Objective: To observe the effects of knocking down the expression of peroxisome proliferators-activated receptor gamma (PPARγ) with RNA interference techniques on the proliferation and apoptosis of hepatocellular carcinoma HCCLM3 cells. Methods: A short-hairpin RNA (shRNA) eukaryotic expression vector against PPARγ was constructed and transfected into HCCLM3 cells. The changes of PPARγ expression were detected by semi-quantitative RT-PCR and Western blotting analysis. The proliferation of HCCLM3 cells was tested by MTT assay. Apoptosis ratio of HCCLM3 cells was detected by TUNEL method and flow cytometry (FCM). Expressions of PCNA and wide-type p53 protein were analyzed by immunocytochemistry (ICC) methods. Results: The sequence-specific shRNA (pshPPARγ) efficiently blocked the expression of PPARγ mRNA by 80.5%. At 48 h after transfection of pshPPARγ, proliferation of HCCLM3 cells was significantly suppressed by 71.5%. The positive rate of PCNA expression was (23.8 ± 7.2)% at 40 h transfection. The apoptotic rates were (24.2 ± 4.7)% as detected by TUNEL assay and (23.2 ± 4.2)% of cells as measured by FCM test, respectively. The detection results of the two methods were consistent. In pshPPARγ transfection group, cell cycle of HCCLM3 cells was arrested in G0/G1 phase and the proportion of cells in G2/M phase decreased. Moreover, expression of wide-type p53 protein increased significantly. Conclusion: Knockdown PPARγ expression with RNA interference technology can significantly suppress proliferation and induce apoptosis of HCCLM3 cells. It is related with up-regulation of wide-type p53 protein expression.

Objective: To observe the association of the expression of peroxisome proliferator-activated receptor γ (PPARγ) with microvessel density (MVD) and vascularization mimicry (VM) in hepatocellular carcinoma (HCC) and explore their clinical significance. Methods: Thirty-four cases of HCC confirmed by pathological examination between 2003 and 2006 in Nantong Tumor Hospital were recruited in this study. The expressions of PPARγ, MVD and VM were tested by CD34 immunohistochernical (IHC) staining and seriodic acid-schiff (PAS) reaction in 34 cases of HCC tissues and its paired non-cancerous tissues. Their biological behaviors were analyzed and associated with the clinicopathological features. Results: PPARγ was over-expressed (24/34) in HCC tissue. It was associated with tumor size (>5 cm, P =0.027), broken capsule (P =0.014),tendency to metastasis and recurrence (P =0.001). The MVD was higher (23/24) in HCC tissue than that in corresponding para-cancerous tissues (P < 0.05). High MVD was related with tumor size (>5 cm, P =0.010), broken capsule (P =0.001), tendency to metastasis and recurrence (P=0.004), with or without metastasis during follow- up period (P =0.008), and differentiation degree (P =0. 001). The positive expression of VM was detected in 7 out of 34 HCC tissues. The MVD was higher in VM-positive group (P < 0.05). The positive expressions of VM were associated with TNM stage (P =0.001), tendency to recurrence (P =0.029), and differentiation degree (P =0.004). There was a significantly positive correlation between the over-expression of PPARγ and high MVD (r = 0.519). Kaplan-Meier analysis indicated that positive expression of PPARγ and VM as well as high MVD could be used for predicting the survival rate of HCC patients (P =0.013, 0.036 and 0.022, log-rank test). Cox hazard model analysis showed that positive PPARγ expression and high MVD were the independent prognostic factors for HCC. Conclusion: The PPARγ correlates with the development of microcirculation in HCC. PPARγ is a novel prognostic indicator for evaluating the prognosis of HCC.

Vascular calcification (VC) is a chronic systemic vascular disease characterized by abnormal deposition of hydroxyapatite minerals in the vascular system and is closely associated with aging, diabetes, atherosclerosis, and chronic kidney disease. Perivascular adipose tissue (PVAT), a special type of adipose tissue that surrounds blood vessels, is thought to be a supportive component of the vascular structure and is capable of playing a role in homeostatic regulation during vasodilatation and contraction. Currently, there is growing evidence that perivascular adipose tissue acts as an endocrine and paracrine organ and interacts closely with cellular components of the vascular wall, which may be involved in the development of vascular calcification. This article reviews the role of perivascular adipose tissue in the pathophysiological process of vascular calcification and its potential as a target for therapeutic intervention, with the aim of providing new ideas for the prevention and treatment of vascular calcification.

