Objective This study aimed to assess the changes in post-systolic shortening(PSS)of the left ventricular myocardium in patients with hypertrophic obstructive cardiomyopathy(HOCM)using real time three-dimensional echocardiography(RT-3DE)technology.Methods A total of 39 HOCM patients from the Department of Ultrasound,Affiliated Hospital of Jiangxi University of Chinese Medicine were included as HOCM group,and 41 healthy individuals of similar age and gender who underwent physical examinations during the same period were selected as control group.RT-3DE examinations were performed on all participants to compare conventional echocardiographic parameters,left ventricular diastolic function parameters,the distribution of PSS in different segments of the left ventricular wall,and the average post-systolic index(PSI)of the left ventricular wall.Results HOCM group exhibited significant elevations in left atrial diameter,diastolic left ventricular posterior wall thickness,left ventricular ejection fraction,left ventricular outflow tract peak pressure gradient,mitral regurgitation,and interventricular septum thickness compared to control group(P＜0.05).In contrast,left ventricular end-diastolic diameter and left ventricular end-systolic diameter were significantly decreased in HOCM group(P＜0.05).There were no significant differences in the ratio of peak early diastolic mitral inflow velocity to peak late diastolic mitral inflow velocity and the ratio of peak early diastolic mitral inflow velocity to the peak early diastolic tissue velocity of the lateral mitral annulus between two groups.However,deceleration time of mitral early diastolic inflow velocity was higher and peak late diastolic tissue velocity at mitral lateral annulus was lower in HOCM group compared to control group(P＜0.05).The probability of PSS occurring in the basal,mid,and apical segments of the interventricular septum was significantly higher in HOCM group compared to control group(P＜0.05).The average PSI in HOCM group was significantly increased compared to control group(P＜0.05).Conclusion Patients with HOCM may exhibit early myocardial shortening in the left ventricle.The application of RT-3DE technology for quantitative analysis can detect the occurrence of PSS in different segments of the left ventricle,which may have implications for clinical evaluation and intervention.

Objective This study aimed to assess the changes in post-systolic shortening(PSS)of the left ventricular myocardium in patients with hypertrophic obstructive cardiomyopathy(HOCM)using real time three-dimensional echocardiography(RT-3DE)technology.Methods A total of 39 HOCM patients from the Department of Ultrasound,Affiliated Hospital of Jiangxi University of Chinese Medicine were included as HOCM group,and 41 healthy individuals of similar age and gender who underwent physical examinations during the same period were selected as control group.RT-3DE examinations were performed on all participants to compare conventional echocardiographic parameters,left ventricular diastolic function parameters,the distribution of PSS in different segments of the left ventricular wall,and the average post-systolic index(PSI)of the left ventricular wall.Results HOCM group exhibited significant elevations in left atrial diameter,diastolic left ventricular posterior wall thickness,left ventricular ejection fraction,left ventricular outflow tract peak pressure gradient,mitral regurgitation,and interventricular septum thickness compared to control group(P＜0.05).In contrast,left ventricular end-diastolic diameter and left ventricular end-systolic diameter were significantly decreased in HOCM group(P＜0.05).There were no significant differences in the ratio of peak early diastolic mitral inflow velocity to peak late diastolic mitral inflow velocity and the ratio of peak early diastolic mitral inflow velocity to the peak early diastolic tissue velocity of the lateral mitral annulus between two groups.However,deceleration time of mitral early diastolic inflow velocity was higher and peak late diastolic tissue velocity at mitral lateral annulus was lower in HOCM group compared to control group(P＜0.05).The probability of PSS occurring in the basal,mid,and apical segments of the interventricular septum was significantly higher in HOCM group compared to control group(P＜0.05).The average PSI in HOCM group was significantly increased compared to control group(P＜0.05).Conclusion Patients with HOCM may exhibit early myocardial shortening in the left ventricle.The application of RT-3DE technology for quantitative analysis can detect the occurrence of PSS in different segments of the left ventricle,which may have implications for clinical evaluation and intervention.

