Objective To study the influence of dihydromyricetin on the oxidation and non-enzymatic glycosylation in impaired glucose tolerance rats.Methods Animal model of impaired glucose tolerance rat was built by intragastrical(i.g.) injection of D-galactose generally.After 8 weeks intragastrical(i.g.) injection of dihydromyricetin(experimental group) and metformin (positive control),the levels of fasting blood glucose and tow-hour postprandial blood glucose,the insulin level and its resistance index,the levels of glycosylated hemoglobin (GHb) and advanced glycation end-products(AGEs) were detected,and the activities of superoxide dismutase(SOD) and glutathione peroxidase (GSH-Px),the content of malondiadehyde(MDA) were determined respectively.Results Compared with model group,dihydromyricetin decreased the level of two-hour postprandial blood glucose (P0.05).Conclusion Dihydromyricetin can significantly improve the state of impaired glucose tolerance rats; the mechanism may be related to its effect of improving the antioxidant activity of GSH-Px,inhibiting non-enzymatic glycation,lowering body oxidative stress and insulin resistance and promoting the use of blood glucose in peripheral organizations.

Objective To study the effects of the HBV x gene (HBx) on the biological characteristics and the expression of DNA repair enzyme hMTH1 mRNA of the L02/HBx transgene cell model. Methods Light microscopy was used to observe the morphologic characteristics of gene-transfected cell strain Lff2/HBx that stably expressed the HBx protein and the control groups of L02 and L02/PcDNA3.1. The changes of L02/HBx on the proliferation, cell cycle and apoptosis were observed by MTT assays and flow cytometry analysis respectively. Moreover, the malignant transformation of L02/HBxwas assayed by colony formation in soft agar and the expression of DNA repair enzyme hMTH1 mRNA was assayed in each group by real-time qPCR. Results Inversion phase contrast microscope showed that the morphologic characteristics of L02/HBx had changed obviously compared with control groups. The MTT showed that L02/HBx proliferated more quickly and flow cytometry analysis indicated that HBx could accelerate the progression of cell cycle and inhibit apoptosis. Colony formation in soft agar demonstrated that the rate of colony formation of L02/HBx was remarkably higher than the L02 and the L02/peDNA3. 1 cells (P<0. 05). The real-time qPCR detection showed that the expression of hMTH1 mRNA in L02/HBx was significantly higher than that in the control groups ( P < 0. 05 ). Conclusion HBx could play an important role in the malignant transformation of L02/HBx and the over expression of hMTH1 mRNA.

Transcatheter arterial chemoembolization (TACE) was the preferred method of non-operation treatment for hepatocellular carcinoma (HCC).Radical resection of HCC remains difficult,extrahepatic metastasis was not easy to deal with,and repeated treatment aggravated the liver injury,so the long-term efficacy was poor.Sorafenib could control tumor angiogenesis and block the proliferation of tumor cells.A male patient with primary HCC and in the stage Ⅱb according to the Chinese clinical liver cancer staging system was treated by TACE combined with sorafenib and antiviral treatment in Henan Cancer Hospital.After the treatment,the intrahepatic lesions were inactive,and the pulmonary metastasis was partially relieved.The patient was followed up till May 2012,and the survival time was 39 months.The hepatic function was normal,and the hepatitis B virus (HBV) replication was negative.No intervention treatmentrelated complications were detected,and the KPS score was 100.

A systematic method of isolating and culturing human bone mesenchymal stem cells (hMSCs), and inducing them to differentiate into neuron-like cells in vitro was established. The hMSCs were isolated from bone marrow with the lymphocyte-separating medium, cultured and expanded in vitro, and induced after addition of compound neuro-revulsants. The morphological changes of hMSCs were observed, and the expression of surface markers in induced hMSCs was immunocytochemically identified during induction period. The hMSCs could be separated, cultured and expanded in vitro. After induction by compound neuro-revulsants for 48 h, the changes of neuron-like cells, such as cellular shrinkage and neurite growth, were observed in some cells. The immunochemical staining revealed nestin (+) or NF (+), and GFAP (-). It was concluded that hMSCs were successfully cultured and induced to differentiate into neuron-like cells.
Bone Marrow Cells/*cytology ; Cell Culture Techniques ; Cell Differentiation/*physiology ; Cells, Cultured ; Mesenchymal Stem Cells/*cytology ; Neurons/*cytology

