5 cases of orthotopic live transplantation (OLTX) were performed in unresectable hepatic cellular carcinoma(3),Wilson's disease(1)and hepatitis B(1).The donor livers were harvested by the technique of rapid multiple organ harvesting,flushed and preserved with UW solution.The veno-venous bypass was introduced during the anhepatic phase in the operation.The double or triple-drug immunosuppressive regimen was used and three episodes of acute rejection were controlled in two patients.One patient died of CMV infection and another,of upper gastrointestinal hemorrhage on the 92nd and 58th postoperative day respectively.The other three are still alive for 35,150 and 210 days.

Objective To establish a method to isolate,culture and identify cardiomyocytes from neonatal mouse,estimate the cardiomyocytes survival rate,and identify the purity of cardiomyocytes.Methods Neonatal C5 7 mouse heart(within 3 days)was di-gested using trypsin and collagenase.Cardiomyocytes survival rate was estimated by trypan blue staining,and cardiomyocytes purity was identified byα-actin immunofluorescence staining.Results Mouse cardiomyocytes could be successfully isolated and cultured using neonatal mouse within 3 days.Trypan blue staining showed cardiomyocytes survival rate was>95%,and cardiomyocytes pu-rity was more than 95% demonstrated byα-actin immunofluorescence staining.Conclusion Under the strict experimental condi-tions,mouse cardiomyocytes can be successfully isolated and cultured with high survival rate and high purity.

Objective To investigate the alteration of peripheral tissue`s temperature of the coagulation zone of microwave ablation in brain tissue, and to provide experimental evidence for clinical application. Methods Twelve canines were treated by microwave ablation in brain tissue. Each was ablated for 180 s with microwave output power of 20 W, 30 W, and 40 W. During the operation the peripheral temperature at the distance of 0.5 cm, 1.0 cm, 1.5 cm and 2.0 cm from the ablation center was recorded respectively. The ultrasound was performed 1 hour after the operation, and then the animals were executed and the microscopic changes of the ablation lesion were observed. Results Eleven canines suffered well for the ablation, while 1 presented abnormal respiration during the operation and died 2 hours later. During the operation, the temperature of the area 0.5 cm from the center rose signiifcantly, with the maximum temperature was (96.40±1.46)℃at the power of 20 W, and 100℃at the power of 30 W and 40 W. The temperature of the area 1.0 cm from the center rose faster, with the maximum temperatures at different powers all above the 46℃. The temperature of the area 1.5 cm from the center rose slower, with the maximum temperature below 46℃at the power of 20 W and 30 W and above 46℃at the power of 40 W. The maximum temperatures of the area 2.0 cm from the center at different powers were all below 46℃. The difference of the maximum temperature at different distances (1.0 cm, 1.5 cm, and 2.0 cm from the center) was signiifcant (F=776.78, 2640.64 and 3025.53, all P<0.05). The length and width of the ablation lesion as well as the area of edema increased with the power. At the power of 20 W, 30 W, and 40 W, the length of the ablation lesion was (29.3±1.8) mm, (32.7±2.1) mm and (34.2±2.4) mm, the width was (22.5±1.5) mm, (23.7±1.7) mm and (27.1±2.0) mm, and the width of the edema zone was (2.3±0.4) mm, (2.6±0.4) mm and (2.7±0.5) mm. The differences of the length and width of the ablation lesion at different powers were signiifcant (F=11.46, 14.49, both P<0.01). The difference of the edema area at different powers was insigniifcant (F=1.94, P=0.169). Conclusions Microwave ablation is a safe therapeutic modality. However, the shorter distance from the ablation center and greater ablation power give rise to larger ablation lesion, higher maximum temperature, and faster temperature increase. Therefore, 2.0 cm from the ablation center is a safe area.

