Objective To investigate the risk factors of early allograft dysfunction (EAD) following C-Ⅱ donation after cardiac death (DCD) liver transplantation. Methods The data of 46 donors and recipients of C-ⅡDCD liver transplantation between March 2012 and August 2015 were retrospectively analyzed. The baseline data such as democracy, death cause, donor warm ischemic time (DWIT) and cold ischemic time (CIT) in EAD group and the non-EAD group (control group) was compared, and whether these factors were risk factors of EAD was investigated by univariate and multivariate analyses. Statistical cut-off values for significant factors of the unfavorable analysis were defined by receiver operating characteristics (ROC) analysis. The 6-month and 1-year graft survival rate were compared. Results The EAD group had a longer DWIT compared with the group [(17.6±4.7) and (12.7±6.2) minutes, P=0.009]; meanwhile, the EAD group had a longer CIT compared with the control group [(13.7±4.7) and (11.0±3.5) hours, P=0.020]. The other factors in both groups showed no statistical significance (P>0.05). The ROC curve revealed the cut-off values of DWIT and CIT were 17.50 minutes [area under the curve (AUC)=0.713, P=0.020] and 9.85 hours (AUC=0.723, P=0.015), respectively. The multivariate logistic regression analysis showed the DWIT [odds ratios (OR)=1.340, 95% confidence interval (CI)(1.042, 1.654), P=0.008] and CIT [OR=1.396, 95% CI (1.075, 1.698), P=0.015] were all independent risk factors of EAD. The 6-month and 1-year graft survival rate of the EAD group and the control group was 85.7% vs. 92.3% (P=0.607) and 71.4% vs. 84.6% (P=0.587), respectively. Conclusions EAD may occured in C-Ⅱ donors with DWIT≥17.50 minutes or CIT≥9.85 hours in DCD liver transplantation. The livers can be used as a resource for clinical use and also have a good outcome.
Objective To investigate the long-term clinical results of treatment of adult unicameral bone cyst with cancellous allograft. Methods From 1993 to 1998, 15 patients with unicameral bone cyst were treated by allograft with lyophilized cancellous bone. Among 15 patients, there were 5 males and 10 females, aging 19-41 years with an average of 27 years. The average follow-up time was 7.5 years (6-11 years). The X-ray films were taken and the CT scanning were carried out. Results The X-ray films showed that the allograft particles became vague 2-3 months after operation, that the allograft particles fused and began to form new bone and the bone density increased 5 months after operation, and that new bone formation completed after 7 months of operation. At the end of follow-up, remodelling in new bone occurred. Reoccurrence was not found in all patients. The symptom of pain disappeared or relieved obviously. Conclusion Allograft of lyophilized cancellous bone is an effective treatment for adult unicameral bone cysts.
【Abstract】 Objective To investigate the protective effect of early motion on articular cartilage after joint allograft by performing a controlled trial between different post-operation strategies after joint allograft in an animal model. Methods Twenty hemi-knee joints were harvested from 10 6-month-old New Zealand white rabbits (male or female, weighing 2.5-3.0 kg); 10 hemi-knee joints by deep frozen treatment (donors) were transplanted to unilateral knee joints (recipients) of 10 6-month-old Chinchilla rabbits (male or female, weighing 2.5-3.0 kg), which were divided into early motion group (n=5) and sustained fixation group (n=5); and 10 hemi-knee joints were used as blank control (n=5) and frozen control (n=5). The articular cartilage of allogenic joints was detected by X-ray film, gross, and histology at 6 weeks after operation. Results Gross observation: no obvious limitation of joint movements was observed in early motion group, but obvious limitation in sustained fixation group. X-ray films: the bone ends between donor and recipient healed well with good paraposition and alignment on the operation day and 2 weeks after operation; at 6 weeks, angulation deformity was observed in early motion group of 3 rabbits, and paraposition and alignment were satisfactory in sustained fixation group. Histological observation: HE staining showed that the chondrocytes had normal quantity and morphology with few nuclear fragmentation and karyolysis in early motion group, but the quantity of chondrocytes sharply decreased with dissolved nuclei and numerous fibrous tissues in the cartilage matrix in sustained fixation group. The cell survival rate of the early motion group (49.66% ± 2.15%) was significantly higher than that of the sustained fixation group (20.68% ± 1.24%) (P lt; 0.05). Scanning electron microscopy observation: nuclear membrane was intact with chromatin condensation and edema of mitochondria and rough surfaced endoplasmic reticulum in early motion group, and that the membrane of chondrocyte vanished with blurring border between chondrocyte and matrix, rupture of nuclear membrane and the disappearance of chromatin and organelles could be found in sustained fixation group. Conclusion Early motion has protective effect on articular cartilage after joint allograft, but cannot completely prevent degeneration of the allogenic articular cartilage.
