We designed two pairs of primers and their corresponding TaqMan probes according to gH, gE gene of PRV. By optimizing the probe's concentration, Mg2+ concentration, primers concentration and sample DNA extraction, real-time fluorescent quantitative PCR (FQ-PCR) which can quickly identity field virus and vaccine virus of PRV was established. According to our results, the dynamic range of the FQ-PCR assay is between 10 x 10(1) copies/microL and 10 x l0(8) copies/microL, and the detection limit of FQ-PCR is 1.0 x 10(1) copies/microL, which is 100 fold higher than that of conventional PCR. We detected 60 doubtful tissue samples using the FQ-PCR assay, serum neutralization and conventional PCR. In conclusion, the FQ-PCR method is rapid, sensitive, specific and accurate, and can be used to detect field strains of PRV rapidly. The closed-tube format of the assay minimized the risk of contamination of subsequent reaction and the assay can be performed in 2 h or less. Development of real-time quantitative PCR provides the basis for the early and rapid detection and analyzing quantitatively the infectious degree of PRV.
Animals ; Fluorescent Dyes ; Herpesvirus 1, Suid ; genetics ; isolation & purification ; Polymerase Chain Reaction ; methods ; Pseudorabies ; diagnosis ; prevention & control ; virology ; Pseudorabies Vaccines ; immunology ; isolation & purification ; Swine

Objective To examine the benefits of ECMO for potential organ donors with hemodynamic instability after brain death.Methods Three brain-dead potential donors who presented with hemodynamic instability despite maximal medical management,finished a declaration of brain death,that were supported by extracorporeal circulation membrane oxygenation (ECMO).Results Donor organs,including six kidneys,and two livers,were harvested from the three donors under ECMO support,leading to 8 successful transplantations.The organs functioned well and the recipients made full recoveries.Conclusion Our experience indicates that ECMO allows for the maintenance of abdominal organ tissue perfusion without warm ischemia before organ procurement,providing sufficient time for safe organ donation procedures and reducing the risk of unpredictable cardiac arrest that could result in the donor death and graft loss.

Objective To summarize the short-term results of simultaneous pancreas-kidney transplantation (SPK) at a single center in China.Methods SPK was performed on 12 consecutive patients from Jan.2010 to July 2014.All patients had long-standing insulin-dependent diabetes mellitus (IDDM) and subsequent renal failure.Bladder drainage (BD) of exocrine secretion was used in the 10 cases and enteric drainage (ED) in 2 patients.The patients were treated with quadruple therapy,which included ATG or anti-CD25 monoclonal antibody induction therapy,prednisone,tacrolimus and mycophenolat-mofetil (MMF).Results The SPK was performed successfully in 10 cases.One patient accepted re-pancreas transplantation due to necrotizing pancreatitis.One patient suffered hemorrhage of bladder,accepted 3 times of embolization therapy and died due to lung infection.Ten patients achieved excellent renal function and euglycemia,and no further insulin treatment was given in 9.5 ± 4.2 days posttransplant.Fasting plasma glucose returned to normal in 14.2 ± 5.1 days.Serum creatinine returned to normal in 10.4 ± 6.5 days.The mean hospital stay was 21.4 ± 7.3 days.One biopsy-proven renal rejection episodes occurred in 14 days postoperation.Main complications included wound infections on the side of pancreatic graft,lymphorrhagia,tacrolimus toxicity and urinary tract infection.Conclusion SPK is an effective therapy of ESRD.Donated graft protection system foundation,refinement and individualized treatment posttransplantion may be the key factors for successful SPK.

BACKGROUND:Delayed graft function (DGF) occurs frequently in kidney transplants from donation after cardiac death if creatinine level is high in kidney recipients. OBJECTIVE:To analyze the clinical effects of renal transplantation with kidneys from donors dying of cardiac death in organophosphate poisoning. METHODS:Data were col ected from kidney transplants from two donors dying of cardiac death in organophosphate poisoning. After some donor maintenance, donor organ were obtained and perfused with impulse type machine. Recipients were treated with intervention of immunity induction, anti-rejection drugs and infection prevention drugs during and after renal transplantation. Pathological data of donor kidney zero needle biopsy, DGF after kidney transplantation, complication rate (such as acute rejection), renal al ograft recovery situation, the survival rate of recipients and kidney transplants were col ected and analyzed. RESULTS AND CONCLUSION:Needle biopsy results from four donor kidneys showed that glomerular morphology was normal, but there were edema and degeneration in kidney tubules in some degree. Donor DGF rate was 75%(3/4), acute rejection rate was 0%(0/4), perioperative period donor kidney and recipient survival rate were 100%(4/4). Al recipients showed a good result of transplanted kidney, their creatinine and urea nitrogen were at low level, and had no proteinuria. One recipient died of severe pulmonary infection 4 months after surgery. For some organophosphate poisoning donors dying of cardiac death, donor kidney quality can be improved by suitable donor maintenance and high-quality donor kidney preservation using machine perfusion. Kidney transplants from donors dying of cardiac death in organophosphate poisoning who receive the maintenance of organ function may be a promising candidate for renal transplantation due to a severe lack of kidney donor sources.

