The cDNA encoding a variant of α bungarotoxin was cloned from the venom glands of Bungarus multicinctus by RT PCR.The deduced protein precursor contained a 21 amino acid signal peptide and a following 74 amino ...The cDNA encoding a variant of α bungarotoxin was cloned from the venom glands of Bungarus multicinctus by RT PCR.The deduced protein precursor contained a 21 amino acid signal peptide and a following 74 amino acid mature protein.The signal peptide is very similar to those of short chain neurotoxins,κ neurotoxins and cardiotoxins.The amino acid sequence of the mature protein is identical to α bungarotoxin (V31),a minor variant of α bungarotoxin identified by protein sequencing technique.Furthermore,the cDNA encoding the deletion precursor of α bungarotoxin was also cloned.By use of pMAL p2,the variant was overexpressed in E coli as a soluble fusion protein and purified by sepharose 6B amylose affinity chromatography,which was confirmed by western blotting with the antisera agai nst α bungarotoxin.The recombinant variant was achieved after digestion by factor X a.It displayed about 1/6 in vivo toxicity of natural α bungarotoxin.The successful cloning and functional expression of α bungarotoxin provided a basis for the future study of structure function of long neurotoxins.展开更多
Aim To investigate the anticancer activity of two new cytotoxins from thevenom of Agkistrodon acutus. Methods The venom was isolated by FPLC column chromatography consistingof DEAE Sepharose FF and Source 30S. The cyt...Aim To investigate the anticancer activity of two new cytotoxins from thevenom of Agkistrodon acutus. Methods The venom was isolated by FPLC column chromatography consistingof DEAE Sepharose FF and Source 30S. The cytotoxic activity on tumor cells was detected by MITmethod. Purity and molecular weight were determined by SDS-PAGE (silver staining). Their stabilitiesto temperature and pH were also detected. Results Two pure cytotoxins named ACTX-6 and ACTX-8 wereobtained. Their molecular weights are 98 kDa and 27 kDa, respectively. ACTX-6 consists of twosubunits bonded together by disulfide bonds. Conclusion ACTX-6 and ATCX-8 have highest inhibitoryactivity on lung cancer cell A549. ACTX-6 is stable to heat while ACTX-8 not. ACTX-6 is stablebetween pH 7-9 and ACTX-8 between pH 6 - 9.展开更多
[Objective] This study aimed to obtain recombinant alpha-bungarotoxin （a-BG-0 gene fusion protein with biological activity and investiagte its fusion expression. [Method] The plasmid pGEX-a-BGT was transformed into E...[Objective] This study aimed to obtain recombinant alpha-bungarotoxin （a-BG-0 gene fusion protein with biological activity and investiagte its fusion expression. [Method] The plasmid pGEX-a-BGT was transformed into E coil BL21 （DE3） and BL21 （DE3） plysS host bacteria to identify the optimal engineering strain. Fusion expression of the optimal engineering strain was induced, in order to optimize the induced expression conditions of the soluble fusion protein. [Result] JP-a-BGT was identified as the optimal engineering strain, which could express fusion protein after induced by IPTG. The optimal induced expression conditions of the soluble fusion protein were investigatect JP-a-BGT was incubated at 37 ℃ for 2.5 h and induced with 0.50 mmol4. IPTG for 4 h at 22 ℃, and the expression level of the soluble fusion protein reached 18.42%. [Conclusion] This study laid a solid foundation for the subsequent purification of fusion proteins and the separation and purification of a-BGT.展开更多
Aim To study chemical constituents from Ampelopsis grossedentata. Methods Separation and purification were performed by using silica gel, polyamide, reverse-phase silica gel, Sephadex LH-20 column chromatographic tech...Aim To study chemical constituents from Ampelopsis grossedentata. Methods Separation and purification were performed by using silica gel, polyamide, reverse-phase silica gel, Sephadex LH-20 column chromatographic techniques and silica gel PTLC. Structures were determined by means of physicochemical properties and spectral analysis. Results Four flavonoids were separated and identified from Ampelopsis grossedentata including dihydromyricetin (1), myricetin (2), myricitrin (3), and myricetin-3-O-β-D-galactopy...展开更多
