叶酸对铅中毒幼年大鼠中枢神经系统的保护作用

通过研究铅对大鼠中枢神经元的损伤以及应用叶酸后的影响，探讨了叶酸对铅中毒条件下中枢神经元的保护作用。将健康清洁级初断乳3周龄SD大鼠随机分为铅染毒组，叶酸给药组及生理盐水组。各组于处理后第2、3、4周取材。铅染毒组、叶酸给药组每组各30只，生理盐水组12只，共72只。测量血铅，分别进行海马和小脑组织的H—E染色及凋亡抑制基因Bcl—2、凋亡基因Bax的蛋白免疫电泳，并作光密度分析。结果，通过对各个时间段各组血铅测定，可得叶酸给药组血铅含量低于铅染毒组。铅染毒组Bcl-2蛋白表达量少于给药组，而Bax多于给药组，H—E染色结果铅染毒组神经细胞数量明显少于叶酸给药组。结果表明，叶酸可降低铅中毒大鼠的血铅浓度，并具有一定抗细胞损伤作用，对铅中毒大鼠的中枢神经元具有一定的保护作用。     

Metabolism of flupyrazofos in the isolated perfused rat liver

To investigate the hepatic metabolism of the new insecticide flupyrazofos [O,O-diethyl O-(1-phenyl-3-trifluoromethylpyrazol-5-yl) phosphorothioate], isolated rat liver was perfused with flupyrazofos under single-pass conditions. In outflow perfusate and bile, 1-phenyl-3-trifluoromethyl-5-hydroxyprazole (PTMHP), PTMHP-sulfate and PTMHP-glucuronide conjugates were identified as the metabolites of flupyrazofos. However, O,O-diethyl O-(1-phenyl-3-trifluoromethylpyrazol-5-yl) phosphate (flupyrazofos oxon) was not detected. A HPLC method with UV detection was used to investigate the hepatic disposition of flupyrazofos and its metabolite PTMHP. The concentrations of flupyrazofos, PTMHP and PTMHP conjugates in outflow perfusate reached steady-state levels within 20 min after commencing perfusion of 7.3 microM flupyrazofos. At steady state, the mean extraction ratio of flupyrazofos was 0.93 (+/- 0.01) and clearance was 26.1 (+/- 0.2) ml min-1 which nearly approached perfusate flow rate (28 ml min-1). PTMHP accounted for 55.7 (+/- 5.8)% of eliminated flupyrazofos and was recovered as unchanged PTMHP, PTMHP-sulfate and PTMHP-glucuronide in the bile as well as the outflow perfusate.     

骨骼肌纤维烟酰胺腺嘌呤二核苷酸四唑氧化还原酶组化染色的检测

本文应用烟酰胺腺嘌呤二核苷酸四唑氧化还原酶（NADH-TR）组化染色，将大鼠与家兔小腿胫前肌与腓肠肌的肌纤维型分成染色最深的快缩氧化FO型、染色较深的慢缩氧化SO型、染色浅的快缩氧化酵解FOG型及染色最浅的快缩酵解FG型纤维。这与定量分析结果相吻合。并对低温液氮保存骨骼肌标本进行观察，发现小块肌组织＋生理盐水和小块肌组织＋5％二甲基亚砜（DMSO）冷冻保护剂的玻璃化冻存一起浸入液氮制成切片，都未见冰晶形成；而在用5％二甲基亚砜玻璃化冻存大块肌组织切片见有少量冰晶出现；未使用5％二甲基亚砜玻璃化冻存大块肌组织切片见有大量冰晶出现。提示小块标本超快速浸入液氮内能减少冰晶形成，保证酶组化染色切片的质量。并进一步对烟酰胺腺嘌呤二核苷酸四唑氧化还原酶组化作用机理与孵育液的科学配制进行了探讨。     

The Effect of Lecithin Supplementation on the Biochemical Profile and Morphological Changes in the Liver of Rats Fed Different Animal Fats

