Objective To explore the key technique of allogeneic whole pancreaticoduodenal transplantation (WPDT) in rats. MethodsWPDT model was established between Lewis rats as donors and Wistar rats (with type 1 diabetes mellitus) as recipients. End to side anastomosis was performed in abdominal aorta of donors and recipients. The portal vein of the graft was anastomosed with the recipients left renal vein by cuff technique. And side to side anastomosis was made between the graft duodenum and the host jejunum. ResultsForty-four of 50 rats were successfully performed WPDT. Amongthem, 8 rats died in postoperative 3 days, the survival time of residual 36 rats was 6-16 days, with an average of (10.45±3.30) days. The peak of death appeared on day 7-10 after operation. The typical acute rejection in pathological changes were observed on day 7. ConclusionSkilled microsurgical techniques and emphasis on details are important to establish WPDT model.
Objective To build animal models of keloid by method of tissue engineering and to discuss the feasibility of using it in clinical and lab researches. Methods Fibroblasts(FB) were isolated from keloids and cultured. The seventh and eighth generation of the cultured FBs were inoculated into the copolymers of polylactic acid and polyglycolic PLGA. After being cultured in rotatory cell culture system (RCCS)for 1 week,the FB was transplanted into athymic mice. The specimens were obtained 4 weeks and 8 weeks and examined histologically. Results All mice survived.The collagen patterns of all keloids were pressed in every specimen obtained 8 weeks. Fibrocytes andFB were observed in specimens by electronic microscope. There were abundent rough endoplasmic reticulum (RER) in FB, which indicated that FB’s capability of synthesizing and secreting collagen was preserved and the cellular characteristicwas remained. Conclusion There is a good affinity between PLGAand FB. The composition of PLGA and FB can form keloids in athymic mice,so that it deserves further researching and developing.
ObjectiveTo explore the feasibility of establishment of a artificial joint aseptic loosening mouse model by cobalt-chromium particles stimulation.MethodsTwenty-four 8-week-old male severe combined immunodeficient (SCID) mice were divided into experimental group (n=12) and control group (n=12). The titanium nail was inserted into the tibial medullary cavity of mouse in the two groups to simulate artificial joint prosthesis replacement. And the cobalt-chromium particles were injected into the tibial medullary cavity of mouse in experimental group. The survival of the mouse was observed after operation; the position of the titanium nail and the bone mineral density of proximal femur were observed by X-ray film, CT, and Micro-CT bone scanning; and the degree of dissolution of the bone tissue around the tibia was detected by biomechanical test and histological staining.ResultsTwo mice in experimental group died, and the rest of the mice survived until the experiment was completed. Postoperative imaging examination showed that there was no obvious displacement of titanium nails in control group, and there were new callus around the titanium nails. In experimental group, there was obvious osteolysis around the titanium nails. The bone mineral density of the proximal tibia was 91.25%±0.67%, and the maximum shear force at the tibial nail-bone interface was (5.93±0.85) N in experimental group, which were significantly lower than those in control group [102.07%±1.87% and (16.76±3.09) N] (t=5.462, P=0.041; t=3.760, P=0.046). Histological observation showed that a large number of inflammatory cells could be seen around the titanium nails in experimental group, while there was no inflammatory cells, and obvious bone tissue formation was observed in control group.ConclusionThe artificial joint aseptic loosening mouse model can be successfully established by cobalt-chromium particles stimulation.
Objective To provide a reliable experimental model for gastroesophageal reflux (GER) study. Methods Twenty Japan 5-month-old male rabbits wererandomly divided into two groups: group cardiomyotomy(n=10), group partial cardiomyectomy(n=10). The operations of cardiomyotomy and parital cardiomyectomy were performed in 2 groups respectively. All the animals underwent intraesophagealpH detection 1 week before operation and 4 weeks after operation. The mean changes of reflux ratios were compared between before operation and after operation.Results In gastroesophageal reflux ratio between before operation and after operation, there was no significant difference in group cardiomyotomy (1.98%±1.52% and 4.32%±2.39%, Pgt;0.05) and there was significant difference in group partialcardiomyectomy(1.56%±1.57% and 13.56%±3.27%, Plt;0.05). Conclusion The reliable experimental model of GER can be made with procedure of partial cardiomyectomy. It can be used in estimating the operative procedure of antireflux and is conducive to dynamic observation and study of esophagitis.
