Purification and Characterization of Jerdonitin, a Non-hemorrhagic Metalloproteinase from Trimeresurus jerdonii Venom

Previously, we have purified Jerdonitin from Trimeresurusjerdonii venom. Compared with other P-Ⅱ class snake venom metalloproteinases （SVMPs）, Jerdonitin has a primary structure comprising metalloproteinase and disintegrin domains. However, no hemorrhagic and fibrinogenolytic activities were detected for Jerdonitin. We thought that organic buffer of high performance liquid charamatography （HPLC） might affect its enzymatic activity. In this study, we purified Jerdonitin by another procedure excluding the HPLC. It was homogenous as judged by SDS-PAGE and had an apparent molecular weight of 36 kDa under non-reducing conditions and 38 kDa under reducing conditions, respectively. Like other typical metalloproteinases, Jerdonitin preferentially degraded alpha-chain of human fibrinogen and this fibrinogenolytic activity was completely inhibited by EDTA, but not by PMSF. It was interesting that Jerdonitin did not induce hemorrhage after intradermal injection in mice.

A New Species of the Genus Trimeresurus from Southwest China(Squamata:Viperidae)

Species from the Trimeresurus popeiorum complex(Subgenus: Popeia) is a very complex group. T. popeiorum is the only Popeia species known from China. During the past two years, five adult Popeia specimens(4 males, 1 female) were collected from Yingjiang County, Southern Yunnan, China. Molecular, morphological and ecological data show distinct differences from known species, herein we describe these specimens as a new species Trimeresurus yingjiangensis sp. nov Chen, Ding, Shi and Zhang, 2018. Morphologically, the new species distinct from other Popeia species by a combination of following characters:(1) dorsal body olive drab,without cross bands on the scales;(2) a conspicuous bicolor ventrolateral stripe present on each side of males, first row of dorsal scales firebrick with a white ellipse dot on posterior upper part in male, these strips absent in females;(3) eyes firebrick in both gender;(4) suboculars separated from 3rd upper labial by one scale on each side;(5) ventrals 164–168(n = 5);(6) MSR 21.

A New Species of Trimeresurus Lacépède, 1804(Squamata: Viperidae) from Southwestern China, Vietnam, Thailand and Myanmar

The pit vipers of the genus Trimeresurus Lacépède, 1804 is one of the largest groups of Asian snakes, distributed from India to China and Indonesia. Recent surveys in Jiangcheng and Simao, Yunnan Province, China resulted in a new species previously allocated to T. albolabris. Combining morphological and molecular data, we describe it as Trimeresurus guoi sp. nov. The new species morphologically differs from T. albolabris in the yellow green ventral color;an indistinct ventrolateral line;the a bsence of a postocular stripe;the firebrickred iris;a dark red stripe on dorsal tail;hemipenes with relatively weak sparse papillae, reaching 23 rd subcaudal when unextruded. Molecularly, the new species forms a clearly divergent lineage(BPP 1.00/UFB 100). Uncorrected pairwise distances of mitochondrial gene Cyt b between the new species and other known species of the subgenus Trimeresurus range from 0.052(T. albolabris) to 0.071(T. insularis).

Changes in hippocampal histamine receptors in rats after treatment with Trimeresurus albolabris venom

