Abstract:Objective To explore the feasibility of combining chitosan conduit filled with simvastatin/pluronic F-127 thermosensitive hydrogel to repair peripheral nerve defects in rats. Methods The chitosan conduits and simvastatin/pluronic F-127 hydrogel were prepared and tested by stereomicroscope, SEM, vitro degradation testing and rheometer property. A total of 40 Sprague-Dawley rats were randomly divided into 4 groups (n=10 per group): the chitosan conduit alone (conduit group) or the conduit filled with pluronic F-127 hydrogel containing 0 (c+sim 0mg), 0.5 (c+sim 0.5mg), and 1 mg (c+sim 1mg) simvastatin. A 10mm nerve defect was created in the rat left sciatic nerve and bridged with the chitosan conduit filled with pluronic F-127 hydrogel containing different doses of simvastatin. At 10 weeks after surgery, the regenerated nerves were performed the H&E and TEM to observe the morphological change. The diameters of the myelinated axons, the thickness of the myelin sheath, and the G-ratios (the ratio of the axon diameter to the total fiber diameter) were calculated to make statistical analyses. And the immunohistochemical method was used to measure the expression of NF200 and S100 protein in the regenerated nerves and to detect the expression of neurotrophins including pleiotrophin (PTN), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF) and glial cell line-derived neurotrophic factor (GDNF). Results The chitosan conduits filled with simvastatin/pluronic F-127 promote nerve regeneration in rats. All groups showed the regenerated nerves, but H&E staining showed that the regenerated nerves in the simvastatin treated groups were much thicker than those without simvastatin; And the same trend was also found for the diameter of the myelinated axons, the thickness of the myelin sheath and G-ratio; In addition, the numbers of NF-positive cells indicating regenerated axons and S100-positive cells indicating Schwann cells were significantly larger in simvastatin treated groups than those without simvastatin. And immunohistochemical staining indicated that simvastatin induced an increased expression of PTN, HGF, VEGF and GDNF compared to those found in the control group. Conclusions Taken together, the simvastatin/pluronic F-127 hydrogel filling in the chitosan conduits promoted the reconstruction of histological in peripheral nerve defects, which can be used for peripheral nerve regeneration.

Abstract:Objective To get the full-length HGF,Hepatocyte growth factor, cDNA encoding sequence and predict its molecular characteristics. Methods Full-length HGF cDNA encoding sequence is cloned from total RNA in liver tissue of tree shrew by RT-PCR and its molecular characteristics is analyzed compared with other mammals by using biology software such as DNAMAN,Pymol and so forth. Results A 2193bp of the full-length HGF cDNA encoding sequence is cloned from total RNA in liver tissue of tree shrew.The analysis of molecular characteristics show that the molecular structure of HGF is relatively conservative in evolution, tree shrew HGF overall structure similar to Homo sapiens.However,compared with the structure and charge surface of NK1 domain of HGFα-chain of human,tree shrew has two longer β-sheet and one more α-helix,Additional,tree shrew had a strong positive charged site in this area,which may have a impact on the bind of HGF to its receptor. Conclusion This study provide a theoretical basis for preparation and receptor-binding mechanism studies of tree shrew HGF.

Abstract:Objective To investigate the effect of Slit2 over-expression on nonalcoholic fatty liver disease (NAFLD) in mice. Methods Eight week male LDLR^-/- mouse and LDLR^-/-;Slit2-Tg mouse were fed a high-fat diet for 12 weeks to induce NAFLD(n=6 per group). After 12 weeks, they were taken for blood samples and dissected. The concentration of TG (triglycerides), TC(total cholesterol), LDL-C(low density lipoprotein cholesterol) and HDL-C(high density lipoprotein cholesterol)in the serum were examined respectively. Liver tissue samples were taken for HE staining to observe the morphological and pathological changes. The mRNA expression levels of hepatic genes related to lipid metabolism such as LXR-α,LXR-β FAS,SREBP-1c was measured by real-time RT-PCR. Results The serum LDL-C levels of LDLR^-/-;Slit2-Tg mouse were significantly lower than that in LDLR^-/- mouse(P < 0.01).And the grade of lipid droplets and pathological injury of liver tissue decreased significantly in the LDLR^-/-;Slit2-Tg mouse compared with LDLR^-/- mouse. Furthermore, The mRNA expression levels of hepatic genes related to lipid metabolism decreased significantly in the LDLR^-/-;Slit2-Tg mouse. Conclusions The overexpression of Slit2 can reduce the pathological injury of liver tissue in mouse with NAFLD.