Sixty-four Wistar rats were divided into 8 equal groups and kept for 36 days in individual boxes. Three of the groups were given full synthetic diets containing various animal fats: beef tallow, pork fat or fish oil. A control group was fed the diets without the fat. The other four groups were fed the same diets with lecithin supplementation. At the end of experiment, sections taken from the liver were stained with haematoxylin–eosin and Sudan III to indicate fat infiltration. Liver enzyme levels, total bilirubin, albumin and two products of lipd metabolism were measured in serum from all the animals. The addition of lecithin to the diets did not influence the level of enzymes in the serum except in rats fed the diet containing beef tallow. A relationship between the type of diet, lecithin supplementation and the total cholesterol concentration in serum was observed. The concentration of HDL-cholesterol was only influenced by lecithin supplementation and that of triglycerides by the type of fat in the diet. The addition of lecithin to the diet was associated with proliferation of Kupffer cells, and an increased number of binuclear cells. Fatty degeneration of hepatocytes was less pronounced in all groups following lecithin supplementation. Lecithin supplementation of the diet did not elicit any pathological lesions and may be considered as a hepatocyte protector. This favourable effect of lecithin was most marked in the group of rats fed the fish oil.     

Modulation of sodium channels by the oxadiazine insecticide indoxacarb and its N-decarbomethoxylated metabolite in rat dorsal root ganglion neurons

The effects of the oxadiazine insecticide indoxacarb and its N-decarbomethoxylated metabolite (DCJW) on tetrodotoxin-resistant (TTX-R) voltage-gated sodium channels in rat dorsal ganglion neurons were studied using the whole-cell patch clamp technique. Indoxacarb and DCJW suppressed the peak amplitude of action potentials, and DCJW exhibited a faster time course and higher potency than indoxacarb in the blocking effects. In voltage-clamp experiments, indoxacarb and DCJW suppressed TTX-R sodium currents in a time-dependent manner without a steady-state level of suppression. IC50 values for indoxacarb and DCJW on TTX-R sodium currents were estimated to be 10.7 and 0.8 microM after 25 min of bath application, respectively. DCJW was about 10 times more potent than indoxacarb in blocking TTX-R sodium currents. Although the suppressive effects of indoxacarb were partially reversible after washout with drug-free external solution, no recovery of sodium current was observed in DCJW treated neurons after prolonged washout. In current-voltage relationships, both indoxacarb and DCJW blocked the sodium currents to the same degree in the entire range of membrane potentials. The sodium conductance-voltage curve was not shifted along the voltage axis by indoxacarb and DCJW at 10 microM. In contrast, the steady-state inactivation curves were shifted in the hyperpolarizing direction by indoxacarb as well as by DCJW. Based on these results, it was concluded that indoxacarb and DCJW potently blocked the TTX-R sodium channel in rat DRG neurons with hyperpolarizing shifts of the steady-state inactivation curves, suggesting preferential association of the insecticides to the inactivated state of sodium channels. The small structural variation between indoxacarb and DCJW resulted in clear differences in potency for blocking sodium channels and reversibility after washout.     

Possible Anxiolytic Effects of Ivermectin in Rats

Ivermectin, a mixture of 22,23-dihydroavermectin B_1a (80%) and B_1b (20%), is produced by Streptomyces avermectilis, an actinomycete. It is a macrocyclic lactone disaccharide, a member of the avermectin family, and is used as an antiparasitic drug. Previous studies performed in our laboratory showed that doramectin, another avermectin drug, interferes with GABAergic-related behaviours, leading to anxiety and seizures. The objective of the present study was to examine the effects of ivermectin (0.5 and 1.0 mg/kg) on the central nervous system of rats, using behavioural models related to GABAergic neurotransmission. A known anxiolytic drug, diazepam, was used as a positive control. Open field and elevated plus-maze behaviours, as well as conflict behaviour to a conditioned response, were assessed. The effects of ivermectin and diazepam in reversing the anxiety induced by picrotoxin was studied. The protective effects of ivermectin on pentylenetetrazole- and picrotoxin-induced seizures were also investigated. In the open field, 1.0 mg/kg ivermectin decreased locomotion frequency at 15 and 60 min of observation, rearing behaviour showed a biphasic effect at 15 and 30 min and duration of immobility was increased in all sessions after 1.0 mg/kg ivermectin. These data suggest anxiolytic or sedative effects. Ivermectin and diazepam both had a tendency to cause an increase both in the number of entries into the open arms and on the time spent in the open arms of an elevated plus-maze. Picrotoxin on its own reduced the number of entries as well as the time spent in the open arms. Both diazepam and ivermectin reversed these effects of picrotoxin. In conflict behaviour analysis, ivermectin and diazepam gave the classic effect of an anxiolytic drug, reversing the conditioned response to shock. Ivermectin protected rats from the convulsant effects of pentylenetetrazole but not from those of picrotoxin. Thus, ivermectin had the pharmacological profile of an anxiolytic drug with GABAergic properties. The lack of effect on seizures induced by picrotoxin suggests that the action of ivermectin is different from that of the benzodiazepine drugs.     

