目的通过检测食蟹猴皮下注射重组人干扰素α-2b后血清总抗体及中和抗体,研究其免疫原性。方法 16只健康食蟹猴,雌雄各半,雌性未孕,年龄3～4岁,给药开始时体重范围为:2.1～3.5 kg,按性别、体重随机分为4组,分别为对照组(0μg·kg-1),5,15和50μg·kg-1组,每组动物雌雄各半,每天1次颈背部皮下注射给药,共给药67天,恢复期14 d,于给药前,给药后第7天、14天、21天、28天、42天、56天、66天及恢复期末分别采集血样,并对给药前,给药后第7天、14天、21天、28天、42天和恢复期末的血样进行总抗体和中和抗体检测,抗体检测方法分别为:总抗体检测选择用ELISA的间接法;中和抗体检测采用细胞病变抑制法。结果总抗体:自给药14 d开始,低、中、高剂量组各有部分动物血清中产生猴抗重组人干扰素α-2b抗体;至给药42 d,给药组12只动物除低剂量组1只动物和高剂量组1只动物未产生猴抗重组人干扰素α-2b抗体外,其他10只动物均产生猴抗重组人干扰素α-2b抗体。经检测,产生的抗体为中和抗体,恢复14 d,抗体仍存在,但滴度有所降低。结论本试验条件下,低、中、高剂量组食蟹猴皮下注射重组人干扰素α-2b后,血清中可产生猴抗重组人干扰素α-2b抗体,产生的抗体为中和抗体。恢复14 d,抗体仍存在,但滴度有所降低。

良好农业规范(Good agricultural practice,GAP)是建立在现有标准和行业管理法律法规基础上,按照国际标准的基准框架构建起来的一个完整体系.其运用危害分析与关键控制点(HACCP)原理[1],对种植、养殖过程中食品安全、环境保护、员工安全和福利、动物福利4个方面的危害进行分析,并根据风险程度分为3级控制点进行有效控制,从而实现对农业生产源头全面、有效的控制.GAP是现代农业标准、法律和行业新技术的提炼,代表了行业发展的方向.

Objective:To study the toxic effects of zidovudin on rat hepatocytes in primary culture.Methods:The hepatocytes were isolated from Wistar rats with a two-step perfusion technique and the hepatocellular viability were tested with trypan blue exclusion.The hepatocytes which had more than 85% viability were used in the experiment.The experiment was divided into four groups as follows:the medication group,the positive control group,the negative control group,and the blank control group.The hepatocyte suspension(3×105 cells/ml) were inoculated into culture plates:200 μl was inoculated into each well of 96 well culture plates,1ml was inoculated into each well of 24 well culture plates,2.5 ml was inoculated into each well of 6 well culture plates.The hepatocytes were cultured in the incubator with 5% carbon dioxide at 37℃ for 4 hours,then the supernate was discarded.The hepatocytes were exposed to zidovudine 10,5,3.3,2,0.4,and 0.08 mmol/L in the medication group,carbon tetrachloride 10,2,and 0.5 mmol/L in the positive control group,1% DMSO-containing DMEM in the negative control group,and DMEM in the blank control group.The activity of hepatocytes in the 96 well plates were measured with MTT experiment 6,12,and 24 hours after exposure.The amounts of AST,ALT,and LDH release from hepatocytes in the 24 well plates were measured 6 and 12 hours after exposure.The hepatocytes in 6 well plates were stained with HE,and morphological changes were observed 12 hours after exposure.Results:The optical density(OD)to zidovudine(3.3 mmol/L) and carbon tetrachloride(2 mmol/L) 6,12,and 24 hours after exposure was(1.20±0.17) versus(0.99±0.08),(0.89±0.09) versus(1.20±0.13),and(1.01±0.09) versus(0.88±0.13),respectively.The OD in the negative control group was(1.34±0.08),(1.11±0.10),and(1.03±0.11),respectively,at the same time points.The hepatocyte activity in the zidovudine and carbon tetrachloride groups,compared with the negative control group,markedly decreased.The difference was statistically significant(all P<0.05).The effect of zidovudine 10 mmol/L on hepatocyte activity was lower than that of carbon tetrachloride 10 mmol/L.Six and twelve hours after exposure to zidovudine 3.3 mmol/L,the amounts of AST release were(18.53±2.02) KarU and(26.86±2.61) KarU,the amounts of ALT release were(15.16±2.18) KarU and(27.48±2.27) KarU,and the amounts of LDH release were(1 681.00±193.98) U/L and(2 708.55±78.73) U/L,respectively.In the negative control group,the amounts of AST release were(15.91±1.62) KarU and(37.71±2.54) KarU,the amounts of ALT release(19.66±0.74) KarU and(23.42±1.46) KarU,and the amounts of LDH release were(2 036.39±134.56) U/L and(2 870.21±87.73) U/L,respectively,at the same time points.Compared with the negative control group,zidovudine could significantly effect the amounts of AST,ALT,and LDH release.The difference were statistically significant(all P<0.05).The membrance rupture and karyopknosis of hepatocytes were occurred after exposure to zidovudine ≥3.3 mmol/L.And after exposure to carbon tetrachloride 10 mmol/L,the hepatocytes become smaller,and the membrance rupture and karyopyknosis of hepatocytes occurred.Conclusion:Zidovudine ≥3.3 mmol/L has hepatocellular toxicity,but its toxicity is less than carbon tetrachloride.

