Objective To study the effect of Huaier granule on the recurrence and metastasis of hepatocellular carcinoma (HCC) and immune rejection in the postoperative patients with liver transplantation for HCC. Methods Twenty-eight patients of liver transplantation for HCC who had taken Huaier granule orally for more than 6 months from September 2001 to March 2007 in West China Hospital were included as treatment group, and other 56 patients of liver transplantation for HCC who didn’t take any Huaier granule in the same time were included as the control group according to the same stage of TNM, degree of tumor differentiation (Edmondson grading) respectively with the treatment group. The method of retrospective cohort study was used to compare the incidence of immune rejection and the 6-month, 1-year, and 2-year recurrence and metastasis of HCC, disease free survival rate, and survival rate between two groups after 2 years’ follow-up beginning from the date of surgery. Results The 6-month, 1-year, and 2-year tumor recurrence and metastasis incidences in treatment group were 14.3%, 32.1%, and 39.3% respectively, which were 23.2%, 32.1%, and 50.0% respectively in control group, and the 2-year tumor recurrence and metastasis incidence of the treatment group was lower than that of the control group. The 6-month, 1-year, and 2-year disease free survival rates in treatment group were 85.7%, 67.5%, and 60.0% respectively, which were 76.7%, 67.6%, and 49.3% respectively in control group, and the 2-year disease free survival rate of treatment group was higher than that of the control group. The 6-month, 1-year, 2-year survival rates in treatment group were 92.9%, 78.6%, and 67.9% respectively, which were 89.3%, 75.0%, and 62.5% respectively in control group. But the 2-year tumor recurrence and metastasis incidence (P=0.353), 2-year disease free survival curve (P=0.386), and 2-year survival curve (P=0.620) were not significantly different between two groups. The incidence of immune rejection was 14.29% in the treatment group and 16.07% in the control group, which was not significantly different between the two groups (P=0.831). Conclusions Huaier granule can increase the 2-year tumor-free survival rate and restrain the recurrence and metastasis of HCC, and does not increase the incidence of immune rejection. Huaier granule as a treatment of HCC in patients with liver transplantation is safe and effective.
Objective Tri ptol ide can suppress immunological rejection reaction. To investigate the effect of tri ptol ide on allogenic tendon transplantation in repairing tendon defect in chicken. Methods The defect model of the third toes tendon was establ ished in 64 healthy-cleaning male Leghorn chickens (4-month-old, weighing 1.9-2.3 kg), which underwent allogenic tendon transplantation for repairing and were divided into 2 groups randomly (n=32). Tri ptol ide feeding[100 μg/(kg·d)] was given for 3 weeks in the experimental group and normal feeding in the control group. General condition of the chickens was observed after operation. The transplanted tendons were harvested from 4 chickens in each group for gross observation at 1, 2, 3, and 4 weeks after operation; the histological observation was performed at 1 and 3 weeks, and transmission electron microscope observation at 2 and 4 weeks. The blood and tendon were harvested from another 8 chickens in each group for flow cytometry and biomechanical tests respectively at 3 and 6 weeks. Results All chickens survived to the experiment end. Gross observation: with time extending, hyperemia and edema around transplanted tendon were rel ieved. Rarefaction adhering zone was seen in experimental group, and pyknotic adhering zone in control group. Histological observation: inflammatory reaction in experimental group was sl ighter than that in control group at 1 and 3 weeks. Transmission electron microscope observation: at 2 and 4 weeks, fibroblasts had big cell nucleus, more euchromatin, and l ittle heterochromatin in experimental group; however, there were small amount of rough endocytoplasmic reticulums with gentle expanded capsular space in control group, which contained sparse content. Flow cytometry test: at 3 and 6 weeks, peri pheral blood contained less CD4+ and CD8+ T lymphocytes in experimental group than in control group, and the ratio of CD4+ to CD8+ T lymphocyte significantly decreased in experimental group when compared with control group (P lt; 0.05). Biomechanical examination: at 3and 6 weeks, the maximum tensile strength in experimental group was bigger than that in control group, and tensile adhesion power in experimental group was smaller than that in control group. There were significant differences in the indexes between 2 groups (P lt; 0.05). Conclusion Tri ptol ide can suppress immunological rejection reaction, strengthen tendon healing strength, and reduce tendon adhesion in allogenic tendon transplantation.
