1.High-sensitive Detection Method for Siderophores from Pseudomonas
Shao-Xing CHEN ; Xiang ZHAO ; Ping SHEN ; Zhi-Xiong XIE ;
Microbiology 1992;0(03):-
CAS (Chrome azurol S) assay was a universal method for detecting the bacterial siderophores, and Sugar-Asn liquid medium has been applied to the studies on siderophores from Pieudomonas. In this paper, Asp has been substituted for Asn, and MSA-CAS agar plate was developed by integrating the MSA (sugar-Asp) medium and CAS bright blue dye, which has been used in the universal CAS assay. On the aspect of siderophores detection , 8 strains of 7 species from Pseudomonas had been screened on MSA-CAS agar plates and universal CAS assay respectively. The results showed that MSA-CAS agar was higher-sensitive and lower basic fluorescent than universal CAS assay.
2.Improvement and Analysis in Over- siderophores Production Bacteria Filtrating and Detecting
Xiang ZHAO ; Zhi-Xiong XIE ; Shao-Xing CHEN ; Ping SHEN ;
Microbiology 1992;0(06):-
A novel stable blue agar plate which is convenient to preparing and more effective than the universal chrome azurol sulfonate (CAS) assay established by Schwyn and Neilands was designed by replacing MM9 growth medium and pipes with certain concentrate of phosphorus buffer solution which pH could be stabled at 6.8. It is more suitable for screening over- siderophores production bacteria. Since OD_ 630 of the sample is usually out of the range of spectrophotometer with CAS assay solution when quantifying the siderophores and the outcome is not steady,the measuring wavelength had been changed to 680 nm corresponding to the middle of max absorbance and the correlation between siderophores concentrations and OD was unchanged. But the detecting sensitivity is elevated by enlarged the absorbance differences among samples with different productivity of siderophores at 680 nm .
3.Eukaryotic Expression of ?ig-h_3 Gene and Its Effects on Secretion of MMPs in the Human Hepatoma 7721 Cells
Juan TANG ; Jian-Li JIANG ; Hong-Wei ZHOU ; Hua XIONG ; Xiang-Min YANG ; Zhi-Nan CHEN ;
China Biotechnology 2006;0(09):-
?ig-h_3 was first identified as a transforming growth factor-beta1-inducible gene in human lung adenocarcinoma cell line. It encodes for a secreted extracellular matrix (ECM) protein, which is thought to act on cell attachment and ECM composition. Previous study showed that ?ig-h_3 were highly expressed in human hepatoma cell lines and lowly expressed in human normal hepatic cells. The present study aimed to transfect ?ig-h_3 into 7721 cells to investigate its effect on secretion of MMPs in the transfected human hepatoma cells. Full-length ?ig-h_3 gene,cloned by reverse transcription polymerase chain reaction (RT-PCR) was inserted into the eukaryotic expression vector pEGFP-C_2. The recombinant plasmid was transfected into 7721 cells with Lipofectamine2000 and Gelatin-Zymography were adopted to detect the production of MMPs in the transfected cells. Results showed that ?ig-h_3/pEGFP-C_2 recombinant expression plasmid was successfully constructed and achieved high transfection efficiency. MMPs expression of the transfected cells was promoted significantly. These results suggest that overexpression of ?ig-h_3 promoted the production of MMPs, indicating that ?ig-h_3 may play roles in the invasive and metastatic processes of hepatoma.
4.HLA-DQA1 genotyping by PCR-SSP technique in Jing nationality of Central Vietnam
Ting-Ping CHEN ; Lin-Lin WANG ; Wei-Xiong LIN ; Jian-Feng CHEN ; Xiang-Zhi XIE ;
Chinese Journal of Immunology 1985;0(03):-
Objective:To investigate the polymorphism of HLA-DQA1 genes in Jing nationality of Central Vietnam.Methods:Applied PCR-SSP tecnique to determine the polymorphism of the HLA-DQA1 alleles of 105 healthy children and youth,unrelated individuals in Central of Vietnam.Results:10 HLA-DQA1 alleles were detected of which DQA1*0104 were the most common allele with frequency of 21.3% and lowest frequency is DQA1*0601.Conclusion:The results indicate that HLA-DQA1 alleles polymorphism of Jing nationality in Central Vietnam is different from the other Chinese. [
5.Enhancement of artemisinin biosynthesis in transgenic Artemisia annua L. by overexpressed HDR and ADS genes.
Ya-Xiong WANG ; Shi-Ping LONG ; Li-Xia ZENG ; Li-En XIANG ; Zhi LIN ; Min CHEN ; Zhi-Hua LIAO
Acta Pharmaceutica Sinica 2014;49(9):1346-1352
Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.
