The Airflow Characteristics in a 3D Airway Model Based on CT Images / 医用生物力学

Objective To discuss the mass transfer of low temperature gas in the lung bronchus, so as to provide a theoretical basis for the implementation of hypothermic ventilation cooling non-heart-beating donor (NHBD) lung program. Methods A real airway model was reconstructed based on human lung CT images, and the computational fluid dynamics (CFD) method was used to investigate the airflow characteristics inside the airway during reciprocating ventilation. The effect of ventilation frequency (0.5, 0.25, 0.125 Hz) on bronchial flow was also studied. Results The flow in the airway showed complex three-dimensional （3D） flow characteristics during reciprocating ventilation. The flow in different areas of the airway was different during inhaling and exhaling; the irregular bronchial geometry had an important effect on its internal flow; when the ventilation frequency decreased from 0.5 Hz to 0.125 Hz, the thickness of flow boundary layer would increase, and the mainstream velocity in different areas of the airway was enhanced to different degrees. Conclusions The real airway model based on CT 3D reconstruction was more accurate than the ideal circularity tube model in showing the bronchial flow. The research findings have an important guiding significance to optimize the hypothermic ventilation cooling NHBD lung technique.

Protective effects of ulinastatin on ischemia-reperfusion injury during rat non-heart beating donor lung transplantation / 中华器官移植杂志

Objective To investigate the protective effects of ulinastatin (UTI) on ischemiareperfusion injury of donor lungs,and the possible mechanism.Method Forty male SD rats were randomly divided into two groups:group C as control group and group U as UTI group.In group C donor lungs were antegradely flushed with 20 ml of cold (4 C) low potassium dextron (LPD) solution and 5 ml retrogradely.Meanwhile,in group U,UTI (500 000 U/L) was added in LPD solution and the same doses were used.According to the time after initiation of reperfusion,each group was divided into two subgroups:30 min (subgroup A) and 1 h (subgroup B).Arterial blood samples were collected for blood gas analysis.Lung samples were obtained at the end of reperfusion (30 min or 1 h).Microscopic examination of the donor lungs was conducted.Besides,the pulmonary water index (W/D),tissue malondialdehyde (MDA) and superoxide dismutase (SOD) content,and mRNA expression of tumor necrosis factor (TNF-a),intercellular adhesion molecule 1 (ICAM-1) and interleukin 10 (IL-10) were also measured.Results (1) One h after reperfusion,oxygenation index in group U was higher than that in group C (P =0.025) ; (2) The levels of W/D in subgroup A and subgroup B of group U were decreased as compared with group C (P =0.005 and P =0.006) ; (3)The microscopic changes of donor lung tissues in group U were lessen than in group C; (4) In subgroup A of group U,MDA content was decreased (P=0.039),and SOD content was increased (P=0.035),and similar results could be observed in subgroup B of group U (P =0.006 and P =0.030 respectively); (5) As compared with group C,the mRNA expression of TNF-α in group U was decreased at the time of 30 min after reperfusion (P =0.000),but no significant change was found at the time of 1 h (P =0.139).The mRNA expression of ICAM-1 was not decreased evidently at the time of 30 min (P=0.062),but significantly decreased at the time of 1 h (P=0.001).The mRNA expression of IL-10 was increased in subgroups A and B (P =0.004 and P =0.000 respectively).Conclusion This study demonstrated that UTI had protective effects of reducing ischemia-reperfusion injury on the donor lungs after lung transplantation in rat non-heart beating donor models.

Evaluation of liver grafts with warm ischemia and with different cold preservation time in liver transplantation / 中华消化外科杂志