Total mRNA was extracted from a venom gland of snake Daboia russellii siamensis following the manufacturer＇s protocol of the PolyATtract System 1000 kit purchased from Promega Biotech. cDNAs encoding C-type lectins w...Total mRNA was extracted from a venom gland of snake Daboia russellii siamensis following the manufacturer＇s protocol of the PolyATtract System 1000 kit purchased from Promega Biotech. cDNAs encoding C-type lectins were amplified by RT-PCR and subcloned into a pMD18-T vector. Fourteen positive clones were selected for nucleotide sequencing and seven cDNAs encoding various snake venom C-type lectin-like protein precursors, designated as DRS-L1, DRS-L2, DRS-L3, DRS-L4 DRS-L5, DRS-L6 and DRS-L7, were obtained. Amino acid sequences of these proteins were deduced and each contains a carbohydrate recognition domain. Of all the deduced protein sequences, only DRS-L1 seemed to represent a closer sequence similarity to α subunits of other known snake venom C-type lectin-like proteins using the BLAST program. Homology comparison combined with analysis of cysteine position indicate that DRS-L1 and DRS-L2 are probably the light chain LC2 and LC1 of factor X activator from Daboia russellii siamensis venom, respectively. DRS-L3 and DRS-L4 might be the β subunits of higher molecular weight C-type lectin-like proteins while DRS-L5 and DRS-L6 might be β subunits of lower molecular weight C-type lectin-like proteins. DRS-L7 might be the β subunit of a platelet membrane glycoprotein Ib-binding protein similar to echicetin.展开更多
A new angular furanocoumarin, 2′ acetylangelicin (8), along with oroselone (1), columbianadin (3), bergapten (4), O acetyl columbianetin (5), cniforin A (6), edultin (7) and β sitosterol (2), was isolated fro...A new angular furanocoumarin, 2′ acetylangelicin (8), along with oroselone (1), columbianadin (3), bergapten (4), O acetyl columbianetin (5), cniforin A (6), edultin (7) and β sitosterol (2), was isolated from the fruit of Cnidium monnieri. The structures were elucidated by spectral methods. Compound 1 was isolated from this plant for the first time.展开更多
Objective： To investigate the effects of rAdinbitor on the adhesion and proliferation of human hepatoma cell strain 8MMC-7721. Methods： Cell adhesion assay was used to observe the effect of rAdinbitor on the adhesio...Objective： To investigate the effects of rAdinbitor on the adhesion and proliferation of human hepatoma cell strain 8MMC-7721. Methods： Cell adhesion assay was used to observe the effect of rAdinbitor on the adhesion of 8MMC-7721 cells to fibronectin （FN）. Crystal violet staining was performed to detect the influence of rAdinbitor on the adhesion of 8MMC- 7721 cells. MTT assay was employed to detect the inhibitory effects of different concentration of rAdinbitor on the proliferation of 8MMC-7721 cells. The morphologic changes of the control 8MMC-7721 cells and the apoptotic cells induced by 200μg/mL rAdinbitor for 36 h were observed under light microscope after HE staining. Flow cytometry analysis was applied to determine the apoptosis rate of 8MMC-7721 cells. Results： （1） FN promoted the adhesion of human hepatoma cell strain 8MMC-7721 in a dose-dependent manner. （2） rAdinbitor could dose-dependently inhibit the adhesion of SMMC-7721 cells to FN. The higher the concentration was, the stronger the inhibition was. There was significant difference among the groups （P 〈 0.05）. （3） rAdinbitor had a strong inhibition on the proliferation of 8MMC-7721 cells and showed a dose-dependent manner （P 〈 0.05）. After a 48 h exposure, the IC50 value of rAdinbitor was 177.83 μg/mL. （4） After exposure of 8MMC-7721 cells to 200μg/mL rAdinbitor for 36 h, the early morphologic changes appeared and the apoptosis rate was 20.68%, significantly higher than that of the control group （2.38%, P 〈 0.05）. Conclusion： rAdinbitor can dose-dependently inhibit the 8MMC-7721 cells adhesion to FN, and can inhibit the proliferation in dose-dependent manner and promote their apoptosis.展开更多
Peridotites from the southern Mariana forearc were sampled on the landward trench slope of the Izu-Bonin-Mariana (IBM) subduction zone by dredging.These mantle wedge peridotites underwent hydration by fluid derived fr...Peridotites from the southern Mariana forearc were sampled on the landward trench slope of the Izu-Bonin-Mariana (IBM) subduction zone by dredging.These mantle wedge peridotites underwent hydration by fluid derived from a dehydrated descending slab,and later interacted with seawater after emplacement at or near the seafloor.This study investigates how these two different rock-fluid interaction processes influenced trace element distribution in the southern Mariana forearc peridotites.We measured trace element concentrations of peridotites from the southern Mariana forearc.The southern Mariana forearc peridotites are characterized by a distinct seawater-like REE pattern with an obvious negative Ce anomaly,and La shows good correlation with other REEs (except Ce).In addition,there is a great enrichment of U,Pb,Sr and Li elements,which show a distinct positive anomaly relative to adjacent elements in the multi-element diagram.For the seawater-like REE pattern,we infer that REEs are mainly influenced by seawater during peridotite-seawater interactions after their emplacement at or near the seafloor,by serpentinization or by marine weathering.Furthermore,the anomalous behavior of Ce,compared with other rare earth elements in these samples,may indicate that they have undergone reactions involving Ce (IV) when the peridotites interacted with seawater.Positive U,Pb,Sr and Li anomalies are inferred to be related to seawater and/or fluids released during dehydration of the subducting slab.展开更多
