Abstract:The Collaborative Cross(CC) is a powerful genetic resource with a wide range of applications in medical research. It is a multiparental genetic reference population, composed of 200 recombinant inbred mouse strains. It was designed to be a resource for rapid mapping of genes that mediate complex traits. The CC has been produced following an intense breeding program that had taken over ten years to complete. It is now becoming available to the research community. While advances in human genetic technologies over the last decade have been substantial, the CC provides the ability to make discoveries not possible with other methods or resources. It can be applied to any subject that can be measured or modelled in the mouse. Here, we discuss a range of applications for which the CC can be used, with examples taken from the early characterization of this powerful resource.

Abstract:Collaborative Cross mice (CC mice) are series of inbred mice strains generated from hybrid strains of mice with different genetic background which used for human complex diseases and genetic diversity diseases studies. Genetic diversity of CC mice can reflect different mouse subspecies, the single nucleotide polymorphism is four times than traditional inbred mice. CC mice are more and more widely used in the field of life science and medical research. Based on information retrieval of CC mice, we introduced the related information resources of CC mice origin, database, application tools, and research results, to promote CC mice resources to be used widely in China.

Abstract:Objective To investigate the effect of cyaniding-3-glucoside (Cy3G) on glucose and lipid metabolism in the APP^swe/PS1^ΔE9 mouse model of Alzheimer's disease (AD). Methods Seven-month-old APP^swe/PS1^ΔE9 (PAP) mice were randomly divided into model group (PAP), Cy3G treatment group (PAPCy, 5 mg/kg/d) and negative-control group (nPAP). In addition, age-matched and normal wild-type of C57BL/6J mice were selected and divided into vehicle group (WT), Cy3G intervention group (WTCy, 5 mg/kg/d). Each group containing 12 mice, with equal number of male and female mice. After 8-week Cy3G supplementation, microPET/CT was used to measure cerebral glucose metabolism rate of mice in each group. Biochemical methods were used to detect the liver/kidney function as well as indicators associated with lipid metabolism. After weighting brain tissue, the brain coefficient was tested and pathological examination was used to observe tissues changes. Transmission electron microscope was used to observe neuropathological amyloid plaques deposition. Western-blot was used to determine protein levels of AKT and JNK. Results Compared with the WT group, PAP mice had low levels of ¹⁸F-FDG uptake rates, especially in the regions of the frontal lobe and hippocampus accompanied by the decreased brain coefficient and amyloid plaques deposition in hippocampus. And levels of aspartate transaminase (AST) and lactic dehydrogenase (LDH) were also increased in PAP mice, but lipid metabolism index was relatively normal. In addition, the expression of JNK was decreased and AKT was increased in mice of PAP. However, in the PAPCy group, ¹⁸F-FDG uptake rates were obviously increased in the regions of the frontal lobe and hippocampus compared with those in the PAP mice. And the reduction of brain atrophy and amyloid plaques deposition, normal lipid metabolism and no obvious liver/kidney toxicity were also observed. Cy3G also could reverse the changes of JNK and AKT protein. Conclusions Cy3G can improve glucose metabolism disorders instead of lipid metabolism, inhibit the senile plaques deposition in hippocampus and regulate insulin resistance and inflammatory reaction associated with JNK/AKT pathway. Thus, Cy3G has a good safety profile and may be used as an ideal alternative to traditional disease-modifying treatments against AD.

Abstract:Objective To evaluate the safety of Light emitting diode (LED) source through irradiating the skin of Japanese big-ear white rabbits. Methods Animals were randomly divided into irradiated group and control group. Animals in the two groups were irradiated with LED light source after eliminating back hair (50mw/cm²) and the ordinary fluorescent lamp, respectively. The animals were irradiated for 5 hours every day for 3 months. The changes of animal skin symptoms, feeding, body weight, and temperature were observed and detected; the blood routine and biochemical indexes were tested; the changes of haematological immune cells and cytokines were analyzed by ELISA; histopathology and elastic fibers of skin were detected; the levels of C-myc, P53, and cyclin D1 in skin were tested by immunohistochemistry. Results The skin of rabbits in irradiated group showed no abnormal change; the weight was higher than the control group animals at the 6th week, and the temperature had difference at the 3rd and 6th week, the diet showed no abnormal. Blood routine showed white blood cells increased; biochemical detection manifested urea and creatinine levels increased, but the indexes are in normal range; the hematological CD3T cells, cytokines IL-6,TNF-α and IFN-γ in the two group animals had no significant difference. The levels of CD19B cell and IL-4 level increased in irradiated group; The viscera histopathology structure and skin elastic fibers distribution in irradiated group animals showed normal; the expression of cell cycle protein 1 (CCND1), c-myc protein and P53 protein in skin of irradiated group had no difference with control. Conclusions After irradiating 3 months, the LED light source had not caused the obvious pathology damage of tissues and change of skin tumor related factors to the rabbits.