Objective To summarize the research progress of xenotransplantation.Methods Domestic and international publications about xenotransplantation were summarized and reviewed. Results Hyperacute xenograft rejection was a huge problem for xenotransplantation， but it could be alleviated if the organs or tissues of donor were genetically modified. So far the graft survival time differed greatly due to characteristics of different organ. Conclusions By reviewing the studies of relevant papers about xenotransplantation， a comprehensive understanding of research background and a suitable research direction of xenotransplantation can be supplied. The graft organs or tissues from genetically modified donors are expected to avoid or alleviate hyperacute xenograft rejection.
Objective To investigate the mechanism of hyperacute rejection (HAR) in pig to rhesus monkey vein xenograft. Methods Porcine femoral vein was transplanted into rhesus monkey. Deposits of IgM, IgG, C3 and C4 on the grafts were observed by immunoflurescence. Results Great deal of IgM, C3 and C4 were seen along the endothelium of donor vein, but IgG was not seen. ConclusionIn pig to monkey xenograft model, HAR is intiated by the binding of xenoreactive IgM to donor xenoantigens and followed by the activation of complement via the classical pathway.
OBJECTIVE:To investigate the index of the rejection of lJle retinal pigment epithelium(RPE)cells transplantation. METHOD:Allogenic RPE transplantation on rahbits by transcleral technique, the changes of interleukin-2 (IL-2) activity in peripheral blood and the effect of immunoinhibitor (methylprednisonlone)were detected. RESLILTS:In the group of simple transplantation,the IL-2 activity in peripheral blood begin to rise in the first day after operation. The peak value occured in the third day,and is still much higher than that of the control group in the 14th day,whereas in the group treated with immunoinhibitor ,there was no obvious difference in the first day after operatlon,in the third day,the IL-2 activity rises slightly,and returned to normal level in the 7th day. CONCLUSION: After RPE transplantation, the level of IL-2 activity in peripheral blood might serve as an important index to determining and detecting the rejective response. （Chin J Ocul Fundus Dis,1996,12: 239-241）
Insufficient supply of organ for allotransplantation made the study on finding new organ resources from animal progress. Pig is regarded as one of the optimal donor animals for human. The major obstacle in this field is hyperacute reaction (HAR), which is triggered after the xenogenic natural antibodies preexisting in recipient blood combine to the antigens on the surface of the endothelium and activate the complement system. alpha-Galactose residues (alpha-Gal) on the endothelial cell have been identified as the major xenoantigens. NJZ Pig has been closely breed since 1938, whose family history is clear. Tissue samples from heart, liver, kidney, pancreas, lung, small intestine, skin, spleen, thymus and lymph node were obtained and embedded in paraffin. The sections were performed the immunohistochemical staining with the sera from health volunteers (including all the blood types) as the primary antibodies as well as the biotin labeled bandeirae simplicifolia I isolectin B4 (BS I-B4), which has specific affinity to alpha-galactose. All the staining sections were compared with the tissues digested with alpha-galactosidase. There was no difference between the antigens recognized by sera of different blood types. alpha-Gal was still the major xenoantigen on the endothelial cells. There might exist non-alpha-Gal antigens on the distal convoluted tubules and collecting tubules of the kidney. There was no alpha-Gal distributing on the secreting part of pancreas, either the islet cells or the matrix cells, but surely on pancreatic duct and vessels. All the antigenity was destroyed after the enzyme digestion except that the small intestine gland still positive with the BS I-B4. alpha-Gal is the major xenogenic antigen in NJZ Pigs. There exist some unknown antigens on the distal convoluted tubules and collecting ducts of the kidney. The blood type of recipient is not the first affair to be considered in pig-to-human xenotransplantation. The specificity of BS I-B4 for the alpha-galactose needs more detail research.