Objective To evaluate the efficacy of liver grafts with warm ischemia and with different cold preservation time in liver transplantation.Methods The clinical data of 154 patients who received liver transplantation at the First Affiliated Hospital of Sun Yat-sen University from January 2006 to December 2007 were retrospectively analyzed.The warm ischemia time of the liver grafts obtained from the non-heart-beating donors was within 10 minutes.According to cold perservation time of the liver grafts,patients were divided into 3 groups:the cold preservation time of the liver grafts was within 8 hours,8-12 hours and above 12 hours in group I(n=58),group Ⅱ(n=62)and group Ⅲ(n=34),respectively.The peak level of alanine aminotransferase(ALT),primary graft dysfunction(PGD)after liver transplantation,acute rejection response,biliary complications,vessel complications,perioperative infections and the survival of liver grafts and recipients among the 3 groups were analyzed via chi-square test,t test and variance analysis.Results No PGD was detected in the 3 groups after liver transplantation.All patients were followed up for 8-32 months.The peak level of ALT,incidence of infection and biliary complication,survival of liver grafts and recipients were(482±357)U/L,12%(7/58),12%(7/58),86%(50/58)and 88%(51/58)in group Ⅰ,and were(1274±608)U/L,29%(10/34),26%(9/34),68%(23/34)and 71%(24/34)in group Ⅲ,with significant difference between the 2 groups(t=5.23,X~2=4.28,6.77,4.51,4.28,P<0.05).The peak level of ALT in group Ⅱ was(953±424)U/L,which was significant higher than(482±357)U/L in group Ⅰ(t=4.76,P＜0.05).Conclusions Liver grafts with a warm ischemia time shorter than 10 minutes could tolerate the injury caused by cold preservation with the maximum time of 12 hours.The incidences of biliary complications and postoperative infections are significantly increased and the survivals of liver grafts and recipients are decreased when the cold preservation time exceeds 12 hours.

Effects of HGF in Celsior solution on levels of INF-γ. IL-4 and IL-10 in a rat liver transplantation model / 中华肝胆外科杂志

Objective To explore the impact of recombinant human hepatocyte growth factor (rhHGF) in Celsior (CS) solution on the expression of INF-γ, IL-4 and IL-10 in a rat liver transplan-tation model. Methods After flushed with CS solution with addition of rhHGF (experimental group) or saline (control group), NHBD livers were stored at 4℃; for 16 h.then they were transplanted using the two-cuff technique with arterial reconstruction. The serum levels of INF-γ, IL-4 and IL-10 at lh after reperfusion were detected using ELISA. The INF-γ, IL-4 and IL-10 mRNA in the corresponding liver tissue were determined by RT-PCR. The 7-day survival rate was calculated and the histopatho-logical examination results were analyzed by hematoxylin and eosin staining. Results Compared with the control group, the experimental group showed lower INF-γ level and higher IL-4 and IL-10 levels in serum at 1 h after reperfusion (P<0. 05). The level of INF-γ mRNA in liver tissue was significant decreased at 1 h after reperfusion (P<0. 05) , and the level of IL-4 and IL-10 mRNA was significantly increased in the experimental group (P<0. 05). In experimental group, recipients got a better survival rate and histopathological examination showed a well-preserved hepatic architecture without hepatocyte necrosis, milder sinusoidal and portal congestion. Conclusion Adding exogenous rhHGF in CS solu-tion can protect NHBD livers from ischemia-reperfusion injury and prolong the survival in rats, which might be due to down-regulation of TNF-γ and up-regulation of IL-4 and IL-10.

Anesthesia for liver transplantation from a maastricht category 4 non-heart-beating donor: A case report / 대한마취과학회지

Great improvements in patient selection, surgical techniques, perioperative care, and immunosuppression have been made for the optimization of liver transplantation. To increase the number of organs available for liver transplantation, transplant centers have used marginal donors, split livers, living donors, or non-heart-beating donors (NHBDs). Despite recent enthusiasm for NHBDs in liver transplantation, warm ischemic injury to recovered organs has been an obstacle for the wide acceptance of NHBD. In the present case, we have conducted a liver transplantation from a Maastricht Category 4 NHBD. Warm ischemic time was 20 minutes and cold ischemic time was 5 hour 43 minutes. Consequently, the liver was successfully transplanted into the recipient.

The Use of Non-Heart Beating Donors to Expand the Donor Pool / 대한이식학회지

There is a persistent shortage of allografts available for transplantation, so we envisioned using non-heart beating donation to expand the donor pool. Non-heart beating donors (NHBD) were categorized using four definitions. Controlled donors, consisting of categories III and IV, are the most suitable for NHBD. Delayed graft function is associated with the use of kidneys from such donors, but had no difference on graft survival in the long-term results compared with heart beating donors. The proportion of NHBD of deceased donors differs considerably among countries, but national programs in many nations have now been initiated to increase the rate of non-heart beating donation. In most cases, the organs from NHBD are not available for transplantation in Korea because of legal restrictions. The use of controlled NHBD is encouraged to expand available allografts in Korea, due to the shortage of such allografts

Induction of apoptosis in non-heart-beating donor livers by peroxynitrite / 中华肝胆外科杂志

