Abstract:Objective The aim of this study was to observe pregestational ethinyl estradiol (EE)-exposure-induced glucose metabolism alterations and hepatic glucose-metabolism-related gene expression changes in the offsprings of SD rats. Methods Fifty-two female and 52 male SD rats were used in this study. The female rats were gavaged with sesame oil, 50 μg/kg, 200 μg/kg and 800 μg/kg EE for 15 consecutive days. After the end of exposure period, the female rats were mated with male rats and gave birth to next generation. The blood glucose and insulin tolerance in the offsprings were measured on postnatal days 23 (P23) and 25 (P25). The expressions of hepatic glucose-metabolism-related genes were measured by RT-PCR. Results In the female offsprings, the 200 μg/kg EE group had significantly lower fasting blood glucose level than that in the control group (P < 0.05). At 15 min after glucose administration, the blood glucose level in the 800 μg/kg EE group was much higher than that in the control group, 50 μg/kg and 200 μg/kg EE groups (P < 0.01, P < 0.01, P < 0.01). At the time point of 2 h, the blood glucose level of the 50 μg/kg and 800 μg/kg EE groups were both significantly lower than that in the control group (P < 0.05, P < 0.01). The female offsprings in the 50 μg/kg and 200 μg/kg EE groups had significantly higher glucose level after insulin administration than that in the control group (P < 0.001, P < 0.01). In the male offsprings, the 800 μg/kg EE group had a significantly higher blood glucose level than the control group at 15 min after glucose administration (P < 0.01), and the 200 μg/kg EE group had a lower blood glucose level than the control at 30 min after glucose administration. In the male offsprings, the blood glucose level of 50 μg/kg and 200 μg/kg EE groups were much higher than that of the control group (P < 0.01, P < 0.01). In the female offsprings, Glut2 and Lpk mRNA expressions in the 50 μg/kg, 200 μg/kg and 800 μg/kg EE groups were much lower than that in the control group (P < 0.01, P < 0.05, P < 0.05). Gys2 mRNA expressions in the 50 μg/kg and 200 μg/kg EE groups were much lower than that in the control group (P < 0.01, P < 0.01). In the male offsprings, the 200 μg/kg EE group had much higher G6pase and Pepck mRNA expression than in the control group (P < 0.01, P < 0.01).The Glut2 mRNA expression in the 50 μg/kg EE group was much lower than that in the control group (P < 0.01). The Gys2 mRNA expression in the 800 μg/kg EE group was significantly higher than that in the control group (P < 0.01). Conclusions Pregestational EE exposure can lead to impaired glucose tolerance and insulin resistance in female offspring and alterations of key hepatic glucose-metabolism-related gene expression, and these effects are sex-specific,and female offspring is more sensitive to pregestational EE exposure.

Abstract:Objective To investigate the identification and optimal breeding method of caveolin-1 knockout mice, and provide an ideal animal model for further study of the role of caveolin-1 in cerebral ischemic injury and repair. Methods The introduced caveolin-1 gene knockout mice were reared in the SPF laboratory and genomic DNA was extracted from mouse tail tissue by the method of boiling lysis. According to the primer sequences provided by the Jackson Laboratory of America for polymerase chain reaction (PCR) to detect the genotypes, with the four different ways of mating:caveolin-1^+/-heterozygote intercrossing, heterozygous and homozygous caveolin-1^-/- hybrid (orthogonal and pay) as well as homozygous intercrossing. The pregnancy rate, shape characteristics of the filial generation mice and homozygous rate of the parental mice were observed. Results Agarose gel electrophoresis results indicated that the size of molecular weight of the PCR products was about 200 bp and 661 bp, which were consistent with the expected target gene fragment, and identified caveolin-1 gene knockout mice of different genotypes successfully. The results of different mating patterns are basically in agreement with Mendel rule, and the female and male aveolin-1^-/- homozygous mice had a certain ability to reproduce, three different genotypes of mice had no significant differences between the shape features. Conclusions PCR can fast and reliably identify the genotypes of caveolin-1 knockout mice using genomic DNA through the method of boiling lysis. Combining the breeding methods of intercrossing of caveolin-1 heterozygous mice and intercrossing of caveolin-1 homozygous mice may be a good way to obtain enough homozygous mice and homologous wild type mice in a short period.