Abstract:Objective To investigate the effects of camel milk on body weight, blood glucose, lipid, insulin and PPAR-γ、TNF-α mRNA expressions in type 2 diabetes rats. Methods Type 2 diabetes was induced by high fat diet and small dose of STZ(i.g. 30 mg/kg), rats were divided into four groups, normal control(control) group, diabetes model(model) group, camel-low dose(CM-L) group(3.5 mg/kg·d), camel-high dose(CM-H) group(10 mg/kg·d). Body weight and fasting blood glucose were measured every week. Glucose tolerance test(OGTT) was conducted at 4th week. After 4 weeks administration, animals were decapitated, plasma lipid(TC、TG、HDL-C、LDL-C), insulin were assayed. The expression of PPAR-γ and TNF-α mRNA were measured using RT-PCR method. Results Compared with control group, the weight of model group lowered significantly(P < 0.01), FBG and blood glucose at 0、30、60、120 min of OGTT, as well as the TC, TG, LDL-C concentrations increased(P < 0.01), while HDL-C content reduced, and insulin level was higher than control rats. Compared with model group, body weight increased and blood glucose decreased in CM group, CM-H reduced the fasting glucose levels in 4th week and decreased the 30min glucose level significantly(P < 0.05). CM lowered the TC, TG, LDL-C, insulin and increased HDL-C concentration, and it is significantly in CM-H(P<0.05). There is a marked decrease in PPARγ mRNA expression(P < 0.05) and a significant increase in TNF-α mRNA expression(P < 0.01)in diabetes rats. Compared with model rats, PPARγ mRNA expression was ascended by CM administration, and it is notably in CM-H group(P < 0.05), while the TNF-α mRNA expression of diabetes rats was lower, but there is no significant. Conclusions CM can improve the lowered body weight, hyperglycosemia, abnormal glucose tolerance and dyslipidemia in type 2 diabetes. The mechanism of these maybe associated with the regulation of PPARγ and TNF-α mRNA.

Abstract:Objective To investigate the protective effect of myricitrin on myocardial ischemia/reperfusion injury in isolated rat hearts and its underlying mechanism. Methods Sixty SD rats were randomly divided into: (1) control group; (2) model group; (3) positive administration group (verapamil of 100 μg/L); (4) three dose groups with varying amount of myricitrin (2.5, 5, 10 mg/L). Each group included ten rats. The myocardial ischemia/reperfusion injury model was constructed using the Langendorff method. The isolated working hearts were ischemia for 30 min followed by reperfusion for 45 min after in equilibrium for 15 min. During the process, we determined myocardial hemodynamic parameters, myocardial enzyme indicators (lactate dehydrogenase (LDH), creatine kinase (CK) and aspartate aminotransferase (AST)) and antioxidative parameters (superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA)). Furthermore, we performed histopathological examination of left ventricles and detected the expression of apoptosis related proteins (e.g., Bcl-2, Bax, Caspase-3 and Caspase-9) by western blot. Results Compared with the model group, myricitrin could significantly improve cardiac constriction and relaxation function, decrease the levels of LDH, CK and AST in perfusate, enhance the activities of SOD and CAT, and decrease the content of MDA in myocardial tissue. The cardiac protective effect of myricitrin was further confirmed by histopathological examination. Apparently, myricitrin pretreatment restrained myocardial apoptosis as evidenced by increasing the level of Bcl-2 expression, decreasing the levels of Bax, Caspase-3 and Caspase-9 expression, and inhibiting the phosphorylation of ERK. Conclusions Myricitrin had a protective effect on myocardial ischemia/reperfusion injury. It showed ability of improving myocardial contractile function, increasing anti-oxidation ability and inhibiting apoptosis. The possible mechanism of cardiac protection of myricitrin was that the agent attenuated myocardial cell apoptosis induced by ischemia reperfusion via inhibiting ERK signaling pathway.