Objective To investigate the role of peroxynitrite in induction of apoptosis in non-heart-beating donor (NHBD) liver grafts. Methods Rat livers were harvested and perfused via the portal vein with 60 ml of cold 4℃ HTK solution from heart beating donors (HB) or 60 min after car-diac arrest from NHBD. A third group consisted of NHBD livers which were perfused with HTK con-taining 7500 IU of superoxide dismutase (SOD) (n=6, resp.). After cold storage for 24 h, the func-tional integrity of the liver was evaluated by isolated reperfusion with Krebs-Henseleit Buffer in a re-circulating system at 37℃ for 45 min. Results Compared to the NHBD livers, HB and SOD groups were characterized by a significantly reduced hepatic enzyme leakage(P<0. 05) as well as decreased portal venous pressure(P<0.01). Bile production upon reperfusion was increased significantly (P< 0.01). Prominent decreasing nitrite oxide(NO) (P<0.05), nitrotyrosine(P<0.01, P<0.05) and apoptosis(P<0.05) in effluent perfusate was encountered. Slight nitrotyrosine staining was concen-trated on the endothelial cells. Conclusion Peroxynitrite may play a fundamental role in the apoptosis caused by enhanced oxygen free radical production in NHBD livers.

Intrahepatic diffuse biliary stricture after orthotopic liver transplantation / 中华普通外科杂志

Objective To evaluate the treatment and prevention measure for intrahepatie diffuse biliary stricture after orthotopic liver transplantation. Methods The clinical data of 21 patients with intrahepatic diffuse biliary stricture after orthotopic liver transplantation from January 2002 to December 2007 in Beijing Center for Organ Transplantation were retrospectively analyzed. A prospective clinic study was launched in order to prevent microthrombosis in the microcireulation of bile duct during warm and cold iscbemia in non-heart-beating donor since January 2006. Urokinase perfusion of the artery system was conducted during graft harvesting and reperfusion at the end of graft trimming to reduce the incidence of intrahepatic diffuse biliary stricture. Results Among 21 patients with intrahepatie diffuse and mixed type biliary stricture, 16 patients underwent liver retransplantation, and the other 5 patients died of primary graft failure while waiting for retransplantation. The incidence of intrahepatic diffuse biiiary stricture was 5.9% in non-urokinase perfusion group. On the contrary, the incidence rate of intrahepatie diffuse biliary stricture was 1.4% in urokinase perfusian group (x2 = 5.98, P < 0. 05). Conclusions Liver retransplantation is effective for refractory biliary stricture in liver transplant recipients. The incidence of intrahepatic diffuse biliary stricture is reduced in non-heart-beating donor by using urokinase perfusian.

Detection of Bronchial Function of NHBD Lung Following One-h Warm Ischemia by Organ Bath Model / 华中科技大学学报（医学）（英德文版）

laxant abilities of bronchial smooth muscles, and the epithe-lium-dependent adjustment both kept intact. Organ bath model could be a liable and scientific way to evaluate the bronchial function of NHBD lung.

Liver Transplantation Using Non-Heart Beating Donor: The First Korean Case Report / 대한이식학회지

A liver originating from Maastricht category 4 non-heart-beating donor (NHBD: cardiac death in a brain death donor) was procured and transplanted. Donor was 46 years old female. She was moved to the operation room after 3 times of cardiopulmonary resuscitation. Arrest occurred 15 minutes after stopping ventilation. After 5 min waiting time, the incision was performed. The interval between incision and initiation of donor perfusion was 5 minutes. Warm ischemic time, which is from the withdrawal of support to perfusion, was 25 minutes. Super-rapid technique was used for the donor procedure. The frozen biopsy of the liver was performed before transplantation and macrovesicular and microvesicular fatty change were less than 5% respectively. The cold ischemic time was 6 hours 22 minutes. Orthotopic liver transplantation was performed with the preservation of the recipient caval vein without venovenous bypass. The recipient was 56 years old female. She suffered from cryptogenic liver cirrhosis with refractory ascites. Postoperatively, the early graft function was good. At the post-operative 10th day, Serum total bilirubin was 1.4 mg/dL and aspartic acid transaminase and alanine aminotransferase was 26 IU/L and 20 IU/L respectively. Post operative 10th day liver biopsy was normal. She stayed at the intensive care unit for 6 days. Post-operatively, Tuberculosis (Tb) peritonitis (by the intra-operative omentum tissue culture) was diagnosed and the patient is under Tb medication. This experience suggests that careful donor selection, minimizing warm and cold ischemic time and utilization of histology provide acceptable results of liver transplantation from NHBD.

