Objective To compare the early renal function recovery after kidney transplant from donation after cardiac death (DCD) and brain death (DBD).Methods The Medline (1950-2011),Embase and Cochrane library database were searched and supplemented by review of conference proceedings and publication bibliographies.All original single institution studies reporting outcomes for DCD and DBD kidney transplant recipients were considered.Odds ratios (OR) and 95％ confidence intervals (CI) based on random effects models were calculated.Results Nine publications,all cohort studies,involving 2049 DCD and 5498 DBD recipients,were included.DCD recipients had 7.24 times increased odds of DGF (OR=7.24,95％ CI =3.86-13.58),and 4.97 times increased odds of PNF (95％ CI =3.77-6.55).Conclusion DCD renal transplantation is associated with higher risks of DGF and PNF.

AIM: To study the effect of microelement powder (MP) on membrane potential of vascular endothelial and smooth muscle cells of rats in order to elucidate the mechanism of microcirculation improvement by MP. METHODS: Cultured pulmonary vascular endothelial cells (EC) and aortic smooth muscle cells (SMC) of rats and detecting the changes of cellular membrane potentials by using potential-sensitive fluorescent probe and laser jet confocal microscope. RESULTS: MP hyperpolarized SMCs significantly. Glybenclamide (2 μmol/L), a blocker of KATP channel, which had no effect on membrane potential of SMCs, reversed the hyperpolarization of MP completely; MP hyperpolarized ECs slightly, but the effect was unaffected by glybenclamide. CONCLUSION: MP hyperpolarizes SMCs by activating KATP channels and leads to dilation of microvessels and improvement of microcirculation.

AIM: To study the effect of microelement powder (MP) on membrane potential of vascular endothelial and smooth muscle cells of rats in order to elucidate the mechanism of microcirculation improvement by MP. METHODS: Cultured pulmonary vascular endothelial cells (EC) and aortic smooth muscle cells (SMC) of rats and detecting the changes of cellular membrane potentials by using potential-sensitive fluorescent probe and laser jet confocal microscope. RESULTS: MP hyperpolarized SMCs significantly. Glybenclamide (2 ?mol/L), a blocker of K ATP channel, which had no effect on membrane potential of SMCs, reversed the hyperpolarization of MP completely; MP hyperpolarized ECs slightly, but the effect was unaffected by glybenclamide. CONCLUSION: MP hyperpolarizes SMCs by activating K ATP channels and leads to dilation of microvessels and improvement of microcirculation.

Objective:Human adipose-derived stem cells (hASCs)are a highly attractive source in bone tissue engineering.To generate a luciferase reporter system that could be used to quantitatively and rapidly examine osteogenic differentiation potential of human adipose-derived stem cells (hASCs ) in vitro,and eventually make it possible to monitor the osteogenic differentiation of transplanted cells in vi-vo.Methods:The genomic DNA harboring promotor regions of osteocalcin and DNA sequences encoding luciferase genes were amplified by PCR and cloned into the pLVX-pTRE-puro vector to generate the OCpro-Luc-Puro construct.Then,the OCpro-Luc-Puro construct together with three assistant vectors:pM-DLg/pRRE,pRSV-REV,and pVSVG,were transiently transfected into HEK293T cells followed by viral supernatants collection,filtration and concentration.Next,the hASCs stably expressing luciferase repor-ter gene driven by osteocalcin promotor were created with the lentivirus carrying OCpro-Luc-Puro cassette under puromycin selection.The OCpro-Luc-hASCs were then cultured in the absence or presence of osteo-genic differentiation medium.On the 7th and 1 4th days,after osteogenic induction,cellular extracts were collected and analyzed by luciferase reporter assay.Meanwhile,alizarin red staining and quantification as well as quantitative reverse transcription PCR (qRT-PCR)analysis of osteogenic associated genes osteo-calcin (OC),runt-related transcription factor 2 (Runx2)and alkaline phosphatase (ALP)were used to assess the osteogenic differentiation ability of OCpro-Luc-hASCs.Results:OCpro-Luc-Puro plasmid and OCpro-Luc-hASCs were successfully generated.On the 7th and 1 4th days after osteogenic induction,the luciferase activity of the cellular extracts from OCpro-Luc-hASCs was dramatically increased.Consistently, the extracellular matrix mineralization,as shown by Alizarin red S (ARS)staining and quantification was also markedly intensified and a marked increase of the mRNA expression levels of OC,Runx2 and ALP, although to variable extent,was observed upon osteogenic differentiation.These results indicated that the observations from traditional experiments examining hASCs osteogenic differentiation were largely in agreement with that of our luciferase reporter assay in OCpro-Luc-hASCs.Conclusion:We established a luciferase reporter system that could be used to rapidly,quantitatively and specifically determine osteo-genic differentiation ability of hASCs.Comparing with the traditional time-consuming methods,the system we generated here was highly effective.This system not only can be used to examine ostogenic differentia-tion of hASCs in a high throughput manner,but also provides a way to monitor ostogenic differentiation of cells in vivo.

