Objective To establish the method of identifying MRSA with Taqman-fluorescence quantitative PCR basing on mecA/nuc/fem B three gene combined detecting.Methods Taking the coagulase positive MRSA,which isolated from the clinical samples and confirmed by VITEK 2 compact microbial analyzer,as the research obj ect,designed mecA/nuc/fem B specific PCR primers and Taqman fluorescent probe by bio-software PrimerPremier 5 and Designer Beacon 7,FAM,HEX and ROX markers were used to label the fluorescent probe at 5’,and the end of 3’was labeled with BHQ1,detected by fluo-rescence quantitative PCR instrment.Results ①1 g/dl gel electrophoresis results showed that the primer’s specificity of mec A/nuc/fem B were good,and molecular weight of the amplification band consistent with the expected molecular weight and no non-specific amplification band.②Three genes were obtained specific amplification in a single tube single channel and single tube multiple channel detection in PCR,and the three gene amplification effect in a single tube single tube single chan-nel and multichannel PCR similar.Conclusion Successfully established a method of multi channel Taqman-probe fluores-cence quantitative PCR identification of MRSA,mec A/nuc/fem B combined detection can effectively differentiate coagulase negative and positive MRSA,improve the accuracy of identification.

Objective To evaluate the therapeutic effects of liver transplantation (LT) for cholangiocarcinoma(CC)and analyze the prognostic factors.Methods From December 2001 to December 2006,234 patients receiving LT for hepatic carcinoma in our institute were enrolled as a basis of comparative study for 6 CC patients undergoing LT during the same period.Results These 6 patients were followed-up from 1 to 56 months.Five patients died and one recurred.The 0.5-,1-and 2-year patient cumulative survival rates were 4/6,3/6 and 1/6,respectively.The 0.5-,1-and 2-year tumor-free survival rates were 3/6,2/6 and 1/6,respectively.The average patient or tumor-free survival time were both(14±4) months.Conclusion The prognosis of cholangioearcinoma patients after LT iS poor.

Objective To summarize the clinical experience in liver retransplantation. Methods The clinical data of 24 patients receiving liver retransplantation 28 times in this hospital were retrospectively analyzed and discussed with relevant literature. Results Among the 880 consecutive liver transplantations, 28(3.18%) had liver retransplantation. The causes of liver retransplantation were biliary complications ( 16 cases, 57. 1%), carcinoma recurrece (6 cases, 21. 4%), hepatic artery thrombosis (4 cases, 14. 3%), chronic rejection (1 case, 3. 6%), primary nonfunction (1 case, 3.6%). Thirteen patients among the 24 were discharged healthy and were followed up for 51days to 67months. Eleven patients died. Three of them died of hemorrhagic shock, 2 of septic shock, 2 of hepatocellular carcinoma recurrence, 2 of cardiovascular system complication, 1of nervous system complication, and 1 of hepatic artery thrombosis. Conclusion Liver retransplantation can effectively save patients with graft failure. Proper indication, optimal operating time, improvement of operative skills,and appropriate treatment during the perioperative period are very important for promoting the rate of successful liver retransplantation.

