Objective To construct heparin-binding epidermal growth factorlike growth factor((HB-EGF)) transgenic mice in order to investigate the role of HB-EGF in the fibrosis of tissues.Method HB-EGF gene was cloned form mouse with RT-PCR.The cloned HB-EGF gene was then inserted downstream of Chicken β-actin promoter to construct the vector which could express HB-EGF gene in system.The transgenic mice were produced by microinjection method and the genotype was detected by PCR with specific primers.The expression profiles of HB-EGF in body tissues were illustrated by Western Blot.Massion staining of liver,lung,kidney and bladder from transgenic mice and nontransgenic control mice was carried out.Results Two lines of HB-EGF transgenic mice were established.The expression levels of the HB-EGF gene in liver,lung,kidney and bladder from transgenic mice were obviously higher than those form control mice.The extents of fibrosis in the liver,lung,kidney and bladder of transgenic mice were significantly enhanced by the expression of the transgene.Conclusion Two lines of system-expressing HB-EGF transgenic mice were established successfully.The over-expression of HB-EGF can significantly aggravate tissues fibrosis degree.
Objective To purify the immunoglobulin from Ricefield eel,and to prepare the rabbit polyclonal antisera.Methods Ricefield eel immunoglobulin was purified using staphylococcal protein A affinity chromatography. The products were examined by SDS-PAGE electrophoresis.Rabbit antisera were prepared by repeatedly immunizing rabbits with purified IgM.And anti-Ricefield eel IgM antibodies in rabbit serum were detected by agarose double immunodiffusion.Western blotting was performed to assess specificity of rabbit antisera for Ricefield eel IgM.Results Immunoglobulin purified from Ricefield eel had only two bands in SDS-PAGE.The titer of rabbit antisera obtained was up to 1∶32 examined by agarose double immunodiffusion.The specific activity of polyclonal rabbit antisera against immunoglobulin of Ricefield eel was indicated with western blotting.Conclusions Ricefield eel immunoglobulin was successfully purified,and rabbit antisera was prepared.This provide a tool for developing the serological test system of Ricefield eel.
Objective To establish a route for administration of drugs to liver directly and reduce their influence on other organs.Methods A catheter was inserted in proper hepatic artery and remained one end of it outside under surgical operation microscope. Results The total number of rats in experiment was fifteen. Ten of them were alive and recovered from operation quickly. The reasons of death were vaseular injury, liver damage, overdose of heparin and accident in anesthesia, which could be avoided by skilled operaters. Conclusion The above-mentioned method is simple, and easy to be mastered when drugs are given through the free end of the catheter.
Familial hyportrophic cardiomyopathy (FHCM) is a compensatory response to defects in sarcomeric structural protein and mitochondrial related proteins. The frequency of this genetic disease is about 0.2%-0.5% and there are up to 6 millions potential patients in China. So far, 10 sarcomeric structural protein genes of β-myosin heavy chain, myosin binding cardiac troponin T protein-C etc and 3 mitochondrial related proteins genes of mitochondrial DNA, AMP-activated protein kinease, γ2, frataxin were found to be involved in FHCM. We summarized the molecular genetic mechanism in the review paper.
Objective To observe the relationship between the concentration of Walker-256 ascites inoculated into the abdominal cavity and immunological function of rats.Methods 36 Wistar rats were randomly divided into 3 groups,12 rats each.In group Ⅰ,the rats were inoculated with 0.3 ml ascites without dilution.The rats in group Ⅱ received 0.3 ml ascites in 1:5 dilution with saline.The rats of control group received intraepritoneal injection of 0.3 ml normal saline only.Immunological function was assessed at 8 days after ascites inoculation.Results In group Ⅰ,apparent ascites was formed only in 2/12 rats,while 11/12 rats in group Ⅱ showed ascites formation.The immune function of group Ⅰ was significantly higher than that of group Ⅱ,and that of group Ⅱ was poorest among the 3 groups.Conclusion Our finding imdicate that very high concentration of ascites perhaps elevate the immunological function of rat.Attention should be paid to select an appropriate concentration of inoculated cancer ascites to avoid drastic changes in the immunological function in rats.
Objective To explore the establishment and postoperative mangement of 2K2C(two kidneys two clips) renovascular hypertensive model of rats.Methods 150 male Sprague-Dawley rats weighing 80 to 120 g were selected to make the 2K2C renovascular hypertensive model.Within 10 weeks after the operation of renal arterial constriction,the postoperative complication,the mortality,the blood pressure were recorded and the spontaneous strokes were confirmed by brain slides examination. Results After the renal arterial constriction,all rats developed hypertension one month later and celiac hemorrhage and infection were the most common complications of the operation. The causes of deaths included postoperative celiac hemorrhage(two cases),hyperanaesthesia(3 cases),incurable severe infection(3 cases),renal failure(5 cases), severe malnutrition unknown causes(2 causes). Brain slides examination was made in two spontaneous hypertensive rats, with basal ganglia hemorrhages confirmed. Conclusion Proficient surgical technique is the key factor in making the 2K2C renovascular hypertensive model of rats and the intensive postoperative manegement can decrease the mortality.
