AIM: To explore the synthesis of thermo-sensitive poly N-isopropylacry-lamide(PNIPAAm)and the petri dish grafted with PNIPAAm hydrogels by the electron accelerator, as well as the growth conditions and the biological characteristics of rabbit corneal stromal cells on thermo-sensitive PNIPAAm hydrogels, and the cell sheets obtained from the PNIPAAm hydrogels.METHODS: NIPAAm monomer was dissolved in 2-propanol at concentrations of 55% with 0.5% N,N'-Methylenebisacry-lamide(MBA). Solution(70 μL)was added and spread uniformly over 35 mm petri dish. These dishes were immediately subjected to irradiation. After follow-up treatment, rabbit corneal stromal cells were cultured on thermo-sensitive petri dish in vitro.RESULTS: According to the monomer formula and radiation synthesis scheme in this experiment, PNIPAAm can be synthesized on the surface of the petri dish. Rabbit corneal stromal cells grew well in the thermo- sensitive surface and can be separated into sheets.CONCLUSION: The single and multilayer carrier-free cell sheets can be obtained from the use of thermo-sensitive petri dish.

ObjectiveTo investigate the efficacy of intensity-modulated radiotherapy (IMRT) in treating large primary hepatocellular carcinoma (LHCC) which is unsuitable for surgery or has poor response to radiofrequency ablation, interventional therapy, and other local treatments, and to identify the prognostic factors for survival. MethodsWe retrospectively analyzed the clinical data of 29 LHCC patients who received IMRT from April 2008 to August 2011. There were five fractions per week and the dose for each fraction was 2 to 6 Gy; the total dose was 50 to 70 Gy. The short-term efficacy and prognosis were observed and analyzed. The Kaplan-Meier method was used to calculate survival rates and the log-rank test was used for survival difference analysis. Multivariate analysis was performed using the Cox regression model. ResultsThe complete remission, partial remission, stable disease, and disease progression rates were 3.57%, 32.14%, 53.57%, and 10.72%, respectively. The overall median progression-free survival (PFS) time was 6.43 months, and the median overall survival (OS) time was 11.43 months. The 1- and 2-year survival rates were 46.79% and 25.23%, respectively. Univariate analysis showed tumor response rate was an independent prognostic factor for PFS. The Cox proportional hazard model suggested the tumor response rate and prescribed dose were the independent prognostic factors for PFS. In addition, the independent prognostic factors for OS included tumor response rate, tumor diameter, and tumor volume. The common acute radiotherapy toxicities included gastrointestinal discomfort, radiation-induced liver damage, and myelosuppression. ConclusionIMRT is a safe and effective option for the LHCC patients who are unsuitable for surgery or in the cases that other local therapies fail.

Humans ; Lung ; Pneumoconiosis/diagnostic imaging* ; Radiography ; Tomography, X-Ray Computed

Objective To improve the fixation method of indwelling needle inserted in the tail vein in rats to keep the inserted needle for a longer indwelling time .Methods One hundred healthy Wistar rats ( age 5~6 months, male:female=1:1) were randomly divided into two groups: the experimental group and the control group .Rats in both groups received the same tail vein indwelling needle puncture and cannulation .The control group got the traditional fixation , namely fixed by sticking with 3M transparent dressing paste .The experimental group received in addition to the traditional fixation, a 0.1 mm-thick aluminum tube placed outside the needle fixation site .The indwelling time in the rats were recorded and analyzed .Results The indwelling time was (166.86 ±9.03) h and (20.24 ±5.04) h in the experimental and control groups, respectively (t =68.546, P<0.01).Conclusions Our improved method is safe and reliable.It can prolong the indwelling time of the punctured intravenous indwelling needle , and provides a useful rat model in studies on the complications of intravenous indwelling needle kept for a longer time .

