Objective To establish a pancreatic cancer model in Sprague Dawley(SD) rats,and to study the distribution and the counts of myofibroblast(MF) in pancreatic cancer and non-cancerous pancreatic tissues.MethodsDirectly implanted DMBA into pancreas parenchyma of SD rats and established TSA intervening group and control group. The carcinogenesis of rats executed within 3～5 months were inspected by HE stain and microscopy for pathomorphological changes.Myofibroblast(MF) was stained by Heidenains method. Results (1)The prevalence of pancreatic cancer in experimental group was 48.7%(18/37), including 17 pancreatic ductal adenocarcinoma and 1 fibrosarcoma. The prevalence of pancreatic cancer in intervering group was 33.3%(12/36), including 11 pancreatic ductal adenocarcinoma and 1 fibrosarcoma. The maximal diameters of tumor mass of experimental group was higher than that of intervering group (P

ObjectiveTo study the expression of ABCG2, SFRP2, BRMS1 and HPA and detect their clinicopathologicalsignificancesinthebenignandmalignatntlesionsofthegallbladder.MethodsEnVisiom immunohistochemical method for determining the expressions of ABCG2, SFRP2,BRMS1 and HPA was used in paraffin-embedded sections of surgical resected specimens from gallbladder adenocarcinoma (n =108), peritumoral tissues (n =46 ), adenomatous polyp (n =15 ), and chronic cholecystitis ( n =35 ).ResultsThe positive rates of ABCG2 and HPA expression were significantly higher in gallbladder adenocarcinoma than that in peritumoral tissues, adenomatous polyp and chronic cholecystitis (P ＜ 0. 05 or P ＜ 0. 01 ) ; The positive rates of SFRP2 and BRMS1 expression were significantly lower in gallbladder adenocarcinoma than that in peritumoral tissues, adenomatous polyp and chronic cholecystitis(P ＜0. 05 or P ＜0. 01 ). The positive cases of ABCG2 and/or HPA as well as negative ones of SFRP2 and/or BRMS1in the benign lesions showed moderately-or severely-atypical hyperplasia of gallbladder epithelium. The frequency of samples with positive staining for ABCG2 and/or HPA in cases with small tumor volume (diameter ＜ 2 cm), no lymph node metastasis, and no invasion into surrounding tissues was significantly lower than that in cases with larger tumor volume (diameter＞ 2 cm ), lymph node metastasis, and invasion into surrounding tissues ( P ＜ 0.05 or P ＜ 0. 01 ). However, compared with ABCG2 and/or HPA, the expression of SFRP2 and/or BRMS1 showed an opposite correlation in these cases ( P ＜ 0. 05 or P ＜0. 01 ). Unitivariate Kaplan-Meier analysis showed that increased expressions of ABCG2 (P =0. 019) and HPA ( P =0. 016) or decreased expression of SFRP2 ( P =0. 019) and BRMS1 ( P =0. 008 )were associated with poorer overall survival, respectively. Multivariate Cox regression analysis showed that increased expression of ABCG2 (P =0. 018 ) and HPA ( P =0. 019) and/or decreased expression of SFRP2 (P =0. 015 ) and BRMS1 ( P =0. 011 ) were independently poor-prognostic predictors in gallbladder adenocarcinoma.ConclusionsThe expression of ABCG2, SFRP2, BRMS1 or HPA might be closely related to the carcinogenesis, clinical biological behaviors, and prognosis of gallbladder adenocarcinoma.

