Abstract:ObjectiveTo set up a conscious model of vascular headache by electrical stimulation of the superior sagittal sinus and to verify its reliability and feasibility, we observed the behavioral changes of conscious rats which were administrated two classical analgesic drugs that acetaminophen and ibuprofen. Methods30 male SD rats were randomly divided into three groups:control group (saline group), acetaminophen group and ibuprofen group. Each group was given electrical stimulation 5 minutes before and 40 minutes after administrated. The number of head- flicks and excessive grooming times were observed as the main parameters. ResultsThere were significant decrease on head- flicks and excessive grooming between the control group and the administration-group (P<0.05), while no significant difference between the acetaminophen group and the ibuprofen group. ConclusionParacetamol and ibuprofen can significantly relieved the symptoms of the headache in this models by administrated. This conscious rat headache models was practical and reliable model.

Abstract:Objective To establish proper model of human breast cancer in nude mice and study their biological features. Methods MDA-MB-231 and SK-BR-3 cells(ER-) were implanted into the left axilla of 10 nude mice. Tumor growth was observed, and the tumor-bearing mice were killed on day 42. The tumor masses were removed, and pathologically examined by the microscope. ResultsThe MDA-MB-231 tumor tubercles could be formed on day 5. The tumor-take rate was 90%(9/10),and average volume and weight of the tumor were (426.6±333.8)mm³ and (0.417±0.276) g reparatively. The SK-BR-3 tumor tubercles could be formed on day 11. The tumor-take rate was 80%(8/10),and average volume and weight of the tumor were (357.5±246.4)mm³ and (0.325±0.167) g reparatively. All of the specimens were infiltrating ductal carcinoma. Conclusion We succeeded in establishing two human breast cancer model in nude mice, which reserved certain biological features of human breast cancer, therefore it may be an available model for further research of breast cancer.

Abstract:Objective To study the role of Gadd45a gene in mouse hematopoietic stem cell function. Methods Sorting mouse bone marrow hematopoietic stem cells by flow cytometry ,which aim to culture monoclonal; Competitive bone marrow transplantation; The survival curves after irradiation. ResultsGadd45a gene deletion improves hematopoietic stem cell colony forming ability, but hematopoietic reconstitution capacity and survival situation after8.5Gy irradiation is no difference. Conclusion Gadd45a gene plays an important role on mouse hematopoietic stem cell function.

Abstract:Objective To observe the morphology change of human umbilical cord mesenchymal stem cell (hUC-MSC) cultured on polypyrrole (PPy) under the conditions of different styles of electric stimuli. MethodhUC-MSC were identified by flow cytometry and inverted phase contrast microscope. Passage 3 to 6 hUC-MSC were co-cultured with PPy. Current-controlled and voltage-controlled electric stimuli (ES) were applied to the cells. Cells cultured in αMEM with 10% fetal bovine serum and cells cultured on PPy without ES were used as control. Scan electron microscope (SEM) and inverted phase contrast microscope were used to observe the morphology change of hUC-MSC. ResultThe effect of ES on the adhesion and morphology change of hUC-MSC was evident and similar in different groups. Conclusion ES has distinct effect on hUC-MSC adhesion and morphology and can be used as a method of cell regulation. More research are needed.

Abstract:Objective To investigate the feasibility of bone marrow mesenchymal stem cell (hBMSC) transplantation combined with hyaluronic acid poly-l-lysine (HA-PLL) to treat spinal cord injury. Methods hBMSC were extracted from human bone marrow and were cultured and expanded in vitro. The spinal cord of rat was hemi-transected, hBMSC, HA-PLL or hBMSC plus HA-PLL were transplanted alternatively after the injury. The motor function was assayed at different time points. Eight weeks later rats were sacrificed, the growth of axons and micro-blood vessels of different groups were checked, the therapeutic effect of cells, materials and the combination were assayed. ResultsThe progression of rat locomotor function examined by BBB score showed that both groups of hBMSC transplant and hBMSC combined with HA-PLL have better recovery than hemi-transection group and HA-PLL group. Electron microscopy showed combination of hBMSC and HA-PLL can promote axonal and vascular growth. ConclusionshBMSC can promote neurological function recovery. Combination of stem cells and HA-PLL may get better effect. Based on these results, we deduced that the mechanism included the framework supporting effect of materials, the neurotrophic and angiogenesis effect of hBMSC.

Abstract:Objective To establish a haemorrhoids model with the method of acid burns on rats''skin, and to study the pharmacological effects of Tongbianhuazhi Keli on this model. Method10mol /L hydrochloric acid burned the scratched skin around rats anus and the anal medial mucosa. Tongbianhuazhi Keli was orally administered to the rats for 10d. Then score the skin with macroscopic observation and light microscope. ResultPathological results showed that the haemorrhoids model was established successfully. And the pathologic changes on hydrochloric acid burned rats skin can be effectively restored by oral application of large and middle dose of Tongbianhuazhi Keli. Conclusion The haemo haemorrhoids model with the method of acid burns on rats skin was successful; Tongbianhuazhi Keli was effective to restore the rats skin of haemorrhoids model.

