Objective To study the inhibitory effect of intrasplenic injection of retroviral packaging cells encoding human interleukin-2 (hIL-2) and mouse interleukin-12 (mIL-12) fusion gene on the growth of chemically induced hepatocellular carcinoma in rats. Methods The retroviral vector GCIL12EIL2PN encoding hIL-2 and mIL-12 fusion gene was constructed. The retroviral packaging cell line PA317 transfected with the vector was injected into the spleens of rats with established chemically induced hepatoma on on the 90th day (early-stage treatment) or the 105th day (late-stage treatment). The survival time and toxic effect were observed. The serum mIL-12 and hIL-2 levels were assayed with ELISA, and the cytotoxicity of the natural killer (NK) cells was measured by means of a 51Cr-release assay using YAC-1 tumor cells as the target. Results The average survival time (after chemical induction) in the early-stage treatment rats and the late-stage treatment rats were 188.1?14.2 days and 168.5?13.6 days, respectively, in IL-12+IL-2 combination gene treatment group, and it was longer than that of IL-12 gene treatment group (168.2?13.4 days and 149.1?13.8 days, respectively, P

Objective To evaluate the feasibility,safety,and efficacy of laparoscopic radiofrequency ablation(RFA) for the treatment of hepatic cavernous hemangioma(HCH) in patients with liver cirrhosis.Methods A total of 15 patients with HCH and liver cirrhosis received laparoscopic RFA under general anesthesia between March 2001 and August 2005.There were 6 men and 9 women,with a mean age of 46.2?7.0 years.All the patients had complained of obvious symptoms of abdominal discomfort,pain,or fullness. The etiologic factor of liver cirrhosis was hepatitis B in 13 patients and hepatitis C in 2 patients.The Child classification revealed grade A in 10 patients and grade B in 5 patients.A total of 20 liver lesions located on the surface of the liver or adjacent to the gallbladder,with a mean diameter of 7.2?1.4 cm,were identified preoperatively in the 15 patients by ultrasonography,helical CT scans,or MRI.The platelet count was(31.2?10.4)?10~9/L.Co-morbidities included chronic calculous cholecystitis in 3 patients and diabetes mellitus in 2 patients.Laparoscopic ultrasonography and liver biopsy were routinely performed during the operation.Results Laparoscopic RFA was successfully performed in all the 15 patients and laparoscopic cholecystectomy was conducted simultaneously in 3 patients.The ablation time per lesion was 68.8?34.0 min,and the total operative time was 120.0?28.0 min.No severe complications,such as intraperitoneal hemorrhage,gastrointestinal injury,diaphragmatic injury,bile leakage,and liver function failure,developed during and after the operation.Complete tumor necrosis was achieved in all the 20 lesions(100%,20/20) on contrast-enhanced helical CT scans 1 month after the treatment.During a follow-up period of 5~31 months,symptoms completely disappeared in 13 patients and significantly subsided in 2 patients.Conclusions Laparoscopic radiofrequency ablation is a feasible,safe,and effective treatment for hepatic cavernous hemangioma complicating liver cirrhosis.

Objective To investigate the feasibility and efficacy of surgical resection combined with radiofrequency ablation (RFA) for multifocal hepatocellular carcinomas (HCC) in patients with cirrhosis. Methods A total of 18 patients with multifocal HCCs and liver cirrhosis was treated between August 2003 and January 2006. Forty-six hepatic lesions were identified preoperatively by ultrasonography, helical CT, or MRI. Ten patients were found as having 2 lesions, 6 patients having 3 lesions, and 2 patients, 4 lesions. Under general anesthesia, segmental hepatectomy for major lesions (with a 2cm resection margin) and RFA therapy for minor lesions were performed. Results The combination therapy was performed successfully in all the 18 patients. A cholecystectomy was performed simultaneously for gallstones in 2 patients, and a splenectomy with para-esophagogastric devascularization was performed for portal hypertension in 1 patient. The surgical resection time was 37.4?8.8 min, the RFA time per lesion was 25.6 ? 8.9 min, the total RFA time was 39.8 ?14.7 min, the total operative time was 152.4?30.8 min, and the intraoperative blood loss was 465.6 ? 171.0 ml. No severe complications, such as intraabdominal hemorrhage, gastrointestinal tract injury, diaphragmatic injury, and liver function failure, developed after operations. No residual tumor was found on the margin of surgical resection and a complete lesion necrosis was achieved in the RFA regions on contrast-enhanced helical CT scanning 1 month after the procedure. During a follow-up period for 6~31 months (mean, 15.5 months), 5 patients were diagnosed as having new malignant nodules and were given a percutaneous RFA therapy. Out of the 5 patients, one died from tumor recurrence and lung metastases, and two patients died from liver failure at 7 and 16 months after treatment, respectively. Conclusions Surgical resection combined with RFA therapy is a feasible, safe, and effective treatment for proper patients with multifocal HCCs and liver cirrhosis, preserving impaired liver functions to the greatest advantages.