Objective To investigate the effects of inhibiting epithelial mesenchymal transition (EMT) on metastastic potential of hepatocellular carcinoma (HCC) enhanced by hepatic arterial occlusion in mice. Methods Using a metastatic human HCC orthotopic nude mice model (MHCC97),the effects of hepatic artery ligation (HAL) alone,combination of HAL and phosphatidylinositol 3-kinase (PI3K)inhibitor LY294002,or combination of HAL and interferon-α (IFN-α) on the growth of planted human HCC cells and pulmonary metastasis were evaluated,respectively.The cells and tumor tissues specimens were analyzed through expression of Akt,p-Akt,E-cadhein,N-cadherin and Twist. Results HAL inhibits tumor growth (2002.97 ± 331.28 ) mm3 vs.( 3921.23 ± 786.21 ) mm3,t =4.052,P ＜ 0.01 ),while promoting pulmonary metastatsis ( 10/12 vs.4/12,P ＜ 0.05).HAL combined with LY294002 represses significantly enhanced pulmonary metastasis rate by HAL alone (0/6 vs.6/6,P ＜ 0.01 ).Moderate-dose IFN-α (7.5 × 106 U/kg) combined with HAL failed to further reduce tumor volume compared with that of HAL alone,but inhibited markedly pulmonary metastasis (2/6 vs.6/6,P ＜ 0.05 ).The augmented level of N-cadherin and Twist in tumor tissues due to HAL reduced by LY294002 or 7.5 × 106 U/kg IFN-α.The arrest of EMT by LY294002 or IFN-α in HAL-treated xenografts was further demonstrated by the in vitro response of hypoxic cells to both agents. Conclusions Inhibition of EMT in HCC cells could repress enhanced metastastic potential due to hepatic arterial occlusion.

ObjectiveTo explore the role of β-catenin in the proinvasive consequences of hypoxia in hepatocellular carcinoma (HCC).MethodsWe established in vitro and in vivo hypoxic models using the highly metastatic MHCC97 and the stable red fluorescent protein-expressing MHCC97-R cells.The role of β-catenin in hypoxia-mediated aggressiveness was investigated by β-catenin knockdown.ResultsHypoxia caused a pronounced arrest of proliferation in MHCC97 cells,suppressed tumor growth in MHCC97-R xenografts,but promoted in vitro invasiveness and in vivo metastasis.β-Catenin-silencing by short hairpin significantly inhibited the enhanced invasiveness of MHCC97 cells due to hypoxia,reduced the increase in distant metastasis by hepatic arterial ligation,but failed to further restrain cell proliferation.Conclusionβ-Catenin in HCC cells plays an essential role in the hypoxia-induced metastatic potential.A reduction of βcatenin expression inhibited the proinvasive consequences of hypoxia in HCC.

Objective To investigate the effects of Pseudomonas aeruginosa vaccine (PA) on proliferation and invasiveness of the hepatocellular carcinoma cell line MHCC97L with metastatic potential. Methods Proliferation, growth curve, plate efficiency, flow cytometry, transwell invasion assay, cell motility assay, scarification test, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP2) protein activity were evaluated after cells were treated with PA at various concentrations. Results PA can inhibit the proliferation and plate efficiency of MHCC97L cell markedly in a dose-dependent manner. The IC50 of cells treated with PA for 48 h and 72 h was 3.1 ×108/ml and 1.9 × 108/ml, respectively. The doubling time increased and plate efficiency decreased gradually when cells treated with 0.5 × 108/ml, 1 × 108/ml and 2 × 108/ml PA (P＜0.01). PA could induce cell cycle arrest at the G1 phase in a dose-dependent manner by flow cytometric analysis. The average amount of invading cell per field in cell invasion assay and motility assay were 4. 8 ± 1.3 and 8. 8±2.2 when cells treated with 1× 108/ml PA, which was significantly lower than that of control group (8. 6±2. 1 and 15. 6±1.2 ) (P＜0.01) Scarification test showed that the metastatic ability of cells treated with 1 × 108/ml PA significantly lower than that in the control group. Comparison between cells treated with 1 × 108/ml PA and control group, no remarkable difference was found regarding expression of VEGF and MMP2 in supernatant of cell culture. Conclusion PA can inhibit proliferation and plate efficiency of HCC cell line MHCC97L, which is in part mediated by the cell cycle arrest at the G1 phase. PA could inhibit invasiveness of HCC cell line MHCC97L, which is unrelated to the VEGF and MMP2 protein activity.