Abstract:Objective To construct the eukaryotic expression vector GV394-Nurr1 containing human Nurr1 gene and to study the effects of transient transfection of Nurr1 on intracellular reactive oxygen species level. Methods The full-length of human Nurr1 gene amplified by PCR was subcloned into T vector and sequenced. GV394-Nurr1 vector was constructed by BamHI and XhoI double digestion and then T4 DNA ligase conjunction. GV394-Nurr1 was transfected into SH-SY5Y cells by liposome transfection technique; The mRNA of Nurr1 was detected by RT-PCR; The effect of Nurr1 expression on intracellular reactive oxygen species (ROS)was detected by (DCFH-DA) staining.Results PCR and sequencing confirmed that the Nurr1 gene was correctly cloned into eukaryotic expression vector GV394. The RT-PCR results showed that the Nurr1 mRNA expression in the neuroblastoma SH-SY5Y cells transiently transfected Nurr1 was higher than that in the control group. DCFH-DA staining showed that the level of reactive oxygen peak in neuroblastoma cells transiently transfected Nurr1 obviously shifted to the left compared to the control group. Conclusions The human Nurr1 gene eukaryotic expression vector was successfully established and its high expression in the neuroblastoma SH-SY5Y cell line significantly decreased the ROS level. This provide the basis for further study on the function of Nurr1 in vitro and its relationship with the protective effect of dopaminergic neurons.

Abstract:Objective To investigate the bcl-2 gene modification on neurological function recovery in rats with spinal cord injury in neural stem cell transplantation. Methods Cultured rat neural stem cells by Ad-EGFP as vector-mediated side B-cell lymphoma 2 gene (bcl-2) gene transfection of neural stem cells were divided into 3 groups:control group, negative transfection group, bcl-2 transfection group. Use western-blot to detect the expression of bcl-2 protein in neural stem cells before and after transfection. 85 adult female SD rats, successful model 72, were randomly divided into control group, NSCs group, bcl-2-NSCs groups, 24/group, rat acute spinal cord injury model in accordance with a modified Allen's method. Assess the motor function by BBB rating and the swash plate test. 7 days after modeling by RT-PCR and Western blot detection of spinal cord injury around HSP27, c-fos gene expression, TUNEL assay apoptosis. Four weeks after model drawn line HE staining and fluorescence microscopy EGFP-labeled NSC survival and distribution of the rats neurophysiological recovery by SEP and MEP.Results bcl-2 gene transfection of rat neural stem cells, bcl-2 transfection group and control group, negative transfection group compared to bcl-2 mRNA and protein levels were expressed (P<0.05); lower extremity motor function in rats evaluation of bcl-2-NSCs group than NSCs group, NSCs group than the control group. 72 hours after modeling, bcl-2-NSCs number of apoptotic cells were significantly lower than the control group and NSCs group (P<0.05). 7 days after modeling, compared with the control group and NSCs group, bcl-2-NSCs group HSP27 gene and protein expression was significantly higher than that (P<0.05), bcl-2-NSCs group c-fos mRNA and protein expression was significantly reduced compared (P<0.05). 4 weeks after modeling, HE staining control group showed spinal cord tissue loss and the formation of syringomyelia, no axonal through. NSCs group damage zone few of neuraxis-like structures, syringomyelia smaller, bcl-2-NSCs group showed more nerve axon-like structure, no syringomyelia. EGFP-positive cells labeled:bcl-2-NSCs group the most, NSCs group followed, no control group, and the difference between the groups was statistically significant (P<0.05). After the 4th week, SEP and MEP latency period:bcl-2-NSCs group P<0.05); Volatility:bcl-2-NSCs group> NSCs group> control group, and between the groups was significant difference (P<0.05). Conclusions By Ad-EGFP as vector-mediated side B-cell lymphoma 2 gene (bcl-2) gene transfection make neural stem cells can promote cultured rat neural stem cells. bcl-2 gene-modified neural stem cell transplantation can promote the regeneration of spinal cord injury synaptic elevated HSP27 expression after spinal cord injury, reduced expression and neural cell apoptosis after spinal cord injury bcl-2 gene and improve limb movement in rats function and electrophysiological function.

