Table of Content

31 October 2017, Volume 34 Issue 05

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Analysis of Mylpf Expression in 6 Tissues from Spontaneous Diabetic Mongolian Gerbils

2017, 34(05):  1-5. 

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Abstract: Objective To analysis of Mylpf expression levels in six tissues of diabetic Mongolian gerbils and to explore the molecular mechanism of diabetes in Mongolian gerbils． Method We selected a diabetic group and a control group，each group has 6 Mongolian gerbils． Mylpf mＲNA and protein expression level in skeletal muscle， liver，adipose，kidney，heart and brain were separately detected using Ｒeal-time PCＲ and Western blotting．Ｒesult The Ｒeal-time PCＲ showed that compared with control animal，mＲNA expression of Mylpfin skeletal muscle，heart and brain was increased in diabetic group． The relative expression level in diabetic group exhibited decreased compared with control in adipose and liver． And there were no differences between two groups in the kidney． The Western blotting result display that protein of Mylpf in skeletal muscle was significantly increased in diabetes group． The expression of Mylpf in diabetic Mongolian gerbils heart and adipose was lower than control group． Mylpf protein was almost undetectable in liver and brain tissue． And Varied expression was observed in kidney，but there is no difference． Conclusion There is a correlation between Mylpf and the occurrence of diabetes in Mongolian gerbils and theresult show that the effect of Mylpf mainly occurred in skeletal muscle and heart tissue of diabetic gerbils．

Establishment and Application in Bovine Origin Samples of PCＲ Method for Detection of Bovine Herpesvirus Type Ⅰ

2017, 34(05):  6-12. 

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Abstract: Objective To establish a PCＲ method for detection of Bovine herpesvirus type Ⅰ( BHV-1) in cow． Method The primers were designed and synthesized according to the published BHV-1 specific sequences of gB gene． PCＲ method was established，and the specificity，sensitivity，stability of this method were tested． Then the method was used to detect 64 cow’s clibical samples． Ｒesult The developed PCＲ method was no cross reaction with Bovine parainfluenza virus type Ⅲ ( BPIV3) ，Porcine pseudorabies virus ( PＲV) ，herpes simplex virus 1 ( HSV-1 ) ， Canine herpesvirus ( CHV ) ; its minimum detection limit the lowest detection virus titer is 5lg TCID50 /mL，the corresponding template DNA concentration was 5. 26 × 102 copies; BHV-1 DNA maintained at － 30 ℃ for 12 months，still can enlarge the fragment of about 247 bp visible purpose belt． The 145 cow’s clibical samples were 22 positive，and 123 negative about BHV-1 detected by PCＲ． Conclusion The developed PCＲ method is good in specificity，sensitivity，stability，and can be used for detecting the BHV-1 in cow’s clibical samples．

Establishment of TaqMan Ｒeal-time ＲT-PCＲ Method to Detect Mice Encephalomyocarditis virus

2017, 34(05):  13-17. 

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Abstract: Objective To develop a real-time ＲT-PCＲ assay ( rＲT-PCＲ) for efficient detection of Murine Encephalomyocarditis virus ( EMV) ． Method According to the genomic sequences of EMV strain PEC9 ( DQ288856. 1) download from NCBI and to find the conserved sequences． One pair of the specific primers and one TaqMan probe were designed． Then reaction parameters were optimized to develop a real-time ＲT-PCＲ assay ( rＲTPCＲ) ． Ｒesult The method showed a good linear relationship of standard curve with a r 2 value of 0. 99． The sensitivity of the real-time PCＲ was less than 1 copies /μL． 1 000 times higher than ordinary PCＲ method ，and its specificity is strong，common mice strains had no specific amplification，between batch and batch of variation coefficient is less than 2% ． 120 mice feces samples epidemic materials provided by the Shanghai institute of experimental animals were detected by the real-time PCＲ． All the 120 mice samples were negative． Conclusion The real-time ＲT-PCＲmethod is good in specificity and sensitivity，which can make a powerful technical support for EMV epidemiological investigation and detection．

The Curcumol Inhibitory Effect on HNE1 cell of Nasopharyngeal Carcinoma in Tumor-bearing Mouse Mode

2017, 34(05):  18-22. 

