Laboratory Animal Science

The Study of Expression of NF-κB in Ｒats’Liver after Welfare Impaired by Experimental Procedures

2016, 33(06): 1-4.

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Abstract: Objective To study the expression of NF-κB in rats’liver after welfare impaired by experimental procedures． Method Male SD rats，welfare were impaired by experimental procedures． Body weight gain and food consumption were recorded throughout the study ，mＲNA of NF-κB p65 in liver were detected by real-time quantitative PCＲ，and protein expression were detected by Western blotting．Ｒesult Compared with the control group ，the body weight gain and food efficiency ratio ( FEＲ) of experimental group were decreased ( P ＜ 0. 01) ，but mＲNA and protein expression of NF-κB p65 were much higher ，and showed very significant difference ( P ＜ 0. 01) ． Conclusion Welfare impaired caused by the stress of experimental procedures ，the expression of NF-κB in rats’liver are much higher than before． It shows that NF-κB participate in the process of welfare impaired．

Effects of Alpha-Linolenic Acid Tolipid Synthesis Gene in Insulin Ｒesistance HepG2 Cells

2016, 33(06): 5-10.

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Abstract: Objective We investigate the effect of ALA on lipid biosynthesis-related genes in animal and cell IＲ models． Method HepG2 cells were divided into two groups，normal control group ( NC) and palmitic acid group ( PA) in which cells were firstly cultured for 24 h in the medium contained 0. 25 mmol /L palmitic acid． The glucose uptake，cellular TG and TCH level were determined and once there was significant difference in glucose uptake，ALA took 20 percent the place of PA for another 12 h culture． Celluar TCH and TG level were detected and mＲNA and protein expression of genes related to lipid synthesis were examinated by real-time qPCＲ and Western blot．Ｒesult After incubation with 0. 25 mmol /L PA for 24 h，the glucose uptake of PA group was significantly lowered than NC group indicating the IＲ-HepG2 cell model was established characterized by extremely high TCH level． ALA selectively upregulated the protein expression of INSIG-2 and decreased the TG level in IＲHepG2 cells，and there were no statistically significant differences in TCH level and INSIG-1 protein expression． ALA had no significant effect on the abundance of the transcriptionally active ( 65 kDa) form of Srebp-1c while the Fasn protein reduced by 21% after ALA intervention． When compared with PA，ALA reduced 55 － kDa cleaved form of SＲEBP-2 and Hmgcr protein． Conclusion ALA selectively upregulated the protein expression of INSIG-2 in IＲ-HepG2 cells，rather than Insig-1． ALA reduced the abundance of post-translational form of SＲEBP-2 and its target Hmgcr and Fasn by increasing the expression of INSIG-2．

Biodegradable Mesh-like Microporous Balloon Combined with Calcium Phosphate Bone Cement for the Treatment of Fracture: A New Zealand Ｒabbit Study

2016, 33(06): 11-15.

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Abstract: Objective To investigate the degaration of femoral implant materials and blood index changes to identify the therapeutic effect of biodegradable mesh-like microporous balloon and calcium phosphate bone cement for the treatment of bone fracture in New Zealand rabbit． Method The rabbit models were randomized into two groups: the experimental group with biodegradable mesh-like microporous balloon and calcium phosphate bone cement and the control group only with calcium phosphate bone cement． Clinical parameters such as blood cell count were examined and compared with intra-group and extra-group at pre-operation，and 1，3 and 6 months after operation． Moreover，the degradation states of implant and the growth situation of bone in the two groups were compared according to the imaging data．Ｒesult No statistical significance for blood cell counts was found between pre-and post-operation or between two groups． The materials in the two groups had favorable biocompatibility without obvious immunological response． Bone tissue can grow into bone cement gaps，to replace the degradation of bone cement and formation rules of trabecular bone structure． The trabecular structure of experimental group were bulky density，distribution rules than control group． Conclusion The research verified that it has no obvious immune response while putting the degradable mesh porous balloon calcium salt into living bone cement，and is conducive to the restoration of fracture healing in New Zealand rabbit．

Investigation of Blood Ｒoutines and Intestinal Mucosal Pathologies of Dry-heat Stroke Ｒats

2016, 33(06): 16-20.

