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Objective:To establish a method for the microbial limit test of Jingzhi Guanxin tablets. Methods: According to the methods in the 2010 edition and 2015 edition of Chinese Pharmacopoeia, the test was performed respectively. Results:Jingzhi Guanxin tablets showed obvious inhibitory effect on Bacillus subtilis. The bacteria count could be carried out by the culture medium diluting methods in the 2010 edition. The total amount of aerobe could be detected by the membrane filtration method in the 2015 edition. The total combined molds and yeasts count could be performed by the plate count method and the specified microorganism could be tested with the routine method. Conclusion:The above methods can be used for the microbial limit test of Jingzhi Guanxin tablets.
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Objective To investigate the effects of chitosan(CS)/pcDNA3.1(+) CrmA on the expression of interleukin-13 (IL-1β3) converting enzyme (ICE) and IL-1β in chondrocytes.Methods Rabbit chondrocytes were isolated and cultured.Chondrocytes were treated with PBS,10 μg/ml CS/pcDNA3.1(+) and CS/pcDNA3.1 (+) CrmA respectively for 6 hours.Then 10 ng/ml IL-1β was added into the culture medium.After 48 hours,the messenger RNA and protein expression of ICE and IL-1β in chondrocytes were detected by using real time polymerase chain reaction and western blotting.Results In CS/pcDNA3.1 (+) CrmA treated group (0.52 ±0.09),the mRNA expression of ICE inchondrocytes was significantly inhibited compared withcorresponding samples of CS/pcDNA3.1 (+) group (0.84±0.11,t=4.42,P＜0.01) and PBS group (1.00±0.10,t=6.58,P＜0.01).ICE protein expression in CS/pcDNA3.1 (+) CrmA treated group (0.20±0.03) was markedly lower than CS/pcDNA3.1 (+) (0.37±0.05,t=4.85,P＜0.01) and PBS treated group (0.44±0.07,t=6.68,P＜0.01).There was no significant difference of ICE mRNA and protein expressions in chondrocytes between CS/pcDNA3.1(+) group and PBS group.Significant difference of IL-1β mRNA expression was found in the three groups.IL-1 β mRNA expression level was significantly lower in CS/pcDNA3.1(+) CrmA treated group (0.55± 0.08) than CS/pcDNA3.1(+) (0.69±0.06,t=3.50,P＜0.01) and PBS group (0.99±0.04,t=11.12,P＜0.01) of chondrocytes.IL-1β protein expression level of chondrocytesin CS/pcDNA3.1(+) CrmA group (0.230±0.020) was significantly lower than CS/pcDNA3.1 (+) (0.450±0.060,t=5.07,P＜0.01)and PBS groups (0.610±0.090,t=8.70,P＜0.01) of cells.Significant difference of IL-1β mRNA and protein expression between CS/pcDNA3.1 (+) and PBS group was also observed (t=7.61,P＜0.01;t=3.63,P＜0.01).Conclusion CrmA mediated by chitosan can significantly suppress the mRNA and protein expression of ICE,thus down regulat the expression of IL-1β,which may be one of the mechanisms of CS/pcDNA3.1 (+) CrmA in the treatment of osteoarthritic cartilage degeneration.
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Objective:To study the application of petrifilm colony count plate in the microbial detection for cosmetics. Methods:Three common microbials in 14 batches of cosmetics were respectively detected by the method described in hygienic standard for cos-metics and petrifilm colony count plate, and the results were compared. Results:The results shown by the two methods had no statisti-cally significant difference (P>0. 05). Conclusion: Petrifilm colony count plate with light weight, fast and easy operation exhibits significance in the microbial detection for cosmetics.
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Objective:To establish a Taqman fluorescence quantitative PCR for the detection of Salmonella in drugs. Methods:Based on Salmonella specific gene, a pair of primers and a probe were designed, and DNA of Salmonella was extracted for the detec-tion. Results:The method was much specific, and the detection limit was 160 cfu/ml. The detection rate was 100% for the artificially contaminated samples. Conclusion:Fluorescence quantitative PCR can be applied in the rapid detection of Salmonella in drugs.
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Objective To investigate the effects of chitosan/pCDNA3.1 (+) CrmA on matrix metalloproteinase (MMP)-1 and tissue inhibitor of metalloproteinase (TIMP)-1 expression of chondrocytes induced by interleukin-1β (IL-1β) in mRNA and protein levels.Methods Rabbit chondrocytes were isolated and cultured.Chondrocytes were treated with PBS,10 μg/ml chitosan (CS) and chitosan/ pCDNA3.1 (+)CrmA respectively for 6 hours.Then 10 ng/ml IL-1β was added into the culture medium.After 48 hours,real time polymerase chain reaction (real time PCR) and Western blot assay were used to examine the changes of MMP1 and TIMP-1 in mRNA and protein levels.Results In CS/pCDNA3.1 (+)CrmA treated group (0.44±0.04),the messenger RNA expression of MMP-1 in chondrocytes was significantly suppressed compared with corresponding samples of PBS treated group (1.00±0.05) and CS treated group (0.76±0.07),There was significant difference between three groups (F=106.93,P＜0.01).The MMP-1 protein expression of chondrocytes in CS/pCDNA3.1 (+)CrmA treated group (0.28±0.03) was lower than that of PBS treated group (0.73±0.06) and CS treated group (0.46±0.05)(F=59.66,P＜0.01).No significant difference of TIMP-1 expression among the three groups was observed in mRNA and protein levels.Conclusion CS/pCDNA3.1(+) CrmA can significantly inhibit the mRNA and protein expressions of MMP-1 of chondrocytes induced by IL-1β,which leads to up-regulation the ratio of TIMPs to MMPs of IL-1β induced chondrocytes.It may be a part of the mechanisms of the therapeutic effects of CS/pCDNA3.1 (+)CrmA on osteoarthritis.