ABSTRACT
Objective To observe the clinical effect of alprostadil combined with Yiqi Huoxue Huatan Tongluo Decoction in treating arteriosclerosis obliterans.Methods 60 cases of lower extremity arteriosclerosis occlusion disease outpatient and hospitalized patients were randomly divided into treatment group and control group according to the random number table method, each group were 30, the oral administration of enteric coated aspirin experimental group is set to control group intravenous injection of alprostadil treatment for 4 weeks, the treatment group was treated with Huatan Tongluo Decoction on the basis of the control group.The clinical symptoms, blood vessel function (ABI),low density lipoprotein-cholesterol (LDL-C), total cholesterol( TC) were observed in the two groups of patients, and blood rheology indexes included whole blood viscosity, fibrinogen, liver and kidney function and adverse reactions.ResuIts Total effective rate of the treatment group was 93.33% than control group 73.33% (P<0.05), the treatment group of ankle brachial index ( ABI) was better than the control group, the treatment group TC, LDL-C, high shear whole blood viscosity, low shear whole blood viscosity, fibrinogen protein improve were better than those of the control group (P<0.05).ConcIusion Disease curative effect is better than pure alprostadil in the treatment of occlusion Huoxue Huatan Tongluo Decoction combined with alprostadil in the treatment of lower limb arteriosclerosis.
ABSTRACT
Objective To detect the expression level of miRNA-575 by pancreatic cancer stem cells.Methods Pancreatic cancer cell lines were cultured with serum-free medium.The cancer stem cells were obtained and their characteristics were observed with an invasion assay,a xenograft experiment,and flow cytometry analysis.Moreover,qRT-PCR was used to detect the expression of miR-575.Results A small subpopulation of cells in both ASPC-1 and PANC-1 cells survived and formed spheres in the absence of serum.The mean number of migrated cells of PANC-1 and ASPC-1 spheres was (147.3±18.6) cells and (113.2±12.9) cells.Therefore,the spheres have higher invasive ability compared with those of parental cells.As few as 5 × 105 cells in PANC-1 and ASPC-1 spheres could initiate a xenograft tumor in NOD/SCID mice,which suggested the tumorigenicity of the spheres was 100 times those of parental cell lines.The percentages of CD24+CD44+ in spheres were 0.38％-0.43％ and 4.91％-5.21％,which showed increased expression compared with ASPC-1 and PANC-1 cell lines (P＜0.05).The expression of miR-575 in spheres was 3.71 and 3.83 times its expression in cell lines ASPC-1 and PANC-1 respectively (P＜0.05).Conclusions Pancreatic cancer cell spheres may be a source for collecting cancer stem cells in a serum-free medium culture,and MiR 575 may play an important role in regulating the biological characteristics of pancreatic cancer stem cells.
ABSTRACT
Objectives To isolate cancer stem cells (CSCs) in pancreatic cancer cell lines PANC1 and ASPC-1 with serum-free medium( SFM ), and to detect the expression of miR-590-3p in CSCs. Methods PANC1 and ASPC-1 cells was cultured in serum-free medium. The monoclonal formation, differentiation and cell cycle, half inhibitory concentration ( IC50 ), and the expression of the surface markers CD24 + , CD44 + were detected. qRT-PCR was used to detect the expression of miR-590-3p. Results After SFM culture, (0.94 ±0.53 ) ％ of ASPC-1 and (0.57 + 0. 12 ) ％ PANC1 survived, and they formed spheres, and could continuously passage in vitro. Cell spheres differentiation recurred when serum was supplemented in SFM. The G0/G1 stage proportion, CD24+ , CD44 + , CD24+ CD44+ cells proportion, IC50 in ASPC-1 cell were (75.3 ± 5.4)％,0.96％ ～ 2.01％, 27.52％ ～ 34.47％, 0.35％ ～ 0.44％ and (224.37 ± 5.71 ) μg/ml, which were significantly higher than that those in parent cell [ (43.7 ± 3.8 ) ％, 0. 38％ ～ 0.42％, 17.65％ ～ 18.25％,0.05％ ～0.08％, (11.43 ±2.10)μg/ml, P＜0.05]. The G0/G1 stage proportion, CD24+ ,CD44+ ,CD24 +CD44 + cells proportion, IC50 in PANC 1 cell were ( 80. 1 ± 4.7) ％, 5.31％ ～ 9.84％, 72.05％ ～ 93.06％,4.91％ ～5.21％, (296.58±4.27) μg/ml, which were significantly higher than that those in parent cell [ (46.1 ±5.3)％, 4.09％ ～4.97％, 47.71％ ～55.66％, 1.48％ ～2.63％, (26.17 ±3.81)μg/ml, P＜0.05]. The expression of miR-590-3p in ASPC-1, PANC1 spheres was 4.67 and 4.52 times higher than the expression in parent cell lines. Conclusions Pancreatic cancer cell spheres can be isolated from ASPC-1, PANC1 by culture with SFM. miR-590-3p is up-regulated and may play an important role in regulating biological characteristics of pancreatic cancer stem cells.