Objective To investigate the effects of cold exposure with different time courses on cardiovascular function and skeletal muscle contraction in Sprague-Dawley (SD) rats.Methods The male SD rats were randomly divided into control,12-h cold,24-h cold,and 48-h cold groups.The rats in all cold group were maintained in 4 ℃ environment for indicated time and allowed free access to standard rat chow and water ad libitum.After the animal model was established,the heart,thoracic aorta,soleus,and extensor digitorum longus (EDL) were rapidly isolated from rats under anesthesia and then fixed into the respective organ perfusion system that was connected to physiological recorders and data acquisition system.Lanendorff heart persusion was used to detect some parameters of cardiac function,such as coronary flow (CF),mean left ventricular systolic pressure (mLVSP),mean left ventricular diastolic pressure (mLVDP) and maximum ascending and declining rate of left ventricular pressure (± dp/dt max) ; and then the heart was suffered from 30 min ischemia followed by 60 min reperfusion,the changes of above cardiac function indexes were recorded again.Organ bath method was employed to detect contraction response to potassium chloride (KCl) or phenylephrine (PE) and endothelium-dependent relation responses to acetylcholine (ACh) of aortic rings.The twitch tension,titanic tension,and fatigue test of soleus and EDL were examined by electrical stimulation to reflect contractive function of skeletal muscle.Results The coronary flow (CF) were significantly reduced in 24-h and 48-h cold groups compared to control group [(9.0 ± 1.7) ml/g and (9.8 ± 1.3) ml/g vs (11.6 ± 1.2) ml/g,P ＜ 0.05] ; 48-h cold exposure also showed significantly increased mLVSP (P ＜0.01) and mLVP (P ＜ 0.05) of rats.After ischemia-reperfusion,the heart re-beat time was shorter in all cold exposure groups (all P ＜ 0.01),mLVP and mLVDP were higher in 24-h and 48-h cold group than control group (all P ＜ 0.01).The contraction of aortic rings responses to 60 mmol/L KCl [(0.63 ±0.13) g and (0.79 ±0.11)g vs (1.28±0.17)g,P ＜0.01] and 1 μmol/L PE [(0.62 ±0.18)g and (0.68 ±0.13)g vs (1.25 ± 0.18)g,P ＜ 0.01] were decreased while the endothelium-dependent relaxation responses to low concentration ACh (3 × 10-8 ～ 3 × 10-7) were increased in 24-h and 48-h cold groups relative to control group (all P ＜ 0.05).The 48-h cold also reduced titanic tension of SOL and EDL in comparison with control [(4.14 ± 0.62) N/cm2 vs (5.50 ± 0.75) N/cm2,P ＜ 0.05 and (3.00 ± 0.57) N/ cm2 vs (4.70 ± 0.85) N/cm2,P ＜ 0.01].Conclusions 24 ～ 48 h cold exposure could reduce CF but enhance the tolerance of heart from rats to ischemia reperfusion injury and also decrease the contraction of aortic rings response to KCl or PE but increase the sensitive responses to ACh of aortic rings from rats.The 48-h cold could decrease the contraction of skeletal muscle elicited by electrical stimulation.

Objective To investigate the effects of fasting with different time courses on the contraction of skeletal muscle and cardiovascular function in Sprague-Dawley (SD) rats.Methods The healthy and male SD rats were randomly divided into (1) control,(2) 12 h fasting,(3) 24 h fasting,and (4) 48 h fasting groups.After animal models of fasting with different time courses were established,the heart,thoracic aorta,soleus,and extensor digitorum longus (EDL) were rapidly isolated from rats under anesthesia and then fixed into the respective organ perfusion system that was connected to physiological recorders and data acquisition system.Langendorff heart perfusion was used to detect some parameters of cardiac function,including left ventricular systolic pressure (LVSP),left ventricular end-diastolic pressure (LVEDP),and maximum ascending and declining rate of left ventricular pressure (± dP/dt max).Then,the heart was suffered from 30 min ischemia followed by 60 min reperfusion,the changes in above cardiac function parameters were recorded again.The contraction response to potassium chloride or phenylephrine and endothelium-dependent relation responses to acetylcholine (ACh) of aortic rings were measured with organ bath.The changes of twitch tension and titanic tension of soleus and EDL were examined to reflect contractive function of skeletal muscle.Results The + dP/dt max was significantly reduced in 24 h and 48 h fasting groups compared to the control (P ＜ 0.01).After ischemia-reperfusion,the heart re-beat time was significantly longer [(24.3 ± 7.0)min vs (14.0 ± 2.9)min,P ＜0.05],both mLVDP and mLVP were significantly higher in 48 h and 24 h fasting groups than control group (P ＜ 0.05),and those changes did not normalized until re-perfusion 60 min.The 48 h fasting increased significantly endothelium-dependent relaxation of aortic rings response to low concentration acetylcholine (P ＜ 0.05),but did not affect the maximum relaxation of aortic rings response to high concentration acetylcholine and contraction of aortic rings responses to potassium chloride and phenylephrine.The 48 h fasting also enhanced significantly titanic tension of soleus compared to the control [(7.01 ± 1.22) N/cm2 vs (5.5 ± 0.75) N/cm2 for48 h-Fasting vs Control,P ＜0.05].Conclusions The 24 ～48 h fasting may induce inhibition of cardiac function,while fasting 48 h can increase the sensitivity of aortic rings responses to ACh and contraction function of soleus in healthy and male rats.