BACKGROUND： It has been demonstrated that histamine and its receptors in the hippocampus play an important role in memory and/or learning behaviors.OBJECTIVE： To investigate the expression levels of the histamine receptor gene and protein in the hippocampi of rats prior to and after administration of Trimeresurus albolabris venom using reverse transcription-polymerase chain reaction （RT-PCR） and Western blot techniques. DESIGN, TIME AND SETTING： A controlled observation based on cellular protein level was performed in the College of Life Sciences, Chongqing Normal University between March 2005 and April 2007. MATERIALS： Eighty adult male Sprague-Dawley rats were provided by the Laboratory Animal Center of the Third Military Medical University of Chinese PLA. The lyophilized powder of Trimeresurus albolabris venom was collected from Jin-Hu-Shan in Chongqing, China. METHODS： Twenty rats were randomly and evenly divided into an experimental group and a control group The experimental group was subcutaneously injected with 0.65 mg/mL Trimeresurus albolabris venom, 0.5 mL for each rat. The control group was subcutaneously injected with an equal amount of 0.9% physiological saline. Prior to and after injection, rats from these two groups were placed in the Morris Water Maze for recording of path length and escape latency. The remaining 60 rats were randomly allocated to another experimental group （n = 50） and another control group （n = 10）. Rats were correspondingly injected as described above. At different time points （0.1, 0.5, 1, 2, and 3 hours after injection）, rats were decapitated and bilateral hippocampal tissues were dissociated （approximately 100 mg for each sample）. Then, the acquired hippocampal tissue was immediately preserved at -70 ℃ for subsequent experiments. MAIN OUTCOME MEASURES： （1） The levels of histamine receptor （including H1R, H2R, and H3R） mRNA and protein in the hippocampi of rats were measured prior to and after injection of Trimeresurus albolabris venom using RT-PCR and Western Blot techniques. （2） Escape latency （namely, time to reach a platform） and path length were examined by Morris Water Maze testing. RESULTS： All 80 rats were included in the final analysis. In the experimental group, the level of mRNA for H3R receptor in rat hippocampi was just slightly changed, but the level of H3R receptor protein was significantly down-regulated compared with that in the control group （P 〈 0.05）. Both mRNA and protein levels for H1R receptor were initially downregulated and then recovered to normal levels. Expression of H2R receptor mRNA was initially upregulated, then downregulated, and finally restored to the control level. The level of H2R receptor protein showed a tendency for downregulation. In the Morris Water Maze testing, escape latency and path length were significantly longer in the experimental group than in the control group （P 〈 0.05）. CONCLUSION： Within three hours of injection with Trimeresurus albolabris venom, mRNA and protein levels of most histamine receptors in rat hippocampi were downregulated. Such changes possibly contribute to an impairment of memory and/or learning behaviors in rats following injection of Trimeresurus albolabris venom.

Purification and characterization of an arginine ester hydrolase from the venom of Trimeresurus mucrosqumatus in Hunan province of China

Objective: To study the physical and chemical properties of an arginine ester hydrolase from the venom of Trimeresurus mucrosqumatus in Hunan province of China.Methods:The arginine ester hydrolase(AEH) was isolated from the venom of Chinese Trimeresurus mucrosqumatus by a combination of ion-exchange chromatography on DEAE-Sephadex A-50, CM-Sepharose Cl-6B and gel filtration on Sephadex G-100.Results: The purified protein named TM-AEH,a glycoprotein with carbohydrate content of 0.5% neutral hexose and 0.75% sialic acid,a relative molecular mass of 29.0 kDa,and an isoelectric point(pI) of 5.2. It shares with an extinction coefficient(E 0.1%/cm) of 1.332 at 280 nm,consisted of 225 amino acid residues,and migrated as a band under reduced or non-reduced condition in basic PAGE.TM-AEH was a highly thermostable protein and was stable to pH changes between 5 and 9.The optimum temperature and optimum pH were 55℃ and 8.4 for its catalytic activity respectively,which was inhibited by Fe 3+ and Cu 2+.Conclusion:This protein can exhibit higher BAEE-hydrolysing activity and fibrinogenolytic activity as compared to that of whole venom.

The Study on Taxonomic Status of Trimeresurus stejnegeri chenbihuii Based on Scale Counting Method

[Objectives] This study was conducted to further investigate the taxonomic status of Hainan population of Trimeresurus stejnegeri from the morphological point of view.[Methods]The difference coefficients between different populations were compared using the 75% law,and the relationship between scales and latitudes was analyzed.[Results] The scales(abdominal and subcaudal) of 325 Trimeresurus individuals were counted according to China Animal Fauna,including156 T.stejnegeri individuals.Some difference coefficients between the Hainan population and others were greater than 1.28,and there was no correlation between the number of scales and latitude.It conforms to subclassification criteria.[Conclusions]The view about the subspecies status of T.stejnegeri chenbihuii should be supported.

The Exploration of Trimeresurus Divided to Multiple Genera Based on Molecular Systematics

According to the contradictions about classification on Trimeresurus between ＂China＇s Fauna＂ and international database, the rationality about Trimeresurus divided to multiple genera based on molecular systematies was discussed. The genetic distance was calculated by sequencing 12S gene of the original Trimeresurus snakes, including 80 individuals of 33 species ; and the molecular phylogenetie tree was established by taking Hypnale hypnale as the outgroup. The results showed that the topological structures of ME tree and ML tree were basically consistent, and all of the species in the molecular trees were divided into 8 branches; and the difference of T. albolabris between Hainan and other locations is not obvious. The viewpoint that original Trimeresurus genera subdivided into 8 genera was more rational, and new authoritative reference books are expected to be published in China.

广义竹叶青蛇属的系统学研究新进展

2004年以来,广义竹叶青蛇的系统学研究取得了较大的进展。本文对这近一年来的最新进展进行概括和报道。

江苏省鸟类一新记录──黄爪隼

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