Graphene oxide (GO) modified stationary phase for open tubular capillary (3 m × 25 μm i. d. ) of liquid chromatography ( OT-CLC) was fabricated by coating GO sheets onto poly ( vinylbenzyl chloride-divinylbenzene) porous composite layer via covalent coupling, which was prepared by in-situ polymerization. Scanning electron microscopy ( SEM), Raman spectroscopy and transmission electron microscopy ( TEM) were used to characterize the structure of the stationary phase. The results demonstrated that poly(vinylbenzyl chloride-divinylbenzene) porous composite layer had decentralized globular structure, and the GO sheets covered on the porous layer homogenously. The phase ratio and sample capacity were greatly improved due to the spherical polymer layer and the coverage of GO. Thus, alkyl benzenes, neutral polycyclic aromatic hydrocarbons, acidic and basic compounds could be well separated by using acetonitrile-water as eluent, while four nucleobases were completely separated by using acetonitrile-0. 02 mol/ L ammonium acetate as mobile phase. The run-to-run, day-to-day, and column-to-column reproducibilities were evaluated by calculating the relative standard deviations (RSDs, n =6) of the retention time of o-phenylenediamine, aniline and 2, 4, 6-trifluoroaniline, respectively. These RSD values were all in the range of 0. 3% -2. 0% .

At present, the laser induced breakdown spectroscopy (LIBS) has already become an analysis method with great application value and bright prospects. It has been reported that the LIBS technology had been successfully applied in the field of clinical laboratory diagnostics such as the identification of pathogens, the diagnosis of malignant tumors, the identification of caries, the analysis of onychomycosis, the detection of toxic and harmful elements and other applications with positive results. The remote-LIBS technique provides feasibility for detecting pathogen which is with significance of the disease prevention and control. It is possible to apply LIBS technology to in vivo diagnostics in the future.

Objective To analyze the role of the introperative vessel Doppler sonographic evaluation of the hepatic artery and portal vein. Methods Intraoperative vessel Doppler sonograms of 116 patients were analyzed for peak systolic velocity of hepatic artery and blood flow of the portal vein.In patients having abnormal findings on sonography (peak systolic velocity of hepatic artery less than 30 cm/s, blood flow of the portal vein less than 800 ml/s), the vascular anastomoses were checked.Results Fourteen of 116 cases revealed less hepatic arterial peak systolic velocity than 30 cm/s. In 9 of the 14 cases, the hepatic arterial peak systolic velocity was normal after injection of 0. 5 % lidocaine into celic trunk root, and papaverine and 654-2 into artery, 3 of the 9 cases endured artery thrombosis. In the other 5 of the 14 cases, by-pass anastomoses were done, and the hepatic arterial peak systolic velocity was normal, and no hepatic arterial complication occurred. Five of 116 cases revealed less hepatic portal vein blood flow than 800 ml/rnin. 4 of the 5 cases revealed shunt between portal vein and vena cava. The blood flow was normal after ligation of the shunt, and thrombosis occurred in 1 case of the 4. The another 1 of the 5 cases was presented with portal vein thrombosis of grade m, and the blood flow remained lower than normal when side-to-side anastomosis was done after resection of thrombosis. Then vein by-pass of the superior mesenteric vein to portal vein with donor iliac vein was done, the blood flow became normal, and no complication occurred. Conclusions The vessel Doppler sonography during liver transplantation was of pivotal values in preventing and diagnosing vessel complications. For the patients with abnormal findings though intraoperative vessel Doppler sonography, the close monitoring should be done in order to find out vessel complication as