Objective To investigate the immunological rejection after hepatocyte transplantation for acute liver failure （ALF） in mice．Methods The hepatocytes were isolated from pig，BALB/c and C57BL/6 mice livers were conducted and then transplanted into C57BL/6 mice．CCl4 was used to make ALF mice model．The experimental animals were randomly divided into three groups， including syngenic group，allogeneic group，and xenogenic group．The survival statuses of all the mice were recorded. The alteration of T lymphocyte subsets，immune globulin，and cytokine were determined．Results ①The survival ratio was 8/10，6/10， and 3/10 in the syngenic group， allogeneic group， and xenogenic group， respectively．The survival ratio in the syngenic group was significantly higher than that in the other two groups （P＜0.05）．②The CD4+ and CD8+ T cells of the peripheral blood in the syngenic group did not change significantly on week one after transplantation．The CD4+ T cells in the allogeneic group reached the peak on day 3 after hepatocyte transplantation （P＜0.05）， while CD8+ T cells did not change much in one week．The CD4+ and CD8+ T cells in the xenogenic group increased and reached the peak on day 3 after transplantation （P＜0.05）．③There were no significantly differences of IgM and IgG in the syngenic group among 0.5， 1， and 3 d after transplantation. IgM of the allogeneic group and xenogenic group reached the peak on day 1 （P＜0.05） and IgG reached the peak on day 3 （P＜0.05） after transplantation．④The concentrations of IFN-γ， TNF-ɑ， and IL-2 in the allogeneic group and xenogenic group were significantly higher than those in the syngenic group （P＜0.05）．The concentration of IL-6 of the xenogenic group was higher than that of the other two groups （P＜0.05）. Conclusions CD4+ and CD8+ T cells play an important role in immune response to both allogeneic and xenogenic hepatocyte transplantation， as well as induce humoral immune response early after hepatocyte transplantation．
Objective To establ ish the modified model of cervical heterotopic cardiac transplantation in rats for investigation of cardiac chronic rejection. Methods Forty healthy male Wistar rats, aged 10 weeks, weighing 250-300 g, were appl ied as the donor group, and forty healthy male SD rats, aged 10 weeks, weighing 300-350 g, served as the recipient group. The donors’ pulmonary artery was anastomosed to the reci pients’ right external jugular vein by non-suture cuff technique while the donors’ innominate artery was anastomosed to the recipients’ right common carotid artery by suture microvascular anastomosis. All recipients received cyclosporin to prevent acute allograft rejection. Results Forty consecutive successful transplantations were performed. Neither anastomosis leakage nor vessel obstruction occurred. The total operation time was 40-50 minutes. The time of cuff vascular anastomosis was 2-3 minutes and that of microvascular anastomosis was 9-12 minutes. All recipients survived for more than 30 days and all allografts were examined at 30 days after the transplantation. Pathological manifestations of allograft vessels were chronic rejection. Conclusion This modified model of cervical heterotopic cardiac transplantation is simple, practical and highly reproducible and is appl icable for investigation of chronic rejection in various organ transplantation studies.
