低、高剂量组低于对照,中剂量组反而高于对照组。有1例低剂量组动物于给药6周时生化、尿液及光镜检查均未发现明显异常,但SPECT检查结果显示其GFR低于对照组,以电镜对该动物进行肾细胞超微结构检查,发现线粒体嵴断裂、基质变空,自生性溶酶体多,间质纤维增生等现象,支持SPECT检测的结果。结论肾脏的代偿功能很强,所以当肾小球滤过功能已经降低时,SCR水平仍可在正常范围,直至肾脏已广泛破坏,SCR的变化才显现出来,此时SCR对测定肾功能衰竭的进一步发展才比较准确。从各项检查数据看,生化方法因为要收集动物24h尿液,可能存在样品污染,因此检测灵敏度和稳定性不及SPECT;同时一些细胞功能出现早期变化时,病理组织检查往往不易发现,而SPECT检测GFR有准确、快速和灵敏的特点。

目的本实验采用BLM诱导的大鼠肺纤维化模型,研究LEF对博莱霉素诱导的大鼠肺纤维化模型的影响作用,并与其他治疗类风湿性关节炎的药物进行比较。方法首先采用BLM制备大鼠肺纤维化模型,1次/d,经口灌胃给予LEF和地塞米松(Dexamethasone,DEX),同时设CMC溶剂对照组,连续给药4周。分别于给药后1、2和4周解剖大鼠,采集出肺血液,检测MDA和NO2-/NO3-的含量;采集肺组织,制备切片,分别采用HE、Masson三重染色和天狼猩红染色方法检查大鼠肺组织出现肺炎和肺纤维化的病理变化程度,采用组织芯片技术结合免疫组织化学方法检查TGF-β1在肺组织中不同部位的表达情况。结果在给药后1、2和4周,与CMC组相比,LEF、DEX组大鼠出现肺血液中MDA和NO2-/NO3-的含量明显降低,LEF、DEX组大鼠出现肺炎和肺纤维化程度明显降低;与CMC组相比,LEF、DEX组大鼠肺组织中TGF-β1表达阳性程度明显降低。结论研究结果提示,国内生产的来氟米特对博莱霉素诱导的大鼠肺纤维化模型的早期炎症反应具有一定抑制作用。

肾脏是体内最重要的排泄器官,也是外源性毒物在体内的主要代谢器官之一.很多化合物(药物)进入体内可引起肾脏损伤.国内外学者利用体内或体外模型对肾毒性作用及其机制进行了研究[1].近年来,毒理基因组学(toxicogenomics)的快速发展对毒性评价的模式、毒理学的研究方法产生了巨大影响.在肾毒性标志基因的鉴别、化合物肾毒性的量效关系、毒作用机制研究及肾毒性预测等方面,毒理基因组学已经提供或正在提供有价值的信息.现就毒理基因组学在肾毒性研究中的应用进展作一综述.

基因工程腺病毒H101是采用基因重组技术,通过对人5型腺病毒E1区改建重组的溶瘤性病毒.