Abstract:Objective We established the animal models of obesity induced by high-fat diet, in order to study the mRNA and protein expression of regulation molecules related with iron metabolism about hepcidin, lipocalin-2 (LCN2), ferroportin-1 (FPN1) in obese mice's liver and the molecular regulation mechanism. Methods C57BL/6J (4～6 weeks) mice were randomly divided into control group and obesity model group, each group of ten. The obesity group were fed with a high-fat diet and the control group were given the normal diet for lasting 15 weeks. After we successfully established the obesity animal model, the expression level of hepcidin, LCN2 and FPN1 mRNA in the liver were measured by Real-time fluorescent quantitative PCR method and the protein expression level of LCN2 and FPN1 were measured by Western-Blot. Results Compared with the control group, the expression level of hepcidin mRNA in the liver was increased in obesity group (P < 0.05), however, the expression level of LCN2, FPN1 was no significant difference (P > 0.05). Conclusion Obesity can increase the expression of hepcidin mRNA, however, there was no significantly effect on the expression of LCN2, FPN1. So, we can't think that obesity can affect the expression of LCN2 and FPN1, lead to the ability of cells uptake and release iron abnormal, then appear iron metabolism disorders. As a result, leading to iron deficiency. Maybe obesity can affect other regulatory molecules related with iron metabolism through up-regulation the expression of Hepcidin or the more complex regulatory mechanisms. We still need further experimental research and exploration. This research also provides the basis of theoretical and experimental for the further study the effects of obesity on the expression of regulation molecules related with iron metabolism in obesity mice's liver and the mechanism of iron deficiency.

Abstract:Objective Investigate the effect of insomnia on cardiovascular disease factors and offer the experimental evidence for treating cardiovascular disease with traditional Chinese medicine tranquillization methods. Methods Sixteen Wistar rats were randomly divided into two groups named sleep deprivation (SD) group and normal control check(CC) group. Body weight and electrocardiogram were recorded and serum concentrations of melatonin (MT), endothelin-1 (ET-1), IL-6 and TNF-α were tested before SD and 2 days, 5 days, 7 days after SD. Results Body weight decreased in SD group while increased in CC group. Compared with CC group, body weight of SD rats decreased significantly in 5 days and 7 days after SD(P < 0.05). Compared with CC group, TNF-α increased significantly in 5 days (P < 0.05). With the time, heart rate accelerate and QTc were prolonged, MT decreased while ET-1, IL6, TNF-α increased significantly in 7 days after SD (P < 0.05). Conclusion Long term insomnia would decrease body weight and MT, while increase heart rate, QTc, ET-1 and inflammatory factors, which increase cardiovascular disease factors. It provided the experimental evidence for the study on traditional Chinese medicine tranquillization methods in the treatment of cardiovascular disease.

Abstract:Objective To develop a model that could copy the pathological development of psoriasis, the triple-transgenic mice that harboring Plasminogen activator, urokinase (PLAU),PLAU receptor (PLAUR) and signal transducer and activator of transcription 3 (STAT3) were generated. They are the important genes involved in the pathological development of psoriasis. Methods The transgenic plasmid was constructed by insertion of the PLAU, PLAUR and STAT3 into the downstream bovine keratin 5 promoter respectively. The transgenic mouse was produced by microinjection and the genotyping was detected by PCR. The expression level of the transgenic gene was determined by Western blotting. The pathological changes were observed by HE staining. Results One mouse line was selected with over expression of the PLAU, PLAUR and STAT3 in the tissue of skin. The transgenic mice showed decreased dermal layer, a hyperkeratinized cuticular layer and increased stratum spinosum. The number of hair follicle was reduced and developed abnormally in the transgenic mice. The Munro abscess in the dermal layer and the increased inflammatory cell infiltrates in dermal layer were also observed in the transgenic mice. Conclusions A transgenic mouse line was produced and passage stably, which expressed the PLAU, PLAUR and STAT3 in the tissue of skin and developed the psoriasis progressively. All of our results suggested that the transgenic mice were a useful animal model for psoriasis.

