基于16S基因序列海南产白唇竹叶青蛇的分类地位

为研究海南产白唇竹叶青蛇分类地位,捕捉来自海南岛各地的12个白唇竹叶青蛇样本,并从GenBank下载65个竹叶青蛇的样本序列,以瘤鼻蝮蛇为外群,建立16S系统发育树。系统发育树显示,竹叶青蛇分为多个属的观点具有很大的合理性,白唇竹叶青蛇隶属于帝汶(岛屿)竹叶青蛇属。进一步验证了经典形态学和生物地理学的观点,海南岛产白唇竹叶青与其它地方产白唇竹叶青蛇不存在生殖隔离。     

四黄散外敷对竹叶青蛇咬伤肿胀的疗效观察

目的探讨四黄散外敷治疗竹叶青蛇咬伤肿胀的临床疗效。方法选择2017年10月~2020年4月我院收治的112例竹叶青蛇咬伤肿胀患者为研究对象,采用随机数字表法将患者分为观察组和对照组,每组56例。对照组采用常规蛇咬伤处理方案,包括清创、引流、注射抗蛇毒血清及对症治疗等。观察组在对照组治疗基础上采用四黄散外敷治疗。观察比较两组患者的有效止痛时间、肿胀开始消退时间、肿胀完全消退时间,并对两组患者治疗7d时的临床疗效进行比较。结果观察组在有效止痛时间、肿胀开始消退时间、肿胀完全消退时间方面均显著低于对照组,两组比较差异具有统计学意义(均P<0.05)。两组治疗后的总体疗效比较,观察组的治愈率为89.3%(50/56),对照组的治愈率为69.6%(39/56),两组比较差异有统计学意义(P<0.05)。结论四黄散外敷治疗竹叶青蛇咬伤肿胀的疗效确切,能有效减轻患者的疼痛感,促进肿胀消退,减轻炎症反应,值得临床应用。     

危型竹叶青蛇伤性DIC的救治

目的：通过比较单纯中医药治疗与抗蛇毒血清及中西医结合救治方法,观察竹叶青蛇伤性DIC患者的疗效。方法：用抗蛇毒血清、立止血及中西医结合的方法。从1996年8月至2002年9月,共收治危型竹叶青蛇伤性DIC患者25例。结果：25例均治愈。结论：抗蛇毒血清的早期应用是有效方法。抗五步蛇毒血清或抗蝮蛇毒血清和立止血合用,可以对竹叶青蛇毒的出血毒素和促凝毒性起较好的中和作用。     

Interisland Mutation of a Novel Phospholipase A<Subscript>2</Subscript> from <Emphasis Type="Italic">Trimeresurus flavoviridis</Emphasis> Venom and Evolution of Crotalinae Group II Phospholipases A<Subscript>2</Subscript>

Trimeresurus flavoviridis (Crotalinae) snakes inhabit the southwestern islands of Japan: Amami-Oshima, Tokunoshima, and Okinawa. Affinity and conventional chromatographies of Amami-Oshima T. flavoviridis venom led to isolation of a novel phospholipase A₂ (PLA₂). This protein was highly homologous (91%) in sequence to trimucrotoxin, a neurotoxic PLA₂, which had been isolated from T. mucrosquamatus (Taiwan) venom, and exhibited weak neurotoxicity. This protein was named PLA-N. Its LD₅₀ for mice was 1.34 µg/g, which is comparable to that of trimucrotoxin. The cDNA encoding PLA-N was isolated from both the Amami-Oshima and the Tokunoshima T. flavoviridis venom-gland cDNA libraries. Screening of the Okinawa T. flavoviridis venom-gland cDNA library with PLA-N cDNA led to isolation of the cDNA encoding one amino acid-substituted PLA-N homologue, named PLA-N(O), suggesting that interisland mutation occurred and that Okinawa island was separated from a former island prior to dissociation of Amami-Oshima and Tokunoshima islands. Construction of a phylogenetic tree of Crotalinae venom group II PLA₂s based on the amino acid sequences revealed that neurotoxic PLA₂s including PLA-N and PLA-N(O) form an independent cluster which is distant from other PLA₂ groups such as PLA₂ type, basic [Asp⁴⁹]PLA₂ type, and [Lys⁴⁹]PLA₂ type. Comparison of the nucleotide sequence of PLA-N cDNA with those of the cDNAs encoding other T. flavoviridis venom PLA₂s showed that they have evolved in an accelerated manner. However, when comparison was made within the cDNAs encoding Crotalinae venom neurotoxic PLA₂s, their evolutionary rates appear to be reduced to a level between accelerated evolution and neutral evolution. It is likely that ancestral genes of neurotoxic PLA₂s evolved in an accelerated manner until they had acquired neurotoxic function and since then they have evolved with less frequent mutation, possibly for functional conservation. The nucleotide sequences reported in this paper are available from the GenBank/EMBL/DDBJ databases under accession numbers AB102728 and AB102729.     

