Pathological characteristics of liver allografts from donation after brain death followed by cardiac death in pigs / 华中科技大学学报（医学）（英德文版）

Donation after brain death followed by circulatory death (DBCD) is a unique practice in China. The aim of this study was to define the pathologic characteristics of DBCD liver allografts in a porcine model. Fifteen male pigs (25-30 kg) were allocated randomly into donation after brain death (DBD), donation after circulatory death (DCD) and DBCD groups. Brain death was induced by augmenting intracranial pressure. Circulatory death was induced by withdrawal of life support in DBCD group and by venous injection of 40 mL 10% potassium chloride in DCD group. The donor livers were perfused in situ and kept in cold storage for 4 h. Liver tissue and common bile duct samples were collected for hematoxylin and eosin staining, TUNEL testing and electron microscopic examination. Spot necrosis was found in hepatic parenchyma of DBD and DBCD groups, while a large area of necrosis was shown in DCD group. The apoptosis rate of hepatocytes in DBD [(0.56±0.30)%] and DBCD [(0.50 ± 0.11)%] groups was much lower than that in DCD group [(3.78±0.33)%] (P0.05)). The structures of bile duct were intact in both DBD and DBCD groups, while the biliary epithelium was totally damaged in DCD group. Under electron microscope, the DBD hepatocytes were characterized by intact cell membrane, well-organized endoplasmic reticulum, mild mitochondria edema and abundant glycogens. Broken cell membrane, mild inflammatory cell infiltration and sinusoidal epithelium edema, as well as reduced glycogen volume, were found in the DBCD hepatocytes. The DCD hepatocytes had more profound cell organelle injury and much less glycogen storage. In conclusion, the preservation injury of DBCD liver allografts is much less severe than that of un-controlled DCD, but more severe than that of DBD liver allografts under electron microscope, which might reflect post-transplant liver function to some extent.

Pathological characteristics of liver allografts from donation after brain death followed by cardiac death in pigs / 华中科技大学学报（医学）（英德文版）

Donation after brain death followed by circulatory death (DBCD) is a unique practice in China. The aim of this study was to define the pathologic characteristics of DBCD liver allografts in a porcine model. Fifteen male pigs (25-30 kg) were allocated randomly into donation after brain death (DBD), donation after circulatory death (DCD) and DBCD groups. Brain death was induced by augmenting intracranial pressure. Circulatory death was induced by withdrawal of life support in DBCD group and by venous injection of 40 mL 10% potassium chloride in DCD group. The donor livers were perfused in situ and kept in cold storage for 4 h. Liver tissue and common bile duct samples were collected for hematoxylin and eosin staining, TUNEL testing and electron microscopic examination. Spot necrosis was found in hepatic parenchyma of DBD and DBCD groups, while a large area of necrosis was shown in DCD group. The apoptosis rate of hepatocytes in DBD [(0.56±0.30)%] and DBCD [(0.50 ± 0.11)%] groups was much lower than that in DCD group [(3.78±0.33)%] (P<0.05). And there was no significant difference between DBD group and DBCD group (P>0.05)). The structures of bile duct were intact in both DBD and DBCD groups, while the biliary epithelium was totally damaged in DCD group. Under electron microscope, the DBD hepatocytes were characterized by intact cell membrane, well-organized endoplasmic reticulum, mild mitochondria edema and abundant glycogens. Broken cell membrane, mild inflammatory cell infiltration and sinusoidal epithelium edema, as well as reduced glycogen volume, were found in the DBCD hepatocytes. The DCD hepatocytes had more profound cell organelle injury and much less glycogen storage. In conclusion, the preservation injury of DBCD liver allografts is much less severe than that of un-controlled DCD, but more severe than that of DBD liver allografts under electron microscope, which might reflect post-transplant liver function to some extent.

Serum proteomic variation study in patients with Crohn disease / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To search differentially expressed proteins in serum of patients with Crohn disease.</p><p><b>METHODS</b>Serum protein samples from 4 patients with Crohn disease and 8 healthy adults were recruited cross-labeled with variant CyDye, and then followed by two-dimensional differential in-gel electrophoresis (2-D DIGE), image analysis, and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).</p><p><b>RESULTS</b>The 2-D electrophoresis results were compared between the Crohn disease patients and the healthy adults. The spot 1058 expression in serum of Crohn disease patients increased by 1.68 folds as compared with healthy adults (P<0.05). The protein was identified as haptoglobin by mass spectrometry.</p><p><b>CONCLUSION</b>Up-regulating expression of haptoglobin in serum of Crohn disease patients may play a role in disequilibrium of immunity system.</p>