Abstract:ObjectiveTo establish heart-specific (pro)renin receptor ［(p)RR］ transgenic mice and investigate its regulation effect on the process of cardiomyopathy. Methods The transgenic vector was constructed by inserting the human (p)RR cDNA sequence into the down-stream of α-MHC promoter. The transgenic mice were generated by the method of microinjection. The genotype of transgenic lines was identified by PCR, and the expression levels of the (p)RR gene were detected by Western Blot and immunohistochemistry. The cardiac geometry and function were detected by echocardiography. Subcellular localization of (p)RR was performed by double immunofluorescence confocal staining. At last, we detected the expression level of ATP2A2, NCX1 and cTnT via Western Blot. ResultsThe heart-specific (p)RR transgenic mice were established and one high-level expression line was identified. Endogenous and transgenic (p)RR were located in the Golgi body and endoplasmic reticulum. Overexpression (p)RR with heart-specific resulted in enhancing contractile properties. Compared with the wild type mice, transgenic mice left ventricular systolic diameter decreased by 9% (P 〈0.05, n=18), left ventricular end-systolic volume reduced by 23% (P 〈0.05, n=18), ejection fraction increased by 14% (P 〈0.01, n=18) and fraction shortening elevated by 20% (P 〈0.01, n=18). The expression level of the proteins involving in Ca²⁺ and contractile properties were altered, SRECA and NCX1 decreasing, while cTnT increasing. Conclusion Over-expression of (p)RR in the heart of the transgenic mice leaded to significantly enhanced cardiac contractile properties., and while regulated the expression of SRECA, NCX1 and cTnT, directly or indirectly. These results indicated (p)RR participated in influencing cardiac function via regulating the Ca²⁺ of the myocardium.

Abstract:Objective To establish the mouse model for application in the study of pathogenesis of EV71 infection and evaluation of vaccines or drugs. Methods A mouse-adapted strain MP10 was generated by a series of passages of the parental EV71 strain in the skeletal muscle of mice. And then the symptoms, virus replication and pathological changes of tissues of infected-mice were monitored. ResultsCompared to the parental strain, MP10 showed enhanced infectivity in RD cells and virulence in mice, and MP10 infection caused paralysis and death in 10-day-old mice. Skeletal muscle was the main site for virus replication and showed extensive necrotizing myositis, whereas no lesions were observed in neurological tissues. The muscles associated with respirometric activity showed severe necrosis, which might have caused the infected mice to experience difficulty in breath and was thought to be the cause of death. Conclusion The 10-day-old mouse model for lethal EV71 infection was established and it would be useful for vaccine and drug evaluation.

Abstract:ObjectiveTo investigate the influence of Exo-1 gene deletion on the aging of the hematopoietic stem cell environmental defect of telomerase-deficient mice. Methods Recipients were the 3^rd generation telomerase knockout mice (G3 Terc-/-) or the 3^rd generation telomerase and Exo-1 gene double knockout mice (G3Terc-/-Exo-1-/-). Bone marrow cells were isolated from CD45.1 mice, and then injected into recipients at 2 month of age. Recipient mice were sacrificed at 9 month of age, the bone marrows, spleen, thymus and peripheral blood were collected and analyzed by flow cytometer. ResultsPercentage of B220⁺ cells increased in the wildtype donor derived bone marrow, spleen and peripheral blood cells in G3 Terc-/-Exo-1-/- double knockout recipient mice compared to those in G3Terc-/- recipient mice. Conclusion Exo-1 gene deletion rescued the hematopoietic stem cell environmental defect, and reversed the abnormal B cell development of hematopoietic stem cells.

Abstract:Objective To identify genotypes and potential routes of dissemination of P.aeruginosa isolates in Beijing using Multilocus-locus variable-number tandem-repeat(VNTR) analysis (MLVA). Methods One hundred and forty one experimental animals and facilities isolates of P.aeruginosa were detected by selected 13 VNTRs loci. Then we draw the phylogenetic tree and minimum spanning tree(MST) using BioNumerics software according to fingerprint clustering analysis. Results All isolates of P. aeruginosa was divided into three major genetic Clusters, 56 genotypes. The proportion of A cluster is 82.3%, B cluster 12.8%, C cluster 5.0%. The Simpson’s index of diversity is 0.763. The isolates have distant homology in the neighboring facilities. ConclusionsMLVA displayed excellent discriminatory power for P.aeruginosa. MLVA can effectively track P.aeruginosa source. The P. aeruginosa isolates of experimental animals have rich polymorphism genotypes and no regional homology in Beijing.

Abstract:Objective To analyze some potential factors which might affect the active systemic anaphylaxis (ASA) test on guinea pig. Methods In GLP conditions, guinea pigs from 300 to 400 grams were used as experimental subjects. By using the materials including bovine serum albumin and saline which are recommended for the control subjects by the guidelines, some experimental design factors such as sensitizing dose, challenge dose, challenge interval and challenge injection route, and the frequency of challenge were studied during the systemic anaphylaxis in guinea pigs. ResultsIn a certain dose range, the incidence of allergic reactions in guinea pigs was not related with systemic sensitizing dose, challenge dose and challenge injection approach, but the severity of anaphylaxis symptoms has an obvious relationship with sensitizing dose, challenge dose and challenge injection route. Conclusion Study on the drug-induced anaphylaxis should refer to the guidelines and pay more attention to experimental design dose and the route of administration.

