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Objective:To analyze the clinical features of patients with COVID-19 in Chongqing Municipality.Methods:The clinical data, laboratory tests and chest imaging findings of 153 patients COVID-19 admitted in Chongqing Public Health Medical Center from January 26 to February 5, 2020 were retrospectively reviewed. According to the relevant diagnostic criteria, patients were divided into non-severe group (n=132) and severe group (n=21). The correlation between serum index changes and disease severity was analyzed.Results:The proportion of patients with underlying diabetes or chronic respiratory diseases in severe group was significantly higher than that in non-severe group ( χ2=11.04 and 6.94, P<0.05). The proportion of symptom-free patients in non-severe group was significantly higher than that in severe group ( χ2=4.09, P<0.05). The symptoms of fever, fatigue and muscle soreness in the severe group were more common than those in the non-severe group ( χ2=4.40, 14.42 and 22.67, P<0.05). Among the concomitant symptoms, the proportion of cough and shortness of breath in the severe group was higher than that in the non-severe group ( χ2=8.46 and 4.80, P<0.05). C-reactive protein and D-Dimer levels were higher in the severe group than those in the non-severe group ( Z=-4.39 and -1.96, P<0.05), and the number of CD3 + T lymphocyte cells, CD4 + T lymphocyte cells and CD8 + T lymphocyte cells in the severe group was lower than that in the non-severe group ( Z=27.25, 20.60 and 17.36, P<0.05). Compared with the non-severe group, both lungs and the right lung lower lobe were more susceptible to be involved( χ2=9.71和23.61, P<0.05). Conclusions:There are significant differences in underlying diseases, clinical symptoms, imaging manifestations and laboratory findings between severe and non-severe patients with COVID-19.
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Objective@#To analyze the clinical data of 153 patients with novel coronavirus pneumonia (COVID-19) in chongqing ,and provide reference and thinking for the diagnosis and treatment.@*Methods@#Analyze the clinical data, laboratory examination and chest imaging characteristics of 153 COVID-19 patients in Chongqing Public Health Medical Center from January 26 to February 5, 2020. According to the relevant diagnostic criteria ,patients were divided into non-severe group(n=132) and severe group(n=21),and analyze the correlation between serum index changes and disease severity.@*Results@#Combined with diabetes and chronic respiratory diseases, the severity of the disease was statistically significant (χ2=11.04和6.94, P<0.05). No symptoms were found in patients with mild illness (χ2=4.09, P<0.05) .The proportion of fever and muscle soreness in the severe group was higher than that in the non-severe group (χ2=4.40 and 22.67,P<0.05).Among the concomitant symptoms, the proportion of cough and shortness of breath in the severe group was higher than that in the non-severe group (χ2=8.46 and 4.80,P<0.05).C-reactive protein and d-dimer were higher in the severe group than in the non-severe group (t=43.44 and 37.13, P<0.05), and the number of CD3+T lymphocyte cells, CD4+T lymphocyte cells and CD8+T lymphocyte cells in the severe group was lower than that in the non-severe group (Z=27.25, 20.60 and 17.36, P<0.05).Compared with the non-severe group, both lungs and the right lung lower lobe were more susceptible to involved (χ2=6.95和20.39, P<0.05) .@*Conclusion@#Severity of COVID-19 was associated with underlying disease, symptoms, site of involvement, C-reactive protein, d-dimer, CD3+T lymphocyte count, CD4+T lymphocyte count, and CD8+T lymphocyte count.
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Objective·To investigate the bacteriostasis effect of Mycobacterium tuberculosis (MTB) polyphosphate kinase 2 (PPK2) aptamer on MTB in vitro.Methods·The bioinformatics method was used to analyze the homology of MTB PPK2 and common pathogens of respiratory tract,and the PPK2 phylogenetic tree was constructed.The binding affinity of the PPK2 aptamer to H37Rv,BCG,Mycobacterium smegmatis,Pseudomonas aeruginosa and Acinetobacter baumannii was analyzed by enzyme-linked oligonucleotide assay (ELONA).The PPK2 aptamer was incubated for 24 h in serum and its biological stability in serum was analyzed by agarose gel electrophoresis.The minimum inhibitory concentration (MIC) of the PPK2 aptamer to H37Rv was determined by micro-azure method.H37Rv was inoculated with 1 μmol/L PPK2 aptamer or random sequence on Roche culture medium for 10 d and colony growth status was observed.H37Rv was co-cultured with different concentrations of PPK2 aptamer for 10 d,absorbance at 600 nm was measured by microplate reader.The effect of PPK2 aptamer on the growth of H37Rv was observed.Results·PPK2 phylogenetic tree constructed by bioinformatics analysis showed that PPK2 protein of H37Rv was not closely related to the common pathogens of respiratory tract,and it was relatively close to Pseudomonas aeruginosa.The ELONA assay results showed that the PPK2 aptamer binded selectively to H37Rv.Agarose gel electrophoresis analysis showed PPK2 aptamer in serum was at least stable for 8 h.The MIC of the PPK2 aptamer to H37Rv was 50 nmol/L.The colony growth of Roche culture showed that PPK2 aptamer had an inhibitory effect on H37Rv growth.Growth inhibition test showed that the absorbance at 600 nm of H37Rv showed a decreasing trend with the increase of PPK2 aptamer concentration,which indicated that PPK2 aptamer had an inhibitory effect on H37Rv growth.Conclusion·PPK2 aptamer has good antibacterial activity against H37Rv in vitro.
