1.Cloning Chitinase Gene of the Entomopathogene Fungus Metarhizium anisopliae HN1 and High-level Expression in Escherichia coli
Wen-Bin REN ; Shi-Qing ZHANG ; Jun-Sheng HUANG ;
China Biotechnology 2006;0(07):-
Chitinases genes from Metarhizium anisopliae which is an important entomopathogenic fungus were considered one of the key factors to invade their hosts. One Metarhizium anisopliae HN1 strain was isolated and screened. A chitinase gene was amplified by RT-PCR from Metarhizium anisopliae HN1, The whole length of this gene was 1275bp,and the nucleotide sequence of the gene was 96% similarity to that of the M. anisopliae E6 accessed in GenBank ( AF02749). The gene has been registered in GenBank and its accession number is DQ011865. The gene was subcloned into prokaryon expression vector pET-22b( + ), transformed this recombinant expression plasmid into E. coli strain BL 21 and effective expressed. The SDS-PAGE analysis indicated that the recombinant protein was 42kDa which is same to the reported article. The expression level of recombinant protein was about 63. 3% of whole expressed proteins , And when recombinant E. coli were crushed by freeze and supersonic wave , the activity assay indicates that the chitinase expressed in bacteria possesses biological activity.
4.Effects of xinshuai mistura on plasma angiotensin II, atrial natriuretic peptide and N terminal pro-brain natriuretic peptide in patients with heart failure.
Mei DAI ; Qing-xiang WEN ; Jun-ren HE
Chinese Journal of Integrated Traditional and Western Medicine 2006;26(10):888-891
OBJECTIVETo observe the effects of Xinshuai Mistura (XM) on plasma angiotensin II (Ang II), atrial natriuretic peptide (ANP) and N terminal pro- brain natriuretic peptide (Nt-proBNP) in patients with heart failure (HF).
METHODSFifty-nine patients with HF (NYHA class II-IV) were randomly divided into two groups, the treated group treated with XM and the control group with Wuling Pulvis (WP), both on the base of captopril treatment. The treatment course was two weeks. Plasma levels of Ang II , ANP and Nt-proBNP were observed.
RESULTSThe plasma levels of Ang I, ANP and Nt-proBNP showed an ascending tendency along with the increasing in severity of HF(P < 0.01), and significantly decreased after treatment in the treated group (P < 0.01), but with no remarkable difference as compared with those in the control group (P > 0.05).
CONCLUSIONXM had definite therapeutic effects on excessive neuroendocrine activation in HF patients.
Adult ; Aged ; Angiotensin II ; blood ; Angiotensin-Converting Enzyme Inhibitors ; therapeutic use ; Atrial Natriuretic Factor ; blood ; Captopril ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Heart Failure ; blood ; drug therapy ; Humans ; Male ; Middle Aged ; Natriuretic Peptide, Brain ; blood ; Peptide Fragments ; blood ; Phytotherapy ; Single-Blind Method ; Treatment Outcome
6.Effect of 3 kinds of Banxiaxiexin Decoction and its dismantlements on the amount of gastric mucus in rats
Yan-Qing GAO ; Yin-Chu SI ; Jing-Sheng HANG ; Ren-Du WEN ; Niuxin ;
Chinese Traditional Patent Medicine 1992;0(01):-
AIM:To investigate the influence of pinellia heart-draining Decoction, Licorice heart-draining Decoction, Fresh ginger heart-draining Decoction and its dismantlements on the amount of gastric mucus in rats. METHODS:20 different combination of Banxiaxiexin Decoction and its analogus according to the unform design layout, the amount of gastric mucus in rats was determinated. Stepwise regression analysis was adopted to estimate the relationship between the pharmacological data and compatibility. RESULTS:Rhizoma Coptidis could reduce the amount of gastric mucus remarkbably, the interaction of Rhizoma Pinelliae、Fructus Ziziphi Jujubae and Radix Glycyrrhizae could increase the amount of gastric mucus, and interaction was confirmed among different medicinal ingredients in prescription. CONCLUSION:Uniform design provides a referable exeperimental design for studying analogus decoction compatibility law.
7.The correlation of CT perfusion imaging with microvessel density and vascular endothelial growth factor in hepatic alveolar echinococcosis
Jing WANG ; Bo REN ; Wenya LIU ; Hao WEN ; Song QING ; Weidong XIE ; Yajing SUN ; Haitao WANG
Chinese Journal of Radiology 2011;45(11):1036-1039
Objective To explore the correlation of CT perfusion imaging with microvessel density (MVD) and vascular endothelial growth factor (VEGF) in hepatic alveolar echinococcosis (HAE).Methods Multi-slice spiral CT perfusion imaging was performed in 27 patients with HAE.Time-density curves(TDC) of the HAE peripheral area was drawn from the region of interest (ROI) with perfusion functional software.CT perfusion parameters including blood flow ( BF ),blood volume ( BV ),mean transit time(MTT) and permeability surface area product (PS) were calculated.MVD and VEGF expression of pathological specimens were examined by immunohistochemical staining with anti-CD34,anti-VEGF monoclonal antibody.The CT perfusion parameters,MVD and VEGF were compared in different types of TDC with t test.The correlation of CT perfusion parameters with MVD and VEGF were analyzed with Spearman test.Results In this group,21 cases which TDC lower than that of the liver were classified as type Ⅰ,the others 6 cases TDC higher than the liver were of type Ⅱ TDC.TDC perfusion parameters of the two types were as follows,BF:( 111.7 + 27.6),( 158.9 + 39.5 ) ml · 100 g- 1 · min - 1,BV:( 15.1 + 6.2),(26.8+8.4) ml/100 g,MTT:(7.0+4.4),(7.7+3.1) s,PS:(51.7 +17.3),(51.0+20.5) ml·100 g-1 · min-1.The significant differences of BF,BV and MVD[ (20.5 +5.4)/HP,(37.2 ±7.5)/HP,respectively ] were found between two types ( t =- 7.897,- 18.783,- 5.223,P < 0.05,respectively).There were no significant differences in MTT,PS and VEGF expression(2.1 ± 1.0,3.2 ± 1.0,respectively)between two types of TDC(P >0.05).The correlation was found between the MVD and BF and BV in the type Ⅱ TDC group( r =0.789 and 0.878,respectively) and no correlation was found between MVD and each CT perfusion parameters in the type Ⅰ TDC group ( P > 0.05 ).There was no correlation between the VEGF expression and CT perfusion parameters in two types of TDC ( P > 0.05 ).Conclusion CT perfusion imaging with different type of TDC reflected different situation of angiogenesis in HAE peripheral area,which could be a potential technique to illustrate the microcirculation of this disease.
