Triterpenoid saponins, new metalloprotease snake venom inhibitors isolated from Pentaclethra macroloba.

We report here the antiproteolytic and antihemorrhagic properties of triterpenoid saponin inhibitors, named macrolobin-A and B, from Pentaclethra macroloba, against Bothrops snake venoms. The inhibitors were able to neutralize the hemorrhagic, fibrin(ogen)olytic, and proteolytic activities of class P-I and P-III metalloproteases isolated from B. neuwiedi and B. jararacussu venoms. Clotting and fibrinogenolytic activities induced by snake venoms and isolated thrombin-like enzymes were partially inhibited. Furthermore, the potential use of these inhibitors to complement antivenom therapy as an alternative treatment and/or used as molecular models for development of new therapeutical agents in the treatment of snake bite envenomations needs to be evaluated in future studies.     

早期大剂量维生素C对重症急性胰腺炎大鼠核因子-κB的影响

目的:观察早期大剂量应用抗坏血酸(维生素C)对重症急性胰腺炎(SAP)大鼠的核因子-!B(NF-!B)的影响,研究其作用机制。方法:将72只SAP模型SD大鼠随机分成3组,每组各24只。A组:由大鼠股静脉滴注生理盐水5 ml/kg。B组:大鼠股静脉滴注Vit C 15 mg/kg加生理盐水至5 ml/kg。C组:由大鼠股静脉滴注Vit C 150 mg/kg加生理盐水至5 ml/kg。另取8只SD大鼠作为正常对照组。各组分别于8 h和24 h处死8只大鼠,采血测淀粉酶、脂肪酶、维生素C(PV-C)、超氧化物歧化酶(SOD)、TNF-αI、L-6。大鼠处死时分别取胰头组织3份,一份组织HE染色,行光镜检查,按Kusske的方法,对水肿、炎症、出血和坏死分别评分;一份制成超薄切片,行电镜检查;另一份SP法进行免疫组化染色,检测NF-!B的表达。每组另外8只大鼠观察3 d内存活情况,计算3 d成活率。结果:各组大鼠3 d内的生存率为正常对照组100%(8/8),A组0%(0/8),B组12.5%(1/8),C组50%(4/8),C组的3 d生存率显著高于其他两组(P<0.05)。各组的4项病理学评分均高于正常对照组(P<0.01),C组的4项病理学评分均低于A、B组(P<0.05)。透射电镜检查示C组中分泌颗粒较少,其包膜完整、内质网轻度肿胀、线粒体清晰,未见大片坏死。SAP大鼠体内淀粉酶、脂肪酶、细胞因子TNF-α和IL-6的水平明显增高,血清SOD和P-VC降低,胰腺组织中NF-!B活化阳性胰腺细胞数明显增多。C组的血清淀粉酶和脂肪酶低于A、B组(P<0.05),SOD和P-VC水平高于A、B组(P<0.05),血清TNF-αI、L-6水平低于A、B组(P<0.05),胰腺组织NF-!B活化水平低于A、B组(P<0.05)。结论:早期大剂量应用Vit C有助于及时提高SAP大鼠的P-VC、E-SOD水平,降低体内淀粉酶、脂肪酶、TNF-αI、L-6水平,其作用机制可能与大剂量Vit C抑制SAP大鼠体内NF-!B活化、在整体水平上抑制细胞因子基因表达、有助于机体免受自由基和过量细胞因子的损伤及减轻胰腺组织的病理性改变等因素有关。     

Immunocytochemical distribution of met-enkephalin-Arg6-Gly7-Leu8 in the rat lower brainstem

The distribution of methionine-enkephalin-Arg6-Gly7-Leu8, a unique peptide derived from proenkephalin A in the rat brainstem, was studied immunocytochemically by using a highly specific antiserum to this octapeptide sequence. Immunoreactive perikarya with various shapes and sizes were detected in many regions of the rat brainstem. Dense accumulation of immunoreactive perikarya and fibers was seen in the nuclei associated with special sensory and visceral functions, such as the interpeduncular nucleus, the parabrachial nucleus, the nucleus of the solitary tract, and the nucleus of the spinal tract of the trigeminal nerve. Clusters of methionine-enkephalin-Arg6-Gly7-Leu8-like immunoreactive perikarya and fibers were observed in certain areas considered to play a role in nociception and analgesia, such as the central gray of the midbrain central gray and the raphe magnus nucleus. Some methionine-enkephalin-Arg6-Gly7-Leu8-like immunoreactive perikarya were distributed in the lateral reticular nucleus, the nucleus of the solitary tract, and the raphe magnus nucleus, where monoaminergic neurons were also detected. In addition to the previously reported enkephalinergic cells, we found many methionine-enkephalin-Arg6-Gly7-Leu8 containing neurons; the rostral and caudal linear nucleus of raphe, the median raphe nucleus, entire length of the raphe magnus nucleus, the medial longitudinal fasciculus, the cuneate nucleus, the external cuneate nucleus, the gracile nucleus, and the area postrema. The wide distribution of this octapeptide-like immunoreactivity reflected neurons expressing the preproenkephalin A gene distributed more widely than previously reported and that innervated many regions.     