Objective To assess the therapeutic outcomes of transcatheter arterial chemoembolization combined with percutaneous injection of chemoembolization agent intra-portal vein tumor thrombosis for primary hepatic carcinoma accompanied by portal vein tumor thrombus. Methods Thirty patients with primary hepatic carcinoma accompanied by portal vein tumor thrombosis of type Ⅱ and type Ⅲ were randomly divided into two groups. The Child-Pugh ratings (class A and B) of group A and B were 9 vs 9 (class A) and 5 vs 7 (class B) respectively (χ~2 = 0.201, P > 0.05). The constitution of Type Ⅱ and type Ⅲ portal vein tumor thrombus in group A and B were 8 vs 9 and 6 vs 7 respectively (χ~2 =0.002, P>0.05). The median values of ALT, TBIL, ALB and AFP in group A and B were 58.7U/L vs 70.5 U/L (W=191.5, P>0.05), 21.4 μmol/L vs 21.7μmol/L (W=203, P>0.05), 35.3 g/L vs 37.5 g/L (W = 214, P > 0.05) and 680 μg/L vs 873 μg/L (W = 179. 00, P > 0.05) respectively. Group A was treated with transcatheter arterial chemoembolization (TACE) using emulsion made up of adriamycin, cisplatin, mitomycin and ultraliquidlipiodol plus percutaneous injection of chemoembolization agent intra-portal vein tumor thrombosis using emulsion consisted of cisplatin and ultraliquidlipiadol, while group B was treated with TACE only as a control group. Survival analyses were performed via the Kaplan-Meier test in SPSS11.5 with the log-rank tests with an threshold of 0.05. Results The 3, 6 and 12 months survival cases of group A and B were 11 vs 10, 10 vs 3, and 7 vs 0 respectively. The median survival time of group A and group B were 14.0 months and 4.0 months respectively. The difference of the two groups was significantly (χ~2 =11.728, P<0.01). There was no severe side-effect related to therapy in both groups. Conclusion Comparing with the control group, TACE combined with percutaneous injection of chemoembolization agent intra-portal vein tumor thrombosis could significantly prolong the median survival time of patient with primary hepatic carcinoma accompanied by type Ⅱ and type Ⅲ portal vein tumor thrombosis.

ObjectiveTo investigate the effect of living donor right liver graft transplantation (LDLT) with middle hepatic vein (MHV) on the early congestion and regeneration of the donor remnant liver.MethodsBetween August 2008 and August 2009,28 LDLT were performed with 11 LDLT without MHV (group A) and 17 LDLT with MHV (group B).The donor operative time,intraoperative blood loss,postoperative hospital stay,bilirubin,INR,and ALT level were recorded in detail.We measured the volume of remnant liver by means of CT scan 2 weeks after operation and compare the degree of congestion and regeneration of the remnant liver between the two groups.ResultsThere were 10 cases in group B and 0 cases in group A suffering from congestion at segment Ⅳ,and the difference was significant(P =0.006).In group B,6 cases in type Ⅰ and 4 cases in type Ⅱ developed congestion at segment Ⅳ,and the difference was significant(P=0.035).Two weeks post operation,the volume of segment Ⅳ in group B was smaller than in group A(P=0.005).The regeneration rate of segment Ⅳ in group B was smaller than in group A (P =0.007),on the contrary,the regeneration rate of segment Ⅰ - Ⅲ in group B was larger than in group A( P =0.008 ).But the regeneration rate of remnant liver was the same in both groups (P =0.63 ).ConclusionsThe right lobe hemihepatectomy with MHV does not damage the early liver function of the donor significantly.The segment Ⅳ of the remnant liver suffered from congestion and impeded the regeneration,but was compensated by the regeneration of segments Ⅰ - Ⅲ.

Objective To improve the orotracheal intubation verifying technique and reduce the complication of backflow in rat experiment.Methods A new position evaluation of anti-backflow device was designed and made of safety IV catheter and closed IV catheter system.60 adult male Sprague Dawley rats 216±20 g were randomly assigned to two groups: group A (n=40) for verifying placement, group B (n=20) for anti-backflow test.Group A was further divided into group A1 using self-designed positioning device, group A2 using aerosol, group A3 taking cotton fiber for positioning judgment.The group B was divided into two subgroups, B1 and B2, counting escaped bubbles as a means of positioning observation, the difference is that group B1 using frustum of a cone shape anti-backflow device, while the group B2 using common airway tube.Routine endotracheal intubation was performed to observe and record the time of positioning, the location of exhalation phase, and the length of inspiratory phase countercurrent water column.The group A1 further performed tracheotomy under direct vision clearly to confirm the anatomic positioning status.Results During the exhalation cycle,three or more bubbles were observed to escape continuously, indicating that the intubation tube was properly placed and open in the airway.Positioning time: It took 1.75±1.02 respiratory cycles in group A1,3.30±0.95 respiratory cycles in group A2 and 4.10±0.99 respiratory cycles in group A3 to complete the assessment the positioning status.There was no statistically significant difference between groups A2 and A3 (P> 0.05).The time needed for group A1 was significantly shorter than that of groups A2 and A3 (P < 0.01).The longest countercurrent water column length in group B1 was 3.23±0.53 cm, and 8.48±1.01 cm in the group B2.Conclusions The new designed anti-backflow positioning evaluation device is a simple and convenient appliance to evaluate the location of orotracheal intubation in rat experiment.It can effectively improve the positioning efficiency and has practical application value.

