Abstract:Laboratory animals support the life science research. Based on their clear genetic, immune background, mice have been widely used. In the Post-genomics era, a great deal of genetic engineering mouse models, including those transgenic mice and gene targeted mice had been developed to study some a specific gene. However, most human diseases are not controlled by a single gene; besides the environmental factors, multiple genetic factors can influence the development of human diseases. The complex multi-gene traits research in life science put forward new challenges to the laboratory animals. To meet this challenge, scientists have proposed to develop a new mouse resource called the Collaborative Cross (CC) mice. This paper will review the background and character of CC mice, and gives some suggestions for its application.

Abstract:Objective To evalutate the safty of hBMSCs transpalntation and to observe their migration and distribution in the brain of young macaca fascicularis. To establish a new technology platform and theoretical basis for the treatment of central nervous system diseases in children.Methods Labelled hBMSCs were transplanted into the striatum of young macaca fascicularis. Brain sections were examined to evalutate the inflammatory reaction and immunological rejection of local injection sites by HE observation and immunohistochemical staining. Migration and distribution of transplanted-hBMSCs was observed by real-time fluorescence quantitative PCR of male DNA and fluorescence microscope.Results The results showed that the direct intracerebral injection of hBMSCs did not cause systemic symptoms in animals. There is no inflammatory reaction and immunological rejection was detected, and degeneration and necrosis of neural cells and proliferation of glial cells were absent in the local injection sites. The transplanted hBMSCs survived, and migrated into the brain after 4 weeks transplantation. Its migration and distribution have certain regularity and were overlapping between transplant recipients. In addtion, hBMSCs tended to extend rostrally into the forebrain and showed preference of migrating toward the blood vessels and below the ependyma.Conculsions Intracerebral transplantation of hBMSCs is safe. And hBMSCs can survive and migrate into the brain.

Abstract:Objective Study on the blood pressure, renal sodium excretion, related renal sodium ion channels and signaling pathways between male and female mice, to investigate the relationship between gender differences in blood pressure and renal sodium ion channel.Methods Blood pressure of adult female and male mice was measured using tail-cuff sphygmomanometer. Plasma and urine electrolytes were performed using flame photometer.Protein levels of renal sodium transporters NCC, NKCC2, ENaC were determined by western blot, mRNA levels of these transporters and the upstream With No lysine (WNK) kinases were determined by Real-time PCR.Results The blood pressure of female mice was significantly lower than that of male mice. Renal sodium excretion and mRNA levels of NCC&NKCC2 were increased significantly in female mice. No obvious difference was observed in ENaC. The mRNA level of WNK4 was increased while no change in WNK1.Conclusions The lower blood pressure of female mice may caused by increasing renal sodium excretion and WNK4-NCC/NKCC2 signaling pathway involved.

Abstract:Objective Establish a Long-term Survival Rat Model of Sepsis for recovery naturally, and the idea of researching about sepsis survivors may point out new research ideas and methods for sepsis. Methods 40 SD rats were divided into the sham group (8) and the CLP group (32), with all surgical operations under the small animal anesthesia machine and sevoflurane inhalation anesthesia. PVC tubes were inserted into the right carotid artery and jugular vein, respectively. After a postoperative recovery of 24 hours, the rats in the CLP group received the cecal ligation and puncture (CLP), which caused sepsis. The rats were then transferred to the single cages in the recovery room (with the temperature kept between 22~25℃). After the surgery, we commenced fluid resuscitation, consisting of a solution 6% hetastarch and 5% Glucose in the ratio of 1:1. The amount was 20 mL/kg/12h on the first and second days after the surgery. Then it was halved until the rats started eating. These rats were observed about performance, weight and concentration of IL-10 until 30 days after the surgery, when they were anatomized and the change of their organs was watched. Results (1) The survival rate was 75% at 24 h, 62.5% at 72 h, 50% on 7 d after CLP in the CLP group. (2) Based on the severity of sepsis evaluation system, 32 rats at 24 h after CLP reached moderate to severe sepsis. (3) The body weight were decreased in both survival group and the sham group after CLP. The body weight of rats in survival group fell from the fourth day of survival was significantly (P=0.017), and to a minimum after the first six days, compared with the original weight loss (8.51±2.23)%. The body weight of rats in sham group fell to a minimum after the first four days, by which the lowest body weight loss (2.73±1.82)% than the original weight. The difference between two groups was statistically significant (P=0.026). After 30 days, the maximum weight increase rates[sham group (16.16±2.39)% vs survival group (13.03±3.74)%] were not different significantly (P=0.29). (4) Compared the preoperative (0 d) with postoperative 1d, all the plasma concentration of IL-10 in three groups was increased, which in survival group (P=0.000) and in dead group increased significantly (P=0.010). (5) Shown by abdominal anatomy, a large set of foetor and bloody ascites, black bowel, with no encapsulated abscess and adhesion in dead group; Abdominal adhesions, omental with losing shape and gloss and tending to ligation intestine, but no encapsulated abscess in survival group; no abnormal anatomy in sham group. The weight of spleen in total body weight ration was (2.64±0.37) ‰ in survival group and (1.63±0.20) ‰ in sham group. The difference between two group was statistically significant (P=0.032). Conclusions This experiment provided reliable measures of the CLP and methods of screening model, for the establishment of CLP in severe sepsis model of long-term survivors, which is basically in accordance with the occurrence and development of sepsis clinically.

