Effects of Epstein-Barr Virus on Proliferation，Apoptosis and Lipid Metabolism of Gastric Cancer Cells / 中山大学学报(医学科学版)

@# 【Objective】To investigate the effects of Epstein-Barr virus（EBV）on proliferation，apoptosis and lipid metabolism of gastric cancer cells，revealing the pathogenesis and development of EBV-associated gastric carcinoma （EBVaGC）.【Methods】EBV-positive cell line AGS-EBV was established by co-culturing AGS and Akata. Then we compared proliferation，apoptosis and lipid metabolism level between AGS and AGS-EBV cells. CCK-8 assays and Annexin V PE/7-AAD assays were performed to determine the proliferation and apoptosis of gastric cancer cells. Oil-Red O staining and three kinds of kits were used to detect lipid contents including lipid droplets，free fatty acid，triacylglycerol and total cholesterol. Key enzymes of lipogenesis were measured by qRT-PCR.【Results】EBV promoted the proliferation of gastric cancer cell line AGS（F = 23.214，P = 0.001；P values of 24 h，48 h，72 h，96 h and 120 h were 0.007， 0.004，＜0.001，＜0.001 and ＜0.001，respectively），inhibited apoptosis（P values of late and total apoptotic rates were 0.032，< 0.001），and increased intracellular lipid droplets，free fatty acids（P < 0.001），triacylglycerol（P = 0.004） and total cholesterol（P < 0.001）probably via lipogenesis. 【Conclusions】EBV promotes proliferation，inhibits apoptosis， and enhances lipid metabolism of gastric cancer cells.

Prognostic power of abnormal cytogenetics for multiple myeloma: a multicenter study in China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Chromosomal abnormalities have been shown to play an important prognostic role in multiple myeloma (MM). Interphase fluorescence in situ hybridization (i-FISH) has been much more effective to identify cytogenetic aberrations in MM than conventional cytogenetic technique (CC). To clearly determine the cytogenetic features of Chinese MM patients and identify their prognostic implications, we designed a multicenter study based on i-FISH including 672 patients from 52 hospitals in China.</p><p><b>METHODS</b>All 672 patients were systematically screened for the following genomic aberrations: del(13q), IgH rearrangement, del(p53) and 1q21 amplifications.</p><p><b>RESULTS</b>The analysis showed that the chromosomal changes were detected in 22.1% patients by CC and in 82.3% patients by i-FISH. The most common abnormalities by CC were chromosome 1 aberrations (48.4%), -13/13q- (37.6%), hyperdiploidy (36.6%), hypodiploidy (30.1%) and IgH rearrangements (23.7%). The most frequent abnormalities by FISH was del(13q), which was found in 60.4% patients, whereas IgH rearrangement, 1q21 amplification and p53 deletions were detected in 57.6%, 49.0% and 34.7% cases, respectively. By statistical analysis, -13/13q- by CC was associated with low level of platelet (P = 0.015), hyperdiploidy was associated with low level of serum albumin (P = 0.028), and IgH rearrangement by FISH was associated with high level of β2 microglobulin (P = 0.019). Moreover, 1q21 amplification and del(p53) by FISH conferred a high incidence of progressive disease (PD) after initial therapy. Metaphase detection of IgH rearrangements and chromosome 1 aberrations concurrently was associated with a short progression free survival (PFS) (P = 0.036). No significant prognostic implications of other cytogenetic abnormalities were found associated with overall survival and PFS.</p><p><b>CONCLUSIONS</b>Chinese MM patients had similar cytogenetic abnormalities compared with the previous reported studies. However, the prognostic significance of FISH aberrations were not clearly determined and further study is required.</p>

Correlation of Fas/FasL expression to cell apoptosis in Epstein-Barr virus-associated gastric carcinoma / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Epstein-Barr virus (EBV) has been detected in about 10% of gastric carcinomas. However, the pathogenetic role of EBV in gastric carcinoma is uncertain. This study was to explore the correlation of Fas/FasL expression to the apoptosis of tumor cells and tumor-infiltrating lymphocytes (TIL) in EBV-associated gastric carcinoma (EBVaGC).</p><p><b>METHODS</b>Fas/FasL expression in 49 specimens of EBVaGC, 20 specimens of EBV-negative gastric carcinoma (EBVnGC) and 12 specimens of normal gastric mucosa was detected by immunohistochemistry. The apoptotic index (AI) of cells was determined by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL).</p><p><b>RESULTS</b>The positive rates of Fas were 91.7% in normal gastric mucosa and 76.8% in gastric carcinoma (P < 0.05); those of FasL were 16.7% in normal gastric mucosa and 58% in gastric carcinoma (P < 0.05). The positive rate of Fas was significantly lower in EBVaGC than in EBVnGC (71.4% vs. 90.0%, P < 0.05). The positive rate of FasL in EBVaGC was significantly higher than that in EBVnGC (63.2% vs. 45%, P < 0.05). The AI of EBVaGC cells was significantly lower than that of EBVnGC cells (P = 0.002). The number and AI of TIL in EBVaGC were higher than those in EBVnGC (P < 0.05). The AI of TIL was positively correlated with the level of FasL expression in tumor cells (r=0.237, P = 0.028).</p><p><b>CONCLUSION</b>Up-regulation of FasL expression and decrease of TIL apoptosis in EBVaGC may facilitate the escape of tumor cells from the host immunosurveillance, and it might contribute to the development and progression of carcinoma.</p>

