An improved animal model of orthotopic liver transplantation in swine

Objective: To establish a swine model of orthotopic liver transplantation (OLT) which has high standardization, superior reproducibility and stability. Methods: The rate of success, reproducibility and stability were investigated on the modification of OLTs in closed miniature swine with series of improvements. Results: 20 OLTs were performed on the basis of improvements in experimental animals, surgical procedures and operative monitorings. The mean operation time and anhepatic phase was (181±25.8) and (28.4±3.2) min respectively, which were significantly shorter than those of the previous reports. Liver function of the animals recovered shortly after operation. One-week survival rate was 90%, and 15 animals survived more than 1 month. The incidence of vascular and biliary complications was lower in animals with long-term survival. Conclusion: The improved animal model of OLTs in swine is easy to operate with high standardization and rate of success, superior reproducibility and stability. It is an ideal model for series studies related to liver transplantation in big animals.

Improved two-cuff technique for orthotopic liver transplantation in rat

BACKGROUND: The first orthotopic liver transplantatio in rat (ROLT) was reported by Lee in 1973. Kamada inno vatively applied cuff technique to ROLT in 1979. However the operative procedures were highly demanding and th operative mortality was relatively high. The purpose of thi study was to improve the model of ROLT, simplify opera tive procedures, and enhance the successful rate of opera tion. METHODS: Orthotopic liver transplantation was per formed in 160 Wistar rats by improved two-cuff technique The portal vein between donor and recipient was anasto mosed with the cuff technique. The same method was use to anastomose the infrahepatic vena cava. The suprahepati vena cava and the hepatic artery were anastomosed by m crovascular suturing and the bile duct was anastomosed en to end by a Teflon catheter. RESULTS: The average time for donor operation, graf preparation and anhepatic phase was 31 minutes, 14 mi nutes and 13 minutes, respectively. The anastomosis tim for the suprahepatic vena cava, portal vein, infrahepatic ve na cava, hepatic artery and bile duct was 7 minutes, 2 mi nutes, 2 minutes, 8 minutes and 1 minute, respectively The main causes for operative mortality were pneumotho rax, anesthesia, air embolism and massive bleeding, an the successful rate of operation was 92.5%. The causes fo death after operation were stoma bleeding, infection, b liary obstruction and graft failure. CONCLUSION: The improved two-cuff technique can re duce operative mortality, enhance survival rate, and serv as an ideal method for the establishment of animal model o ROLT.

Using Tree Shrews (Tupaia belangeri)as a Novel Animal Model of Liver Transplantation

Liver transplantation (LT)is most effective and promising approach for end-stage liver disease.However,there remains room for further improvement and innovation,for example, to reduce ischemic reperfusion injury,transplant rejection and immune tolerance.A good animal model of LT is essential for such innovation in transplant research.Although rat LT model has been used since the last century,it has never been an ideal model because the results observed in rat may not be applied to human because these two species are genetically distinct from each other.In this study,we for the first time performed LT using the tree shrew (Tupaia belangeri),a species in the Order Scandentia which is closely related with primates,and evaluated the possibility to adopt this species as a new model of LT.We performed LT on 30 animals using the two-cuff technique, examining the success rate,the survival rate and the immunological reaction.The recipient operation time was 60 min averagely,and we limited the time of the anhepatic phase within 20 min.Twenty-seven (90%)of the animals survived for at least 3 days after the transplantation. Thirteen animals that did not receive any immunosuppressive drug died in 8 days mostly because of acute rejection effect (n=9),similar to the reaction in human but not in experimental rat.The rest 14 animals that were given rapamycin survived significantly longer (38 days)and half of them survived for 60 days until the end of the study.Our results suggest that performing LT in tree shrews can yield high success rate and high survival rate.More importantly,the tree shrews share similar immunological reaction with human.In addition,previous genomics study found that the tree shrews share more proteins with human.In sum,the tree shrews may outperform the experimental rats and could be used as a better and cost-effective animal model for LT.

