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
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
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.
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
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
目的:利用分子模拟技术筛选以凝血因子Ⅺa(FⅪa)为靶点的宽体金线蛭抗凝多肽,并通过实验验证其抗凝活性。方法:采用“模拟+实验”的活性肽筛选策略,以FⅪa为抗凝活性靶点,首先采用HPEPDOCK和Discovery Studio ZDOCK 2种分子对接技术进...目的:利用分子模拟技术筛选以凝血因子Ⅺa(FⅪa)为靶点的宽体金线蛭抗凝多肽,并通过实验验证其抗凝活性。方法:采用“模拟+实验”的活性肽筛选策略,以FⅪa为抗凝活性靶点,首先采用HPEPDOCK和Discovery Studio ZDOCK 2种分子对接技术进行初步筛选,随后运用分子动力学模拟方法对结合能较高的FⅪa-多肽复合物进行动力学模拟复筛。通过MM/PBSA模块计算复合物的结合自由能,筛选出稳定结合的宽体金线蛭多肽。最后,采用固相合成法化学合成目标多肽,并进行体外抗凝活性测定以验证其生物活性。结果:筛选出1条与FⅪa结合效果最佳的抗凝肽,其氨基酸序列为VILTNPSGTLTEAIVK。体外活性实验结果表明,该宽体金线蛭来源的抗凝肽能够显著延长凝血时间。通过发色底物法进一步验证,证实该多肽能够与FⅪa结合并显著抑制其对发色底物s-2366的酶切活性。结论:筛选出1条以FⅪa为靶点的宽体金线蛭抗凝多肽,其能够通过特异性酶切FⅪa显著延长凝血时间,展现出良好的抗凝活性。该研究验证了“模拟+实验”筛选策略的有效性,为开发新型抗凝药物提供了潜在的候选多肽。展开更多
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 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.
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.
文摘目的:利用分子模拟技术筛选以凝血因子Ⅺa(FⅪa)为靶点的宽体金线蛭抗凝多肽,并通过实验验证其抗凝活性。方法:采用“模拟+实验”的活性肽筛选策略,以FⅪa为抗凝活性靶点,首先采用HPEPDOCK和Discovery Studio ZDOCK 2种分子对接技术进行初步筛选,随后运用分子动力学模拟方法对结合能较高的FⅪa-多肽复合物进行动力学模拟复筛。通过MM/PBSA模块计算复合物的结合自由能,筛选出稳定结合的宽体金线蛭多肽。最后,采用固相合成法化学合成目标多肽,并进行体外抗凝活性测定以验证其生物活性。结果:筛选出1条与FⅪa结合效果最佳的抗凝肽,其氨基酸序列为VILTNPSGTLTEAIVK。体外活性实验结果表明,该宽体金线蛭来源的抗凝肽能够显著延长凝血时间。通过发色底物法进一步验证,证实该多肽能够与FⅪa结合并显著抑制其对发色底物s-2366的酶切活性。结论:筛选出1条以FⅪa为靶点的宽体金线蛭抗凝多肽,其能够通过特异性酶切FⅪa显著延长凝血时间,展现出良好的抗凝活性。该研究验证了“模拟+实验”筛选策略的有效性,为开发新型抗凝药物提供了潜在的候选多肽。
