ABSTRACT
Objective To investigate the effects of silencing STMN1 by siRNA on the sensitivity of oesophageal cancer cells Eca-109 to paclitaxel .Methods The STMN1 siRNA(siSTMN1) or scramble siRNA(SCR) were transient transfected into Eca-109 cells .The mRNA and protein levels of STMN1 were detected by qPCR and Western blot in the Eca-109 cells of different groups .In vitro paclitaxel sensitivity of siSTMN1 and SCR transfected Eca-109 cell lines was tested by MTT assay and colony formation as-say .Hoechst 33258 nuclear staining were used to investigate the effect of silencing STMN 1 on the sensitivity of SCR ,siSTMN1 transfected Eca-109 cells and nontreated counterparts under paclitaxel induced apoptosis .Results The transient transfection cell lines were successfully established .Both protein and mRNA levels of STMN1 were effectively down-regulated in the siSTMN1 transfected Eca-109 cells .Down-regulation of STMN1 significantly enhanced the sensitivity of Eca-109 cells in response to paclitaxel (P<0 .01) .In addition ,the siSTMN1 transfected Eca-109 cells displayed significant apoptosis as assessed by Hoechst nuclear stai-ning(P<0 .01) .Conclusion Silencing STMN1 by siRNA could enhance the sensitivity of oesophageal cancer cells Eca-109 to paclitaxel .
ABSTRACT
Objective To evaluate the changes to mitochondrial ultrastructure in melanocytes of perilesional skin from patients with vitiligo.Methods Skin specimens were obtained from the perilesional area (0.5-1 cm distal to vitiligo lesions) of 10 patients with progressive vitiligo and 10 patients with stable vitiligo,as well as from the normal skin of 10 healthy volunteers.The morphology of melanocytes was observed by using transmission electron microscopy (TEM).Besides,stereological parameters of mitochondria,such as volume density (Vv),surface density (Sv) and numerical density (Nv),were measured.Results In melanocytes from the healthy controls,there were a large number of melanosomes with the number of melanosomes per melanocyte being 28.57± 3.21,which were mainly at stage Ⅲ and Ⅳ; mitochondria with normal structure and densely packed cristae were regularly arranged; autophagosomes were seen occasionally.Compared with the melanocytes from healthy controls,there was an obvious decrease in the number of melanosomes (especially stage Ⅲ melanosomes) in melanocytes from the perilesional skin of patients,with the number of melanosomes per melanocyte being 22.00 ± 6.16 (P ＜ 0.05) and 17.43 ± 6.24 (P ＜ 0.05) in patients with progressive vitiligo and stable vitiligo,respectively.TEM also showed disorganized or disrupted mitochondria in various shapes and sizes,most of which were swelling with obscure cristae and vacuolization,in melanocytes from the perilesional skin,and no autophagy was observed.The three stereological parameters were significantly different between the three groups of tissue specimens (all P ＜ 0.05),with the Nv,Vv and Sv of mitochondria being (7.194 ± 1.434) μm-3,(4.8 ± 1.2) ％,(2.42 ± 0.86) μ m-1 respectively in melanocytes from the healthy controls,(4.055 ± 0.906) μm-3,(7.4 ± 2.1)％,(3.58 ± 1.15)μm-1 respectively from patients with progressive vitiligo,(5.311 ± 0.873) μm-3,(6.5 ± 1.4)％,(2.82 ± 0.94) μm-1 respectively from patients with stable vitiligo.Conclusions Mitochondria are injured in melanocytes from perilesional skin of patients with vitiligo,and the degree of injury is more intense in progressive vitiligo than in stable vitiligo.
