Overexpression of UBE2C correlates with poor prognosis in gastric cancer patients.

OBJECTIVE: The ubiquitin-conjugating enzyme E2C (UBE2C) has been known as a crucial factor upregulated in various tumors. The functions of UBE2C is mainly involved in the pathway protein ubiquitination. This study investigates the expression of UBE2C in gastric cancers and its correlation with overall survival rate. MATERIALS AND METHODS: Real-time PCR (RT-PCR) and Western blotting were performed to determine the expression of UBE2C in gastric cancer samples and adjacent normal tissues. Immunohistochemical staining was used to assess the expression of UBE2C in 216 paraffin-embedded gastric cancer tissues. RESULTS: The mRNA and relevant protein levels of UBE2C in gastric cancer tissues are significantly greater than those in the adjacent normal tissues. Also, the expression of UBE2C is found to correlate with lymphatic metastasis, serosa invasion, TNM (Malignant Tumors) staging and Lauren's classification (p<0.05). The univariate analysis shows that the overexpression of UBE2C associates with poor prognosis (p=0.001). The multivariate analysis demonstrates that expression of UBE2C, lymphatic metastasis, and TNM staging are independent prognostic indicators. CONCLUSIONS: This study shows that overexpression of UBE2C contributes to the development of gastric cancer, and UBE2C has the potential to be exploited as a therapeutic target.

Can catheter-directed thrombolysis be applied to acute lower extremity artery embolism after recent cerebral embolism from atrial fibrillation?

PURPOSE: To assess the feasibility and efficacy of catheter-directed thrombolysis with recombinant tissue plasminogen activator (rt-PA) for acute limb embolism in patients with recent cerebral embolism due to atrial fibrillation. MATERIALS AND METHODS: Eight patients (six men, two women; mean age 63.5 years) with acute embolic occlusion of two left common iliac arteries, four femoral arteries (three left; one right), and two right popliteal arteries were treated. All patients had a history of recent cerebral embolism (mean 6 days, range 5-15 days) and all had a history of atrial fibrillation (duration 5-10 years). Catheter-directed thrombolysis started a few hours (mean 6.2h; range 3-10h) after the onset of arterial embolism. Two 5mg boluses of rt-PA were injected into the proximal clot through a 5 F end-hole catheter and, subsequently, two additional boluses of 5mg rt-PA were injected into the emboli. In patients with residual emboli, infusion with rt-PA (1mg/h) was continued. Percutaneous transluminal angioplasty was performed in three patients, and a stent was deployed in one patient. RESULTS: Technical success was achieved in all patients. Clinical success rate was 87.5% (7/8). The one clinical failure was secondary to chronic occlusion of outflow runoff vessels. The mean duration of continuous rt-PA infusion was 3.6h, the mean total dose of rt-PA administered was 23.6 mg (range 20-28 mg). There was no significant change in stroke scale scores during thrombolysis and no intracerebral haemorrhage was found at computed tomography (CT) after thrombolysis. Minor complications included haematomata at puncture sites (6/8), bleeding around the vascular sheath (2/8), and haematuria (1/8). During the follow-up period of 3-6 months, one patient suffered from recurrent cerebral embolism and died. CONCLUSIONS: Catheter-directed thrombolysis with rt-PA is an option for acute lower extremity arterial embolism in patients with recent cerebral embolism and a history of atrial fibrillation. Further studies should be undertaken to determine the risk of intracerebral haemorrhage caused by catheter-directed thrombolysis in individual stroke patients.

Beneficial effects of jejunal continuity and duodenal food passage after total gastrectomy: a retrospective study of 704 patients.

AIM: To evaluate effects of reconstruction procedures on post-operative outcomes and nutritional status after total gastrectomy. METHODS: The study group comprised 704 consecutive patients with gastric cancer who underwent total gastrectomy between December 1985 and December 2003. Six alimentary reconstruction procedures were performed, including jejunal continuity [Braun, modified Braun I and II and functional jejunal interposition (FJI)] and jejunum transection ["P" Roux-en-Y and "P" jejunal interposition (PJI)]. The duodenal food passage was maintained only by FJI and PJI. We evaluated the time interval to restore food intake after surgery and the incidence of complications and nutritional status for 12 months. RESULTS: Patients who received jejunum transection required 7.8+/-2.5 days and 11.9+/-4.9 days to restore liquid and semi-liquid food intake, respectively, which reduced to 3.9+/-2.1 days for liquid and 7.9+/-3.9 days for semi-liquid food intake by jejunum continuity. The incidence rates of reflux esophagitis and Roux-en-Y syndrome in patients receiving jejunum transection were 23.5% and 42.4%, respectively, which were decreased to 9.35% and 14.7%, respectively, by jejunal continuity. Furthermore, prognostic nutrition index score of patients receiving the procedures maintaining duodenal food passage (52.9+/-10.9) was higher than that of patients without the duodenal food passage (46.7+/-8.2). CONCLUSION: Jejunal continuity and duodenal food passage showed beneficial effects on clinical outcomes after surgery. Among these six procedures, FJI was the only procedure to combine the benefits of jejunal continuity and maintaining the duodenal food passage, indicating that FJI has potential clinical application to improve the quality of patient's life after total gastrectomy.