Artemisia annua
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genetics
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metabolism
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Artemisinins
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metabolism
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Biosynthetic Pathways
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Drugs, Chinese Herbal
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Mixed Function Oxygenases
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genetics
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Oxidoreductases
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genetics
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Plant Proteins
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genetics
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Plants, Genetically Modified
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genetics
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metabolism
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Plants, Medicinal
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genetics
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metabolism
6.Investigation into the serum uric acid level of the residents in Henglan town of Zhongshan city.
Chun-lin XIONG ; Wen LI ; Jun CHEN ; Zhi-liang WU ; Qing-xiang HOU
Journal of Southern Medical University 2009;29(4):798-800
OBJECTIVETo investigate the relationship between cardiac Cerebrovascular disease and serum uric acid(SUA) in the coastal inhabitant, and try to provide base for prevention of the local metabolic disease and cardiac Cerebrovascular disease.
METHODWe got 3111 local people who had participated in the annual physical examination in the perch hospital for the research on the level of SUA and the relative risk factor. According to SUA level we divided the cases into two groups. One is hyperuricemia group and the other is control group.
RESULTS(1)The average SUA level was (380.2-/+62.58) micromol/L in the males, while (290.82-/+60.32) micromlo/L in the female. The sick rate of hyperuricaemia rate, was 21.8% for male, and the 17.6% of female. This disease affected more men than women. It has significant difference (P<0.01); (2)The SUA level of the showed a positive correlation with the indexes of the total cholesterol, the triglyceride, the blood pressure and the body weight index. The SUA level in overweight/obesity people was obviously higher than that of the normal weight people.
CONCLUSIONS(1)The sick rate of hyperuricaemia is high in Zhongshan coastal area, which due to many related factors. As far as, few people know it, so we should adopt the synthesis measure to prevent and control it to reduce the sick rate; (2)the rise of The SUA level possibly becomes one of the independent dangerous and predictive factor for the heart cerebral von disease.
Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Case-Control Studies ; Cerebrovascular Disorders ; blood ; epidemiology ; pathology ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Sex Distribution ; Uric Acid ; blood ; Young Adult
7.Effect of Xiaopi prescription (消痞方) on mRNA expression of c-kit in the rat with diabetic gastroparesis
Biao MU ; Zhu-Qiu QU ; Zhi-Wu LIU ; Hai-Mo CUI ; Yi-Nan QIN ; Xiang-Ming XIONG ; Ze-Bin YANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2006;0(04):-
Objective To investigate the possible role of interstitial cell of Cajal (ICE)in the pathogenesis of diabetic gastroparesis and the protective effect of"Xiaopi prescription (消痞方)".Methods Fifty healthy male Spregue-Dawley (SD) rats were randomly divided into 5 groups (each n=10):normal, model,and 3 Xiaopi prescription groups:low,middle and high dosages.Diabetes was induced by intravenous injection of alloxan,and equal amount of normal saline was intravenously injected in the normal group.Gastric lavage method was used to administer the traditional Chinese medicine decoction of Xiaopi prescription in corresponding amount (5,10 and 20 g?kg~(-1)?d~(-1)) in respective low,middle and high dosage groups.In the normal control group and diabetic model group,only equal amount of normal saline was administered into the stomach.Gastric emptying rate was measured by method of nutritious semisolid paste;c-kit positive cells of ICC were quantitatively measured with immunohistochemistry assay and computer image analysis system;c-kit mRNA positive cells were quantitatively measured with in situ hybridization and computer image analysis system.Results①Gastric emptying rate:The rate was significantly lower in the model group than that in the normal control group (P0.05),but higher than those in the model group and the low dosage group (all P0.05).②c-kit immmunohistochemistry:c-kit positive cell presented yellow in color,and its membrane was stained yellow,this kind of cells primarily were distributed around the neural plexus in the inter-space between the circular and longitudinal muscular fibers, and around the ganglionic cells forming"sheath-like"structure.The results of numbers of c-kit positive cells in the various groups:the number of the cells in the model group was significantly lower than that in the normal group (P0.05),but the numbers in the former two dosage groups were obviously higher than those in the model group (all P0.05),being significantly lower than that in the normal group,middle and high dosage groups (all P0.05),but obviously higher than those in the model group (all P0.05),being significantly lower than that in the normal,middle and high dosage groups (all P
8.Pulmonary perfusion imaging combined with Wells score and D-dimer assay for the diagnosis of acute pulmonary embolism
Chen, LIU ; Wei, FANG ; Feng, WANG ; Chang-ming, XIONG ; Zhi-hui, ZHAO ; Qing, GU ; Zuo-xiang, HE
Chinese Journal of Nuclear Medicine 2011;31(1):42-45
Objective To investigate the value of the Wells score and D-dimer assay in assisting pulmonary perfusion imaging (PPI) for the diagnosis of acute pulmonary embolism (APE). Methods One hundred twenty-one patients with suspected APE were studied from January, 2006 to December, 2008. All patients underwent the Wells score, the quantitative D-dimer assay, chest X-ray photography, and PPI. The diagnostic sensitivity, specificity, positive predictive value and negative predictive value of PPI with the assistance of Wells score and D-dimer assay were calculated. Results Fifty (41.3%) patients were diagnosed with APE. PPI combined with chest X-ray photography (Q/X scan) showed positive results in 49 patients. The sensitivity, specificity, positive predictive value and negative predictive value of the Q/X scan were 86.0% (43/50), 91.5% (65/71), 87.8% (43/49) and 90.3% (65/72), respectively. With assistance of Wells score >4 and D-dimer≥0. 5 mg/L, Q/X scan had a positive predictive value of 100.0% (29/29), for patients with Wells score ≤4 and D-dimer<0.5 mg/L, the negative predictive value for Q/X scan was 100.0% (41/41). Conclusion Combined with Wells score and D-dimer assay, PPI can make accurate diagnosis of APE.