Objective To evaluate the role of transient receptor potential vanillic acid subtype 1 (TRPV1)/calcitonin gene-related peptide (CGRP) signaling pathway in lidocaine postconditioning-induced reduction of myocardial ischemia-reperfusion (I/R) injury in rats.Methods Forty healthy male SpragueDawley rats,aged 3 months,weighing 250-300 g,were divided into 5 groups (n=8 each) using a random number table method:sham operation group (group Sham),group I/R,lidocaine postconditioning group (group LP),lidocaine postconditioning plus CGRP8-37 group (group LP+CGRP8-37),and lidocaine postconditioning plus capsazepine group (group LP+Capz).Myocardial ischemia was induced by ligating the anterior descending branch of left coronary artery for 30 min,followed by 120-min repeRFusion in anesthetized rats.Lidocaine 2 mg/kg was injected via the tail vein at 5 min before reperfusion in group LP.In group LP + CGRP8-37,lidocaine was intravenously injected,and CGRP receptor selective antagonist CGRP8-37 2 mg/kg was intravenously injected at the same time.In group LP+Capz,lidocaine was injected intravenously,and TRPV1 blocker capsazepine 3 mg/kg was injected intravenously at the same time.A catheter was retrogradely implanted to the left ventricle,and heart rate (HR),left ventricular systolic pressure (LVSP),left ventricular end-diastolic pressure (LVEDP),and the maximum rate of increase or decrease in left ventricular pressure (±dp/dtmax) were continuously monitored and recorded.Blood samples were collected from the carotid artery at 120 min of reperfusion for determination of cardiac troponin I (cTnI),myoglobin (Myo) and creatine kinase-MB (CK-MB) concentrations in serum.Rats were then sacrificed for determination of myocardial infarct size.Results Compared with Sham group,the serum concentrations of cTnI,Myo and CK-MB were significantly increased,LVSP,+dP/dtmax and HR were decreased,and LVEDP and-dP/dtmax were increased in I/R group and LP group (P＜0.05).Compared with group I/R,the serum concentrations of cTnI,Myo and CK-MB and myocardial infarct size were significantly decreased,LVSP and +dP/dtmax were increased,and LVEDP,-dP/dtmax and HR were decreased in group LP (P＜0.05),and no significant change was found in the parameters mentioned above in group LP+ CGRP8-37 and group LP+Capz (P＞0.05).Compared with group LP,the serum concentrations of cTnI and Myo,myocardial infarct size and LVEDP were significantly increased,and + dP/dtmax was decreased in group LP+CGRP8-37,and the serum concentrations of cTnI and Myo and myocardial infarct size were significantly increased,LVSP and +dP/dtmax were decreased,and LVEDP was increased in group I/R+Lido+Capz (P＜0.05).Conclusion The mechanism by which lidocaine postconditioning mitigates myocardial I/R injury is related to activating TRPV1/CGRP signaling pathway in rats.

Objective To investigate the surgical options for the management of portal vein thrombosis (PVT) during liver transplantation and its impact on the outcome of patients. Methods 773 cases of liver transplantation were analyzed retrospectively. PVT occurred in 107 patients, inclu-ding 59 of grade Ⅰ ,33 of grade Ⅱ, 12 of grade Ⅲ and 3 of grade Ⅳ. Simple thrombectomy or thrombus-extraction was performed in grade Ⅰ and Ⅱ. 12 patients with grade Ⅲ received thrombus-extraction or using the donor iliac vein to act as a bridge between the donor portal vein and host superior mesenteric vein. Two cases of grade Ⅳ received a modified cavo-portal hemitransposition and one case received portal-vena coronaria varication anastomosis. Results Liver function had a good recover and the perio-perative mortality is 4. 3% in grade Ⅰ and Ⅱ. In grade Ⅲ , 5 cases received thrombus-extraction had a normal liver function after transplantation and had no died. 2 cases among the other 7 cases using por-tal vein reconstruction had bad liver function and died. The liver function recovered well after trans-plantation and there was no died in grade Ⅳ. Conclusions PVT is not a contraindication for liver transplantation. Good results can be obtained by applying reasonable operative procedures individually.

