1.Progress in insertion sites for foreign sequence of foot and mouth disease virus.
Yan ZHANG ; Yonghao HU ; Fan YANG ; Haixue ZHENG
Chinese Journal of Biotechnology 2014;30(2):175-181
With the progess in studying gene structure and function of foot and mouth disease virus (FMDV), FMDV can express exogenous genes in different sites. Through transforming and modifying FMDV can achieve different application purposes such as improving virus titer, introducing tag, improving immune responses, and reducing pathogenicity. From the perspective of FMDV receiving inserted exogenous gene, this paper mainly describes the latest relevant developments of FMDV's expression to exogenous gene.
Foot-and-Mouth Disease Virus
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genetics
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Genetic Engineering
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Mutagenesis, Insertional
2.Flavanone O-glycosides from the rhizomes of Dryopteris sublaeta
Weisheng FENG ; Xinwei CAO ; Haixue KUANG ; Xiaoke ZHENG
Acta Pharmaceutica Sinica 2007;42(8):867-871
The aim of this study was to look for the chemical constituents from the rhizomes of Dryopteris sublaeta. The fresh plant was extracted twice with boiling water, the extract was concentrated to small volume under reduced pressure at 50 ℃. The concentrated material was partitioned with ether, ethyl acetate and n-butanol. The fraction of ethyl acetate was repeatedly chromatographied over silica gel and Sephadex LH-20 columns. Structures of pure compounds were established on the basis of their physiochemical and spectral data. Nine compounds were obtained and identified as sublaetentin A (1), sublaetentin B (2), sublaetentin C (3), sublaetentin D (4), matteuorienate A (5), matteuorienate C (6), arbutin (7), 3-methoxy-4-hydroxyphenyl-1-O-β-D-glucopyranoside (8) and 3,4-dimethoxyphenyl-1-O-β-D-glucopyranoside (9). Compounds 1-4 are new compounds, the others were isolated from this plant for the first time.
3.Chemical constituents from leaves of Celastrus gemmatus Loes.
Weisheng FENG ; Zhiyou HAO ; Xiaoke ZHENG ; Haixue KUANG
Acta Pharmaceutica Sinica 2007;42(6):625-630
To study the chemical constituents from the leaves of Celastrus gemmatus Loes., chromatographic methods were used to isolate and purify the chemical constituents, their structures were elucidated by the physiochemical characteristics and spectral data. Nine compounds were obtained and identified as (-)-massoniresinol 3a-O-β-D-glucopyranoside (1), ambrosidine (2), isolariciresinol 9-O-β-D-glucopyranoside (3), kaempferol 3-O-β-D-glucopyranoside(astragalin) (4), kaempferol 3-O-rutinoside (5), kaempferol 3-O-neohesperidoside (6), apigenin 7-O-β-D-glucuronide (7), apigenin 7-O-β-D-glucuronide methyl ester (8) and D-sorbitol (9). Compound 1 is a new compound, the others are isolated from this genus for the first time, and this is the first time to report lignan compounds from genus Celastrus.
4.A new stilbene glycoside from Dryopteris sublaeta
Weisheng FENG ; Xinwei CAO ; Haixue KUANG ; Xiaoke ZHENG
Acta Pharmaceutica Sinica 2005;40(12):1131-1134
Aim To study the chemical constituents of Dryopteris sublaeta Ching et Hsu. Methods Fresh plant of Dryopteris sublaeta Ching et Hsu was extracted twice with boiling water, concentrated to small volume under reduced pressure at 50 ℃. The concentrated material was partitioned with ether, ethyl acetate, and n-butanol. The fraction of ethyl acetate extract was chromatographed over macroporous adsorption resin (Diaion HP-20) eluted with a mixture of H2O and MeOH in increasing MeOH content.Their fractions from resin were repeatedly chromatographed over Toyopearl HW-40, Sephadex LH-20 and silica gel column chromatography. The compounds were identified on the basis of their physiochemical and spectral data. Results Four compounds were obtained and identified as 3,5-dihydroxy-stilbene-3-O-neohesperidoside ( 1 ), 3,5-dihydroxy-stilbene-3-O-β-D-glucoside ( 2 ), polydotin peceid (3) and 3,5,4'-trihydroxy-bibenzyl-3-O-β-D-glucoside (4). Conclusion Compound 1 is a new compound, the others were isolated from Dryopteris for the first time.
