Protective effects of affiliating portasystemic shunt on graft injury in small-for-size liver transplantation in miniature pigs / 中华外科杂志

<p><b>OBJECTIVES</b>To evaluate the protective effects of affiliating portasystemic shunt on small-for-size graft in liver transplantation.</p><p><b>METHODS</b>Fifteen Chinese Bama miniature pigs were divided into three groups: group A (small-for-size liver transplantation), group B (distal splenorenal shunt + small-for-size liver transplantation), and group C (mesocaval H-shape shunt + small-for-size liver transplantation). Animals were followed up for 7 days with survival, dynamical liver function biochemical parameters, liver biopsies, portal venous pressure (PVP) and portal blood flow (PBF).</p><p><b>RESULTS</b>Animal survivals were as follows: group A, 1/5, group B, 3/5 and group C, 5/5.Group A resulted in abnormal liver function parameters that were significantly ameliorated in group B and C. The histological examination of graft in group A displayed severe pathologic changes including hepatocyte vacuolar change or necrosis, sinusoidal congestion, parenchymal hemorrhage. Affiliating portasystemic shunt significantly alleviated graft injuries in group B and C. PVP rose and peaked up to 28.6 mm Hg (1 mm Hg = 0.133 kPa), PBF fluctuated after reperfusion in group A, but group B and C with affiliating portasystemic shunt showed significantly lower PVP and maintained rather stable PBF after reperfusion. There were also statistical differences in PVP or PBF between group B and C.</p><p><b>CONCLUSIONS</b>Affiliating portasystemic shunt effectively might protect small-for-size graft from injuries after reperfusion.</p>

Surgical treatment for hilar cholangiocarcinoma of Bismuth-Corlette type IV / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the surgical treatment for hilar cholangiocarcinoma(HCCA) of Bismuth-Corlette type IV.</p><p><b>METHODS</b>The clinical data of 73 HCCA patients admitted in Southwest Hospital, the Third Military Medical University from January 2002 to December 2008 were analyzed retrospectively. There were 41 males and 32 females, aged from 30 to 84 years old (averaged, 56.8 years old). All patients were diagnosed as hilar mass with hilar biliary obstruction by B-ultrasound, CT, magnetic resonance imaging, endoscopic retrograde cholangiopancreatography, magnetic resonance cholangiopancreatography or percutaneous transhepatic cholangiography, and confirmed by pathological examination in intra-operation and post-operation. Diagnosis of all patients were according to Bismuth-Corlette type IV.</p><p><b>RESULTS</b>Fifteen cases underwent percutaneous transhepatic cholangial drainage, stents were implanted in 8 patients. Simple internal drainage was performed on 25 of the 73 patients and 4 with palliative resection. Radical resection was performed on 19 patients. The radical resection rate of HCCA were 26.0%. The 1, 3 years of survival rates were 36.8%, 10.5% respectively. The 1 year survival rate was 6.3% in drainage group.</p><p><b>CONCLUSION</b>Radical resection is the potentially curative treatment for HCCA, a sufficient, reasonable use of internal and external drainage would improve the patient's quality of life.</p>

Immune response of dendritic cells acquiring antigens from apoptotic cholangiocarcinoma cells induced by mitomycin / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the anti-tumor immune response of dendritic cells (DCs) acquiring antigens from apoptotic cholangiocarcinoma cells and their therapeutic effects on cholangiocarcinoma cells.</p><p><b>METHODS</b>DCs from human peripheral blood monocytes which acquired antigen capturing and processing capacity, characteristics of maturation, were established in vitro using granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). Then cholangiocarcinoma cells were induced to apoptosis with mitomycin. The three groups included (1) coculture of DCs, apoptotic cancer cells and T cells, (2) coculture of DCs, necrotic cancer cells and T cells, (3) coculture of DCs, cultured cancer cells and T cells. After 7 days, DCs and T cells were riched separately to perform anti-tumor cells test and immune response test.</p><p><b>RESULTS</b>these cells had typical dendritic cell morphology, expressed high levels of CD1a and B7, acquired antigen from apoptotic cells caused by mitomycin and could stimulate T cells to inhibit, even kill cholangiocarcinoma cells.</p><p><b>CONCLUSIONS</b>The DCs from peripheral blood monocytes induced by GM-CSF and IL-4 can efficiently present antigen derived from apoptotic cells caused by mitomycin, and stimulate T cells activity obviously. It maybe become an effective therapy for tumor.</p>

Screening and identification for cDNA of differentially expressed genes in human primary hepatocellular carcinoma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>Screening and identification of differentially expressed genes in human primary hepatocellular carcinoma(HCC).</p><p><b>METHODS</b>The differentially expressed genes subtracted cDNA library of HCC constructed by suppression subtractive hybridization(SSH) technique was screened by colony in situ hybridization, then the positive clones were further screened with PCR amplification. The positive clones were sequenced and analyzed for homology in the Genbank databases with Basic Local Alignment Search Tool BLAST . The novel cDNA sequences were analyzed by Northern blot analysis.</p><p><b>RESULTS</b>Thirteen positive clones were obtained, and 11 cDNA sequences were identified. Sequences of 11 cDNA showed that 6 cDNA were homologous with the genes published in Genbank and 5 cDNA were unknown genes. Northern blot indicated that 3 novel cDNA(>300 bp) were only expressed in HCC.</p><p><b>CONCLUSION</b>The subtracted cDNA library constructed by SSH technique contains differentially expressed genes of HCC. Three novel cDNA sequences might be differentially expressed genes of HCC. Further screening the library and gaining the whole gene sequence may lay a foundation for identifying differentially expressed genes in HCC.</p>