Modified arterialization of orthotopic liver transplantation in a mouse model

BACKGROUND: With the establishment of genetically modified and gene knock-out models, the mouse has become an important animal model for liver transplantation. We examined hepatic rearterialization after liver transplantation in a mouse model. METHODS: Orthotopic liver transplantation was performed in 70 mice and sham-operation was performed in a control group of 40 mice. Based on the 'two-cuff' method, a continuous suture approach was applied to the suprahepatic inferior vena cava and a cuff approach to the portal vein and the infrahepatic inferior vena cava. A biliary stent was inserted into the bile duct. The hepatic artery was reconstructed with end-to-side anastomosis. The survival rate of recipients was monitored at 24 hours, one week, and one month after the operation. Liver function and morphology were evaluated one month postoperatively. RESULTS: Postoperative survival rates were 94.3% at 24 hours, 91.4% at one week, and 85.7% at one month. No significant difference was seen between the experimental and control groups in liver function. The hepatic tissue preserved normal structure. CONCLUSION: Owing to its high survival rate and stability, this surgical approach is ideal for establishing an orthotopic liver transplantation mouse model with hepatic artery reconstruction. (Hepatobiliary Pancreat Dis Jut 2010; 9: 264-268)

Establishment of a new pig model for auxiliary partial orthotopic liver transplantation

AIM: To establish a new pig model for auxiliary partial orthotopic liver transplantation (APOLT).METHODS: The liver of the donor was removed from its body. The left lobe of the liver was resected in vivo and the right lobe was used as a graft. After the left lateral lobe of the recipient was resected, end-to-side anastomoses of suprahepatic inferior vena cava and portal vein were performed between the donor and recipient livers,respectively. End-to-end anastomoses were made between hepatic artery of graft and splenic artery of the host.Outside drainage was placed in donor common bile duct.RESULTS: Models of APOLT were established in 5 pigs with a success rate of 80%. Color ultrasound examination showed an increase of blood flow of graft on 5th d compared to the first day after operation. When animals were killed on the 5th d after operation, thrombosis of hepatic vein (HV) and portal vein (PV) were not found. Histopathological examination of liver samples revealed evidence of damage with mild steatosis and sporadic necrotic hepatocytes and focal hepatic lobules structure disorganized in graft. Infiltration of inflammatory cells was mild in portal or central vein area. Hematologic laboratory values and blood chemical findings revealed that compared with group A (before transplantation), mean arterial pressure (MAP), central venous pressure (CVP), buffer base (BB), standard bicarbonate (SB) and K+ in group B (after portal vein was clamped) decreased (P＜0.01). After reperfusion of the graft, MAP, CVP and K+ restored gradually.CONCLUSION: Significant decrease of congestion in portal vein and shortened blocking time were obtained because of the application of in vitro veno-venous bypass during complete vascular clamping. This new procedure,with such advantages as simple vessel processing, quality anastomosis, less postoperative hemorrhage and higher success rate, effectively prevents ischemia reperfusion injury of the host liver and deserves to be spread.

Details determining the success in establishing a mouse orthotopic liver transplantation model

Liver transplantation(LT)is currently the only effective treatment option for endstage liver disease.The importance of animal models in transplantation is widely recognized among researchers.Because of the well-characterized mouse genome and the greater diversity and availability of both genetically modified animals and research reagents,mouse orthotopic LT(MOLT)has become an ideal model for the investigation of liver biology,tissue injury,regulation of alloimmunity and tolerance induction,and the pathogenesis of specific liver diseases.However,due to its complicated and technically demanding procedure,the model has merely been used by only a few research groups in the world for years.For a new learner,training lasting at least a couple of months or even years is required.Most of the investigators have emphasized the importance of elaborate techniques and dedicated instruments in establishing a MOLT model,but some details are often neglected.The nontechnical details are also significant,especially for researchers who have little experience in mouse microsurgery.Here,we review and summarize the crucial technical and nontechnical details in establishing the model of MOLT based on scientific articles and our experience in six aspects:animal selection,anesthesia,perioperative management,organ procurement,back-table preparation,and implantation surgery.We aim to enable research groups to shorten the learning curve and implement the mouse LT procedure with high technical success.

Study of heteroserum-induced rat liver fibrosis model and its mechanism

AIM To investigate the morphological changes in the process of heteroserum induced rat liver fibrosis and the mechanism of fibrogenesis of this model. METHODS A model of heteroserum induced rat liver fibrosis was established by intraperitoneal injection of porcine serum. In addition to the observation of the morphological changes of this model, the infiltration of eosinophils and mast cells were measured quantitatively and the deposition of IgG and complement C 3 was detected by immunofluorescence. RESULTS The rat liver fibrosis was induced successfully at the end of the 8th week after the injection of heteroserum. Besides the increase of hepatic stellate cells (HSC) during the process of liver fibrosis, proliferation and activation of primary mesenchyma cells (PMCs) were also found. In the early stage, the infiltration of eosinophils and mast cells was significantly increased and the deposition of IgG and complement C 3 was positive in the portal tracts and septa, while gradually reduced after the injection was stopped. CONCLUSIONS This model is suitable for the research on liver fibrogenesis; the pathogenesis of this model may be related with the allergen induced late phase reaction (LPR) caused by the injection of heteroserum, and the HSCs and the PMCs are important sources of ECM producing cells.

