2.Effect of Hedysarum Polybotys Saccharide on Insulin Resisance of Experimental NIDDM Rats
Zhi-Sheng JIN ; Juan-E LI ; Dong-Peng ZHANG ;
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(05):-
Objective To investigate the effect of Hedysarum Polybotys saccharide(HPS)on insulin resistance rats.Methods The model of type 2 diabetic-insulin resistance rat was successfully made by STZ and high caloric diet.Then the rats were intervene by Metformin(MT)and HPS.Results Compared with the model group,the level of blood glucose,INS and CP were lowered,ISI were improved and FFA were restrained by HPS.Conc(?)sion HPS can reduce the insulin resistance in multiple ways.
3.Recognition of vitiligo associated protein VTT-1 gene and its sequence analysis
Yong-Wei LI ; Di-Min ZHANG ; Xiao-Dong WEI ; Yi-Jin ZHANG ; Ai-E XU ;
Chinese Journal of Dermatology 1995;0(04):-
Objective To investigate the expression of VIT-1 gene in melanocytes of patients with vitiligo, and to analyze the difference of its sequence. Methods The skin from the foreskins of healthy men by circumcision and from the non-lesional area on the buttocks of 5 patients were digested by dispase, then the epidermis and dermis were separated, and the melanocytes were isolated. Then we cultured the melanocytes from the controls in TICVA medium and those from the patients in TICVA medium supplemented with basic fibroblast growth factor (bFGF) and endothelin-1 ( ET-1). The expression of VIT-1 gene was measured by RT-PCR, the full-length cDNA of VIT-1 ORF was cloned and sequenced, and sequence difference was analyzed by CLUSTAL W ( 1.83 ) software. Results The expression levels of VIT-1 gene were significantly lower in melanocytes from the patients than in those from the controls. An 81 bp-intron was found in the VIT-1 ORF. VIT-1 was a fragment of FBXO11, located at its 3' end. Conclusion VIT-1 gene is not a new gene, but a fragment of FBXO11, and a member of F-box protein family.
4.Preparation of corn polysaccharide-Fe(Ⅲ) complex and assay of Fe(Ⅲ)
xiao-lei, DENG ; jian-hua, ZHANG ; jin-e, ZHOU ; ze-nai, CHEN ; yang, LU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(11):-
11.0 for the pH of reaction solution,65℃~75℃for the reaction temperature,and 1:1.7 for the mass ratio of corn(dry weight)to FeCl_3?6H_2O.The corn polysaccharide-Fe(Ⅲ)complex synthesized with the optimized process was stable,with good solubility in water.The assay of Fe(Ⅲ)was 39.86%,40.20%and 40.17%,respectively for three batches of products.The RSD was
5.Expression of retinoic acid-related orphan receptors and interleukin-17 in mice with vaginal candidiasis
Dan LUO ; Jin′e ZHANG ; Rongyi CHEN ; Yanping YANG ; Ying ZHOU ; Yiming FAN ;
Chinese Journal of Dermatology 2017;50(1):33-38
Objective To investigate the role of T helper 17 cells/interleukin?17(Th17/IL?17) axis in the occurrence of vaginal candidiasis in mice. Methods A total of 120 female BALB/c mice were randomly and equally divided into Ei, En, Ci and Cn groups. Three days before vaginal inoculation, estrogen (Ei and En)groups and control(Ci and Cn)groups received subcutaneous injection of 0.05 mg estradiol and 0.1 ml sterilized soybean oil at the hind legs, respectively, and then the hormone treatment continued every other day until the end of experiment. Infected(Ei and Ci)groups and noninfected(En and Cn) groups were inoculated intravaginally with 10μl(5 × 104 conidia)of Candida albicans suspension and 10μl of sterilized phosphate?buffered saline, respectively. Ten mice were randomly selected from each group and sacrificed on day 3, 7 and 14 after inoculation. The intact vagina tissues were resected and then frozen in liquid nitrogen or embedded in paraffin. Real?time fluorescence?based quantitative PCR(qRT?PCR)and immunofluorescent staining were performed to measure mRNA expression and immunofluorescence intensities of retinoic