Objective To compare the size of ablation lesions created by normal saline enhanced radiofrequency ablation (NS-RFA) and dilute hydrochloric acid enhanced radiofrequency ablation (HCl-RFA), explore their affecting factors, and observe the morphological manifestations of the ablated lesions.Methods NS-RFA and HCl-RFA were performed on 30 excised porcine livers with 9 different combinations of durations (5, 10, 15, and 20 minutes), temperatures (83, 93, 103, and 113 ℃ ) and powers (20, 30,and 40W). For each ablated lesion, the longitudinal and transverse diameters were measured, and volumes calculated. Multifactor analysis of variance was used to analyze the affecting factors of the size of ablated lesions. Macroscopic and microscopic morphological characteristics of lesions were observed. Results ( 1 )NS-RFA lesion volumes under 9 combinations were ( 3.53 ± 0. 34 ), (6. 41 ± 0. 42 ), ( 10. 69 ± 0. 37 ),(11.40±0.51), (3.20±0.23), (6.59 ±0.50), (12.11 ±0.70), (11.12 ±0.52), (11.81 ±0. 64) cm3, respectively. HCl-RFA lesion volumes under 9 combinations were ( 11.97 ± 1. 00), (28.72 ±0.99), (59.45 ±1.33), (105.65 ±2.40), (13.64±0.60), (29.70±0.58), (59.22±1.32),( 57. 22 ± 3.99 ), ( 59. 74 ± 2. 18 )cm3, respectively. The size differences of ablation zones caused by different types of ablation ( F = 948.9 ) ( main factor), durations ( F = 269. 3 ) and temperatures ( F =214. 6) (covariates) were statistically significant (P ＜ 0. 01 ), whereas which caused by power ( F = 0. 2 )(covariate) was not statistically significant (P ＞ 0. 05 ). (2)At gross examination, all ablation lesions were elliptical in cross section and there were three zones in NS-RFA induced lesions and five zones in HCl-RFA induced lesions. At microscopic examination of NS-RFA induced lesions, a small amount of liver cell debris were found at the edge of zone Ⅰ , a few of deformed and ruptured liver cells in zone Ⅱ. The shape of the most of the liver cells in zone Ⅲ was normal. At microscopic examination of HCl-RFA induced lesions, a small amount of liver cell debris were found at the edge of zone Ⅰ , classical coagulation necrosis in zone Ⅱ and Ⅲ, widened hepatic sinusoids lossened junction of hepatocytes and some hepatocytes detached into sinusoids in zones Ⅳ. The liver cells in zone V were normalexcept a small amount of hepatoeytes with pyknosis, karyorrhexis and karyolysis. Condusion Compared with NS-RFA, HCl-RFA can produce lager ablation zones. The duration and temperature were the factors that affected the size of ablation zone. HCl-RFA lesions typically showed coagulation necrosis at microscopical examination.

Objective To construct a stably-performing and high-efficiency regeneration system of Gynostemma pentaphyllum and lay a foundation for the gene transformation of it.Methods The blades of five-leaf G.pentaphyllum were used as the explants and cultured in MS media with different portions of hormone to induce fascicled-bud,root and plantlet regeneration.Results The medium suitable for inducing the blade differentiation of G.pentaphyllum was MS+6-BA 1.0 mg/L and for the frequency of blade differentiation could reach 40%.The cultural medium MS+6-BA 1.0 mg/L was suitable for the sub-multiplication of fascicled-bud and the medium 1/2 MS for root inducement and the plantlet regeneration.Conclusion A stably-performing acceptor system of the direct differentiation for Agrobacterium tumefaciens mediated gene transformation of G.pentaphyllum blades is constructed.

BACKGROUND: Embryonic stem cells fESCs) develop from early blastular inner cell mass.Under proper condition,ESCs can maintain undifferentiated state in vitro,normal diploid nuclear type,and proliferative potential.In addition,ESCs possess multi-directional differentiation capacity and can differentiate into all sorts of cells of three germ layers. OBJECTIVE: To investigate the feasibility of directed differentiation of ESCs towards thyroid cells in vitro as well as related molecular expression. DESIGN,TIME AND SETTING: A cell-based observational experiment was performed at the Bank of Cord Blood,Second Hospital Affiliated to Sun Yat-sen University between January 2004 and December 2006.MATERIALS: Balb/c pregnant mice at pregnancy 12.5-14.5 days were used for preparation of embryonic fibroblast feeder layer.E14 mouse embryonic stem cell (ESC) strains were gifted by professor Xu from Harvard University.METHODS: Murine El4 ESCs were cultured in methylcellulose semisolid medium to form embryoid bodies (Ebs).These Ebs were transferred for further inductive culture with the stepwise addition of growth factors- thyrotropin (TSH),insulin and kalium iodidum (KI).The cultured thyroid cells of adult Kunming mice were taken as positive control.MAIN OUTCOME MEASURES: ①Cellular morphological change in the process of differentiation.② Detection of expression levels of TSHR,PAX8,TTF-2,and TIF-1 by immunofluorescence assay.③ Detection of expression levels of TSHR,PAX8,NIS,TPO,and Tg by reverse transcription-polymerase chain reaction (RT-PCR).RESULTS: The differentiated cells had clear boundary,exhibiting round,oval,shuttle-shaped,or polygon adhesive growth.On day 6 of inductive ifferentiation,the differentiated cells showed the expression of PAX8,NIS,TPO,Tg,and TSHR,the specific gene of thyroid ceils.On day 8 of inductive differentiation,the expression of TSHR,TIF-1,PAX8,and TIF-2 was detectable in the differentiated cells with morphous similar to thyroid cells.CONCLUSION: ESCs can differentiate towards thyroid cells under given inductive conditions.