Abstract:ObjectiveTo observe the changes of pancreatic islet cells in experimental macaques diabetes models. MethodsWe used streptozotocin (STZ) (30 mg/kg body weight, intravenously, 3 days) to five macaques. The animal was sacrificed when it was going to die. The changes of A, B, D, PP cells in the pancreatioc islets were observed with immunohistochemistry staining. Image analysis and statistical analysis of the histological changes were carried out. Results The quantity of A cells was significantly decreased than that in the control group (P<0.01). The positive immunohistochemical reaction of A cells was significantly increased compared with that of the control group (P<0.01). The positive PP cells were increased than that in the control group (P<0.05). There were no significant differences in D cells between the model and control groups. ConcusionThe number of pancreatic islet cells and changes of hormonal expression in experimental macaques models of diabetes mellitus is similar to that in diabetic human subjects.

Abstract:ObjectiveTo evaluate the domestic ELISA kit for detection of mouse virus antibodies.MethodsDomestic and imported ELISA kits were choosen to detect five mouse virus antibodies , including lymphocytic choriomeningitis virus (LCMV), hepatitis virus (MHV), Sendai virus (SV), adenovirus (MAV), and parvovirus (MPV). The sensitivity, specificity, precision, stability, and reliability of the five kits were compared. ResultThere were significant differences in sensitivity of the same type domestic and imported kits, varying from at least 2 times (P<0.05) up to 16 times (P<0.01), and there were no cross reaction with the other four types of viruses. Precision test showed that the batch average coefficient of variation of the five kits was less than 10%. Stability test showed that the relative deviation of the five kits was less than 25%. 36 known mouse serum samples were tested with both the domestic and imported kits, respectively, and showed a coincidence rate of 100% for detection of LCMV, MHV, SV, and MPV. The domestic kit showed a coincidence rate of 86.1% in test for MAV, and that of the imported kit was 100%, with a very significant difference between them (P<0.01). ConclusionsExcept that the domestic MAV kit shows a lower sensitivity and credibility than that obtained with the imported kits, the domestic and imported LCMV, MHV, SV, MPV kits showed similar sensitivity, specificity, precision, stability and reliability.

Abstract:ObjectiveTo study the total amount of expression of RT1A, RT1B and RT1D gene antigens in different rat strains, their amount of expression in CD4+ T cells and CD8+ T cells, and provide support data for immunology researches.MethodsVenous blood was collected from the rats of BN, Lewis, F344, SHR strains. Lymphocytes were isolated, and reacted with fluorescent marker of monoclonal antibody. The amount of gene antibodies was assessed by flow cytometry. ResultsThe RT1A, RT1B and RT1D gene antigen expressions were different in different rat strains. Among them, the expression of RT1A antigen was highest in F344 rats, and expression of RT1B and RT1D gene antigens was highest in BN rats. The amount of expression of RT1A, RT1B and RT1D gene antigens on CD4 + and CD8+ T cells were also different. Among the studied rat strains, the expression of RT1A antigen on CD4+ T cells was highest in Lewis rats, and the expression of RT1B and RT1D gene antigens on the CD4+ T cells was highest in BN rats.. The expression of RT1A antigen on CD8+ T cells was highest in F344 rats, and the expression of RT1B and RT1D gene antigens on CD8+ T cells was highest in BN rats. A higher expression of RT1A, RT1B and RT1D gene antigens was found in the male rats than in female rats of the same strain. ConclusionsThere are significant differences between the total amounts of expression of RT1A, RT1B and RT1D gene antigens in different rat strains, and also significant differences between the amounts of their expression on CD4+ and CD8+ T cells in these rat strains. Our results indicate that it is important to choose appropriate rat strain to provide reliable and proper results in immunological researches.