Objective To investigate the preparing procedures for the larger chemically acellular nerve allografts (CANA) and to establish an evaluation criteria and the preparation method.Methods The sciatic nerves ofthe dogs were exposed by a muscle-splitting incision and were isolated free of the underlying fascia. The 50-mm-long segments of the nerve were made. The proximal and distal ends of the nerve segments were labelled and stabilized by pinning the ends to a thin plastic support, and then they were treated according to the following decellularization processes: The nerve segments were rinsed with the distilled water for 9h, transferred in a 3% Triton-100 solution for 12 h, soaked in 7% sodium deoxycholate for 12 h, and washed in the distilled water for 6 h. All the decellularization steps were performed at room temperature. The nerve segments were divided into 5 subgroups. In Group Ⅰ, Group Ⅱ and Group Ⅲ, the nerve segments were decellularized for 2, 3 and 4 times, respectively. In Group Ⅳ and Group Ⅴ, the two ends of the nerve segments were ligated with a silk line and were decellularized for 2 and 3 times, respectively. Each nerve segment was subdivided into 5 portions from the proximal end to the distal end. The degrees of decellularization, activity of Laminin, degrees of demyelination, and integrity of the nerve fiber tube were observed under microscope and were assessed by a scoring system. Results In all the groups the activity of Laminin was present and the degrees of decellularization were complete. As for the demyelination of the nerve segments, the myelin sheath in Groups Ⅰ, Ⅱ and Ⅲ was partially preserved, but it completely disappeared in Groups Ⅳ and Ⅴ. The structure of the nerve fiber tube in Groups Ⅰ and Ⅳ was almost normal. The total score for the degrees of decellularization, demyelination, and structural integrity was the lowest in Group Ⅳ but the quality was the best. Conclusion The degrees of demyelination are not parallel to the times of decellularization processes. In the quality control of CANA, we should consider the following 4 factors: activity of Laminin, degrees of decellularization, demyelination, and structural integrity. For the larger CANA,ligation of the two ends of the nerve segments during the decellularization procedure may be a better method of ensuring the quality of the decellularization.
Abstract: Objective To investigate the influence of cryopreservation on cellular viability of latepregnancy fetal valved allografts in human. Methods The fetal valved allografts with gestational ages ranged from 24 to 40 weeks were sterilely procured within 6 hours after brain death. Each sample was bisected into control group and experiment group. The cellular viability of control group was directly tested and that of experiment group was examined after being storaged in liquid nitrogen for a week through a programmed frozen procedure. The light microscopy, tissue culture and Methylthiazol tetrazolium assay (MTT assay) were used to determine the cellular viability. Results Twelve latepregnancy fetal valved aortic allografts were procured. Light microscopy showed the integrity of the basic structure of the thawed aorta, the normal structure of the collagen and elastic fibers, with part of vascular endothelium lost. There were lots of cells deriving from both groups,but the cellular growing rate of the experiment group was relatively slower. At 490 nm, MTT assay valve of control group was 0.442±0.046, and that of experiment group was 0.424±0.041. The difference between two groups failed to statistically significance(t=1.617, P=0.328). Conclusion There were viable cells in latepregnancy fetal valved allografts after cryopreservation.
To study the expression of CTLA4Ig gene in diabetic rat and the effect of CTLA4Ig on longterm survival of the pancreatic islet. The rat pancreatic islet cell and muscle cell transfected with the cDNA for CTLA4Ig packaged with lipofectin vector. We examined the expression level of CTLA4Ig gene, T lymphocyte reaction and observed the expression of CTLA4Ig cDNA in diabetic rat and the action of CTLA4Ig in longterm survival of pancreatic islet transplanted and the transplanted rats. Results: The T lymphocyte reaction from peripheral intravenous blood at seventh day after surgery, the difference between two group was significant (Ｐ＜0.05). Only 2 out of 10 recipients of the experiment group (A) at the seventh day after pancreatic islet allograft had any detectable levels of CTLA4Ig, their concentration was 14 ng/ml, and 31 ng/ml. The average time of maintaining blood glycose in normal levels of the group A after pancreatic islets graft, 14.8±12.3 days, was significantly longer than 3.6±5.1 days of the control group (B) (Ｐ＜0.05). The average survival time of the group A, 24.0±10.8 days (the longest time was 45 days), was significantly longer than 11.8±4.8 days (the longest time was 21 days) of the group B (Ｐ＜0.01). Conclusions: The muclse cells and pancreatic islets of the recipient rat was transfected with CTLA4IgcDNA packaged with lipofectin, and CTLA4IgcDNA was expressed in recipient tissue, its expressed product CTLA4Ig make pancreatic islets transplanted and recipient rat survive longer significantly.