acid?related orphan receptorγt(RORγt), RORα and IL?17, respectively. Western blot analysis was conducted to determine protein expression of RORγt and IL?17. Results Laser scanning confocal microscopy showed that RORγt, RORα and IL?17 immunofluorescence was mainly located at inflammatory cells of the lamina propria and blood vessels in En and Cn groups, at mucosal epithelium, adherent hyphae, and inflammatory cells of the lamina propria and blood vessels in Ci group, and at mucosal epithelium, vaginal canal and endocytosed hyphae, and inflammatory cells of the lamina propria and blood vessels in Ei group. qRT?PCR and immunofluorescent staining uncovered that mRNA expression and immunofluorescence intensities of RORγt, RORα and IL?17 were significantly higher in En, Ci and Ei groups than in Cn group at the same time points(all P<0.05), as well as in the Ei group than in En and Ci groups(both P<0.05), and were increased gradually over time in En, Ci and Ei groups, but not in the Cn group. Additionally, mRNA expression and immunofluorescence intensities of RORγt and RORαand IL?17 generally peaked on day 14 after inoculation, while the immunofluorescence intensity of IL?17 peaked on day 7 (P < 0.05). Western blot analysis revealed that protein expression of RORγt and IL?17 was significantly higher in the infected(Ei and Ci)groups than in the noninfected(En and Cn)groups at the same time points(RORγt:F=45.685, P<0.001;IL?17:F=29.655, P<0.01), and was highest in the Ei group(P<0.05);however, no significant differences were observed between Cn and En groups(both P>0.05). Moreover, RORγt and IL?17 protein expression in Ci and Ei groups was obviously up?regulated on day 7 after inoculation (RORγt: F = 13.137, P < 0.001; IL?17: F = 11.182, P < 0.001), but was not increased further on day 14. Conclusion Vaginal candida infection can up?regulate the expression of RORγt, RORα and IL?17, suggesting that Th17/IL?17 axis may be involved in the occurrence of vaginal candidiasis in BALB/c mice.
6.An improved association-mining research for exploring Chinese herbal property theory: based on data of the Shennong's Classic of Materia Medica.
Rui JIN ; E-mail: ZHANGBING6@263.NET. ; Zhi-jian LIN ; Chun-miao XUE ; Bing ZHANG
Journal of Integrative Medicine 2013;11(5):352-365
Knowledge Discovery in Databases is gaining attention and raising new hopes for traditional Chinese medicine (TCM) researchers. It is a useful tool in understanding and deciphering TCM theories. Aiming for a better understanding of Chinese herbal property theory (CHPT), this paper performed an improved association rule learning to analyze semistructured text in the book entitled Shennong's Classic of Materia Medica. The text was firstly annotated and transformed to well-structured multidimensional data. Subsequently, an Apriori algorithm was employed for producing association rules after the sensitivity analysis of parameters. From the confirmed 120 resulting rules that described the intrinsic relationships between herbal property (qi, flavor and their combinations) and herbal efficacy, two novel fundamental principles underlying CHPT were acquired and further elucidated: (1) the many-to-one mapping of herbal efficacy to herbal property; (2) the nonrandom overlap between the related efficacy of qi and flavor. This work provided an innovative knowledge about CHPT, which would be helpful for its modern research.
Data Mining
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Drugs, Chinese Herbal
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Humans
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Materia Medica
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Medicine, Chinese Traditional
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Qi
8.Cloning and expressing of cellulase gene (cbh2) from thermophilic fungi Chaetomium thermophilum CT2.