AIM: To demonstrate the effects of telmisartan on the proliferation and apoptosis of U937 cells.METHODS: The proliferation ability of the U937 cells was assessed by CCK-8 assay and colony formation test with methylcellulose.The CD11b expression rate of the U937 cells was identified by flow cytometry.The apoptotic rate was analyzed by flow cytometry with Annexin V-PI double staining and Hoechst 33342 staining.The protein levels of cleaved PARP and cleaved caspase-3 were determined by Western blot.RESULTS: The results of CCK-8 assay confirmed that the viability of U937 cells was inhibited by telmisartan.The colony formation capacity of U937 cells was also significantly inhibited by telmisartan.The differentiation of U937 cells was induced by telmisartan with the expression of CD11b.The results of flow cytometry analysis with Annexin V-PI double staining and Hoechst 33342 staining identified that the apoptosis of U937 cells was induced by telmisartan in dose-dependent and time-dependent manners with the up-regulation of cleaved PARP and cleaved caspase-3 proteins.CONCLUSION: Telmisartan inhibits the proliferation and induces the differentiation of U937 cells.Telmisartan also induces the apoptosis of U937 cells through the caspase pathway.

AIM: To investigate the effect of c-Myc inhibitor 10058-F4 on human chronic myeloid leukemia ( CML) K562 cells and imatinib-resistant K562/G cells.METHODS: The protein expression of c-Myc was detected by Western blotting .Cell proliferation was evaluated by MTT assay and colony formation assay .PI staining was used to deter-mine the cell cycle distribution .Annexin V-PI staining was applied for apoptosis detection .RESULTS:Imatinib-resistant K562/G cells displayed lower sensitivity to imatinib than K 562 cells with high expression of c-Myc.Treatment with specific c-Myc inhibitor 10058-F4 inhibited the cell proliferation in a dose-and time-dependent manner , and K562/G displayed more sensitivity to 10058-F4 than K562 cells.10058-F4 also induced cell cycle arrest in G 0/G1 phase and induced apoptot-ic cell death in the 2 cells.Importantly, 10058-F4 suppressed the colony formation ability in K 562 and K562/G cells. CONCLUSION:c-Myc is a novel target to overcome imatinib-induced drug resistance , and c-Myc inhibitor provides a new approach in CML therapy .

Objective To assess the alterations of the systolic peak radial strain (RS) and the time to systolic peak radial strain(TRS) in ischemic myocardial segments with different extent of coronary artery stenosis using three-dimensional ultrasound speckle tracking imaging(3D-STI).Methods RS and TRS of 16 left ventricular segments were analyzed by 3D-STI in 87 patients,every left ventricular segment of all patients were divided into 5 groups according to coronary stenosis based on the results of selected coronary angiography:normal,≤25 ％,＞25 ％ - ≤50 ％,＞50 ％ - ≤ 75 ％,＞ 75 ％.All times were corrected by heart rate.Results In the coronary normal group,coronary stenosis extent ≤25％ group and coronary stenosis extent ＞25％ - ≤50％ group,the variance of RS was non-significant( P ＞0.05).Compared with the coronary normal group,coronary stenosis extent ≤25％ group and coronary stenosis extent ＞ 25％ -≤ 50％ group,RS was decreased in groups of coronary stenosis ＞50％ - ≤75％ and ＞75％.Between the coronary normal group and coronary stenosis extent ≤25 ％ group,the variance of TRS was non-significant( P ＞0.05).Compared with coronary normal and coronary stenosis extent ≤25 ％ group,TRS was increased in groups of coronary stenosis ＞25％ - ≤50％,＞50％ - ≤75％,＞75％.Compared with group of coronary stenosis ＞ 50％ -≤ 75％,TRS was shorter in group of coronary stenosis ＞ 75％,some variance was significant( P ＜0.05).Conclusions The RS was decreased along with the coronary stenosis increase,but TRS was increased along with the coronary stenosis increase.There is a tendency that TRS was decreased when the coronary stenosis is more than 75 ％ compared with the coronary stenosis ＞50％ - ≤75 ％ group.3D-STI can access the regional radial systolic function of the ischemic myocardial segment.

AIM: To detect the protein expression of TIMP3 and RUNX3 in bone marrow mononuclear cells (BMMCs) from acute leukemia (AL) patients and to investigate the relationship between the methylation status of genes and their expressional levels. METHODS: Protein expression of TIMP3 and RUNX3 in 50 samples of BMMCs and 10 samples of peripheral blood mononuclear cells (PBMCs) from healthy volunteers was detected by Western blotting. The prognostic factors related to AL and data from methylation specific polymerase chain reaction were also analyzed. RESULTS: The expression level of RUNX3 with methylation was less than that without methylation in BMMCs from AL patients. The complete remission (CR) rate was related to RUNX3 expression and blasts in bone marrow (BM). BMMCs from patients with silencing of RUNX3 and higher blasts in BM had a lower CR rate. CONCLUSION: Absence of RUNX3 protein expression resulting from methylation of RUNX3 promoter probably plays a role in the pathogenesis of AL and is of value in prognosis. No relationship between methylation of TIMP3 promoter and the pathogenesis of AL is observed.

