2.RESEARCH ON THE TESTING OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS BY NESTRT-PCR
Yun ZHAO ; Guang-Chuan ZHANG ; Yu-Ming QIN ; Yi-Bao NING ; Qin WANG ;
Microbiology 1992;0(06):-
Three primer were designed based on the consevered area of the genetic of the ATCC VR-2332 strain and LV strain. And the nest RT-PCR of testing porcine reproductive and respiratory syndrome virus were developed. The nest RT-PCR against ATCC VR-2332 strain, LV strain and B13 strain were done by this method.The DNA fragment were obtained specially from the three strains isolated from different region. The size were 430bp (430bp) , 410bp (413bp) and 410 bp (413bp) separately. But the DNA fragment were not obtained from HCV, PPV and PRV. Its sensitivity was 10-2 TCID50. It's sensitivity increased 10000 times than one step RT-PCR. It should make the method of testing porcine reproductive and respiratory syndrome virus more sensitive, fast and accurate.
3.Differentiation Between Vaccine Strain and Field Isolates of Classical Swine Fever Virus Using Polymerase Chain Reaction and Restriction Test
Yun ZHAO ; Yu-Ming QIN ; Guang-Chuan ZHANG ; Qi-Zu ZHAO ; Yi-Bao NIN ;
Microbiology 1992;0(03):-
A nest RT-PCR/restriction test has been developed in order to distinguish the lapinised vaccine strain from field isolates of classical swine fever virus. The restriction enzyme cut sites mapping of the major coding sequence of E2 gene lapinised vaccine strain and ShiMen strain of classical swine fever virus have been compared. Ten and sixteen unique restriction markers have been found in the lapinised vaccine strain and ShiMen strain. The restriction enzyme cut sites mapping of the twenty six unique restriction marker in the major coding sequence of E2 gene of 17 classical swine fever field isolates have been analyzed. Only 3 sites (HgaI、Hin8I及Hsp92I) are present in the lapinised vaccine strain sequence. Two pans of nested primers and a criteria of analysis have been designed for HgaI restriction marker site. The tests have been conducted first on the lapinised vaccine strain and ShiMen strain of classical swine fever virus resulting in predicted restrection patterns. Finally, the tests have been applied to 5 field isolates of different gene group analyzed by phylogenetic study. The result showed that only HCLV strain gene can be cut to 2 fragment by Hgal , and ShiMen strain and 5 field isolates cant be cut At the same time the sensitivity and specificity of nest RT-PCR have been tested. The sensitivity is 0. 2MLD. The specific fragment of BDV and BVDV were not obtained by the nest RT-PCR. These results showed that the development of the nest RT-PCR/restriction tests is very important for the control and perish of classical swine fever in china.
4.Synthesis and activity of ACE inhibitory peptides.
Jin REN ; Gang CAO ; Rui-Jie ZHANG ; Da-Wei LI ; Ting-Ting WEI ; Chuan-Guang QIN
Acta Pharmaceutica Sinica 2011;46(1):58-63
To find anti-hypertensive lead drug, angiotensin converting enzyme (ACE) inhibitory peptides were synthesized and their effects on inhibiting ACE activity were investigated. ACE inhibitory peptides were synthesized via Fmoc solid-phase synthesis, isolated and purified through reversed phase high-performance liquid chromatography (RP-HPLC), and identified by mass spectrometry. A RP-HPLC analysis method was used to test ACE inhibitory activity in vitro of these ACE inhibitory peptides. Six octapeptides were successfully synthesized, and the analytical results of mass spectrum were consistent with their theoretically calculated data. Among these synthetic octapeptides, the anti-SARS (severe acute respiratory syndromes) octapeptide had the most obvious ACE inhibitory activity with an IC50 value of 3.4 x 10(-5) mol x L(-1). So octapeptide AVLQSGFR-OH (anti-SARS peptide) was found to be the strongest candidate for potential development as an anti-hypertensive drug and had the implication of further study.
Angiotensin-Converting Enzyme Inhibitors
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chemical synthesis
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chemistry
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pharmacology
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Antihypertensive Agents
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chemical synthesis
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chemistry
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pharmacology
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Chromatography, High Pressure Liquid
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methods
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Mass Spectrometry
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Molecular Structure
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Oligopeptides
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chemical synthesis
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chemistry
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pharmacology
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Peptidyl-Dipeptidase A
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drug effects
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Solid-Phase Synthesis Techniques
5.Development of cell-penetrating peptides as vectors for drug delivery.
Jin REN ; Chuan-Guang QIN ; Chun-Lan XU ; Qiu-Yu WANG ; Xiao-Jia ZUO
Acta Pharmaceutica Sinica 2010;45(1):17-25
Biomacromolecules play an important role in the treatment of many diseases, but as a result of cell membrane serving as the natural barriers, only the small molecular compounds whose molecular weights are smaller than 600 Da can get through cell membrane and enter the cell. In recent years, some short peptides (the length less than 30 amino acids) are found to have the cell-penetrating function, called cell-penetrating peptides (CPPs). They are able to effectively translocate segments of protein, polypeptides, nucleic acid into the cells of many mammal animals with many methods. They have high transduction efficiency and will not lead to cell damage. So, the discovery of CPPs has a very good applicable prospect in such research fields as cell-biology, gene-therapy, drug transduction in vivo, evaluation of clinical medicine and medical immunology. This paper reviews the types and characteristics of CPPs, internalization mechanisms, applications, and their existing problems.