Objective To assess the therapeutic effect of combination chemotherapy of gemcitabine and cisplatin by double way plus implantation of radioactive seed 125I implantation in treating stage Ⅲ non-small cell lung cancer. Methods Sixty cases with stage Ⅲ non-small cell lung cancer were randomly divided into two groups with random number table. In group A (in interventional treatment group, n = 30),the gemcitabine 1000 mg/m2 and one third of the cisplatin 100 mg/m2 was given using seldinger technique for transcatheter bronchial arterial infusion chemotherapy on day 1. Two-thirds of the cisplatin 100 mg/m2 was infused in veins on day 2 and 3. The gemcitabine 1000 mg/m2 was infused in veins on day 8, 21 days for a period. In group B (interventional - 125I groups), the method of combination chemotherapy of gemcitabine and cisplatin was the same as in Group A. After ten days of arterial perfusion, 125I seeds were implantated, 21 days for a period. All patients received at least 2 cycles. The imaging evaluation of patients after treatment standards included complete remission (CR), partial remission (PR), stable (SD),progressive disease (PD), effective rate (CR + PR)/30 and clinical benefit rate (CR + PR + SD)/30.Non-parametric rank sum test was used to compare short-term effect of the two groups treatment of two cycles.x2 test was used to compare year survival, Kaplan-Meier method was used to calculate median survival,log-rank test method was used to difference between the groups. Results In group A, there were 17 PR,9SD and 4 PD. The overall response rate was 56. 7% (17/30) and clinical beneficial rate was 86. 7% (26/30). In Group B, there were 2 CR, 21 PR, 7 SD. The overall response rate was 76.7% (23/30) and clinical beneficial rate was 100% (30/30). There was significant difference between the two groups (P =0. 036). In group A, the 1 year survival rate was 46. 7% (14/30) and the 2 year survival rate was 36. 7%(11/30), median survival time (MST) was 10 months . In group B, the 1 year survival rate was 76. 7%(23/30) and the 2 year survival rate was 63. 3% (19/30) , median survival time (MST) was 27 months.There was a significant difference between two group in 1 year survival rate (P = 0. 017), 2 year survival rate (P = 0. 039) and median survival time (P = 0. 006). Conclusion The treatment effects of Ⅲ stage non-small cell lung cancer by gemcitabine and cisplatin combination chemotherapy with double way plus radioactive seed 125I implantation was better than gemcitabine and cisplatin combination chemotherapy with double way.

Objective To construct wtp53/junB fusion gene and its eukaryotic expression vector in order to provide the basis for further application of polygene union therapy in hepatocellular carcinoma. Methods Polymerase chain reaction (PCR), reverse transcription-PCR (RT-PCR) and gene recombination techniques were used to construct the eukaryotic vector of pEGFP-C1-wtp53/junB fusion gene, which carries the enhanced green fluorescent protein (EGFP). The transfection of pEGFP-C1-wtp53/junB in hepatoma HepG2 cells was detected by the location of green fluorescence. Results The DNA sequence of wtp53/junB fusion gene was successfully cloned into the pEGFP-C1 plasmid and the sequence was the same as what we expected. Green fluorescence located on cell nucleus proved that pEGFP-C1-wtp53/junB was transfected into HepG2 cell line successfully. Conclusion We successfully constructed the eukaryotic vector of pEGFP-C1-wtp53/junB fusion gene, which carries the EGFP, and transfects it into human hepatoma cell nucleus. It may lay the basis for studying the synergetic effect of wtp53 and junB in hepatocellular carcinoma.

ObjectiveTo evaluate the effects caused by different doses of N-nitrosodiethylamine (DENA) on establishment of a multistep carcinogenesis model of liver cirrhosis in SD rats.MethodsFifty male SD rats were randomly divided into control group (n =5 ) and experimental group (n =45 ).Experimental group consisted of 3subgroups(high,median and low dosegroup ),which were intragastrically induced DENA once a week for 10 weeks,by using 70,50,35 mg/kg DENA solution,respectively.MRI was performed for inspecting pathological changes of rat livers in the following weeks.The mortality rates of different groups were calculated.The differences of the incidence of dysplastic nodule (DN)found in three groups were analyzed with Pearson Chi-square test.ResultsThe multistep carcinogenesis development from regenerative nodule (RN) to DN to hepatocellular carcinoma (HCC) was showed in the SD rat model.Two rats in median dose group and 4 rats in high dose group died.The incidence of DNs found in 3 different groups was 20.7％ ( 6/29 ),30.6％ ( 19/62 ),14.3％ (9/63),respectively,which showed no significant difference among three groups(x2 =4.901,P ＞ 0.05).However,significant difference existed between median and high dose group( x2 =4.811,P ＜ 0.05 ).Combined with the MRI presentation,the intragastric dose of 50 mg/kg could be more appropriate among three different doses.ConclusionsThe median dose DENA could induce an ideal multistep carcinogenesis model of HCC which is suitable for investigating DN on MRI.