Abstract:ObjectiveTo investigate the impact of cigarette smoke exposure on the expression of CCR6 in airway of asthmatic rats. MethodsForty male Wistar rats were randomly divided into four groups with 10 rats in each group:control group, cigarette smoke exposure group, aerosolized ovalbumin (OVA) exposure group and OVA combined with cigarette smoke exposure group. Rats were exposed to either aerosolized OVA , tobacco smoke, or both tobacco smoke and OVA. White blood cells in BALF were quantitatively analyzed and classified. The expressions of CCR6 mRNA and protein in the airway were detected by immunohistochemistry and real-time RT-PCR. Results① The number of white blood cells, the percentage of eosinophils and neutrophils to total cells in BALF in the aerosolized OVA exposure group (69.0±3.5,4.1±1.0,8.9±2.0) and OVA combined with smoke exposure group (86.7±5.2,2.2±1.0,9.0±2.8) were higher than that in the control group (10.1±3.8,1.3±0.7,2.2±1.1) and smoke exposure group (47.7±6.8,0.5±0.3,2.7±1.4) (all P<0.05). The number of white blood cells, the percentage of neutrophils to total white cells in BALF in the OVA combined with smoke exposure group were significantly higher than that in the aerosolized OVA exposure (P<0.05). The percentage of eosinophils to total white cells in BALF in the OVA combined with smoke exposure group was lower than that in the aerosolized OVA exposure group (P<0.05). ② The expression of CCR6 mRNA and protein in the aerosolized OVA exposure group (8.15±0.88;0.452±0.013) and OVA combined with cigarette smoke exposure group (15.16±0.7,0.531±0.024) were significantly higher than that in the control group (1.01±0.2,0.299±0.027) and the smoke exposure group (5.55±0.4,0.442±0.018) (all P<0.01). The expressions of CCR6 mRNA and protein in the OVA combined with smoke exposure group were significantly higher than that in the aerosolized OVA exposure group (all P<0.01). ConclusionsThe data of this study suggest that cigarette smoke can aggravate chronic airway inflammation by increasing the expression levels of CCR6 mRNA and protein in asthmatic rats.

Abstract:ObjectiveTo establish a RT-PCR detection method for Sendai virus and to investigate the infection status in Cricetulus migratorius. MethodsTo design PCR primers according to the Sendai virus(gi:9627219)genome in NCBI and establish a RT-PCR detection method using the Sendai virus strain from our department. To screen the Sendai virus infection in sixty Cricetulus migratorius lung samples by the established RT-PCR procedure. ResultsThe RT-PCR method established was more sensitive and specific:there was a 197 bp single band when SV was used as template and no any band occurred when SV5, CDV, PVM, Reo3 and MuV were used as templates. Sequencing results revealed that the 197 bp nucleotides showed 98% coincidence with the SV sequence published by NCBI. The assay could detect as low as 96.8 ng/mL SV cDNA. Results showed that the infection rate of Sendai virus was 3.33%(2/60)in Cricetulus migratorius. ConclusionsThe established RT-PCR detection technique is sensitive, specific and rapid, and can be used to detect Sendai virus routinely in laboratory rodent animals. The positive results of Sendai virus infection show that Sendai virus infection in Cricetulus migratorius can not be ignored.

Abstract:ObjectiveTo diagnose the subcutaneous tumors in the NIH mice in bred in the laboratory animal center in our company.MethodsTo statistically analyze the morbidity, body weight and parturition times of the NIH mice bred in our animal center. Tumor samples were taken for pathological examination. ResultsOne hundred and fifty-four subcutaneous tumors were found among 11711 mice. The pathological diagnosis of the subcutaneous tumors was fibroadenoma, and the incidence rate was 1.32%. ConclusionsThe tumor of NIH mice bred in our company laboratory animal center is fibroadenoma. The etiology of this tumor needs to be further investigated.

Abstract:ObjectiveTo establish an upper thoracic epidural anesthesia model in rats. MethodsA total of 120 rats were used in this study. Rats were fixed in a prone position, and an epidural catheter was implanted into the spinal epidural space through the T4-5 joint in cephalad direction. At 48 hours after catheterization, 100 μL/kg methylene blue was injected into the epidural cavity in rats to evaluate whether the catheterization was successful or not, and to identify the location and distribution of the drug solution. ResultsThe thoracic epidural anesthesia model was successfully performed in 85.8% of the rats. The infection rate of the epidural cavity was 0, and the rate of catheter prolapse was 5% after 48 hours. ConclusionsT4-5 cephalad catheterization is feasible to provide a stable upper thoracic epidural anesthesia model in rats, with apparent advantages such as mild traumatic injury, good stability, high success rate, lower cost, and less time consuming. 