Abstract:Objective To study the effect of a Chinese medicine, Huanglian Jiedu Decoction (HLJDD), on atherosclerotic plaque, inflammatory factors and regulatory T cells in ApoE^-/- mice. Methods High fat diet was given to ApoE^-/- mice to establish an atherosclerosis model. 40 male ApoE^-/- mice were randomly divided into five groups:model group, simvastatin group, low, moderate and high dose HLJDD groups (n=8). HLJDD was intragastrically administered in a dose of 3.5, 7.0, or 14.0 g/(kg·bw) once dailg for 16 weeks. The dose of simvastatin was 5.0 g/(kg·bw). Another 8 male C57BL/6J mice were taken as control group. At the end of the 29-week experiment, all of the mice were sacrificed. The aortic plaques, level of blood lipids, inflammatory factors, Tregs number and the level of Foxp3 mRNA were detected and analyzed by ELISA, flow cytometry and RT-PCR, respectively. Results Compared with the control group, the aortic plaques were much larger in the model group, and the levels of TC, TG, LDL-C, IL-6, hs-CRP, and TNF-α were significantly higher than those in the control group (P < 0.01 for all). Meanwhile, the levels of HDL-C, IL-10, TGF-β and Foxp3 mRNA were much lower than those in the control group (P < 0.01 for all), and the Tregs numbers were less than that in the control group (P < 0.01). HLJDD regulated the blood lipids in ApoE^-/- mice and decreased the levels of IL-6, hs-CRP, andTNF-α, however increased the levels of IL-10, TGF-β and Foxp3 mRNA. At the same time, it increased Tregs number in the ApoE^-/- mice. Compared with the model group, the difference was statistically significancet (P < 0.01). Conclusions HLJDD can significantly alleviate the aortic plaque damages in ApoE^-/- mice.It may be related to the up-regulation of Tregs, which can lead to decrease the expression of serum pro-inflammatory factors such like IL-10, hs-CRP and TNF-α.

Abstract:Objective To establish a new orthotopic implantation model of hepatocellular carcinoma in BALB/c nude mice. Methods The nude mice were under general anesthesia with isoflurane inhalation. Hepatocellular carcinoma was induced by subcutaneous inoculation of human hepatocellular carcinoma Bel-7402 cells in nude mice. The orthotopic implantation model of hepatocellular carcinoma in nude mice was established by orthotopic implantation of small pieces of the liver cancer tissues into the mouse liver by either a sandwich method or embedding method. The post-operative tumor growth was determined by a small animal in vivo imaging system. Histopathological examination of the liver and tumor tissues was performed at four weeks after modelling. Results The orthotopic hepatocellular carcinoma model of nude mice established by the sandwich method had several distinct advantages:shorter operative time, less operational difficulties, with a high rate of tumor formation, no wide-spread peritoneal tumor dissemination, well-preserved tumor histological structure, and low animal mortality. Conclusions Compared with the embedding method, the sandwich method can save much time for preparation of this orthotopic implantation model of hepatocellular carcinoma in nude mice, and provide a more convenient platform for research of liver cancer pharmacodynamics and other biological studies.

Abstract:Objective To explore a new method of reducing the mortality of rat models of LiCl-pilocarpine-induced status epilepticus (SE) by oral gavage of rehydration salts (ORS) liquid.Methods Forty-five young SD rats (20-day-old) and 30 adult SD rats (60-day-old) were enrolled in this study. They were randomly divided into five groups:the adult SE rats received ORS gavage (adult SE+ORS),adult SE rats(adult SE) and young SE rats received ORS gavage (young SE+ORS), young SE rats (young SE) and young blood glucose group. Seizures were induced in both young and adult SE groups with i.p. LiCl-pilocarpine was injected i.p. to induce SE,which was interrupted by diazepam at 60 minutes after SE onset. ORS gavage was conducted in all the ORS groups in different doses according to their ages at 1 hour, 12 hours or 24 hours after the onset of SE. The other groups were not given any treatment. The latency of seizure onset, changes in body weight, level of blood glucose at 8 hours after SE onset and finally the death rate of the rats were recorded. Results (1) The body weight of the rats with seizures was reduced by 5%-8%.(2)The mortality rate of the SE groups was high, especially at 24 hours after the SE onset, which was 40% in the young group and 57% in the adult group. (3) The blood glucose level was significantly decreased in the SE rats, and ORS gavage made the blood glucose returning to normal in the SE rats. (4) ORS gavage apparently decreased the mortality rate of young and adult groups during the 72 hours after seizure onset. Conclusions ORS gavage effectively reduces the mortality of SE rats, and is an effective method to improve the survival of SE rats.