Expression of survivin mRNA and protein in mitomycin-treated hepatoma carcinoma Hepa1-6 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate survivin mRNA and protein expressions in mitomycin (MMC)-treated hepatoma carcinoma Hepa1-6 cells in vitro.</p><p><b>METHODS</b>Hepa1-6 cells were cultured in vitro in the presence of MMC at the concentrations of 1.0, 3.0 and 9.0 microg/ml, respectively, and 1 day and 3 days after the culture, the cell growth inhibition was assessed using MTT assay and the expressions of survivin were detected by RT-PCR and Western blotting.</p><p><b>RESULTS</b>MMC at the concentration of 9.0 microg/ml resulted in significantly greater growth inhibition of the Hepa-6 cells than MMC at 1.0 and 3.0 microg/ml, and at the latter two concentrations, MMC treatment for 3 days did not produce obvious cell growth inhibition. Survivin expressions at both the mRNA and protein levels in Hepa1-6 cells were significantly decreased 1 day after MMC treatment at the 3 concentrations, and after 3-day MMC treatment at 1.0 and 3 microg/ml, survivin expressions increased to exceed the control level, whereas survivin maintained the low expression levels in cells treated with 9 microg/ml MMC for 3 days.</p><p><b>CONCLUSION</b>Survivin expression in Hepa1-6 cells increases in response to MMC treatment at low doses, which might be one of the reasons for chemotherapeutic drug resistance.</p>

Increased rate change over time of a sphincter-saving procedure for lower rectal cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Total mesorectal excision (TME) has increased the rate of sphincter-preservation (SP) for more patients with low-lying rectal cancer. Here, we analyze the change of sphincter preserving rates in lower rectal cancer and their related factors.</p><p><b>METHODS</b>We reviewed retrospectively the medical records of 316 patients with lower rectal cancers, 1 to 5 cm from the anorectal line, who had surgical resections from August 1994 to November 2005. The 12-year span was divided into 2 periods: period I (August 1994-December 1998) and period II (January 1999-November 2005), based on the date (January 1999) when standard total mesorectal excision (TME) was introduced. The patients were divided into two groups based on the operation: abdominoperineal resection (APR) or SP surgery. SP rates, leakage and other clinico-pathological characteristics were compared between the two time periods and between the two different groups.</p><p><b>RESULTS</b>The SP rate increased significantly over the 12 years, from 44.9% in period I to 76.2% in period II (P = 0.000). The factors significantly influencing SP included the distance of the tumor from the anorectal line, gender, time period, circumference of intramural spread and histological differentiation (P < 0.05). Significant differences were detected between the two time periods in gender, blood transfusion volume and Dukes' stage (P < 0.05). The leakage rate was 2.7% in period I and 1.3% in period II (P > 0.05).</p><p><b>CONCLUSIONS</b>Over the 12-year period of the study the SP rate in rectal cancers 1 - 5 cm from the anorectal line has increased significantly while the blood transfusion volume has decreased due to the introduction of TME. However, TME had no effect on operating time and leakage rates.</p>

Correlations of expressions of macrophage migration inhibitor factor and cyclin D1 with tumor size and metastasis of hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore the possible relationship between the expressions of macrophage migration inhibitor factor (MIF), cyclin D1, cyclin-dependent kinase 4 (CDK4), phosphorylated-retinoblastoma susceptibility gene product Rb protein (phospho-Rb) and the development of hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>93 HCC tissues and 5 normal liver tissues were used to investigate the expressions of MIF, cyclin D1, CDK4 and phospho-Rb by tissue microarray and immunohistochemistry methods.</p><p><b>RESULTS</b>The expression rates of MIF, cyclin D1, CDK4 and phospho-Rb in the HCC tissues were 71%, 41%, 82% and 14% respectively, and in the normal liver tissues, they were 0%, 0%, 80% and 20% respectively. The expression rates of MIF and cyclin D1 were significantly different between the tumor and the normal liver tissues and the expression rates of CDK4 and phospho-Rb were not significantly different between the tumor and the normal liver tissues. The rate difference (69% versus 48%) of MIF expression between the larger tumors (> 3.5 cm) and the smaller tumors (< 3.5 cm) was of statistical significance (P < 0.01). The expression rate (62%) of cyclin D1 in the tumors with metastasis was significantly higher than the expression rate (35%) in the tumors without metastasis (P < 0.05). MIF expression was positively correlated with cyclin D1 expression in the tumor tissues (P < 0.01). CDK4 and phospho-Rb expressions were not significantly associated with the tumor sizes and metastasis status.</p><p><b>CONCLUSION</b>Our results indicate that MIF and cyclin D1 might be related to the growth and metastasis of HCC.</p>

Construction and identification of human bone morphogenetic protein-7 recombinant adeno-associated virus type 2 vector and its expression in bone mesenchymal stem cells / 中华外科杂志