ABSTRACT
Objective To measure the expression of granulocyte colony-stimulating factor receptor (G-CSFR) in human melanocytes and to evaluate the biologic effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on human melanocytes.Methods Melanocytes were obtained from circumcision specimens of healthy males,and neutrophils were isolated from heparin-andcoagulated peripheral blood of healthy human followed by a primary culture.Then,the melanocytes in third passage were cultured with or without the presence of various concentrations (200,400,600,800 μg/L) of rhG-CSF for 72 hours.The growth and morphology of melanocytes were observed.Flow cytometry was performed to detect the expression of G-CSFR in untreated human melanocytes,neutrophils and erythroleukemia cells (HEL 92.1.7).Western blot and reverse transcription PCR (RT-PCR) were carried out to measure the expression of G-CSFR protein and mRNA respectively in the neutrophils,HEL 92.1.7 cells,treated or untreated human melanocytes.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferation,and dopa-oxidation assay to estimate the tyrosinase activity,of treated melanocytes.Results The expression rate of G-CSFR was 76.81％ ± 10.70％ in human melanocytes,significantly higher than that in the HEL 92.1.7 cells (2.53％ ± 1.54％,P ＜ 0.01 ),but lower than that in the neutrophils (85.76％ ± 15.71％,P ＜ 0.05).Both G-CSFR protein and mRNA were expressed in melanocytes,and there was no significant differences in the expression level of G-CSFR protein and mRNA among melanocytes treated with different concentrations of rhG-CSF (both P ＞ 0.05).The expression levels of G-CSFR protein and mRNA in the melanecytes were significantly higher than those in the HEL 92.1.7 cells (both P ＜ 0.01 ),but lower than those in the neutrophils (P ＜ 0.05 or ＜ 0.01 ).rhG-CSF at 200-800 μg/L displayed a significantly promotive effect on the proliferation of melanocytes (P ＜ 0.01 or ＜ 0.05 ),and the effect was in a dose-dependent manner when rhG-CSF ranged from 200 to 600 μg/L (P ＜ 0.01 ).The rhG-CSF at 600 μg/L and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) at 20 μg/L showed an equivalent effect on the proliferation of melanocytes (164.04％ ± 13.0％ vs.165.62％ ± 10.6％,P ＞ 0.05).However,rhG-CSF from 200 to 800 μg/L had no significant impact on the tyrosinase activity of melanocytes (all P ＞ 0.05 ).Conclusions G-CSFR is expressed in human melanocytes. rhG-CSF can promote the proliferation of cultured human melanocytes,but has no obvious influence on the tyrosinase activity of melanocytes.
ABSTRACT
Objective To evaluate the clinical effects of early endoscopic therapy for elderly patients with acute severe biliary pancreatitis.Methods Ninety-two elderly patients with acute severe biliary pancreatitis were randomly divided into 2 groups:ERCP group ( n =43) and non-ERCP group ( n =49).Serum TNF-a,IL-6,IL-8,the recovery time of blood amylase,the duration of abdominal pain,hospitalization,mortality and complications were compared.Results ERCP group showed a greater decrease in serum TNF-α,IL-6 and IL-8 levels than the control group ( 45.16 ± 13.48 ) μg/L v.s.( 176.89 ± 47.35 ) μg/L,(31.76 ± 13.85)μg/L v.s.(68.48 ±24.87) μg/L,(113.39 ±63.78) μg/L v.s.(309.86 ± 117.13)μg/L) (P ＜0.05 ).The duration of abdominal pain,the recovery time of blood amylase and hospitalization in ERCP group were significantly shorter compared to the non-ERCP group [ ( 10.2 ± 1.7 ) d v.s.( 13.2 ±2.4)d,(3.3 ±1.0)dv.s.(5.5 ±1.2)d,(15 ±1.6)dv.s.(20±3.0)d] (P＜0.05),and complication rate of the ERCP group was lower,too (5％ v.s.22％,P ＜ 0.05).Conclusion Early ERCP is safe and highly effective for the elderly patients with acute severe biliary pancreatitis.
