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This study aimed at the molecular mechanism of Cordyceps sinensis polysaccharide-A(CPS-A) on angiotensin (Ang Ⅱ)-induced injury of L02 cells.The effect of AngⅡ and CPS-A on the proliferation of L02 cells was analyzed by MTT assay.PCR,Real-Time PCR and Western blot were also employed to determine the expression of IL-1β,AT1R,AT2R,NF-κB p65,TNFα and other inflammatory factors at mRNA and protein levels.The results showed that Ang Ⅱ and CPS-A could inhibit the proliferation of L02 cells by 1 × 10-5 mol/L and 200 μg/ mL,respectively.PCR,Real-Time PCR and Western blot showed that CPS-A could significantly down-regulate IL-1 β,TNF-α,NF-κB and AT1R.CPS-A has a good protective effect on AngⅡ-induced L02 cell injury.
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Objective:To construct a chemiluminescense immune quantification assay based one paired mAbs against complexed prostate specific antigen ( c-PSA).Methods:Six week-old female BALB/c mice were immunized with the commercial c-PSA antigen.After serum titer reaching a platform stage ,the spleen was immunized and fused with mouse myeloma cell lines ( Sp2/0 ) .The hybridoma were screened by indirect ELISA ,and eight generated antibodies were paired to obtain a quantitative analysis of the chemical luminescence.Results:7D6 specifically recognized c-PSA,while 1A10 recognized total PSA(t-PSA).And the paired antibody 1A10/7D6 were determined to successfully construct a chemiluminescense immune response quantitative detection method through the detection of c-PSA standard and clinical serum samples .had,positive samples have statistically significant difference ( P<0.000 1 ) with negative samples.And the correlation coefficient R 2 was 0.97 between our c-PSA quantitative results and that of the Siemens c-PSA chemiluminescense immunoassay kit .The detection linear range was 0.1-100 ng/ml,and the sensitivity was 0.005 ng/ml.Conclusion:The paired monoclonal antibodies specifically detecting c-PSA were generated and a c-PSA chemiluminescense immunoassay were developed in this study .The detection capability of our method was comparable with that of the international commercial kit .
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Objective To evaluate the application of partial cpsA-cpsB serotype prediction system as a serotyping method for streptococcus pneumonia.Methods Ninety-four isolates in this study were provided by Microorganism Research Room of Beijing Pediatric Research Institution,Beijing Children's Hospital Affiliated to Capital Medical University.The quelling test was applied to determine gold standard of serotypes of isolates.Polymerase chain reaction (PCR),sequencing,sequence data management and alignment were implemented previously.Results Eighty-three out of all 94 isolates were serotyped by quelling reaction,and 11 isolates were non-serotype isolates.Among the 83 isolates,67 (80.72％) isolates got positive PCR results and 60 (89.55％)isolates got results consistent with gold standard or containing gold standard.Among 12 isolates belonging to 19F,10 isolates were correctly predicted,and 2 isolates were predicted to be 6A,23F/10A.Among 19 isolates belonging to serotype 19A,1 isolate was predicted to be 35 F/47F,and the other 18 isolates were correctly predicted.Among 10 isolates belonging to serotype 14,9 isolates got results consistent with gold standard,and 1 isolate was predicted to be 19A.All 7 isolates belonging to serotype 6B were predicted to be 6A/6B and 4 isolates belonging to 23F were predicted to be 23F/10A.3 of 11 (27.27％) non-serotype isolates got positive PCR results and were predicted to be 6A/6C,6A/6B,19A.Conclusions Partial cpsA-cpsB sequencing system is a useful method for detecting streptococcus pneumoniae serotypes.