Breeding strategies of Acrossocheilus fasciatus were studied from 592 specimens collected monthly during May 2007 and April 2008 in the Puxi Stream of the Huangshan Mountain. Sex ratio of the studied population was 0....Breeding strategies of Acrossocheilus fasciatus were studied from 592 specimens collected monthly during May 2007 and April 2008 in the Puxi Stream of the Huangshan Mountain. Sex ratio of the studied population was 0.90 ： 1 （ ♀ ： ♂ ）, not significantly different from 1 ： 1. Both sexes reached their first sexual maturity at age 2 （the second calendar year of their birth）. Fifty percent of females and males reached maturity at a total length of 69.75 mm and 69.36 mm respectively, and the minimum total length was 61.54 mm and 58.96 mm, respectively. Based on the monthly changes in gonado-somatic index and egg- development process, the breeding season of the population was from May to August, with one obvious interval （in June） occurring in the breeding activity for the females. The nonsynchronous development of oocytes observed in mature ovaries indicated that A. fasciatus is a batch spawner. Absolute fecundity of A. fasciatus ranged from 308 to 2002 eggs with a mean of 857 eggs, increased significantly with total length, and was significantly different among three age groups. Relative fecundity ranged from 38.63 to 71.70 egg/g with a mean of 53.29 egg/g, and was not significantly different among the three age groups. It was suggested that these reproductive characteristics were adaptive strategies for A. fasciatus to acclimatize to lotic water where environmental factors were unstable but predictable [ Current Zoology 55 （5） ： 350 - 356, 2009].展开更多
A phytochemical investigation on the aerial parts of Waltheria indica Linn.led to the isolation of 16 compounds,including five terpenoids(1–5),four coumarins(6–9),six flavonoids(10–15),and one phenylpropanoid glyco...A phytochemical investigation on the aerial parts of Waltheria indica Linn.led to the isolation of 16 compounds,including five terpenoids(1–5),four coumarins(6–9),six flavonoids(10–15),and one phenylpropanoid glycoside(16).The structures of these compounds were identified by spectroscopic data analysis and comparison with those reported in the literatures.Except for 12 and 15,all the compounds were isolated from W.indica for the first time.Moreover,coumarins were firstly reported to be obtained from this herb.The NO production inhibitory activities of all the isolated compounds in LPS-induced BV-2 cells were also presented.展开更多
基金the"95"great program of Chinese Academy of Sciences! (KY95 1-A1-3 0 1-0 2 )
文摘The cDNA encoding a variant of α bungarotoxin was cloned from the venom glands of Bungarus multicinctus by RT PCR.The deduced protein precursor contained a 21 amino acid signal peptide and a following 74 amino acid mature protein.The signal peptide is very similar to those of short chain neurotoxins,κ neurotoxins and cardiotoxins.The amino acid sequence of the mature protein is identical to α bungarotoxin (V31),a minor variant of α bungarotoxin identified by protein sequencing technique.Furthermore,the cDNA encoding the deletion precursor of α bungarotoxin was also cloned.By use of pMAL p2,the variant was overexpressed in E coli as a soluble fusion protein and purified by sepharose 6B amylose affinity chromatography,which was confirmed by western blotting with the antisera agai nst α bungarotoxin.The recombinant variant was achieved after digestion by factor X a.It displayed about 1/6 in vivo toxicity of natural α bungarotoxin.The successful cloning and functional expression of α bungarotoxin provided a basis for the future study of structure function of long neurotoxins.
文摘Aim To investigate the anticancer activity of two new cytotoxins from thevenom of Agkistrodon acutus. Methods The venom was isolated by FPLC column chromatography consistingof DEAE Sepharose FF and Source 30S. The cytotoxic activity on tumor cells was detected by MITmethod. Purity and molecular weight were determined by SDS-PAGE (silver staining). Their stabilitiesto temperature and pH were also detected. Results Two pure cytotoxins named ACTX-6 and ACTX-8 wereobtained. Their molecular weights are 98 kDa and 27 kDa, respectively. ACTX-6 consists of twosubunits bonded together by disulfide bonds. Conclusion ACTX-6 and ATCX-8 have highest inhibitoryactivity on lung cancer cell A549. ACTX-6 is stable to heat while ACTX-8 not. ACTX-6 is stablebetween pH 7-9 and ACTX-8 between pH 6 - 9.