Abstract:Objective Establishing the drug-resistant Candida albicans disseminative infected mice model for new drug screening. Methods The disseminative infected mouse model was generated by intravenously injecting a clinical Drug-resistant Candida albicans strain (CaR) to immunosuppressive ICR mice. The features of model was evaluated by clinical symptom, survival condition, fungal burden in tissue, histopathology, cytokines assay and medication. Results After infected with CaR (0 day), the death of mice started at the first day, though, compared to clinical drug sensitive strain (CaS) infected group, the difference of mortality rate in 16-day observation period was not significant in two groups (CaR, 90.7%; CaS, 86.2%, P=0.158), mice in CaR group died faster than those in CaS group at the early stage; On the fourth day of infection, Candida albicans could be detected in the different tissues, and we found fungal burden in kidney and brain was a significant difference. The typical granuloma caused by fungal infection was the main histopathological feature observed in the kidney, brain and heart. Cytokines in renal tissue were detected by flow cytometry, The changes of IL-1α,IL-6,TNF-α and IFN-γ in kidney were significant. Compared with CaS group, IL-1 and IFN-γ were significantly higher and TNF-α decreased significantly in CaR group.The mice of groups CaR and CaS were treated with 10 mg/kg fluconazole, the mortality rates were 83.3% and 37.5%, which have a significant difference. Conclusions In this study, we successfully established a drug-resistant Candida albicans disseminative infected mice model which is potential tool for the development of new anti-infectious agent.

Abstract:Objective To evaluate the renal toxicity of vancomycin hydrochloride and irbesartan tablets using the zebrafish model. Methods After construction of AB zebrafish kidney model, the fish were treated with drug after fertilization 2 days (2 dpf) to 5 dpf. At the end of the experiment, the number of renal edema zebrafish was counted in each experimental group to evaluate the renal toxicity of drugs. Results The zebrafish development was normal and no obvious toxicity at the dose of 16.4 ng/fish (1/10 MNLD)for vancomycin, and zebrafish renal edema occurred rate was 3.3%, 10% and 10% respectively at the dose of 54.7 ng/fish (1/3 MNLD), 164 ng/fish (MNLD) and 273 ng/fish (LD₁₀) with the death rate of 0%, 0% and 16.7%, respectively, which indicated that there was significant renal toxicity of vancomycin at the dose of 54.7 ng/fish (1/3MNLD) to 273 ng/fish (LD₁₀). Irbesartan didn't induce renal toxicity at the dose of 8.3 μg/mL (1/10 MNLC) to 91 μg/mL (LC₁₀). Conclusions The zebrafish model of renal toxicity can be used for the early evaluation of drug renal toxicity and we made evaluation of the renal toxicity of vancomycin and irbesartan with this model.