Objective To explore the feasibility of the Budd-Chiari syndrome model establishment in rat by using the inferior vena cava coarctation. Methods Fifty SD rats were randomly divided into experimental group and sham operation group, the laparotomy was performed after general anesthesia by intraperitoneal injection, and dissociated the inferior vena cava. In the experimental group, the vena cava was tightly ligated with silk thread according to partial portal vein coarctation, enclosing 23 G L-style blunt needle in the ligature to prevent complete obliteration. The diameter of the vena cava was set to about 80% of its normal size after removing the 23 G L-style blunt needle. The abdominal Doppler, liver function, blood routine examination, and liver biopsy were tested at different time (on week 1, 4, 8, and 12) after operation. Results The signs of inferior vena cava and primary hepatic venous obstruction, liver congestion and cirrhosis, ascites, hepatosplenomegaly, portal vein extension, and collateral patency occurred on week 4 in the experimental group. The levels of AST, ALT, AKP, TBIL, DBIL, and TBA in the experimental group were significantly higher than those in the sham operation group (P<0.05), and the WBC, PLT, RBC, HGB, and ALB in the experimental group was significantly lower than those in the sham operation group (P<0.05). Conclusion The inferior vena cava coarctation can be successfully used to establish a rat model of Budd-Chiari syndrome.
Objective To understand the advances in animal model and basic research of associating liver partition and portal vein ligation for staged hepatectomy (ALPPS), and to provide new ideas for basic research and clinical application of ALPPS. Methods The literatures on the basic research and animal models of ALPPS were analyzed and reviewed. Results By March 2018, there were 19 articles related to ALPPS animal models published, including 11 rat model articles, 4 mouse model articles, 2 pig model articles, 1 rabbit model article, and 1 sheep model article. These models of ALPPS were mainly simulated in normal liver background (16 articles), only 2 mouse model of colorectal liver metastasis and 1 rat model of ALPPS under the sclerotic liver background on Chinese article. In cases of rat’s models, portal blood flow deprivation of 20%–90% was finished by portal vein ligation, and the liver was localized and segmented according to the ischemic line and the ligaments of the liver, and the liver partition was mostly sutured and electrocoagulated to stop bleeding. In the above models, remnant liver hyperplasia was observed after surgery. The main causes of hyperplasia were serum cytokines-mediated [hepatocyte growth factor (HGF), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and so on] enhancement of proliferative gene, and secondly preservation of the portal vein lobes to increase blood volume and to accelerate liver proliferation. ConclusionsThe animal model is the main tool to study the safety of ALPPS and liver regeneration, but there are still few studies in the models with liver cirrhosis and liver tumors. The mechanism of liver regeneration after ALPPS is still unclear, and more basic experiments and clinical cases are needed for further study.
Objective To establish a reliable rats model of orthotopic liver transplantation with non-heart beating donors. Methods The model was established with modified double-cuff method. According to obtain pre-liver warm ischemia time experiencing non-heart-beat the rats were divided into 3 groups: 10 min (R10 group), 20 min (R20 group) and 30 min (R30 group), then one week survival after operation was compared in rats. Results The operative time of donor was 30 min approximately except warm ischemia time and the cold preservation time of donor liver was 1 h. The anastomotic time for suprahepatic vena cava was 12-22 min (mean 15 min). The anastomotic time for portal vein and infrahepatic vena cava was about 2 min and 1 min, respectively. The anhepatic phase sustained 14-24 min (mean 19 min). The operative time of receptor was 50-65 min (mean 60 min). Twelve rats died at 24 h after operation, which was considered as operative failure. The success rates of operation in R10 group, R20 group, and R30 group were 95% (19/20), 80% (16/20), and 65% (13/20), respectively. After one week the survival rate was 95% (18/19), 81% (13/16), and 54% (7/13), respectively. Conclusions Improved non-heart donor liver transplantation model of rat on the basis of Kamada’s “twocuff technique” acts as a good simulation in clinical non-heart-donor liver transplantation. This study showes that rat liver can tolerate warm ischemia time less than 30 min, the short-term survival after transplantation can reach satisfactory results. However, long-term survival requires further study.