Clinicopathologic features and protein expression study of Epstein-Barr virus-associated gastric carcinoma in Guangzhou / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the prevalence of Epstein-Barr virus (EBV)-associated gastric carcinomas in Guangzhou, their clinicopathologic features and related protein expressions including DNMT1, p16, and cyclin D1.</p><p><b>METHOD</b>A total of 676 cases of EBV-associated gastric carcinoma were included in the study. The presence of EBV-encoded small RNA1 (EBER1), a marker for EBV infection, was analyzed by in-situ hybridization using formalin-fixed and paraffin-embedded tumor samples. Expression of EBV-encoded proteins, DNMT1, p16 and cyclin D1 were detected by immunohistochemistry.</p><p><b>RESULTS</b>Forty-five of 676 gastric carcinomas showed EBER intranuclear positivity in all tumor cells. EBV involvement was significantly more frequent among the male than the female patients, especially in tumors of less differentiated types (diffuse type) and involving the upper stomach (P < 0.05). EBNA1 and LMP2A expression were detected in 42 (93.3%) and 24 (53.3%) cases, respectively. None expressed EBNA2, LMP1, and ZEBRA. Among 45 cases of EBV associated gastric carcinomas, DNMT1, p16 and cyclin D1 expression were seen in 35 (77.8%), 10 (22.2%), and 29 (64.4%) cases, respectively. In contrast, among 40 EBV negative gastric carcinomas, expression of the three proteins were 20 (50.0%), 25 (62.5%) and 12 (30.0%), respectively. The difference of expression of the three proteins between the two groups was significant (P < 0.05). Expression of p16 correlated with the depth of the tumor invasion. Correlated protein expression was seen between LMP2A and DNMT1, between DNMT1 and p16, and between p16 and cyclin D1 (P < 0.05).</p><p><b>CONCLUSIONS</b>EBV associated gastric carcinoma accounts for 6.7% of gastric carcinomas in Guangzhou with the Latency I pattern in some cases and between Latency I and II in others. The correlated expression of LMP2A, DNMT1, p16 and cyclin D1 may contribute to the pathogenesis of EBV associated gastric carcinomas.</p>

Follicular dendritic cell sarcoma: a clinicopathologic analysis of ten cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features of follicular dendritic cell sarcoma (FDCS) and its differential diagnosis.</p><p><b>METHODS</b>Ten cases of FDCS were studied by light microscopy, immunohistochemistry and in-situ hybridization. The clinical features and follow-up information were analyzed.</p><p><b>RESULTS</b>Amongst the 10 cases of FDCS studied, the male-to-female ratio was 1:1. The mean age of the patients was 42 years. Six of them were located in cervical and peritoneal lymph nodes and four in extranodal sites (including tonsil, pelvic cavity, tail of pancreas and spleen). Histologically, the tumor cells had whorled, storiform or diffuse growth patterns. They were spindle in shape and contained syncytial eosinophilic cytoplasm, with round or oval nuclei, vesicular chromatin, distinct nucleoli and a variable number of mitotic figures. Multinucleated tumor giant cells and intranuclear pseudoinclusions were occasionally seen. There was a sprinkling of small lymphocytes and neutrophils within the tumor as well as in the perivascular region. Immunohistochemical study showed that the tumor cells were diffusely or focally positive for CD21, CD23, CD35 and D2-40, but negative for LCA, CD20, CD3, CD1a, HMB45 and CK. Some of them showed EMA, CD68 and S-100 reactivity. In-situ hybridization for Epstein-Barr virus-encoded RNA (EBER) showed positive signals in only one case (which was diagnosed as inflammatory pseudotumor-like FDCS). Of the 7 patients with follow-up information available (duration: 2 months to 39 months; mean: 14 months), 2 cases with paraneoplastic pemphigus died of pulmonary infection at 5 and 7 months respectively. The remaining 5 patients were alive and disease-free after surgical excision (+/- chemotherapy and radiotherapy).</p><p><b>CONCLUSIONS</b>FDCS is a rare low to intermediate-grade malignant tumor. Appropriate application of FDC markers, such as CD21, CD35 and D2-40, would be helpful for arriving at a correct diagnosis. Most cases are associated with good prognosis after surgical treatment, with or without chemotherapy and radiotherapy. Patients with paraneoplastic pemphigus carry a less favorable prognosis.</p>

