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Objective To establish an ELISA for quantitative determination of decoy receptor 3 (DcR3) in human plasma.Methods A solid phase double antibody sandwich method was established for quantitative determination of DcR3.The anti-DcR3 antibody was immobilized onto ELISA plate.DcR3 in samples was captured by anti-DcR3 on ELISA plate and then detected by biotin-anti-DcR3 and subsequent peroxidase-labeled streptavidin,and the color was developed by adding substrate.The standard DcR3 samples on the same plate were detected simultaneously to calculate the DcR3 concentrations in unknown samples.The sensitivity,specificity,precision,recovery,linearity and DcR3 range in normal human adults were assessed.Results The sensitivity of the developed assay was 0.051 ng/mL.The intra-coefficient of variation (CV) was less than 10％ and inter-CV was less than 15％.The average recovery rate was 90.50％.When 2-fold amount of anti-TNF-α was added into the coated antibodies,10-fold amount of biotin-labeled anti-LIGHT,antiFAS or anti-TNF-α was added into the detection antibodies,or 10 fold amount of purified LIGHT protein was added into the standard DcR3 samples as competitor,no disturbing effects on standard curve were found.The linear range of the assay was from 0.25 to 16 ng/mL (r≥0.98).The concentration of DcR3 tested in 128 plasma samples from healthy adults was (0.21 ± 0.05) ng/mL with 95％ CI ranged from 0.14 to 0.28 ng/mL and no difference of age and sex was found.Conclusion The established ELiSA for determining plasma DcR3 exhibited high specificity,sensitivity,precision,fine linearity and wide detecting range.This method could be used for quantification of DcR3 in plasma.
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Objective To evaluate the therapeutic methods and the key points in the management of delayed complications of bile duct stenosis after iatrogenic bile duct injuries.Methods Five patients with bile duct injuries developed delayed complications of bile duct stenosis after cholecystectomy were retrospectively studied.All clinical information were derived from the medical data of these 5 patients treated in our department from June 2002 to July 2006.Results Of the 5 patients with delayed complications of iatrogenic bile duct injuries,2 patients developed common bile duct occlusion,2 patients developed hilar bile duct stenosis,and 1 patient developed anastomotic stenosis after cholangiojejunostomy.Bile duct injuries occurred in 3 patients after laparoscopic cholecystectomy,and in another 2 patients after mini-laparotomy-cholecystectomy.The locations of the bile duct injuries,according to Strasberg classification,were type E1 (n=1),type E2 (n=1),type E3,(n=1),and type E5 (n=2).All 5 patients received PTCD initially to relieve obstructive jaundice and then they received surgical exploration and biliary reconstruction.These patients received cholangioplasty at the porta hepatis,and hepaticojejunostomy using a Roux-en-Y anastomosis.Two patients received Roux-en-Y anastomosis of the common hepatic duct to the jejunum.All these patients had good recovery and were discharged well postoperatively.All patients had been followed-up for 7 years or more,with no evidence of obstructive jaundice or biliary tract infection.Conclusions For patients who present with delayed complications of bile duct stenosis after iatrogenic bile duct injuries,primary PTCD is carried out to relieve jaundice and to control infection.Meticulous dissection of the porta hepatis,reliable anastomosis of the proximal healthy bile duct tissues to a jejunal loop,and adequate size of anastomosis guarantee success of surgery.
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Objective To observe the effect on Foxg1 gene expression in the subgranular zone (SGZ) of cerebral tissue from neonatal rats with hypoxic-ischemic brain damage (HIBD) after transplantation of neural stem cells (NSCs) derived from umbilical cord blood.Methods Mononuclear cells separated from umbilical cord blood by density gradient centrifugation were cultured with orientated induction to differentiate the NSCs.The neuronal phenotype was identified using immunocytochemical methods.A total of 150 Sprague-Dawley rats were randomly divided into a sham-operation group,an HIBD group and an HIBD-NSCs group.Rats in the HIBD group and the HIBD-NSCs group were subject to ligation of the left carotid artery and then kept in a box under 8％ oxygen and 92％ nitrogen for 2.5 hours to establish the HIBD animal model.The artery was separated but not ligated in the sham operation group,which was not subjected to hypoxia.Twenty-four hours after the operation,the cultivated NSCs were transplanted by caudal vein injection into the rats in the HIBD-NSCs group.Rats were then sacrificed on the 3rd,7th,14th,21st and 28th days after the operation.Foxg1 gene expression in the SGZ was examined using in-situ hybridization methods.Results The number of Nestin-positive cells peaked on the 6th day of cultivation and then decreased by the 9th day.The Foxg1 gene was expressed in the SGZs of each group.The expression increased by the 3rd day after surgery in the HIBD and HIBD-NSCs groups,and peaked on 7th day after the operation,then declined gradually.The average expression level of Foxg1 in the HIBD group was significantly lower than that in the HIBD-NSCs group on the 7th day and thereafter.Conclusions Human umbilical cord blood mesenchymal stem cells can be induced and differentiated into neural stem cells.Foxg1 genes can still be present in the SGZ after birth.HIBD can induce the expression of Foxg1 genes.Transplanting NSCs can promote the expression of Foxg1 genes and improve morphological and functional recovery after HIBD,at least in neonatal rats.
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Objective To evaluate clinical features of three different gallbladder projective lesions. Methods Statistic analysis was performed for clinical data obtained from 325 cases of gallbladder projective lesions. Results There were 308 patients of benign lesions, including cholesterol polyps in 278 cases, other polyps in 5 cases, and adenomas of 25 cases. Malignant lesions was found in 17 cases, including 2 adenomas with malignant changes and 15 adenocarcinomas. 36. 0% and 32. 0% patients with benign polyps and adenomas were over 50 years of age, while 82.4% patients with adenocarcinomas were over 50 years. Average size of benign polyps, adenomas and adenocarcinomas were 8. 5 mm, 11.7 mm and 31.1 mm in diameter. 1.5% of benign polyps, 0% of adenomas and 52. 9% of adenocarcinomas were of low echo. 42.9% of benign polyps, 68.0% of adenomas and 100% of adenocarcinomas were single polyp. Conclusion The nature of gallbladder projective lesions can be suggested by patient age, size, number, and intensity of gallbladder projective lesions under B-ultrasound.