Up-regulation of Twist induces angiogenesis and correlates with metastasis in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a highly invasive tumor characterized by vigorous neovascularization. The purpose of this study is to examine the expression of Twist, a highly conserved bHLH transcription factor that is known to promote EMT, and evaluate its effect on tumor angiogenesis and metastasis of HCC. The mRNA expression of Twist, VEGF, E-cadherin, and N-cadherin was determined by Real-Time RT-PCR in 30 pairs of hepatocellular carcinomas and matched non-cancerous tissues. Immunohistochemistry was carried out to analyze the protein expression of Twist, VEGF, E-cadherin, and N-cadherin in 40 hepatocellular carcinoma cases. The staining of endothelial cells for CD34 was used to evaluate the MVD. We found that Twist mRNA and protein were both increased in HCC as compared to non-cancerous tissues. The HCC specimens showing positive Twist expression had a higher microvessel density than those without Twist expression. And up-regulated Twist protein was significantly associated with intrahepatic and extrahepatic metastasis (p=0.048 and P=0.039 respectively). In addition, patients with Twist expression had poor prognosis. We also found that the expression of Twist positively correlated with up-regulation of VEGF and N-cadherin (P=0.002 and p=0.016 respectively), but not with downregulation of E-cadherin in HCC. Our results demonstrate that Twist may play an important role in the angiogenesis and metastasis of HCC. Twist expression may become a potential novel prognostic factor for the disease survival of HCC.

Reversing adriamycin resistance of human breast cancer cells by hyperthermia combined with Interferon alpha and Verapamil.

One of the major obstacles related to chemotherapy is resistance against anticancer drugs, including Adriamycin (ADM). The purpose of the present work is to investigate the reversal effects on ADM resistance by hyperthermia (42.5 degrees C) combined with two reversal agents (Interferon alpha and Verapamil) in MCF-7/ADR (ADM-resistant MCF-7 breast cancer cell line), and its relevant molecular mechanism of action. The cell survival rate and ADM IC50 of different experiment groups were measured by MTT test. The quantitative expression of MDR1 gene in cells was detected by Real-time PCR, and the expression of P-glycoprotein (P-gp) on the cells surface and the intracellular ADM accumulation was detected by flow cytometry (FCM). The ADM IC50 of the MCF-7/ADR cells decreased 830-fold after combined with Interferon alpha (IFN-alpha) and Verapamil (VRP). Although there was no distinction in the mRNA expression of MDR1, the P-gp on the MCF-7/ADR cell membrane was significantly reduced and the cellular ADM uptake increased markedly as compared to pretreatment. Our results suggeste that hyperthermia induces a considerably reversal activity against ADM resistance synergizing other reversal agents (IFN-alpha and VRP). The reversal mechanism needs further study. However, these features of hyperthermia may be exploited in clinical cancer chemotherapy.

Correlation of four vascular specific growth factors with carcinogenesis and portal vein tumor thrombus formation in human hepatocellular carcinoma.

The aim of the present study was to detect the correlation between the expression of vascular endothelial growth factor (VEGF), angiopoietin 2 (Ang2), ephrinB2 and endocrine gland-derived vascular endothelial growth factor (EG-VEGF) and carcinogenesis or portal vein tumor thrombus (PVTT) formation in human hepatocellular carcinoma (HCC). The expression of VEGF, Ang2, ephrinB2 and EG-VEGF was detected by RT-PCR in 54 cases HCC without PVTT (group A), 9 cases HCC with PVTT (group B), 10 normal liver tissues (group D) and 10 cirrhosis tissues (group C). The samples were also stained with CD34 by immunohistochemistry. Quantitation of microvessel density (MVD) and semi-quantitation of VEGF, Ang2, ephrinB2 and EG-VEGF expression were analyzed to find the relations. The MVD was 146.69 +/- 77.79, 214.07 +/- 54.41, 32.85 +/- 8.49 and 34.83 +/- 8.29 in group A-D respectively with significant difference (F = 19.77, P = 0.000). The MVD in group A was higher than that in group C P = 0.006, but lower than that in group B P < or = 0.05 or 0.01. The expression levels of VEGF165, VEGF189, Ang2 and EG-VEGF mRNA were significantly different among the groups. The expression levels of VEGF165, Ang2 and EG-VEGF mRNA in group A were all higher than those in group C, but lower than those in group B P < 0.05 or 0.01. The MVD was significantly correlated with VEGF165, VEGF189, Ang2 and EG-VEGF mRNA with Spearman's related coefficient being 0.764, 0.510, 0.640 and 0.366 in HCC (P = 0.000, 0.000 0.000 and 0.003). In conclusion VEGF, Ang2 and EG-VEGF mRNA may play a role in angiogenesis and carcinogenesis of HCC. They can promote PVTT formation in HCC by modulating angiogenesis.