Abstract:Objective The purpose of this study is to understand the function of Fkbp51 in metabolic pathways and neural pathways by profiling gene expression of liver and hippocampus tissues of both Fkbp51KO and WT mice.Methods mRNAs of liver and hippocampus of Fkbp51KO and WT mice were isolated and expression profiling was performed using RNA-seq. Differentially expressed gene between KO and WT were analyzed using BRB-Array Tools. DAVID, STRING, Genecard programs, the Gene Ontology, the Protein-protein Interaction Network and the Gene Annotation were applied to identify significant functional-relevant pathways. Results When compared differentially expressed genes between Fkbp51KO and WT in liver, we found that the loss of Fkbp51 in liver has large effect on the genes related to steroid biosynthetic and metabolic process, lipid biosynthetic process and oxidation reduction. When differentially expressed gene in hippocampus was studied between genotypes, we found that elimination of Fkbp51 has much effect on genes related to detection of mechanical stimulus, learning or memory, regulation of synaptic transmission and the pathway of PPAR and amyotrophic lateral sclerosis (ALS) e.g. When intersection of gene lists between liver and hippocampus tissues, we identified 11 common differentially expression genes. In these genes, HMGCS2 and INSIG1 are grouped in one protein-protein interaction network, and USP2, PER, CRY and DBP are grouped in another network. Conclusions The role of Fkbp51 in metabolism and nervous system is not only independent but also interactive.

Abstract:Objective To explore the possibility of two kind of minipigs as the animal model for metabolic syndrome. Methods Bama minipigs and Tibet minipigs were selected to measure their body weight, body size and metabolic syndrome-related parameters including blood lipid, blood sugar and free radical. Results Compared with Tibet minipigs, Bama minipigs was heavier in weight, shorter in length, larger in neck circumference and abdominal circumference, higher in obesity index, and what's more, the difference in female Bama minipigs was more significant. The levels of TC, LDL-C, HDL-C and TC were higher in female than in male. GLU levels in Bama minipigs was higher than that of Tibet minipigs, and with differences in female Tibet minipigs significantly, while the level of FRA and HbA1c in the Tibet minipig was higher than that of Bama minipigs, Insulin level in female Bama minipigs and male Tibet minipigs was respectively higher than the male Bama minipigs and female Tibet minipigs. MDA levels in female Bama minipigs was significantly higher than that of male Bama minipigs and Tibet minipigs, the level of SOD and GSH-Px in Bama minipigs was higher than that of Tibet minipigs. Conclusions Through the analysis of normal physiological parameters, such as body weight, body measurement, blood lipid, blood sugar and free radical, Bama minipigs tend to be obese; the level of blood fat of female is higher than that of male in two kind of minipigs; the level of blood sugar of Bama minipigs is higher than that of Tibet minipigs, but the ability of Bama minipigs to control blood sugar is better than Tibet minipigs; the ability of Bama minipigs to antioxide is better than Tibet minipigs. In short, according to the specific needs different kinds of minipigs are selected to use in the metabolic syndrome-related research and verified in actual modeling practices.

Abstract:Objective To establish the orthotopic liver transplantation model in gebils. Methods Orthotopic liver transplantation was performed in 200 gerbils with modified two-cuff techniques. The suprahepatic vena cave was anastomosed end to end using nylon suture threat, the portal vein and the infrahepatic vena were inosculated by cuff method, respectively. Biliary tract reconstruction was completed in bile duct stent law. Result After paractise, donor surgery time was 34.56±5.12 min, liver repaired time was 15.43±2.75 min, surgery receptor time was 58.37±8.54 min, no liver time was the success rate (survived more than 6 hours after operation)was 23.66±4.47 min. Conclusion Orthotopic liver transplantation with modified two-cuff techniques in gerbils is a certain feasible. Improving operative techniques is of great importance in increasing successful rate and prolonging the survival time of gerbils after surgery.