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Abstract: Objective To investigate the changes of blood routines and intestinal mucosal pathologies of different degree heat stroke rats in dry-heat environment of desert． Method Forty eight male SD rats，SFP grade，were randomly divided into normal control group，light heat stroke group，moderate heat stroke group and severe heat stroke groups． Placed in simulated dry-heat environment of desert in The Simulated Climate Cabin for Special Environment of Northwest of China，temperature of ( 41 ± 0. 5) ℃，humidity of ( 10 ± 2) %ＲH，fasting and water deprivation，for ( 50 ± 5) min，( 100 ± 5) min，( 150 ± 5) min respectively to cause light，moderate，severe heat stroke rat models． Then the rats were deep anesthetized and sacrificed immediately with 3% sodium pentobarbital intraperitoneally，samples of venous blood and ileum and colon tissue were collected to examine blood routines and count pathological histology score．Ｒesult Ｒats with accumulated degree of heat stroke，white blood cell count increased gradually，white blood cell count of SHS group compared with the other groups increased significantly ( P ＜ 0. 05) ; red blood cell count and hematocrit of MHS group compared with the NC group and LHS group increased significantly，SHS group increased significantly，too ( P ＜ 0. 05) ; platelet count of MHS group compared with the NC group and SHS group increased significantly ( P ＜ 0. 05) ． The pathology injury score of ileum and colon were significantly increased with increasing heat degree in MHS group and SHS group ( P ＜ 0. 05) ． The ileum pathology injury score of MHS and SHS groups were higher than that of colon ( P ＜ 0. 05) ． Conclusion White blood cell count，red blood cell count，hematocrit of heat stroke rats，in dry-heat environment of desert，were increased，platelet count showed a trend of firstly increased and then decreased，intestinal mucosa pathological changes with increasing heat degree aggravated． Ileum pathology injury compared with colon is more serious． Our results showed that protection of small intestinal mucosa of heatstroke rats is one of the effective way to prevent heat stroke in dry-heat environment，and platelet count decreased may be one of the key parameters to reflect the severe heat stroke．

Study of Curcumin Neuroprotective Effects on Heatstroke Ｒats in Dry － heat Desert Environment

2016, 33(06): 21-24.

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Abstract: Objective To study the neuroprotective effect of Curcumin on the oxidative stress indexes of heatstroke rats in dry-heat environment of desert． Method The 200 male SPF SD rats，were randomly divided into control group，low-dose Curcumin pretreated group，middle-dose Curcumin pretreated group， high-dose Curcumin pretreated group and solvent group，each group had 40 rats． All the rats were put into the dry heat environment ( temperature of ( 41 ± 1) ℃，humidity 10%) in The Simulated Climate Cabin for Special Environment of Northwest of China． Each group were randomly selected 10 rats to measure oxidative stress indexes-brain malondialdehyda ( MDA) and glutathione( GSH) levels at the time of 0 min，50 min，100 min，and 150 min．Ｒesult There were no significant difference of MDA and GSH levels among the groups at 0 min( P ＞ 0. 05) ． The MDA levels of rat’s brain tissue increased and GSH levels reduced with prolonged in dry heat environment． The MDA and GSH levels of brain tissue of high dose Curcumin pretreatment group compared with the control group and the solvent group， were significantly different ( P ＜ 0. 05) ． Conclusion Curcumin can reduce the MDA levels of brain tissue and increase the GSH level of heatstroke rats in dry heat environment of desert． Curcumin has a protective effect on the heatstroke rats in the dry heat environment of desert．

Study on Protective Effect and Mechanism of Hepatocyte Growth-promoting Factor on ANIT-induced Acute Liver Injury in Mice

2016, 33(06): 25-30.

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Abstract: Objective To study the protective effect and mechanism of hepatocyte growth-promoting factor ( PHGF) on ANIT-induced liver injury in mice and provide theoretical basis for its clinical use． Method Fifty male BALB/c mice were divided randomly into normal group ( control group，n = 10) and ANIT treatment group( n = 40) ． Liver injury model in mice was established by oral administration of ANIT( 105 mg /kg) ，recombinant human hepatocyte growth factor ( rhHGF，1 μg /mice) and 2 doses of PHGF( 10，20 mg /kg) were administrated intraperitoneally 0. 5 h before and 6，12，24，30，36 h after ANIT administration． 48 h after ANIT administration，animals were anesthetized and heart blood，the ALT，AST，ALP levels in serum and SOD level in liver tissue were detected． Furthermore， liver tissues were observed histologically by HE staining．Ｒesult ALT，AST and ALP levels in PHGF and rhHGF group were decreased significantly compared to those of the model group( P ＜ 0. 05) ． SOD level in ANIT group was significantly higher than that in the control mice． However，there is no significant between ANIT and rhHGF， PHGF groups． Through the pathological observation，necrotic area and inflammatory cell infiltration of liver in rhHGF and PHGF group of liver cell necrosis situation was less severe than that of the model group． Conclusion PHGF can protect the ANIT-induced liver injury in mice and the mechanism of action may associated with the inhibition of inflammation