Efficacy of postoperative transarterial chemoembolization and portal vein chemotherapy for patients with hepatocellular carcinoma complicated by portal vein tumor thrombosis--a randomized study.

OBJECTIVES: The aim of this single, randomized study was to explore the efficacy of postoperative transarterial chemoembolization (TACE) and portal vein chemotherapy (PVC) for patients with hepatocellular carcinoma (HCC) complicated by portal vein tumor thrombosis (PVTT) and to evaluate prognostic factors. METHODS: The study cohort consisted of 112 patients with HCC and PVTT randomly divided into three groups: Group A (37 patients), operation only; Group B (35 patients), operation plus TACE; Group C (40 patients), operation plus TACE and PVC. Disease-free survival rates and prognostic factors were analyzed. RESULTS: Most of the side effects and complications were related to the operation, catheters, and local chemotherapy and included liver decompensation (15.0%), catheter obstruction (11.6%), and nausea and loss of appetite (22.1%). The disease-free survival curve was significantly different among the three groups, as estimated by the Kaplan-Meier method (both P < 0.05). Group C showed a significantly higher disease-free survival rate than Group A (P < 0.05), but no statistical differences were found between group A and group B, and group B and group C (both P > 0.05). Tumor size, tumor number, PVTT location, and treatment modalities were independent prognostic factors (P < 0.05). CONCLUSION: Postoperative TACE combined with PVC may benefit the survival of patients with HCC complicated by PVTT in the short-term (less than 60 months), but long-term efficacy is not yet certain and needs to be confirmed by further studies.

Postoperative transhepatic arterial chemoembolization and portal vein chemotherapy for patients with hepatocellular carcinoma: a randomized study with 131 cases.

OBJECTIVE: To explore the value of postoperative transarterial chemoembolization (TACE) and portal vein chemotherapy (PVC) in patients with hepatocellular carcinoma (HCC) in the prevention of recurrence, and to evaluate prognostic factors in a randomized study. METHOD: 131 patients with HCC were randomly divided into 3 groups: operation only (group A, n = 45); operation plus TACE (group B, n = 39), and operation plus TACE and PVC (group C, n = 47). Disease-free survival rates as well as prognostic factors were analyzed. RESULTS: Most of the side effects and complications related to the operation, catheters and local chemotherapy were liver decompensation (16.1%), catheter obstruction (12.9%), and nausea and loss of appetite (25.8%), respectively. The disease-free survival curves were significantly different between the 3 groups as estimated by the Kaplan-Meier method (p < 0.05). Group C had a significantly higher disease-free survival rate compared to group A (p < 0.05). But no statistical differences were found between groups A and B and groups B and C (both p > 0.05). Tumor number and treatment modalities were independent prognostic factors for HCC patients (p < 0.05). CONCLUSION: Postoperative TACE combined with PVC may benefit the survival of patients with HCC. In specialized medical centers, aggressive methods such as TACE and PVC should be attempted on HCC patients without contraindications.

High-resolution cDNA microarray CGH mapping of genomic imbalances in osteosarcoma using formalin-fixed paraffin-embedded tissue.

Formalin-fixed paraffin embedded (FFPE) tumor tissue provides an opportunity to perform retrospective genomic studies of tumors in which chromosomal imbalances are strongly associated with oncogenesis. The application of comparative genomic hybridization (CGH) has led to the rapid accumulation of cytogenetic information on osteosarcoma (OS); however, the limited resolving power of metaphase CGH does not permit precise mapping of imbalances. Array CGH allows quantitative detection and more precise delineation of copy number aberrations in tumors. Unfortunately the high cost and lower density of BACs on available commercial arrays has limited the ability to comprehensively profile copy number changes in tumors such as OS that are recurrently subject to genomic imbalance. In this study a cDNA/EST microarray including 18,980 human cDNAs (which represent all 22 pairs of autosomal chromosomes and chromosome X) was used for CGH analysis of eight OS FFPE. Chromosomes 1, 12, 17, and X harbored the most imbalances. Gain/amplification of X was observed in 4/8 OS, and in keeping with other recent genomic analyses of OS, gain/amplification of 17p11.2 was often accompanied by a distal deletion in the region of the p53 gene. Gain/amplification of the X chromosome was verified using interphase FISH carried out on a subset of OS FFPE sections and OS tissue arrays.

