Table of Content

25 February 2018, Volume 38 Issue 1

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Analysis of Microsatellite Genetic Characteristics in of Closed Colony of Tree Shrews

LIU Cheng-xiu, LI Na, TONG Pin-fen, WANG Wen-guang, LU Cai-xia, HAN Yuan-yuan, KUANG De-xuan, SUN Xiao-mei, DAI Jie-jie

2018, 38(1):  1-9.  DOI: 10.3969/j.issn.1674-5817.2018.01.001

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Objective To find out the genetic variation and differentiation of the I~IV generations in tree shrews (Tupaia belangeri chinensis),and provide genetic bases for closed colony tree shrews.Methods The forty-nine genomic DNA of blood samples were extracted from I~IV generations tree shrews,the amplification products were detected by capillary electrophoresis and marked with fluorescent tags (FAM) PCR,then the genetic diversity of the tree shrew colony was analyzed comparatively using POPGENE,FSTAT software with tree shrews 18 microsatellite loci.Results The 89 alleles were found in the tree shrew colony,the average effective number of alleles,observed heterozygosity,expected heterozygosity,polymorphism information content,Shannon information index and allele richness were 3.779,0.523,0.613,0.558,1.277 and 2.938 respectively.The mean value of Fst was 0.046 in the colony.The genetic distance and unbiased genetic distance were 0.049~0.159 and 0.022~0.109 respectively.Conclusions The closed clony tree shrew shows high genetic diversity,and there was no significant difference in genetic diversity and differentiation.



Genetic Analysis on Two Closed Colonies of SD Rats Using Microsatellite DNA Markers

HONG Yang, YUAN Ming-ming, ZHOU Jing, ZHANG Lei, DONG Han

2018, 38(1):  10-15.  DOI: 10.3969/j.issn.1674-5817.2018.01.002

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Objective To analyze genetic diversity and relationship of two closed colonies of SD rats from Beijing and Shanghai respectively.Methods Seven microsatellite loci were tested by Hardy-Weinberg equilibrium (Hardy-Weinberg equilibrium,HWE),meanwhile,the values of the allele frequency,gene heterozygosity,polymorphic information content (PIC),the genetic distance and some other indexes were counted.Results The average number of observed alleles are 4.857 1 and 5.142 9 in two colonies respectively.The average observed heterozygosity were 0.647 6 and 0.585 7,the average PIC were 0.644 5,0.667 6 respectively.Three and two loci showed evident deviation from that of the HWE (P<0.05) in SD rat colony from Beijing and Shanghai respectively.The unbiased genetic distance of the two colonies was 0.158 2 (Nei 1978).Conclusion The genetic diversity of SD rat colonies from Beijing and Shanghai are relatively rich.There are medium level of genetic differentiation between the two colonies.Microsatellite marker technique is an effective mean for genetic monitoring for closed clolony of SD rats.

Genetic Mornitoring for Inbred Mice with Single Nucleotide Polymorphisms

LI Yin-yin, CHEN Zhen-wen, LIU Li-jun, LI Chang-long, LIU Xin, LV Jian-yi, GUO Meng, DU Xiao-yan

2018, 38(1):  16-21.  DOI: 10.3969/j.issn.1674-5817.2018.01.003

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Objective To detect the genetic quality of 29 inbred strains mice from 36 different colonies using 95 single nucleotide polymorphism (SNP) loci and analyze the different genotype among different strains.Methods All genotypes of samples from domestic including 36 inbred mice colonies such as C57BL/6J,BALB/c were identified by the methods of matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) using 95 SNPs referenced from domestic and international literature.Then the numbers of loci with different genotypes between each two strains were analyzed and compared.Results Totally 29 colonies (80.56%) of the mouse were successfully genotyped with 95 SNPs while very few strains weren’t,because of sample’s quality focusing on 3 strains with the background of BALB/c.Most loci were homozygous and the rate of monomorphic loci within strains was up to 98.95%.The numbers of loci with different genotypes between each two colonies reached maximal 58 loci and minimal 1 locus which distributed in the same strain from distinct sources and the gene modified lines between their backgrounds.Conclusions The selected SNPs could be applied to mornitor and identify the genetic condition of inbred mice.However,SNP loci panel should be optimized on behalf of each strain’s characteristics.

A Melanoma Zebrafish Model for Real-time Imaging in vivo

ZHOU Bin, SHENG Zhe-jin, FENG Chen-zhuo, LI Li-mei

2018, 38(1):  22-28.  DOI: 10.3969/j.issn.1674-5817.2018.01.004

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Objective To establish a zebrafish model for real-time monotoring the development of melanoma in vivo and screening relative drugs.Methods The expression effect of pMITFa1-RedV12/hRas plasmid,in which the melanoma was labeled by red fluorescent protein (RFP),was verified in mouse B16 melanocytes.After microinjecting pMITFa1-RedV12/hRas into 1- cell stage of fertilized eggs of zebrafish,we monitored the process of melanoma formation with in vivo imaging system in zebrafish.Results pMITFa1-RedV12/hRas was successfully expressed in mouse melanoma B16 cells.Three days after microinjection of pMITFa1-RedV12/hRas vector to 1-cell satge of zebrafish,melanomas labeled by RFP protein was observed under the fluorescence microscope.Eight weeks later,melanomas were observed clearly by eyes.The rate of tumor formation was 100%.When microinjected into pMITFa1-RedV12/hRas vector into fertilized eggs of TG(zlyz:EGFP) transgenic zebrafish,the interaction between the melanoma cells labeled by RFP protein and the macrophages labeled by green fluorescent protein was clearly observed in in vivo imaging system.Conclusion Successfully established a zebrafish model for real-time monitoring melanoma in vivo,which is cheap and easy to operate.