Clinical significances of growth hormone and growth hormone binding pro tein determination in patients with obstructive jaundice / 第三军医大学学报

Objective　To evaluate the changes and clinical s ignificances of serum growth hormore (GH) and GH binding protein (GHBP) in patie nts with obstructive jaundice (OJ). Methods　Serum levels of GH, GHBP, insulin-like growth factor-Ⅰ (IGF-Ⅰ), endotoxin, prealbumin concentr ation and liver function in 60 male in patients with OJ were detected 1 d preope ratively and 1, 3, 7 d postoperatively. And 40 male patients with simple gallbla dder stone hospitalized in the same period were served as control. Resu lts　The levels of GH, GHBP and IGF-Ⅰ were significantly lower in pati ents with obstructive jaundice than in control group(P<0.01), which was corr elated with the levels of bilirubin, endotoxin and prealbumin (P<0.01). Conclusion　The obviously downregulation of GH/GHBP axis in patients with OJ probably results from the inbibition of expression of GH receptor due t o nutritional dysfunction and endotoxemia, and further aggravate the nutritional dysfuction and endoxemia, which may be one of the important factors in the path ophysiological changes of OJ.

Expression of CD14 in Kupffer′s cells induced by Lipopolysaccharide / 第三军医大学学报

Objective　To investigate the expression of LPS receptor-CD14(CD14) on the membrane of Kupffer′s cells (KCs) induced by Lipopolysaccharide (LPS )and its role in activation of KCs and production of cytokines. Methods　KCs were isolated by collagenase perfused Wistar rats and routinely cultured in 24-well dishes for 12 h. Cells were harvested and adjusted to a concentration of 1×106/ml/well and were devided into two groups. Group of LPS: KCs were induced with different concentration of LPS (0, 100 ng/ml, 1 μg/ml and 100 μg/ml). Group of PI-PLC: KCs were pre-incubated for 30 min with one unit of phophatidy linositol specific phospholipase C (PI-PLC) before different concentrations of LPS were added. KCs were cultured for 30 and 60 min respectively. Supernatants were then collected for measuring the level of TNFα and IL-6. Cells were stained by indirect immunofluorescent method ( rabbit anti-CD14 antibody and goat anti-rabbit IgG conjugated with FITC ) and analyzed with flow cytometer (FCM). The percentage and mean fluorescence intensity (FI) of CD14-positive cells were taken as the indexes. Results　In LPS group, after incubation of cells with increasing concentration of LPS, a significant increase in the percentage of CD14 positive KCs were found and the mean FI was stronger when compared with the control points or the group of PI-PLC. The levels of TNFα and IL-6 in supernatant also increased (P<0.01). In group of PI-PLC, decreased percentage of CD14 positive KCs and weakened mean FI were found when compared with group of LPS. The increasing production of TNFα and IL-6 slowed down in the group of PI-PLC. Conclusion　CD14 expression of KCs might be up-regulated by LPS with increase of some cytokines. The production of cytokines in KCs induced by LPS is partially inhibited by PI-PLC.

Expression of HGF/Met mRNA and TGF-α/EGFR mRNA in the liver/hepatocyte after partial hepatectomy in noncirrhotic obstructive rats / 第三军医大学学报

Objective　To investigate the expression of HGF and TGF-α and their receptor, Met (HGF receptor) and EGFR (TGF-αreceptor) mRNA, in the regenerative liver/hepatocytes after 70% partial hepatectomy (70% PH) in noncirrhotic biliary obstruction rats. Methods　Wistar rats were divided randomly into N-PH group, BDO-RBF-PH group and BDO-RBF group. The expression of HGF/Met mRNA and TGF-α/EGFR mRNA was measured by RT-PCR in the liver/hepatocytes at the time point of 0, 6, 12, 24, 48 and 72 h after 70% PH or RBF. Results　In N-PH group, the expression of HGF/Met mRNA increased sharply and peaked at 6 h, and maintained at a high level until 24 h after 70% PH. In BDO-RBF-PH group however, the expression of HGF/Met mRNA increased slowly and peaked at 12 h after 70% PH. The peak level was lower in BDO-RBF-PH group than in N-PH one. The expression of TGF-α/EGFR mRNA increased sharply and peaked at 24 h after 70% PH in N-PH group. However, the expression of TGF-α/EGFR mRNA elevated slowly and peaked at 48 h after 70% PH in BDO-RBF-PH group with a lower peak level than that in N-PH group. Conclusion　The expression of HGF/Met mRNA and TGF-α/EGFR mRNA in the regenerative liver/hepatocytes after 70% PH decreases significantly in noncirrhotic biliary duct obstruction rats. There is a tendency that the expression of HGF mRNA and TGF-α mRNA is less than Met mRNA and EGFR mRNA.