Abstract:Objective To investigate biological parameters and morphology in spontaneous type 2 diabetes C57BL/KsJ-db/ db mice and mice with type 2 diabetes mellitus which induced by high-fat diet and streptozotocin injections. Methods C57BL/6J mice were divided randomly into normal control group, Induced Group, and Spontaneous Group with 10 mice in each group; Induced Group: The 11 weeks old C57BL/6J mice were fed on a high-fat diet before intraperitoneal injections of STZ; Spontaneous Group: 8 weeks old C57BL/KsJ-db/db mice (db/db); The body weight, Fasting blood glucose (FBG), oral glucose tolerance test (OGTT) was detected every week for 28 days. FINS, AUC, IR index and pro-inflammation factors (TNF-α, IL-18, IL-1β, INF-γ) were evaluated in the beginning and 8^th week. At the end of experiment, the mice were sacrificed for organ and morphological study. Results The weight of Spontaneous Group were higher than Induced Group, both of them are higher than normal group significantly; Non-fasting blood glucose concentrations of Spontaneous Group mice were higher over Induced Group; In 8^th week the abnormal glucose levels of Spontaneous Group extraordinary stability, however, in Induced Group drops obviously;the FINS was increased in Spontaneous Group, but have no significantly change in Induced Group,both AUC and HOMA-IR index Show a significant downtrend in Induced Group, However, in Spontaneous Group, the rising trend of AUC and HOMA-IR are stabe; The content of the pro-inflammatory factor (TNF-α, IL-18, IL-1β, INF-γ) in both diabetes animal are more than normal group. In 8th week the content of pro-inflammatory factor was decreased in Induced Group, but increased in Spontaneous Group significantly; Pathologic results showed that pancreas, kidney, liver, muscle, and testis, appeared serious inflammatory pathological deterioration in two kinds of diabetes models mice. Conclusions During 8weeks the Spontaneous diabetes animal showed abnormal glucose metabolism, and high concentration of pro-inflammatory cytokines which Symptoms of type 2 diabetes become more and more serious.; glucose metabolism and inflammation showed improved in induced animal model.

Abstract:Objective To establish and evaluate the animal model induced by inhalation injury of airborne fine particulate matter (PM2.5). Methods We manufactured equipment for rats aerosol inhalation with PM2.5. The effects of several facters such as concentrations(100±10 μg/m³、150±10 μg/m³、200±10 μg/m³)、time(1w、2w、4w、8w、12w)、method (non-exposed intratracheal instillation method and aerosol inhalation) and animals (Wistar rats, BN rats and guinea pigs) were investigated to establish the model. The respiratory rate, forced vital capacity ratio of forced expiratory volume (FEV₁/FVC) and arterial partial pressure of oxygen (PO₂) were measured, the pathological changes of bronchial and lung tissues under light microscope were observed. The success animal model was builded as the pneumonia was observed from the pathological changes of lung tissue. Results The Wistar rats exposed to PM2.5 aerosol inhalation for 8 weeks, we can see that the weight growth rate of rat decreased, WBC count and mononuclear cells count increased, the macrophages ratio decreased in BALF, the respiratory rate of lung increased while arterial PO₂ and FEV₁/FVC decreased, inflammation and pulmonary fibrosis changes were observed by bronch and pulmonary pathology, inflammatory changes with a dose-response relationship were observed. Exposed to PM2.5 aerosol inhalation for different time(1 w、2 w、4 w、8 w、12 w)with same dose, the score issue lesions of lung and bronchus in Wistar rats increased and the 8w group is obvious. The Wistar rats exposed to PM2.5 with different method (aerosol inhalation and non-exposed intratracheal instillation method) for 8w, the aerosol inhalation worked as effectively as perfusion while mortality rate of aerosol inhalation is lower. Different animals (Wistar rats, BN rats and guinea pigs) exposed to PM2.5 aerosol inhalation for 8w, the same results were observed with three method respectively while mortality rate of Wistar rats lower. Conclusions The optional conditions that the Wistar rats were continuously inhaled for 8w PM2.5 with a dose of 150±10 μg/m³ were established. The animal model could be used on a national scale, especially in Fujian province. The results would be useful for the development of the research of the prevention and countermeasures of PM2.5 pollution.

