Study of Recellularized Human Acellular Arterial Matrix Repairs Porcine Biliary Segmental Defects

BACKGROUND: With the popularity of laparoscopic cholecystectomy, common bile duct injury has been reported more frequently. There is no perfect method for repairing porcine biliary segmental defects.METHODS: After the decellularization of human arterial blood vessels, the cells were cultured with GFP⁺ (carry green fluorescent protein) porcine bile duct epithelial cells. The growth and proliferation of porcine bile duct epithelial cells on the human acellular arterial matrix (HAAM) were observed by hematoxylin-eosin (HE) staining, electron microscopy, and immunofluorescence. Then, the recellularized human acellular arterial matrix (RHAAM) was used to repair biliary segmental defects in the pig. The feasibility of it was detected by magnetic resonance cholangiopancreatography, liver function and blood routine changes, HE staining, immunofluorescence, real-time quantitative PCR (RT-qPCR), and western blot.RESULTS: After 4 weeks (w) of co-culture of HAAM and GFP? porcine bile duct epithelial cells, GFP⁺ porcine bile duct epithelial cells grew stably, proliferated, and fused on HAAM. Bile was successfully drained into the duodenum without bile leakage or biliary obstruction. Immunofluorescence detection showed that GFP-positive bile duct cells could still be detected after GFP-containing bile duct cells were implanted into the acellular arterial matrix for 8 w. The implanted bile duct cells can successfully resist bile invasion and protect the acellular arterial matrix until the newborn bile duct is formed.CONCLUSION: The RHAAM can be used to repair biliary segmental defects in pigs, which provides a new idea for the clinical treatment of common bile duct injury.

An enzyme-linked immunosorbent assay for determining serum anti-themocyte globulin concentration / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish an enzyme-linked immunosorbent assay (ELISA) for determining anti-themocyte globulin (ATG) levels in serum samples.</p><p><b>METHODS</b>The microplate was coated with mouse anti-rabbit IgG monoclonal antibody, and sheep anti-rabbit polyclonal antibody conjugated with HRP was used as the second antibody for detecting the serum ATG levels in patients undergoing allogeneic hematopoietic stem cell transplantation.</p><p><b>RESULTS</b>The optimal concentration of the coating antibody and dilution ratios of the serum samples and IgG-HRP conjugate were 0.2 microg/ml, 1:40 and 1:2500, respectively. The lower sensitivity limit of the assay was 31.25 ng/ml for ATG detection. A linear relationship was established within the concentration range from 40 to 1000 ng/ml, with the coefficients of variation of 7.91 within assay and 5.22 between assays, respectively. Seven patients undergoing stem cell transplantation with ATG pretreatment showed gradually decreased concentration of ATG, and after 90 days ATG could still be detected.</p><p><b>CONCLUSION</b>The sandwich ELISA we established provides a specific and sensitive method for quantitative measurement of ATG in the clinical setting. In patients undergoing stem cell transplantation with ATG pretreatment, the ATG concentration gradually decreases but remains detectable 90 days after the administration.</p>

Value of combining 64 multi-slice spiral computer tomography and serum amyloid A protein in surgical decision-making in rectal cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To determine the accuracy and clinical value of combining 64 multi-slice spiral computer tomography (MSCT) and serum amyloid A protein (SAA) in the preoperative staging of rectal cancer.</p><p><b>METHODS</b>Prospectively enrolled patients with rectal cancer from October 2007 to October 2008. The patients were randomly assigned into two groups: MSCT and SAA combined group: both MSCT and SAA combinative assessment were performed for preoperative evaluation; MSCT group: only MSCT was performed preoperatively for tumor staging. The accuracy of the preoperative T, N, M, and TNM staging and the concordance rate of predictive operative strategy were compared between the two groups.</p><p><b>RESULTS</b>Total of 225 cases with rectal cancer were enrolled in this study. There were 110 cases in MSCT and SAA combined group and 115 cases in MSCT group. The baseline characteristics was comparable between the two groups. For MSCT and SAA combined group, the accuracies of preoperative staging of T, N, M and TNM was 87.3%, 85.2%, 100% and 86.4%, respectively; and for MSCT group, the corresponding rates was 85.2%, 67.0%, 100% and 66.1%, respectively. Statistical differences was found in the accuracy of preoperative N and TNM staging between the two groups (P = 0.009 and 0.001, respectively). In addition, there was statistical difference in the accuracy of prediction to operative procedures between the two groups (94.7% vs. 81.7%, P = 0.003).</p><p><b>CONCLUSION</b>Combinative assessment of MSCT and SAA could improve the accuracy of preoperative staging, and thus provide higher predictive coincidence rate of operative procedures.</p>