ObjectiveTo summarize the application research progress of animal models of hepatocellular carcinoma at home and abroad in recent years. MethodThe relevant literature on animal models of hepatocellular carcinoma at home and abroad in recent years was reviewed. ResultsAt present, the common animal models of hepatocellular carcinoma mainly included spontaneous animal model, induced animal model, gene modified animal model, transplanted animal model, and humanized animal model. The etiology, pathogenesis, and pathophysiological changes of various animal models were different. The selection of animal models of hepatocellular carcinoma depended on the type and purpose of research. ConclusionsVarious animal models of hepatocellular carcinoma have their own advantages and disadvantages. Researchers should choose different animal models according to their own research purposes to achieve the expected experimental purposes, so that more valuable research results of animal models of hepatocellular carcinoma can be applied to clinical practice, and finally these research results can be truly applied to diagnosis and treatment of hepatocellular carcinoma.
ObjectiveTo observe the effect of using tungsten drills to prepare mouse knee osteochondral injury model by comparing with the needle modeling method, in order to provide an appropriate animal modeling method for osteochondral injury research.MethodsA total of 75 two-month-old male C57BL/6 mice were randomly divided into 3 groups (n=25). Mice in groups A and B were used to prepare the right knee osteochondral injury models by using needles and tungsten drills, respectively; group C was sham-operation group. The general condition of the mice was observed after operation. The samples were taken at 1 day and 1, 2, 4, and 8 weeks after modeling, and HE staining was performed. The depth, width, and cross-sectional area of the injury site at 1 day in groups A and B were measured, and the percentage of the injury depth to the thickness of the articular cartilage (depth/thickness) was calculated. Toluidine blue staining and immunohistochemical staining for collagen type Ⅱ were performed at 8 weeks, and the International Cartilage Research Society (ICRS) score was used to evaluate the osteochondral healing in groups A and B.ResultsAll mice survived to the completion of the experiment. HE staining showed that group C had normal cartilage morphology. At 1 day after modeling, the injury in group A only broke through the cartilage layer and reached the subchondral bone without entering the bone marrow cavity; the injury in group B reached the bone marrow cavity. The depth, width, cross-sectional area, and depth/thickness of the injury in group A were significantly lower than those in group B (P<0.05). At 1, 2, 4, and 8 weeks after modeling, there was no obvious tissue filling in the injured part of group A, and no toluidine blue staining and expression of collagen type Ⅱ were observed at 8 weeks; while the injured part of group B was gradually filled with tissue, the toluidine blue staining and the expression of collagen type Ⅱ were seen at 8 weeks. At 8 weeks, the ICRS score of group A was 8.2±1.3, which was lower than that of group B (13.6±0.9), showing significant difference (t=−7.637, P=0.000).ConclusionThe tungsten drills can break through the subchondral bone layer and enter the bone marrow cavity, and the injury can heal spontaneously. Compared with the needle modeling method, it is a better method for modeling knee osteochondral injury in mice.
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.
ObjectiveTo explore the method and feasibility of establishing patent ductus arteriosus (PDA) model in Bama miniature pig by using autologous jugular vein, and to provide a large animal model for the development of PDA occluder and the study of pulmonary hypertension associated with congenital heart disease. MethodsFive male Bama miniature pigs weighing about 45 kg were selected to gain the PDA model of the autogenous jugular vein, which was fixed by glutaraldehyde and anastomosed between the ascending aorta and the main pulmonary artery. The patency of PDA was confirmed by echocardiography and angiocardiography immediately and one week after the operation. Two animals were selected to undergo transcatheter closure of PDA via femoral vein 1 week after the operation, and the rest were euthanized to obtain PDA and lung tissue for pathological examination. ResultsThe PDA model was successfully established in all five animals with a success rate of 100.0%. Immediately and 1 week after the operation, echocardiography and angiography showed that PDA blood flow was unobstructed, and hematoxylin-eosin staining showed that PDA endothelialization was good. One week after the operation, two animals were successfully treated with transcatheter femoral vein occlusion. The pathological examination of lung tissue showed thickening of the intima and muscular layer of pulmonary arterioles, thickening of pulmonary interstitium and infiltration of neutrophils. ConclusionIt is safe and feasible to establish a large animal model of PDA by using autogenous jugular vein anastomosis between the ascending aorta and the main pulmonary artery. The model can be used for the development of PDA interventional occlusive devices and the pathophysiological study of congenital heart disease-related pulmonary hypertension.