ABSTRACT
Objective To evaluate effectiveness of prevention and treatment for patients with chronic obstructive pulmonary disease (COPD) for one year at community health-care service (CHS)centers. Methods A quasi-experiment design was used to evaluate effectiveness of community management for patients with COPD who visited and registered at Xiaokunshan and Xinbang CHS centers in Songjiang district of Shanghai in 2008, with COPD patients from Xiaokunshan community CHS center as management group and those from Xinbang as control. Measures for community management included training for local general practitioners in essential knowledge of COPD prevention and treatment, allocation of necessary drugs for COPD treatment, and health education for COPD patients and their family members. No special measures were taken for control group except routine treatment. Results A total of 132 patients were diagnosed as COPD according to their pulmonary function on 2008, 61 at Xiaokunshan and 71 at Xinbang CHS canters.One hundred and two patients, 47 at Xiaokunshan and 55 at Xinbang, finished one-year follow-up. Five patients from Xiaokunshan who were smokers at their first visits and three of them quitted smoking after oneyear management. Meanwhile, none of nine smoked patients from Xinbang quitted smoking in the same period. There was no statistically significant difference in symptom scores, pulmonary function, and forced expiratory volume at the first second (FEV1) between patients in the two groups during one-year follow-up.Average score of quality of life in patients at Xiaokunshan dropped to 39. 12 after one-year management from 46. 96 at their first visits, and that in patients at Xinbang increased to 62. 11 from 56. 55 (P＜0.01).Average six-minute walking distance (6-MWD) in patients at Xiaokunshan reduced to 354. 26 meters after one-year management from 361.66 meters at their first visits, meanwhile that in patients at Xinbang reduced to 351.18 meters from 398.07 meters (P =0. 008). Scores of functional dyspnea in patients at Xiaokunshan fell to 0.34 from 0.40 at their first visit, meanwhile that in patients at Xinbang increased to 1.00 from 0.95(P =0.038). During the one-year follow-up, 13 patients at Xiaokunshan and 53 at Xinbang visited emergency departments for treatment, and four at Xiaokunshan were hospitalized and none died, and five at Xinbang were hospitalized due to acute exacerbation of COPD and four of them died. Conclusions The study suggests that strengthening prevention and treatment for COPD patients at community level by general practitioners, patients themselves and their family members can improve their quality of life, reduce emergency visits and hospitalization, alleviate dyspnea symptoms and delay the decline of 6MWD.
ABSTRACT
Objective To investigate the regulation of metabolize characteristic of some antibiotics in ACST in human bile, and provide the theoretic basis for physician selecting antibiotic rationally when biliary tract has been infected. Methods Samples of ACST in human bile were obtained by ERCP + ENBD, contents of Cefradine, Cefoperazone, Ceftriaxone, Ciprofloxacin and Lomefloxacin in human bile were simultaneously measured by HPLC method. Results After antibiotics were given through iv 0. 25 ～ 1. 5 hour, an increase was observed in contents of all antibiotics in bile, and the changes of Ceftriaxone and Ciprofloxa-cin were the most significant ( P <0.01). Ceftriaxone and Ciprofloxacin had a higher Cmax than other antibiotics, and Cefoperazone had a longer tl/2. Conclusion As maintaining a little time to tmax , a higher Cmax and a longer tl/2 in the bile by iv, Cefradine and Ciprofloxacin are recommended in the patient with ACST.
ABSTRACT
uate and should be standardized.
ABSTRACT
Objectives:To observe growth suppressive effects of human wild type p53(wt p53) gene on human esophageal cancer cell line. Methods:Using the retroviral vector to introduce exogenous wt p53 gene into human esophageal cancer cell line ECA109,the gene expression and tumor inhibition were studied in vitro and in vivo . Results:The expression of p53 in transfected cell lines(ECA 109/p53 ) was increased.The growth rates and the ability to form colony in soft agar were greatly inhibited in ECA 109/p53 cells versus ECA109/neo and ECA109 cells.The G 0+G 1 ratio increased and S ratio decreased in cell cycle distribution,and apoptosis index significantly rose in the ECA109/p53 cells,which were confirmed by FCM analyzing.The tumorigen icity of ECA109/p53 cells in nude mice was obviously suppressed by exogenous p53. Conclusions:Exogenous wt p53 mediated by retroviral vector could inhibit the growth of human esophageal cancer cell.
ABSTRACT
AIM To study the pharmacokinetics and bioavailability of clinafloxacin in rats. METHODS The drug concentration was determined by HPLC. The main pharmacokinetic parameters were obtained by 3P87 program. An RP-C18 was used as the stationary phase. The mobile phase was a mixture of acetonitrile-0.05 mol*L-1 citric acid triethylamine (pH 2.5) (20∶80). The flow rate was 1.0 mL*min-1. The UV absorbance detector was set at 300 nm. RESULTS A good linearity was obtained from 0.03-20 μg*mL-1 of clinafloxacin in rat plasma with γ=0.9998. The plasma concentration-time curve of clinafloxacin conformed to one compartment open model. After ig administration of 50 mg*kg-1 and 100 mg*kg-1 dose of clinafloxacin in six rats, mean Cmax and AUC values increased in proportion to dose. Mean T1/2 appeared to be independent of dose. Mean AUC was 65±6 and 27±4 μg*h*mL-1 respectively after iv and ig adminostration of 100 mg*kg-1 dose. The extent of bioavailability (F) of clinafloxacin was 42%. CONCLUSION The results of the pharmacokinetic study of clinafloxacin showed that it exhibited first order kinetic characteristics and the bioavailability is low.