<p><b>OBJECTIVES</b>To facilitate gene therapy research using recombinant adeno-associated virus type 2 (rAAV2) vector as gene transfer vehicle, and to construct a rAAV2 based vector carrying bone morphogenetic protein-7 (BMP7) and observe its expression in bone mesenchymal stem cells.</p><p><b>METHODS</b>The coding sequence (1.3 kb) of BMP7 was amplified by polymerase chain reaction (PCR) from the pcDNA1.1(+) plasmid containing the human BMP-7 cDNA. After purified, the gene fragment was cloned into a plasmid pUC18 and termed plasmid pUC18-hBMP7. The recombinant pUC18-hBMP7 was digested by Kpn I and Sal I and further ligated to the pSNAV by T4DNA ligase. The resultant plasmid PSNAV-hBMP7 was transformed into DH5a Escherichia coli, and positive colonies were screened by PCR and digest with restriction enzyme to identify the correct recombinant clones. BHK-21 cells were transfected with the purified pSNAV-BMP7 plasmid according to a standard calcium phosphate precipitation method. The cells were then cultured in selection media containing 800 micro g/ml G418 (Gibco/BRL). G418-resistant BHK-21 cell clones were isolated and the integrity of hBMP7 gene was determined by PCR using the above PCR primers. To package the virus, stably transfected BHK-21 cells were subsequently infected with recombinant herpes simplex virus type 1 (rHSV-1). The collected cells were processed by chloroform treatment, PEG8000/NaCl precipitation and chloroform extraction for purification. The titer was determined using quantitative DNA dot blots and the purity was examined by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Following infection with rAAV2-BMP7 at multiplicities of infection of 1 x 10(5) vector genomes per cell and subsequent culture, MSCs were assessed qualitatively for BMP7 production.</p><p><b>RESULTS</b>Transient transfection showed an efficiency of 98.8% in MSCs. RT-PCR showed that MSCs had transcription of BMP7 that was enhanced by the gene transfer. BMP-7 expression in MSCs was identified by Western-blot.</p><p><b>CONCLUSIONS</b>The hBMP7 recombinant adeno-associated virus vector is successfully constructed. The present in vitro study demonstrates that rAAV2-BMP7 could infect MSCs.</p>

Arginine supplementation in total parenteral nutrition improves postoperative cellular immune function of patients with hepatocellular carcinoma after operation / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the effect of total parenteral nutrition (TPN) supplemented with arginine on cellular immune function of patients with hepatocellular carcinoma (HCC) after radical tumor resection.</p><p><b>METHODS</b>Fifty-six HCC patients undergoing radical surgery received fat-free TPN support, routine TPN or TPN with arginine supplementation, and their clinical data were analyzed prospectively. The percentages of T lymphocyte subpopulation and national killer (NK) cells in peripheral blood are determined, and the levels of interleukin-2 (IL-2), IL-4 and interferon-gamma (IFN-gamma) were measured.</p><p><b>RESULTS</b>No marked changes were noted in peripheral blood CD4+, CD8+ T cells and NK cells, or in IL-2, IL-4 and IFN-gamma levels after fat-free TPN and routine TPN support. TPN supplemented with arginine resulted in significant increase in CD4+ T cells, NK cells and CD4+/CD8+ T cell ratio in the peripheral blood, as well as in IL-2 and IFN-gamma levels. Peripheral blood IL-4 level was decreased significantly.</p><p><b>CONCLUSION</b>TPN with arginine supplementation can augment the percentages of CD4+ T lymphocytes and NK cells, and increase IL-2 and IFN-gamma levels, suggesting that arginine can enhance cell-mediated immunity in postoperative patients with HCC.</p>

Experimental study on effect of dexamethasone to the in-stent restenosis after vascular intervention / 介入放射学杂志

Objective To evaluate the effect of dexamethasone to the cultured rat thoracic aortic smooth muscle cells(SMC)in vitro,and explore the role on it's prevention and cure for the in-stent restenosis after vascular intervention.Methods The rat thoracic aortic SMC were harvested and cultured for six to ten passages.The cultured SMC were synchronized and then restimulated to enter the cell cycle,and treated with incremental concentrations of dexamethasone or without dexamethasone as control.The proliferative assay was performed with MTT method in the different time points after treatment.RT-PCR was performed to assay the level of proliferating cell nuclear antigen(PCNA)mRNA.Results 1.Dexamethasone progressively inhibited rat aortic SMC proliferation in a concentration-dependent fashion.The A value was statistically significant for different concentrations(F=36.02,P＜0.001).The effect was not significant for dexamethasone concentrations either between 10~(-6)and 10~(-5)mol/L(P=0.065)or between 10~(-11)mol/L and control group(P= 0.567).2.RT-PCR suggested dexamethasone significantly decreased rat aortic SMC PCNA mRNA transcription in a concentration-dependent fashion.Statistical analysis indicated F=15.407 and P＜0.001 by ANOVA. Comparing to the control,the corrected A value was not statistically significant at 10~(-9)or 10~(-11)mol/L groups by post hoc analysis.Conclusions Dexamethasone inhibits rat aortic SMC proliferation in a concentration- dependent fashion.The data suggest that effective action concentration is 10~(-7)mol/L with persistent time up to 96 hours or more.Dexamethasone may play the inhibit role to SMC at lower concentration with prolonging action time.