Objective To evaluate the long-term function of the traumaticallydamaged joint after its repair with transplantation of a fresh or a frozen allogenic joint. Methods From March 1977 to September 1993, 13 patients (9 males, 4females; age, 17-55 years) with traumatically-damaged joints underwent transplantation of the fresh or the frozen allogenic joints. Five patients had 5 damagedmetacarpophalangeal joints, 6 patients had 9 damaged interphalangeal joints, and 2 patients had 2 damaged elbow joints. So, the traumatic damage involved 13 patients and 16 joints. All the metacarpophalangeal joints and the interphalangeal joints were injured by machines and the 2 elbow joints were injured by road accidents. The patients were randomly divided into 2 groups: Group A (n=7) andGroup B (n=6). The 7patients with 8 joints in Group A underwent transplantation of fresh allogenic joints; the 6 patients with 8 joints in Group B underwent transplantation of frozen allogenic joints. The allogenic joint transplants were performed in the period from immediately after the injuries to 6 months after the injuries. The motion ranges of the transplanted joints and the X-ray films were examined after operation, and the immunological examination was performed at 8 weeksafter operation. Results The time for synostosis was 5-8 months in Group A, but4-6 months in Group B. In Group A, at 2 years after operation the metacarpophalangeal flexion was 30-40° and the interphalangeal flexion was 20-30°; however,at 6 or 7 years after operation the interphalangeal flexion was only 10-20°. The patients undergoing the transplantation with fresh elbow joints had the elbowflexion of 60° and the elbow extension of 0°, and had the forearm pronation of 30°and the forearm supination of 30°. But in Group B, at 2 years after operation the metacarpophalangeal flexion was 6070° and the interphalangeal flexionwas 40-50°; at 6 or 7 years after operation the interphalangeal flexion was still 40-50°. However, the patients undergoing the transplantation with frozen elbow joints had the elbow flexion of 90° and the elbow extension of 0°, and hadthe forearm pronation of 45° and a forearm supination of 45°. The joint motion ranges, the Xray findings, and the immunological results in the patients undergoing the transplantation of the frozen allogenic joints were significantly better than those in the patients undergoing the transplantation of fresh allogenicjoints. There was a significant difference in the immunological examination between Group A and Group B (IL2, 21.64±3.99；CD4/CD8,3.88±0.82 vs.IL-2,16.63±3.11；CD4/CD8, 2.53±0.23, P＜0.01). Conclusion Repairing the traumatically-damaged joints with frozen allogenic joints is a better method of regaining the contour, movement, and complex motion of the hands.
Objective To investigate the effect of human placental-derived mesenchymal stem cells (PMSCs) on immunological rejection in mouse allogeneic skin transplantation. Methods The placenta fetal tissues from voluntary donors were used to isolate and culture the PMSCs, and the 3rd passage PMSCs were used in the experiment. Thirty Vr ∶ CD1 (ICR) mice at age of 1-2 days were used as skin donors for allogeneic skin transplantation. Thirty C57BL/6 mice at age of 6-8 weeks as recipients were made back skin defect of 12 mm in diameter and were randomly divided into 3 groups (n=10): group A, autograft; group B, allogeneic graft + PBS tail vein injection; and group C, allogeneic graft + human PMSCs (1 × 105 cells/mouse) tail vein injection. The flap survival was observed. At 7 days after skin transplantation, blood leukocyte counting, abdominal fluid macrophage activation, and the expression levels of interleukin 4 (IL-4), interleukin 17 (IL-17), and interferon γ (INF-γ) in blood and spleen were detected by ELISA and RT-PCR, respectively. Results The flap survival time was significantly longer in group A [(58.33 ± 4.04) days] than in groups B and C [(3.80 ± 0.92) days and (6.80 ± 0.82) days] (P lt; 0.05), and in group C than in group B (P lt; 0.05). At 7 days after transplantation, the blood leukocyte number was (6.32 ± 0.45) × 109/L in group A, (7.45 ± 0.52) × 109/L in group B, and (6.35 ± 0.39)× 109/ L in group C, and it was significantly more in group B than in groups A and C (P lt; 0.05). The macrophage activation rate of the abdominal fluid was 6.87% ± 2.40% in group A, 7.84% ± 0.44% in group B, and 15.98% ± 2.87% in group C; group C was significantly higher than groups A and B (P lt; 0.01). ELISA results showed that there was no significant difference in the concentrations of IL-4 among 3 groups (P gt; 0.05). Compared with group B, the concentrations of IL-17 and IFN-γ were significantly reduced in group C (P lt; 0.05), while the concentration of IFN-γ was significantly increased in group B when compared with group A (P lt; 0.05). RT-PCR results showed that there were significant differences in the expressions of IL-4, IL-17, and IFN-γ mRNA between groups B, C and group A (P lt; 0.05); the expressions of IL-4 and IFN-γ mRNA were significantly lower in group C than in group B (P lt; 0.05). Conclusion Human PMSCs transplantation can suppress the acute immunological rejection in allogeneic skin transplantation. The possible mechanism may be partially related to the inhibitory effect on the secretion of IL-17 and IFN-γ.