Objective To explore the performance of serum AFP-L3 and PIVKA-Ⅱ in differential diagnosis of benign and malignant liver disease in high risk population.Methods The serum levels of AFP-L3 and PIVKA-Ⅱ in 48 patients with primary hepatic carcinoma,43 patients with cirrhosis and 81 patients with chronic hepatitis B were analyzed retrospectively.Results There was no statistically different significance among the median levels of serum AFP in primary hepatic carcinoma patients,cirrhosis patients and chronic hepatitis B patients (x2=4.014,P=0.134).Both median level of AFP-L3 and PIVKA-Ⅱ in primary hepatic carcinoma patients were higher than cirrhosis patients and chronic hepatitis B patients (x2 =33.93,52.33,both of P values were below 0.001).The specificity (92.74％) of AFP-L3 and the sensitivity (79.17％) of PIVKA-Ⅱ were all higher.The accuracy (84.88％) of combined detection in series was the highest,with its 47.92％ of sensitivity and 99.19％ of specificity.Conclusion Combined detection PIVKA-Ⅱ and AFP-L3 series will help to differential diagnosis of benign and malignant liver disease in high risk population.

Objective To investigate the mechanism of platelet inhibitor from Agkistrodon halys venom (AHV-PI) on platelet aggregation. Methods Protein kinase Akt phosphorylation levels in platelet were measured by Western blot. XS-1000I blood cell counter was used for platelet count. The plasma content of 5'-NT and platelet membrane GPIb were determined by Enzyme-Linked Immunosorbnent Assay (ELISA). The effect of AHV-PI on binding rate between the fluorescence labeled monoclonal antibody CD61 (FITC-CD61) and platelet membrae glycoprotein Ⅱb/Ⅲa (GPⅡb/Ⅲa) was observed by flow cytometry (FCM). Results AHV-PI can reduce the level of Akt-phosphorylation level and the number of platelet. AHV-PI can increase the content of 5'-NT in plasma, reduce the expression of platelet GPIb. Flow cytometry displayed AHV-PI can not affect the rate of combination between platelet GPⅡb/Ⅲa and FITC-CD61. Conclusion The mechanism of inhibition of platelet aggregation may be inhibit protein kinase Akt phosphorylation to block the signal transduction pathway of Akt. Limit cell grouth and reduce platelet number, also it may be related to its 5'-NT activity, it can degradate ADP to prevent the formation of platelet thrombus.

Objective To evaluate the diagnosis and treatment of cystic renal cell carcinoma and to improve the preoperative diagnosis and curative rate of the disease. Methods Ten cases of cystic renal cell carcinoma were retrospectively analyzed in the aspects of imaging and pathologic characteristics. There were 7 males and 3 females with average age of 56 years old (ranging from 38--74 years old) in this study. There were 3 cases complained of sore waist, 7 cases were found renal masses in annual physical examination and 2 cases had the history of renal cysts. The cyst diameter was 3.5 8.2 cm. Six cases had been diagnosed with ultrasound and 7 cases had been diagnosed with CT scan pre-operatively. Eight eases were diagnosed with frozen section during operation. All the 10 cases accepted radical nephreetomies. Results The post-operative histological diagnosis showed that there were 9 cases of clear cell carcinoma and 1 case of granular cell carcinoma. The pathological character istics were tumor necrosis of renal cell carcinoma in 6 cases, multilocular cystic renal cell carcinoma in 2 cases and carcinoma in renal cyst in 2 cases. Eight patients followed up from 6 months to 5 years. Six patients were still alive (mean 28.5 months). Conclusion The keys to improve the diagnosis and curative rate of the cystic renal cell carcinoma are paying attention to the pre-operative imaging study, the intra-operative frozen section examination and histopathology results.

