1.Effects of methylprednisolone on expression of MMP-9 and airway inflammation in murine models of asthma
jian, ZHANG ; jian-ming, ZHU ; jian-wen, BAI ; min-jia, LIN ; shu-min, XU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(05):-
Objective To investigate the effects of methylprednisolone(MP) on the expression of matrix metalloproteinase(MMP)-9 and airway inflammation in murine models of asthma. Methods Thirty female BALB/c mice were randomly divided into asthma group,MP group and control group(n=10).Murine models of acute asthma were established by ovalbumin(OVA) via peritoneal injection and intranasal instillation.The pathological changes of lung tissues were observed with HE staining,and cell quantitation was conducted in bronchalveolar lavage fluid(BALF).The expression of MMP-9 protein was determined by immunohistochemistry and gelatin zymogram,and the expression of MMP-9 mRNA was detected by RT-PCR. Results Compared with control group,there were more significant airway spasm and more infiltration of inflammatory cells in histologic examination,and there was higher eosinophil cell quantitation in BALF in asthma group(P
2.Expression of Lipase Genes in Bacillus subtilis and Characterization of the Recombinant Lipase
Jian-Bo JIA ; Xiang-Qian LI ; Min HU ;
China Biotechnology 2006;0(01):-
A Staphylococcus aureus JH strain which can produce lipase was obtained from environment.According to polysequence comparision of prokaryote’s lipase gene published on NCBI,its sequence is very conservative. Lipase gene was obtained by PCR from genome DNA of Staphylococcus aureus JH,and then it was incorporated into plasmid pC194 and transformed into B.subtilisH11 .The recombinant lipase precipitated by(NH4)2SO4, purified by ion exchange chromatography and was identified by SDS-PAGE.It was revealed that the molecular mass of the recombinant lipase is 32kDa;the recombinant lipase show maximum activity at 41℃,pH8.0;the values of Km and Vm were found to be 0.34mmol/L and 308?mol/mg.min;The lipase can be activated by the metal ions Ca2+,K+ and Mg2+ and be inhibited by Fe2+,Cu2+,and Co2+.
6.Expression of P-MLCK in human pulmonary arterial endothelial cell induced by lipopolysaccharide
min-jia, LIN ; jian-wen, BAI ; jin-shi, LI ; shu-min, XU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(04):-
Objective To study the expression of phosphorylated myosin light chain kinase(P-MLCK) in human pulmonary arterial endothelial cell(HPAEC) induced by lipopolysaccharide(LPS). Methods HPAECs were cultured in vitro and treated with LPS(2 ?g/mL) and normal saline for 1 h,respectively.Immunofluorescence method and western blotting were used to detect P-MLCK. Results Compared with normal saline group,the number of HPAECs decreased,but the morphology of cells did not change.After treatment of LPS for 30 and 60 min,the expression of P-MLCK in HPAEC increased from 0.41?0.05 to 0.82?0.43 and 1.56?0.07,respectively(P
8.Antimicrobial resistance features and molecular typing of clinically isola-ted m ethicillin-resistant Staphylococcus aureus
Min JIA ; Yuan-Shan JIANG ; Jian-Hua ZHU ; Jia-Jia GAO ; Yong-Tao WANG ; Zhi-Min HU ; Zhi-Gang LIU
Chinese Journal of Infection Control 2018;17(4):289-293
Objective To study antimicrobial resistance and genotyping of methicillin-resistant Staphylococcus au-reus(MRSA). Methods A total of 967 no-repetitive strains of Staphylococcus aureus(S.aureus)isolated from a hospital between January 2014 and November 2015 were collected,antimicrobial susceptibility testing,mecA gene,and Panton-Valentine leukocidin gene(PVL gene)were detected;staphylococcal cassette chromosome mec(SCCmec)typing,multilocus sequence typing(MLST),S.aureus protein A(spa)gene typing,and S.aureus ac-cessory gene regulator(agr)typing were performed with multiplex polymerase chain reaction. Results Of 967 strains of S.aureus,210(21.72%)were MRSA;detection rate of MRSA from sputum specimen was higher than that of skin and soft tissue specimen(68.09% vs 1 1.83% ,P<0.05);vancomycin- and linezolid-resistant S.aureus strains were not found,susceptibility rates of MRSA to gentamicin,tetracycline,erythromycin,clindamycin,levo-floxacin,ciprofloxacin,moxifloxacin,nitrofurantoin,and rifampicin were all lower than those of methicillin-sensi-tive Staphylococcus aureus(MSSA),differences were all statistically significant(all P<0.05);antimicrobial sus-ceptibility rate of MRSA to compound sulfamethoxazole was higher than MSSA,difference was significant(P<0.05). Susceptibility rates of MRSA isolated from skin and soft tissue to gentamicin,levofloxacin,ciprofloxacin,moxifloxacin,and rifampicin were 86.90% -95.24%,while MRSA isolated from sputum were only 1.56% -15.63%.Of 967 strains of S.aureus,210 harbored mecA gene,10 harbored PVL gene,8(3.81%)of 210 MRSA strains weren't typed. The main types of MLST,SCCmec,spa,and agr were ST 239(n= 177 strains),type Ⅲ(n= 177 strains),t 030(n= 177 strains),and typeⅠ(n= 196 strains)respectively.Conclusion The main epidemic clone of MRSA strain in this hospital is ST239-MRSA-SCCmec III-t030,antimicrobial resistance is serious,monitoring on drug-resistant strains in hospital should be strengthened.
9.Expression of myosin light chain kinase in acute lung injury
jin-shi, LI ; jian-wen, BAI ; min-jia, LIN ; dou-xia, ZHANG
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(12):-
Objective To study the inflammation and expression of myosin light chain kinase(MLCK) through establishing acute lung injury animal model of mice induced by lipopolysacchride(LPS), and approach the role of MLCK in the mechanism of acute lung injury.Methods Twenty female BALB/c mice were randomly divided into LPS group(n=10) and control group(n=10).The BALB/c mice of LPS and control groups were induced by 30 ?L 0.9% NaCl via intranasal instillation,while only LPS group was treated with LPS(20 ?g/each mice).The pathology,wet/dry lung weight ratio and the total cell quantitation in bronchoalveolar lavage fluid(BALF)were compared between these two groups.Furthermore,immunohistochemistry assays were used to determine the status of MLCK expression in the lung.And RT-PCR was adopted to determine the status of MLCKmRNA in the lung. Results Compared with the control group,the LPS group showed more serious pulmonary hemorrhage,edema and infiltration of neutrophils, significantly increased water content in the lungs and total cell quantitation in BALF(P
10.Effect of montelukast and dexamethasone on inflammation of asthma
min-jia, LIN ; jian-wen, BAI ; dou-xia, ZHANG ; jin-shi, LI
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(01):-
Objective To study the effect of montelukast(MK)and dexamethasone(Dex)on inflammation of asthma.Methods Asthma model was established and treated with MK or Dex.Bronchoalveolar lavage fluid(BALF) and histopathologic change were observed,IL-5mRNA of lung and bone marrow cells were detected by in situ hybridization,IL-5 immunoreactive cells by immunohistochemistry,and CD34+ and CD3+ of bone marrow cells by flow cytometry. ResultsCompared with asthma group,the number of total cells and eosinophils in BALF of MK group and Dex group were significantly decreased(P0.05). Conclusion MK and Dex can well inhibit airway inflammation and expression of IL-5mRNA in lung and bone marrow cells,though MK may be inferior to Dex in some aspects.The combined treatment of leukotriene receptor antagonist and glucocorticosteroid may be a new direction for asthma.