Effect of Celecoxib on the Expression of Iba-1 and GFAP in Epileptic Ｒats Induced by Kainic Acid

2016, 33(06): 31-34.

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Abstract: Objective To study the protective effct of celecoxib on epileptic rats．Method Kainic acid( KA，1 μg /μL，1. 0 μL) was injected into hippocampus to establish epilepsy model in rats and surveyed the behavioristics changes; immunohistochemical analysis was used to detect the Iba-1 and GFAP of hippocampus．Ｒesult Compared with model group，celecoxib group extended the incubation and reduced the largest seizures rank significantly; decreased the expression of Iba-1 and GFAP，attenuated the hippocampus glial activation caused by KA． Conclusion Celecoxib may inhibit the activity of gia cell to alleviate the gial-mediated inflammation，and lighten the degree of epilepsy．

Prokaryotic Expression and Antibody Preparation of Human NEDD8

2016, 33(06): 35-38.

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Abstract: In the study，human nedd8 gene ( HN8) was synthesized and subcloned into pMD19-T simple vector．After sequencing，HN8 gene was transferred into the prokaryotic expression vector，pCold-TF． The final construct is designated as pCold-HN8． The plasmid was transformed into E． coli BL21 ( DE3) competent cells． Induced with Isopropyl β-D-1-thiogalactopyranoside ( IPTG) ，the bacteria were grown in LB broth at 16℃ for 24 hours． The products were sonicated and then purified by His-bind purification kit． The purified protein was used to immunize three rabbits to prepare polyclonal antibody． The results showed that the fused protein were soluble expressed with a size of 64 kDa by SDS-PAGE analysis． Both indirect immunofluorescence and Western blotting assay，it confirmed the polyclonal antibody could recognize the HN8 protein． This antibody provides a useful tool for a further study of the neddylation modification by human NEDD8．

Study on the Difference of Heart Ｒate Ｒeactions about Potential Guide Dogs in Fear Test

2016, 33(06): 39-44.

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Abstract: Objective To explore the heart rate reactions of guide dogs in the fear test，and to provide effective behavioral and physiological basis for early identify of dogs best suitable to guide dog work． Method 28 guide dogs and 32 eliminated dogs which were provided by Chinese Guide Dog Training Centre of Dalian were tested by fear test． The heart rate reactions of 16 guide dogs and 16 eliminated dogs were recorded in the test． The score of “Attention”，“Active Exploration”in fear test，and the heart rate reactions were analyzed．Ｒesult The score of “Attention”and “Active Exploration”of guide dogs were significantly higher than those of eliminated dogs ( “Attention”: in guide dog group was 3. 32 ± 0. 18 and in eliminated dog group was 2. 34 ± 0. 18，P ＜ 0. 01; “Active Exploration”: in guide dog group was 3. 32 ± 0. 24 and in eliminated dog group was 2. 34 ± 0. 18，P ＜ 0. 01 ) ． The heart rate reactions in the fear test were significantly higher than the resting heart rate in both guide dog group and eliminated dog group，respectively ( in guide dog group，heart rate at test: 116. 76 ± 11. 11 and at rest: 100. 38 ± 12. 71，P ＜ 0. 01; in eliminated dog group，heart rate at test: 124. 83 ± 15. 57 and at rest: 98. 41 ± 12. 38，P ＜ 0. 01) ． The difference between heart rate in test and in rest ( △HＲ) was also analyzed between guide dogs and eliminated dogs． The △HＲ of eliminated dogs was significantly higher than that of guide dogs ( eliminated dog group: 26. 42 ± 3. 68，guide dog group: 16. 38 ± 2. 03，P ＜ 0. 05) ． Conclusion The △HＲ of guide dogs was lower compared with that of eliminated dogs． This result suggests that guide dogs are more easily to control fear than eliminated dogs． This study provided an effective reference for early screening of potential guide dogs．

Determination of Blood Physiological and Biochemical Parameters and Organ Indices of Tiantanbio: NIH Mice

2016, 33(06): 45-49.