Abstract:Objective To investigate the changing characteristics and rules of the different period of articular cartilage in rats after dorsal rhizotomy, and to verify the partial sensory disturbance can cause articular cartilage injury. Methods Thirty-three ten-month-old SPF male Wistar rats were randomly divided into experimental group (n=18) and control group (n=15). The rats in the experimental group were sectioned the L3、L4 dorsal roots in the right. The rats in the control group were only incised the skin and paravertebral muscles. To observe the behavior changes in rats. At 2, 6 and 10 weeks, the specimens of the right hind lower end of femur were drawn out to make paraffin sections, then HE staining and Safranin O/ Fast Green staining. The changes and characteristics of the morphology of the articular cartilage was observed. Results With the prolonging of period, the right hind limb of the experimental rats through the change process of transient paralysis, coordination of movement disorders and active movement. In the experimental group, the patellar surface of right hind femur gradually became shallow and wide. The articular cartilage underwent rough, cells disorganizated, cells decreased, and duplicated drifted and interrupted tide line. The ratios of ACC/TAC of the experimental group were gradually high(t=5.25～8.13,P < 0.05). Conclusion Dorsal rhizotomy can cause injury and degenerative changes in articular cartilage.

Abstract:Objective To explore the impacts of cSNP on the structure and function of rhesus macaque's BST-2. Methods Extracting RNA from peripheral blood of rhesus macaques, then RT-PCR, Monoclonal sequencing to find the cSNP sites; Forecasting the structure of these proteins; Comparing the Level of SIVmac239 replication between the different genotypic Rhesus macaques. Results We found 8 non-synonymous mutation sites, in the 8 non-synonymous mutation sites, Only G41A, T128C, C129 and A333C change the secondary protein structure of BST-2 by forecasting of Psipred software; At the plateau of SHIVSF162p3 replication, The SHIVSF162p3 replication level of reference genotype rhesus macaque (DLQ) is higher than rhesus macaques of GLQ genotype, other genotypes rhesus macaques have no significant difference. Conclusion cSNP (G41A) may influence the antiviral activity of rhesus macaque's BST-2, this study gives us a reference to further research work.

Abstract:Objective To study the impact of aging on the capability of lung stem cell steady-state maintaining and bronchial epithelial cells regeneration and differentiation during the repair of lung epithelial cells after naphthalene induced bronchial epithelialium injury. Methods The proportion of lung stem cells in mice after naphthalene treatment was analyzed by immunohistochemistry and FACS. The repair efficiency of lung epithelial cell in young and old mice was examined by immunohistochemistry staining and FACS. Results The data suggested that aging didn't change the proportion of lung stem cells (including the distal lung epithelial stem cells/progenitor cells and lung mesenchymal stem cells/progenitor cells) under normal physiological conditions. After naphthalene injury, more serious injury and decreased repairing capacity was observed in old group. Lung progenitor cells /total lung cells decreased during the repair process of lung bronchial epithelialium (clara cell) injury. The ratio of regenerated cell to lung progenitor and stem cells were also significantly decreased in old group. Conclusion The regenerated capability of lung stem cells after lung bronchial epithelialium injury decreased with aging. This might be the reason of more incidence of lung injury and worse therapeutic results in the elder in clinic.

Abstract:Objective To make comparisons of the three models of acute and chronic rheumatic carditis to find out an optimal animal model. Methods AntigenⅠwas a emulsifier mixed by complete freund's adjuvant(CFA) and Group A streptococcus(GAS). AntigenⅡ was mixed by incomplete freund's adjuvant(IFA) and GAS. Female Lewis rats were randomly divided into four groups: A, B, C treatmeat groups were immuned with antigenⅠat the foot pad firstly. Subsequently, rats in group A、B、C were injected antigenⅠ, antigenⅡand activated GAS respectively to make the models of RHD. Rats in control group D were immunized with the same protocol outlined as treatment groups but without GAS. Respectively 7, 12, 24 weeks the rats were sacrificed 24(each group was 6). The blood biochemical item and Hematoxylin-eosin(HE) staining of hearts were detected. Results In group C the mortality was 25%. In group A, the incidence of carditis was the highest. Histopathological manifestations of group A, C was not only revealed acute damage such as inflammatory cell infiltrate as well as group B, but also the Aschofflike cells in the myocardial cells interstitial. But in group A and C there had a great degree of the inflammatory cells infiltration than group B. At 24th week rats in group A detected the rate and degree of valve fibrosis in chronic damage were higher than group B and C. None of rats in group D presented carditis or valvulitis. Conclusion In group A, giving the GAS with continuous stimulation after using the mixed emulsification of CFA and GAS to immune Lewis rats for five times was a appropriate method which could provide an optimal animal model for experimental study of acute and chronic rheumatic heart disease.

Abstract:Objective Analysis of the genetic structure stability of Brandt's vole (lasiopodomys brandtii) in closed colony using microsatellite marker technology. Methods Genomic DNA was extracted from tail tip using high-concentration-salt precipitation methods. Marked with fluorescent tags(Fam), 7 microsatellite primers were filtered out by PCR, and the DNA structure of three consecutive generations of Brandt's vole was analyzed by microsatellite marker. Results By the analysis of the average heterozygosity and polymorphism information content, Brandt's vole populations maintaineda closed group of qualified genetic structure. Conclusions The present results show that the closed group of Brandt's vole species in our laboratory maintain a stable genetic structure.