ObjectiveTo explore the construction of heart preservation model of empty beating donor based on extracorporeal membrane oxygenation (ECMO). MethodsFrom January 2022 to August 2023, 20 Guangxi Bama miniature pigs weighing 25-30 kg were selected, half male and half female. Under general anesthesia and heparinization, a midline thoracotomy was performed. The pericardium was cut after freeing the anterior and posterior vena cavae, and a perfusion needle was inserted near the brachiocephalic artery in the ascending aorta, connected to a blood collection bag to collect 500-600 mL of blood. The anterior and posterior vena cavae were ligated, the aorta was blocked and perfused with HTK solution to stop the heart beating. The superior and inferior vena cavae were cut off, the right pulmonary vein was decompressed, the aorta and left and right pulmonary arteries and veins were cut off, and the whole heart was removed. An ECMO device was used to continuously perfuse a cardioprotective solution mainly composed of oxygenated warm blood, maintaining the isolated pig heart beating for 8 hours, monitoring (once/hour) ECMO perfusion parameters, blood gas indicators, perfusate electrolytes, detecting inflammatory factors, myocardial enzymes, myoglobin, and troponin levels. Myocardial tissue was taken for hematoxylin-eosin (HE) staining to observe myocardial cell damage and evaluate the quality of heart preservation. ResultsAmong the 20 isolated beating preservation pig hearts, 17 successfully resumed beating, 3 experienced ventricular fibrillation, resuscitated after intracardiac electrical defibrillation, and all 20 pig hearts successfully beat for 8 hours. There was no statistical difference in ECMO perfusion parameters, blood gas indicators, perfusate electrolytes, and inflammatory factors at each time point (P>0.05). There were statistical increases in myocardial enzymes, myoglobin, and troponin levels (P<0.05). HE staining results suggested that there was no severe myocardial damage. ConclusionECMO technology can be used for pig heart preservation with good results, and this study provides experimental evidence for improving heart preservation research in clinical heart transplantation.
Objective To observe the growth of orthotopic transplanted tumor in nude mice after stomatin-like protein 2 (SLP-2) expression decreased, and to further study the role of SLP-2 in the development and progression of esophageal squamous cell carcinoma. Methods Using RNA interference technique, esophageal squamous cell carcinoma cell lines with specific expression of SLP-2 and stable expression of luciferase were established. The healthy female nude mice with weight ranging from 19 to 22 g were randomly divided into 3 groups (n=12), 6 mice were used to establish subcutaneous xenografts, and the other 6 mice were used to establish the orthotopic transplanted tumor model (Group 1: cell infected with SLP-2-1 plasmid; group 2: cell infected with SLP-2-2 plasmid; group 3: cell infected with SHGFP plasmid). Index of the experiment end was weight loss and poor general situation in any mouse. Before the nude mice were sacrificed, the luciferase value of the tumor was detected by using in vivo imaging technique. After the nude mice were sacrificed, the primary tumor was removed for pathology examination. Results There was no significant difference in region of interest (ROI) value between the group 1 and group 2 (P=0.943). The ROI value for both groups 1 and 2 was significantly lower than that in the group 3 (P=0.002, P=0.000). The primary tumor infiltrated into the muscularis propria of esophageal was observed in all groups. Conclusion SLP-2 is involved in the development and progression of esophageal squamous cell carcinoma, and the decrease of SLP-2 expression can inhibit the growth of esophageal squamous cell carcinoma.
Objective To summarize the research progress of rodent models of secondary lymphedema (SL) and provide a reference for selecting appropriate animal models in SL research. Methods Recent literature on rodent SL models at home and abroad was comprehensively analyzed, summarizing model categories, development techniques, strengths, and weaknesses. Results Current research primarily utilizes rats and mice to establish SL models. The main model types include hind limb, forelimb, tail, and head/neck models. The hind limb model is the most frequently employed, typically requiring surgery combined with irradiation to induce stable chronic edema. Forelimb models primarily simulate upper limb lymphedema, but exhibit relatively rapid edema resolution. Tail models offer operational simplicity and are predominantly used for studying acute edema mechanisms and interventions; however, they demonstrate poor clinical relevance. Emerging head/neck models provide a valuable tool for investigating head and neck cancer-associated lymphedema. These models exhibit variations in lymphedema duration, degree of fibrosis, and edema incidences. Conclusion Existing models still fall short in faithfully replicating the chronicity, fibrosis, fat deposition, and complex microenvironment characteristic of human chronic lymphedema. Future research must integrate multidisciplinary approaches, optimize model construction strategies, and explore novel modeling approaches to more accurately mimic the human disease and advance SL prevention and treatment research.
