1.Effects of intracere broventri cular injection of L-proline on learning and memory in mice
Danshen ZHANG ; Li ZHANG ; Shishan ZHANG
Chinese Pharmacological Bulletin 1987;0(03):-
In step-down tests in mice,the effects of L -proline 0.1 - 100?g ? mouse-1 icv on memory acquisition were not observed, but L -proline0. l~10?g ? mouse-1 icv could significantly improve the anisodine- induced impairments of memory acquisition. This effect of L-proline 1?g ? mouse-1 icv could not be antagonized with D-2-amino-5-phosphonovaleric acid(APS), the N-methyl-D-aspartate (NMDA) receptor antagonist. L-proline 1?g ? mouse-1 icv could also improve memory acquisition impairment induced by high GABA level in brain. This results suggested that L - proline may improve the memory as the modulator or possibly through GABA.
2.Study on chemical constituents from stem of Photinia parvifolia
Yuan LI ; Dongming ZHANG ; Shishan YU
Chinese Traditional and Herbal Drugs 1994;0(03):-
Object To study the chemical constituents from the stem of Photinia parvifolia (Pritz.) Schneid. Methods The compounds were isolated by silica gel column chromatography and their structures were elucidated by means of spectral analysis. Results Seven compounds were identified as lupeol (Ⅰ), ?-sitosterol (Ⅱ), betulinic acid (Ⅲ), pyracrenic acid (Ⅳ), epicatechin (Ⅴ), daucosterin (Ⅵ), pruasin (Ⅶ). Conclusion Compounds Ⅰ, Ⅲ-Ⅵ are isolated from the plants of Photinia Lindl. and compounds Ⅱ and Ⅶ are isolated from P. parvifolia for the first time. Compound Ⅳ shows anticancer effect.
3.Preparation and Quality Control of Weishu Emulsion
Xiaoming ZHANG ; Lin QIAN ; Shishan TANG
China Pharmacy 2001;0(09):-
OBJECTIVE:To prepare weishu emulsion and to establish its quality control method.METHODS:Weishu emulsion was prepared with sodium carboxymethylcellulose,etc.as the adjuvants and the content of dyclonine hydrochoride was determined by UV-spectrophotometry.RESULTS:The linear range of dyclonine hydrochloride was2.0~12.0?g/ml(r=0.9999)and the average recovery rate was99.38%,RSD=0.47%,(n=6).CONCLUSION:The emulsion is feasible in preparation method and controllable in quality.
4.Comparative Proteomic Analysis of the Promastigotes and Amastigotes of Leishmania donovani
Baoqian JING ; Shishan DENG ; Rengang ZHANG ; Jie ZHANG
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(02):-
Objective To explore the protein profile and identify developmentally regulated proteins of the promastigotes and axenic amastigotes with comparative proteomics technique.Methods The total proteins of promas-tigotes and axenic amastigotes of Leishmania donovani SC6 strain were separated by two-dimensional electrophoresis(2-DE) in a broad pH range(3-10) , and the gel was stained with Coomassie blue.The images were analyzed by PDQuest 1.0 software, and the major developmentally regulated proteins were identified by electrospray mass spectrometry.Results Approximately 700 protein spots were revealed in equivalent proteins of the promastigotes and axenic amastigotes separated by 2-DE, among which more than 90% protein spots showed equivalent quantity and distribution, with 6 proteins up-regulated and 3 proteins down-regulated in axenic amastigotes compared with promastigotes.Five of the 6 up-regulated proteins were with known function, respectively ascribed as Reiske iron-sulfur protein precursor, ?-tubulin, peroxidoxin 1, dihydrolipoamide acetyltransferase precursor, and mannose-1-phosphate guanyltransferase.Two of the 3 down-regulated proteins were identified as heat shock protein 70 and ?-tubulin.The functions of the developmentally regulated proteins were related to the carbohydrate/energy metabolism, stress response, or formation of cell membrane/cytoskeleton.Conclusion The findings demonstrate the differences in protein expression profiles between promastigotes and amastigotes.