Abstract:Objective To investigate the cause of formation of herniation found in adult male humanized mice and the effect of that herniation on humanized mice. Methods Construction humanized mice with the method of injection human hematopoietic stem cells into murine blastocysts. The phenotypic characteristics and pathological mechanism of herniation, the effects of herniation on behavior and physiological reaction of humanized mice were analyzed. Results2 months of age or older male humanized mice developed direct inguinal hernias. The loss of dense connective tissue may be the main reason of the inguinal hernias. The motor coordination and endurance of humanized mice decreased as compared with wild-type mice, but the reproductive system of male humanized mice have no difference from the wild-type mice. ConclusionsAdult male humanized mice suffer from inguinal hernias, the formation mechanism of which should be further researched.

Abstract:ObjectiveTo investigate the safety and efficacy of immediately coronary optical coherence tomography (OCT) examination after stent implantation in the mini pig model. Methods Twenty-two health Chinese mini pigs were given percutaneous coronary stent implantation in right coronary artery. Stent strut malapposition was evaluated by OCT analysis. The duration and complication of OCT examination were recorded at the same time. ResultsOCT examination was performed on 25 stents and failed in 3 cases (1 pig had guiding wire fractured inside stent and 2 had coronary artery spasm and failed to deliver the occlusion balloon). Side effect contained 5 cases of temporarily ST segment elevation and 3 cases of ventricular fibrillation (2 pigs survived after successful electrical defibrillation and 1 dead). The mean duration of operation and OCT examination was 49.7±12.9 minutes and 17.9±4.9 minutes respectively. OCT detected 4514 stent struts and discovered 66 malapposition struts. And the rate of malapposition strut immediately after stent implantation was 1.46%. ConclusionOCT examination could be applied on mini pig stent implantation model with high quality and safety and was effective enough to evaluate the strut malapposition immediately after stent implantation.

Abstract:ObjectiveIn order to improve the method of setting up the model of rat cryptorchidism mode, we explored a new method and also examined its stability. MethodTwenty-eight male Wistar rats were randomly divided into control (Ctrl) and model (Modl) group. Each rat was kept in a single cage. The Ctrl group can move freely during the experiment. The Modl group was tail-suspended for 3 weeks, and then reloaded for 8 weeks. Rats were sacrificed at the end of 3^rd week and 11^th week. The testis and epididymis were removed, weighted, then stained by Hematoxylin and Eosin Stainin, and examined under light microscopy. ResultsAfter 3-week tail-suspending, all rats’ testis slipped into enterocoelia. Compared with Ctrl Group, the weight of testis and epididymis were significantly decreased (P<0.01). After 8 weeks recovery, the weight of testis and epididymis did not increase to normal (P<0.01). With morphological examination, we found that after 3 weeks tail-suspending, seminiferous tubules were completely disorder and the spermatogonium were disappeared in the cauda epididymidis. The pathological changes were not improved(P<0.01)at the end of 11^th week. Conclusion3-week tail-suspending can simulate the pathological changes of cryptorchidism syndrome. It is much easier to be done than the traditional one. Moreover, this method has less injury on the rat. Therefore, it can be used to set up a new model of cryptorchidism syndrome.

Abstract:Objective In order to prepare immunoglobulin G (IgG) secondary antibody of non-human primates such as Macaca fascicularis and Macaca mulatta and establish a quick serological method to diagnose their diseases. Methods Macaca fascicularis IgG was isolated and purified from Macaca fascicularis serum by precipitated use saturated ammonium sulfate and affinity chromatography technology with Agarose-Protein G. After identified by SDS-PAGE electrophoresis murine monoclonal antibody(McAb)were produced by conventional immune method and hybridoma technology.Monoclonal antibody was produced when C57BL/6 mice was immunized three times and its spleen cells were fused with SP2/0-Ag14 myeloma cells. Indirect ELISA and Western blot methods were used for identification. Result5 strains of myeloma cells acquired which were named 2B₆, 2B₇, 3B₂, 5E₄, 2D₉.The antibodies they secret can all react well with both IgG and their serum of Macaca fascicularis and Macaca mulatta. There are no cross reaction with other species such as east-northern tigers and dogs. Conclusion The 5 strains of monoclonal antibodies can secrete antibody chronically and steadily. The study can be valuable in establishing a quick serological method in diagnosing the infectious disease.

Abstract:The revised Directive for the protection of laboratory animals 2010/63/EU will be put into practice from January 2013. There are some articles amendment for animal welfare and 3Rs in new Directive comparative with its predecessor, which represent the new concepts for using animal testing and administration. The changes process of EU Directive and adjust for adapt to social status and scientific progress, which is worthy to reference for Chinese labanimal standardized management and assimilated to mainstream criterion. To understand the articles contents is useful to laboratory animal industries participate in the international competition, contrast research and provide specialized service in foreign trade.