集约化生产条件下,肉鸽的生产性能不仅受日粮营养水平[1]、气候、生殖生理等综合因素作用的影响,而且对场地生产设施的摆放等因素也有特别要求.

Objective To observe the toxicity of cyclophosphamide(CP) on primary cultured hepatocytes and the protective effects of Oleanolic acid(OA). Methods Rat hepatocytes were isolated for primary culture and intracellalar ALT、AST and LDH leakages were detected in supernatant and MTT assay was used to evaluate the toxicity of CP and the protective potent of OA. Results CP increased the leakages of ALT,AST and LDH from hepatocytes with decreased the MTT value of hepatocytes.These changes were ameliorated partly by OA. Conclusion OA can protect hepatocytes from CP induced damage.

Objective: To compare the differences of collagenase and trypsin on isolated rat hepatocytes including quantity,viability, primarily culture and MTT experiment. Method: The hepatocytes was isolated and harvested by collagenase and trypsin from Wistar rats. The viability of hepatocytes was tested by trypan blue exclusion and the quantity was calculated at the same time. The morphological change of primarily cultured hepatocytes was observed. The proliferative hepatocytes was tested by MTT experiment. Result: The isolated hepatocytes quantity by collagenase was 2.4 times higher than trypsin, but the isolated hepatocytes viability by trypsin was 2. 8 times greater than collagenase. The isolated hepatocytes by collagenase grew slowly,but the isolated hepatocytes by trypsin grew fast. MTT experiment showed the activity of isolated hepatocytes by trypsin was greater than collagenase obviously. Conclusion: The quantity and purity of hepatocytes by collagenase was superior than trypsin, but the viability of hepatocytes by trypsin was better than collagenase.

Objective: To establish the acute alcoholic liver injury models (in vivo and in vitro) for further study on protective effects of injured liver by drugs. Method: The half lethal dose of ethanol was measured, the activity change of serum alanine transami-nase (ALT) and aspartate transaminase (AST) by alcohols with different doses and administrations were observed. The primary cultured hepatocytes were injured by ethanol,and the cultural supematants lactic acid dehydrogenase(LDH) release was inspected at different time spots and doses. Result: The half lethal dose for ethanol oral administration in mice was 9. 98 g·kg-1. Orally and intraper-itoneally administered ethanol seemed to increased serum ALT and AST when the doses were higher than 6.4 g·kg-1 and 4.0 g·kg-1 respectively. The LDH in cultural supematants was increased after 50-400 mmol·L-1 ethanol exposure to rat hepatocytes. Conclusion: The optimal injured dose of ethanol in vivo and in vitro were 7.2 g·kg-1 and 50-100 mmol·L-1 respectively.