The participation of adenosine receptors in the adenosine 5''-triphosphate-induced relaxation in the isolated rabbit corpus cavernosum penis

AIM: To investigate the participation of adenosine receptors in the adenosine 5'-triphosphate (ATP)-induced relaxation in the corpus cavernosum penis (CCP) of rabbits. METHODS: The ATP-induced relaxation was assessed on the noradrenaline precontracted CCP of rabbits in the presence and absence of 8-(3-chlorostyryl)caffeine (CSC); an adenosine A(2A) receptor antagonist; alloxazine and MRS1754; adenosine A(2B) receptor antagonists; and ARL67156, an inhibitor of ecto-nucleoside triphosphate diphosphohydrolases. RESULTS: Adenosine and ATP relaxed the noradrenaline precontracted CCP of rabbits in a concentration-dependent manner. The adenosine- and ATP-induced relaxations were suppressed by alloxazine and MRS1754, but not by 8-(3-chlorostyryl)caffeine. ARL67156 potentiated the ATP-induced relaxation but not the adenosine-induced one. MRS1754 suppressed the ATP-induced relaxation potentiated by ARL67156. CONCLUSIONS: The above results suggest that, in the CCP of rabbits, the adenosine receptor mediating adenosine-induced relaxation is of the A(2B) receptor and the ATP directly causes relaxation through the A(2B) receptor on the CCP.     

云南省600例0~5岁儿童维生素A缺乏调查

目的:了解云南省城乡0~5岁儿童维生素A营养状况及影响因素。方法:省、地、县随机整群抽取600例儿童,采用微量荧光光度法检测VitA含量。结果:VitA缺乏发生率16·17%,地、县明显高于省会城市;0岁儿童明显高于其他年龄组;腹泻儿童明显高于正常儿童;VitA缺乏的影响因素:近1周内进食鸡蛋、奶类制品、鱼虾类、肝类、黄绿色蔬菜、鱼肝油等食物有利于维生素A的吸收,急性呼吸道感染、发烧与维生素A缺乏发生率差异不明显。结论:维生素A缺乏发生率城市明显低于农村地、县两级,说明维生素A缺乏防治工作重点在农村,特别是0岁儿童,防治的主要措施是采取合理膳食,均衡营养,控制腹泻流行。     

南山区与某社区常住人口6年恶性肿瘤发病调查

目的：了解某社区常住人口恶性肿瘤平均年发病率是否高于周边人群，重点是白血病平均年发病率。方法：采用回顾性调查法及个案调查表收集该社区1998年6月～2004年9月常住人口中的确诊恶性肿瘤病人22例，将该社区恶性肿瘤平均年发病率与南山区在该时段恶性肿瘤平均的发病率进行比较。结果：1998年6月～2004年9月南山区常住人口恶性肿瘤平均年发病66．1／10万，某社区常住人口恶性肿瘤平均年发病率29．9／10万，两地差异有统计学意义（F=0．0001，P〈0．05）；南山区常住人口白血病平均年发病率2．79／10万，某社区常住人口白血病平均年发病率4．07／10万，两者无统计学意义差异（F=0．467，P〉0．05）。结论：该社区常住人口恶性肿瘤及白血病平均年发病水平低于南山区全区水平。     