Three kinds of laboratory model mice in human chronic myelogenous leukemia are available, including xenotransplantation of CML cells into immunodeficient mice, bcr/abl retroviral bone marrow transduction and transplantation,bcr/abl transgenic mice. Recent studies in these different model systems have yielded important advances in our knowledge of the pathogenesis of human chronic myelogenous leukemia.
Objective To establish a mouse aplastic anemia model by using cyclophosphamide and methylbenzene, and research its pathomorphology. Method Cyclophosphamide 50.0mg/kg was injected each other day, four times,with inhalation of toluene (30mg/L, 2h/d for 8 days) to reproduce aplastic anemia model. Control group was injected with equivalent saline subcutaneously without narcotics. Result The peripheral hemogram of mice in the model groups showed that there was a remarkable decrease in three types of cells (p0.01) and in bone marrow hyperplasia. And there was a distinct increase in lipocytes. The spleen was atrophied, and its lymphnodule became small or disappeared. The microcirculation of ear was obstructed. All indices conformed to aplastic anemia. After 35 days, the pathological changes still existed. The changes were similar to human aplastic anemia, while there wasn't such changes in the control group. Conclusion The method to reproduce aplastic anemia model was simple, and had a high success rate. It could be used to screen relative drugs.
The major histocompatibility complex (MHC) associates not only with the transplantation rejection reactions,but also with immunological responses in a variety of pathological processes. In this review we presented recent advances in research on the effects of MHCⅠand MHCⅡ molecules in transplantation (interactions with NK cells and T cell), antigenic peptides, gene transfer induced immunological resistance and gene therapy, etc.
Pathogenic Mycoplasmas have been found in humans, animals, plants and insects,which can cause many diseases. However, the diagnosis of Mycoplasmas with culture and serological methods is limited, due to slow growth, extensive nutritional requirements and cross antigens of the pathogens. In recent years, with the progress of molecular biology, polymerase chain reaction (PCR) has been adapted widely for the diagnosis of Mycoplasma infections. This review mainly summarizes the successful application of PCR techniques, such as standard PCR, PCR ELISA, nested PCR, real time quantitative PCR, RT PCR and multiplex PCR to the diagnosis of Mycoplasmas . These techniques provide timesaving, sensitive, specialized methods in Mycoplasmas detection.
Objective To study the catheterization and angiography of hepatic artery via carotid artery route in rats. Methods Cervical and abdominal incisions were made on 60 male Sprague Dawley rats. At the open surgery a catheter was inserted into the hepatic artery via left common carotid artery, and then a hepatic artery angiography was performed. Results The success rates of catheterization and angiography were 95% and 100%, respectively. Conclusion The catheterization and angiography of hepatic artery via carotid artery route in rats is a feasible approach with a rather high success rate of technique and a popular future.
Objective To investigate the effect of Cortex Eucommiae on the fractures healing process. Methods Tibia fracture models were created in 45 rabbits. The animals were divided into experiment (n=23) and control groups (n=22). The experimental rabbits were intragastrically administered with Cortex Eucommiae and the control animals received the same amount of physiological saline. The fracture ends were taken for light microscopic examination on days 7, 17, 27, 37 and 47 of the experiment and the histopathological data were semi quantitatively analyzed. Results The light microscopic observation demonstrated that the repairing responses occurred earlier in the experimental group than that in control group. Conclusion The concentrated extracts of Cortex Eucommiae taken orally accelerates the healing process of fracture in rabbits.
Objective To study the effects of polychlorinated biphenyl(PCB) on bcl 2 and TGFβ1 expression in rat kidneys. Methods 40 male Wistar rats were divided into a control (group Ⅰ) and three experimental groups (fed by 10 -8 mol·L -1 of PCB, group Ⅱ, 10 -7 mol·L -1 , group Ⅲ, and 10 -6 mol·L -1 , group Ⅳ)to establish animal model. Three months later, all rats were killed and the kidneys were taken for routine histopsthological examination and immunohistochemical assay of pcl 2 and TGFβ1 expression. Results The extent of structural damage of the kidneys was related to the doses of PCB, showing more seriously injuries in the high dose group than those of other groups( P 0 05). The PCB induced bcl 2 and TGFβ1 expression was increased in a dose dependent manner. Conclusion PCB can cause structural and functional injuries in rat kidneys.
Objective To evaluate the reliability of enzyme immunoassay (EIA) of HSV-1 as antigen for detection of B virus antibodies. Methods B virus antibodies were detected with enzyme immunoassay (EIA,HSV-1antigen),enzyme-linked immunosorbent assay (ELISA, HSV-1antigen), immunobinding assay (DIA ,B virus antigen), immunofluorecence assay (IFA, B virus antigen) and enzyme-linked immunosorbent assay (ELISA, B virus antigen) in macaque monkey sera. The testing results were compared and analysed. Results Consistency of testing results of EIA(HSV-1antigen)with DIA( B virus antigen), IFA (B virus antigen) and ELISA (B virus antigen)were 100%,95.6% and 97.4%, respectively. The negative sera which were confirmed by EIA (HSV-1antigen) were still negative by IFA( B virus antigen). Among 779 negative sera confirmed by EIA (HSV-1antigen), 2.6%(20) was positive by ELISA (B virus antigen). Conclusion There is much consistency of testing results of EIA(HSV-1antigen)with DIA(B virus antigen), IFA(B virus antigen)and ELISA(B virus antigen). Meanwhile, among B virus free animals which were confirmed using HSV-1antigen, very few animals would be positive using B virus antigen.