Abstract:Objective To observe the Interference effects of siRNA(small interference RNA) intrathecal injection on the expression of mRNA and protein of MrgC, on PWTs (paw withdrawal thresholds) and the phosphorylation of PKCε Ser729 in ipsilateral DRG(dorsal root ganglion) of rats with chronic inflammatory pain induced by CFA(complete freund's adjuvant). Methods 16 health adult male SD (Sprague-Dawley) rats were randomly divided into 2 groups: control siRNA group and MrgC siRNA group, 8 rats in each group. After success of intrathecal catheterization, corresponding siRNA was injected in rats for 4d, once a day, 5μg/d per rat. The model of chronic inflammatory pain was established by CFA (0.1ml per rat) subcutaneously injected into the right hind paw at 4th day post-administration, then two groups were administrated corresponding siRNA on alternate day and executed at the 11th day post-administration. The PWTs were measured at 5 time points of pre-intrathecal catheterization, pre-administration, 4th day post-administration(0h post-CFA injection), 5th day post-administration(24h post-CFA injection), 11th day post-administration(7 d post-CFA injection). The expression of MrgC mRNA in ipsilateral DRG was detected by fluorogenic quantitative PCR, and the expression of MrgC and the phosphorylation of PKCε Ser729 in ipsilateral DRG was detected by immumofluorescence method. Result Compared with 4th day post-administration, PWTs of both two groups at 5th day post-administration decreased significantly (P<0.01). While there was no significant difference of PWTs between two groups at every detective time point. Compared with control siRNA group, the expression of MrgC mRNA and the rate of MrgC positive cells in MrgC siRNA group both decreased significantly (P<0.01,P<0.05), whereas the rate of p-PKCε Ser729 positive cells increased obviously (P<0.05) at 11th day post-administration. Conclusion MrgC siRNA can effectively interfere the expression of mRNA and protein of MrgC in L4-L6 ipsilateral DRGs of rats with chronic inflammatory pain induced by CFA, and the siRNA interference on MrgC can obviously up-regulate the phosphorylation of PKCε Ser729, while it has no significant effect on PWTs of rats.

Abstract:Objective To evaluate the effects of baijin capsule on behavioral changes and monoamine neurotransmitters concentration in chronic unpredictable mild stress (CUMS)depression rat model. Methods The depression rat model was induced by11-week chronic unpredictable mild stress combining with solitary. After the model were established, rats were given the decoction of baijin capsule (12.6 g/kg, 4.2 g/kg, 1.4 g/kg) or fluoxetine hydrochloride (3.5 mg/kg) by intragastricfor 4 weeks. During the experiment period, sucrose consumption and open-field experiment were conducted to monitor the behavior of rats, such as sucrose consumption percentage, horizontal motion, and vertical motion. At the end of the experiment, the levels of the monoamine neurotransmitters in the cerebral cortex and hippocampus were analyzed by method of high performance liquid chromatography-electrochemistry. Results Compared with the normal group, the weight, horizontal displacement distance, vertical movement times, and sucrose consumption percentage of rats in model group decreased significantly after stimulated with CUMS and solitary for 7 weeks (P<0.05,P<0.01).Compared with model group, consecutively administrated for 4 weeks, horizontal displacement distance, vertical movement times, and the percentage in sugar water consumption significantly increased with the treatment of baijin capsule (P<0.05, P<0.01). Meanwhile, the content of norepinephrine (NE), dopamine (DA), and 5-hydroxytryptamine (5-HT) in the cortex were significantly increased in rats of the baijin capsule (P<0.05). Conclusions The results indicated that baijin capsule improved the behavioral disturbances in depression rat model, which were related to enhancement of the concentration of monoamine neurotransmitters in the cortex.