Abstract:ObjectiveTo compare the efficacy of Dixon and Mood up-and-down method and Karber method in determining tetanus toxin LD₅₀ in mice. Methods Tetanus toxin in a dose of 0.5 mL at concentration of 0.9,0.1,0.5,0.0,0.267 or 0.256 ng/mL was subcutaneously injected into NIH mice. The LD₅₀ of tetanus toxin was determined by Dixon and Mood up-and-down procedure and Karber test. ResultsThe LD₅₀ of tetanus toxin was 4.97 ng/kg body weight measured by Karber method, and 4.93 ng/Kg body weight by the up-and-down procedure. Conclusions Both Dixon and Mood up-and-down procedure and Karber test can achieve the same LD₅₀ of tetanus toxin, but the up-and-down procedure is more simple and convenient, and 8 animals are enough for this test.

Abstract:Objective To analyze the polymorphism of GH gene in Tibet miniature pig and Guangxi Bama miniature pig by PCR-RFLP. Methods The polymorphism of the -119bp-+486 bp fragment (growth hormone gene) of 108 Tibet miniature pigs and 132 Guangxi Bama miniature pigs was detected by PCR-ApaI-RFLP and PCR-Hin6I-RFLP. Results (1) The PCR products were cut by ApaI, and produced two alleles:A (449 bp + 101 bp + 55 bp) and B (316 bp + 133 bp + 101 bp + 55 bp)。The frequency of allele A was higher than that of allele B. The frequency of the AA genotype was higher than that of the AB and BB genotypes. (2) The PCR products were cut by Hin6I, and produced four alleles:G1 (605 bp), G2 (498 bp+107 bp), G3 (449 bp+156 bp) and G4 (449 bp+107 bp+49 bp). The allele frequency of G4 was higher than that of G1, G2 and G3. The frequency of allele G1s was the lowest. Both Tibet miniature pig and Guangxi Bama miniature pig exhibited moderate polymorphism. The frequencies of G2G3, G2G4, G3G4 and G4G4 genotypes were higher than others. (3) There was no significant difference between the expressions of the allele in the Tibet miniature pig and Guangxi Bama miniature pig revealed by PCR-RFLPs. ConclusionsThe frequency of allele A of GH gene in Chinese miniature pig species, such as Tibet miniature pig and Guangxi Bama miniature pig, are rather high.

Abstract:ObjectiveTo establish a multiplex PCR procedure for simultaneous detection of H. hepaticus, H. bilis, and H. rodentium, and to preliminarily verify its application value.Methods Three pairs of specific primers for the multiplex PCR procedure were designed according to published H. hepaticus, H. bilis, and H. rodentium 16S rRNA gene sequences, and the reaction conditions of the multiplex PCR were optimized. ResultsThe results showed that all the three pairs of primers could be used to specifically amplify their target bands of 417 bp, 364 bp and 324 bp. The optimal annealing temperature was 52℃, the Mg²⁺ concentration was 2.0 mM, the dNTP concentration was 200 μΜ, and the concentration of primers was 0.625 μM. The sensitivity of the multiplex PCR for H. hepaticus, H. bilis, H. rodentium was 10 fg. CouclusionsA sensitive, specific, efficient multiplex PCR procedure has been established in this study, providing a good foundation for the detection of H. hepaticus, H. bilis, and H. rodentium.

Abstract:ObjectiveTo establish a PCR detection method for simian adenovirus (SAdV) and study the SAdV infection status in monkeys. Methods To design a primer based on SAdV sequences and optimize the PCR experiment, test the method, to detect the virus in monkeys and vaccines, and sequenceing and constructing the phylogeny trees. ResultsA PCR detection method for SAdV was established, which could distinguish SAdV from MAD, ICH, CELO and was sensitive to 47.9 pg/mL viral DNA. The results of this study showed a positive rate of 49.2% and a large genetic diversity of simian adenoviruses in Chinese primate colony. Most of them belong to G subgroup and twig of SAdV-49. ConclusionsThis PCR detection method established in our laboratory has a good accuracy. We found a high prevalence of SAdV in Chinese primate colony. It is necessary to enhance detection and control for SAdV in monkeys and relevant biological products, and avoid potential risk of human infection.