Objective To investigate the effects of sodium butyrate(NaBT) on proliferation of human pancreatic cancer cell line ASPC-1 and explore the possible mechanism. Methods The methylthiazolyl tetrazolium assay (MTT) method was used to detect cell proliferation and draw a curve. The cell apoptosis and cell cycle were determined with flow cytometry. Western blot was used to study the effect NaBT on the pancreatic cancer cells and explore its mechansim. Real-time PCR was employed to assess the expression levels of p53, p21, bcl-2 and cell cycle regulation gene p21. Results After incubation with different concentrations of NaBT for 24 to 72 h, ASPC-1 cell proliferation was inhibited dramatically. NaBT induced an increase of G0/G1 phase cells and a significant decrease in the ratio of S phase cells. The expression of p21 and bax was up-regulated at protein and mRNA level. The expression of bcl-2 was down-regulated at protein and mRNA level. There was no significant difference in the expression of p53 at protein and mRNA level. Conclusion TSA-induced growth inhibition is associated with a block in the G0/G1 phase and apoptosis, which may occur through down-regulating the expression of apoptosis gene bcl-2 and up-regulating the expression of cell cycle regulation gene p21and pro-apoptotic gene bax.

OBJECTIVETo evaluate the expression of FBXW7 in esophageal squamous cell carcinoma (ESCC) and to explore the correlation of FBXW7 expression with clinicopathologic features and prognosis of ESCC.

METHODSNinety cases of ESCC and twenty cases of tumor-adjacent normal tissues and forty intraepithelial neoplasia tissues were detected by immunohistochemistry methods. The expression of FBXW7 in 40 surgical ESCC tissues and tumor-adjacent normal tissues was detected by Western blotting and RT-PCR. The relationship between the expression of FBXW7, clinicopathological characteristics and prognosis was analyzed.

RESULTSThe positive rate of FBXW7 protein was significantly lower in the ESCC and high-grade intraepithelial neoplasia tissues than in the tumor-adjacent normal tissues and low grade intraepithelial neoplasia tissues (40.0% and 33.3% vs. 85.0% and 77.3%, χ² = 21.923, P < 0.001). The FBXW7 protein was down-regulated in ESCC tissues (P < 0.05), whereas the FBXW7 mRNA was down-regulated in ESCC tissues compared with that in the paracancerous tissues. The expression of FBXW7 was significantly correlated with TNM stage, degree of differentiation, invasion depth and lymph node metastasis (χ² =9.643, 14.908, 16.294, 10.222, respectively, P = 0.002, 0.001, 0.000, 0.001). In the ninety patients, the 5-year survival rates of cases with positive and negative expression of FBXW7 protein was 67.6% and 39.3%, respectively (χ² =6.699, P = 0.01).

CONCLUSIONSThe results of this study demonstrate that FBXW7 expression is significantly declined in ESCC and high grade intraepithelial neoplasia tissues, and is closely correlated with poor prognosis of this disease. FBXW7 as a tumor suppressor gene may play an important role in the carcinogenesis, development and metastasis of ESCC. These results suggest that FBXW7 may become a valuable marker for the severity and prognosis in ESCC.

Biomarkers, Tumor ; metabolism ; Blotting, Western ; Carcinoma, Squamous Cell ; diagnosis ; metabolism ; Cell Cycle Proteins ; genetics ; Down-Regulation ; Esophageal Neoplasms ; diagnosis ; metabolism ; F-Box Proteins ; genetics ; F-Box-WD Repeat-Containing Protein 7 ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Neoplasm Staging ; Prognosis ; RNA, Messenger ; Survival Rate ; Ubiquitin-Protein Ligases ; genetics

Objective: To establish the method of dissolution determination for evaluating and improving the quality of Xindakang sugar coated Tablets. Xindakang Sugar coated Tablets from three enterprises were investigated. Methods: To adopt 900mL0.5% Tween80 H 2O as dissolvent and rotating basket method at 100r?min -1 , the cumulative dissolution percentage was determined by UV. The dissolution parameters was obtained by Weibull distribution model and dealed with one way ANOVA. Results: The significant differences in dissolution parameters(T d, T 50 ,m)( P

