ObjectiveTo evaluate the application of portosystemic shunt after subcutaneous transposition of the spleen (STS) to orthotopic liver transplantation (OLT) in the rat. MethodsOne hundred and eighty Wistar rats were randomly divided into the group of orthotopic liver transplantation after portosystemic shunts by subcutaneous transposition of the spleen (STS+OLT group) and the group of orthotopic liver transplantation (OLT group). The two groups were further divided into A, B, C subgroups in the light of duration of anhepatic phase (phases were respectively less than 25 min, around 35 min and 45 min).There were fifteen rats in each subgroup. At the described intervals, blood samples were collected from the peripheral and portal vein for testing ALT, pH and endotoxin levels. The survival rates were also observed. Results The ALT value of all animals basically returned to normal levels on the 7th postoperative day in the STS+OLT group and the OLT A subgroup, but in OLT B subgroup, ALT was still remarkably elevated on the 7th postoperative day (P＜0.01), and returned to normal levels on the 30th postoperative day. The pH values and endotoxin levels from the portal vein of all animals in STS+OLT groups and OLT A subgroup had no significant difference (Pgt;0.05) at the beginning, the end of the anhepatic phase and at the time of reperfusion for 30 min. But in the OLT B and C groups, the pH values and endotoxin levels were significantly higher at the end of anhepatic phase and reperfusion for 30 min than those in the beginning of anhepatic phase (P＜0.01). The survival rates at postoperative different time points in both B and C subgroup of the OLT group were significantly lower than those in STS+OLT group animals (P＜0.05).Conclusion The portosystemic shunt by subcutaneous transposition of the spleen can notably improve both the success rate of the OLT procedure and the postoperative survival rate in the rat.
Objective To analysis the common reasons for failure in orthotopic liver transplantation during preliminary experiment and propose the preventive. Methods One hundred and twenty cases in preliminary experiment using modified Kamada “two-cuff” method of orthotopic liver transplantation were retrospectively analyzed. Results The causes of failure included： lengthening of anhepatic phase (66 cases)， failed anastomosis of suprahepatic inferior vena cava (61 cases)， failed anastomosis of infrahepatic inferior vena cava (17 cases), failed anastomosis of portal vein (12 cases), unsatisfied anesthesia (8 cases). Succeed in 21 cases (17.50%, 21/120). Conclusion Improve the microsurgical operation techniques, particularly the anastomosis of suprahepatic inferior vena cava, can increase the success rate in orthotopic liver transplantation.
To explore the role of cell apoptosis in denervated skeletal muscle atrophy in rats and the effect of losartan on it. Methods Forty-two Sprague Dawley rats were randomly divided into 3 groups: group I (n =14, normal control group), group II (n =14, denervated group) and group III (n =14, losartan group). The rats were not treated in group I, and were made denervated gastrocnemius models in groups II and III. In group III, the rats were treated with losartan 10 mg /kg• d by gavage and with normal sal ine in groups I and II. After 4 weeks, gastrocnemius mass to body mass ratio (GAS/BM) served as the degree of muscle atrophy. Apoptotic cells in gastrocnemius were stained in situ by using TUNEL. Gastrocnemius Bcl-2 and Bax protein were quantified by immunohistochemistry and Western blot. Bax /Bcl-2 served as the degree of apoptosis. Results The ratio of apoptosis was higher in group II than that in group I (11.32% ± 4.51% vs 0.56% ± 0.21%, P lt; 0.05). The ratio of apoptosis was lower in group III than that in group II (7.21% ± 2.05% vs 11.32% ± 4.51%, P lt; 0.05). The atrophy of skeletal muscle（GAS/BM） in group II was more serious than that in group I (11.68 ± 1.98 vs 12.86 ±0.74, P lt; 0.05), there was no significant difference between group III and group II (12.11 ± 0.65 vs 11.68 ± 1.98, P gt; 0.05). The expression of Bcl-2 in group II (18.3% ± 4.9%) was significantly lower than that in group I (27.5% ± 2.8%) and group III (25.5% ± 3.5%); there was no significant difference between group III and group I (P gt; 0.05). The expression of Bax in group II (24.1% ± 3.1%) was significantly higher than that in group I (22.1% ± 3.6%) and group III (21.7% ± 2.3%); there was no significant difference between group III and group I (P gt; 0.05). Western blot results showed that: the expressions of Bcl-2 were 122.5 ± 14.6 in group II, 135.3 ± 6.2 in group I and 139.2 ± 16.2 in group III; showing significant diffeerences between group II and group I, between group III and group II (P lt;0.05). The expressions of Bax were 107.1 ± 15.8 in group II, 89.3 ± 8.4 in group I, and 94.2 ± 9.5 in group III; showing significant diffeerences between group II and group I, between group III and group II (P lt; 0.05). There was no significant difference in the expression of Bcl-2 and Bax between group Ⅲ and group I (P gt; 0.05). Conclusion Cell apoptosis plays an important role in denervated skeletal muscle atrophy in rats and may be one of the factors causing skeletal muscle atrophy. Losarton can decrease skeletal muscle cell apoptosis through regulating the ratio of Bax / Bcl-2.