Abstract:Objective To investigate the Protective effect of propofol on the retinal ganglion cells of the rat optic nerve crush model. Methods 67 SD rats. Randomly selected 20 rats, don't do any processing for the normal group. With more than 47 rats optic clamps for optic nerve contusion model, legal system building 5 failure, success of 42 rats were randomly divided into the optic nerve damage and propofol group, 21/group. Will not be any treatment after optic nerve injury group building, 4 hours after propofol group for propofol therapy. After 4 days of successful modeling, the apoptosis of retinal ganglion cells were detected by TUNEL staining. After 7 days of successful modeling, the expression of Caspase-3, BCL-2 in retina and optic nerve cells of rats were detected by RT-PCR and Western-blot.After 14 days of successful modeling, The amplitude and latent period of P1 wave of flash visual evoked potential were detected.The animals were sacrificed and the optic nerve was taken to observe the pathological morphology of retina and optic nerve in rats, and the retinal ganglion cells (RGC) were counted. Results After 4 days of successful modeling,the apoptosis of the propofol group was significantly lower than that of the optic nerve injury group (P<0.05).After 7 days of successful modeling,the expression of Caspase-3 in the propofol group was significantly lower than that in the propofol group (P<0.05), the expression of BCL-2 in the propofol group was significantly higher than that in the propofol group (P<0.05).After 14 days of successful modeling, FG Positive RGC numbers:normal group>propofol group> optic nerve injury group and between groups difference was statistically significant (P<0.05). The flash visual evoked potential of the rats in propofol group was significantly shorter than that in the optic nerve injury group (P<0.05) and the amplitude of the visual evoked Potential was significantly higher than that of the optic nerve injury group (P<0.05).Conclusions Propofol treatment can through an early reduction in rats after optic nerve crush RGCs apoptosis, decreased Caspase-3 expression, increased BCL-2 expression and improve number of optic nerve crush of RGCs survival.

Abstract:Objective To explore dezocine post conditioning on acute lung injury induced by intestinal ischemia and reperfusion induced in rats. Methods According to the random number seed, the healthy male SD rats were randomly divided into 4 groups:Rats in Control group (CON group) were subjected to separate the superior mesenteric artery without being obstructed. Rats in intestinal ischemia-reperfusion group (Ⅱ/R group) were set intestinal ischemia-reperfusion model and administered saline 0.6 mL intravenously after occlusion. In dezocine post conditioning group (Dez group)rats were administered dezocine 0.6 mL 3 mg intravenously after occlusion. In 5-hydroxydecanoate sodium group (5-HD group) rats were administered 5-hydroxydecanoate sodium (10 mg/kg) by peritoneal injection 30min before occlusion then operated as group Dez. The animals were killed at 1 h of reperfusion with the intestinal tissue removed to calculate the index of quantitative assessment of intestinal mucous membrane injury. Tissues of left lung were obtained for observation of histopathology with light microscope and the index of quantitative assessment of histologic lung injury was calculated. The concentrations of malondialdehyde (MDA), superoxide dismutase (SOD) activation and myeloperoxidase level in each group were detected via tissues of left lung. The concentrations of TNF-a and IL-6 were also examined. Results Acute lung injury induced by intestinal ischemia and reperfusion induced in rats was improved by dezocine post conditioning. Chiu score, lung injure score, malondialdehyde (MDA) content, myeloperoxidase(MPO) level and the concentrations of TNF-a and IL-6 were descended while superoxide dismutase (SOD) activation was rised. The protective effects of dezocine post conditioning were attenuated by injecting 5-hydroxydecanoate sodium via peritoneal cavity which was a selective mitochondrial ATP-sensitive potassium channel antagonist. Conclusions Dezocine post-conditioning protects against lung injury induced by intestinal ischemia-reperfusion in rats. One of the protective effects may be related to opening mitochondrial ATP-sensitive potassium channel.

Abstract:Objective To investigate the role of small RNA 195 (MicroRNA-137), TGF-β1/Smads signal transduction pathway and angiotensin Ⅱ (Ang Ⅱ) in cardiac remodeling in spontaneously hypertensive rats (SHR). Methods 16 SHR male rats were randomly divided into intervention group SHR (captopril 10 mg/kg·d) and SHR control group (distilled water), the other 8 Wistar rats were normal control group, rats were given captopril 10 mg/kg·d or distilled water for 8 weeks. Caudal arterial pressure was measured before and after the intervention, intervention after 8 weeks rats were killed by exsanguination, HE staining was used to observe the morphological changes of rat heart, qRT-PCR method was used to detect the expression of miRNA-137 in rat heart, Western-blot detection of TGF-β1 and Ang Ⅱ, Smad 3, Col-Ⅰand Col-Ⅲ protein. Results Compared to the normal control groups,the miRNA-137,AngⅡ,TGF-β1,Smad3,Col-Ⅰand Col-Ⅲ were higher expressed in SHR treatment group and SHR control groups(P<0.01 or P<0.05); Compared to the SHR control group,the cardiomyocyte of SB group becomes smaller and arranged more closely and orderly,the miRNA-137,AngⅡ,TGF-β1,Smad3,Col-Ⅰand Col-Ⅲ were significantly lower expressed(P<0.01 or P<0.05). Conclusions MiRNA-137 may promote SHR cardiac remodeling by up regulation of Ang Ⅱ and TGF-β1/Smads signaling pathway; the captopril intervention can inhibit miRNA-137 expression.