Clinicopathologic study of 963 cases of mature T-cell and natural killer/T-cell lymphoma with respect to 2008 WHO classification of lymphoid neoplasms / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features of various types of mature T-cell and natural killer (NK)/T-cell lymphoma in Guangdong, China, with respect to the 2008 WHO classification of lymphoid neoplasms.</p><p><b>METHODS</b>Eleven hundred and thirty-seven (1137) cases of mature T-cell or NK/T-cell lymphoma diagnosed during the period from 2002 to 2006 in Guangzhou area were retrieved. The clinical data, histologic features and immunohistochemical findings were reviewed by a panel of experienced hematopathologists. Additional immunostaining was performed if indicated. The cases were re-classified according to the 2008 WHO classification of lymphoid neoplasms.</p><p><b>RESULTS</b>Nine hundred and sixty-three (963) cases fulfilled the diagnostic criteria of mature T-cell or NK/T-cell lymphoma and accounted for 20.1% of all cases of lymphoma encountered during the same period (963/4801). A predominance of extranodal involvement was noted in 644 cases (66.9%), while 319 cases (33.1%) showed mainly nodal disease. The prevalence of various lymphoma subtypes was as follows: peripheral T-cell lymphoma, unspecified (PTCL, NOS) 293 cases (30.4%), extranodal NK/T-cell lymphoma, nasal type 281 cases (29.2%), anaplastic large cell lymphoma (ALCL) 198 cases (20.6%), and angioimmunoblastic T-cell lymphoma (AILT) 46 cases (4.8%). The male-to-female ratio was 1.99. The median age of the patients was 44 years, with the peak age of PTCL, NOS, extranodal NK/T-cell lymphoma, nasal type and AILT being 55 to 64 years, 25 to 54 years and 65 to 74 years, respectively. ALK-positive ALCL occurred more frequently in young age, while the ALK-negative ALCL cases occurred mainly in the elderly.</p><p><b>CONCLUSIONS</b>Extranodal lesions predominate in mature T-cell and NK/T-cell lymphomas occurring in Guangzhou area. There is a male predominance and the overall incidence shows no increasing trend with age of the patient. The peak age of various subtypes however varies. The most common subtype was PTCL, NOS, followed by extranodal NK/T-cell lymphoma, nasal type, ALCL and AILT. The relatively frequent occurrence of extranodal NK/T-cell lymphoma, nasal type in Guangdong area is likely associated with the high incidence of Epstein-Barr virus infection there.</p>

Tumor necrosis factor-related apoptosis-inducing ligand induces glioma U87 cell apoptosis through the mitochondrial apoptotic pathway / 中华神经医学杂志

Objective To study the mechanism of tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)-induced apoptosis of glioma U87 cells.Methods Human glioma U87 cells were treated with human recombinant soluble TRAIL(rsTRAIL),and the cell apoptosis was detecmd with flow cytometry with AnnexinV-FITC/PI double staining.Flow cytometry with DiOC6 staining was used to assess the changes in mitochondrial transmembrane potential(△ψm).The relative activity of caspase-3,-8 and-9 Was measmed by colorimetric assay,and the concentration of cytoplasmic cytochrome C(cyt C) determined using enzyme-linked immunosorbem assay.The effects ofcaspase-8 inhibitor(Z-IETD-fmk)on rsTRAIL-induced apoptosis,△ψm,caspase-3,-8 and-9 activities and cyt C concentration were observed. Results RsTRAIL tinle-dependently induced apoptosis and progressive collapse of △ψm in glioma U87 cells,resulting also in caspase-3,-8 and-9activation and elevated cytC concentration.Caspase-8 inhibitor partially antagonized these biological effects induced by rsTRAIL in U87 cells.Conclusion TRAIL initiates a cascade of mitochondrial events by activating caspase-8 and induces apoptosis of glioma U87 cells.