Abstract:Objective The aim of this study was to observe and analyze the differences of reproductive system in fertile and infertile male naked mole rats. Methods Five fertile and 5 infertile 2-5-year old male naked mole rats were used in this study. The testis and epididymis on one side were collected, weighted, and measured to get their viscera coefficient. The other side testicle was collected and fixed in neutral formalin solution for histological examination.The other side epididymis was used to examine the sperm count and sperm activity. Serum of the naked mole rats was collected to measure the level of luteinizing hormone (LH) and testosterone (T). Results Compared with the fertile group, the testis weight of infertile group was significantly decreased (P < 0.05), their epididymis weight, testicular coefficient and the epididymis coefficient were all significantly decreased (P < 0.01), all their sperm count and sperm activity and their LH and T levels were significantly decreased (P < 0.01 for all). Pathological examination of the testicular tissues of the fertile group showed that many types of spermatogenic cells were seriously detached, the numbers of Sertoli cells and stromal cells were decreased. Mature sperms in the seminiferous tubules were highly decreased in number or even completely disappeared. Conclusions Compared with the fertile individuals, the reproductive system of infertile male naked mole rats presents structural atrophy and declined function.

Abstract:Objective To investigate the effect of chronic corticosterone administration on learning and memory and expression of synapse-related proteins in mice. Methods C57BL/6J mice were randomly divided into control and corticosterone (CORT) groups (10 mice per group). CORT was dissolved in the vehicle (0.45% hydroxypropy-β-cyclodextrin, β-CD). Corticosterone (5 mg/kg/day) was orally administered in drinking water for 35 days. The control mice received vehicle (β-CD) in drinking water during the entire experiment. Social discrimination test and Morris water test were applied to evaluate the learning and memory impairment of the model mice. Results Both the mice of control and CORT groups were more interested in social (stranger cage) than the non-social (empty cage) experience. The control group increased the time spent with an unfamiliar mouse (stranger 2 cage). The mice of CORT group were not able to discriminate between an unfamiliar (stranger 2) and the familiar mouse (stranger 1). In the Morris water maze test, in contrast to the first 3 days, an increase in latency to reach the hidden platform (P < 0.05) was observed on day 4 in the CORT group compared with the mice of control group. CORT-treated mice showed a statistically significant decrease in the frequency in the platform, frequency in the target quadrant and distance in the target quadrant (P < 0.05, P < 0.05, P < 0.05) compared with the control group. The western blot analysis showed a notably decreased expression of PSD95, Syn-1 and p-erk in hippocampus in the mice of CORT group compared with the control group. Conclusions Chronic administration of corticosterone induces learning and memory impairment and decreases the expression of synapse-related proteins.

Abstract:Objective To establish a stable mouse model of limb heterotopic composite tissue allograft transplantation. Methods 25 pairs of C57BL/6 mice were used for this limb heterotopic allograft transplantation experiment. The hind legs were taken from the donor mice. A transverse incision was made at the level of inguinal ligament to expose and mobilize the femoral vessels up to the external iliac vessels. All branches to the external iliac artery and vein were cut off by electrocoagulation. To cut off the external iliac artery and external iliac vein, and to transect the femur, thigh muscles and skin. The donor external iliac artery-external iliac vein anastomosis was performed in end-to-side fashion with recipient mice's right common carotid artery and jugular vein, respectively. Continuously suture the donor skin to the recipient skin incisal margin to fix the limb. Cut off the donor limb's distal tibial joint, and ligate the stump. Results The success rate of the heterotopic limb transplantation operation was 76%. The total operation time was 87.1±10.1 minutes, and the vessel anastomosis time was 55.3±9.6 minutes. Conclusions This mouse model of limb heterotopic limbcomposite tissue allograft transplantation shows a high success rate of the surgical operation, and is stable and reliable. It is a new ideal model for studies on transplantation immunology.