Abstract:ObjectiveTo observe and analyze the neuroimaging characteristics of the brain tissue of Alzheimer''s disease (AD) transgenic mice models. MethodsThe in vivo microstructural changes in brain tissue of AD transgenic mouse models and wide type mice (1-, 3-, 5-, 7-, 9- and 11-month old) were compared with (7.0 T) high-field intensity magnetic resonance imaging. The changes in the cerebral parietal cortex and hippocampus of AD transgenic mice at different ages were quantitatively analyzed by transversal relaxation time (T₂), apparent diffusion coefficient (ADC) and fractional anisotropy (FA). ResultsScattered low signal zones were observed in the cerebral parietal cortex and hippocampus of AD transgenic mice aged 9 months and above. The T₂ relaxation time was decreased in the parietal cortex and hippocampus of AD transgenic mice aged 1 to 9 months, whereas there was no significant difference between the AD transgenic mice and wide type mice (P≥0.05). The ADC value in the parietal cortex and hippocampus had a decreasing tendency in the AD transgenic mice aged 7 to 11 months. A reduction of FA value was detected in the APP/PS1 mice aged 5 to 11 months, compared with that of wide type mice (P≤0.05). ConclusionsHigh-field intensity MRI can reveal changes in the cerebral parietal cortex and hippocampus of AD transgenic mice by changes of FA values, suggesting that FA value could be a useful reference parameter in early diagnosis of Alzheimer''s disease. 

Abstract:ObjectiveTo observe the changes of blood biochemistry and blood gas and electrolytes in sub-health model rats caused by different factors. Method60 SPF male Wistar rats were randomly divided into 5 groups (n = 12):multifactor group (MF), warm swimming group (WS), sleep deficiency group (SD), purely constraint group (PC) and control group (C). We used the way of single stimulation such as warm swimming, sleep deficiency or complex stimulation to establish the sub-health models. Six rats were sacrificed 5 days after modeling. The others were sacrificed 3 days after recovery breeding. The serum ALT, AST, TP, ALB, BUN, CREA, GLU, TC, TG, CK, LDH levels and blood gas and electrolytes were detected. Result(1) Compared with the control group, the serum BUN and TG in the MF group were significantly lower (P<0.05, P<0.01) 5 days after modeling, while CREA, AST, CK, CK/AST and LDH levels were significantly increased (P<0.01). The levels of TC, CK, CK/AST and LDH in the WS group were also significantly increased (P<0.01) and GLU concentration in all the groups was significantly increased (P<0.05). After recovery breeding for 3 days, the levels of CREA, AST, CK, CK/AST and LDH in MF group were significantly higher than those in the control group (P<0.01), while the CK activity in the SD group was significantly higher than that in the control group (P<0.05). (2) Compared with the control group, PCO₂, Hct and Hb in the MF, SD and PC groups were significantly increased at 5 days after modeling (P<0.05, P<0.01), while Hct and Hb in the WS group were significantly increased and Na⁺ concentration decreased (P<0.01). PO₂ and SO₂% in the MF group were significantly decreased and K+ concentration was increased (P<0.05). The HCO-₃ concentration of all the groups was significantly increased. After recovery breeding for 3 days, except for pH and PCO₂ were significantly decreased in all model groups (P<0.05, P<0.01), PCO₂ in the MF, SD and PC groups was significantly increased and SO₂% was decreased (P<0.05, P<0.01), Na⁺ concentration in the MF and WS groups was still lower than that in the control group, and K⁺ and Mg⁺⁺ concentrations in the SD group were significantly higher than those in the control group (P<0.05, P<0.01). ConclusionsBlood biochemistry, blood gas and electrolytes indexes are differently changed in sub-health rats induced by different factors, thus these indexes could be used to evaluate the changes of blood microenvironment in the sub-health state. 

Abstract:ObjectiveTo study the influence of enriched environment on the growth and behavior of guinea pigs. MethodsThe breeding environment of guinea pigs was enriched by adding tube-shaped mould and trapezium-shaped mould. Thirty-two healthy guinea pigs were randomly divided into 4 groups, with 8 animals in each group:normal control, tube mould, trapezium mould, tube and trapezium mould groups. The body weight of guinea pigs was recorded every week for 4 weeks, and the behavior changes of the guinea pigs were observed. ResultsThe growth of guinea pigs bred in the enriched environment was better than that of the control animals, and the tube mould showed better effect than the trapezium mould. ConclusionsEnriched environment with moulds is beneficial for guinea pig breeding. The tube-shaped mould is more close to a natural cave, may be beneficial for the guinea pigs to hide, and provides for them a better growing and living environment.

Abstract:The facilities for large laboratory animals were constructed under reasonable design and specification according to the national standard(GB14925-2010, laboratory animal- requirements of environment and housing facilities)combining with building structure. All indices of the facilities achieved the related national standard and work well until now. Here we share some experience in the construction of the facilities.