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Abstract: Objective Blood physiological and biochemical parameters and organ indices of Tiantanbio: NIH mice were determined and analyzed by difference and correlation analysis methods． Method 4 － week and 8 － week old SPF Tiantanbio: NIH mice ( n = 30 each age) ，half male and half female，were chosen as objectives． After anesthesia，blood was collected for determination of 17 physiological and 13 biochemical parameters． Then the mice were dissected to determine organ weights and coefficients． Differences between groups and correlations of organ weights with body weights were analyzed respectively．Ｒesult Ｒesults of difference analysis showed that some blood physiological parameters，most biochemical parameters，organ weights and coefficients were affected by both sex and age． Among physiological parameters，WBC，leukocyte subsets，and blood platelet related indices were included． All the organ weights showed significantly positive correlations with body weights excluding thymus，and female liver weights and male kidney weights had close correlation with body weights． Conclusion Both sex and ages could affect blood physiological and biochemical parameters and organ indices． And there were significant differences between mice of different origins，which asked the producers to establish their own biological characteristic data bases．

Evaluation of a Case about Using IWT650 Cage Washer in a Laboratory Animal Facility

2016, 33(06): 50-51.

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Abstract: In order to improve work efficiency，reduce workload and make sure cleaning quality of cage，we take advantage of the reliability，simplicity and high efficient of IWT 650Cage Washer in daily cage-washing job and evaluate the application． The result dedicates that the desired objectives are achieved and IWT 650 Cage Washer can satisfy the need of particular laboratory animal facilities．

Laboratory Animal Quality Control and its Ｒoles in Teaching andＲesearch in Medical Colleges and Universities

2016, 33(06): 52-54.

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Abstract: Laboratory animals are the necessary supporting conditions for teaching and research in medical colleges and universities，and good quality control of laboratory animals is an important guarantee for teaching and research work． Currently the level of laboratory animal quality control is vary among medical colleges and universities and universities related to biomedical teaching and research in China，which reflects the comprehensive strength of their quality of teaching and research． In this paper we will mainly introduce the core contents of the laboratory animal quality control and hope to provide references for the teacher and researchers in medical colleges and universities．

Studies on the Dynamic Changes on Indicators of Environment of the Barrier Facilities

2016, 33(06): 55-58.

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Abstract: Objective To study dynamic environment status of laboratory animal facilities． Method The environment indicators were detected dynamically in the barrier facilities according to the national standards of laboratory animal．Ｒesult Some environmental indicators are changed， some are unchanged compared to no animal status． Conclusion Dynamic environmental indicators monitoring provides important data for the maintaining of the barrier facilities．

Ｒodent and Non-human Primate Models of Alzheimer＇s Disease Ｒelated Gene Mutation

2016, 33(06): 59-63.

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Abstract: There are 8 － 10 million Alzheimer’s disease ( AD) patients in China at present． The number of patients is growing with population ageing year by year，however，there are still no effective ways to cure and reverse AD progression currently． The effective drugs screened for curing AD based on rodent animal models show poor effects or serious side effects on human trials，this mainly because the pathological and behavioral characteristics are significant different between the rodents animal models and AD patients． The brain and nervous system of nonhuman primate are highly similar to human，thus establish nonhuman primate model of AD is necessary and important． The AD etiology is complex，but the pathogenic gene mutations are clear documented． Hence it is time to pay attention on establishing nonhuman primate animal model of AD using the gene modification techniques or genetic screening analysis． This review mainly described the characteristics and status of AD pathogenic gene related genetically modified mice models and non-human primate animal models of AD．

Development and Prospect of Laboratory Animals Toys in China and Abroad

2016, 33(06): 64-66.

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Abstract: As an essential condition of scientific experiments，laboratory animals have performed a conducive role in the development of life sciences since its emergence． Owing to the increasing levels of concept of protecting animals’rights in the whole world ，the protection of animal welfare has become a highly common issue． In this context，animals’toys，as a sort of animal welfare，appear to be given a significant attention． This essay will evaluate the current situation and prospect of laboratory animals’toys．

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