Maximizing information in systematic revisions: a combined molecular and morphological analysis of a cryptic green pitviper complex (Trimeresurus stejnegeri)

Although the integration of DNA information in taxonomy has been invaluable, logistical problems relating to sampling can seriously limit its applicability. Here we describe the analysis of a morphologically cryptic species complex, in which we maximize the information present by using both a DNA phylogeny and a multivariate morphometric approach. The green pitviper Trimeresurus stejnegeri s.l. is widespread in Asia, with a number of described subspecies (some of which are considered full species by some workers) and two new species that have recently been described from Thailand. The phylogeny indicates three clades, which can also be discerned in the principal component analyses of morphological variation. Combining molecular and morphological information permits evaluation of the taxonomic position of populations not represented in the phylogeny − in particular, the subspecies T. s. chenbihui and T. s. yunnanensis . We discuss nomenclatural issues raised by this analysis, although these cannot be fully resolved until the holotypes of these subspecies can be examined. Finally, we apply a molecular clock calibrated in New World pitvipers, and discuss some of the palaeoclimatic changes that might have impacted upon diversification in this group.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society, 2004, 82 , 219–235.     

Multiple causation of phylogeographical pattern as revealed by nested clade analysis of the bamboo viper (Trimeresurus stejnegeri) within Taiwan

In order to assess the utility of nested clade analysis, both standard phylogenetic algorithms and nested clade analysis were performed on a geographically widespread survey of mitochondrial DNA haplotypes of the bamboo viper, Trimeresurus stejnegeri, within Taiwan. Gross tree topologies were congruent for all analyses and indicated the presence of two geographically overlapping clades within Taiwan. The smaller lineage was restricted to the north and east coasts, whereas the larger lineage occupied all but the northern range of the species within Taiwan including the Pacific offshore populations of Green and Orchid Islands. The phylogeographical pattern supports the existence of at least one colonization event from the continent since the initial isolation of Taiwan from the mainland in the Pliocene. However, determining the exact number of colonization events was not possible due to the simultaneous vicariant forces of hypothesized continental landbridge connections and the occurrence of dramatic in situ orogenesis throughout the Pleistocene. Nested clade analysis provided multiple temporal and spatial population historical inferences that are not possible with standard analyses and therefore should become widely applied to future phylogeographical studies.     

Purpureotin: a novel di-dimeric C-type lectin-like protein from Trimeresurus purpureomaculatus venom is stabilized by noncovalent interactions

Purpureotin, a novel di-dimeric C-type lectin-like protein (CLP) from Trimeresurus purpureomaculatus, was purified and sequenced. While its native molecular mass was determined to be 63kDa, purpureotin showed a single band of 30kDa on nonreducing SDS-PAGE and two polypeptide chains (16.0 and 14.5kDa) under reducing condition. These results were subsequently confirmed by mass spectrometric analyses. Based on these results, we postulate that purpureotin is a dimer of the alpha,beta-heterodimer which is held together by noncovalent interactions. Molecular modeling studies indicate that a dimer of alpha,beta-heterodimers can be formed where the alpha chains are held together by electrostatic charges and beta chains via hydrophobic interactions. Functionally, purpureotin induced platelet aggregation without any cofactor in a dose-dependent manner. However, the platelet aggregation effect was blocked by echicetin. Therefore, purpureotin is assumed to be a GPIb-binding protein which binds to the same or a closely related GPIb site on platelets as echicetin.     

兔循环血中黄绿烙铁头蛇毒浓度与DIC出现关系研究

目的观察兔循环血中黄绿烙铁头蛇毒浓度与ＤＩＣ出现的关系。方法用抗烙铁头蛇毒的鼠多克隆抗体ＩｇＧ测定兔血中蛇毒浓度,以α２－ＰＩ、ＡＴ－Ⅲ活性和纤维蛋白原水平参数为ＤＩＣ指标。结果上述指标水平随血中蛇毒浓度的增加而降低,ＤＩＣ的发生与血中蛇毒浓度呈负相关系,高剂量的蛇毒可诱发ＤＩＣ。结论ＤＩＣ的发生取决于兔循环血中蛇毒的浓度。     

Annual survival rate and age structure of habu,Trimeresurus flavoviridis (Viperidae), on Minna island,Japan: Estimation from removal trapping

A population of a viperid snake,Trimeresurus flavoviridis, was studied over 10 years by removal trapping on a small subtropical island in Japan. The sex ratio of trapped individuals changed seasonally, but was not biased to either sex in the whole sample of 258 individuals. The age of each individual was estimated through the size structure and the age-size relationship. The minimum number of individuals at the beginning of the study was estimated through accumulating older individuals trapped in the subsequent years. By assuming an annual natural survival rate in the course of this accumulation, an age structure was simulated which led to calculate a resulted natural survival rate. The assumed rate of 0.62 fitted best to the resulted one. The annual trapped proportion estimated on the simulated absolute number of individuals was higher in older individuals than in younger ones with the overall mean of 16%.