Abstract:Objective To investigate the effects and mechanism of acupuncture combined with honeycomb on allergic rhinitis(AR)rat model. Methods AR rat model was induced by ovalbumin(OVA). AR rats were treated with acupuncture combined with honeycomb for ten days. Fluticasone propionate nasal spray was selected as positive control in the text. Behavior analysis were observed and the content of histamine, IgE, IFN-γand IL-4 in serum, the weight of nosal draingage and the number of eosinophils and mast cells, the index of spleen and thymus were detected to evaluate the effects of acupuncture combined with honeycomb on AR rats. Results Compared with model group, acupuncture combined with honeycomb treatment groups displayed markedly lower behavior scores, histamine, IgE and IFN-γ in serum, the weight of nosal draingage and the number of eosinophils and mast cells, the index of spleen as well as higher IL-4 in serum and index of thymus. Conclusion Acupuncture combined with honeycomb displayed significant effects on AR rat model.

Abstract:Objective To investigate the biochemical indicators and pathological changes of early microvascular complications in spontaneous type 2 diabetes (T2DM) ZDF rats. Methods Six male ZDF rats aged around 7-8weeks were allowed to acclimate for 3days after feeding Purina#5008, eight male ZL rats at the same age were assigned to control group, fed with regular diets. At 12,14,16week, blood glucose (GLU), serum cholesterol (CHOL), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), high-sensitivity C-reactive protein (hsCRP), glycated hemoglobin (HbA1c) were detected. All rats were sacrificed at 16 week and pathological examinations were applied to eye, kidney, heart, brain tissues. Results GLU, CHOL, TG, HDL-C, LDL-C, hsCRP levels in ZDF rats were increased gradually (P<0.01). At 16 week, RBP, mALB, TRF contents in ZDF rats were significantly higher than ZL rats (P<0.01). Glomeruli, retina, myocardium, brain tissue showed varying degrees of microvascular disease, vascular basement membrane of the glomerular part appeared thicken, retinal endothelial cells were increased, vascular wall was thicken, and the heart showed a small amount of myocardial fibrosis, hippocampal neurons appeared abnormal. Conclusions ZDF rats showed early microvascular complications of T2DM at16 week, among which T2DM early nephropathy, retinopathy, cardiomyopathy become more prominent, cerebral neuron appeared slight lesion.

Abstract:Objective To establish a swimming endurance system, excavating new indexes evaluating the swim capacity in mice and detecting the effects of HONGJINGTIAN (HJT) capsule on the swimming capacity in mice. Methods Combining video capture technology with image analysis technology into the Exhaustive swimming test, this system would automatically collect and objectively analyze the data when the swimming behavior perform synchronously in mice. A series of indexes would be calculated, such as Exhaustive time, First sink time, Total sink time, Sink frequency, Continuous sink time. Furthermore, the classical anti-fatigue herb named HJT will use to detect the stability and reliability of this swimming endurance system. Results The data showed high correlation between the computer and manual record (r>0.95), indicating high stability and reliability of the swimming endurance system. The continuous sink time (CST) have been proved out high correlation with Exhaustive time(ET) (r=0.974), while the correlation of others indexes less than 0.9. The mice treated with HJT by intragastric administration had showed significantly increase in the First sink time (FST) and the Continuous sink time (CST) compared with the control group. Conclusions The results showed that the system could assess swimming capacity objectively, precisely and sensitively in mice. In addition, this system realized integrating video capture, data analysis and data output together automatically. Besides, a new index-CST was found to be high correlation with ET. This system turn out to be high automatically and intelligently, and provide a reliable system to screen the potential anti-fatigue drugs.