The Feasibility of Recovered Warm Ischemic Injury on Non-heart-beating Donor Liver by Hepatocyte Growth Factor in Animal Model / 대한이식학회지

PURPOSE: Liver transplantation is the therapy of choice for patients with acute and acute-on-chronic severe liver failure or hepatocellular carcinoma. But a suitable liver is not always available for transplantation due to limited donor numbers. To increase the number of available liver for transplantation, a non-heart-beating donor (NHBD) liver transplant program is started. In NHBD liver transplantation, warm ischemic injury of liver occurs. The duration of warm ischemia is thought to be the most important risk factor for postoperative complications such as primary nonfunction or severe hepatic dysfunction. Recent evidence indicates that hepatocyte growth factor (HGF) plays an important role as a cytoprotector against hepatic injury by anti-apoptotic effect and mitogen in liver regeneration. Therefore studies also were performed to examine whether HGF influenced the viability and regeneration of hepatocytes from rats, subjected to prolonged warm ischemic injury. METHODS: Male Sprague- Dawley rats were subjected to non-heart-beating death by cervical spine fracture. Rats left in room temperature directly after, 30-minutes, 1-hours before surgery and perfusion was performed for isolating hepatocyte. Among three groups, hepatocyte viability was compared by trypan blue stain. And isolated hepatocytes from 30-minutes warm ischemic group were cultured for 24-hours, which were treated with no HGF and addition of various doses (5 ng/mL, 10 ng/mL, 20 ng/ mL, 40 ng/mL, 100 ng/mL) of HGF. Anti-apoptosis and regeneration of hepatocyte were compared by LDH assay, MTS assay, western blot, and immunocyto-chemistry after a 24-hours culture. RESULTS: The results of hepatocyte viability along the prolonged warm ischemic groups in isolated hepatocytes decreased sequentially 74.8+/-12.6%, 45.0+/-5.4%, 37.8+/-10.4% along directly after, 30-minutes, 1-hours in trypan blue stain (P<0.01). And 24-hour-cultured hepatocytes from 30-minutes warm ischemic group were treated with HGF. The results of LDH assay, MTS assay did not have relation with HGF addition. But the results of western blot and immunocytochemistry shown that HGF doses dependent anti-apoptosis and regeneration of hepatocyte increased. That indicates HGF presumably inhibites apoptotic pathway by phosphorylation. And HGF also makes hepatocyte hypertrophy and albumin synthesis. CONCLUSION: HGF was a potent cytoprotector against hepatic injury by anti- apoptotic effect and mitogen of liver regeneration in NHBD liver animal model. HGF facilitates recovery of the liver from prolong warm ischemic injury. If the more clinical studies and large animal studies are performed, NHBD using liver transplantation will be available with more chances by HGF.

A Porcine Model for Non-heart Beating Donor Liver Transplantation / 대한이식학회지

PURPOSE: Non-heart-beating liver transplantation has been accepted as a substitute to overcome the donor shortage problem. However, prolonged warm ischemia during liver procurement remains a obstacle to widespread use of non- heart-beating transplantation. Therefore, experimental studies to ameliorate graft injuries have been conducted, but, their clinical applications are not satisfactory yet. The aim of this study is to test our experimental model as a pertinent non- heart-beating transplantation model. METHODS: We designed porcine non-heart-beating liver transplantation model by simultaneous liver procurement of donor and recipient. Cardiac death was induced by direct cardiac injection of potassium chloride. Perfusion of normothermic hypertonic saline started after 30-min (group A, N=5), 1-hour (group B, N=4). Orthotopic liver transplantation with perfused donor liver was performed and we compared the perioperative laboratory parameter, histologic findings and survival between two groups. RESULTS: Only one (11.1%) death occurred among the nine transplant pigs. 2-day survival rates of group A and B were 60%, 50%, respectively. Group A showed a relatively acceptable posttransplant laboratory findings including liver function test and normal-looking histologic feature at the time of reperfusion and 24 hours after reperfusion. Group B showed more deranged liver function test and ischemic liver cell morphology at 24 hours after reperfusion. CONCLUSION: Our results suggest that this porcine non-heart-beating transplantation model may be the safe and suitable method. This model will be useful in further study for testing the perfusate and drugs to ameliorate the warm ischemia-induced hepatic injury.