5.Screening and identification of the mimic peptide of Mycobacterium tuberculosis antigen
Wenxia SUN ; Shishan YUAN ; Yunhong TAN ; Xiaoping ZHANG ; Yanyan YU
Chinese Journal of Microbiology and Immunology 2012;32(1):16-19
ObjectiveTo immunoscreen the mimic peptides of Mycobacterium tuberculosis antigen from phage displayed 12-mer peptide library.MethodsSpecific IgG was purified from sera of patients with TB and used as the target to immunoscreen a phage random peptide library of 12 amino acids.Positive clones which were obtained after three rounds of biopanning were detected by ELISA and sequenced.The diagnostic value of the high frequent positive clones were observed by ELISA.Results After 3 rounds of immunoscreening,the eluted phages were enriched effectively.Six kinds of animo acid sequence were obtained from twelve positive phage clones.Sensitivity of the two high frequent positive clones were 71.4% (A2)and 55.4% (A7) respectively.ConclusionThe antigen-mimic peptide was successfully screened from 12 random phage peptide library and the peptides can be recognized by tuberculosis patients' polyclonal antibodies.
6.Study on Diagnosis of Schistosomiasis by ELISA Using Periodate-treated Soluble Egg Antigen
Yuelong HUANG ; Xinyuan YI ; Xianfang ZENG ; Ran ZHANG ; Shishan YUAN
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(04):-
0. 05) and the specificity is higher than that of the SEA-ELISA (P
7.Pharmacodynamic Study of Yunkang Capsule
Zuoxiao LI ; Shunhan XIAO ; Sijin YANG ; Can TANG ; Shishan ZHANG
Traditional Chinese Drug Research & Clinical Pharmacology 2000;0(05):-
Objective To investigate the pharmacodynamic action of Yunkang Capsule(YKC).Methods The animals were divided into YKC groups(at high-,moderate-and low-dosage respectively),diphenidol control group,model control group and blank control group.The action of YKC on vertigo and thrombosis,hemorrheologic indexes,piamatral microcirculation and free radicals of the animal models were observed.Results YKC could significantly prolong the latency of vertigo,reduce wet weight of thrombus,decrease blood viscosity,plasma viscosity and hematocrit,promote the dilation of piamatral micro-vessels,increase the amount of interweave microdot,increase plasma SOD activity and decline plasma MDA content of animal models.Conclusion YKC has actions of counteracting vertigo and thrombosis,improving hemorrheology and microcirculation,and clearing away free radicals.
8.Blocking extracellular HMGB1 activity protects against doxorubicin induced cardiac injury in mice.
Yonggang MA ; Xiaowei ZHANG ; Huayan BAO ; Shishan YU ; Zhuowei HU ; Wei SUN
Acta Pharmaceutica Sinica 2012;47(11):1489-95
This study aims to investigate the preventive role and potential mechanisms of blocking extracellular HMGB1 function on doxorubicin induced cardiac injury. Mice were treated with HMGB1 blocker glycyrrhizin 1 h before and one time every day (intraperitoneal, 10 mg per mouse) after doxorubicin injection, and sacrificed on the day 14 after doxorubicin challenge. Cardiac function was evaluated by echocardiography and hemodynamic measurement. Myocardial inflammation and collagen deposition were analyzed by immunohistochemistry and picrosirius red staining. The interaction of HMGB1 and TLR2 was assessed by co-immunoprecipitation and confocal microscopy. The protein contents of HMGB1, MyD88, p65NF-kappaB and phospho-p65NF-kappaB were measured by Immunoblot. Compared with mice treated with saline, doxorubicin treatment led to an upregulation in HMGB1 expression. Blocking HMGB1 activity with glycyrrhizin protected mice against cardiac dysfunction, inflammatory response, and cardiac fibrosis induced by doxorubicin challenge. Glycyrrhizin inhibited the interaction of HMGB1 and TLR2, and blocked the downstream signaling of TLR2. In conclusion, blocking HMGB1 protected against doxorubicin induced cardiac injury by inhibiting TLR2 signaling pathway.
