AIM:To explore the effect and mecha-nism of ethanol extract of Angelica sinensis(Oliv.)Diels(EEA)on cell proliferation and apoptosis in B16-F10 melanoma cells.METHODS:Cell viability was analyzed by MTT method.Cell proliferation was detected by colony formation assay.The invert-ed microscope was used to observe the changes of cell growth confluence and morphology.Hoechst 33342 staining was used to detect cell apoptosis.Flow cytometry(FCM)was used to detect cell cycle and apoptosis.Transmission electron microscopy(TEM)was used to observe the changes of cell mi-tochondrial structure.Western blot was used to de-tect the expression levels of cell cycle,apoptosis,mitochondrial biogenesis,and mitochondrial dy-namics-related proteins.RESULTS:Compared with the blank control group,the cell viability of B16-F10 melanoma cells was reduced after EEA(10-400μg/mL)treatment for 24 h and 48 h,respectively(P＜0.05,P＜0.01).The decreased cell growth conflu-ence,morphological changes such as shrinkage,rounding,and reduction in the volume,and apop-totic morphologic changes such as chromatin con-densation were observed after EEA(100 μg/mL and 200 μg/mL)treatment for 24 h.The number of cell clones was decreased after EEA(10-200 μg/mL)treatment for 14 d(P＜0.01).The morphology of mitochondria became more round and shorter,and the inner mitochondrial matrices were either damaged or absent after 200 μg/mL EEA treatment for 24 h.The ratio of cells in G0/G1 phase and the early apoptosis rate of cells were higher than those of the blank control group(P＜0.01)after EEA(20-200 μg/mL)treatment for 24 h.Western blot re-sults showed that compared with the blank control group,the protein expression levels of cleaved cas-pase-9,Bax,DRP1,and FIS1 were up-regulated(P＜0.05,P＜0.01),and the protein expression levels of cyclin D1,cyclin E,CDK2,CDK4,Bcl-2,Bad,Bcl-XL,SIRT1,PGC-1α,NRF1,TFAM,MFN2,and OPA1 were down-regulated(P＜0.05,P＜0.01).CONCLUSION:EEA has an inhibitory effect on the proliferation of B16-F10 melanoma cells,which may be related to the induction of G1/S cell cycle arrest and mito-chondrial apoptotic pathway,and the disruption of mitochondrial biogenesis and mitochondrial dy-namics.

AIM:To explore the effect and mecha-nism of ethanol extract of Angelica sinensis(Oliv.)Diels(EEA)on cell proliferation and apoptosis in B16-F10 melanoma cells.METHODS:Cell viability was analyzed by MTT method.Cell proliferation was detected by colony formation assay.The invert-ed microscope was used to observe the changes of cell growth confluence and morphology.Hoechst 33342 staining was used to detect cell apoptosis.Flow cytometry(FCM)was used to detect cell cycle and apoptosis.Transmission electron microscopy(TEM)was used to observe the changes of cell mi-tochondrial structure.Western blot was used to de-tect the expression levels of cell cycle,apoptosis,mitochondrial biogenesis,and mitochondrial dy-namics-related proteins.RESULTS:Compared with the blank control group,the cell viability of B16-F10 melanoma cells was reduced after EEA(10-400μg/mL)treatment for 24 h and 48 h,respectively(P＜0.05,P＜0.01).The decreased cell growth conflu-ence,morphological changes such as shrinkage,rounding,and reduction in the volume,and apop-totic morphologic changes such as chromatin con-densation were observed after EEA(100 μg/mL and 200 μg/mL)treatment for 24 h.The number of cell clones was decreased after EEA(10-200 μg/mL)treatment for 14 d(P＜0.01).The morphology of mitochondria became more round and shorter,and the inner mitochondrial matrices were either damaged or absent after 200 μg/mL EEA treatment for 24 h.The ratio of cells in G0/G1 phase and the early apoptosis rate of cells were higher than those of the blank control group(P＜0.01)after EEA(20-200 μg/mL)treatment for 24 h.Western blot re-sults showed that compared with the blank control group,the protein expression levels of cleaved cas-pase-9,Bax,DRP1,and FIS1 were up-regulated(P＜0.05,P＜0.01),and the protein expression levels of cyclin D1,cyclin E,CDK2,CDK4,Bcl-2,Bad,Bcl-XL,SIRT1,PGC-1α,NRF1,TFAM,MFN2,and OPA1 were down-regulated(P＜0.05,P＜0.01).CONCLUSION:EEA has an inhibitory effect on the proliferation of B16-F10 melanoma cells,which may be related to the induction of G1/S cell cycle arrest and mito-chondrial apoptotic pathway,and the disruption of mitochondrial biogenesis and mitochondrial dy-namics.

