To investigate the expression and clinical significance of p27kip1 protein in primary liver cancer, the expression of p27kip1 protein and the relationship with clinicopathological factors were studied in primary liver cancer by using SABC immunohistochemical staining in specimens of 40 cases of primary liver cancer and 20 cases of liver cirrihosis. Our results showed that positive expression rate of p27kip1 protein in primary liver cancer was 37.5% (15/40), which was lower than that in benign lesion of liver 80.0% (16/20, P<0.01). The expression level of p27kip1 protein in primary liver cancer showed significant differences in tumor size, Edmonson histological grade, portal invasion, lymph node metastasis, TNM stage (P<0.05, for all), but not significantly correlated with patient's age and histological types. Log rank test showed that the p27kip1 expression was significantly related with prognosis of the patients (P<0.05), and the prognosis of the patients with p27kip1 positive expression was markedly better than that of those with p27kip1 negative expression. It is concluded that the expression of p27kip1 was significantly related clinicopathological factors of primary liver cancer. p27kip1 protein may be used as a novel tumor marker for primary liver cancer.
Carcinoma, Hepatocellular/*metabolism ; Carcinoma, Hepatocellular/pathology ; Intracellular Signaling Peptides and Proteins/*metabolism ; Liver Neoplasms/*metabolism ; Liver Neoplasms/pathology ; Prognosis ; Retrospective Studies ; Tumor Markers, Biological/*metabolism

Objective To study the relationship between HAI and HBsAg, HCV in HCC, pericarcinomatous tissues. Methods The patterns of HBsAg and HCV in 100 cases of hepatocellular carcinoma(HCC) and their surrounding liver tissues were studied on paraffin-embeded sections with immunohistochemistry technique, and pericarcinomatous tissues were scored in Knodell’s histological activity index(HAI). Results The score of HAI in the group of co-infection of HBV, HCV is the highest in the four groups(12.62?3.88). The score of HAI in the group which is not infected by HBV, HCV is the lowest in the four groups(6.67?2.58). HBV, HCV virus infection were positively correlated with HAI(rs=0.39,P=0.0001). HBsAg and HCV were detected both in HCC and pericarcinomatous tissues. The positive rate of HBsAg in Pericarcinomatous Tissues(79%) was higher than that of in HCC tissues(23%). HCV expressions in HCC(15%) and pericarcinomatous tissues(23%) had no differences. Conclusions As for the tissues of liver cancer with virus infection background, the HAI is obviously higher than that without virus infection background. HBV, HCV virus infection were correlated with HAI significantly, perennial virusemia will aggravate pathological changes of liver tissue.

Objective To investigate the experimental condition of separate purification and directed differentiation potency of pancreatic stem cells of adult rats in vitro. Methods The pancreas of adult rats were digested by collagenase V in situ perfusion. Filtration was performed by 100 mesh filter. Ficoll 400 density gradient centrifugation was used to separate pancreatic stem cells and stem cells were for cultivation. Epidermal growth factor (EGF), fibroblast growth factor (bFGF) was added to medium for in vitro cultivation.Immunofluorescence staining method was used to detect the expression of CK19, Pdx-1, nestin, insulin and glucagon, and the positive cell rate was calculated. The differentiated cell was evaluated by dithizone (DTZ)staining; insulin excretion function was determined by optical enzyme labeled ELISA staining. Results The pancreatic stem cell obtained from the study could be cultivated in vitro for 8 generations. The expression of CK19, Pdx-1 and nestin of the tem cells were all positive, and the rate of positive cells were (88.6 ± 6.2)% ,(84.6 ±8.6)% and (81.3 ±7.5)%. The differentiated cell was in brownish red color by DTZ dye. After high concentration sugar stimulation, the expression of insulin secretion was increased in the supernatant.Conclusions This method can harvest highly purified, and large amount of pancreatic duct stem cell.Artificial induction may result directed differentiate for islet-like clusters and have insulin secrete function.

BACKGROUND: Glucose is an important factor on differentiation of pancreatic duct stem cells, it relates to the quantity and secretion function of insulin-producing cells after differentiation.OBJECTIVE: To compare the insulin secretion capacity of the differentiated rat pancreatic stem cells induced by various glucose concentrations.METHODS: Rat stem cells were isolated and purified from pancreatic duct cells using collagenase V and Ficoll-400. These stem cells were randomly divided into 10 groups. Every group was induced to culture, proliferate, differentiate and form insulin- producing cells in vitro. The differentiation of all groups was performed in medium with different concentrations of glucose. The immunofluorescence staining was used to identify the pancreatic duct stem cells. The electrochemical luminescence method was used to detect the insulin release from stem cell differentiated islets.RESULTS AND CONCLUSION: The stimulation index of glucose 20.6, 25.6, 30.6 mmol/L groups was higher than that in other groups (P < 0.05), but there was no difference between each two groups among these three groups (P > 0.05). The insulin releasing of glucose 15.6, 20.6, 25.6 groups was higher than that in other groups (P < 0.05), but there was no difference between each two groups among these three groups (P > 0.05). The best insulin secretion capacity of insulin-producing cells can be gained by controlling concentration of glucose as 20.6-25.6 mmol/L when pancreatic duct stem cells differentiated into insulin-producing cells.