Abstract:Objective To explore effect of Xylazine hydrochloride on Bama minipigs under general anesthesia. To emphasize safety consciousness of general anesthesia. To research cardiac main function and structure of normal Bama minipigs in preparation for the subsequent comparative medicine research. Methods 43 Bama minipigs,inject in post aurem muscles of neck with 5 mL of mixed drug conclude Xylazine hydrochloride (2 mL), Atropine Sulfate(1 mL) and Droperidol(2 mL) on each one. Echocardiography after general anesthesia.Observe induction and recovery time of anesthesia, anesthesia maintaining time, total check time and the others. Introduce the method of simple endotracheal intubation.Results Anesthesia, induction period (18±3)min, maintaining period (40±5)min, recovery period (60±10)min. Echocardiography, LAD (2.54±0.20) cm, LVDd (3.41±0.25) cm, LVDs (2.28±0.23) cm, IVSTd (0.60±0.07) cm, LVPWTd (0.59±0.07) cm, AoD (1.77±0.18) cm, EDV (48.59±8.31) cm, ESV (18.28±4.46) mL, SV (39.30±5.16) mL, LVEF (62.76±5.01)%. Conclusions Intramuscular injection of xylazine hydrochloride with droperidol and atropine sulfate on bama minipigs for general anesthesia is a highly conserved specie in cardiovascular system and safe. We obtained some information of cardiac main function and structure of normal Bama minipigs which could provide reference for scientific research and veterinarian clinic.

Abstract:Objective This study was designed to investigate the morphology of vascular calcification in aged rats and the expressions of osteopontin and osteocalcin in the arterial wall of old rats. Methods 20 20-month-old SD rats were recruited in experimental group and 20 3-month-old SD rats were recruited in control group. Aorta, heart and kidney of the rats were collected for assay. HE staining and Von Kossa staining were used to determine the vascular calcification. The expressions of OPN and OC in vascular wall were tested by Immuno-histochemistry and Western-blot. Results In the experimental group, vascular smooth muscle cells are disordered arrangement, thickness of membrane layer is increased, elastic plate fracture and the vascular fibrosis is increased. In the control group, vascular endothelial cells and smooth muscle cells are arranged in neat, elastic plate is complete. In the experimental group, the large artery and small artery were observed the calcium salt deposits. In the control group,arteries were not observed calcium salt deposits. Compared with the control group, the expressions of OPN and OC were significantly increased in the experimental group (P<0.05). Conclusions Vascular medial calcification may be pathological features of vascular aging. Vascular calcification may be related with the up-regulated expression of bone-associated protein OPN and OC.

Abstract:Objective To analyze the electrocardiography (ECG) data of pressure overload-induced chronic heart failure rats.Methods Totally 20 SD rats were randomly divided into sham operation group and heart failure group. Heart failure rats were induced by abdominal aorta constriction.Echocardiogram measurement demonstrated the occurrence of cardiac function.Two lead ECG parameters of limb a was measured and statistically analyzed.Results Ten weeks after operation, there was a increase in heart rate, P amplitude, P duration and R amplitude comparing by those of the sham operation group (P<0.05). ECG showed a significant and ubiquitous J point elevation (P<0.05), with ST segment notable depression (P<0.05).Conclusions ECG in pressure-overload chronicity heart failure rats exhibits obviously characteristic features. ECG is an useful tool for objective and accurate assessment of cardiac function in rats.