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Objective To study pharmacodynemics and pharmacokenitics of apixaban in rats and investigate the correlation between them.Mehtods The UPLC-MS/MS method was applied to determining the plasma concentration of apixaban and draw the concentration-time curve.Meanwhile,the extension rate of prothrombin time (PT) was determined to draw the effect-time curve.Then the relationship between concentration and effect could be evaluated.Results After iv administration of apixaban (2 mg/kg) in rats,the main pharmacokinetic parameters AUC0-∞ and T1/2z were (4 016.07 ± 1 160.46) μg·h/L and (2.95 ± 1.59) h,respectively.After ig administration of apixaban (10 mg/kg),the main pharmacokinetic parameters AUC0-∞,T1/2z,Cmax,Tmax and bioavailability were (17 973.48 ± 3145.30) μg·h/L,(1.52 ± 0.36) h,(4 949.12 ± 615.38) μg/L,(1.00 ± 0.71) h and 89.5％,respectively.Apixaban (10 mg/kg) significantly increased PT and the effect lasted about 2 h.The changes of apixaban plasma concentration and PT extension rate were synchronous.Conclusion Apixaban has the characteristics of high oral bioavailability and rapid absorption.There is a significant correlation between PT extension rate and its plasma concentration after ig administration of 10 mg/kg in rats.
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Objective To establish a hanging drop 3D cell culture model of human colon cancer cell (HT29) in 48-well cell culture plate,at the same time,through the comparison of several cell viability detection methods to determine the appropriate one for this cell culture way.Methods HT29 cells of 2 375,3 164,4 218,5 625,7 500 and 10 000/well were seeded in the bottom of the 48-well culture plate to form droplets.After 2 d of inversion culture,the cell spheroids were formed and incubated in medium for another 3 d.The volume of cell spheroids were measured,and the absorbance (A) values were detected through APH assay,MTT assay,MTT assay after digestion,CCK-8 assay and CCK-8 assay after digestion.The results were compared among different methods.Results After 5 d of culture,the cell spheroids were formed perfectly at the density of 2 375-10 000/well,and the volumes were in good linear with the original cell inoculation number at the density of 2 375-7 500/well.The A values of APH assay,MTT assay after digestion and CCK-8 assay after digestion increased with the increase of cell inoculation amount;But the cell ball digestion process was complex,and the cell viability was damaged.However,the A values of MTT and CCK-8 assay increased slowly.Conclusion The method of a hanging drop 3D cell culture model in 48-well culture plate combining with APH assay to detect cell viability is economical,accurate and easy to operate.
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Objective To establish a hanging drop 3D cell culture model of human colon cancer cell (HT29) in 48-well cell culture plate,at the same time,through the comparison of several cell viability detection methods to determine the appropriate one for this cell culture way.Methods HT29 cells of 2 375,3 164,4 218,5 625,7 500 and 10 000/well were seeded in the bottom of the 48-well culture plate to form droplets.After 2 d of inversion culture,the cell spheroids were formed and incubated in medium for another 3 d.The volume of cell spheroids were measured,and the absorbance (A) values were detected through APH assay,MTT assay,MTT assay after digestion,CCK-8 assay and CCK-8 assay after digestion.The results were compared among different methods.Results After 5 d of culture,the cell spheroids were formed perfectly at the density of 2 375-10 000/well,and the volumes were in good linear with the original cell inoculation number at the density of 2 375-7 500/well.The A values of APH assay,MTT assay after digestion and CCK-8 assay after digestion increased with the increase of cell inoculation amount;But the cell ball digestion process was complex,and the cell viability was damaged.However,the A values of MTT and CCK-8 assay increased slowly.Conclusion The method of a hanging drop 3D cell culture model in 48-well culture plate combining with APH assay to detect cell viability is economical,accurate and easy to operate.
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Flavonol synthase (FLS) is one of the key enzymes in flavonoids metabolic pathways. In this study, middle sequence was obtained from Carthamus tinctorius transcriptome sequencing results. Full-length cDNAs of FLS was cloned from petals of C. tinctorius to FLS by using RT-PCR and RACE technology. Its full-length cDNA was 1,201 bp, with an open reading frame of 1,101 bp and 336 encoded amino acids. The phylogenetic analysis showed that, FLS gene encoded amino acids in C. tinctorius were highly homologous with amino acids in congeneric Compositae species, especially Rudbeckia laciniata. The pBASTA-FLS plant expression vector was successfully built by the molecular biology method, which lays a foundation for further studying biology functions of the gene and biosynthesis mechanism of flavonoids.