7.A study on antiviral activity of defensins from neutrophil of human and rabbit against SARS coronavirus in vitro
Yongjie WANG ; Xiaoyu LI ; Ming QI
Medical Journal of Chinese People's Liberation Army 2001;0(12):-
Objective To determine the anti-SARS coronavirus (SARS-CoV) activity of defensins from human and rabbit, and explore the effects of defensin during the onset and progression of SARS. Methods Defensins were purified from neutrophil of human and rabbit respectively, namely HNP (human neutrophil protein) 1-3 and RNP (rabbit neutrophil protein) 1-5, by acid-urea-polyacrylamide gel electrophoresis (CAU-PAGE). The effect of defensin on Vero-E6 cell infected by SARS CoV BJ01 strain as well as the replication of virus were observed by micro-culture technique. Rusults The HNP 1-3 and RNP 1-5 separated by CAU-PAGE presented a single band with molecular weight of approximately 4000 dalton on 15% SDS-PAGE. HNP 1-3 was identified and validated by quantitative ELISA. The activity of anti-escherichia coli (ATCC25922 strain) of HNP 1-3 and RNP 1-5 was tested by MIC determination method, and the IC50 values obtained were 25~50?g/ml and 5~10?g/ml, respectively. In the antiviral experiment, HNP 1-3 and RNP 1-5 in the concentration from 25 to 100?g/ml were not able to prohibit cytopathic effect (CPE) of SARS-CoV and the replication of virus in Vero-E6. Conclution The natural defensins (HNP 1-3 and RNP 1-5) purified from neutrophils do not play any prohibitory or eliminative effects on SARS-CoV, indicating that that human and rabbit do not have natural immunity against SARS-CoV.
8.An ELISA for detection of anti-dsDNA by using plasmid DNA as antigen
Hua XIONG ; Xiaojun LI ; Ming QI
Chinese Journal of Laboratory Medicine 2000;0(06):-
Objective To develop an ELISA for detection of anti-dsDNA antibody by using plasmid DNA as antigen.Methods DNA in plasmid pBV220 for prokaryotic expression vector was purified by base-cleavage. The microtiterplates were pretreated by poly-L-lysine and coated by the plasmid DNA in a dilution of 1∶50 as antigen. An ELISA method for detection of serum anti-dsDNA antibodies was developed with HRP-SPA as enzyme-labeled marker.The IIF using crithidia lucilia as substrate was performed simultaneously for comparison. The serum samples from 64 patients with SLE, 8 with MCTD and 17 with RA were detected. Results The concentration of DNA was 1 54 g/L by UV spectrophotometer at wavelength of 260 nm. The positive percentage of ELISA for anti-dsDNA was higher than that of IIF. By comparison with IIF the positive percentages in SLE, MCTD and RA groups were 23 4% vs 17 2%, 12 5% vs 12 5% and 11 8% vs 5 9%, respectively, and the coincident rates between the 2 methods were 93 8%, 100% and 94 1% respectively. The sensitivity and specificity of the developed ELISA for detection of anti-dsDNA were 100% and 93 4% when IIF was as gold standard.Conclusion The ELISA by using plasmid DNA as antigen to detect anti-dsDNA has fine precision, sensitivity and specificity. Its positive rate is higher than that of IIF thus it will contribute to monitor the activities for SLE patients′ condition.
9.Association study on classification,injury score, kyphosis deformity and nerve function injury of thoracolumbar fracture
Ming CHENG ; Yue WANG ; Qi LI
Chongqing Medicine 2017;46(16):2198-2199,2203
Objective To investigate the association between the lumbar vertebral fracture damage degree with the fracture classification,injury score,kyphosis deformity and nerve function injury.Methods According to the damage degree of posterior lig ament complex(PLC),the patients were divided into the PLC intact group and PLC injury group.Its relationship with PLC injury was researched by evaluating the fracture classification,injury score and nerve function injury situation in the two groups.Results The LCS score and TLICS score in the PLC injury group were (7.1±0.8) points and (8.2±0.6) points,which were higher than (5.7±0.5) points and (4.6±0.7) points in the PLC intact group.The Denis score in the PLC injury group was more serious.The Cobb angle in the PLC injury group was 29°,and which in the PLC intact group was 19°.The proportion of nerve function insufficiency in the PLC injury group was 89%,while which in the PLC intact group was only 60 %.Conclusion The thoracolumbar vertebral fracture is closely related with PLC.PLC damage degree is positively correlated with the fracture classification,injury score,kyphosis deformity and nerve function injury degree.
10.Determination of Oleic Acid and Linoleic Acid in Brucea javanica Oil by RP-HPLC
China Pharmacy 2007;0(36):-
OBJECTIVE:To establish an RP-HPLC method for determination of the contents of oleinic acid and linolic acid in Brucea javanica oil. METHODS:The derivative fatty acid was obtained from extracting and separating process using ?-bromo-acetophenone as derivative reagent,K2CO3 as base and N,N-dimethylformamide (DMF) as solvent. The sample was separated on Shim-pack VP-ODS column(150 mm?4.6 mm,5 ?m); the detection wavelength was set at 242 nm. The mobile phase consisted of methanol-acetonitrile-water(30∶20∶50). RESULTS:The linear ranges of oleinic acid and linolic acid were 0.025~1.0 mg?mL-1(r=0.999 6) and 0.1 ~ 0.8 mg?mL-1(r=0.999 0),respectively. The average recoveries were 98.54% and 98.50%,respectively,and their RSD were 1.81% and 1.90%,respectively (n=6 each). CONCLUSION:The method is accurate and specific,and it is applicable for the quality control of B.javanica oil.