ObjectiveTo investigate the effects of insulin on the proliferation and invasion of human pancreatic cancer cells PANC1,and on its HIF-1α,VEGF expression.MethodsPANC1 was pretreated with insulin of different concentrations (0.1,1,10,100 nmol/L).The proliferation of PANC1 was tested by MTTmethod,and transwell assay was used to test the invasion ability of PANC1.HIF-1α,VEGF and PCNA protein expression was assessed by Western blots,and HIF-1α,VEGF mRNA was detected by real-time PCR.Results Insulin could increase the proliferation of PANC1 in a dose-dependent manner (p ＜0.05 ),and increase the expression of HIF-1α,VEGF protein.After 100 nmol/L insulin treatment for4 d,the PCNA protein expression in the insulin group was significantly higher than that in the control group (1.196 ±0.014 vs 1.157 ±0.013,P ＜ 0.05).The cancer cells passed through the chamber in insulin group were much more than that in the control group ( 141.0 ± 2.1 vs 89.0 ± 1.4,P ＜0.05 ).The expression of HIF-1α protein was significantly increased (1.139 ±0.020 vs 0.598 ±0.013,P ＜0.05),while there was no significant change of HIF-1αmRNA expression.Both the expression of VEGF protein and mRNA were significantly increased (1.011 ± 0.023 vs 0.627 ± 0.013 0.970 ± 0.016 vs 0.350 ± 0.01 3,P ＜ 0.05 ).Conclusions High insulin microenvironment could enhance the proliferation and invasion of PANC1 cells by up-regulating the expression of HIF-1α and VEGF.

ObjectiveTo evaluate gallbladder conserving gallstone removal and polyps resection using combination laparoscopy,hard gallbladder endoscopy and soft choledochoscopy.MethodsClinical data of 122 patients with cholecystolithiasis or polyps undergoing removal of calculus (polyps) and preservation of gallbladder were analyzed retrospectively.ResultsGallstones in 56 patients and polyps in 24 cases was removed or resected successfully by laparoscopy and hard gallbladder endoscopy; In the remaining 34 cases stones were completely removed by combination soft choledochoscopy; 8 cases were converted to laparoscopic cholecystectomy.Romoved stone was single in 25 cases and multiple in 65 cases,with the number ranging from 1to 52,the diameter of stone ranged from 0.2 cm to 3.2 cm.In the 24 gallbladder polyps,7 cases were single,17 cases were multiple,the diameter of polyp ranged from 0.8cm to 1.2 cm.The operation time was 40-125 (78) min. The mean hospitalization was 4 days. No intraoperative and postoperative complications occurred.All patients were followed up for 1year.Gallstones recurred in 3 cases,and the recurrence rate was 3.06％. ConclusionsLaparoscopy combined with hard gallbladder endoscopy and soft choledochoscopy for removing calculi (polyp) and conserving gallbladder is safe and feasible.

This study examined whether insulin-stimulated hypoxia-inducible factor 1alpha (HIF-1alpha) expression plays a crucial role in promoting the proliferative vitality and invasive capability in human pancreatic cancer cells. PANC-1 cells were divided into three groups: Control group, insulin group and insulin+YC-1 (a pharmacological inhibitor of HIF-1alpha) group in terms of different treatments. Cells in the insulin group or insulin+YC-1 group were treated with insulin (0.1, 1, 10 and 100 nmol/L) alone or combined with 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole (YC-1, 0.1, 1, 10 and 100 mumol/L). HIF-1alpha mRNA and protein expression in PANC-1 cells was determined by real-time RT-PCR and Western blotting respectively. Cell proliferation and invasion were measured by using growth curve and invasion assay, respectively. Western blot analysis demonstrated that insulin dose-dependently increased the HIF-1alpha protein expression, and YC-1 could dose-dependently block this effect. However, neither insulin nor YC-1 altered HIF-1alpha mRNA levels in PANC-1 cells. Moreover, insulin could enhance the proliferation and invasion of PANC-1 cells, while YC-1 could weaken this effect. It was concluded that the malignant proliferation and local invasion of pancreatic cancer cells may be related to high-insulin microenvironment. The tumor biological behavior change resulting from high-insulin microenvironment may be associated with the increased expression of HIF-1alpha protein.