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Abstract: Objective To study effects of Curcumol inhibitory on HNE1 cell of Nasopharyngeal Carcinoma( NPC) in Tumor-bearing Mouse Mode． Method The effect of different concentrations of Curcumol ( 12. 5 μg /mL， 25 μg /mL，50 μg /mL，100 μg /mL) on migration ability of HNE1 NPC cells by cell scratch test in vitro and in vivo，HNE1 cell of NPC being vaccinate on nude mice got nasopharyngeal carcinoma． Establish Tumor-bearing Mouse Mode by transplantation with Nasopharyngeal carcinoma on subcutaneous． Curcumol gavage nude mice dose is 0. 79 mg /10g in experience group，control group by gavage water，positive group by gavage doxifluridine． To measure the body and tumour weight twice one week，the mice were killed to collect the samples of tumor on the 30th day． Weigh and Calculation volume( V = a·b·c·π/6) for the rate of inhibitory． Evaluate inhibitory effect of the Curcumol inhibitory on HNE1 cell of NPC in Tumor-bearing Mouse Mode． Ｒesult In vitro，100 μg /mL Curcumol has obvious inhibitory effect，which compared with the control group，has statistically significant( P ＜ 0. 05) ． In vivo，Curcumol has obvious inhibitory effect comparing with the control group，the inhibition rate is 34. 04% ，compared with the control group，the mass of tumor has statistically significant( P ＜ 0. 05) ． The liver and spleen coefficient of the tumor-nude mice of the Curcumol group，which compared with the water group and doxifluridine group has statistically significant( P ＜ 0. 05) ． The spleen coefficient of the tumor-nude mice of the Curcumol group，which compared with the doxifluridine group has statistically significant( P ＜ 0. 05) ． Conclusion The Curcumol has the function of inhibitory on Nasopharyngeal Carcinoma and which provides the experimental basis for the Curcumol developing in anti-anticancer drugs．

The Study of Curcuma Aromatica Volatile Oil on The Learning and Memory Function in Mice Induced by β-amyloid( Aβ25 － 35 )

2017, 34(05):  23-27. 

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Abstract: Objective To investigate the effects of curcuma aromatica volatile oil on learning and memory in mice induced by β-amyloid( ( Aβ25 － 35 ) ． Method Sixty mice were divided into six groups randomly: sham group，model group，curcuma aromatica extract low dose group ( 0. 016 mL / kg) ，high dose group ( 0. 048 mL / kg) ，donepezil group ( 1. 30 mg / kg) and shenzhiling group ( 2. 6 mL / kg) ． β-Amyloid protein ( Aβ25 － 35 ) was infused into the hippocampal of mice． Drugs were administrated for 15 days． Y maze and Morris water maze was detected to evaluate the cognitive behavior． HE-staining was used to observe the neuron changes caused by curcuma aromatica volatile oil． Ｒesult Compared with the model group，curcuma aromatica volatile oil increase the alternation behavior significantly，decrease swimming escape latency and total distance，prolong the swimming distance percentage( P ＜ 0. 05 or P ＜ 0. 01) ; It also can improve the disorder，edema and other symptoms of hippocampus neurons，reduce the death of neurons． Conclusion Curcuma aromatica volatile oil can improve the effects of learning and memory in mice induced by β-amyloid( Aβ25 － 35 ) ．

Rapidly Detecting Avian Leucosis Virus through Establishing Luminex Liquid Chip Technology

2017, 34(05):  28-30. 

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Abstract: Objective To establish the liquid phase chipmethod of rapidly detecting avian leucosis virus，and provide technical basis for the detection of large quantities of samples． Method Prepare matching monoclonal antibody against P27 protein of avian leucosis virus，biotin labelling antibody，magnetic beads coupling capture antibody． this method is validated with Luminex200 analysis system to test specificity，sensitivity and repeatability．Ｒesult The positive criterion is more than twice as much as the blank control． The specificity test result showed that there was no cross-reaction between many other pathogens of the chicken，meaning good specificity． Simultaneously repetitive experiment shows the coefficient of variation is within 7% ，which indicates that the stability of the experiment is good and repeatable． Conclusion The Luminex liquid chip technology，which directly detects the ALV pathogen，is specific，stable and repeatable．

Transcriptional Activity Analysis of Beagle EＲβ and Splice Isoforms EＲβ419，EＲβ43

2017, 34(05):  31-36. 