Degenerative retinal disease in laboratory and domesticated animals represents a wealth of different pathological manifestations which are important for understanding the basis for human disease. Animals with degenerative retinal disease, homologous to that found in humans, make useful models for investigation of treatment by retinal transplantation, pharmaceutical and gene therapy.
Objective To investigate the expression of heat shock protein (HSP) 27, 70 and 90 in Microtus fortis ovarian cancer. Methods Eight cases of spontaneous ovarian cancer and normal ovary were examined by histopathology and assessed for the expression of HSP27, HSP70 and HSP90 using immunohistochemical SP technique. Results All the 8 tumors were histopathologically diagnosed as serous cystadenocarcinoma. The positive-staining rates of HSP27, HSP70 and HSP90 in the cancers were 100%, 100% and 62%, respectively, while all were negative in normal ovary tissue. Conclusion The results demonstrate that HSP27 and HSP70 are overexpressed in the spontaneous ovarian cancers of Microtus Fortis, and the expression of HSP90 is lower. It is worthy of further study whether an animal model of ovarian cancer can be established from such tumor-bearing Microtus Fortis and to serve experimental studies on ovarian cancers.
Objective To assess the effects of iodine excess on reproduction and memory ability in rats. Methods 60 adult female Wistar rats were randomly divided into three groups: group Ⅰ(control) fed with normal content of iodine, group Ⅱ fed with iodine excess diet and group Ⅲ fed with iodine deficient diet. After 12 weeks, the rats were mated with normal male rats in a ratio of 2∶1, and were observed for their reproduction ability. Thirty 4 weeks filial rats born from rats in each of the 3 groups, were randomly chosen and included in the memory ability experiment measured by electric surge test. Results The pregnancy rate was 85% in the control group, 65% in iodine excess group and 55% in iodine deficient group. The incidence of errors and latency period measured by electric surge test in iodine excess and iodine deficiency groups were significantly higher than those in the normal group. Conclusion Iodine excess may interfere with the reproduction and memory ability in rats.
Diabetes mellitus is a common genetic glucose metabolic and endocrinal disturbance caused by insulin deficiency absolutely or relatively. In the last half century, diabetes has the increasing rates of morbidity and mortality, and has become the third common, frequently occurring and chronic noninfectious disease after cardiovascular disease and cancer. Along with the research and production of diabetes drugs, the animal models are well developed.
Objective To study the changes of nitric oxide(NO),inducible nitric oxide synthases(iNOS) and apoptosis of Kupffer cells,and their relationship following ischemia and reperfusion in hepatocarcinoma and normal liver tissues.Methods The hepatocarcinoma animal models were established by ultrasonography-guided implantation of VX2 tumor tissue pieces into the left-middle lobe of liver of rabbits.The animals were subjected to 60 minutes clamp-induced ischemia of hepatic artery distributing in the left-middle lobe followed by the reperfusion at 0 min,1 h,1 d,3 d and 7 d,respectively.Taking normal hepatic tissues and cancer tissues,the concentrations of NO and iNOS were determined and apoptotic changes were examined.Results The concentrations of NO and iNOS in normal liver tissues were low and kept until reperfusion for 7 days(44.41±2.0,3.70±0.2).In hepatocarcinoma tissues,except of decrease of concentrations of NO,the concentrations of iNOS were significantly increased following reperfusion 0 min to 7 days(18.45±1.9).Apoptotic Kupffer cells in normal liver tissues and hepatocarcinoma tissues were increased and reached a maximum at 1 d and 1 h,respectively(13.69±3.1,13.66±1.8).Conclusion Kupffer cells may play an important role in production of iNOS and NO,and injury of the hepatocarcinoma tissues following the ischemia and reperfusion.
Objective To evaluate the optimal anesthesia in animal experiments. Methods Several animal species ware used in this study: mice, rats, guinea pigs, rabbits, dogs, cats, pigs and gouts. Combined anesthesia was tested on several animal species, in different ways and with different anesthetics, and the results were evaluated. Results The combination of ketamine hydrocholoride, sumianxin and diazepam in a ratio of 4:2:1 was found to be an optimal intravenous anesthetic agent. For canine, feline and swine, combined anesthesia of both intravenous and intramuscular was most satisfactory. Combined use of atropine, which is an anticholinergic agent, can effectively avoid overanesthesia. Conclusion Satisfactory results of anesthesia in experimental animals can be obtained with an appropriate combined anesthesia.