BACKGROUND: Adipose-derived mesenchymal stem cells (ADMSCs) can improve the liver function of rats with liver failure, which illustrates the important research value in the field of tissue engineering and cell transplantation.OBJECTIVE: To evaluate the therapeutic potential of human ADMSCs in heart failure rats and to discuss the possible biological mechanisms involved.METHODS: Heart failure rats were randomized into model and ADMSCs groups, which were given normal saline or DAPI-labeled human ADMSCs (3.0×106) via the tail vein. At 1, 3, 7 days after transplantation, we detected the biochemical indexes for liver function in rats. At 3 days after transplantation, the serum levels of cytokines, such as tumor necrosis factor α and interleukin-10, were detected, the histomorphological changes in the liver were observed by hematoxylin-eosin staining, and the protein expression of proliferating cell nuclear antigen was detected by western blot. RESULTS AND CONCLUSION: We found that human ADMSCs could migrate to the liver and lung tissues in rats after the transplantation via the tail vein. At 1 and 3 days after transplantation, the levels of serum alanine aminotransferase and aspartate aminotransferase were significantly reduced in the ADMSCs group as compared with the model group (P< 0.05); furthermore, the secretion of tumor necrosis factor α and interleukin-10 was significantly suppressed at 3 days after cell transplantation (P < 0.05). The results of hematoxylin-eosin staining indicated a significant improvement in liver degeneration and necrosis. The expression of proliferating cell nuclear antigen protein in the ADMSCs group was significantly up-regulated compared with the model group. To conclude, human ADMSCs can inhibit the inflammatory reaction and up-regulate the expression of proliferating cell nuclear antigen, to promote the regeneration of liver cells and he recovery of liver function.

Objective To evaluate the effect of local arterial infusion (LAI) of medicine on the treatment of servere acute pancreatitis(SAP). Methods The clinical data of 85 cases of SAP were retrospectively analyzed, and divided into three groups according to the admitted time:Group A. 42 cases admitted from February 1982 to December 1993,treated mainly by operation. Group B. 23 cases, admitted January 1994 to Auguest 1996, treated mainly by non operation. Group C. 20 cases, from September 1996 to Auguest 2000. treated mainly by LAI. Results The secondary infection rate in group A, B and C were 47%(20/42),26%(6/23) and 10%(2/20) respectively. The mortality in group A ,B and C were 36%(15/42),22%(5/23) and 5%(1/20),respectively. The difference in the secondary infection rate and mortality between group A , B and group C showed obvious significance (P

BACKGROUND:Recent studies have shown that adipose-derivedmesenchymal stem cells (ADMSCs) not only have multilineage differentiation potential, but also exert an important role in blood sugar balance and hormone production.OBJECTIVE:To observe the differentiation potential of human ADMSCs (hADMSCs) into functional islet-like cells and the therapeutic effect of hADMSCs transplantation in diabetic rats.METHODS:PDX-1 gene was transfected into hADMSCs by adenovirus. Cell differentiation and insulin secretion were identified and detected by dithizone staining and ELISA, respectively. Twenty male Sprague-Dawley rats were randomly divided into control group (n=4), diabetes group (n=8) and transplantation group (n=8). Rats in the latter two groups were subjected to making diabetic models by 65 mg/kg streptozotocin injection. Afterwards, rats in the transplantation group were given PDX-1 transfected ADMSCs via the tail vein.RESULTS AND CONCLUSION:At 15 days after transfection, the number of insulin positive cells and insulin secretion were both increased significantly (P < 0.05). Fasting glucose levels in the transplantation group decreased significantly (P < 0.05), while the body weight increased significantly (P < 0.05). In the diabetic group, the fasting glucose level still maintained at a high level, and the body weight of rats was significantly decreased. These results implicated that PDX-1 gene could induce hADMSCs differentiating into functional islet-like cells. PDX-1 transfected ADMSCs transplantation is effective in treating diabetic rats, but the mechanism needs further study.

Objective:To investigate the impact of Notch signaling pathway on the migration of human hepatic carcinoma cells and the expression of E-cadherin and COX-2 in these cells.Methods:Cultured hepatic carcinoma cell lines (SMMC-7721,MHCC97H),and normal non tumor liver cell line (HL-7702) in vitro.Transwell cell was used to measure the cell's capacity of invasion and migration.Western blot was used to measure the expression level ofNotch1,E-cadherin,COX-2 protein.DAPT was used to block the Notch signaling pathway,and compared the ability of invasion and migration between hepatic carcinoma cell lines and normal non tumor liver cell line,and the change of expression level of E-cadherin and COX-2 protein in hepatic carcinoma cells.Results:The migration ability of SMMC-7721 cells and MHCC97H cells were higher than HL-7702 cells,the difference was statistically significant (P＜0.05);Compared to HL-7702 cells,the expression level of Notch1 and COX-2 in MHCC97H cells and SMMC-7721 cells significantly increased,the expression level of E-cadherin decreased significantly (P＜0.05);After DAPT treatment,the migration ability of SMMC-7721 cells,MHCC97H cells were weaker than the control group,the difference was statistically significant (P＜0.05);After DAPT treatment,the expression of COX-2 and Notch1 in SMMC-7721 and MHCC97H cells decreased significantly,while the expression of E-cadherin significantly increased (P＜0.05).Conclusion:Notch signaling pathway plays an important role in the process of liver cancer cell migration and invasion,and its mechanism is related to the expression of E-cadherin and COX-2.

