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Objective:To explore the distribution characteristics of respiratory pathogens in patients with community-acquired pneumonia in Lianyungang.Methods:A total of 612 patients admitted to the second people′s Hospital of Lianyungang City because of community-acquired pneumonia (CAP) in 2019 were selected as subjects. Sputum or pharyngeal swabs were collected to extract nucleic acids, and 13-fold nucleic acids of respiratory pathogens were detected by PCR capillary electrophoresis fragment analysis. SPSS statistical software and GraphPad5.0 statistical mapping software were used for statistical analysis.Results:The physical examination rate of respiratory pathogens in the adult group was 82.0% in winter, 48.4% in spring, 28.0% in autumn, 20.0% in summer, χ 2=38.473, P=0.000. The positive rate of nucleic acid detection was significantly different in different seasons, among which the physical examination rate of respiratory pathogens in winter was the highest. The physical examination rate of respiratory pathogens in the juvenile group was 86.0% in spring, 76.2% in winter, 71.3% in summer and 66.7% in autumn, χ 2=7.946, P=0.047 . The positive rate of nucleic acid detection was calculated according to gender grouping. The comparison of nucleic acid positive rate between adult group and juvenile group in different seasons: 86.0% vs 48.4% in spring, χ 2=19.436, P=0.000; 71.3% vs 20.0% in summer, χ 2=22.180, P=0.000; 66.7% vs 28.0% in autumn, χ 2=13.485, P=0.000; 76.2% vs 82.0% in winter, χ 2=0.758, P=0.384. Except in winter, the detection rate of nucleic acid of pathogens in the juvenile group was significantly higher than that in the adult group. Conclusions:The nucleic acid detection rate and etiological distribution characteristics of respiratory pathogens are different in patients with community-acquired pneumonia in different seasons and different age groups. 13 kinds of multiple detection methods of respiratory pathogens can provide favorable laboratory data support for the diagnosis and treatment of clinical CAP patients.
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The blood samples of 294 Chinese Han patients with type 1 diabetes mellitus(T1DM) and 199 controls were collected and their genomic DNAs were extracted. The single nucleotide polymorphisms rs17534481, rs12095738, rs2988276, and rs6668182 of CD247 gene were detected. The results showed that rs17534481 polymorphism of CD247 gene was associated with T1DM in Chinese Han population, and T allele was a protective factor for type 1 diabetes mellitus( OR=0.667, P=0.037). There was no significant difference in genotype frequency and allele frequency of rs12095738, rs2988276, and rs6668182 between type 1 diabetes group and control group( P>0.05).
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Objective The primary culture of synovial fibroblasts is a convenient tool to study the pathology and physiology of synovial tissues .An improved method was constructed in this study by C57BL /6 mice to study the mechanism of rheumatoid ar-thritis(RA) .Methods The synovium around the hip joints were collected .Attention should be paid to eliminate the egg-yolk like yellow oval substance in the middle of the synovium .The synovium was transferred into a 1 .5 mL Eppendorf tube containing 0 .5%type Ⅳ collagenase and cut into 1 mm3 blocks or so .The Eppendorf tube was placed in 37 ℃ Constant temperature orbital shaker incubator for 60 min .After digestion ,the tube was placed on the Vortex for a high-speed oscillation for 1 .5 minutes to guarantee the separation of cells .Results Within about 1 week ,the first passage was performed by the trypsin digestion method .On day 10 , the number of synovial macrophages reached the maximum and then decreased gradually .After the third generation (day 15 to 20) , the synovial macrophages generally disappeared .Vimentin was suitable for the immunofluorescence cytochemical staining for the synovial fibroblasts .The cell purity was indicated as > 95% .The cytometric analysis indicated that purity of Vimentin and CD90 .2-labelled cells was over 95% ;the purity of CD54-labelled cells was 80% approximately .Conclusion It is a simple and effective method for primary culture of synovial fibroblasts in mice .
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<p><b>OBJECTIVE</b>To investigate the effect of tumor necrosis factor-α (TNF-α) on the release of matrix metalloproteinase-3 (MMP-3), MMP-9, and interleukin-17 (IL-17) in cultured mouse bone marrow-derived mast cells (BMMCs) in vitro.</p><p><b>METHODS</b>Primarily cultured mouse BMMCs at 8 weeks were exposed PBS (control) or TNF-α at the concentrations of 2, 10, or 50 ng/mL for 12 or 24 h. Real-time PCR was performed to detect the mRNA expressions of MMP-3, MMP-9, and IL-17 in the exposed cells.</p><p><b>RESULTS</b>A 12-hour exposure of the BMMCs to TNF-α caused significantly increased expressions of MMP-3, MMP-9, and IL-17 in a concentration-dependent manner (P<0.05). Prolonged exposures of the cells to 2 and 10 TNF-α for 24 h further increased MMP-3, MMP-9, and IL-17 mRNA expressions, but exposure to 50 ng/mL TNF-α for 24 h increased only MMP-3 and MMP-9 expressions but not IL-17 mRNA expression.</p><p><b>CONCLUSIONS</b>TNF-α treatment of primarily cultured BMMCs can significantly increase the cellular expressions of MMP-3, MMP-9, and IL-17 mRNA in a time- and dose-dependent manner.</p>