Abstract:Objective To explore effect of miR-126 on apoptosis of myocardial cells in rats with acute myocardial infarction (AMI). Methods AMI model was established by ligation of left anterior descending branch of coronary artery. The survivors were randomly divided into 3 groups:AMI group, NC group and miR-126 group. the NC group and miR-126 group were injection of lentiviral transfection of miR-126 mimics NC and miR-126 mimics. The sham operation group was left anterior descending coronary artery without ligation. The rats' cardiac function was recorded. Apoptosis index and infarct size of myocardium was detected by TCC method and in situ end method (TUNEL), respectively. The activity of cysteinyl aspartate specific proteinase 3 (Caspase 3) and Caspase 8 were determinated by colorimetry. The expression of Bcl-2, Bax was assayed by Western-blot. The expression of Fas and Fas-L mRNA was detected by RT-PCR. Results Compared with sham operation group, left ventricular ejection fraction (LVEF) and left ventricular long axis shortening fraction (FS) was increased, left ventricular end diastolic diameter (LVDd) and left ventricular end systolic diameter (LVDs) was decreased, apoptosis index was increased, myocardial infarction area increased, the activity of Caspase 3 and Caspase 8 was increased, the expression of Bax protein was upregulated, the expression of Bcl-2 protein was downregulated, the expression of Fas and Fas-L mRNA was increased in AMI and NC group, the difference was statistically significant (P<0.01). Compared with AMI and NC group, miR-126 could reverse the change, the difference was statistically significant (P<0.01). Conclusions miR-126 could inhibit apoptosis of myocardial cells in rats with AMI, which related to regulation of expression of cell apoptotic related protein.

Abstract:To design and make a simple fixture device for holding mice in rear paws experiments as a safe and convenient experimental tool for sweating and hidroschesis experiments.

Abstract:Locomotion is one of the most vital and fundamental motor behaviors in rats, which can reflect the performance and characteristics of motion in various experimental animal models, it has an important clinical significance to motor dysfunction rating and rehabilitation effect evaluation grade for developing an evaluation method and standard of locomotion in rats. This article will comprehensively analysis and review the literatures of locomotion in rats on the evaluation methodology and typical application in order to provide necessary reference for researchers.

Abstract:P2X3 subunit was one of the P2X receptor family members, which highly selective expressd in the nociceptors, and involved in the pathological process of inflammatory pain. Currently, animal inflammatory pain models included formalin inflammatory pain model, Freund's adjuvant inflammatory pain model, the carrageenan inflammatory pain model, bee venom inflammatory pain model,et al.Different inflammatory pain model had different pathological features, the main features of the formalin pain model is double phase of spontaneous pain, due to different duration time of inflammation, carrageenan inflammatory pain model is commonly used in sub acute inflammation model, Freund's adjuvant inflammatory pain model, bee venom inflammatory pain model, and turpentine inflammatory pain model are for chronic inflammatory pain model research, the expression and function of P2X3 may be different.This paper focused on the effects of P2X3 receptors in different inflammatory pain models.

Abstract:miRNAs are a class of small endogenous RNAs that degrade target mRNAs or repress their translation process. Several miRNAs in glioma are up-regulated, while some others down-regulated. Some miRNAs promote tumorigenesis; some others, however, play a similar function of tumor suppressor genes. Therefore, studies on the expression profiles of miRNAs in glioma may afford auxiliary basis for early clinical diagnosis and novel srtategies for therapy of glioma. This paper will review on researches about the expression levels of miRNAs and their targets in glioma.

Abstract:Behavior assessment of cognitive function has been widely used in related research on disease models, especially in physiological mechanism and drug intervention evaluation. Evaluating the function of learning and memory is the most common one in colourful behavioral experiments.The thesis summarizes the methods of analyzing the common cognitive behavior about animal models in recent years, briefly introduces the comment contents, advantages and limitations of each behavior test, and also provides a reference for behavior analysis mainly to the related diseases model of cognitive dysfunction.

Abstract:The combination of medical laboratory animal science teaching with standardized certificate training is an effective reform for the laboratory animal science teaching of postgraduates. On the one hand, by the changing of laboratory animal science teaching mode, which can combine the general knowledge of laboratory animal science with their respective specialty, it is able to improve their practical ability to carry out animal experiments, the ability to design and implement independent research and to innovate. On the other hand, by the learning of the national law, regulations and relevant ethical knowledge, the students can acquire the certificate, which is the threshold for processing the laboratory animals, and ultimately achieve precise and reliable experimental results, normalized and standardized method, proceeding in line with the ethical principles.