Objective To evaluate the safety and efficacy of magnetic resonance-guided focused ultrasound surgery (MRgFUS) in treatment of symptomatic uterine leiomyoma among Chinese reproductive age women.Methods From April 2010 to April 2012,80 premenopausal women with symptomatic leiomyoma volunteered to participate in this prospective study in Department of Outpatient of Peking Union Medical College Hospital.Among 23 reproductive aged patients with size of uterus less than 16th gestational weeks,2.5 to 10 cm of diameter of myoma,less than 10 myomas and expressing symptoms clearly were treated by MRgFUS.Treatment data,non-perfused volume ratio (NPVR) and adverse events were recorded.After treatment,patients were followed up at 1 week,1,3,6,12 and 24 months,respectively.Patients at initial screening and each time of the follow-up filled out uterine fibroid symptoms quality of life (UFS-QOL),which include symptoms severity score (SSS) and health-related quality of life (HRQL).The volumes of leiomyoma and uterine were evaluated on MRI before and after the treatment (at 6 and 12 months,respectively).Before operation,routine blood test were performed on all patients,anemia patients at 3 months and 1 year after treatment were checked with blood test.Results (1) Treatment data and adverse events:the mean therapeutic temperature was (69 ± 7) ℃,the mean treatment time was (144 ±62) min,the mean NPVR was (62 ±23)％.Adverse events included mild erythema(1/23),abdominal cramp(8/23),vaginal discharge (5/23),and leg numbness (4/23).(2) The rate of secondary surgery:one patient was treated by myoectomy and hysterectomy within one year following up and 4 patients chose surgical treatment during the second-year follow-up.(3) Volume change:the volumes of leiomyoma before the treatment and 6,12 months after the treatment are 75.6 (P25 =43.8,P75 =128.9),52.3 (P25 =23.8,P75 =111.2),45.9 (P25 =26.3,P75 =71.7) cm3,respectively; and the volumes of uterine before the treatment and 6,12 months after the treatment are 270.0 (P25 =208.4,P75 =390.3),216.4 (P25 =151.1,P75 =290.0),200.0 (P25 =149.1,P75 =267.6) cm3,respectively.Both leiomyoma and uterine volumes decreased significantly after treatments (P ＜ 0.01).(4) UFS-QOL change:the symptoms severity score (SSS) before the treatment and 3,12 months after the treatment are (34 13),(22 ± 11),(19 ± 12),which decreased significantly (P ＜ 0.01).The health-related quality of life (HRQL) before the treatment and 3,12 months after the treatment are (74 ± 15),(82 ± 13),(89 ± 10),which increased dramatically (P ＜0.01).(5) Hemoglobin(HGB) change:eleven patients suffered from anemia before treatments,the mean HGB before treatment was (87 ±6) g/L and were (106 ± 14) g/L 3 months after treatment,(112 ± 10) g/L 12 months after treatment.The HGB was increased significantly after treatments (P＜0.01).Conclusions MRgFUS is a safe and effective non-invasive management for symptomatic uterine leiomyoma in short-term follow up.But there is additional treatment ratio after MRgFUS.

Objective To investigate the potential of pancreatic stem cells (PSCs) directed differentiation in vitro, and to evaluate the effects of differentiated PSCs allograft on the treatment of diabetes.Methods The PSCs of adult Wistar rats were separated and purified in vitro. The surface of PSCs was determined by immunofluorescence staining, and then it was stimulated by hepatocyte growth factor (HGF) and nicotinamide to induce directed differentiation. Dithizone dyeing was used to determine the islet-like cells after induction, and ELISA staining method was used to detect the insulin levels. Streptozotocin peritoneal injection was used to induce the diabetic rat mode. 40 rats were randomly allocated into pancreatic islet cells allograft group (experiment group) and placebo group. The serum insulin and glucose levels 1 d before transplantation and 1, 2, 3, 4 week after transplantation were measured. Results PSCs of adult Wistar rats were successfully obtained, and the expression of CK19, Pdx-1 and Nestin on cell surface was positive. Dithizone dyeing for directed differentiation cells showed brownish red color. The cells could express and secrete insulin after hyperglycaemia stimulation. The serum insulin and glucose levels 4 week after transplantation were (11.41 ±1.52) mU/L and (8.22 ± 2.7) mmol/L, which were (9.30 ± 1.56) mU/L and (12.23 ± 3.8) mmol/L in the placebo group, and difference was statistically significant (P＜0.05). Conclusions PSCs can be induced and directed differentiated in vitro into islet-like clusters with insulin secretion function. And its allograft has the potential for the treatment of diabetes.