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Abstract: Objective To detect the transcriptional activity of EＲβ，EＲβ419 and EＲ431 expressed vector． Method The cells were divided into 6 groups，control groups，LV5-EＲβ groups，LV5-NC groups，Lenti-OE-Flag-EＲβ431 groups，Lenti-OE-Flag-EＲβ419 groups，Lenti-OE-Flag-NC groups． Each group was further divided into two groups， each group of cells was transiently transfected with the recombinant plasmid EＲE-LUC and ＲLUC． In each one group with a final concentration of 10 μmol /L E2，while the other group with an equal volume of anhydrous ethanol． The luciferase activity of each group was tested after adding the stimulation． Ｒesult EＲE-LUC and ＲLUC recombinant plasmid was constructed succesfully． Compared with anhydrous ethanol group，The transcriptional activity of Beagle EＲβ with estrogen treatment was significantly increased( P ＜ 0. 01) ，while there was no significant change neither EＲβ419 nor EＲβ431 ( P ＞ 0. 05 ) ． Conclusion Succesfully constructed transcriptional activation system of Beagle EＲβ，but the partly lack of LBD lead to EＲβ419 and EＲβ431 can’t combine with E2，then transcriptional activity could not be activated．

Experimental Study on Traumatic Brain Injury Model in Ｒats and Evaluate Bcl-2 and Bax in Brain Tissue

2017, 34(05):  37-40. 

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Abstract: Objective To study on preparation of TBI rats model by eCCI，and provide a model preparation method for the TBI experimental study． Method SD rats，which were randomly divided into blank control group，model group 1，model group 2，model group 3 and sham-operation group，were produced as the animal model of brain injury in different degree by eCCI from selected different fighting parameters． The speed was 3 m / s and the depth was 3 mm in model group 1． The speed was 4 m / s and the depth was 3 mm in model group 2． The speed was 5 m / s and the depth was 3 mm in model group 3． Evaluated the model quality through the following: To detect the contents of Bax and Bcl-2 of brain tissue by immunohistochemical method ． Ｒesult The Bax、Bcl-2 in the brain tissue of cerebral cortex of neurons showed weak positive reaction in control group and sham operation group． The Bax、Bcl-2 of damaged brain in model group showed positive reaction and the expression was markedly increased with the increase of hit ( P ＜ 0. 01) ． Conclusion By eCCI TBI rat model can be prepared with regulation of hit speed and depth． The damage of model was related to the hit speed． The method of TBI model preparation has the advantages of easy operation，good control ability，repeatability，low mortality rate，and can according to the need be used to make various animal model for the study．

Ｒapid Detection of Staphylococcus aureus in Laboratory Animals by Loop-mediated Isothermal Technique

2017, 34(05):  41-47. 

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Abstract: Objective To establish a sensitive and efficient method for detecting Staphylococcus aureus in laboratory animals． Method A set of four primers targeting the heat-stable nuclease ( nuc) gene( V01281. 1) of S． aureus were designed and the specifity of the Lamp assay was evaluated by common pathogenic bacteria in laboratory animals． The reaction conditions of LAMP were optimized including Mg2 + concentration、dNTPs concentration and time． Using LAMP and PCＲ method to detect S． aureus in pure culture and artificially contaminated feces and compared their sensitivity． Ｒesult No cross reaction was found in non-S． aureus bacteria and the optimal reaction condition of LAMP were 1. 6 μmol /L of each inner primer，0. 2 μmol /L of each outer primer，2. 0 mmol /L of each dNTP，8. 0 mmol /L Mg2 + and incubated at 60 ℃ for 50 min． In pure culture，the detection limit of LAMP was 9CFU /test( 9 × 103 CFU /mL) compared with 9 × 101 CFU /test( 9 × 104 CFU /mL) of PCＲ． In artificially contaminated feces，the detection limit of LAMP was 4. 49 × 104 CFU /0. 1 g feces compared with 4. 49 × 105 CFU /0. 1 g feces of PCＲ． Conclusion The developed LAMP method is simple、sensitive and specific，and is available for rapid detection of S． aureus in laboratory animals．

Background Data of Blood Pressure and Electrocardiograms on SHＲ and Wistar Ｒats at Different Weeks of Age

2017, 34(05):  48-52. 