Abstract:Objective To explore the effect of liraglutide on pressure-overload chronic heart failure in rats and related mechanisms. Methods Totally 30 SD rats were randomly divided into sham operation group, heart failure group and liraglutide group.The animals were anaesthetized and a Millar pressure volume conductance catheter SPR-838) was inserted through right carotid artery into LV to measure pressure-volume (P-V) loop.Body and organ weight were measured. After the end of intervention, the rats were anesthetized, and abdominal aortic blood taken from the upper serum after centrifugation. Kit method was used to measure superoxide dismutase (SOD), brain natriuretic peptide (BNP), malondialdehyde (MDA).Results Compared with those of the sham operation group, there was a increase in absolute heart weight and (P<0.05). During the P-V loop analyses, We found that left ventricular (LV) end-systolic volume, end-diastolic volume, end-systolic pressure, end-diastolic pressure and maximum pressure were all remarkablely lower in liraglutide group than HF group in rats (P<0.05). Peak rate of pressure rise (dP/dt max), peak rate of pressure decline (-dP/dt min), ejection fraction and Maximum Power were increased in liraglutide group comparable by HF group (P<0.05). SOD activity was significantly increased and BNP concentration was significantly decreased in liraglutide group compared to HF group (P<0.05). Conclusions These results show that liraglutide protects pressure-overload rat heart from failure possibly through reducing oxidation.

Abstract:Objective To establish a rapid monitoring method of the three common bacteria in mice frozen resources, such as embryo, sperm, etc. Methods To extract DNA of the three positive bacteria(Staphylococcus auerus, Klebsiella-pneumoniae and β-hemolyticstreptococcus), and establish PCR monitoring method of the three positive strains through designing primer and refining PCR condition. Then extract total DNA of the frozen resources, detect the DNA according to the PCR condition of the three positive bacteria, some samples were detect by fluorescence quantitative PCR at the same time. Results ①successfully establish a PCR detection method of the three positive bacteria, the minimum detectable concentration of Staphylococcus auerus, Klebsiella pneumoniae and β-hemolytic streptococcus is 4.19×10^-5 ng/μL, 1.98×10^-5 ng/μL and 1.07×10^-3 ng/μL.②Proved that the three bacteria doesn't exist in the sample by normal PCR and fluorescence quantitative PCR methods. Conclusions Establising a rapid monitoring method of common pathogens of frozen embryo and sperm in mice.

Abstract:MiRs display an important role in a variety of biological, physiological and pathological processes, including cell proliferation, differentiation, apoptosis, individual development, occurrence and progress of diseases. Recent studies have discovered that miR-424 is the significant regulatory factor of angiogenesis, and is involved in many diseases such as infectious diseases, vascular diseases, central nervous system diseases and genital system disease. This article reviews the expression, effect and possible mechanisms of miR-424 in non-tumorous diseases.

Abstract:In spite of much progress on its mechanism, diagnosis and treatment, diabetes mellitus remains a public health challenge. The harm of diabetes is not significantly reduced, instead shows an increasing tendency year by year. To achieve an in-depth and comprehensive understanding of the underlying mechanism, and to develop efficacious, stable and hypoglycemia-risk free drugs, it is crucial to gain more knowledge about diabetes from animal models. In this review, the types of diabetes animal models, modeling methods, the advantages and disadvantages, their applicable scope are discussed aiming to provide a reference for researchers to choose appropriate animal models.

Abstract:Objective To summarize the ADHD animal model and analysis the advantages and disadvantages of various models, providing reference to establish ideal animal model for ADHD and providing new way for the future study of ADHD. Methods Collecting ADHD animal models related literature, analyze the sources of the various models and application scope, summarizes the advantages and disadvantages. Results ADHD model mainly divided into four types: genetic model, the nerve injury model, artificial screening model and environmental adaptability. Four types of model modeling purpose is different, different application scope, advantages and disadvantages with each other. Conclusions According to the relevant evaluation standard, there is no ideal animal model of ADHD. SHR rats is the relatively perfect model. The animal model should be selected according to the purpose of the study screening, establishing the ideal ADHD animal model is the important direction of the current research. direction.

Abstract:Hepatitis B virus (HBV) is an important pathogen threatening to human health. Up to date, various of cell infection models and animal models for HBV and the host are widely used in the exploring research of infection mechanism, new drug development and effective therapeutic method for HBV. However, these models have some defects, such as low infection rate, rather short infective stage, and comparatively large species differences with human, and so on. Among them, the biggest problem is that these models cannot completely simulate HBV infection process and pathological changes naturally occurred in human. Herein, the major HBV infection models developed in the past fifteen years, as well as the latest research progress, are presented as a brief review, to provide a reference for constructing novel HBV infection models in the future.