1.Evaluation of different methods in eliminating Mycoplasma contamination in cell culture of human hepatoma-derived cell line C3A
Yang LI ; Chaoyi FU ; Jun WENG ; Yi GAO ; Qing PENG
Chinese Journal of Microbiology and Immunology 2016;36(10):766-770
Objective To evaluate the effectiveness of several methods in eliminating Mycoplasma contamination in cell culture of human hepatoma-derived cell line C3A. Methods PCR was performed to detect the Mycoplasmas contamination in cell cultures. The contaminated samples were treated by ciprofloxa-cin, heating, Plasmocure or co-culturing with macrophages. Transmission electron microscope ( TEM) and Q-PCR were used to comparatively analyze the cell morphology and gene expression before and after Plas-mocure treatment. Results Plasmocure succeeded in eliminating Mycoplasma contamination, while cipro-floxacin showed temporary efficacy. Heating and co-culturing with macrophages failed to eliminate Mycoplas-ma contamination. No Mycoplasma contamination in the Plasmocure-treated group was observed under TEM and the expression of ALB, TF and CYP3A4 genes were higher than the genes expressed in the contaminated group (P<0. 01). Conclusion Plasmocure treatment was effective in eliminating Mycoplasma contamina-tion in cell culture. Moreover, the cell morphology and gene expression in Plasmocure-treated group were re-stored to normal.
2.Prescription compatibility effect on the major components absorption in danshen extract and their identification.
Li LIN ; Jianxun LIU ; Ying ZHANG ; Chaoyi MAO ; Changling DUAN ; Jianhua FU ; Chengren LIN
Acta Pharmaceutica Sinica 2010;45(10):1290-5
An improved everted gut sac method was applied to the study of prescription compatibility effect on the major components in Danshen extracts. With the separation and detection by HPLC-ECD, 5 major peaks could be detected in intestinal absorbed solution after prescription administration. Following the identification by HPLC-MS/MS, peak 2, 3, 4, and 5 were rosmaric acid, lithospermic acid, salvianolic acid B, and salvianolic acid A, respectively, which also confirmed with reference standards of those components. Through paralleling substance identification, peak 2, 3, 4, and 5 could be found as the major components in Danshen extracts, except Salvianolic acid E which is undetectable in intestinal solution. The contents of peak 2, 3, and 4 did not show difference before and after compatible prescription administrated, where the peak 5 had a significant increase in the same process. Those results revealed that peak 5, salvianolic acid A, might lead to an increasing pharmacological effect after prescription compatibility.
3.Experimental study of Tibet miniature pig model of acute renal failure treated by a novel hybrid liver and renal support system
Lei FENG ; Guolin HE ; Longhui XIONG ; Chaoyi FU ; Xiaolin HUO ; Yi GAO
Chongqing Medicine 2016;45(8):1009-1011,1015
Objective To evaluate the novel hybrid liver and renal support system the safety and efficacy of the treatment of Tibet mini pig model of renal failure .Methods Five Tibet miniature pig bilateral renal artery ligation approached to construct the models of renal failure ,treatment group was treated with therapy (CRRT mode) of the novel hybrid liver and renal support system after modeling ,while the control group received no treatment .Drawning venous blood endotoxin ,biochemical tests and aerobic anae‐robic culture when in the treatment of 0 ,4 ,8 h;then we observed the changes of biochemical indicators in Tibet mini pigs before and after treatment ;we recorded machine pressure monitoring indicators every 2 hour during treatment .Results The model of renal failure of two experimental animals were successfully constructed .The general condition of treatment group was improved after treated by the novel hybrid liver and renal support system ,while the control groups continued to deteriorate .The experimental ani‐mals did not appear abnormal ECG during the experiment .Two groups of animals showed no statistics significant difference (P>0 .05) in the period of 0 h .In the remaining period ,heart rate and respiratory rate in treatment group was significantly reduced(P<0 .05) ,while oxygen saturation and mean arterial pressure were significantly increased compared to control group(P<0 .05) .There was no differences of biochemical indicators between two groups before modeling (P>0 .05);after modeling both groups biochemi‐cal indicators was significantly increased compared with baseline (P<0 .05);after treatment of the hybrid liver and renal support systems in the treatment group ,the biochemical parameters were significantly decreased compared with baseline ,the difference was statistically significant different(P<0 .05) ,while the control group did not change significantly .Blood endotoxin measured results were less than 0 .5 EU/mL ,arteriovenous end aerobic anaerobic culture showed no bacterial growth .Pathological examination showed significant swelling of the renal tubular epithelium and tubular necrosis .Conclusion The novel hybrid liver and renal sup‐port system for the treatment of Tibet mini‐pig model of renal failure is safe and effective ,and it can be further attempts for clinical trials .