Objective To investigate the predictive value of Rotterdam CT score combined with serum soluble cluster of differentiation antigen 40 ligand (sCD40L) and fibulin-5 for prognosis of patients with severe traumatic brain injury (sTBI). Methods A total of 186 sTBI patients were divided into good prognosis group (n=104) and poor prognosis group (n=82). CT images were analyzed and Rotterdam CT scores were obtained; the enzyme-linked immunosorbent assay (ELISA) was used to detect serum sCD40L and fibulin-5 levels on the 1st, 3rd and 7th day of admission; the Spearman analysis was used to assess the correlations of serum sCD40L and fibulin-5 levels on the first day of admission with the Rotterdam CT score and Glasgow Coma Scale (GCS) score; the Multivariate Logistic regression analysis was conducted to explore the risk factors for poor prognosis in sTBI patients; the receiver operating characteristic (ROC) curve was performed to analyze the predictive value of Rotterdam CT score combined with serum sCD40L and fibulin-5 levels on the first day of admission for prognosis of sTBI patients. Results On the 1st, 3rd and 7th day of admission, compared with the good prognosis group, the serum levels of sCD40L and fibulin-5 in the poor prognosis group were significantly elevated (P < 0.05); serum sCD40L and fibulin-5 levels were negatively correlated with GCS scores (r=-0.505, -0.421, P < 0.05) and positively correlated with Rotterdam CT score (r=0.495, 0.397, P < 0.05); sCD40L (OR=2.768, 95%CI, 1.537 to 4.983) and fibulin-5 (OR=2.539, 95%CI, 1.301 to 4.953) were independent risk factors for poor prognosis in sTBI patients (P < 0.001); the area under the curve (AUC) of Rotterdam CT score combined with serum sCD40L and fibulin-5 levels on the first day of admission for predicting poor prognosis in sTBI patients was 0.969 (95%CI, 0.933 to 0.989), with a sensitivity of 86.59% and a specificity of 94.23%; the diagnostic performance of Rotterdam CT score combined with sCD40L and fibulin-5 was superior to that of each individual indicator (Z=4.233, 4.274, 4.433, P < 0.001); the Bootstrap internal validation results showed that the predictive performance curve of the combined prediction model was consistent with the actual clinical occurrence curve. Conclusion Rotterdam CT score combined with serum sCD40L and fibulin-5 has certain predictive value for poor prognosis in sTBI patients.

The clinical data of 67 patients underwent orthotopic liver transplantation, including 4 cases of early intra abdominal hemorrhage after orthotopic liver transplantation, were analyzed retrospectively.Anastomotic liver artery hemorrhage was found in 2 cases, hemorrhage of IVC in 1 case and hemorrhage of right adrenal gland in 1 case. All the 4 patients were correctly dignosed and successfully treated by operation in time. It suggests that the early intra abdominal hemorrhage after orthotopic liver transplantation is mostly due to surgical technique. The improvement of surgical technique,correct diagnosis and timely operation are very important for the treatment of intra abdominal hemorrhage.

Objective To investigate some improvements in the surgical techniques of adult-to-adult living donor liver transplantation( A-A LDLT) without the middle hepatic vein(MHV) for hepat-ic vein reconstruction. Methods The retrospective analysis was made on the clinical data of 11 recipi-ents who underwent the operation in A-A LDLT including the hepatic vein reconstructed in right liver lobe without MHV from June 2007 to January 2008. The key techniques included reconstructing out-flow of graft on shaping the tips of vena cava and right hepatic veins, cadaveric vein allografts stored in 4℃ UW solution within 7d being used for significant-sized hepatic vein reconstruction such as tributa-ries of the middle hepatic vein from V5, V8 and right inferior hepatic vein. Results 10 cases success-fully underwent reconstruction of outflow of graft on shaping the tips of vena cava and right hepatic veins and the outflow reconstruction ratio of V5, V8 and right inferior hepatic vein was 81. 8% (9/11), 7 one-vein reconstruction, 1 two-vein reconstruction and 1 three-vein reconstruction. 1 recipient died of renal failure and pulmonary infection 14 days after operation without venous outflow obstruc-tion. Doppler ultrasonography showed no thrombosis and the blood flowed smoothly in the right he-patic vein of other 8 recipients during the 9th to 15th mouth of follow-up. The cumulative patency rates of these 8 survivals for interposition vein grafts were 100% (11/11), 72. 7 %(8/11), 54. 5%(6/11) and 36. 5%(4/11) in 1, 3, 6 and 9 mouths, respectively. The regeneration of paramedian sectors was equivalent. Conclusion Shaping the tips of vena cava and right hepatic veins and using cadaveric vein allografts in adult-to-adult right lobe living donor liver transplantation for hepatic vein reconstruc-tion are both safe,simple and effective methods.This approach can be recommended.