5.A new flavanone from Dryopteris sublaeta
Weisheng FENG ; Xinwei CAO ; Xiaoke ZHENG ; Haixue KUANG
Acta Pharmaceutica Sinica 2005;40(5):443-446
Aim To study the chemical constituents of Dryopteris sublaeta Ching et Hsu. Methods Fresh plant of Dryopteris sublaeta Ching et Hsu was extracted twice with boiling water, the extract was concentrated to small volume under reduced pressure at 50 ℃. The concentrated material was partitioned with ether, ethyl acetate, and n-butanol. The fraction of ether extract was chromatographed over silica gel column. The compounds were identified on the basis of their physiochemical and spectral data. Results Four compounds were obtained and identified as 2 (S)-5,7, 3'-trihydroxy-6,8-dimethy1-5'-methoxyflavanone ( 1 ), matteucinol ( 2 ), desmethoxymatteucinol ( 3 ) and 5,7,2'-trihydroxy-6,8-dimethy1-flavanone (4). Conclusion Compound 1 is a new one, the others were isolated from Dryopteris for the first time.
6.Experimental Study on Immunomodulatory Effect of Ethanol Sediments of the seeds of Descurainia sophia(L.) Webb.ex Prantl.
Xiaoke ZHENG ; Meng YANG ; Yiping BAI ; Yueting GUO ; Minghui ZHANG ; Haixue KUANG ; Weisheng FENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;17(3):507-513
This article was aimed to study the immunomodulatory effect of ethanol sediments of the seeds of Descurainia sophia(L.) Webb. ex Prantl. both in vitro and in vivo. The lymphocyte proliferation test in vitro was carried out to explore the effect of the ethanol sediments on the proliferation of T cell and B cell in the spleen of normal mice. And, the carbon clearance test, serum hemolysin test, and delayed-type hypersensitivity test were used to investigate the influence of fraction on non-specific immunity, humoral immunity and cellular immunity in the immunosuppressive mice induced by cyclophosphamide. Besides, the immunosuppressive model was used to evaluate the effect of fraction on immune organs and content of cellular factors in blood serum. The results showed that the ethanol sediments promoted Concanavalin A (Con A) induced T cell and Lipopolysaccharides (LPS) induced B cell (P < 0.01). It increased the carbon clearance index K, phagocytic index α, half value hemolysis (HC50), and swelling degree of auricula (P < 0.05 or P < 0.01). It reduced the body weight and atrophy of thymus and spleen index (P < 0.05 or P < 0.01). It increased the contents of IL-2, IFN-γ and IL-4 in serum in immunosuppressive mice (P < 0.05 or P < 0.01). It was concluded that ethanol sediments of the seeds of D. sophia(L.) Webb. ex Prantl. can boost the lymphocyte proliferation, protect the immune organs, and enhance the non-specific and specific immunity in immunosuppressive mice, which indicated that it had immune-promotion effect.
7.Inlfuence on Substance and Energy Metabolism of Diabetic Mouse Model of Chemical Split Fractions ofMori Cortex
Weisheng FENG ; Peipei YUAN ; Yan NIU ; Yingying KE ; Xiaolan WANG ; Zhiyi FENG ; Haixue KUANG ; Xiaoke ZHENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;17(3):464-470
This article was aimed to study the impact on substance and energy metabolism by chemical split fractions of Mori Cortex among hypoglycemic diabetic mouse model, in order to explain the new hypothesis of the science connotation in nature and flavor of traditional Chinese medicine (TCM). Male Kunming mice were intraperitoneally injected with a large dose of streptozotocin (STZ) (170 mg·kg-1) to establish type 1 diabetes mellitus mouse model. Medication was given consecutively for four weeks. The enzyme-linked immunesorbent assay (ELISA) was used to detect glucosekinase (GCK), glycogen phosphorylase (PYGL), pyruvate dehydrogenase (PDH), α-ketoglutarate dehydrogenase (α-KGDHC), phosphoglycerate kinase (PGK), acetyl coenzyme A (acetyl-CoA), adenylate kinase (ADK), fumarase (FUM), cytochrome C reductase (CCR), cytochrome C oxidase (COX) and other indicators. Enzymatic detection was used to determine the content of ATP coenzyme (ATPs), the content and ratio of NAD and NADH, the content of myocardial cell Na+-K+ ATP enzyme, as well as the content of ATP and ADP. The results showed that in the model group, the expression of PYGL was increased; and the expressions of GCK and PDH were decreased. It prompted that the source of glucose increased and the expelling of glucose decreased. The glucose level was increased. The COX expression was reduced and the respiratory chain was blocked. It regulated oxidative phosphorylation and the substrate phosphorylation level. It upregulated the expression of CCR, ATPs, NAD+, PGK, α-KGDHC and ADK. However, the expression of FUM was decreased. The activity of Na+-K+ ATPase was decreased significantly. At last, the metabolic disorders appeared. Mori Cortex aqueous extracts and the chemical split fractions significantly increased the GCK and PDH level in substance metabolism among diabetic mice. The levels of PYGL, α-KGDHC, PGK and acetyl-CoA were decreased (P < 0.05, or P <0.01). Meanwhile, it increased ATP and FUM, myocardial cell Na+-K+ ATP enzyme, and COX level in the energy metabolism (P < 0.05). It decreased the level of NAD+, CCR and ATPs (P < 0.05, or P <0.01). It was concluded that both the aqueous extracts and chemical split fractions of Mori Cortex can effectively improve the substance and energy metabolism disorders of diabetic mouse model. This effect may be related to the cold nature of Mori Cortex.
