ABSTRACT
【Objective】 To establish a method to determine the content of the effective ingredient PCA (p-coumaric acid) in Shuang Bailian mixture and to investigate its anti-cancer mechanism. 【Methods】 High performance liquid chromatography (HPLC) was used to determine the content of PCA in Shuang Bailian mixture. The CCK8 method was used to detect the antitumor activity of PCA on esophageal cancer cells and the semi-inhibitory concentration of PCA on esophageal cancer cells. Moreover, the nude mice were used to investigate the anticancer effect of PCA. Western blotting was used to detect the expressions of apoptosis-related proteins (cleaved caspase 3, cleaved PARP, Bad, Bcl-2, PI3K, AKT, p-PI3K and p-AKT) in esophageal cancer cells and tumor tissues of nude mice. 【Results】 The concentration of PCA in Shuang Bailian mixture was 16.84 μg/mL. The linear regression equation of PCA was y=204 402x +360 904 (15-40 μg/mL), the RSD of the precision experiment was 2.66%, the RSD of the stability experiment was 2.35%, 3.22%, 1.58%, and 4.08%, respectively. The RSD of the repeatability experiment was 4.01%. The mean value of the recovery rate was 97.83% and the RSD value was 6.16%. CCK8 results showed that the half maximal inhibitory concentration (IC50) of PCA in KYSE30 and KYSE140 cells was 36 μg/mL and 55 μg/mL, respectively. The results of nude mice tumor experiments showed that after 30 days of administration, the tumor xenograft in control mice continued to increase in size, while the cisplatin group, PCA group, and PCA combined with cisplatin administration could effectively inhibit the growth of transplanted tumors in tumor-bearing mice. In addition, Western blotting results showed that compared with the control group, the cisplatin group, PCA group, and PCA combined with cisplatin group could effectively increase the protein expressions of cleaved caspase 3, cleaved PARP, and Bad, but decrease the protein expressions of Bcl-2, p-PI3K, and p-AKT in tumor tissues and KYSE30 and KYSE140 esophageal cancer cells. 【Conclusion】 The concentration of PCA in Shuang Bailian mixture was 16.84 μg/mL, and the HPLC content determination method conditions were sensitive and stable. PCA may have an active anti-tumor effect by regulating the PI3K/AKT/Bcl-2 signaling pathway and inducing apoptosis in esophageal cancer cells.
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Objective To examine the inhibition effect of zoledronic acid (ZOL) on malignant metastasis of human esophagus squamous cell carcinoma (ESCC) cells and to analyze its molecular mechanisms.Methods EC9706 and EC109 cells were treated with ZOL,and then MTT assay,adhesion and invasion assay were performed to observe the inhibitory effect of As2O3 on proliferation and metastasis of esophagus carcinoma cells.The expression of metastasis-related proteins was detected by Western blotting.Results Exposure to ZOL significantly presented suppressive functions on growth and metastasis of both kinds of cancer cells,in a dose-dependent manner(P＜ 0.05).Additionally,the expression level of occludin was increased after ZOL treatment by suppressing transcriptional factor Slug.Transfection of Slug could reverse anti-metastasis of ZOL.Conclusion ZOL possesses a significant anti-metastasis function on ESCC cells,mainly through repressing Slug to restore occludin expression.
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Objective To investigate the molecular mechanism of As 2 O3 in suppressing metastasis of esophagus carcinoma cells.Methods The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, adhesion and invasion assay were performed to observe the inhibitory effect of As 2 O3 on proliferation and metastasis of esophagus carcinoma cells .The expressions of matrix metalloproteinases ( MMP)2, MMP9, E-cadherin, and protein tyrosine phosphatase receptor-type O ( PTPRO) were analyzed with Western blot .Results Exposure to As 2 O3 significantly presented suppressive functions on growth and metastasis of esophagus carcinoma cells in a dose-dependent manner ( P <0.01 ) .Additionally , MMP2 and MMP9 expressions were increased after treatment with casticin ( P <0.01 ) , whereas E-cadherin and PTPRO expressions were down-regulated ( P <0.01 ) .Conclusions As2 O3 had a significant function to inhibit proliferation and metastasis of esophagus carcinoma cells .
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Objective To investigate the use of antibacterial drugs in tension -free inguinal hernia repair before and after the 2012 National clinical use of antibiotics special management ,providing the basis for the rational use of antimicrobial drugs and standardized management .Methods Retrospectively investigate the antimicrobial ap-plication in patients undergoing tension-free inguinal hernia repair and discharged from July to September in 2011-2013,and analyzed the timing of administration ,usage,type and treatment time of antimicrobial drugs .Results There were respectively 93.24%,47.76%and 27.59%of patients in the three groups administrated prophylaxis antibacte-rial drugs,and respectively 9.19%,65.67%and 85.08%of patients with indications .The first wound healing rates were respectively 94.59%,98.51% and 96.55%.The rates of reasonable choice of medicines 70.60%,96.88%and 91.67%,respectively;the rates of reasonable timing for medication were 71.01%,81.25%and 70.83%,respec-tively;the rates of reasonable courses of prophylaxis therapy were 33.33%,56.25% and 58.33% respectively. Conclusion The principle of no preventive antibiotics use in tension-free inguinal hernia repair is operable .After en-actment of special management of antibacterial drugs ,the level of preventive medication for tension-free inguinal herni-a repair is improved greatly .However ,it still needs to strengthen the management of antimicrobial prophylaxis timing and overall prophylaxis treatment course .
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[Objective] To purify and characterize a novel factor X activator,Fve-1 from Daboia russelli siamensis (Myanmar) venom.[ Methods]F V e-1 was purified by ion-exchange chromatography and gel filtration.The hemostatic activity of F V e-1 was determined based on chromogenic substrates.The fibrinogen-clotting activity of F V e-1 was also determined.Thermal stability, pH stability,enzyme activity,and inhibition of F V e- 1 were determined by its remaining procoagulant activity.N-treminal sequence was determined by the method of automated Edman degradation.[ Results ]F V e-1 was achieved by chromatography with a molecular weight of 13,808 and an isoelectric point of 4.6. The hemostatic activity of 0.5 mg Fve-1 was equal to that of 1.5625 u thrombin or that of 54.93 ng RVV X. F V e-1 primarily activated F X, but did not affect on prothrombin and fibrinogen. The suitable pH and temperature range of F V e-1 was 6.5-7.5 and 25-60 ℃,respectively.The activity of F V e-1 was enhanced by Ca2+ and inhibited by EDTA and DTT.The N-terminal sequence of F V e-1 was NH2-N-L-Y-Q-F-G-E-M-I-N.[Conclusion] F V e-1 is a factor X-activating enzyme,which could activate FX to FX a,but have minimal effect on prothrombin and fibrinogen.