Clinical significances of growth hormone and growth hormone binding pro tein determination in patients with obstructive jaundice / 第三军医大学学报

Objective　To evaluate the changes and clinical s ignificances of serum growth hormore (GH) and GH binding protein (GHBP) in patie nts with obstructive jaundice (OJ). Methods　Serum levels of GH, GHBP, insulin-like growth factor-Ⅰ (IGF-Ⅰ), endotoxin, prealbumin concentr ation and liver function in 60 male in patients with OJ were detected 1 d preope ratively and 1, 3, 7 d postoperatively. And 40 male patients with simple gallbla dder stone hospitalized in the same period were served as control. Resu lts　The levels of GH, GHBP and IGF-Ⅰ were significantly lower in pati ents with obstructive jaundice than in control group(P<0.01), which was corr elated with the levels of bilirubin, endotoxin and prealbumin (P<0.01). Conclusion　The obviously downregulation of GH/GHBP axis in patients with OJ probably results from the inbibition of expression of GH receptor due t o nutritional dysfunction and endotoxemia, and further aggravate the nutritional dysfuction and endoxemia, which may be one of the important factors in the path ophysiological changes of OJ.

Expression of CD14 in Kupffer′s cells induced by Lipopolysaccharide / 第三军医大学学报

Objective　To investigate the expression of LPS receptor-CD14(CD14) on the membrane of Kupffer′s cells (KCs) induced by Lipopolysaccharide (LPS )and its role in activation of KCs and production of cytokines. Methods　KCs were isolated by collagenase perfused Wistar rats and routinely cultured in 24-well dishes for 12 h. Cells were harvested and adjusted to a concentration of 1×106/ml/well and were devided into two groups. Group of LPS: KCs were induced with different concentration of LPS (0, 100 ng/ml, 1 μg/ml and 100 μg/ml). Group of PI-PLC: KCs were pre-incubated for 30 min with one unit of phophatidy linositol specific phospholipase C (PI-PLC) before different concentrations of LPS were added. KCs were cultured for 30 and 60 min respectively. Supernatants were then collected for measuring the level of TNFα and IL-6. Cells were stained by indirect immunofluorescent method ( rabbit anti-CD14 antibody and goat anti-rabbit IgG conjugated with FITC ) and analyzed with flow cytometer (FCM). The percentage and mean fluorescence intensity (FI) of CD14-positive cells were taken as the indexes. Results　In LPS group, after incubation of cells with increasing concentration of LPS, a significant increase in the percentage of CD14 positive KCs were found and the mean FI was stronger when compared with the control points or the group of PI-PLC. The levels of TNFα and IL-6 in supernatant also increased (P<0.01). In group of PI-PLC, decreased percentage of CD14 positive KCs and weakened mean FI were found when compared with group of LPS. The increasing production of TNFα and IL-6 slowed down in the group of PI-PLC. Conclusion　CD14 expression of KCs might be up-regulated by LPS with increase of some cytokines. The production of cytokines in KCs induced by LPS is partially inhibited by PI-PLC.

Expression of HGF/Met mRNA and TGF-α/EGFR mRNA in the liver/hepatocyte after partial hepatectomy in noncirrhotic obstructive rats / 第三军医大学学报

Objective　To investigate the expression of HGF and TGF-α and their receptor, Met (HGF receptor) and EGFR (TGF-αreceptor) mRNA, in the regenerative liver/hepatocytes after 70% partial hepatectomy (70% PH) in noncirrhotic biliary obstruction rats. Methods　Wistar rats were divided randomly into N-PH group, BDO-RBF-PH group and BDO-RBF group. The expression of HGF/Met mRNA and TGF-α/EGFR mRNA was measured by RT-PCR in the liver/hepatocytes at the time point of 0, 6, 12, 24, 48 and 72 h after 70% PH or RBF. Results　In N-PH group, the expression of HGF/Met mRNA increased sharply and peaked at 6 h, and maintained at a high level until 24 h after 70% PH. In BDO-RBF-PH group however, the expression of HGF/Met mRNA increased slowly and peaked at 12 h after 70% PH. The peak level was lower in BDO-RBF-PH group than in N-PH one. The expression of TGF-α/EGFR mRNA increased sharply and peaked at 24 h after 70% PH in N-PH group. However, the expression of TGF-α/EGFR mRNA elevated slowly and peaked at 48 h after 70% PH in BDO-RBF-PH group with a lower peak level than that in N-PH group. Conclusion　The expression of HGF/Met mRNA and TGF-α/EGFR mRNA in the regenerative liver/hepatocytes after 70% PH decreases significantly in noncirrhotic biliary duct obstruction rats. There is a tendency that the expression of HGF mRNA and TGF-α mRNA is less than Met mRNA and EGFR mRNA.