【摘要】 目的 观察在不同剂量乙醇作用下大鼠下丘脑和脊髓神经细胞P物质的表达情况和扫描电子显微镜（SEM）下神经细胞的形态学变化，探讨乙醇作用下大鼠行为学改变的相关机制。 方法 通过福尔马林实验观察大鼠在不同剂量乙醇及时间作用下行为学的改变；采用免疫组织化学技术检测不同剂量乙醇作用下大鼠脊髓和下丘脑神经细胞中P物质的表达，通过扫描电子显微镜观察神经细胞的形态学变化。 结果 乙醇灌胃后0～2 h大鼠舔足次数有不同程度的变化，组间比较差异有统计学意义（Plt;0.05），灌胃2 h大鼠下丘脑和脊髓P物质表达程度与乙醇剂量有相关关系，扫描电子显微镜下各组大鼠的神经细胞形态学变化显著。 结论 急性乙醇中毒可引起大鼠对疼痛反应的变化，其程度与乙醇剂量和作用时间有关，大鼠下丘脑和脊髓神经细胞中P物质的表达强度与乙醇剂量和作用时间有关。【Abstract】 Objective To observe the expression of substance P(SP)in the hypothalamus and spinal cord nerve cells of rats with different concentrations of ethanol, and to observe the morphological changes of nerve cells by scanning electron microscopic(SEM) for elucidating the mechanism of ethological changes effected with ethanol. Methods Ethological changes were detected through the formalin test; SP expressions in the hypothalamus and the spinal cord were evaluated with immunohistochemistry technology, and the morphological changes of nerve cells were observed by SEM. Results The frequency of licking foot changed when the rats were gavaged with different concentrations of ethanol among zero to two hours, the difference between two groups was statistical signifcant (Plt;0.05). The expression level of SP and the morphological changes of nerve cells in hypothalamus and spinal cord had relationship with the ethanol concentration. Conclusions Acute alcoholism could cause pain dysfunction in rats. The frequency of licking foot of rats is correlated to the role of the time closely. The expression intensity of SP in the hypothalamus and the spinal cord nerve cells are correlated to the concentration of ethanol closely.
【Abstract】Objective To investigate the effect of donor blood transfusion on inducing pancreatic allograft tolerance in outbred rat model. Methods Wistar male rats were used as blood and pancreas donor, and diabetic recipients. One ml of donor blood injected into abdomen of diabetic recipients on the day of transplantation and azathioprine given 2 days pretransplant and continued for three days. Results Pancreas allograft survival was significantly prolonged (28 to 112 days, media survival time 64.2 days). One ml of donor blood alone injected into the abdomen and azathioprine given alone 2 days pretransplant did not improve allograft survival (media survival time 9.8 vs 10.2 days). Conclusion Donor blood injected on the day of transplantation and a 3 days course of azathioprine started 2 days pretransplant have b synergism in inducing long term graft survival in this rat model.
Objective To study the effect of recombinant lentiviral vector mediated human hepatocyte growth factor (hHGF) gene-modified bone marrow mesenchymal stem cells (BMSCs) on the immunological rejection after allograft l iver transplantation in rats, and to reveal the mechanism of immune tolerance. Methods Eight male Sprague Dawley (SD)rats of clean grade (aged 3 to 4 weeks, weighing 75-85 g) were selected for the isolation and culture of BMSCs; 64 adult male SD rats of clean grade (weighing 200-250 g) were used as donors; and 64 adult male Wistar rats of clean grade (weighing 230-280 g) were used as receptors. After establ ishing a stable model of rat allogeneic l iver transplantation, 1 mL sal ine, 2 ×106/mL of BMSCs 1 mL, 2 × 106/mL of BMSCs/green fluorescent protein 1 mL, and 2 × 106/mL of BMSCs/hHGF 1 mL were injected via the portal vein in groups A, B, C, and D respectively. Then the survival time of the rats was observed. The hepatic function was determined and the histological observation of the l iver was performed. The hHGF mRNA expression was detected by RT-PCR, the level of cytokine including hHGF, interleukin 2 (IL-2), IL-4, IL-10, and interferon γ (IFN-γ) by ELISA assay, the level of apoptosis by TUNEL method, and the expression level of prol iferating cell nuclear antigen (PCNA) by immunohistochemical method. Results The survival time of group D was significantly higher than that of groups A, B, and C (P lt; 0.01); the survival time of groups B and C was significantly higher than that of group A (P lt; 0.01), but there was no significant difference between group B and group C (P gt; 0.05). RT-PCR demonstrated the transcription of hHGF mRNA in the grafts of group D; the serum cytokine hHGF reached to (6.2 ± 1.0) ng/mL. Compared with groups B and C, group D exhibited significant inhibitory effect, significantly improved l iver function, and showed mild acute rejection. In addition, the levels of cytokine IL-2 and IFN-γ decreased; the levels of cytokine IL-4 and IL-10 increased; the level of apoptosis reduced; and the expression level of PCNA increased. Except for the expression of IL-4 (P gt; 0.05), there were significant differences in the other indexes between group D and groups B, C (P lt; 0.05). Conclusion BMSCs/hHGF implanting to rat l iver allograft via portal vein can induce immune tolerance. Compared with injection of BMSCs alone, BMSCs/hHGF treatment can alleviate acute rejection and prolong the survival time significantly. The immunosuppressive effect of BMSCs/hHGF is correlated with Th2 shifts up of Th1/Th2 shift, reduced apoptosis, promoted l iver regeneration.