Objective Pancreas perfusion is an essential step in human islet isolation.To develop the new methods for introductal canulation,collagenase infusion and to observe their effects on islets isolation.Methods A total of 17 pancreases were digested from March 2005 to April 2010.The pancreases were distended by three methods:the standard method (n =3),the one-cannula method (n =11) and three-cannula method (n =3).In the standard group,the pancreases were completely cut into half at the mid-body.Two catheters were inserted into the main duct:one directed toward the tail and the other to the head.In the one-cannula method group,a long tube was inserted into the duct at the head,advancing to the tail In the three-cannula method group,pancreatic parenchyma was then minimally cut at the mid-body and three catheters were inserted into the main pancreatic duct:one at the head (the first catheter) and two at the mid-body,one toward the tail (the second catheter) and the other toward the head (the third catheter).The pancreases were digested by improved Ricordi technique.Ficoll continuous density/grads centrifuge method was performed to purify the islets.DTZ staining was adopted to identify islets and count islet equivalent (IEQ). AO/EB fluorescence examination was used to count active islet percentage.Static glucose stimulating test (SGS) in vitro was designed to estimate islet function and calculate SI.Results The distension volume of the threecannula method group was 1.24 rnl/g pancreas,and higher than the other groups (for the standard group:0.71 ml/g pancreas; for one-cannula method group:0.96 ml/g pancreas,P＜0.05).The yield of islet in the three-cannula method group and the one-cannula method group was 2514 and 2270 IEQ/g,which was significantly more than that in the standard group (1914 IEQ/g pancreas,P＜0.05).The purity and viability of the islets were 74 ％/79.3 ％,75.6 ％/79.4 ％ and 78.3 ％/84.0 ％ respectively in the three groups with the difference being not significant among the groups.SI in the one cannula method group (4.74) and the three-cannula method group (5.27) was significantly higher than that in the standard group (3.46).ConclusionThe three-cannula method improved collagenase infusion and the islet yields.

Objective To summarize our experience in the liver transplantation from the donation after cardiac death (DCD).Methods The livers from three DCD donors (2 cases of brain trauma and 1 case of cerebral hemorrhage) were harvested according to the Guidelines for Donation after Cardiac Death in China.These grafts were orthotopically transplanted into three recipients including 2 cases of decompensative hepatic cirrhosis and 1 case of primary liver cancer.The warm ischemic time ranged from 7.5 to 10 min and the cold ischemic time was 4.5,8.2 and 6.5 h respectively.Postoperative immunosuppressive regimens included prednisone,FK506 and mycophenolate mofetil (MMF).Antibiotics and anticoagulatants were used accordingly.Results All of the 3 recipients obtained normal liver function within 3 weeks since the grafts were implanted without PNF,thrombosis and rejection.No postoperative complications occurred in 3 recipients during the follow-up period of 2 to 9 months with normal liver function.Conclusion The liver transplant from DCD donor showed good results in our center.Chinese group Ⅲ of DCD donor,UW score above the middle level and the short warm ischemic time are three keys ensuring the success of the liver transplant from DCD donors.

Objective To summarize the clinical experience of harvesting and using the kidneys from donation after cardiac death (DCD) donors.Methods Fourteen kidney transplantations were successfully performed on 14 patients with end-stage renal diseases.The kidneys were harvested from 7 volunteer donors (age 30～53 years) diagnosed with cardiac death,who were scored 19～23according to the University of Wisconsin donation after cardiac death evaluation.Primary diseases of the donors were cerebral hemorrhage,brain injury,ischemic cerebral vascular disease and brain tumor.Warm ischemia time ranged from 5 to 45 min,and cold ischemia time was 4.5 ～ 12.5 h.Results After transplantation,three patients had delayed graft function (DGF),one had primary non-function (PNF),and two patients developed acute rejection.In the patient with PNF,the transplanted kidney was removed one day after operation and the patient went back to hemodialysis.One patient with DGF was still in recovery with serum creatine 149 μmnol/L (within 3 months after operation).The above two cases both utilized the kidneys with 45 min of warm ischemia time.The rest 12 patients were discharged with normal renal function.Conclusion Under the condition of our country,kidneys strictly harvested from DCD donors can be used as one of the main sources of kidney grafts for kidney transplantation.