8.Cytochrome C oxidase subunit 1 mutation result in mitochondrial encephaiomyopathy with lactic acidosis and stroke-like episodes
Jin-Song JIAO ; Yong-Qing ZHANG ; Hao-Ping DU ; Kang WANG ; Ren-Bin WANG ; Guo-Xiang WANG ; Wen HONG
Chinese Journal of Neurology 2000;0(04):-
Objective To report a 33-year-old man with post-headache stroke-like episodes,with whom ischemic changes were found in basal ganglia and occipital-temporal lobe and muscle biopsy revealed abnormal mitochondrial structure and function without regular mutations detected in mtDNA.Methods Gene chip technique was used to detect the mutation of whole sequence of mtDNA,and direct sequencing technique was used to confirm the mutations.Results Three mutations were found.A new mutation in the mitochondrial cytochrome C oxidase subunit 1 (MTCO1),a T→C transition at nucleotide position 6253 resulted in conservative methionine transferring to threonine.His mother also held the mutation,which was not found in 98 control samples.So T6253C was considered the nosogenetic mutation.Conclusion This is the first time to report a mutation in MTCO1 responsible of MELAS.
9.Application of a simple random sampling method on surveys at the community level
Jun LV ; Qing-Min LIU ; Yan-Jun REN ; Wen-Feng WANG ; Ting GONG ; Li-Ming LI
Chinese Journal of Epidemiology 2010;31(4):421-423
Objective To study the feasibility of a simple random sampling on surveys at the community level and to evaluate the quality of samples under survey.Methods A simple random sample of households was taken,based on the electronic listings of community households from Gongshu and Xiacheng districts of Hangzhou city.One of the adults aged 18 to 64 years in the sampled households was identified with KISH method to finish a questionnaire survey.More than 500 people from the sample size was required in each district.Results Of 950 sampled households in Xiacheng district,511 (53.8%) finished the survey while 506 (36.7%) out of the 1380 sampled households in Gongshu district did.The proportions of non-response due to the following reasons as:none with eligible age in the households,relocation of the original household,mass relocation of the community,and errors in the household listings etc.were 38.3% and 43.5% respectively,in the two districts.Proportions attributed to non-response and refusal to response of sampled household or individual were 8.0% and 19.9% respectively.No statistical significant differences in age and gender were found between the surveyed samples and the population in the sampled households,or in gender between the populations in the sampled households and in Hangzhou city.However,the population in the sampled households showed a more aging population structure than the population in Hangzhou city.Conclusion In a geographically limited area,using a simple random sampling method to do the survey is feasible,based on the electronic listings of household.Enough time spent during the household visit guarantees the interviewers to get a representative sample of the sampling frame.There is an urgent need for the timeliness,completeness and accuracy of electronic household listings to be improved.
10.Mechanism of bradykinin on inflammations of retinal pigment epithelium cells
Wen-Ting, CAI ; Cheng-Da, REN ; Qing-Yu, LIU ; Qing-Quan, WEI ; Ya-Ru, DU ; Qian-Yi, WANG ; Jun-Ling, LIU ; Meng-Mei, HE ; Jing, YU
International Eye Science 2016;16(8):1430-1434
Abstract?AIM: To investigate mechanism of bradykinin ( BK) on inflammations of retinal pigment epithelium ( RPE) cells.?METHODS: ARPE -19 cells were cultured in vitro, stimulated by 100nM BK for 24h. Cell morphology changes were observed by microscope, and BK receptor localization was detected through cell immunofluorescence. Changes of Ca2+in BK and BR antagonist stimuli were detected by laser scanning confocal microscopy.The expressions of COX-1, COX-2, eNOS and iNOS protein in control group and BK group were detected by Western Blot.?RESULTS: After the stimulation of BK, there was no significant changes of ARPE-19 cells in morphology.Kinin B1 receptors ( B1R ) and B2 receptors ( B2R ) could be detected in ARPE-19 cells.Compared with control group, Ca2+concentrations significantly increased in BK group; in B1R antagonist group and B2R antagonist group Ca2+concentrations increased less than BK group; B1R and B2R antagonist group showed no obvious changes in Ca2+concentrations.Compared with control group, COX-2 and iNOS protein concentrations were significantly increased in BK group (P<0.001).?CONCLUSION:BK induces the increasing expression of COX-2 and iNOS in the cultured ARPE cells through binding with either B1R or B2R.