Abstract:Objective The goal of this study is to understand the function of FKBP51 in resistant to high fat diet-induced obesity using FKBP51 knockout (KO) mice and in vitro adipocyte differentiation. Methods Four-week old male FKBP51 KO and wild type (WT) mice were fed separately with regular or high fat diet for 6 weeks. The body weight and food consumption were recorded weekly, the energy expenditure differences (O₂ consumption, CO₂ production, respiratory exchange ratio, and heat production) of each group were monitored using the MM-100 metabolism cages system for 24 hours, then the liver from the above animals were stained with the Oil red-O to detect the lipid accumulation and the expression of metabolic genes. In addition, induction of adipocyte differentiation of immortalized MEF cells from WT and FKBP51 KO mice were used to observe the effect of FKBP51 gene on lipogenesis. Results Compared to WT mice, FKBP51 KO mice has less weight increment, and less lipid accumulation in the liver, but with no difference on food consumption during high-fat diet fed. Moreover, FKBP51 KO mice exhibited more O₂ consumption, CO₂ production and heated production under both RD and HF diet conditions. The PEPCK,G6Pase and UCP-1 genes up-regulation. In addition, lipid content was reduced in FKBP51 gene deficient MEF cells after adipocyte differentiation. Conclusions The FKBP51 gene plays an important role in high fat diet-induced obesity through the energy metabolism enhancement and lipogenesis inhibition.

Abstract:Objective To optimize the scheme of Mongolian gerbil superovulation. Methods Based on the analysis of the animal-week-old, the dose and time interval of hormone, we got the best animal-week-old, dose and time interval of hormone for Mongolian gerbil superovulation. Results The 6 week old female Mongolian gerbils which injected of 10 IU PMSG and followed by 10 IU hCG in 70 hours later could get the best superovulation. We collected eggs at 16 hour after mate with male gerbils. The ovum pick-up rate reached 80%, the number of oocytes were 32.6±3.0, the number of the fertilized egg developed to 2-cell are 24.8±5.4. Conclusions This study summarizes the optimization scheme of Mongolian gerbil superovulation induced by PMSG and hCG and it supported the foundation for Mongolian gerbils embryo biotechnology.

Abstract:Objective To explore the best sampling position of bone marrow biopsy in mice. Methods Mice were sacrificed by cervical dislocation, then take the skull, sternum, tibial, femoral and iliac bone, making pathological section. Observed and photographed under light microscope. Comparison of the distribution of the essence and the hematopoietic microenvironment in different parts. Select site which can best reflect the hematopoietic function of bone marrow. Results A large number of hematopoietic cells in ilium marrow sections visible. The cells are evenly distributed. Blood and megakaryocytes were clearly visible. The arrangement of the structure of the scaffolds for tissue closely. The number of fat cells less. Bone marrow hematopoietic cells in the skull, sternum, tibial, femoral were not clear and not active. And there are more fat cells. Conclusions As the best sampling position of bone marrow biopsy, is ilium.

Abstract:Experimental pathology is an important part of life science research associated with animal experiment. Acquisition and fixation of optimum specimen and subsequent section of paraffin embedded tissue and dyeing are key factors playing important role in reliability, authenticity of pathological diagnosis. This paper summarizes the problems encountered in pathological section making of animal experiment and it correspond solutions.

Abstract:Adipose-derived stem cells(ASCs) as potential seeded cells have been widely used in tissue engineering. Thus to obtain enough, high activity, high purity adipose-derived stem cells is the particular important premise of the application in tissue engineering. In this paper, the isolation and purification methods of ASCs were reviewed and the merit and demerit of different methods were compared in order to provide theoretical basis for safe and high-effective isolation and purification of ASCs.

Abstract:PD-1:PD-L1/PD-L2 pathway played negative immune role in most tumor and several infectious diseases, but the immune role in tuberculosis (TB) was not explicited. Since the high expression of PD-1:PD-L1/PD-L2 pathway was correlated with MTB infection both in vitro and in vivo, the in vitro negative immune role was indicated by inhibited T cell protective immune function, while the in vivo immune role was not clear. The binding of PD-1 and PD-L1 played an immunosuppression role, the expression of PD-L1 was regulated by Th1-type cytokines. The binding of PD-1 and PD-L2 played protective immune role, and the expression was regulated by Th1-type cytokines. The immune role and mechanism of PD-1, PD-L1 and PD-L2 would be investigated by Cas9-CRISPR technique, which would provide theoretical basis for the tuberculosis prevention and control.

Abstract:The pathogenesis of multiple sclerosis (MS) involves alterations to multiple pathways and processes, which represent a significant challenge for developing more-effective therapies. In MS, abnormalities have been identified in several cytokine-signaling pathways, as well as those of other immune receptors. Among the downstream molecules implicated are Jak/Stat, NF-κb, ERK1/2, p38 or Jun/Fos, current MS drugs target some of these pathways. This article will with the aid of the latest research results of systems biology approaches that study pathway dysregulation in the process of MS development, targeting these relevant MS-signaling pathways, offers the opportunity to accelerate the development of novel individual or combination therapies for the future of new drug research.