Objective To compare the therapeutic effects of orthotopic injection and tail vein injection of human amniotic mesenchymal stem cells(hAMSCs)on histological restoration and neurological functions of rats with spinal cord injury. Methods Transected spinal cord injury model in rats was established by transplanting DAPI prelabelled hAMSCs one week after injury.BBB scores were used to evaluate the hindlimb movement of rats. The histological patterns.and morphology of medullary sheath of spinal cord were observed. Results BBB scores in the orthotopic injection group and tail vein injection group were increased gradually from one to six week after hAM-SCs transplantation and reached 6.5 ± 0.5 and 7.12 ± 1.61 respectively 6 weeks after cell transplantation,higher than that of the control group(both P < 0.01). However,there was no statistical significance between the two groups.Histological results indicated that the repair of injured tissue in the orthotopic injection group and tail vein injection group were both better than that in the control group,and there were more vesica and loosened layers forming in the injured spinal cord of rats in the PBS control group as compared with the orthotopic and tail vein transplantation group. Conclusion hAMSCs transplantation through tail vein injection could promote histological restoration and neurological regeneration of rats with spinal cord injury,which has the similar therapeutic effects with hAMSCs orthotopic transplantation.

AIM:To explore the effect and mechanism of the interfering Gal-1 on epithelial-mesenchymal transition(EMT),migration and proliferation in MDA-MB-231 cells via transforming growth factor-β(TGF-β)pathway.METHODS:The stable cell lines(shGal-1)which Gal-1 expression were inhibited completely and their control cell lines were used as experimental cells.Western blot assay was used to detect the effects of shGal-1 on EMT process of MDA-MB-231 cells after TGF-β treatment;The effect of shGal-1 on cell migration and invasion after TGF-β treatment was verified by cell scratch and transwell test;The effect of shGal-1 on the TGF-β pathway related proteins were detected by western blot;Finally,the effect of shGal-1 on cell proliferation was detected by MTT and western blot.RESULTS:shGal-1 inhib-ited TGF-β-mediated EMT in MDA-MB-231 cells and regulated phosphorylation of pathway signaling molecules(ERK,AKT and GSK3β);shGal-1 could inhibit the proliferation of MDA-MB-231 cells.CONCLUSION:shGal-1 can inhibit the TGF-β-mediated EMT,migration and proliferation of MDA-MB-231 cells.

Objective To evaluate the effect of conversion from mycophenolic acid (MPA) to mizoribine (MZR) in renal transplant recipients with gastrointestinal tract (GI) symptoms.Methods A total of 355 renal transplant recipients with GI symptoms caused by MPA administration were enrolled from April 2015 to March 2017 in 25 different renal transplant centers in China.The symptomatic improvement of GI before (baseline) and after conversion to MZR (1,2,4 weeks) was assessed by each item of GI symptoms indication.In addition,the efficacy and safety of the conversion therapy during 12 months were determined.Results Patients showed improvement in GI symptoms including diarrhea,abdominal pain,abdominal distention and stomachache after conversion to MZR 1,2,4 weeks (P＜0.05).In patients with different severity of diarrhea,conversion to MZR therapy significantly improved diarrhea (P＜0.05).During 12 months,no patient experienced clinical immune rejection.We did not observe any infections,leucopenia and other serious side effects.Conclusion MZR could markedly improve GI symptoms caused by MPA administration in renal transplant recipients.

Objective By analyzing the perioperative management in our hospital to explore the clinical effect and safety of single kidney transplantation from deceased juveniles' donors.Methods We retrospectively analyze 86 cases of kidney transplantations from deceased juveniles' donors in our hospital from 2007 December to 2015 August.Results The success rate of the operations was 100％.The postoperative complications occurred as fellows:7 cases of acute rejection (8.14％);10 cases of drug intoxication (11.62％);21 cases of DGF (24.44％),4 cases of leakage of urine (4.65％),7 cases of lung infection (8.14％).Two cases (2.32％) died after the operation because of serious lung infection,and by corresponding treatment 47 cases recovered after 2-4 weeks.The creatinine level in 37 cases without any complications was 131.88 ± 44.20 μmol/L during discharge.Conclusion With strict selection,the organ from a deceased juvenile donor is safe and practicable.

Myocardial injury is a pathological change of myocardium caused by many factors,which can lead to the decline of cardiac function and the occurrence of cardiovascular events.Astragaloside Ⅳ is one of the main pharmacological components in Astragali Radix,which plays an anti-myocardial injury role by regulating various signaling pathways.This article reviewed the anti-myocardial injury mechanism of astragaloside Ⅳ from five aspects:inhibition of oxidative stress,inhibition of apoptosis,anti-myocardial fibrosis,improvement of myocardial energy metabolism and inhibition of myocardium inflammation,in order to provide reference for the mechanism research and clinical application of astragaloside Ⅳ in the prevention and treatment of myocardial injury.

Objective:To evaluate the efficacy and safety of continuous infusion of low-dose intravenous (Ⅳ) heparin during perioperative period of simultaneous pancreas-kidney (SPK) transplantation for donation after citizen death (DCD) donor to prevent pancreatic thrombosis post-transplantation.Methods:From January 2015 to August 2019, 46 DCD donors undergoing SPK were divided into retrospective cohort groups 1 ( n=27) and 2 ( n=19). Group 1 received aspirin enteric-coated tablets only at Day 1 post-SPK. In Group 2, 5-7 days of continuous infusion of heparin 260 IU per hour at Day 1 post-SPK was followed by a daily intake of aspirin enteric-coated tablets of 100 mg. Incidence of thrombus, recovery of graft function and adverse reactions of anticoagulant therapy were observed. Results:Thrombosis occurred in (5.3%, 1/19 vs 14.8%, 4/27) in heparin and non-heparin groups. Thrombosis and graft loss were significantly lower in heparin group than those in non-heparin group ( P<0.05). Conclusions:Continuous infusion of low-dose heparin vein is effective and safe in preventing thrombosis after SPK transplantation.