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1.
Acta Academiae Medicinae Sinicae ; (6): 203-207, 2019.
Article in Chinese | WPRIM | ID: wpr-776049

ABSTRACT

Objective To explore the clinical value of one-step visualization loop-mediated isothermal amplification(LAMP)in the detection of Mycoplasma pneumoniae(Mp). Methods One-step visualized LAMP,polymerase chain reaction(PCR),and enzyme-linked immunosorbent assay(ELISA)were used to simultaneously detect 108 clinical Mp specimens in children,which included 73 cases of Mp infection diagnosed by PCR and 35 cases of other chronic/acute respiratory tract infections.On the first day of admission,one-step visualization LAMP,PCR(fluorimetric method),and ELISA were used to test the throat swab and serum sample obtained from the same patient,and the Kappa value was calculated.The consistence between LAMP and PCR and that between LAMP and ELISA were compared.On the fifth day of admission,40 patients were resampled and the findings of these three tests on the first day and on the fifth day were compared. Results One-step visualization LAMP had a sensitivity of 100% and a specificity of 94.3%,whereas ELISA had a sensitivity of 65.8% and a specificity of 82.9%.The ratio of Kappa camparing one-step visualization LAMP and PCR was 0.956 and the ratio of Kappa camparing one-step visualization LAMP and ELISA was 0.38.The number of positive specimens detected by LAMP was higher than that by ELISA on the first day. Conclusions One-step visualization LAMP has excellent sensitivity and specificity in detecting early acute Mp infection.It has high consistency with PCR and can be applied to detect Mp.


Subject(s)
Child , Humans , Enzyme-Linked Immunosorbent Assay , Mycoplasma pneumoniae , Nucleic Acid Amplification Techniques , Pneumonia, Mycoplasma , Diagnosis , Polymerase Chain Reaction , Sensitivity and Specificity
2.
Chinese Journal of Hepatology ; (12): 619-623, 2013.
Article in Chinese | WPRIM | ID: wpr-278028

ABSTRACT

<p><b>OBJECTIVE</b>To study the relationship between the patatin-like phospholipase domain-containing protein 3 (PNPLA3) gene and hereditary susceptibility to non-alcoholic fatty liver disease (NAFLD) by detecting single nucleotide polymorphisms (SNPs).</p><p><b>METHODS</b>Peripheral blood DNA from 315 patients diagnosed with NAFLD (including the spectrum of simple steatosis (SS) and non-alcoholic steatosis (NASH)) and 336 control subjects was used to determine the PNPLA3 genotype by polymerase chain reaction (PCR) and direct sequencing. The relationship of SNPs and NAFLD-related markers of liver function were assessed by correlation analysis.</p><p><b>RESULTS</b>The SNP rs738409 was identified in more of the NAFLD patients (allele variant frequencies: NAFLD, 65.40%; NASH: 71.87%; SS, 56.47%) than in the controls (33.18%). Case-control analysis revealed that carriers of the 148GG genotype were at 3.81-fold (95% CI: 3.03 ~ 4.79) higher risk of developing NAFLD and at 1.97-fold (95% CI: 1.41 ~ 2.75) higher risk of progressing from SS to NASH, compared with non-carriers. rs738409 was also found to be associated with serum levels of alanine aminotransferase (ALT) and y-glutamyltransferase (y-GT) (both P less than 0.05). Carriers of the 148GG genotype had significantly higher body mass index, ALT, and fasting insulin than carriers of the 148CC genotype (all P less than 0.05), and significantly higher level of serum HDL than carriers of either the 148CC genotype or the 148GC genotype (both P less than 0.05).</p><p><b>CONCLUSION</b>Polymorphisms in the PNPLA3 gene may play an important role in mediating susceptibility to developing NAFLD in the Chinese population. The rs738409 polymorphism, in particular, is related to development and progression of NAFLD and may play a role in the contribution of PNPLA3 to NAFLD pathogenesis.</p>


Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Alleles , Case-Control Studies , Fatty Liver , Genetics , Gene Frequency , Genetic Predisposition to Disease , Genotype , Lipase , Genetics , Membrane Proteins , Genetics , Non-alcoholic Fatty Liver Disease , Polymorphism, Single Nucleotide
3.
Acta Pharmaceutica Sinica ; (12): 737-740, 2009.
Article in Chinese | WPRIM | ID: wpr-278190

ABSTRACT

This study is to investigate the effect of ibudilast on apoptosis of airway eosinophil in asthmatic guinea pigs and its mechanism. Experimental asthma model of guinea pigs was induced with ovalbumin (OVA). Differential count in BALF was examined. The apoptosis of eosinophils (EOS) was labeled with TdT-mediated dUTP nick end labeling (TUNEL) technique. Fas mRNA expression of EOS was detected by reverse transcription-polymerase chain reaction (RT-PCR). The quantification of GM-CSF and IL-5 in BALF was conducted with ELISA. After treatment of ibudilast, the number of EOS and the quantification of GM-CSF and IL-5 decreased significantly. The number of apoptotic cells as well as Fas mRNA expression of EOS obviously increased. The results indicated that anti-asthma mechanisms of ibudilast can antagonize asthma through decreasing the number of EOS, inducing apoptosis of EOS, enhancing Fas mRNA expression of EOS and reducing the content of GM-CSF and IL-5.


Subject(s)
Animals , Female , Male , Anti-Asthmatic Agents , Pharmacology , Apoptosis , Asthma , Metabolism , Pathology , Bronchoalveolar Lavage Fluid , Eosinophils , Cell Biology , Metabolism , Granulocyte-Macrophage Colony-Stimulating Factor , Metabolism , Guinea Pigs , Interleukin-5 , Metabolism , Pyridines , Pharmacology , fas Receptor , Metabolism
4.
Genomics, Proteomics & Bioinformatics ; (4): 51-60, 2008.
Article in English | WPRIM | ID: wpr-316998

ABSTRACT

To further study the characteristics of changes on the molecular level of rice mutants induced in space environment, we analyzed proteins in leaves and seeds of four rice mutants (two high-tillering and two low-tillering) in the 8(th) and 9(th) generations after a 15-day spaceflight, and compared with their ground controls by two-dimentional polyacrylamide gel electrophoresis and reverse phase liquid chromatography (RPLC). In addition, the albumin, globulin, prolamine, glutelin, and amylose of the mutant seeds were analyzed by RPLC and ultra-violet spectrometry. The results showed that the low-abundance proteins of leaves in the peak tillering stage are more likely to be induced compared with their corresponding controls. The albumin, globulin, and prolamine of the mutant seeds revealed changes when compared with their controls, and the characteristics of changes in different mutants were stably inherited in the 8(th) and 9(th) generations, suggesting that they can be used as bio markers to identity the mutants induced by spaceflight. Moreover, two proteins (SSP9111 and SSP6302) were found to be expressed with high intensity (two-fold change) in different mutants, which were both correlated with photosystem according to mass spectrometry and database searching.


Subject(s)
Albumins , Genetics , Metabolism , Amylose , Genetics , Metabolism , Chromatography, Liquid , Electrophoresis, Gel, Two-Dimensional , Extraterrestrial Environment , Gene Expression Regulation, Plant , Genetic Variation , Genomic Instability , Globulins , Genetics , Metabolism , Mass Spectrometry , Mutation , Oryza , Genetics , Metabolism , Plant Leaves , Genetics , Plant Proteins , Genetics , Metabolism , Prolamins , Seeds , Genetics , Space Flight
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