Twenty-year trends of primary liver cancer incidence rates in an urban Chinese population.

The objective of this study was to describe trends in the incidence rates of primary liver cancer in a geographically defined Chinese population. Primary liver cancer cases (N=13 685) were diagnosed between 1981 and 2000 and identified by the Tianjin Cancer Registry. Age-adjusted and age-specific incidence rates were examined in both males and females. Poisson regression was employed to assess the incidence rate trends. Crude and age-adjusted incidence rates in the study period were: 27.4/100 000 and 16.4/100 000 in males and 11.5/100 000 and 6.4/100 000 in females, respectively. While the results from Poisson regression analyses suggest statistically significant trends of declining incidence rates of primary liver cancer overall, trends were not consistent across age and sex groups. The decline in incidence was observed, for the most part, in the 40-69 age group, with a greater decrease in males. Our findings provide a new evidence of a downward trend in incidence rates of this disease in China for a period of 20 years. As the observed decline is relatively small and inconsistent across sex and age groups, a continued epidemiological observation on this condition is required.

Potential mechanisms of leukemia cell resistance to TRAIL-induced apopotosis.

There are many factors contributing to the resistance to TRAIL (Tumor necrosis factor-related apoptosis-inducing ligand)-induced apoptosis. However, it is not clear whether the mechanism of resistance to TRAIL is constitutive or inductive. Therefore, the purpose of this study was to investigate the resistant mechanisms to TRAIL at different levels in the apoptotic pathway. The human T-lymphoblastic leukemic CEM cell line showed more resistant to TRAIL-induced apoptosis compared with the human chronic myeloid leukemic K562 cell line. Lower level of constitutive caspase-8 expression in the CEM cell line led to a poor response to both TRAIL-induced activation of caspase-3 and reduction in the mitochondrial membrane potential (DeltaPsim). There was no significant difference in the constitutive levels of NF-kappaB in CEM and K562 cell lines. However, CEM cells showed a faster response to TRAIL-induced NF-kappaB activation than K562 cells. TRAIL-induced regulation of Bcl-2 family of proteins included an up-regulation in Bcl-2/Bcl-XL and a down-regulation in Bax. IAPs, such as XIAP, cIAP-1, cIAP-2 and Survivin were all up-regulated during the treatment with TRAIL. In summary, our data suggest that the leukemic cells resistance to TRAIL-induced apoptosis might be due to the deficiency in the constitutive caspase-8 expression. Development of potential resistance to apoptosis by TRAIL can occur in both TRAIL-resistant and TRAIL-sensitive leukemic cells.

Small bowel metastases of malignant melanoma: palliative effect of surgical resection.

Malignant melanoma shows an unusual predilection to metastasize to the small intestine. Three patients with malignant melanoma involving the small bowel are reported. One patient was operated on for small bowel obstruction and the other two for gastrointestinal bleeding. Two patients remained well 6 and 2 years, respectively, after surgery. One patient died of metastatic melanoma 4 years post-operation. Metastatic melanoma in the small bowel should be suspected in any patient with a previous history of malignant melanoma who develops GI symptoms or chronic blood loss. Surgical treatment was the first choice; the prognosis after surgical resection was much better than for other organ metastases or simultaneous metastases of the small bowel and other organs.

Determination of human T cell activity in response to allogeneic cells and mitogens. An immunochemical assay for gamma-interferon is more sensitive and specific than a proliferation assay.

T lymphocytes proliferate and secrete lymphokines in response to allogeneic cells, mitogens and other stimuli. Cell proliferation as measured by [3H]thymidine ([3H]Tdr) incorporation into DNA has been routinely used to determine T cell responses in research and clinical laboratories. We have compared the sensitivity of an immunoradiometric assay (IRMA) for human gamma-interferon (IFN-gamma) (Chang et al., 1984), with that of the conventional [3H]Tdr incorporation assay in the measurement of T cell responses to antigens and mitogens in culture. Peripheral blood mononuclear cells (PBMs) were incubated in the presence and absence of phytohemagglutinin (PHA) or mononuclear cells from another individual for various periods of time. The culture fluids were collected for determining IFN-gamma and the cells were assayed for [3H]Tdr incorporation. Results of measurements were expressed in terms of stimulation indices. Both IFN-gamma secretion and thymidine incorporation were measurable in mixed lymphocyte cultures after incubation for 3 days, and in PHA stimulated culture after 24 h of incubation. The stimulation indices reflecting increased gamma-interferon were found to be more pronounced and more consistent than those of [3H]Tdr incorporation.