Abstract:Objective To observe the effects of tacrolimus on blood glucose, insulin, expressions of protein phosphatase 2A and P-AKT in rats in order to explore the mechanism of hyperglycemic action of tacrolimus. Methods Sixty male SD rats (body weight 89.83±4.44 g) were randomly divided into tacrolimus group (n=40) and control group (n=20). The rats in the tacrolimus group were administrated with tacrolimus 4 mg/kg daily. The rats in the control group were given the same amount of normal drinking water daily. The rat body weight, fasting blood glucose concentration and blood concentration of tacrolimus were measured monthly. All rats were killed at 5 months after the tacrolimus administration. The serum insulin levels were detected by radioimmunoassay method. The expressions of PP2A and P-AKT in liver tissues were assessed by immunohistochemistry. Results After two months of administration, the blood glucose levels in the tacrolimus group were significantly higher than those in the control group. The HOMA-IR in tacrolimus group was significantly higher than that in the control group P < 0.05). ISI was significantly lower than that in the control group (P < 0.05). Immunohistochemical examination showed that the expression of PP2A in hepatocytes in the tacrolimus group was increased compared with the control group, while expression of P-AKT in hepatocytes of the tacrolimus group was decreased than that in the control group. Conclusions Tacrolimus can induce necrosis of islet cells, decrease of the amount of islet cells and insulin secretion, decease of sensitivity to insulin, and increase the resistance to insulin, therefore, leading to increase the blood glucose level in rats. The expression of PP2A in hepatocytes in the tacrolimus group is increased, while the expression of P-AKT is decreased. Interfering of insulin signal transduction pathways may be involved in the hyperglycemic effects of tacrolimus.

Abstract:Objective To construct the eukaryotic expression vector pEGFP-N1/IL-37b and analyze the expression of IL-37 gene in THP-1 cells. Methods Total RNA was extracted from human peripheral blood mononuclear cells (PBMCs) and the coding region of IL-37b gene was amplified by RT-qPCR. Then, the gene was cloned into pEGFP-N1 eukaryotic expression vector. After transfected the recombinant plasmid into THP-1 cells, the expression of IL-37 was detected by RT-qPCR and Western blot. Results Double restriction enzyme digestion and gene sequencing showed that IL-37b gene was correctly inserted into the eukaryotic expression vector pEGFP-N1.RT-qPCR and Western blot showed that the IL-37 expression level was increased significantly (P < 0.01) after transfection in THP-1 cells. Conclusions We successfully constructed a novel anti-inflammatory cytokine IL-37 eukaryotic expression vector pEGFP-N1/IL-37b, which lays a foundation for further study on IL-37 functions and its association with related diseases.

Abstract:Objective To investigate the differential expression of integrin alpha v beta 3(αv,β3) on bovine endometrial epithelial cells before and after co-culture with bovine blastocysts, and to explore whether this specific signal might be applied as a new marker for identifying the bovine uterine receptivity. Methods The in vivo bovine embryos were co-cultured with their endometrial epithelial cells for 24 h, then, RT-PCR was used to detect the differential expression of αv,β3 among those groups. Result The results showed that αv,β3 can be expressed on bovine endometrial epithelial cells both before and after co-culture with their in vivo embryos. There were no significant differences of expression of αv,β3 between Group I (only bovine blastocyst) and the control one (P > 0.05), but there were significant differences of αv,β3 among Group Ⅱ (the hatched bovine blastocyst), and Group I, the control one (P< 0.05). Conclusions The in vivo hatched bovine blastocysts are more suitable for induction of intergrin αv,β3 in bovine endometrial epithelial cells after their co-culture than that of co-cultured with early stage blastocyst. Integrin αv,β3 might be applied as a new molecular marker for detecting bovine uterine receptive status of the bovine endometrium.

Abstract:Objective To study the effect of Maca extract on toxicity and immune organs in mice. Methods One hundred and fifty SPF BALB/c mice (male:female=1:1, body weight 20-22 g) were used in this study. The mice received Maca extract for 15 days, and the maca toxic effect on mice, and the thymus and spleen weights were observed. Results At 24 hours after oral administration of Maca extract in an accumulated dose of 320 g/kg·bw, no toxic response was seen in the mice. After administration of Maca extract in a dose of 16.0, 32.0, and 64.0 g/kg·bw for 15 days, the thymus weight was increased by 36.84%, 89.47% and 107.89%, respectively, the spleen weight was increased by 44.83%,62.01%, and 89.66%, respectively, the carbon clearance index was increased by 30.85%, 30.85% and 42.55%, respectively, and the serum hemolysin value was increased by 11.64%, 20.03% and 31.51%, respectively, in the experimental mice, significantly higher than those of the control group (P < 0.05 or P < 0.01 for all). Conclusions Maca extract exerts an enhancing effect on the immune organ weights and improve the immune function, and is associated with a lower toxicity.