文摘[Objective] This study aimed to obtain recombinant alpha-bungarotoxin （a-BG-0 gene fusion protein with biological activity and investiagte its fusion expression. [Method] The plasmid pGEX-a-BGT was transformed into E coil BL21 （DE3） and BL21 （DE3） plysS host bacteria to identify the optimal engineering strain. Fusion expression of the optimal engineering strain was induced, in order to optimize the induced expression conditions of the soluble fusion protein. [Result] JP-a-BGT was identified as the optimal engineering strain, which could express fusion protein after induced by IPTG. The optimal induced expression conditions of the soluble fusion protein were investigatect JP-a-BGT was incubated at 37 ℃ for 2.5 h and induced with 0.50 mmol4. IPTG for 4 h at 22 ℃, and the expression level of the soluble fusion protein reached 18.42%. [Conclusion] This study laid a solid foundation for the subsequent purification of fusion proteins and the separation and purification of a-BGT.
文摘Aim To study chemical constituents from Ampelopsis grossedentata. Methods Separation and purification were performed by using silica gel, polyamide, reverse-phase silica gel, Sephadex LH-20 column chromatographic techniques and silica gel PTLC. Structures were determined by means of physicochemical properties and spectral analysis. Results Four flavonoids were separated and identified from Ampelopsis grossedentata including dihydromyricetin (1), myricetin (2), myricitrin (3), and myricetin-3-O-β-D-galactopy...
文摘Total mRNA was extracted from a venom gland of snake Daboia russellii siamensis following the manufacturer＇s protocol of the PolyATtract System 1000 kit purchased from Promega Biotech. cDNAs encoding C-type lectins were amplified by RT-PCR and subcloned into a pMD18-T vector. Fourteen positive clones were selected for nucleotide sequencing and seven cDNAs encoding various snake venom C-type lectin-like protein precursors, designated as DRS-L1, DRS-L2, DRS-L3, DRS-L4 DRS-L5, DRS-L6 and DRS-L7, were obtained. Amino acid sequences of these proteins were deduced and each contains a carbohydrate recognition domain. Of all the deduced protein sequences, only DRS-L1 seemed to represent a closer sequence similarity to α subunits of other known snake venom C-type lectin-like proteins using the BLAST program. Homology comparison combined with analysis of cysteine position indicate that DRS-L1 and DRS-L2 are probably the light chain LC2 and LC1 of factor X activator from Daboia russellii siamensis venom, respectively. DRS-L3 and DRS-L4 might be the β subunits of higher molecular weight C-type lectin-like proteins while DRS-L5 and DRS-L6 might be β subunits of lower molecular weight C-type lectin-like proteins. DRS-L7 might be the β subunit of a platelet membrane glycoprotein Ib-binding protein similar to echicetin.
文摘A new angular furanocoumarin, 2′ acetylangelicin (8), along with oroselone (1), columbianadin (3), bergapten (4), O acetyl columbianetin (5), cniforin A (6), edultin (7) and β sitosterol (2), was isolated from the fruit of Cnidium monnieri. The structures were elucidated by spectral methods. Compound 1 was isolated from this plant for the first time.
文摘Objective： To investigate the effects of rAdinbitor on the adhesion and proliferation of human hepatoma cell strain 8MMC-7721. Methods： Cell adhesion assay was used to observe the effect of rAdinbitor on the adhesion of 8MMC-7721 cells to fibronectin （FN）. Crystal violet staining was performed to detect the influence of rAdinbitor on the adhesion of 8MMC- 7721 cells. MTT assay was employed to detect the inhibitory effects of different concentration of rAdinbitor on the proliferation of 8MMC-7721 cells. The morphologic changes of the control 8MMC-7721 cells and the apoptotic cells induced by 200μg/mL rAdinbitor for 36 h were observed under light microscope after HE staining. Flow cytometry analysis was applied to determine the apoptosis rate of 8MMC-7721 cells. Results： （1） FN promoted the adhesion of human hepatoma cell strain 8MMC-7721 in a dose-dependent manner. （2） rAdinbitor could dose-dependently inhibit the adhesion of SMMC-7721 cells to FN. The higher the concentration was, the stronger the inhibition was. There was significant difference among the groups （P 〈 0.05）. （3） rAdinbitor had a strong inhibition on the proliferation of 8MMC-7721 cells and showed a dose-dependent manner （P 〈 0.05）. After a 48 h exposure, the IC50 value of rAdinbitor was 177.83 μg/mL. （4） After exposure of 8MMC-7721 cells to 200μg/mL rAdinbitor for 36 h, the early morphologic changes appeared and the apoptosis rate was 20.68%, significantly higher than that of the control group （2.38%, P 〈 0.05）. Conclusion： rAdinbitor can dose-dependently inhibit the 8MMC-7721 cells adhesion to FN, and can inhibit the proliferation in dose-dependent manner and promote their apoptosis.