Expressions of matrix metalloproteinase 9 in mucosal natural killer/T cell and mature T cell lymphomas and its relation to Epstein-Barr virus infection / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of matrix metalloproteinase 9 (MMP9) in mucosal natural killer/T cell and mature T cell lymphomas and its relation to Epstein-Barr virus (EBV) infection.</p><p><b>METHODS</b>The expression of MMP9 and EBV-encoded RNA (EBER) were detected by immunohistochemistry and in situ hybridization in 59 cases of mucosal natural killer/T cell and mature T cell lymphomas.</p><p><b>RESULTS</b>The positivity rates of MMP9 and EBERs were 83.05% and 72.88% respectively. The positivity rate of EBERs was correlated with histopathological subtype (P<0.05), but not with clinical stage, vascular invasion or the patients' survival time (P>0.05). The expression level of MMP9 was not correlated with the clinical stage, vascular invasion or survival time (P>0.05). No significant correlation was found between MMP9 expression and EBV infection.</p><p><b>CONCLUSION</b>EBV may play an important role in the development of mucosal natural killer/T cell and mature T cell lymphomas and promote disease progression by up-regulating MMP9 expression indirectly. Elimination of EBV infection may be helpful to prevent the development of lymphoma.</p>

A clinicopathological analysis of liver retransplantation in 22 cases / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To analyze the clinical and pathological features of patients with liver retransplantation.</p><p><b>METHODS</b>The clinical and pathological data of 22 patients who had liver retransplantation at our center from October 2003 to June 2006 were retrospectively analyzed.</p><p><b>RESULTS</b>Among the 523 patients who underwent liver transplantation, 22 (4.4%) had liver retransplantation. The causes of liver retransplantation were biliary tract complications (13/22), hepatic artery thrombosis (3/22), recurrence of hepatocellular carcinoma (5/22) and nonfunctional primary graft (1/22). The pathological changes in the livers of patients with biliary complications were intrahepatic cholestasis, primary bile duct hyperplasia and neutrophil infiltrations.</p><p><b>CONCLUSION</b>Biliary tract complications are the main cause of liver retransplantation. Differential diagnosis of various complications through early liver puncture biopsy and imaging examination will contribute to guide clinical treatment and may help in avoiding liver retransplantation.</p>

Expressions of h-TERT, c-myc, PCNA and cell apoptosis in liver carcinogenesis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expressions of human telomerase reverse transcriptase (h-TERT), c-myc, and proliferating cell nuclear antigen (PCNA) in chronic viral hepatitis (CVH), liver cirrhosis and primary hepatocellular carcinoma (HCC) and understand their possible role in liver carcinogenesis.</p><p><b>METHODS</b>Totally 157 liver disease specimens were collected, including 56 CVH, 52 liver cirrhosis and 49 primary HCC specimens. In situ hybridization was performed on these specimens to examine the expressions of h-TRET and c-myc mRNA, and immunohistochemistry carried out for PCNA detection, with the cell apoptosis detected with in situ ending labeling.</p><p><b>RESULTS</b>In the CVH, liver cirrhosis and primary HCC specimens, h-TERT expression was detected at the frequencies of 11/56 (19.6%), 43/52 (82.7%) and 44/47 (93.6%), c-myc expression at 7/56 (12.5%), 21/52 (40.4%) and 26/47 (55.3%), with apoptotic index of (27.3-/+4.7)%, (16.5-/+2.6)% and (8.7-/+1.3)% and PCNA expression rate of (17.1-/+2.9)%, (49.3-/+7.8)% and (62.5-/+9.1)%, respectively. Correlations among h-TERT, c-myc, and PCNA expressions and the apoptotic index were not found in the examined tissues (P>0.05).</p><p><b>CONCLUSION</b>Liver carcinogenesis may involve increased h-TERT, c-myc, and PCNA expressions and suppressed cell apoptosis.</p>

Relationship between clinicopathologic features and neoplasm recurrence,prognosis in hepatocellular carcinoma after liver transplantation / 中华器官移植杂志

Objective To study the risk factors affecting neoplasm recurrence and survival of hepatocellular carcinoma(HCC)following liver transplantation(LT).Methods The clinicopatholo gic data,neoplasm recurrence and survival results of 118 patients with HCC receiving LT were retro- spectively analyzed and various clinicopathologic risk factors for neoplasm recurrence and survival were evaluated by univariate and multivariate analysis.Results The follow-up time ranged from 1 to 32 months.The recurrence rate was 37.3% and the mortality was 35.5%.The 12-,18-,24-month survival rate was 84.55%,70.30% and 62.24%,respectively.The 12-,18-,24-month neoplasm free survival rate was 69.05%,66.93% and 61.38%,respectively.In the univariate analysis,por- tal vein neoplasm thrombus(PVTT),Milan-criteria,neoplasm size,histological differentiation and pTNM stage were associated significantly with neoplasm recurrence,and PVTT,Milan-criteria,pre- operative alpha-fetoprotein(AFP),histological differentiation and pTNM stage were associated signif- icantly with survival rate;In the multivariate Logistic regression analysis,PVTT and histological dif- ferentiation were independent predictive factors of neoplasm recurrence,and multivariate Cox regres- sion analysis showed that PVTT and AFP independently associated with prognosis.Conclusions PVTT and histological differentiation are the most important predictive factors of neoplasm recur- rence,and PVTT and AFP independently predict the survival of patients undergoing LT.