9. Application of ICG fluorescence imaging in surgical treatment of recurrent hepatocellular carcinoma
Xiong-feng ZOU ; Zhi-xiang JIAN
Chinese Journal of Practical Surgery 2019;39(10):1040-1041
The recurrence rate of primary hepatocellularcarcinoma is very high,and repetitive resection is still the besttreatment for operable recurrent hepatocellular carcinoma.Indocyanine green(ICG) has been widely used andpopularized in resection of hepatocellular carcinoma,providesmore possibilities and diversity for the surgical treatment ofrecurrent hepatocellular carcinoma. Combined withpreoperative imaging,three-dimensional imaging technologyand intraoperative ultrasound,ICG can effectively improve theradical and accuracy of the operation for recurrent hepatocellular carcinoma. ICG-guided hepatectomy isappropriate for the new concept and technical system of liversurgery advocated by modern medical model, and realizesfunctional and anatomical hepatectomy for recurrenthepatocellular carcinoma.
10.Effects of celecoxib combined with radiotherapy on apoptosis of CNE-2Z cell lines.
Yi-zhou XIANG ; Lian-zhi WEI ; Lin YU ; Ping GUI ; Ping PENG ; Zhi-jie XIONG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2011;46(7):572-575
OBJECTIVETo investigate the effects of celecoxib combined with radiotherapy on apoptosis of CNE-2Z cell lines and the potential mechanisms.
METHODSFour groups were used, a control, celecoxib (25 micromol/L celecoxib), irradiation (8 Gy X ray) and celecoxib plus irradiation. The radiosensitising effect was detected by clone formation experiment. Flow cytometry was used to detect the apoptosis rate of cells. The expressions of Bcl-2 and Bax were assessed by immunocytochemistry. Western blot was used to examine the expression of Caspase-3.
RESULTSCelecoxib enhanced the radiosensitivity of CNE-2Z cells. In experimental group, the mean surviving fraction and the mean lethal dose of CNE-2Z cells were 0.50 and 2.36 respectively. Compared with the irradiated group, there was significant differences between the two groups (P < 0.01). Celecoxib combined with radiotherapy up-regulation the expression of Bax. The score of the expression of Bax in the control group and the experimental group were 1.221 +/- 0.116 and 2.758 +/- 0.256 respectively. Celecoxib combined with radiotherapy could inhibit the expression of the protein of Bcl-2. The score of the expression of Bcl-2 in the control group and the experimental group were 2.559 +/- 0.144 and 1.253 +/- 0.114 respectively, with significant differences (P < 0.01). Celecoxib combined with radiotherapy could increase the apoptosis rate of tumor cells with significant differences (F = 7.63, P < 0.01). Western blot showed that the expression of Caspase-3 was strengthened.
CONCLUSIONCelecoxib combined with radiotherapy could induce apoptosis and enhance the radiosensitivity of human nasopharyngeal carcinoma CNE-2Z cell lines.
Apoptosis ; drug effects ; radiation effects ; Carcinoma ; Caspase 3 ; metabolism ; Celecoxib ; Cell Line, Tumor ; drug effects ; radiation effects ; Humans ; Nasopharyngeal Neoplasms ; pathology ; therapy ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Pyrazoles ; pharmacology ; Radiotherapy ; Sulfonamides ; pharmacology ; bcl-2-Associated X Protein ; metabolism