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Abstract: Objective To set up background data reference values of blood pressure and electrocardiogram( ECG) in SHＲ、Wistar rats at different weeks of age． Method The values of body weight、blood pressure and ECG were obtained from 12 SHＲ and 12 Wistar rats since they were at 6 weeks of age to 22 weeks of age． Ｒesult The body weights of SHＲ had been lower than Wistar since 6 weeks of age; heart rate，systolic blood pressure，diastolic blood pressure and mean pressure of SHＲ were all higher than those indexes of Wistar rats，blood pressure became steady since 10 weeks; ECG indexes had been stable with the exception of QT interval and ST interval which were a little lower from 6 to 9 weeks; among these indexes，ＲＲ interval、QT interval、ST interval of SHＲ were higher compared with those of Wistar． Conclusion Background data reference of SHＲ and Wistar from 6 weeks to 22 weeks had been initially established，concluding body weight，blood pressure and ECGs．

Nutrition Standardization for Ｒare Minnow ( Gobiocypris rarus) : Artificial Diets and its Suitable Ｒearing Strategies

2017, 34(05):  53-62. 

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Abstract: Objective To explore applicable artificial diets and its suitable rearing strategies for this laboratory fish ( Gobiocypris rarus) and improve its nutrition standardization． Method Artificial diets were designed based on nutrition requirement and tested in growth and reproduction experiments，and effects of different rearing strategies such as temperature，feeding frequency and stocking density on the growth and reproduction were revealed fed with suitable artificial diets． Ｒesult Diets with 25% ～ 40% crude protein could meet the requirement for fast growth of rare minnow，especially the diet with 30% crude protein，6% crude lipid and 12. 5 kJ/g gross energy，could also promote the reproductive performance，and the success rate of reproduction could achieve 69. 4% compared to those fed with red worms ( Chironomus larva) ． The spawning interval decreased with increasing dietary protein． The growth rate increased with increasing water temperature in 24 ～ 28℃，but descended with over high temperature． The optimal culture temperature was 24℃ when feeding 1 ～ 3 meals a day． The growth rate decreased with increased stocking density，the feed efficiency was higher in 0. 8 fish /L and the optimal stocking density was 1. 6 fish /L． Conclusion The artificial diets with 30% crude protein，6% crude lipid and 12. 5 kJ/g gross energy could meet the requirement for fast growth of rare minnow，also，the parent fish could reproduce regularly． Too high or too low temperature could affect the growth of rare minnow adversely． High feeding frequency could not always promote the growth and feed conversion rate． The optimal culture temperature for rare minnow was 24℃ to 26℃，and feeding frequency was 2 meals per day． Stocking density obviously affected the growth and gonad development of rare minnow，the optimal density in recirculating aquaculture system should be controlled at 1. 6 fish /L or lower．

A Teaching Practice on Laboratory Animal Ethnics in Medical Laboratory Science

2017, 34(05):  63-69. 

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Abstract: Objective To provide a teaching experience to animal ethnics in medical laboratory science． Method The experience in teaching toxicology practice of 12 years including the contents，and method were summarized and analyzed． Ｒesult The reasonable teaching system of animal ethnics of medical laboratory science was established． Several teaching method were used． Good effects on study were achieved． Conclusion These experiences are valuable references for teaching animal ethnics in medical laboratory science．

A Teaching Practice on Laboratory Animal Ethnics in Medical Laboratory Science

2017, 34(05):  67-69. 

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Abstract: Objective To provide a teaching experience to animal ethnics in medical laboratory science． Method The experience in teaching toxicology practice of 12 years including the contents，and method were summarized and analyzed． Ｒesult The reasonable teaching system of animal ethnics of medical laboratory science was established． Several teaching method were used． Good effects on study were achieved． Conclusion These experiences are valuable references for teaching animal ethnics in medical laboratory science．

Selection of Experimental Animal of Modeling Animals for Gout

2017, 34(05):  70-72. 

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Abstract: : Gout animal models in biomedical research are experimental animal objects and material which are established to simulate human diabetic features． By studying of gout animal models，it’s helpful to recognize the occurrence and developing rule of human diabetes，and then to the forming of prevention and cure method ． In this paper，the research development of various types of gout animal models and the choice of modeling animals were presented and discussed systematically．