Objective To investigate the effects of hypoxia on malignant phenotype of a hepatocellular carcinoma(HCC)cell line and its molecular basis.Methods Human HCC cell line with highly metastatic potential(MHCC97H)was grown under hypoxia(induced by 100 μmol/L CoCl_2)and normoxic conditions respectively.To observe the effects of hypoxia on MHCC97H cells was observed,the tumorigenicity was carried out by soft agar cloning method in vitro and inoculation in nude mice in vivo;Invasive and metastatic potential were measured.Experiments with and/or without Matrigel in vitro and a metastatic human HCC orthotopic nude mice model by HAL in vivo.To clarify the molecular background,the proliferation of cells were analyzed by MTT assay and the cell cycle was detected by flow cytometry;The expression of embryonic stem cell(ES)-like genes(Oct4,Nanog and Sox2)were detected by real-time RT-PCR;CD90~+ and CD133~+ subpopulation from MHCC97H cells were isolated by flow cytometry and the expressions of CD90 and CD133 of MHCC97H cells were analyzed by immunofluorescence.Results Hypoxia increased tumor migration(t=2.792,P=0.023),invasiveness(t=7.624,P＜0.0001)and clone forming ability(t=3.292,P=0.011)of MHCC97H cells in vitro,and promoted tumorigenesis(x~2=8.571,P=0.015)and pulmonary metastasis(x~2=5.507,P=0.031)in vivo.The proliferation of MHCC97H cells arrest under hypoxic conditions accompanied with increased proportion of cells in G1 phase,the expressions of Oct4,Nanog,Sox2 significantly increased in response to hypoxia,but the fluorescence intensity and number of CD90~+ and CD133~+ MHCC97H cells decreased or unchanged.Conclusion Hypoxia increases aggressiveness of MHCC97H cells,which was correlated with the acquisition of embryonic stem cell-like characteristics.

OBJECTIVEThe goal of this study is to investigate the inappropriate activation of Wnt pathway in the hepatocarcinogenesis.

METHODSWe analyzed the alterations of three key components of Wnt pathway, beta-catenin, glycogen synthase kinase 3beta (GSK-3beta) and T cell factor 4 (Tcf-4), in 34 samples of hepatocellular carcinoma (HCC) and paracancerous normal liver by immunohistochemistry, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), direct sequencing, semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization.

RESULTSWe found 61.8% (21/34) of all the HCCs examined showed an abnormal beta-catenin protein accumulation in the cytoplasm or nuclei. RT-PCR-SSCP and direct sequencing showed that beta-catenin exon 3 mutations existed in 44.1% (15/34) of the HCCs. No mutations of GSK-3beta or Tcf-4 were detected in HCCs. Moreover, mRNA of beta-catenin and Tcf-4 but not GSK-3beta was found to be over expressed in HCCs. On analyzing the relationship between alterations of beta-catenin or Tcf-4 and C-myc or Cyclin D1 expression, we found that the mutations of beta-catenin as well as over expression of beta-catenin or Tcf-4 gene were independently correlated with C-myc gene over expression in HCCs.

CONCLUSIONSOur present findings strongly suggest mutations of beta-catenin as well as over expression of beta-catenin and Tcf-4 gene activate the Wnt pathway in HCC independently with the target gene most likely to be C-myc.

Carcinoma, Hepatocellular ; metabolism ; Cytoskeletal Proteins ; genetics ; physiology ; Glycogen Synthase Kinase 3 ; genetics ; physiology ; Glycogen Synthase Kinase 3 beta ; Humans ; Immunohistochemistry ; Liver Neoplasms ; metabolism ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Proto-Oncogene Proteins ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; TCF Transcription Factors ; Trans-Activators ; genetics ; physiology ; Transcription Factor 7-Like 2 Protein ; Transcription Factors ; genetics ; physiology ; Wnt Proteins ; Zebrafish Proteins ; beta Catenin

OBJECTIVETo study the immunoprotective effect of IL-2 and B7-1 gene co-transfected liver cancer vaccine on hepatocarcinogenesis in mice.

METHODSThe murine liver cancer cell strain Hepal-6 was transfected with IL-2 and/or B7-1 gene via recombinant adenovirus vectors and the liver cancer vaccines were prepared. C57BL/6 mice were immunized with the vaccines and challenged with the parental Hepal-6 cells afterwards. The immunoprotection was investigated and the reactive T cell line was assayed.

RESULTSThe effect of the Hep6-IL2/B7 vaccine on the onset of tumor formation was the strongest. The media survival time of the mice was the longest (68 days, P<0.05) and the implanted tumor was the smallest (P<0.05). The effect of single IL-2 or B7-1 gene-transfected vaccine was next to the co-transfected gene group with the mean survival time being 59 and 54 days, respectively. The mean survival time of wild or BGFP gene modified vaccine immunized group was 51 and 48 days, respectively. The control group all died within 38 days and the implanted tumor was the largest (P<0.05). The cellular immunofunction test and cytotoxicity study showed that the mice immunized with the Hep6-IL2/B7 vaccine gained significantly increased NK, LAK and CTL activity (29.0% +/- 2.5%, 65.0% +/- 2.9%, 83.1% +/- 1.5% respectively, compared with other groups P<0.05).

