亚洲蝮属两种的种下分类研究(蛇亚目:蝮亚科)

对分布在辽东半岛和山东半岛的“黑眉”蝮蛇,依据Ｍａｙｒ（１９５３）两个样本间平均数比较的亚种划分的经典方法,进行分类学研究,辽东半岛半蛇岛蝮应为：蛇岛蝮千山亚种Ｇｌｏｙｄｉｕｓ ｓｈｅｄａｏｅｎｓｉｓ ｑｉａｎｓｈａｎｅｎｓｉｓ ｓｓｐ;ｎｏｖ．山东半岛产岩栖蝮为：岩栖蝮长岛亚种Ｇｌｏｙｄｉｕｓ ｓａｘａｔｉｌｉｓ ｃｈａｎｇｄａｏｅｎｓｉｓ ｓｓｐ;ｎｏｖ．。     

中介蝮蛇毒纤溶酶基因真核表达载体的构建及其功能的研究

目的:蛇毒纤维蛋白溶解酶能直接溶解纤维蛋白或纤维蛋白原,在心脑血管疾病的治疗方面具有潜在的应用价值。方法:采用双酶切技术从pMD18T-FLE I克隆载体上获得中介蝮蛇毒纤溶酶基因编码区,再将其亚克隆到真核表达载体pFASTBACHTa上,经转化、筛选和鉴定,获得重组真核表达质粒pFASTBACHTa-FLE。重组质粒经小鼠尾静脉快速注入小鼠体内,进行瞬时表达。结果:经SDS-PAGE和Western blot检测,表明在小鼠肝脏组织中有重组FLE蛋白表达。免疫组织化学检测证明该蛋白在小鼠肝脏组织中大量表达。纤维蛋白平板法鉴定该重组蛋白具有较高的纤溶活性,其活性呈现出剂量相关性和时间依赖性。因此为进一步对中介蝮蛇毒纤溶酶的应用奠定了坚实的基础。     

墨旱莲对4种蝮蛇毒引起的炎症和出血的影响

目的探讨墨旱莲提取液对短尾蝮蛇毒、蛇岛蝮蛇毒、白眉蝮蛇毒及尖吻蝮蛇毒所致的炎症和出血的影响。方法应用短尾蝮蛇毒、蛇岛蝮蛇毒、白眉蝮蛇毒及尖吻蝮蛇毒所致大鼠足跖肿胀的致炎模型,观察墨旱莲提取液对蛇毒所致大鼠足跖肿胀的影响。墨旱莲提取液分别与不同蛇毒混合,给小鼠腹部皮下注射,观察其对蛇毒引起的小鼠皮下出血的影响。结果墨旱莲提取液15g／kg连续2次灌胃给药,对短尾蝮蛇毒、蛇岛蝮蛇毒、白眉蝮蛇毒或尖吻蝮蛇毒所致大鼠足跖肿胀的急性炎症造模和短尾蝮蛇毒棉球肉芽肿的慢性炎症造模(20g／kg)均有明显的抑制作用,对这些蛇毒引起的小鼠皮下出血也能明显抑制。结论墨旱莲提取液对短尾蝮蛇毒、蛇岛蝮蛇毒、白眉蝮蛇毒及尖吻蝮蛇毒引起的炎症和出血均有明显的抑制作用。     

Austro-boreal disjunctions: a remarkable biogeographical pattern illustrated by Nan Ao Island, Guangdong, China

Aim To document disjunct, insular populations of animals separated from similar conspecifics distributed to the north, inland and upland, and compare this pattern with that historically described for northern Florida and the Florida Keys islands. Location Nan Ao Island is on the Tropic of Cancer 117° E in the South China Sea, eastern Guangdong Province, China. There are archipelagos of islets extending south‐east, and several coastal islets around Nan Ao. Methods Five field surveys were made in 1999–2001. Voucher specimens were tagged, catalogued, and deposited in major museum collections. The comparable Floridian disjunct pattern was researched and described in overview. Results Five species – a toad, two lizards and two snakes – collected on Nan Ao and three satellite islands were added to the two previously documented species – a lizard and a snake – that fit the austro‐boreal disjunct pattern. One lizard and one snake are minor range extensions of a hundred or few km. One snake's Nan Ao population is disjunct by about 450 km and one lizard's by about 550 km. The small, semi‐fossorial, short‐legged toad is the most notable, separated from the main area of its range by about 800 km. This is similar to the greatest distance between north Floridian and Keys disjunct populations. Main conclusions (1) Seven Nan Ao species of terrestrial vertebrates are isolated and disjunct from conspecifics found to the north, upland and inland; this compares to five species showing a similar north Florida to Florida Keys pattern. (2) The Nan Ao species have no conspecifics known from the intervening area whereas four of the five Floridian species have conspecific, but dissimilar, populations in the gap. (3) Nan Ao and its satellite islets were high points on a large coastal plain at glacial maximum. (4) Holocene sea level rise and concomitant global warming may have isolated the Nan Ao relicts that had continuous distributions at glacial maximum. (5) The intervening mainland conspecific populations may have been replaced by tropical species that moved north along the mainland but failed to colonize the islands. (6) If so, significant adaptation to a warmer climate must have occurred to enable survival of the Nan Ao relicts to the present.     

