A new compound,6-methylsulfinylthieno[3,2-g]-2,4(1H,3H)-pteridinedione,was isolated from the water extract of Whitmania pigra Whitman.Its structure was elucidated on the basis of NMR and ESIMS spectra.
The artificial breeding technology for juvenile of Whitmania pigra was introduced in the paper,including selection of sites and water quality,construction of spawning pool,hatching pool and escape proof facilities,key...The artificial breeding technology for juvenile of Whitmania pigra was introduced in the paper,including selection of sites and water quality,construction of spawning pool,hatching pool and escape proof facilities,key technology of leech selection,feeding,cocoon hatching,juvenile feeding and management.展开更多
Whitmania pigra has been used as a traditional Chinese medicine(TCM) for promoting blood circulation, alleviating blood coagulation, activating meridians and relieving stasis for several hundred years. However, the th...Whitmania pigra has been used as a traditional Chinese medicine(TCM) for promoting blood circulation, alleviating blood coagulation, activating meridians and relieving stasis for several hundred years. However, the therapeutic components of this species, especially proteins and peptides were poorly exploited. Until now only a few of them were obtained by using chromatographic isolation and purification. In recent decade, transcriptome techniques were rapidly developed, and have been used to fully reveal the functional components of many animal venoms. In the present study, the c DNA of the salivary gland of Whitmania pigra was sequenced by illumina and the transcriptome was assembled by using Trinity. The proteome were analysed by LC-MS/MS. Based on the data of the transcriptome and the proteome, a potential antiplatelet protein named pigrin was found. Pigrin was cloned and expressed using P. pastoris GS115. The antiplatelet and antithrombotic bioactivities of pigrin were tested by using aggregometer and the rat arterio-venous shunt thrombosis model, respectively. The bleeding time of pigrin was measured by a mice tail cutting method. The docking of pigrin and protease-activated receptor 1(PAR1) or collagen were conducted using the ZDOCK Server. Pigrin was able to selectively inhibit platelet aggregation stimulated by PAR1 agonist and collagen. Pigrin attenuated thrombotic formation in vivo in rat, while did not prolong bleeding time at its effective dosage. There are significant differences in the key residues participating in binding of Pigrin-Collagen complex from Pigrin-PAR1 complex. In conclusion, a novel PAR1 inhibitor pigrin was found from the leech Whitmania pigra. This study helped to elucidate the mechanism of the leech for the treatment of cardiovascular disorder.展开更多
Using the method of intracellular injection of horseradish peroxidase, the arborizations of several neurones with different functions in the segmental ganglia of the leech Whitmania pigra are investigated. Comparison ...Using the method of intracellular injection of horseradish peroxidase, the arborizations of several neurones with different functions in the segmental ganglia of the leech Whitmania pigra are investigated. Comparison is made in each kind of neurones in order to verify their typical pattern. Results show that the morphology of the axonal arbors of identified neurones has an accurate similarity within and among the individuals of the leech, and the axonal arborizations of the bilaterally identified neurones in the same ganglion have a stable symmetry. The possible reasons of the formation and its functional significance of the symmetrical nervous system are discussed.展开更多
An initial attempt reveals parameters of marked electric synapse on AP perceptive neuron of leech. The width of gap between marked AP and either neuro- or glio-process is 2-3 nm, the diameter of the hexagonal array of...An initial attempt reveals parameters of marked electric synapse on AP perceptive neuron of leech. The width of gap between marked AP and either neuro- or glio-process is 2-3 nm, the diameter of the hexagonal array of connexion is 8-9 nm with a central pole of 2-2.5 nm, the same as the structure of annular lamellar bodies internalized by AP-glia processes in the AP.展开更多
目的:采用分子模拟技术从宽体金线蛭来源多肽库中筛选抗凝活性多肽,结合分子对接初筛及分子动力学模拟复筛,最终通过抗凝实验验证得到宽体金线蛭抗凝活性多肽。方法:利用虚拟酶切技术对宽体金线蛭蛋白库进行酶切,得到宽体金线蛭多肽候选...目的:采用分子模拟技术从宽体金线蛭来源多肽库中筛选抗凝活性多肽,结合分子对接初筛及分子动力学模拟复筛,最终通过抗凝实验验证得到宽体金线蛭抗凝活性多肽。方法:利用虚拟酶切技术对宽体金线蛭蛋白库进行酶切,得到宽体金线蛭多肽候选库;选择凝血酶为凝血活性靶点,采用HPEPDOCK及Discovery Studio CDOCKER两种方法进行分子对接,并利用分子动力学模拟技术对分子对接初筛结果中对接结合能高的复合物进行动力学模拟复筛,通过MM-PBSA模块分析计算凝血酶-多肽复合物结合自由能;最后采用固相合成法化学合成复筛中稳定结合的宽体金线蛭多肽并进行体外抗凝活性测定。结果:经过虚拟酶切共得到3317条无毒多肽,通过分子对接进行初筛,结合分子动力学模拟复筛,得到1条结合凝血酶效果最好的目标抗凝肽,该多肽序列为QNTVGLDDFFSSYER,结合自由能为-427.506 kJ·mol^(-1)。凝血酶Arg233残基是该凝血酶-多肽复合物中介导结合相互作用的关键氨基酸。该宽体金线蛭抗凝肽在体外活性测定中确能延长凝血酶时间。结论:利用分子模拟技术有效筛选出1条宽体金线蛭抗凝活性肽,并经过抗凝实验验证,表明“模拟+实验”的活性肽筛选策略切实可行,该研究思路可为动物类中药物质基础研究提供参考。展开更多
基金supported by National Key Technologies R&D Program during Five-Year Plan Period from the Ministry of Science and Technology of the People's Republic of China(No.2006BAI06A04-04).