Biliary tract injury caused by different relative warm ischemia time in liver transplantation in rats

BACKGROUND: There is a controversy over the degree of liver and biliary injury caused by the period of secondary warm ischemia. A liver autotransplantation model was adopted because it excludes the effects of infection and immunological rejection on bile duct injury. This study was undertaken to assess biliary tract injury caused by relative warm ischemia (secondary warm ischemia time in the biliary tract) and reperfusion. METHODS: One hundred and two rats were randomly divided into 5 groups: group I (control); groups 11 to V, relative warm ischemia times of 0 minute, 30 minutes, I hour and 2 hours. In addition to the levels of serum alkaline phosphatase, and total bilirubin, pathomorphology assessment and TUNEL assay were performed to evaluate biliary tract damage. RESULTS: Under the conditions that there were no significant differences in warm ischemia time, cold perfusion time and anhepatic phase, group comparisons showed statistically significant differences. The least injury occurred in group H (portal vein and hepatic artery reperfused simultaneously) but the most severe injury occurred in group V (biliary tract relative warm ischemia time 2 hours). CONCLUSIONS: Relative warm ischemia is one of the factors that result in bile duct injury, and the relationship between relative warm ischemia time the bile injury degree is time-dependent. Simultaneous arterial and portal reperfusion is the best choice to avoid the bile duct injury caused by relative warm ischemia. (Hepatobiliary Pancreat Dis Int 2009; 8: 247-254)

Comprehensive and innovative techniques for livertransplantation in rats: A surgical guide

AIM: To investigate our learning curves of orthotopic liver transplantation (OLT) in rats and the most important factor for successful surgery. METHODS: We describe the surgical procedures for our rat OLT model, and determined the operator learning curves. The various factors that contributed to successful surgery were determined. The most important surgical factors were evaluated between successful and unsuccessful surgeries.RESULTS: Learning curve data indicated that 50 cases were required for operator training to start a study. Operative time, blood loss, warm ischemic time, anhepatic phase, unstable systemic hemodynamic state, and body temperature after surgery significantly affected surgery success by univariate analysis, while the anhepatic phase was the most critical factor for success by multivariate analysis. CONCLUSION: OLT in rats is the only liver transplantation model that provides clinically relevant and reliable results. Shortened anhepatic phase is key to success in this model.

Enteral feeding of glycyl-glutamine dipeptide improves the structure and absorptive function of the small intestine after allogenetic liver transplantation in rats

BACKGROUND: Recipients of liver transplantation could have postoperative structural injury and declined absorptive function in the gastrointestinal tract. Glutamine (Gln) is a special nutrient of small intestinal mucosa and of various kinds of cells proliferating rapidly. But Gln could form a kind of poisonous pyroglutamic acid in water solution, which is the limitation of Gln in clinical practice. Glycyl-glutamine (Gly-Gln) is highly soluble and can be hydro-lyzed to release glutamine. This study was undertaken to observe the effect of Gly-Gln dipeptide by enteral feeding on the intestinal structure and absorptive function after allogenetic liver transplantation in rats. METHODS: Twelve male inbred Lewis rats were selected randomly as donors, and 24 male inbred BN rats as recipients of allogenetic liver transplantation. The recipients were also randomly divided into two groups; control group (ALA group, n = 12 ) and experimental group ( GLN group , n =12). In each group, 6 normal BN rats were sampled as the normal parameter on the 3rd preoperative day. The 6 recipients in the control group received alanine 0. 6 g/kg daily for 3 days before operation and 7 days after operation by gastric perfusion, and the 6 recipients in the experimental group were given Gly-Gln 0.6 g/kg daily the same way. The 12 BN recipients underwent 3-day fasting (free access to water with 0. 23% sodium chloride) and ortho-topic liver transplantation in aseptic conditions and were given subcutaneous injection of CsA 2 mg/kg daily after the operation. The 12 BN recipients were sampled on the 8th postoperative day. All of the 24 BN rats were subjected to examination of mucosal structure, activities of Na + -K + - ATP and disaccharidase, and D-xylose absorption test. RESULTS: The 12 BN recipients were alive after liver transplantation. On the 3rd preoperative day, mucosal structure , activities of Na + -K -ATP and disaccharidase and D-xylose absorption in the two groups were not significantly different. On the 8th postoperative day, the parameters of the two groups were markedly changed compared with those on the 3rd preoperative day. However, the parameters of GLN group were remarkably higher than those of ALA group. CONCLUSION: Enteral feeding of Gly-Gln could improve the structure and absorptive function of the small intestine after liver transplantation in rats.