Abstract:Objective To observe the expression of uPA, tPA and PAI-1 in whole blood of rat membranous nephropathy (MN) models induced by cationic bovine serum albumin (C-BSA), and to explore the effect of fibrinolytic system on podocyte apoptosis and pathological changes. To explore the possible preventive and therapeutic effects and the possible mechanisms of early prevention of fibrinolysis. Methods We developed a MN model with the modified Border method. At the end of the 1st, 2nd, 3th, and 4th week of immunization, respectively, the levels of whole blood uPA, tPA and PAI-1 were determined by ELISA. The rat kidney tissues were examined by light microscopy and electron microscopy to identify the pathological changes. The expression levels of nephrin and WTl were detected with immumofluorescence staining and their correlation was analyzed. Results Compared the treatment group with control group, the levels of whole blood uPA, tPA and PAI-1 of the model group were decreased, while PAI-1 was elevated, showing a significant difference (P < 0.05). The degree of renal interstitial fibrosis was more serious. Correlation analysis showed that the whole blood tPA and uPA levels were positively correlated with the changes of nephrin protein expression in the kidney tissue, while the whole blood PAI-1 level was negatively correlated with the nephrin protein expression in the kidney tissue. Conclusions In the process of MN development, the fibrinolytic system may have important significance for podocyte apoptosis. Determination of early phase of MN podocyte injury may be another therapy target for prevention of the disease development, and then provide new ideas for clinical research and drug development for MN.

Abstract:Objective To describe the phenotype of NK cells in Bama miniature pigs, and establish an efficient activation and culture method for porcine cytokine-induced killer (CIK) cells in vitro. Methods The porcine peripheral blood mononuclear cells (PBMCs) were isolated by Percoll gradient centrifugation, and the phenotype of NK cells was tested by detecting the CD2⁺/CD8⁺/CD3^- cell compartment. To establish an efficient activation and culture method for porcine CIK cells, we optimized the culture conditions to improve the CIK activation efficiency. Results Using the optimized induction culture conditions, the ratio of CIK (CD2⁺/CD8⁺/CD3^-) cells was up to 43.63% at the fifth day, approximately 5.59 times increased compared with the initially separated PBMCs. Cell proliferation experiments showed that three obvious fluorescence peaks were observed on the fifth day. The results indicated that the induced CIK cells underwent three times cell division, in theory, about increased 8-fold compared with the initial separation of PBMCs. Furthermore, the qRT-PCR result of the surface markers of porcine NK cells also showed a similar variation tendency as the flow cytometry results. Conclusions Our findings demonstrate the successful establishment of an efficient activation and culture method for porcine CIK cells in vitro.

Abstract:Objective To establish a simple and sensitive detection method of Sendai virus (SeV) by reverse transcription loop-mediated isothermal amplification (RT-LAMP) technique.Methods According to the published GenBank sequences (DQ219803.1), six pairs of primers were designed targeting the conserved region of SeV. The amplification products were detected with a LAMP real-time Turbidimeter.(LA-302). Through optimizing the LAMP primers and reaction conditions, a rapid and specific detection method of SeV was established.Meanwhile, the amplified products were colored by fluorescence detection reagent after completion of the reaction, so that the amplification could be visualized and detected by naked eyes. Then, methodological evaluation of the RT-LAMP was tested. Results The method of RT-LAMP showed a highly efficient amplification for SeV viral target gene which was performed at 63℃ for 60 min with the LAMP real-time Turbidimeter (LA-302).The detection limit was 2.1 TCID₅₀, 100 times higher than that of RT-PCR, and no cross-reaction with other RNA and DNA viruses of mice was observed. The results of SeV LAMP reaction was visualized and the tube could be directly observed by naked eyes with the addition of fluorescence detection reagent. The results were consistent with the results detected by real-time tubidimeter.92 clinical samples were detected byRT-LAMP, RT-PCR and indirect ELISA, and the coincidence rate was 100%.Conclusions This established SeV RT-LAMP detection method is fast, specific, highly sensitive,easy to perform under simple conditions, and is suitable for rapid detection of Sendai viirus.