To explore the clinical value of contrast-enhanced ultrasound (CEUS) in differentiating benign and malignant focal liver lesions (FLLs) with SonoVue, CEUS was used to examine 113 patients with focal liver lesions (FLLs) in our hospital during July 2005 to December 2006. All the patients underwent contrast-enhanced CT (CECT) or contrast-enhanced MRI (CEMRI). Except for patients with focal fatty sparings (n=18) and with hemangiomas (n=8), all the patients were confirmed by operation or ultrasonic-guided liver puncture biopsy. A sulfur hexafluoride gas-based contrast agent was used with a MI of 0.15 to 0.17. Forty-eight cases of malignant FLLs, including 30 hepatocellular carcinomas (HCCs), 2 cholangiocarcinomas and 16 metastatic tumors, were detected. Seventy-eight cases of benign FLLs, including 33 hemangiomas, 9 focal nodular hyperplasias (FNHs), 19 focal fatty sparings, 5 abscesses, 7 regenerative nodules and 2 inflammatory pseudo-tumor, were involved. The contrast pattern of benign and malignant FLLs was quite different. CEUS has higher specificity and sensitivity than conventional ultrasound in differentiating benign and malignant FLLs.
Carcinoma, Hepatocellular/*ultrasonography ; Contrast Media/*diagnostic use ; Diagnosis, Differential ; Hemangioma/ultrastructure ; Image Enhancement/*methods ; Liver Diseases/*ultrasonography ; Liver Neoplasms/*ultrastructure ; Magnetic Resonance Imaging ; Sensitivity and Specificity ; Young Adult

Objective To evaluate the relationship between transplantation of allogeneic peripheral blood stem cells (allo-PBSCs) and low incidence of leukemic relapse. Methods Thirty-seven adult patients with acute myeloid leukemia in first remission ( n =16) and chronic myeloid leukemia in chronic phase ( n =21) underwent allo-PBSCT from June 1997 to December 1999. A median number of CD34 + cells, CD3 + cells, CD4 + cells and CD8 + cells infused was 7.3 ?10 6/kg, 364?10 6/kg, 210?10 6/kg and 137?10 6/kg, respectively. Busulfan and cyclophosphamide were used as conditioning regimen (BU-CY2). All patients received cyclosporine and methotrexate for GVHD prophylaxis. Results Engraftment of neutrophil and platelet was achieved at a median of day +13(9～28) and day +12(7～19) respectively. Acute GVHD (aGVHD) occurred in 18 of 37 patients ( 48.6% ) with grade II～IV aGVHD ( 24.3% ). Chronic GVHD (cGVHD) developed in 22 (extensive 8, limited 14) out of 32 evaluable patients ( 68.8% ). 28 patients were still alive in complete remission at a median follow-up of 950 days (range 600～ 1 500 days). The main causes of death were aGVHD in two patients, CMV-IP in four patients, cGVHD in two patients, and relapse only in one patient. The probability of disease free survival at 3 years was 75.7% . Conclusion In our series, leukemia recurrence was seen only in one patient ( 2.7% ), suggesting that a high incidence of cGVHD may have an enhanced graft-versus-leukemia effect and hence reduce the incidence of relapse after transplantation.

Objective:Toprepare poly-DL-lactide-poly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 (TIMP-1) adenovirus, and to investigate their effects on the proliferation of hepatocellular carcinoma HepG2 cells. Methods: The microsphere was constructed by encapsulating recombinant adenovirus containing TIMP-1 in biodegradable PELA. The diameter of the microsphere, quantity of virus encapsulated, loading rate, and releasing kinetics were measured. HepG2 cells were infected with the microspheres; the infection efficiency was examined by fluorescent microscope; and the ultrastructure was observed by TEM. The expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR, and the proliferation of HepG2 cells was detected by MTT assay. Results: The microsphere encapsulating recombinant TIMP-1 adenovirus was successfully constructed, with its diameter, entrapment efficiency, and virus loading rate being 1.965, 60.0%, and 10.5×10~8/mg, respectively. About 60% of the viruses were released within 120 h, and the total releasing time was longer than 240 h. Infection with rAdTIMP-1 PELA microsphere efficiently induced TIMP-1 expression in HepG2 cells, and significantly inhibited the proliferation of HepG2 cells, with the inhibitory rate being 47%. Conclusion: PELA microsphere encapsulating recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for the combining macromolecular chemistry and gene therapy for treatment of hepatocellular carcinoma.