Objective: To evaluate the effects of ketamine anesthesia on proteome in hippocampus in aged rats. Methods: Thirty healthy male Wistar rats, aged 20 months, weighing 560-610 g, were divided into 2 groups (n=15 each) using a random number table method: control group (group C) and ketamine group (group K). In group K, ketamine 80 mg/kg was intraperitoneally injected, additional 1/2 initial dose was given when the righting reflex was recovered, and anesthesia was maintained for 3 h. Morris water maze test was performed starting from 1st day after the end of anesthesia.Five rats were selected at days 1 and 7 after the end of anesthesia and sacrificed, and hippocampal tissues were obtained to extract proteins.Proteins extracted from rat hippocampi were identified by 2-dimensional electrophoresis (2-DE). The differentially expressed proteins were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and biological information system. Results: Compared with group C, the escape latency and total swimming distance to find the submerged platform in Morris water maze at the 1st day after anesthesia were significantly prolonged in group K (P<0.05 or 0.01). The MALDI-TOF-MS analysis showed that there were 21 differentially expressed proteins at 1st day after ketamine anesthesia, of which 6 proteins (involving maintenance of intracellular protein homeostasis, energy metabolism, etc.) presented with up-regulated expression and 15 proteins (involving synaptic vesicle transport efficiency, synaptic structural and functional plasticity, maintenance of intracellular protein homeostasis, NMDA-mediated Ca²⁺ signal transport, energy metabolism, etc.) presented with down-regulated expression.There were 8 differentially expressed proteins at 7th day, including 3 proteins with up-regulated expression and 5 proteins with down-regulated expression (P<0.05). Conclusion Ketamine anesthesia can induce 21 differentially expressed proteins in hippocampi of aged rats, involving synaptic vesicle transport efficiency, synaptic structural and functional plasticity, intracellular protein homeostasis, NMDA-mediated Ca²⁺ signal transport, energy metabolism, and etc.which may be involved in the mechanism of ketamine-induced temporary cognitive dysfunction.

Objective:To investigate whether ventricular non-excitatory electrical stimulation(NES)preconditioning can protect myocardial ischemia-reperfusion(IR)injury in aged mice by regulating cardiac calcitonin gene-related peptide(CGRP).Methods:Male C57BL/6J mice were randomly divided into 3 groups, (1)the Sham group, receiving sham operation group(n=6); (2)the IR group, receiving ligation of coronary artery to induce myocardial ischemia for 45 min and reperfusion for 120 min(n=13); (3)the NES group, IR model receiving NES from 15 min before IR to the end of reperfusion(n=13). Infarct size was detected by staining with 2, 3, 5-triphenyltetrazolium chloride.The level of serum cTnI and expression of myocardial CGRP were measured by enzyme linked immunosorbent assay and quantitative real time polymerase chain reaction.Results:Compared with the IR group, the infarct sizes were significantly lower in NES group[(38.17±4.36)% vs.(45.33±5.68)%, P<0.05]. Besides, the IR and NES groups were associated with significantly increased levels of serum cTnI[(10.89±2.14)μg/L, (7.03±1.79)μg/L vs.(3.92±0.47)μg/L, P<0.001], myocardial CGRP protein[(26.33±4.55)μg/kg, (19.67±5.79)μg/kg vs.(17.00±2.90)μg/kg, P<0.01], CGRP mRNA[(1.40±0.20), (2.20±0.75) vs.(1.05±0.10), P<0.01]compared with the Sham group.Furthermore, the NES group was associated with markedly decreased levels of serum cTnI and myocardial CGRP protein, and increased level of CGRP mRNA compared with the IR group(all P<0.05). Conclusion:NES may protect myocardium IR injury by regulating endogenous CGRP expression in aged mice.