ObjectiveTo evaluate the effects of cardiac resynchronization therapy (CRT) on improving left ventricle (LV) function and LV asynchrony in patients with dilated cardiomyopathy(DCM)using three-dimensional ultrasound speckle tracking imaging(3D-STI).MethodsTwenty one patients with DCM and 25 normal subjects were enrolled in this study.Parameters including LV mean longitudinal peak strain (MLS),LV mean circumferential peak strain (MCS),LV mean radial peak strain (MRS),LV ejection fraction(3D-LVEF),maximal temporal difference in area strain of LV 16 segments (A-MaxTs) and its standard deviation (A-Ts-SD) were investigated by 3D-STI.LV ejection fraction (ECT-LVEF) and LV phase angle width (LVW) were obtained by radionuclide ventriculography and phase image analysis.Results The indexes of MLS,MCS,MRS and 3D-LVEF were decreased and A-MaxTs,A-Ts-SD were prolonged in DCM group compared with those of control group.There was a slightly improvement in MRS and ECTLVEF 1 week after CRT,but other Parameters including MLS,MCS,A-MaxTs,A-Ts-SD and 3D-LVEF had no significant change.Compared with corresponding values before and 1 week after CRT,all parameters (MCS,MRS,A-MaxTs,A-Ts-SD,3D-LVEF and ECT-LVEF) were further improved 6 months after CRT.MLS,MCS,MRS and 3D-LVEF had good relationship with ECT-LVEF,and A-MaxTs,A-Ts-SD had good relationship with LVW in DCM group.Conclusions3D-STI is a simple and accurate method for evaluating effects of CRT on improving LV function and LV asynchrony in patients with DCM.

ObjectiveTo explore the association between hepatic steatosis and the liver tissue expression of HBsAg and HBcAg in chronic hepatitis B (CHB) patients.MethodsFrom January 2005 to June 2008,a total of 147 CHB patients with hepatic steatosis diagnosed by liver biopsy and other 149 CHB patients without hepatic steatosis but with similar HBV DNA titer were enrolled.The differences of HBsAg and HBcAg immunostaining and liver injury in these two groups were compared.The data were analysed using t test and chi square test.ResultsCompared with non-steatosis group,the average age and body weight index of hepatic steatosis group were higher (t values were -3.31and -6.57,both P＜0.01).The percentage of moderate to severe hepatic inflammation in liver,obvious hepatic fibrosis and the strong positive HBsAg staining was lower (30.6％ vs 15.4％ ； 26.5％vs 12.8％； 23.1 ％ vs 6.7 ％； x2=9.63,8.92,15.76； all P＜0.01),and the percentage of strong positive HBcAg staining was also in downtrend.Compared with degree F1 and F2 of liver steatosis,the percentage of HBsAg and HBcAg strong positive staining in liver tissues of degree F3 and F4 was in downtrend.ConclusionsHepatic steatosis affected the expression of HBsAg and HBcAg in liver tissue of CHB patients.As hepatic steatosis appeared and became more severe,both expression of HBsAg and HBcAg and the degree of liver injury were in downtrend.

Objective To explore the possible effects of methyl methanesulfonate sensitive 2(MMS2)in the process of angiotensin Ⅱ inducing differentiation of neural stem cells (NSCs) into dopaminegic phenotype neurons. Methods NSCs were isolated from the brain of newborn rats and were cultured in the serum-free medium.Identification of neural precursor cells was done by Nestin immunocyt ochemical staining. Then the second generation of NSCs was divided into the following six groups: A, control; B, AⅡ; C, AT1 antagonist ZD7155; D, ZD7155+AⅡ; E, AT2 antagonist PD123319; F, PD123319+AⅡ. The detection of expression of MMS2 and TH mRNA level was done by real-time PCR. The silence of the expression of MMS2 in NSCs was brought about via the transfection of MMS2-siRNA, and then the NSCs were induced to differentiate into dopaminegic neurons. The expression of TH mRNA level in the cells of the groups after transfection was detected by real-time PCR. Results Nestin-positive cells were observed in suspended growth in the medium.Real-Time PCR revealed that the MMS2 and TH mRNA expression of group B and D were significantly higher than that of the control group(P<0.05), There was no significant difference in MMS2 and TH mRNA expression between group C, E, F and the control, respectively. Conclusion AⅡ increased the expression of MMS2 mRNA in NSCs and induced the differentiation of NSCs into DA neurons via AT2 recepter. MMS2 may play important roles in the process of angiotensin Ⅱ inducing NSCs to differentiate into dopaminergic neurons.