Abstract:Objective To investigate the effects of diflubenzuron on the blood system in rats. Methods A total of 80 rats were randomly divided into three diflubenzuron treated groups and one negative control group. Different doses of diflubenzuron at 0, 25, 500 and 10000 mg/kg were administered continuously for 90 days to the rats by the way of feeding. Animals were observed for clinical signs of toxicity, food consumption and body weight. Haematological parameters, clinical chemistry parameters, absolute and relative organ weights and histopathological changes were examined at the end of the study. Result During the period of the study, the food and water consumption were all normal at all dose groups and no obvious toxicity symptoms appeared. For haematological parameters, the main changes were the decrease of red blood cell count(RBC), haemoglobin(HGB) and haematocrit(HCT) in the medium and high dose groups and the increase of mean corpuscular volume(MCV) and erythrocyte hemoglobin distribution width(RDW) in the high dose group(P<0.05 or 0.01). For clinical chemistry parameters, the main change was the increase of total bilirubin(TBIL) in the high dose group of male rats(P<0.05). The absolute and relative spleen weights increased obviously in the female and male rats of the high dose group(P<0.01). The absolute and relative liver weights increased obviously in the female rats of the high dose group(P<0.01). The spleens were larger, harder and darker than normal in all of the rats of high dose group and part of the rats of medium dose group. For the result of histopathology examination of spleen, the main changes were a lot of red cells in the medullary sinus and haemosiderin in the medulla. Conclusion Diflubenzuron had toxic effect on the blood system of the rats. The spleen was intumesce and chronic hemolytic anemia may be occurred for long-term exposure.

Abstract:Objective To observe the effects of Wuling powder and Hypericum perforatum L. on changes of behavior, the hypothalamus-pituitary-adrenal (HPA) axis function, and monoamine neurotransmitters concentration of hippocampus on chronic unpredictable stress model in rats. Methods Depression model was induced by chronic unpredictable stress combining with solitary method. The stimulus lasted for 4 weeks. During the stress, the rats were intragastric given with the decoction of Wuling powder and Hypericum perforatum L. (490 mg/kg, 245 mg/kg, 123 mg/kg) and Fluoxetine hydrochloride (2.4 mg/kg) for 4 weeks. At the end of the experiment, sucrose consumption and Morris Water Maze experiment monitor the behavior of rats, such as sucrose consumption, escape latency. Moreover, the levels of the monoamine neurotransmitters in the hippocampus were analyzed by high performance liquid chromatography-electrochemistry and the content of CORT, ACTH, CRH in plasma were detected by Radioimmunoassay. Results In Morris Water Maze test, the escape latency of rats which were consecutively administrated with Wuling powder and Hypericum perforatum L. for 4 weeks decreased obviously; the total time and distance in target quadrant remarkably increased (P < 0.05). In sucrose consumption experiment, the percentage in sugar water consumption with treatment of Wuling powder and Hypericum perforatum L. increased significantly (P < 0.01). Meanwhile, the content of norepinephrine (NE), dopamine (DA), and 5-hydroxytryptamine (5-HT) in the hippocampus were significantly increased in rats of Wuling powder and Hypericum perforatum L. and the concentrations of CORT, ACTH, CRH in plasma of Wuling powder and Hypericum perforatum L. group rats were decreased obviously (P <0.05). Conclusions The disturbances of cognition, learning and memory function of rats induced by chronic unpredictable stress combining with solitary could alleviated by Wuling powder and Hypericum perforatum L., and the pharmacological effects may related to modulation of monoaminergic neurotransmitters in the hippocampus and the content of CORT, ACTH, CRH in plasma.

Abstract:Objective To investigate effects of post-treatment on stability of storage solution/thermostability, anti-calcification, the biomechanics and fibred structures of tannic acid fixed bovine jugular vein(BJV). Methods Fresh bovine jugular vein were treated with glutaraldehyde (Glut group), tannic acid (TA group), and post-treatment on TA-fixed (Study group). Calcium level was measured by implanting BJVs subcutaneously into the rats. The shrinkage temperature, tensile strength,elongation at break and histological changes were evaluated as well. Results Compared with Glut group,all results were better in Study group (P <0.05). Compared with TA group, in the Study group, color of storage solution did not change, shrinkage temperature and 60-days' calcium level decreased, 21-days' calcium level increased (P <0.01); biomechanical properties were not statistics difference between TA group and Study group(P > 0.05), and collagen fibers and elastic fibers were maintained well in TA group and Study group. Conclusion Post-treatment on TA-fixed Bovine Jugular Vein can improve stability of storage solution and BJVs elasticity, and do not change anti-calcification, biomechanical properties and fibred structures of TA-fixed BJVs.