大鼠维生素D受体蛋白表达载体的构建及遗传性高钙尿性结石大鼠十二指肠维生素D受体cDNA序列分析

目的构建大鼠维生素D受体（VDR）蛋白表达载体，测定并分析遗传性高钙尿性结石（GHS）大鼠VDRcDNA序列。方法采用半定量逆转录-聚合酶链反应（RT—PCR）方法扩增含VDR蛋白编码基因序列，将扩增产物克隆至真核表达载体pcDNA3．1／Zero（＋），对5只GHS大鼠和4只正常尿钙对照（NC）大鼠十二指肠VDRcDNA序列进行测序分析。结果琼脂糖凝胶电泳示PCR扩增产物碱基数目与目的片段大小一致。对重组质粒的分析表明，插入片段的序列与发表的VDR基因编码序列相同。5只GHS鼠和4只NC鼠肠VDR cDNA序列均有3个相同的位点不同于已公布的大鼠肠VDR cDNA序列：在256bp位点：C取代G；569bp位点：G代替A；1658位点：A替代G。另外，GHS鼠在1795bp位点发生改变，G取代A，而NC鼠则无改变。结论大鼠维生素D受体蛋白编码序列被成功地克隆至表达载pcDNA3．1／Zero（＋）上。GHS大鼠基因编码区的序列组成同NC大鼠相一致，但GHS鼠1795bp位点的碱基改变未见于NC鼠。     

A drug interaction study between ticlopidine and cyclosporin in heart transplant recipients

Objectives: Previous uncontrolled studies have suggested an interaction between ticlopidine, a major antiplatelet agent, and cyclosporin in heart- and kidney-transplant recipients. The aims of this study were to examine in a randomised, double-blind fashion, the possible interaction between cyclosporin A and ticlopidine (250 mg per day) and the tolerability of this combination in heart-transplant recipients. Methods: Twenty heart-transplant recipients were randomised into either a treated or a placebo group. Blood samples were drawn for time-course evaluation of cyclosporin blood levels over a period of 12 h, following the morning intake of cyclosporin and, for platelet aggregation studies, before and after 14 days of ticlopidine administration. Twenty four-hour urine samples were collected for 6-β-hydroxycortisol measurements, before and after 14 days of ticlopidine. Results: Although given at half the recommended daily dosage, ticlopidine significantly reduced platelet aggregation. Pharmacokinetic parameters indicate that the bioavailability of cyclosporin A was not significantly modified by ticlopidine. However, one patient in the ticlopidine group was withdrawn because of a major fall in cyclosporin blood level within 3 days of treatment. Urinary excretion of 6-β-hydroxycortisol was augmented after treatment in the ticlopidine group compared with the placebo group, suggesting that induction of drug metabolism might have occurred. Data also show quite a large intra-individual variability in cyclosporin bioavailability in the placebo group, suggesting that poor absorption of the drug formulation and/or poor compliance might have contributed to the decreased cyclosporin blood levels in the patient withdrawn from this study and in previous uncontrolled studies. Conclusion: Cyclosporin bioavailability was not clearly modified by a half dosage of ticlopidine in this study. We, however, recommend closely monitoring cyclosporin blood levels when prescribing ticlopidine. Further studies will be needed with new formulations of cyclosporin or when using the full dosage of ticlopidine. Received: 20 July 1996 / Accepted in revised form: 12 February 1997     

特异性荧光偏振免疫法监测521例肾移植后环孢素A全血浓度3275次

目的通过监测肾移植后病人环孢素A(CsA)全血浓度 ,提出CsA在三联免疫抑制用药方案中的理想治疗窗。方法用特异性荧光偏振免疫法测定CsA全血浓度 ,对521例病人监测3275次 ,按术后时间及临床表现分组比较。结果肾移植后<1 ,、1～3、3～6、6～12个月、1～2和>2年的CsA全血谷浓度的理想治疗窗应分别为250～450、200～400、150～300、100～250、100～200和100～180μg/L。结论CsA全血浓度在上述范围内 ,中毒反应和排异反应明显减少     

Structural Characterization of Variant Forms of Arylsulfatase A that Associate with Alcoholism

Several electrophoretic forms of human platelet arylsulfatase A (ASA), including variant type III_a and normal type IV_a, have been identified by nondenaturing polyacrylamide gel electrophoresis. An alcoholic population that we have analyzed is enriched in variant type III_a compared with nonalcoholic psychiatric and normal controls. Individuals with the III_a enzyme possess greatly reduced levels of ASA activity. To understand further the structural basis for the differences and their potential biological consequences, the nature of the ASA variant expressed by fibroblasts from different individuals was explored. The electrophoretic patterns of fibroblast ASA from the III_a and IV_a individuals differ in degree of phosphorylation. Furthermore, fibroblast ASA from III_a individuals lacks an N -linked glycan found in ASA from IV_a individuals. In addition, differences in peptide and/or posttranslational modification unrelated to the N -linked carbohydrate or phosphorylation exist between the fibroblast ASA from III_a and IV_a individuals. The finding that both fibroblasts and platelets exhibit related electrophoretic isoform patterns characteristic of the donor's ASA type allows for the use of fibroblasts to study the impact of ethanol on the metabolism of cells possessing different ASA types.