CONCLUSIONSThe IL-2 and B7-1 gene co-transfected liver cancer vaccines can induce the mice to produce activated and specific CTL against the parental tumor cells, and demonstrate stronger effect on the hepatocarcinogenesis than single gene modified or the regular tumor vaccine. Therefore, the vaccines may become a novel potential therapy for recurrence and metastases of HCC.

Adenoviridae ; genetics ; Animals ; B7-1 Antigen ; genetics ; immunology ; Cancer Vaccines ; administration & dosage ; genetics ; immunology ; Cell Division ; immunology ; Female ; Genetic Vectors ; administration & dosage ; genetics ; immunology ; Humans ; Interleukin-2 ; genetics ; immunology ; Killer Cells, Lymphokine-Activated ; immunology ; Killer Cells, Natural ; immunology ; Liver Neoplasms, Experimental ; immunology ; pathology ; prevention & control ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Survival Analysis ; T-Lymphocytes, Cytotoxic ; immunology ; Time Factors ; Transfection ; Tumor Cells, Cultured

OBJECTIVETo compare gene expression profile of human hepatocellular carcinoma (HCC) cell lines with different metastatic potentials, so as to screen for metastasis-related genes.

METHODSGene expression profile of MHCC97-L and HCCLM3, two HCC cell lines with similar genetic background but different in spontaneous metastatic potentials, were studied by cDNA microarray.

RESULTSFrom 1,626 screened genes, 25 differentially expressed genes were found, 18 showed decreased expression including the decreased expression of cell cycle control genes Rb2, mismatch repair gene hMSH2, and signal transduction gene protein kinase C beta 2 and 7 increased expression including signal transduction gene MAP kinase kinase 6, cell proliferation gene E25, immunity related gene SP40, 40, etc in HCCLM3.

CONCLUSIONThe genes, being closely associated with cancer metastasis, could be considered as potential markers to predict metastasis and targets for anti-metastasis intervention.

Blotting, Northern ; Carcinoma, Hepatocellular ; genetics ; pathology ; Gene Expression ; Gene Expression Profiling ; Humans ; Liver Neoplasms ; genetics ; pathology ; Neoplasm Metastasis ; genetics ; Oligonucleotide Array Sequence Analysis ; RNA, Neoplasm ; analysis ; Tumor Cells, Cultured

Objective To make a better understanding of the molecular mechanisms involved in recurrence and metastasis of the hepatocellular carcinoma (HCC), some invasion related oncogenes, and growth factors have been investigated. Methods The studies were seperately carried out, the results of which were summarized in this article with relation to tumor size and invasiveness of HCC.Results The aberration rates of p53 and CDKN2 in HCC were 45.9% and 36.4% respectively, which were higher in invasive HCC compared with non-invasive HCC. H-ras expression was positive in 29.3% of HCC, which was associated with recurrence and extrahepatic metastasis of HCC. Intralesional injection of H-ras antisense gene markedly inhibited the tumor growth and metastasis of HCC in nude mice. The positive rates of transforming growth factor (TGF)-alpha, epidermal growth factor receptor (EGFR) and c-erbB-2 were 45.7%, 47.1% and 92.3% respectively. The expression of EGFR was closely related to TGF-alpha, which was related to HCC recurrence. But no obvious difference of TGF-alpha or c-erbB-2 expression was found between HCC with and without recurrence, or with and without extrahepatic metastasis. Expression of nm23 / tissue inhibitor of metalloproteinase (TIMP)-2 was positively associated with the prognosis of HCC patients (Log-rank, P<0.001). The alterative rates of above-mentioned genes and growth factors in small HCC were slightly lower than that in large ones, but no significant difference was shown except the p53 mutation.Conclusions The p53/CDKN2 mutation, overexpression of H-ras/EGFR, were associated with the invasiveness and recurrence of HCC. H-ras antisense gene might be of potential implication in the control of HCC recurrence and metastasis. Expression of nm23/TIMP-2 was closely related to the prognosis of HCC patients. Biological characteristics remained critical points to the prognosis even in small HCC.