Objective: To evaluate the efficacy and safety of Oryz-Aspergillus enzyme and pancreatin tablets (Combizym^®) in the treatment of postprandial distress syndrome (PDS) in the elderly, compared with gastrointestinal motility drugs. Methods: A prospective randomized controlled trial was designed and registered in the China Clinical Trials Registry (ChiCTR-IPR-16008185). The elderly patients with PDS were randomly divided into three groups, including Mosapride group with Mosapride citrate tablets 5 mg 3 times per day for 2 weeks; Combizym^® group with Combizym tablets 244 mg 3 times per day for 2 weeks; combined treatment group with both drugs and same doses for 2 weeks. The modified Nepean dyspepsia index (NDSI) score, discomfort intensity score and PDS score were calculated on patients before treatment, at the end of first and second week of treatment, as well as 4 weeks after treatment finished, respectively. Adverse effects were evaluated. Results: A total of 323 patients from 16 tertiary hospitals in China were enrolled in this study. Among them, 105 patients were in Mosapride group, 109 in Combizym^® group and 109 in combined treatment group. There were 148 males (45.8%) and 175 females (54.2%) with median age 71.4±9.0 years (60-100 years). Baseline characteristics of three groups were comparable. After treatment, the NDSI scores in three groups all decreased significantly (P<0.001), while they were similar between groups (P>0.05). The discomfort intensity score and PDS score in three groups showed a significant reduction after treatment (P<0.001), especially in the combined treatment group. Compared with Mosapride group, the scores in Combizym^® group decreased significantly after one or two weeks [discomfort intensity score: after one week, 4.0(2.5, 8.0) vs. 6.0(3.0, 10.0); after two weeks, 3.0(0.0, 5.0) vs. 4.0(2.0, 6.0); all P<0.05. PDS score: after one week, 6.0(3.0, 9.0) vs. 7.0(3.5, 10.5); after two weeks, 3.0(0.0, 5.0) vs. 4.0(2.0, 7.0); all P<0.05]. The efficacy rate in all patients after first week of treatment was over 15.0%. The efficacy rates after two weeks were 55.2%, 68.8% and 73.4% in Mosapride group, Combizym^® group and combined treatment group, respectively. After two week treatment, the efficacy rates in Combizym^® group (P=0.041) and combined group (P=0.006) were higher than that of Mosapride group. The recurrence rate of Mosapride group was 9.5%, which was significantly higher than that of Combizym^® group (1.8%, P<0.05) and combined treatment group (1.8%, P<0.05). There were no serious adverse effects in the three groups. Conclusions The efficacy of Oryz-Aspergillus enzyme and pancreatin tablets is comparable with that of Mosapride in elderly PDS patients, with fewer adverse effects and low recurrence rate. Combination regimen indicates better efficacy than that of Oryz-Aspergillus enzyme and pancreatin tablets or Mosapride alone.

Objective To investigate the association of serum lipid,lipoprotein,apolipoprotein,leptin,cholecystokinin(CCK)and bile lipid with cholesterol gallstone formation.Methods The patients with gallstone were divided into cholesterol(n=99)and non-cholesterol(n=57)gallstone groups by infrared spectometry.And 52 healthy volunteers were served as control group.The concentrations of serum cholesterol,triglyceride,high and low density lipoproteins,apolipoprotein(Apo),leptin and CCK were measured and compared among three groups.The levels of total bile cholesterol,bile acid and lecithin were also detected.Results The concentrations of serum triglyeeride and total cholesterol in two gallstone groups were higher than those in control group(P value all＜0.01).The level of Apo-B in cholesterol gallstone group was higher than that in control group(P=0.017).While the concentrations of high density lipoprotein and CCK were significantly lower in two gallstone groups than those in control group(P value all=0.000).Serum leptin was higher in male patients compared to controls(P＜0.05).The bile cholesterol saturation index in two gallstone groups was above 1.Conclusions The changes of serum CCK,triglyceride,total cholesterol,high density lipoprotein,Apo-B and leptin may be correlated to the formation of gallbladder gallstone.