Intravenous administration of glutathione protects parenchymal and non-parenchymal liver cells against reperfusion injury following rat liver transplantation

AIM:To investigate the effects of intravenous administration of the antioxidant glutathione (GSH) on reperfusion injury following liver transplantation. METHODS:Livers of male Lewis rats were transplanted after 24 h of hypothermic preservation in University of Wisconsin solution in a syngeneic setting.During a 2-h reperfusion period either saline (controls,n=8) or GSH (50 or 100 μmol/(h·kg),n=5 each) was continuously administered via the jugular vein. RESULTS:Two hours after starting reperfusion plasma ALT increased to 1 457±281 U/L (mean±SE) in controls but to only 908±187 U/L (P<0.05) in animals treated with 100 μmol GSH/(h·kg).No protection was conveyed by 50μmol GSH/(h·kg).Cytoprotection was confirmed by morphological findings on electron microscopy:GSH treatment prevented detachment of sinusoidal endothelial cells (SECs) as well as loss of microvilli and mitochondrial swelling of hepatocytes.Accordingly,postischemic bile flow increased 2-fold.Intravital fluorescence microscopy revealed a nearly complete restoration of sinusoidal blood flow and a significant reduction of leukocyte adherence to sinusoids and postsinusoidal venules.Following infusion of 50μmol and 100 μmol GSH/(h·kg),plasma GSH increased to 65±7 mol/L and 97±18 mol/L,but to only 20±3 mol/L in untreated recipients. Furthermore,plasma glutathione disulfide (GSSG) increased to 7.5±1.0 mol/L in animals treated with 100μmol/(h·kg) GSH but infusion of 50μmol GSH/(h·kg) did not raise levels of untreated controls (1.8±0.5 mol/L vs 2.2±0.2 mol/L). CONCLUSION:Plasma GSH levels above a critical level may act as a “sink” for ROS produced in the hepatic vasculature during reperfusion of liver grafts.Therefore,GSH can be considered a candidate antioxidant for the Drevention of reperfusion injury after liver transplantation,in particular since it has a low toxicity in humans.

Experimental models of non-alcoholic fatty liver disease in rats

Non-alcoholic fatty liver disease(NAFLD)is the most common chronic liver disease in the Western world,and it persists at a high prevalence.NAFLD is characterised by the accumulation of triglycerides in the liver and includes a spectrum of histopathological findings,ranging from simple fatty liver through non-alcoholic steatohepatitis(NASH)to fibrosis and ultimately cirrhosis,which may progress to hepatocellular carcinoma.The pathogenesis of NAFLD is closely related to the metabolic syndrome and insulin resistance.Understanding the pathophysiology and treatment of NAFLD in humans has currently been limited by the lack of satisfactory animal models.The ideal animal model for NAFLD should reflect all aspects of the intricate etiopathogenesis of human NAFLD and the typical histological findings of its different stages.Within the past several years,great emphasis has been placed on the development of an appropriate model for human NASH.This paper reviews the widely used experimental models of NAFLD in rats.We discuss nutritional,genetic and combined models of NAFLD and their pros and cons.The choice of a suitable animal model for this disease while respecting its limitations may help to improve the understanding of its complex pathogenesis and to discover appropriate therapeutic strategies.Considering the legislative,ethical,economical and health factors of NAFLD,animal models are essential tools for the research of this disease.