ObjectiveTo investigate the preventive strategy of early post-operative complications of closed spinal dysraphisms in children.MethodsOne hundred and nine children with closed spinal dysraphisms underwent surgery from January 2004 to December 2008,were enrolled in this study.After dural closure,the wound was washed completely with saline to clear the tissue debris to prevent postoperative infection. We routinely made a horizontal incision to prevent postoperative infection. Results After operation,all patients were recovered and were discharged from hospitalNo post-operative complications,including cerebrospinal fluid leakage, wound infection and flap necrosis, were encountered. ConclusionThe sophisticated microsurgical technique and the delicate surgical manipulation is the prerequisite for the prevention of early operational complication of closed spinal dysraphisms in children.

Aim To investigate the effects of lentivirus mediated nerve growth factor ( NGF) gene silencing on pheochromocytoma cells ( PC12 ) and the possible mechanisms .Methods The NGF shRNA expression vector was constructed .PC12 cells were randomly divi-ded into five groups (n=3 each) as follows: negative control group ( NC ) , control lentivirus group ( LV CON) , lentivirus NGF shRNA1 group ( LV shNGF1 ) , lentivirus NGF shRNA2 group(LV shNGF2), lentivir-us NGF shRNA3 group(LV shNGF3).The cells in NC group were cultured in DMEM/HG and polybrene me-dium, while others were cultured in DMEM/HG, poly-brene and corresponding lentivirus medium .After the treatment, the infection efficiency was determined by fluorescent microscope .Relative expression of NGF , extracellular signal-regulated kinase ( ERK1/2 ) and p-ERK1/2 were assessed by Western blot .The expres-sion of NGF mRNA was analyzed by quantitative re-verse transcription polymerase chain reaction ( qRT-PCR) .The differentiation degree was valued according to the length of neuritis and max diameter of cells .The cell viability was detected by CCK-8.Results The in-fection efficiency in PC12 cells reached over 90%. Compared with NC group , the relative expression of NGF mRNA and NGF protein was significantly down-regulated ( P<0.05 ) .There was no difference in the expression of ERK1/2 protein and cell viability .The expression of p-ERK1/2 protein was markedly down-regulated in LV shNGF3 group ( P<0.01 ) .The cells morphology was changed , and the length of neuritis and max diameter of cells were strained in LV shNGF 3 group than those in NC group ( P<0.01 ) .Conclusion Lentivirus-mediated NGF gene silencing inhibits the differentiation of PC12 cells through suppressing the activation of ERK1/2.