Dermal pharmacokinetics of the insecticide furathiocarb in rats

The pharmacokinetics of furathiocarb were studied in vivo in male Sprague-Dawley rats following dermal treatment. HPLC and post-column derivatization were used for the analysis of furathiocarb and its metabolites (carbofuran, 3-hydroxycarbofuran and 3-ketocarbofuran). Carbofuran and 3-hydroxycarbofuran were detected in plasma and urine rather than furathiocarb. 3-Ketocarbofuran, another potential metabolite, was not observed in any sample. The concentration of carbofuran was higher than that of 3-hydroxycarbofuran in plasma, but the reverse was the case in urine. The corresponding area under the plasma concentration-time curve, Tmax, and Cmax values of carbofuran and 3-hydroxycarbofuran for 1500 mg kg-1 doses were 2.4-8.0 mg equiv hml-1, 12 h and 0.1-0.4 mg equiv ml-1, respectively. T1/2 was calculated only for 3-hydroxycarbofuran (28 h). Two metabolites were excreted in a dose-dependent manner without saturation.     

乳鼠成骨细胞的培养及鉴定

无菌采取5日龄SD大鼠的颅盖骨,用胰蛋白酶预消化后,漂洗,剪碎,再用Ⅱ型胶原酶消化获得成骨细胞。通过倒置相差显微镜观察,并用碱性磷酸酶和矿化结节染色鉴定,利用微量滴定（MTT）法测定一个培养周期的成骨细胞活性分布,探讨一种经济、高效的原代成骨细胞培养方法。倒置相差显微镜形态观察结果显示,未贴壁的细胞呈圆形,贴壁生长的细胞呈不规则梭形、三角形或多角形,单核,有1-3个核仁;碱性磷酸酶和钙化结节染色均呈阳性（阳性率≥98.06%）;MTT法结合形态学观察结果显示,成骨细胞培养至第10 d时活性最强。证实用改良的二次酶消化法能够在短时间内获得大量成骨细胞,且所培养的细胞具有典型的成骨细胞形态和功能。     

半胱亚磺酸脱羧酶(CSD)基因在大鼠睾丸组织中的表达及其序列分析

试验根据大鼠肝脏牛磺酸生物合成关键酶——半胱亚磺酸脱羧酶(CSD)基因已知核苷酸序列,设计了1对特异的寡核苷酸引物,对提取的睾丸组织总RNA进行RT-PCR扩增,扩增出468 bp的DNA片段,在国际上首次证明CSD基因在大鼠睾丸组织中存在表达,即牛磺酸可在大鼠睾丸组织中生物合成。测定了扩增的睾丸组织CSD基因核苷酸序列,并与已知肝脏CSD基因核苷酸序列及其对应的氨基酸序列作了同源性分析。结果表明:睾丸组织CSD基因核苷酸序列同已知肝脏CSD基因核苷酸序列的同源性为99.8%,其对应氨基酸序列的同源性为100%。     

苜蓿雄性不育系组织培养技术的研究

以苜蓿雄性不育系Ms-4幼嫩茎段、叶片、叶柄为外植体,诱导苜蓿雄性不育系再生植株.结果表明:叶柄是诱导愈伤组织的良好外植体,体细胞胚诱导率为90%,再生率49%.苜蓿雄性不育系插条生根的最佳处理为接种前用水进行预处理0.5h,外植体为完整插条,培养基为A1、A2,其生根率为97%,生根系数为5.3.