Orthotopic liver transplantation for giant liver haemangioma: A case report

In liver haemangiomas, the risk of complication rises with increasing size, and treatment can be obligatory. Here we present a case of a 46-year-old female who suffered from a giant haemangioma causing severe portal hypertension and vena cava compression, leading to therapy refractory ascites, hyponatremia and venostasis-associated thrombosis with pulmonary embolism. The patients did not experience tumour rupture or consumptive coagulopathy. Surgical resection was impossible because of steatosis of the non-affected liver. Orthotopic liver transplantation was identified as the only treatment option. The patient's renal function remained stable even though progressive morbidity and organ allocation were improbable according to the patient's lab model for end-stage liver disease(lab MELD) score. Therefore, non-standard exception status was approved by the European organ allocation network "Eurotransplant". The patient underwent successful orthotopic liver transplantation 16 mo after admission to our centre. Our case report indicates the underrepresentation of morbidity associated with refractory ascites in the lab MELD-based transplant allocation system, and it indicates the necessity of promptly applying for non-standard exception status to enable transplantation in patients with a severe clinical condition but low lab MELD score. Our case highlights the fact that liver transplantation should be considered early in patients with non-resectable, symptomatic benign liver tumours.

Animal models of ex vivo lung perfusion as a platform for transplantation research

Ex vivo lung perfusion(EVLP) is a powerful experimental model for isolated lung research. EVLP allows for the lungs to be manipulated and characterized in an external environment so that the effect of specific ventilation/perfusion variables can be studied independent of other confounding physiologic contributions. At the same time,EVLP allows for normal organ level function and real-time monitoring of pulmonary physiology and mechanics. As a result,this technique provides uniqueadvantages over in vivo and in vitro models. Small and large animal models of EVLP have been developed and each of these models has their strengths and weaknesses. In this manuscript,we provide insight into the relative strengths of each model and describe how the development of advanced EVLP protocols is leading to a novel experimental platform that can be used to answer critical questions in pulmonary physiology and transplant medicine.

Establishment and improvement of model of vascularized heart-thymus composite transplantation in rats

Objective To establish and improve the model of heart-thymus composite transplantation.Methods Vascularized both lobes of the thymus is transplanted heterotopically with the heart as a composite graft in rats.This technique was developed and assessed,and viability of the grafts was evaluated histologically.Results Donor operation costed 38.5±3.52 min,vascular anastomosis costed 25.0±3.28 min,operating successful rate was 90%,acute rejection was observed in SD-Wistar group,viable thymus with normal microarchitecture was maintained in Wistar-Wistar group.Conclusions The improved novel technique for combined heart-thymus transplantation is a valuable method for study of the role of thymus in transplantation immunity.

High incidence of biliary complications in rat liver transplantation:Can we avoid it?

MM： To investigate how to reduce the incidence of biliary complications in rat orthotopic liver transplantation. METHODS： A total of 165 male Wistar rats were ran- domly divided into three groups： Group A, orthotropic liver transplantation with modified ＂two-cuff＂ technique; Group B, bile duct was cut and reconstructed without transplantation; and Group C, only laparotomy was performed. Based on the approaches used for biliary reconstruction, Group A was divided into two sub-groups： A1 （n = 30）, duct-duct reconstruction, and A2 （n = 30）, duct-duodenum reconstruction. To study the influence of artery reconstruction on bile duct complication, Group B was divided into four sub-groups： B1 （n = 10）, duct-duct reconstruction with hepatic artery ligation, B2 （n = 10）, duct-duct reconstruction without hepatic artery ligation, B3 （n = 10）, duct-duodenum reconstruction with hepatic artery ligation, and B4 （n = 10）, duct-duodenum recon- struction without hepatic artery ligation. The samples were harvested 14 d after operation or at the time when significant biliary complication was found. RESULTS： In Group A, the anhepatic phase was 13.7 ＋ 1.06 min, and cold ischemia time was 50.5 ＋ 8.6 min. There was no significant difference between A1 and A2 in the operation duration. The time for biliary reconstruction was almost the same among all groups. The success rate for transplantation was 98.3% （59/60）. Significant differ- ences were found in the incidence of biliary complications in Groups A （41.7%）, B （27.5%） and C （0%）. A2 was more likely to have biliary complications than A1 （50% vs 33.3%）. B3 had the highest incidence of biliary complica- tions in Group B. CONCLUSION： Biliary complications are almost in- evitable using the classical ＂two cuff＂ techniques, and duct-duodenum reconstruction is not an ideal option in rat orthotopic liver transplantation.