Objective To investigate the clinical skill and experience of laparoscopic herniorrhaphy in the treatment of abdominal hernia. Methods Clinical data of 30 cases of abdominal hernia undergoing laparoscopic her-niorrhaphy in Shengjing Hospital from May 2004 to December 2008 were analyzed retrospectively. There were 23 in-guinal hernia,4 incision hernia and 3 hiatal hernia. TEP in 20 cases,TAPP in 3 cases,lPOM in 4 cases and Nissen hiatal hernia repair in 3 cases were performed. Results The mean operating duration was 72 min for TEP, 127 for TAPP,150 min for IPOM,and ]60 min for hiatal hernia repair and the mean time for hospitalization was 4.6,5.8, 7.5, and 5.3 days. No conversion to open surgery happened among these cases. No recurrence was found. Conclu-sions Laparoscopic herniorrhaphy appears to be an effective and convenient technique for the treatment of abdomi-nal hernia and has many advantages of minimally invasive surgery.

Objective Off-pump coronary artery bypass grafting (OPCAB) is used more widely in recent years in China. However, there is an argument on benefits and risks of off-pump surgery. Many studies shown that OPCAB had more benefits in short-term outcomes than conventional coronary artery bypass grafting(CCABG). But evidences from other studies suggested that OPCAB resulted in less long-term graft patency as compared with on-pump surgery. This study examined the longterm graft patency of OPCAB and CCABG performed by one surgeon. Methods 50 patients who had received surgical revascularization by a surgeon for more than 5 years were reviewed, 25 patients received conventional coronary artery bypass grafting ( group 1 ) and 25 patients received OPCAB ( group 2). All patients had angiograms for compareing the graft patency between the two groups. Results Among 25 patients in group 1,21 were male and 4 were female. The mean age of patients at surgery was (55.4 ±8.9) years. 15 cases had unstable angina, 16 patients had old myocardial infarction and 6 cases had diabetes.The ejection fraction (EF) was 0.58 ±0.14. The mean number of bypasses per patient was 3.32 ±0.63. Mean duration of operation was (3.58 ± 0. 82) hours. Mean follow-up duration was ( 86.52 ± 12.48) months. 83 grafts were evaluated for patency ( open vs. closed) and were graded by Fitzgibbon as grade A ( excellent graft), B ( impaired graft, with a stenosis of ≥50%, or a diameter less than 50% of the grafted artery), or O ( completely occluded). The graft patency was 73.49%, 61grafts were graded as Fitzgibbon A, 6 grafts as Fitzgibbon B and 16 grafts as Fitzgibbon 0. 25 patients were in group 2, 21males and 4 females. The mean age of patients at procedure was (58.2 ± 9.09) years, 11 patients had unstable angina, 13 patients had old myocardial infarction and 6 cases had diabetes. The ejection fraction (EF) was 0.59 ± 0. 14. Conclusion No 2011.03.013 difference in long-term graft patency was identified between on-pump and off-pump coronary artery bypass grafting. Off-pump oronary artery bypass grafting preformed by an experienced surgeon may gain similar long-term graft patency to that of conventional bypass.

Objective To investigate the significance of cyclooxygenase-2 (COX-2),vascular endothelial growth factor(VEGF) protein expression and angiogenesis in hepatocellular carcinoma(HCC).Methods Tissue sec- tions from 40 HCC patients were examined immunohistochemically for protein expression of COX-2 and VEGF.Mi- crovessel density(MVD) was counted by endothelial cells immunostained by anti-CD34 antibody.Results The ex- pression of COX-2 protein in well differentiated HCC was stronger than that in moderately differentiated HCC and poorly differentiated HCC(P

Aim To study the anti-inflammatory activity of the Channa argus bile.Methods The bile was isolated and purified by extraction and silica gel column chromatography.Then the compounds were identified by hydrogen and carbon spectra.The spleen lymphocytes proliferation assay and Lipopolysaccharide(LPS) induced mouse macrophage RAW264.7 releasing Nitrogen Monoxide(NO) experiment were used to evaluate the anti-inflammatory activity.Results Compound(C1) of sodium taurocholate and compound(C2) of sodium taurochenodeoxycholate were isolated by activity tracing.The cell relative viabilities of the two compounds on Concanavalin A(Con A) induced spleen lymphocytes proliferation assay were 65.9%±11.7% and 60.5%±9.4%, which were significantly different from the result of model group (P<0.01), respectively.The NO production of LPS-induced RAW264.7 release of NO was (16.4±1.9) μmol·L-1 and (15.5±1.7) μmol·L-1, which were significantly different from the result of model group(P<0.01).Conclusion Sodium taurocholate and sodium taurochenodeoxycholate from Channa argus perform the anti-inflammatory activities but have no cytotoxic effect on spleen lymphocytes and macrophage.