Objective To investigate the effect of intrathecal morphine preconditioning on the expression of nerve growth factor (NGF) in the dorsal root ganglia (DRG) in a rat model of myocardial ischemia-reperfusion (I/R).Methods Thirty healthy adult male Sprague-Dawley rats in which intrathecal catheters were successfully placed without complications,weighing 250-350 g,were randomly divided into 5 groups (n =6 each) using a random number table:sham operation group (S group),I/R group,intrathecal morphine preconditioning group (ITMP group),μ receptor antagonist CTOP + intrathecal morphine preconditioning group (CTOP + ITMP group),and CTOP control group (CTOP group).Myocardial ischemia was induced by 30 min of occlusion of the anterior descending branch of the left coronary artery followed by 120 min of reperfusion in all the groups except S group.Intrathecal morphine preconditioning was produced by 3 cycles of 5 min intrathecal injection of morphine 3 μg/kg (10 μl) at 5 min intervals within 30 min before ischemia in ITMP group.In CTOP+ITMP and CTOP groups,1 μg/μ1 CTOP 10 μl was injected intrathecally at 10 min before morphine preconditioning and 40 min before ischemia,respectively.At 120 min of reperfusion,the rats were sacrificed,and myocardial specimens were obtained for determination of myocardial infarct size,and DRGs were removed for determination of the expression of NGF by using immunohistochemistry and Western blot.Results Compared with S group,the myocardial infarct size was significantly increased,and the expression of NGF in DRGs was significantly up-regulated in I/R group (P＜0.05).Compared with I/R group,the myocardial infarct size was significantly decreased,and the expression of NGF in DRGs was significantly down-regulated in ITMP group (P＜ 0.05),and no significant change was found in the parameters mentioned above in CTOP group (P＞0.05).Compared with ITMP group,the myocardial infarct size was significantly increased,and the expression of NGF in DRGs was significantly up-regulated in CTOP+ITMP and CTOP groups (P＜0.05).Conclusion The mechanism by which intrathecal morphine preconditioning reduces myocardial I/R injury is related to activation of spinal μ receptors,inhibition of NGF expression in DRGs,and reduction of responses to noxious stimulation in the rats.

Objective To investigate the effect of intrathecal morphine preconditioning (ITMP) on the excitability of substantia gelatinosa (SG) neurons in the dorsal horn of the spinal cord in a rat model of myocardial ischemia-reperfusion (I/R).Methods Thirty-six adult male Sprague-Dawley rats,weighing 200-300 g,in which intrathecal catheters were successfully placed without complications,were randomly divided into 3 groups (n =12 each) using a random number table:sham operation group (group S),group I/R,and group ITMP.Myocardial I/R injury was produced by occlusion of the left anterior descending branch of the coronary artery for 30 min followed by 120 min reperfusion.In group ITMP,the rats received intrathecal morphine 3 μg/kg (10 μl) by three cycles of 5 min infusions interspersed with 5 min infusion-free periods starting from 30 min before ischemia,and the equal volume of normal saline was given instead of morphine in group I/R.At 10 min of reperfusion,6 rats randomly selected in each group were sacrificed,and the T2-6 segments of the spinal cords were acutely isolated to prepare spinal cord slices.The resting potential,threshold of action potential (APT),peak of action potential (APP) and action potential duration in SG neurons in the dorsal horn of spinal cord slices were determined using the whole-cell patch-clamp technique,and the number of action potentials evoked by currents of 40,60,80 and 100 pA was recorded.At 120 min of reperfusion,6 rats randomly selected in each group were sacrificed,and myocardial specimens were obtained for determination of myocardial infarct size (IS) and area at risk (AAR),and IS/AAR ratio was calculated.The expression of c-fos in the T2-5 dorsal horns of the spinal cords was detected by Western blot.Results Compared with group S,the IS/AAR ratio was significantly increased,the expression of c-fos was up-regulated,the number of action potentials in SG neurons in dorsal horns of spinal cord was increased,APT was decreased,and APP was increased in group I/R (P＜0.05).Compared with group I/R,the IS/AAR ratio was significantly decreased,the expression of c-fos was down-regulated,the number of action potentials in SG neurons in dorsal horns of spinal cord was decreased,APT was increased,and APP was decreased in group ITMP (P＜0.05).Conclusion The mechanism by which ITMP attenuates myocardial I/R injury is related to decrease in the excitability of SG neurons in the dorsal horn of the spinal cord and reduction of responses to nociceptive stimuli in rats.

Acanthoma ; pathology ; Humans ; Keratosis ; pathology ; Skin Neoplasms ; pathology