Objective To establish whether Wnt-signaling pathway plays a role in mice β-cell function and/or survival in vitro. Methods Mice NIT-1 beta cells were cultured in media with glucose concentration of 33.3 mmol/L and the cytokines interleukin-1β, interferon-γand tumor necrosis factor-α with or without the addition of purified Wnt3a protein in vitro. Subsequently, β-cell apoptosis by Tunnel and flow cytometry, and β-cell proliferation by BrdU were analyzed. Total RNA was extracted to measure gene expressions by real-time PCR.Results Incubations of NIT-1 cells with high glucose and cytokines resulted in an increase in β-cell apoptosis and decrease in β-cell proliferation (P<0.01). In contrast, treatment with Wnt3a protein protected β-cell from glucose and cytokines-induced apoptosis through up-regulating the expressions of above Pitx2、 TCF7L2. Conclusions Wnt-signaling regulates the proliferation of pancreatic β-cell, and protectes β-cell from glucotoxicity and cytokine toxicity with respect to proliferation and apoptosis.

Objective To investigate the expression of peroxisome proliferator-activated rec eptor γ (PPARγ) and glucokinase (GK) induced by Wnt signaling pathway in mice NIT-1 β-cells,and to explore the interaction between PPARγ and Wnt signaling pathways.Methods Recombinant Wnt3a protein was applied to NIT-1 beta-cells to activate Wnt signaling pathway.The expression of PPARγ was determined by real-time PCR and Western blotting.The expression of GK was determined by real time PCR.Results Wnt3a rapidly activated Wnt/β-catenin/TCF signaling pathway,and increased PPARγ and GK mRNA expression by 41.2％ and 65.0％ in NIT-1cells,with PPARγ protein expression increasing by 97.8％ (P＜0.01).These effects were abrogated by Wnt and PIK3 inhibitors,dickkopf 1 and wortmannin treatment (P＜ 0.01).Conclusions PPARγ and GK can be upregulated by Wnt singnaling,and the effects might partially be PI3K-dependent.

OBJECTIVE To apply a low temperature hydrogen peroxide gas plasma sterilization for the heat intolerant orthopedic instruments.METHODS A comparison of the sterilizing effects of the orthopedic(instruments) was made among the low temperature hydrogen peroxide gas plasma sterilization,the ethylene oxide sterilization and the 2% glutaraldehyte(GTA).RESULTS Twelve hours were spent in the sterilization to the undurable(orthopedic) instruments when the ethylene oxide was used,and 10h were needed using the 2%GTA.but,55 min were enough using the low temperature hydrogen peroxide gas plasma.CONCLUSIONS The low temperature(hydrogen) peroxide gas plasma sterilization is applicable to rapidly sterilizing the heat undurable orthopedic(instruments) characteristically with the high effects,quick speed and convenient.

Objective To investigate the feasibility of recombinant adeno-associated virus encoding secrected forms of human acidic fibroblast growth factor transfecting endothelial progenitor cells.Methods sp-haFGF was obtained through combining signal peptide sequence of FGF-4 with native aFGF gene by PCR.sp-haFGF was cloned into AAV vector plasmid pAAV-IRES-hrGFP.Recombinant AAV encoding sp-haFGF was packaged through co-transfecting HEK293 cells with plasmid sp-haFGF-pAAV-IRES-hrGFP,pAAV-RC and pHelper.Ex vivo cultured EPCs were infected with concentrated rAAV.Expression of green fluorescent protein(GFP) in infected EPC was observed by fluorescence microscope and expression of sp-haFGF in EPCs was verified by RT-PCR and western blot analysis.Results Recombinant AAV encoding sp-haFGF was obtained.After EPCs being infected with rAAV,green fluorescence was found in about 20～30% EPCs,gene of(560 bp) was generated from EPCs by RT-PCR method,and(sp-haFGF) protein was detected in infected cells.Conclusion EPC was efficiently infected by rAAV encoding(sp-haFGF) and(haFGF) was expressed by EPCs,which establishes a basis for therapeutic angiogenesis by transgenic EPCs transplantation.

Objective To study the toxic effect of exotoxin A of Pseudomonas aeruginosa on keratocytes.Methods Three-dimensional gels of type I collagen containing rabbit keratocytes were incubated in the presence of different concentrations of exotoxin A(0.1,1.0,10 mg?L-1),cultivated for 24 h at 37℃,the change of keratocytes in morphology was observed under the light microscope,and the amount of lactate dehydrogenase(LDH) was measured by enzyme linked immunosorbent assay(ELISA).Results The LDH contents in different concentrations(0.1,1.0,10 mg?L-1)of exotoxin A groups were higher than that in the group without exotoxin A(P