Objective To investigate the clinical effects of repairing massive bone defects in limbs by using vascularized free fibular autograft compoundingmassive bone allografts. Methods From January 2001 to December 2003, large bone defects in 19 patients (11 men and 8 women, aging from 6 to 35 years) were repaired by vascularized free fibular transplant with a monitoringflap compounding massive deep frozen bone allografts. The length of bone defects were 12 to 25 cm (16.6 cm on average), of vascularized free fibular 15 to 28 cm (18.3 cm on average), and of massive bone allografts 11 to 24 cm (16.1 cm on average). Thelocation of massive bone defects were humerus in 1 case, femur in 9 cases and tibia in 9 cases. Results After followup of 5 to 36 onths (18.2 months on average), wounds of donor and recipient sites were healed at Ⅰstage, monitoringflaps were alive, no obvious eject reaction of massive bone allografts was observed and no complications occurred in donor limbs. The radiographic evidence showed union in 15 patients 3 months and 3 patients 8 months after operation. One case of malignant synovioma of left lower femur recurred and amputation was performed 2.5 months after surgery. Internal fixation was removed in 5 patients, and complete bone unions werefound 1 year postoperatively. No massive bone allografts was absorbed or collapsed. Conclusion With strict indication, vascularized free fibular autograft compounding massive bone allografts, as an excellent method of repairing massive bone defects in limbs, can not only accelerate bone union but also activate and changer the final results of massive bone allografts from failure.
ObjectiveTo explore the effect of hepatic outflow reconstruction with allograft vascular in ex-vivo liver resection and autologous liver transplantation.MethodThe clinical data of a patient with end-stage hepatic alveolar echinococcosis admitted to the Organ Transplantation Center of Sichuan Provincial People’s Hospital in August 2019 who underwent the ex-vivo liver resection and autologous liver transplantation combined with hepatic vein reconstruction with allograft vascular were analyzed retrospectively.ResultsThe patient, a 44-year-old female, was admitted to Sichuan Provincial People’s Hospital for “pain in the right abdomen accompanied by skin and sclera yellow staining for 6+ months and aggravated for 20+ d”. When the patient was admitted, the general condition was poor, such as hyperbilirubin and hypoproteinemia. The body mass was 45 kg and the standard liver volume was 852 mL. The hydatid lesions corroded the first and second hilum of the liver, the right hepatic vein and the posterior inferior vena cava. It was difficult to reconstruct the outflow tract of the hepatic vein in vivo, and it was extremely difficult to completely remove the hydatid lesions in vivo. After admission, the patient was generally in a good condition after the PTCD treatment, then after discussion and rigorous evaluation, the ex-vivo hepatectomy combined with autologous liver transplantation was required. The operative time was 15 h and the intraoperative blood loss was approximately 2 000 mL. After the operation, the routine treatment was performed, the antiviral treatment was continued, the international standardized ratio value was monitored at 1.5–2.5, and the anti-immune rejection drugs were not needed. The patient was transferred to the general ward on the 4th day after the operation, and there were no bile leakage, bleeding, infection and other complications. the result of postoperative pathological diagnosis was the alveolar echinococcosis. The re-examination of enhanced CT on 1 week after the operation suggested that the hepatic outflow tract of allograft vascular reconstruction was unobstructed, no stenosis and no thrombosis occurred. The patient was following-up at present.ConclusionsIn treatment of end-stage hepatic alveolar echinococcosis by autologous liver transplantation, reconstruction of hepatic outflow should be individualized. Allograft venous vessels could be used as ideal materials due to their advantages of matched tube diameter and length, no anti-rejection, and low risk of infection.
Objective To investigate the rat model of cardiac allograft vasculopathy after heart transplantation in rat abdominal cavity. Methods Forty Wistar rats and 40SDrats were divided into control group and experiment group randomly pair-matching. Rat model ofheterotopic heart transplantation was developed. Low doseCyclosporine A were injected into the abdominal cavity in experiment group, while the control group had not received the Cyclosporine A. Transplant hearts were harvested at two weeks and four weeks post-operatively and changes of coronary artery were observed by light microscope. Results There were no alteration of tunica intima of coronary artery in control group at two weeks and four weeks post-transplantation. Tunica intima of coronary artery increased in thickness at two weeks post-transplantation in experiment group and concentric circular change occurred at four weeks post-transplantation. Lumen of coronary artery constricted transparent and cardiac allograft vasculopathy occurred. Conclusion This animal model is reliable of cardiac allograft vasculopathy.
ObjectiveTo compare the half-year clinical efficacy of three different surgical root coverage methods including lateral sliding flap (LSF), subepithelial connective tissue graft (CTG) and acellular dermal matrix allograft (ADM). MethodsEighteen patients (24 teeth) with gingival recession treated in our hospital between December 2012 and July 2015 were selected and divided into three groups according to a certain sequence with 8 teeth in each group. The three groups of teeth were treated with LSF, CTG and ADM respectively. Gingival recession, probing depth and keratinized gingival width at both baseline and 6 months after surgery of all patients were recorded for inner- and inter-group comparison. ResultsAll three methods proved to be effective within 6 months with an awerage of 2.8-4.0 mm in decrease extent of gingival recession (P<0.01). LSF did not significantly change the probing depth (P>0.05) as the other two did (P<0.01). The differences among three surgical methods compared before and after surgery were all significant (P<0.05). ConclusionLSF, CTG and ADM are all effective surgical means for root coverage. Within 6 months, CTG presents better effects than ADM, and ADM better than LSF.