A New Carotid Artery Transplantation Model of Rats

To establish a murine carotid artery transplantation model for the study of the chronic rejection, 80 rats were divided into two groups, an allotransplant （ACI-Lewis） group and an isotransplant （Lewis-Lewis） group （control group）. The donor carotid artery and the recipient carotid artery were anastomosed by using a polyethylene cuff （internal diameter： 0.7 mm, length： 3 mm）.The pathological changes of carotid artery transplant were observed 14, 28 and 56 days after the transplantation. The results showed that the model was successfully established in 95% of the animals. The chronic rejection-associated arteriosclerosis was induced 28 days after the transplantation. The new chronic rejection model of carotid artery by using cuff technique caused fewer traumas and was easy to make. The pathological changes of the transplant mimicked the chronic rejection-associated arteriosclerosis found in human transplant.

Rat Models of Fatty Liver due to Liver Depression and Spleen Deficiency

[Objectives] To establish animal models of fatty liver due to liver depression and spleen deficiency that were suitable for activity discovery of traditional Chinese medicine( TCM) compounds,efficacy evaluation,new drug research and development and correspondence between prescription and syndrome of TCM. [Methods] A syndrome score scale that was suitable for evaluating the rat models was established according to the evolvement rules of etiology and pathogenesis of TCM and the modern clinical pathological mechanism. At the same time,bifendate pills and Sanqi Zhigan pills were selected as drug counterevidence for the models. Rats in model groups were given different proportions of high-fat and low-protein fodder and different concentrations of alcohol every day,and intraperitoneally injected with porcine serum twice a week. Drug groups were given with bifendate pills and Sanqi Zhigan pills( 8. 1 mg/kg,and 2. 7 g/kg) respectively for 14 consecutive days.During the experiment,general status,weight,daily fodder intake and daily water intake of the animals were observed,and TCM syndromes were scored. After the experiment,the levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST),γ-glutamine transpeptidase( γ-GT),total cholesterol( TC),triglycerides( TG),low density lipoprotein-cholesterol( LDL-C),and high density lipoprotein-cholesterol( HDL-C) in serum were detected,and the pathological changes in liver tissue were observed. [Results] Compared with the control group,body weight,daily fodder and water intake of rats increased slowly in the model group,and the levels of ALT,AST,γ-GT,TC,TG and LDL-C increased significantly,while the level of HDL-C dropped. Pathological examination showed that steatosis and fat granule were observed in hepatocytes of rats in the model group. Behavioral observation found that main symptoms and minor symptoms of rats in the model group conformed to the syndrome manifestation of liver depression and spleen deficiency,which suggested that the three model groups were established successfully. Among them,three rats died in model group 1,and one rat died in model group 2. The manifestation of all the above lesions can be alleviated by drug counterevidence. [Conclusions]The animal model of fatty liver due to liver depression and spleen deficiency shows an obviously lower mortality and shorter duration and can be used for research of correlation between prescription and syndrome and its mechanism.

Therapeutic effect of Zijin capsule in liver fibrosis in rats

AIM To confirm the therapeutic effect of Zijin capsule on liver fibrosis in rat model. METHODS Model group: bovine serum albumin (BSA) Freund′s incomplete adjuvant 0 5ml was injected subdermally at d 1, d 15 , d 22 , d 29 and d 36 for primary sensitization. Seven days after the fifth injection, BSA antibody in the serum was detected by double agar diffusion method. Normal saline of 0 4ml was injected through cauda vein to BSA antibody positive rat twice a week for fifteen times. Traditional Chinese medicine (TCM) decoction group and Zijin capsule group: In the attack injection period, Chinese medicinal decoction or Zijin capsule was given ig, the others were the same as in the model group. NS was used in the control group. The collagen content of rat liver was determined by Bergman′s method and expressed as ±s . The liver pathological changes were divided into four grades and expressed as the avarage of the total rank sum. RESULTS The collagen content (mg/g) of the liver in the control group (7 2±1 9) was significantly lower than that in the other groups; it was higher in the model group (31 7±16 6) than that in the two therapeutic groups; and lower in Zijin capsule group (9 7±2 8) than that in the TCM decoction group (11 5±5 3). The pathological changes were more aggravated in the model group (37 4) than those in the two therapeutic groups; and more severe in the TCM decoction group (30.2) than in the Zijin capsule group (22.9) CONCLUSION The therapeutic effect of Zijin capsule on the model was confirmed.