Abstract: Objective To study the pathophysiological mechanism of the morphological change of immature pulmonary vessels in the piglet model of congenital heart defect with decreased pulmonary blood flow established with balloon atrial septostomy and pulmonary artery banding. Methods Twenty piglets at an age of one to two months were divided into three groups with random number table. For the control group (group C,n=6), small incisions were carried out on the right chest to produce a transient reduction in the pulmonary blood; for the lowmedium pulmonary artery stenosis group (group T1, n=7), the balloon dilator was delivered through the surface of the right atrium and septostomy and pulmonary artery banding were performed, and the systolic transpulmonary artery banding pressure (Trans-PABP) was controlled to be 20.30 mm Hg; For the severe pulmonary artery stenosis group (group T2, n=7), the same surgical procedures with group T1 were performed while TransPABP was controlled to be more [CM(159mm]than 3050 mm Hg.At 2 months after surgery respectively,a lung tissue of 1.0 cm×0.8 cm×0.8 cm from the lateral segment of the right middle lobe was taken out to be observed under optic microscope. The morphological change of the distal arterioles was detected. Furthermore, the content of vascular endothelial growth factor (VEGF) and matrix metalloproteinase2( MMP2) were also examined by the method of enzymelinked immunosorbent assay (ELISA). Results The model was successfully established in all the survival piglets of the group T1 and group T2. Two months after operation, the inner diameter of the pulmonary arterioles in group T1 was significantly higher than that in group C (82.89±10.72 μm vs.74.12±9.28 μm;t=-5.892, Plt;0.05), so as group T2 (85.47±5.25 μm vs.74.12±9.28 μm;t=-6.325, Plt;0.05); the number of arterioles per square centimeter (NAPSC) of group T1 was significantly lower than that of the group C (229.70±88.00 entries/cm 2 vs. 431.50±40.60 entries/cm2; t=39.526, Plt;0.05), so as group T2 (210.00±40.30 entries/cm2 vs. 431.50±40.60 entries/cm2; t=67.858, Plt;0.05). Two months after operation, the lung expression of MMP -2 and VEGF in group T1 was significantly lower than that in group C (58.30±19.60 ng/ml vs. 81.20±16.70 ng/ml, t=14.261, Plt;0.05; 17.80±3.00 pg/ml vs. 21.40±3.80 pg/ml, t=8.482, P<0.05), so does group T2 (42.10±15.20 ng/ml vs. 81.20±16.70 ng/ml, t=27.318, P<0.05; 12.30±3.20 pg/ml vs. 21.40±3.80 pg/ml, t=15.139, P<0.05). Conclusion Structural remodeling of pulmonary extracellular matrix is an important feature of the piglet model of congenital heart defect with decreased pulmonary blood flow. The arterioles show significant hypoplasia or degradation. Change in the structural proteins and cytokines during the reduction of blood in the lung is the key to structural remodeling.
A acute partial obstructive hepatocholangitis model by selective ligation and injection of E coli into left hepatic bile duct was successfully founded in rat. Using parameters including mortality, mitochondrial glutamic oxalacetic transaminase and ornithine carbamoytransferase activity, pathological observation and blood culture of bacteria, we evaluated the model. The authors emphasize that this models is superior to the wole-bile-duct-challenged cholangitis model, which is characterized by liver injury.
ObjectiveTo summarize the current researches and progress on experimental animal models of avascular necrosis of the femoral head. MethodsDomestic and international literature concerning experimental animal models of avascular necrosis of the femoral head was reviewed and analyzed. ResultsThe methods to prepare the experimental animal models of avascular necrosis of the femoral head can be mainly concluded as traumatic methods (including surgical, physical, and chemical insult), and non-traumatic methods (including steroid, lipopolysaccharide, steroid combined with lipopolysaccharide, steroid combined with horse serum, etc). Each method has both merits and demerits, yet no ideal methods have been developed. ConclusionThere are many methods to prepare the experimental animal models of avascular necrosis of the femoral head, but proper model should be selected based on the aim of research. The establishment of ideal experimental animal models needs further research in future.