9.Expression and purification of SARS coronavirus membrane protein.
Wuxing, DAI ; Mingjun, LEI ; Shaoting, WU ; Zhihao, CHEN ; Liang, LIANG ; Huirong, PAN ; Li, QIN ; Shitong, GAO ; Shishan, YUAN ; Renli, ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(5):414-6
To construct a recombinant plasmid Pet23a-M, the gene encoding severe acute respiratory syndrome (SARS) coronavirus membrane protein was amplified by RT-PCR and cloned into the expression plasmid Pet23a. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis revealed that the cloned DNA sequence was the same as that reported. The recombinants were transformed into Escherichia coli (E. Coli) BL21 (DE3) and induced by Isopropyl-beta-D-thiogalactopyranoside (IPTG). The expression of 27 kD (1 kD=0.992 1 ku) protein was detected by SDS-PAGE and pured by metal chelated chromatography. Results of Western-blot showed that this expressed protein could react with antibodies in sera of SARS patients during convalescence. This provided the basis for the further study on SARS virus vaccine and diagnostic agents.
Cloning, Molecular
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Escherichia coli/genetics
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Escherichia coli/metabolism
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Membrane Proteins/*biosynthesis
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Membrane Proteins/genetics
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Membrane Proteins/isolation & purification
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Plasmids/biosynthesis
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Plasmids/genetics
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Recombinant Proteins/biosynthesis
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Recombinant Proteins/genetics
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Recombinant Proteins/isolation & purification
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Reverse Transcriptase Polymerase Chain Reaction
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SARS Virus/chemistry
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SARS Virus/*genetics
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Viral Vaccines/biosynthesis
10.Construction and application of chimeric infectious clones of porcine reproductive and respiratory syndrome virus.
Xiangjian LI ; Jianwu ZHANG ; Jian LÜ ; Dandan YU ; Huochun YAO ; Shishan YUAN
Chinese Journal of Biotechnology 2008;24(9):1573-1581
In recent years, mass outbreaks of highly pathogenic (HP) porcine reproductive and respiratory syndrome virus (PRRSV) have spread all over the Chinese swine industry. Based on the first infectious cDNA clone of HP PRRSV strain pJX143 and that of an attenuated PRRSV, pAPRRS, constructed in our group, we constructed several chimeric clones with various substitutions of structural protein genes (ORF4-7) and 3' UTR between attenuated pAPRRS and virulent pJX143.Upon transfection of MA-104 cultured cells, all chimeric constructs pSX12, p5NX12, and p56N12 were rescued. The rescued viruses maintained the similar virological properties, based on the results of the growth curve of the rescued viruses. To test if the chimeric viruses can be used as a vaccine candidate, vSX12 and v56N12 vaccinated pigs were challenged with the HP PRRSV JX143 strain. As a result, the vSX12 vaccinated pigs were all seroconverted by 14-day-post vaccination, while v56N12 vaccinated pigs showed poor antibody response. Upon challenge, the vSX12-vaccinated group showed no signs of clinical PRRS syndrome, and virema period was shorten to 6 days post-challenge. Our results demonstrated that 1) vSX12 chimeric virus is a good vaccine candidate; 2) the virulence determinants of HP PRRSV probably located in coding regions other than ORF3-7 and 3' UTR, as our chimeric viruses were proved to be attenuated.
Animals
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Cloning, Molecular
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Open Reading Frames
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Porcine Reproductive and Respiratory Syndrome
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virology
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Porcine respiratory and reproductive syndrome virus
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genetics
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immunology
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Recombinant Fusion Proteins
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genetics
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immunology
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Recombination, Genetic
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genetics
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Swine
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Vaccines, Attenuated
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immunology
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Viral Envelope Proteins
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Viral Proteins
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biosynthesis
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genetics
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Viral Vaccines
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immunology