Shou-An LIU ; Duo-Chuan LI ; Shi-Jin E ; Yan ZHANG
Chinese Journal of Biotechnology 2005;21(6):892-899
Chaetomium thermophilum CT2 can produce extracellular cellulase with industrial value. We designed two degenerate primers to amplify catalytic domain sequence of cellobiohydrolase II ( CBH II). Full length of cDNA was obtained by rapid amplification of cDNA ends technologies. DNA sequencing revealed that cbh2 has an open reading frame of 1428bp, which encodes a putative polypeptide of 476 amino acids. The deduced amino acid sequence shows that the predicted molecular mass is 53 kD and the cbh2 consists of a fungal-type carbohydrate binding domain (CBD) separated from a catalytic domain by a linker region rich in proline/serine/threonine. PCR product consisting of the entire CBH II coding region without its signal sequences was cloned into the yeast secretive plasmid pPIC9K, which was then transformed into Pichia pastoris GS115. Highly efficient production of the cellobiohydrolase II was achieved in P. pastoris under the control of the AOX1 promoter, and the expressing level was 1.2 mg/mL by small-scale culturing. The recombinant cellobiohydrolase II was purified by using ammonium sulfate fraction, DEAE-Sepharose Fast flow chromatography. A molecular mass of the purified enzyme is 67 kD determined by SDS-PAGE and this is similar to the native cellobiohydrolase II purified from C. thermophilum CT2. The recombinant enzyme exhibited optimum catalytic activity at pH 4.0 and 50 degrees C respectively. It was thermostable at 50 degrees C and retained 50% of its original activity after 30 min at 70 d degrees C . The high level of fully active recombinant cellobiohydrolase II got from P. pastoris makes this expression system attractive for fermentor and industrial applications.
Amino Acid Sequence
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Base Sequence
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Cellulose 1,4-beta-Cellobiosidase
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biosynthesis
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genetics
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Chaetomium
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enzymology
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genetics
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Cloning, Molecular
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DNA, Complementary
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genetics
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Fungal Proteins
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biosynthesis
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genetics
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Molecular Sequence Data
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Open Reading Frames
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genetics
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Pichia
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genetics
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metabolism
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Recombinant Proteins
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biosynthesis
;
genetics
10.Expression of p57 and homeobox A10 during decidualization of endometrial stromal cell in vitro..
Kun QIAN ; E-mail: ZHU_GUIJIN@SINA.COM. ; Hong CHEN ; Han-Wang ZHANG ; Yu-Feng LI ; Lei JIN ; Gui-Jin ZHU
Acta Physiologica Sinica 2005;57(4):498-504
In order to elucidate the function of homeobox A10 gene (HOXA10) and p57 during decidualization our present study was designed to observe the change of HOXA10 and p57 expression and subcellular localization of HOXA10 in the process of endometrial stromal cell (ESC) differentiation in vitro. Decidualization was induced by 0.5 mmol/L 8-Bromo-cAMP (8-Br-cAMP) together with 1x10(-6) mol/L medroxyprogesterone acetate (MPA). Expression of p57 and HOXA10 was detected by RT-PCR and Western blot after 1-day, 2-day, and 4-day treatment (D1, D2, D4). ESCs cultured in 2%FBS for 1 and 4 d were used as control (C1, C4). The location of HOXA10 was detected by indirect immunofluorescence and HOXA10-GFP transfection. The results are as follows: (1) The expression of HOXA10 decreased progressively during the course of decidualization, and showed significant difference compared to control group C4 after 2-day treatment (D2). (2) On the contrary, the expression of p57 increased progressively and also showed significant difference compared to the control group C4 after 2-day treatment (D2). (3) There was no significant change of HOXA10 and p57 expression after culturing ESCs in 2%FBS for 4 d (C1, C4). (4) HOXA10 located in the nucleus throughout the course. Cytoplasm and nucleus shuttle was not detected in the experiment. Our results suggest that the down-regulation of HOXA10 may contribute to the increase of p57 and that the up-regulation of p57 likely plays an important role in ESC differentiation. Progesterone receptor (PR) pathway may participate in promoting ESCs to exit cell cycle and enter differentiation.