基金Supported by the Pilot Project of Knowledge Innovation Project,Chinese Academy of Sciences (Nos.KZCX2-YW-211, KZCX3-SW-223)the National Natural Science Foundation of China (No.40830849)the Special Foundation for the Eleventh Five-Year Plan of COMRA (No.DYXM-115-02-1-03)
文摘Peridotites from the southern Mariana forearc were sampled on the landward trench slope of the Izu-Bonin-Mariana (IBM) subduction zone by dredging.These mantle wedge peridotites underwent hydration by fluid derived from a dehydrated descending slab,and later interacted with seawater after emplacement at or near the seafloor.This study investigates how these two different rock-fluid interaction processes influenced trace element distribution in the southern Mariana forearc peridotites.We measured trace element concentrations of peridotites from the southern Mariana forearc.The southern Mariana forearc peridotites are characterized by a distinct seawater-like REE pattern with an obvious negative Ce anomaly,and La shows good correlation with other REEs (except Ce).In addition,there is a great enrichment of U,Pb,Sr and Li elements,which show a distinct positive anomaly relative to adjacent elements in the multi-element diagram.For the seawater-like REE pattern,we infer that REEs are mainly influenced by seawater during peridotite-seawater interactions after their emplacement at or near the seafloor,by serpentinization or by marine weathering.Furthermore,the anomalous behavior of Ce,compared with other rare earth elements in these samples,may indicate that they have undergone reactions involving Ce (IV) when the peridotites interacted with seawater.Positive U,Pb,Sr and Li anomalies are inferred to be related to seawater and/or fluids released during dehydration of the subducting slab.
基金by National Basic Research Program of China(2009CB119200)Anhui Provincial Natural Science Foundation(090413080)Natural Science Foundation of Anhui Education Bureau(KJ2009A110,KJ2008B211)
文摘Breeding strategies of Acrossocheilus fasciatus were studied from 592 specimens collected monthly during May 2007 and April 2008 in the Puxi Stream of the Huangshan Mountain. Sex ratio of the studied population was 0.90 ： 1 （ ♀ ： ♂ ）, not significantly different from 1 ： 1. Both sexes reached their first sexual maturity at age 2 （the second calendar year of their birth）. Fifty percent of females and males reached maturity at a total length of 69.75 mm and 69.36 mm respectively, and the minimum total length was 61.54 mm and 58.96 mm, respectively. Based on the monthly changes in gonado-somatic index and egg- development process, the breeding season of the population was from May to August, with one obvious interval （in June） occurring in the breeding activity for the females. The nonsynchronous development of oocytes observed in mature ovaries indicated that A. fasciatus is a batch spawner. Absolute fecundity of A. fasciatus ranged from 308 to 2002 eggs with a mean of 857 eggs, increased significantly with total length, and was significantly different among three age groups. Relative fecundity ranged from 38.63 to 71.70 egg/g with a mean of 53.29 egg/g, and was not significantly different among the three age groups. It was suggested that these reproductive characteristics were adaptive strategies for A. fasciatus to acclimatize to lotic water where environmental factors were unstable but predictable [ Current Zoology 55 （5） ： 350 - 356, 2009].
基金National Natural Science Foundation of China(Grant No.81473106 and 81773864)The Drug Innovation Major Project of China(Grant No.2018ZX09711001-008-003)
文摘A phytochemical investigation on the aerial parts of Waltheria indica Linn.led to the isolation of 16 compounds,including five terpenoids(1–5),four coumarins(6–9),six flavonoids(10–15),and one phenylpropanoid glycoside(16).The structures of these compounds were identified by spectroscopic data analysis and comparison with those reported in the literatures.Except for 12 and 15,all the compounds were isolated from W.indica for the first time.Moreover,coumarins were firstly reported to be obtained from this herb.The NO production inhibitory activities of all the isolated compounds in LPS-induced BV-2 cells were also presented.