文摘A new compound,6-methylsulfinylthieno[3,2-g]-2,4(1H,3H)-pteridinedione,was isolated from the water extract of Whitmania pigra Whitman.Its structure was elucidated on the basis of NMR and ESIMS spectra.
基金Supported by Science and Technology Innovation Projects of Jurong City(2012018)
文摘The artificial breeding technology for juvenile of Whitmania pigra was introduced in the paper,including selection of sites and water quality,construction of spawning pool,hatching pool and escape proof facilities,key technology of leech selection,feeding,cocoon hatching,juvenile feeding and management.
基金supported by China Pharmaceutical University initiative program(No.CPU2018GY15)the National Natural Science Foundation for Young Scientists of China(No.81603647)
文摘Whitmania pigra has been used as a traditional Chinese medicine(TCM) for promoting blood circulation, alleviating blood coagulation, activating meridians and relieving stasis for several hundred years. However, the therapeutic components of this species, especially proteins and peptides were poorly exploited. Until now only a few of them were obtained by using chromatographic isolation and purification. In recent decade, transcriptome techniques were rapidly developed, and have been used to fully reveal the functional components of many animal venoms. In the present study, the c DNA of the salivary gland of Whitmania pigra was sequenced by illumina and the transcriptome was assembled by using Trinity. The proteome were analysed by LC-MS/MS. Based on the data of the transcriptome and the proteome, a potential antiplatelet protein named pigrin was found. Pigrin was cloned and expressed using P. pastoris GS115. The antiplatelet and antithrombotic bioactivities of pigrin were tested by using aggregometer and the rat arterio-venous shunt thrombosis model, respectively. The bleeding time of pigrin was measured by a mice tail cutting method. The docking of pigrin and protease-activated receptor 1(PAR1) or collagen were conducted using the ZDOCK Server. Pigrin was able to selectively inhibit platelet aggregation stimulated by PAR1 agonist and collagen. Pigrin attenuated thrombotic formation in vivo in rat, while did not prolong bleeding time at its effective dosage. There are significant differences in the key residues participating in binding of Pigrin-Collagen complex from Pigrin-PAR1 complex. In conclusion, a novel PAR1 inhibitor pigrin was found from the leech Whitmania pigra. This study helped to elucidate the mechanism of the leech for the treatment of cardiovascular disorder.
基金Project supported by the National Natural Science Foundation of China.
文摘Using the method of intracellular injection of horseradish peroxidase, the arborizations of several neurones with different functions in the segmental ganglia of the leech Whitmania pigra are investigated. Comparison is made in each kind of neurones in order to verify their typical pattern. Results show that the morphology of the axonal arbors of identified neurones has an accurate similarity within and among the individuals of the leech, and the axonal arborizations of the bilaterally identified neurones in the same ganglion have a stable symmetry. The possible reasons of the formation and its functional significance of the symmetrical nervous system are discussed.
文摘An initial attempt reveals parameters of marked electric synapse on AP perceptive neuron of leech. The width of gap between marked AP and either neuro- or glio-process is 2-3 nm, the diameter of the hexagonal array of connexion is 8-9 nm with a central pole of 2-2.5 nm, the same as the structure of annular lamellar bodies internalized by AP-glia processes in the AP.
基金Acknowledgement We thank the Mexican government (CONANP) for research permission to work at Palenque National Park granted to AE. Our research was supported by grants from the National Science Foundation (DDIG 0622386), the L.S.B. Leakey Foundation, the Department of Zoology, University of Wisconsin-Madison, and the Pittsburgh Zoo Conservation Fund. All animal care regulations and applicable national laws were followed. M. Marroquin, R. MacDonald, G. Campbell, J.M. Jose Dominguez, C. Hauglustaine, E Gimmie, and A. Herbert provided support in the field. C. Snowdon, T. Ziegler, J. Lambert, and two anonymous reviewers provided helpful comments on an earlier version of this manuscript.
文摘目的:采用分子模拟技术从宽体金线蛭来源多肽库中筛选抗凝活性多肽,结合分子对接初筛及分子动力学模拟复筛,最终通过抗凝实验验证得到宽体金线蛭抗凝活性多肽。方法:利用虚拟酶切技术对宽体金线蛭蛋白库进行酶切,得到宽体金线蛭多肽候选库;选择凝血酶为凝血活性靶点,采用HPEPDOCK及Discovery Studio CDOCKER两种方法进行分子对接,并利用分子动力学模拟技术对分子对接初筛结果中对接结合能高的复合物进行动力学模拟复筛,通过MM-PBSA模块分析计算凝血酶-多肽复合物结合自由能;最后采用固相合成法化学合成复筛中稳定结合的宽体金线蛭多肽并进行体外抗凝活性测定。结果:经过虚拟酶切共得到3317条无毒多肽,通过分子对接进行初筛,结合分子动力学模拟复筛,得到1条结合凝血酶效果最好的目标抗凝肽,该多肽序列为QNTVGLDDFFSSYER,结合自由能为-427.506 kJ·mol^(-1)。凝血酶Arg233残基是该凝血酶-多肽复合物中介导结合相互作用的关键氨基酸。该宽体金线蛭抗凝肽在体外活性测定中确能延长凝血酶时间。结论:利用分子模拟技术有效筛选出1条宽体金线蛭抗凝活性肽,并经过抗凝实验验证,表明“模拟+实验”的活性肽筛选策略切实可行,该研究思路可为动物类中药物质基础研究提供参考。