Abstract:Objective To analyze the DNA genetic diversity in SPF Jinding duck population using microsatellite markers. Methods The DNA genetic diversity in 71 SPF Jinding ducks were analyzed by microsatellite markers. Results A total of 152 alleles were detected at 17 microsatellite loci, and the numbers of alleles at each locus varied from 3 to 13. Thirteen microsatellite loci showed a high level of average PIC values (PIC > 0.5), the hererozygosity (H) was 0.5816±0.0142. Other loci showed a middle level of the average PIC values. Only 5 microsatellite loci were in Hardy-Weinberg equilibrium, although 12 microsatellite loci were not in (P < 0.01). Conclusions The results revealed that the genetic variabilities in the SPF Jinding duck population are rich, and this population meets to the genetic characteristics of the closed colony animals, so it can be used to establish the closed colony of SPF Jinding duck.

Abstract:Objective To determine the interference effect of H.hepaticus infection on the functional characteristics of dendritic cell (DC) surface molecules and immune response in mice. Methods Male BALB/c mice were inoculated with H.hepaticus (ATCC 51450).Murine bone marrow-derived dendritic cells (DC) were isolated and co-cultured which were stimulated by GM-CSF and IL-4 at the fifth month after the last inoculation. Then the DCs were subjected to FACS analysis for surface markers (CD11c, CD40, CD80 and MHCⅡ) detection.On this basis, virus suspension of Newcastle disease virus(NDV) ZJ1 strain was inoculated into the mice. Serum was collected for detection of the NDV antibody titer in serum weekly to explore the difference of antibody titer between the two groups.Results The expression rates of CD40 and MHCⅡ on the mouse DCs in experimental group were higher than that in the control group. The NDV antibody titer of experimental group was slightly lower than that in the control group in the first week. During the 2nd to 5th weeks, the titer was higher than that in the control group, with a very significant difference. In the 6th week, the titer of both the two groups tended to fall.Conclusions H.hepaticus infection can promote bone marrow DC maturation in mice, stimulate the expression rates of MHC Ⅱ and CD40, and enhance the NDV antibody levels.

Abstract:Objective To more intuitively understand the quality control for laboratory animals and further achieving a more scientific and reasonable management of laboratory animals, the infection index as evaluation criteria was introduced. Then the best way to calculate infection index was explored in order to more scientifically reflect the infection status of laboratory animals. Methods Infection index, also called the degree of infection, is a qualitative indicator of monitoring laboratory animal quality. After arranging, analyzing, processing and gathering the data from laboratory animal quality monitoring, the index reflects synthetically the pathogen infection status or trend of a particularly investigated experimental animal population or the development of certain experimental animals. Results In general, the pathogen infection index of mice was slightly decreased, while the pathogen infection index of rats roughly increased year by year. In comparing infection index by different pathogens, the parasite infection index of mice was found to be higher than bacteria and virus infection indexes, while the bacteria infection index of rats was higher than parasite infection index and virus ones. Conclusions The infection index model intuitively reflects the quality control status of laboratory animals. The analysis also reveals that the parasite monitoring of the mice and the bacteria detection of rat needs to be reinforcement. In addition, the index of infection reveals that the pathogen infection of mice is well under control, while that of rats tends to be more serious year by year.

Abstract:Naked mole rats are unique animals with long lifetime, anti-tumor properties, hypoxia tolerance, low metabolic rate, analgesia, tactile sensitiveness, poor eyesight and strong bone regeneration ability. Based on the basic biological characteristics of naked mole rats, and the current research trends in the field of cancer, aging, hypoxia tolerance, as well as algesia, this review focuses on the application prospects of naked mole rats in biomedical research.

Abstract:The incidence of depressive illness is high worldwide, and the inadequacy of currently available drug treatments contributes to the significant health burden associated with depression. Animal models of depression used as the main methods to study the pathogenesis mechanism and select effective drugs receive increasing concerns. Current popular models of depression creatively merge ethologically valid behavioral assays with the latest technological advances in molecular biology. In this context, this study aims to review the animal models of depression and pathogenesis related with face validity, construct validity, and predictive validity of these models. These models include stress-induced models, injury-induced models, drug-induced models and transgenic models which all mimic the depression symptoms of human to some degree and are of great value for developing new antidepressant drugs and studying the pathogenesis of this disease.

Abstract:Diabetes mellitus (DM) complicated with atherosclerosis (AS) is the most common cause of amputation and death in patients with type 2 DM due to the instability of plaque, severe ischemia and the high rate of restenosis after treatment. It is important to reduce the diabetic vascular complications in those patients. Establishment of animal models of severe diabetic vasculopathy will provide an experimental basis for further studies on those complications. In this paper, we will firstly discuss the mechanism of the development of severe diabetic vasculopathy and then review the methods referring to the establishment of this kind of animal models worldwide.