Absorption
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drug effects
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Amino Acid Sequence
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Animals
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Cell Membrane Permeability
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Cell-Penetrating Peptides
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classification
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pharmacology
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physiology
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Drug Carriers
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Endocytosis
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physiology
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Humans
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Protein Transport
6.Study on relationship between length of homologous sequences and chromosomic recombination rate in Saccharopolyspora erythraea.
Bu-Chang ZHANG ; Zhi-Hu ZHAO ; Yi-Guang WANG ; Xiu-Qin YU ; Chuan-Xuan LIU ; Qing-Jun MA
Chinese Journal of Biotechnology 2003;19(1):13-18
In order to study the relationship between lengths of homologous fragments and chromsomic recombination rate in Saccharopolyspora erythraea, three homologous sequences, with mutant loci and different flanking sequences, (26bp + 27bp), (500bp + 576bp) and (1908bp + 1749bp), were synthesized by chemical reaction or PCR amplification, and cloned into pWHM3 to construct homologous recombination plasmids, pWHM1113, pWHM1116 and pWHM1119. When the plasmids were transformed into protoplast of Saccharopolyspora erythraea A226 under PEG mediated, on an average 30, 69 and 170 transformants grew on each plate for the three plasmids respectively, but chromosomic integration frequency were 0, 2% and 19% among corresponding transformants. Both pWHM1116 and pWHM1119 could take double crossover recombination, and exchange the mutant loci in the chromosome. It was concluded that when the flanking sequences were equal or more than (500bp + 576bp), they could take effective single and double recombination with Saccharopolyspora erythraea chromosome.
Chromosomes, Bacterial
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genetics
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Polymerase Chain Reaction
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Recombination, Genetic
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genetics
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Saccharopolyspora
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genetics
7.The observation on the relationship between iron deficiency and altitude hypoxia
zhen-ting, QIN ; li-yang, SHEN ; hong-cai, MIAO ; ji-chuan, LIU ; li-ming, LIN ; er-dao, GE ; Gage DUSEK ; ci, WEI ; guang-fu, YUAN
Journal of Applied Clinical Pediatrics 1986;0(01):-
Background Since the measurement method establishment of serum ferritin abroad in early period of theseventies, the iron deficiency had been divided into two types: the non-anemia and anemia types. In orderto go step further studies, we must ertablish the bemoglobin targets of the two types. Methods One hurdred and fifty-two children in experimontal group, from 6 to 7 years old, and allcome from Qinghai province. There are 29 children in Xining city, 24 in Guide, 26 in Gongbe, 40 in Gui-nan and 33 in Maduo countics. There are 36 health children aged from 6 to 7 years old in the controlgroup, and all comes from Beijing. The Hb, RBC, HCT, HCTW and FEP wcre determined. Results The three targets correlating with Hb (Hb, MCH and MCHC); correlating with RBC (RBC,HCT and MCV); the two targets correlating with RBC_weight (HCTW and CMCW) and correlating withFEP of RBC(FEP and MCEP) have very significant difference between experimental group and control group. Conclusion The determination values of the 10 targets are not same in children in different districts,and the values of all the target: are increased on different degree along with the increase in altitude of ele-vation. There is very important significance on the studies of iron deficiency and altitude hypoxia to establish the normal values of the 10 targets.
8.Experiment on the sexual virility of immature mice immunized with LHRH fusion protein vaccine.
Qin-Qin PAN ; Yu-Gui CUI ; Guang-Ming GONG ; Chuan-Yi JI ; Yong PAN ; Xing-Hai WANG
National Journal of Andrology 2005;11(8):566-573
OBJECTIVETo observe the sexual virility of immature immature male mice were divided into a pre-ablactation group (n 10). The first two groups were immunized with the LHRH fusion proportion of pregnant female mice, the morphological and histological examined to conform the emasculating effect of the vaccine. When ted with testosterone (1.0 ml each) , the post-ablactation ones were rameters.
RESULTSThe sexual virility of the immature mice immunized in 3 -4 months.
CONCLUSIONThe LHRH fusion protein vaccine mice after ablactation, and the sexual virility can recover in the pre-ablactation decrease.
Animals ; Antibodies ; metabolism ; Female ; Gonadotropin-Releasing Hormone ; immunology ; Male ; Mice ; Mice, Inbred ICR ; Pregnancy ; Recombinant Fusion Proteins ; immunology ; Sexual Development ; immunology ; Sexual Maturation ; immunology ; Sperm Motility ; Testosterone ; analysis ; Vaccines, Synthetic ; immunology
9.Detection and evaluation of the mutations of embB gene in ethambutol-susceptible and resistant Mycobacterium tuberculosis isolates from China.
Xue-Qiong WU ; Jian-Qin LIANG ; Jun-Xian ZHANG ; Yang LU ; Hong-Min LI ; Guang-Yu ZHANG ; Guo-Rui YAN ; Bei-Chuan DING
Chinese Medical Journal 2005;118(20):1739-1741