8.Expression of Major Antigen Domains of Gene of E2 CSFV and Analysis of its Immunological Activity
Hong TIAN ; Xiangtao LIU ; Jingyan WU ; Youjun SHANG ; Tao JIANG ; Haixue ZHENG ; Qingge XIE
Virologica Sinica 2008;23(4):247-254
E2 is an envelope glycoprotein of Classical swine fever virus (CSFV) and contains sequential neutralizing epitopes to induce virus-neutralizing antibodies and mount protective immunity in the natural host. In this study, four antigen domains (ABCD) of the E2 gene was cloned from CSFV Shimen strain into the retroviral vector pBABE puro and expressed in eukaryotic cell (PK15) by an retroviral gene expression system, and the activity of recombinant E2 protein to induce immune responses was evaluated in rabbits. The results indicated that recombinant E2 protein can be recognized by fluorescence antibodies of CSFV and CSFV positive serum (Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China) using Western blot, indirect immunofluorescence antibody test (IFAT) and ELISA, Furthermore, anti-CSFV specific antibodies and lymphocyte proliferation were elicited and increased by recombinant protein after vaccination. In the challenge test, all of rabbits vaccinated with recombinant protein and Chinese vaccine strain (C-strain) were fully protected from a rabbit spleen virus challenge. These results indicated that a retroviral-based epitope-vaccine carrying the major antigen domains of E2 is able to induce high level of epitope-specific antibodies and exhibits similar protective capability with that induced by the C-strain, and encourages further work towards the development of a vaccine against CSFV infection.
9.Chemical constituents from the leaves of Broussonetia papyrifera
Weisheng FENG ; Hongwei LI ; Xiaoke ZHENG ; Haixue KUANG ; Suiqing CHEN ; Yanzhi WANG ; Xinyu ZANG
Acta Pharmaceutica Sinica 2008;43(2):173-180
To separate and identify the chemical constituents from the leaves of Broussonetia papyrifera (Linn.) Vent, various columns including Diaion HP-20, Toyopearl HW-40C, Sephadex LH-20, silica gel were employed for the isolation and purification of compounds from the leaves of B.papyrifera. The structures of the compounds were elucidated by their physiochemical characteristics and spectral data. Nineteen compounds were isolated from the leaves of B.papyrifera and their structures were identified as apigenin (1), apigenin-7-O-β-D-glucopyranoside (2), chrysoerid-7-O-β-D-glucopyranoside (3), apigenin-7-O-β-D-glucopyranuronide (4), vitexin-7-O-β-D-glucopyranoside (5), luteolin (6), 5,7,4′-trihydroxyl-6-C-[a-L-rhamnopyranosyl(1→2)]-β-D-glucopyranosyl flavone (7), 5,7,4′-trihydroxyl-8-C-[a-L-rhamnopyranosyl(1→2)]-β-D-glucopyranosyl flavone (8), saponaretin (9), vitexin (10), benzyl benzoate-2,6-di-O-β-D-glucopyranoside (11), (2R,3R,5R,6S,9R)-3-hydroxy-5,6-epoxy-β-ionol-2-O-β-D-glucopyranoside (12), (2R,3R,5R,6S,9R)-3-hydroxyl-5,6-epoxy-acetyl-β-ionol-2-O-β-D-glucopyranoside (13), ficustriol (14), (6S,9S)-roseoside (15), 3β-hydroxy-5α,6α-epoxy-β-ionone-2α-O-β-D-glucopyranoside (16), icariside B1 (17), sammangaoside A (18), 3-hydroxy-5α,6α-epoxy-β-ionone (19). Compounds 11, 12 and 13 are new compounds, the others are isolated from this genus Broussonetia for the first time.
10.Expression and Immunological Analysis of Capsid Protein Precursor of Swine Vesicular Disease Virus HK/70
Hong TIAN ; Jingyan WU ; Youjun SHANG ; Shuanghui YING ; Haixue ZHENG ; Xiangtao LIU
Virologica Sinica 2010;25(3):206-212
VP1, a capsid protein of swine vesicular disease virus, was cloned from the SVDV HK/70 strain and inserted into retroviral vector pBABE puro, and expressed in PK15 cells by an retroviral expression system. The ability of the VP1 protein to induce an immune response was then evaluated in guinea pigs. Western blot and ELISA results indicated that the VP1 protein can be recognized by SVDV positive serum, Furthermore,anti-SVDV specific antibodies and lymphocyte proliferation were elicited and increased by VP1 protein after vaccination. These results encourage further work towards the development of a vaccine against SVDV infection.