ObjectiveTo provide the reliable model in the rat for the study of liver transplantation.MethodsA surgical experience with one hundred and fifty orthotopic liver transplants in rats was reviewed,meanwhile a simple method with biliary extradrainage as well as bile collection was introduced.Results The operative successful rate was 93.8%（91/97）,the survival rate after one week was 85.9%(55/64),the recipient underwent an anhepatic period of 19 min,the operative time of donor and recipient were 37 min and 56 min,respectively.ConclusionThe result indicates that the model is reliable in the study of liver transplantation.The hard work and meticulous surgical performance are the key to successful operation.
Objective To investigate the effect of stretch on long non-coding RNA taurine upregulated gene 1 (TUG1)-mediated miR-545-3p/cannbinoida receptor 2 (CNR2) pathway regulating bone regeneration in the distraction area of rats during distraction osteogenesis. MethodsThirty-six 10-week-old male Sprague Dawley rats were randomly divided into 3 groups (n=12 in each group): group A (femoral fracture+injection of interfering RNA), group B (distraction osteogenesis+injection of interfering RNA), and group C (distraction osteogenesis+injection of TUG1). Groups A and B were injected with 60 μg of interfering RNA at the beginning of incubation period (immediate after operation), the beginning of distraction phase (7 days after operation), and the end of distraction phase (21 days after operation), and group C was injected with 60 μg of synthetic TUG1 in vivo interfering sequence at the same time. The general situation of rats in each group was observed during the experiment. The mineralization of fracture space or distraction area was observed by X-ray films at 21, 35, and 49 days after operation. At 49 days after operation, the samples of the distraction area were taken for HE staining to observe the mineralization, and real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the expressions of osteoblast-related genes such as TUG1, miR-545-3p, CNR2, alkaline phosphatase (ALP), osteocalcin (OCN), and osteopontin (OPN). Blood samples were collected from the abdominal aorta of the rats, and the expressions of ALP and C terminal telopeptide of type Ⅰ (CTX-Ⅰ) protein were detected by ELISA assay.Results The results of X-ray film and HE staining observations showed that osteogenesis in group C was superior to groups A and B at the same time point. The results of qRT-PCR showed that the relative mRNA expressions of TUG1, CNR2, ALP, OCN, and OPN in group C were significantly higher than those in group A and group B, and the relative mRNA expression of miR-545-3p in group C was significantly lower than that in group A and group B (P<0.05). The relative mRNA expressions of TUG1 and ALP in group B were significantly higher than those in group A, and the relative mRNA expression of miR-545-3p in group B was significantly lower than that in group A (P<0.05). There was no significant difference in the relative mRNA expressions of CNR2, OCN, and OPN between group A and group B (P>0.05). The results of ELISA showed that the expressions of ALP and CTX-Ⅰ protein were significantly higher in group C than in group A and group B, and in group B than in group A (P<0.05). ConclusionUnder the action of stretch, the expression of TUG1 in the femoral distraction area of rats increases, which promotes the expression of CNR2 by inhibiting the expression of miR-545-3P, which is helpful to the mineralization of the extension area and osteogenesis.
ObjectiveTo study the feasibility of the establishment of transplantated hepatic cancer models in SD rats. MethodsThe male weaning SD rats were inoculated and transgenerated from the celiar transplantation tumor rats (W2562k) by intraperitoneal injection of abdominal fluid of tumor cells, another weaningSD rats were prepared for the models of solid tumor by subcutaneous inoculation of the abdominal fluid of tumor cells. The solid tumors were cut to pieces of 0.2 cm×0.2 cm×0.3 cm,which were implanted into the liver of the adult male SD rats, and the transplantation hepative tumor models were established. After 7 days, the volume and weight of tumor mass was measured. ResultsThe successful rate of model was 100% (82/82), the natural extinctive rate was 0 (0/82), the successful rate of inoculation was 100% (15/15). ConclusionSD rats are economical, the successful rate of model was high, the natural extinctive low, so the SD rats are ideal selection in establishing transplantation liver cancer model.