河北兴隆县雾灵山蝮蛇分类地位的探讨

河北兴隆县的蝮蛇分类问题,以往有国外学者进行过报道.国内文献记载河北省只分布有一种蝮蛇即短尾蝮.2006年实地考察和饲养观察研究发现,雾灵山的蝮蛇不是同一种,而是两种蝮蛇.它们的形态、分布和习性均不同,依照中国动物志爬行纲第三卷鉴定,分别为短尾蝮和中介蝮,从而恢复了河北省中介蝮分布的纪录.     

重组白眉蝮蛇去整合素定点突变对其生物活性的影响

RGD为存在于许多糖蛋白配体中的氨基酸序列,对整合素具有识别作用.此序列也发现于许多蛇毒去整合素分子中.采用基因克隆技术从大连产白眉蝮蛇的毒腺中克隆出的去整合素adinbitor是含73个氨基酸残基的去整合素,分子中含有12个半胱氨酸和RGD模体.实验证明,adinbitor作为去整合素的新成员,具有典型的抗ADP诱导的人血小板聚集作用和抗肿瘤血管新生作用.为了将adinbitor的这2种功能分开,采用PCR基因定点突变的方法,将其cDNA序列中RGD模体改变成KGD.重组adinbitor(KGD)在E.coli BL21得到表达,并通过His•Bind亲和层析予以纯化.实验发现,adinbitor对ADP诱导的人血小板聚集具有明显抑制作用,其IC50=85 nmol/L,明显优于adinbitor(RGD) （IC50=150 nmol/L）.然而,与adinbitor(KGD)相比,adinbitor(KGD)则丧失了对血管生成的抑制作用.结果说明,adinbitor(KGD)可作为专一的抗人血小板聚集药具有潜在的开发前景.     

A phylogeographic, demographic and historical analysis of the short-tailed pit viper (Gloydius brevicaudus): evidence for early divergence and late expansion during the Pleistocene

The impact of quaternary glaciation in eastern China on local fanua and flora has been a topic of considerable interest. We used mitochondrial DNA (mtDNA) sequence data and coalescent simulations to test two general biogeographic hypothesis related to the effects of Pleistocene climatic fluctuations for a widespread ophidian species (Gloydius brevicaudus) in eastern China and Korean Peninsula. The phylogenetic analysis revealed three major lineages, the southeast Coastal, Yangtze and North Lineages. The latter two are closely related and jointly form a continental lineage. Divergence dating and coalescent simulations indicate a Late Pliocene to Early Pleistocene divergence between lineages from the southeast coast and continental interior, followed by a mid-to-late Pleistocene divergence between lineages from the north and the middle-lower Yangtze Valley across East China, suggesting that all these lineages predated the last glacial maximum. An overlapping range between the two lineages within the continental lineage and a secondary contact associated with ecological transition zones on the margins of the North China Plain were also observed. These results show that vicariance patterns dominated the history of G. brevicaudus. Though the climatic events of the Pleistocene have had a marked effect on the historical distribution and intra-specific divergence of reptiles in China, coalescent and non-coalescent demographic analyses indicate that all lineages of G. brevicaudus seem not to have been adversely affected by glacial cycles during the Late Pleistocene, presumably because of an increase in the amount of climatically mild habitat in East Asia due to a decline in elevation and the development of monsoons since the Mid-End Pleistocene.     

影响蛇岛蝮蛇树栖分布相关因子的分析

目的了解影响蛇岛蝮蛇树栖分布的7个相关因子,探讨其间的关系。方法2006年10月12~15日,在蛇岛二沟南坡、北坡、西坡,测定不同年龄组蛇岛蝮蛇树栖环境的7个相关因子,进行比较分析。结果三种树栖环境中,老成体、成体、亚成体、幼体对因子的选择基本都有差异;不同年龄组的树栖高度与冠幅负相关不显著,与枝长(成体和亚成体中)负相关不显著,与其它正相关不显著;老成体和成体在林中和林缘,对树栖高度的选择差异显著,亚成体和幼体差异不显著;相同年龄组两两因子间线性关系明显。结论影响蛇岛蝮蛇树栖分布的因子具有一定的规律性。     

蛇岛蝮蛇消化道5-羟色胺细胞的形态与分布

采用ABC(avidin-biotin-peroxidase complex)免疫组织化学方法,观察蛇岛蝮蛇(Gloydius shedaoensis)消化道内5-羟色胺(5-HT)免疫阳性内分泌细胞的分布及形态.结果显示,5-羟色胺细胞从食管到直肠各段均有分布.细胞分布密度呈波浪式,其中胃贲门部分布密度最高(7.15±2.38),直肠部次之(4.55±3.14),食管部最低(1.2±0.71).5-HT阳性细胞广泛分布于消化道上皮细胞之间、上皮基部、腺泡上皮细胞之间以及固有膜内.形态多样,呈圆形、锥体形、梭形等.分析认为蛇岛蝮蛇消化道5-HT细胞具有内、外分泌两种作用途径,并且其密度分布可能与其食性、生存环境有关.