This study aims to evaluate the ability of C-arm cone-beam CT to detect intracranial hematomas in canine models. Twenty one healthy canines were divided into seven groups and each group had three animals. Autologous blood and contrast agent (3 mL) were slowly injected into the left/right frontal lobes of each animal. Canines in the first group, the control group, were only injected with autologous blood without contrast agent. Each animal in all the 7 groups was scanned with C-arm cone-beam CT and multislice computed tomography (MSCT) after 5 minutes. The attenuation values and their standard deviations of the hematoma and uniformed brain tissues were measured to calculate the image noise, signal to noise ratio (SNR) and contrast to noise ratio (CNR). A scale with scores 1-3 was used to rate the quality of the reconstructed image of different hematoma as a subjective evaluation, and all the experimental data were processed with statistical treatment. The results revealed that when the density of hematoma was less than 65 HU, hematomata were not very clear on C-arm CT images, and when the density of hematoma was more than 65 HU, hematomata showed clearly on both C-arm CT and MSCT images and the scores of them were close. The coherence between the two physicians was very reliable. The same results were obtained with C-arm cone-beam CT and MSCT grades in measuring SD value, SNR, and CNR. The reasonable choice of density detection range of intracranial hematoma with C-arm cone-beam CT could be effectively applied to monitoring the intracranial hemorrhage during interventional diagnosis and treatment.
Objective To investigate the effect of chaiqin chengqi decoction (CQCQD) on serum lipid metabolism in experimental acute pancreatitis. Methods A total of 27 C57BL/6 mice were randomly divided into three groups (n=9 for each group). The mice in the acute pancreatitis model group (AP group) and the acute pancreatitis model + CQCQD treatment group (APQ group) received seven intraperitoneal injections of cerulein (50 µg/kg) at hourly intervals, while the mice in the control group (CON group) received phosphate-buffered saline injections at the same regimen of cerulein. Oral gavage of CQCQD (5.5 g/kg) or same volume of distilled water was commenced 1 h after the first cerulein injection for three times at intervals of 4 h for the APQ group and AP group, respectively. Animals were sacrificed 12 h after the first cerulein / phosphate-buffered saline injection for collecting serum and tissue samples. The levels of serum lipase and amylase, pancreatic histopathology assessment, as well as pancreatic myeloperoxidase activity, were used to assess the severity of acute pancreatitis and the efficacy of CQCQD. Additionally, serum lipid metabolites were analyzed in all groups. Results In comparison to the CON group, the mice in the AP group exhibited significant edema, inflammatory cell infiltration, necrosis of pancreatic tissues, as well as elevated levels of serum amylase, lipase, and pancreatic myeloperoxidase activity (P<0.05); in comparison to the AP group, inflammatory cell infiltration and necrosis of pancreatic tissue, as well as elevated level of serum amylase significantly reduced in the APQ group (P<0.05). A total of 319 lipid molecules were identified in serum, and 13 lipid metabolites were significantly increased in the AP group and successfully decreased in the APQ group, of which 9 were lyso-phosphatidylethanolamine (LPE) molecules involved in the glycerol phospholipid metabolic pathway. Further statistical analysis revealed that six of these LPE molecules could serve as potential biomarkers. Conclusions CQCQD ameliorated pancreatic injury and serum lipid metabolism disorder of acute pancreatitis model induced by cerulein and significantly improved the abnormal increase of serum LPE level. However, the role of LPE in acute pancreatitis and the underlying mechanisms of CQCQD on LPE metabolic pathways still need further study.
This study sought to investigate the in vivo antiviral effect of amantadine (AM) and biphenyl dimethyl dicarboxylate (DDB) on hepatitis B virus (HBV) in HBV replication mice. HBV replication-competent plasmid was transferred into male BALB/c mice by using hydrodynamics-based in vivo transfection procedure to develop HBV replication mouse model. The model mice were matched by body weigh, age and serum levels of hepatitis B e antigen (HBeAg) and were divided into four groups:AM group, DDB group, AM+DDB group and NS group, with the last one as control, and the mice of each group were administered corresponding agent orally twice a day, in a medication course lasting 3 d. On the third day, the mice were sacrificed 4-6 h after the last oral intake. HBV DNA replication intermediates in liver were analyzed by Southern blot hybridization. The serum hepatitis B surface antigen (HBsAg) and HBeAg were detected by enzyme linked immunosorbent assay (ELISA). Compared to the animals in the control group, HBV DNA replication intermediates in liver and HBsAg and HBeAg in serum from the AM and AM plus DDB group of mice decreased, and there was no difference between these two groups of mice. The levels of HBV DNA intermediate from liver and the serum HBsAg and HBeAg between the control and DDB group, however, were not obviously different. In conclusion, the inhibition effect of AM on HBV was detected, but treatment with DDB for 3 days did not influence the viral replication and expression of HBV in the HBV replication mice.