Investigation on Mouse Model of Type 2 Diabetes Mellitus Combined with Alzheimer's Disease

CHEN Zhi-ling, WU Hua, SONG Xin-lei

2018, 38(1):  29-35.  DOI: 10.3969/j.issn.1674-5817.2018.01.005

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Objective To construct a mouse model of type 2 diabetes mellitus (T2D) combined with Alzheimer's disease (AD).Methods The male (APP/PS1^+/+/db/db^+/+) mice and female (APP/PS1^-/-/db/db^+/-) mice were hybridized.The male and female offspring with genotype (APP/PS1^+/-/db/db^+/-) were mated,and produced the offspring with genotype (APP/PS1^+/+/db/db^-/-) as the model group (group M);with genotype (APP/PS1^-/-/db/db^+/+) and (APP/PS1^-/-/db/db^+/-) as the control group (group C).Thirty mice randomly selected from 2 groups were divided into 3 subgroups with the age of 4,14 and 25 weeks old respectively.The metabolic indexes,such as body weight,fasting blood glucose,postprandial blood glucose,blood sugar insulin content and insulin resistance index,of mice in different groups were detected.The learning and memory ability of mice were detected through the water maze test.The anatomical feature of brain tissue (brain weight and cortical and hippocampal area) were detected via cresyl violet staining,and the microglia,astrocytes,and the content and distribution of A protein and senile plaques (SP) in cortex and hippocampus were detected by immunohistochemistry staining.Results Animals in M group,starting from 4 weeks of age,body weight index was significantly increased compared with C group on blood glucose,insulin content and insulin resistance (P<0.01).While the brain weight,search platform latency,the original platform quadrant time ratio were significantly decreased (P<0.01).At 14 weeks of age,area of cerebral cortex and hippocampus decreased significantly (P<0.01),tau protein phosphorylation level was significantly increased (P<0.01).At 25 weeks of age,cortex and hippocampus A beta deposition,and the formation of SP at the same time,microglia and astrocytes increased significantly (P<0.01).Conclusion The established mouse model of T2D combined with AD,which provided a platform for study of the relationship between the two diseases and development of therapeutic drugs.



Establishment and Application of RT-PCR Diagnostic Method for Enterotropic Mouse Hepatitis Virus

AI Dong-xu, SUN Fei, Li Yu, SUN Zhao-Zeng, LI Lian-rui

2018, 38(1):  36-39.  DOI: 10.3969/j.issn.1674-5817.2018.01.006

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Objective To establish a method for RT-PCR detection of mouse hepatitis virus (MHV) and provide a rapid method for monitoring the daily production process of mice.Methods The total RNA form mouse colon tissue was extracted.The first strand cDNA was synthesized by reverse transcription reaction.Then using the cDNA as the template,a DNA fragment of MHV was amplified by PCR with four pairs of specific primers.The PCR reaction conditions (including primers,primer concentration,annealing temperature,template concentration,etc.) were optimized.Results The two pairs of MHV primers were effective and specific.The PCR results of these two pairs of primers were consistent with the results of ELISA.Conclusion The RT-PCR detection method of MHV was specific and relatively simple.It can be used as a rapid method for MHV detection in the production process of mice.



Influence of Electromagnetic Radiation on Rats Liver under Complex Electromagnetic Environment

LV Zhao-hui, ZHANG Wen-luan, GENG De-jun, FAN Yu-lin, SHENG Xiao-juan, LI Xi-ping

2018, 38(1):  40-43.  DOI: 10.3969/j.issn.1674-5817.2018.01.007

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Objective To investigate the influence of electromagnetic radiation on rat’s health under the complex electromagnetic environment.Methods The Wistar rats were divided into control and treatment groups 1,2,3,and 4.Then they were exposed in complex electromagnetic environments for different certain period per day.After 30 days exposing,the livers of all rats were sampled and studied.Results Compared with the control group,liver cells of experimental rats were confirmed to be occurring apoptosis at both the DNA level and protein level.And the degree of apoptosis was increased with the exposure time.Conclusion Complex electromagnetic environment can damage rat health.



Detection of Whole Methylation Level of Liver Genomic DNA in Mongolian Gerbil Based on Indirect ELISA

WANG Zhi-yuan, LIU Yue-huan

2018, 38(1):  44-47.  DOI: 10.3969/j.issn.1674-5817.2018.01.008

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Objective To detect the genomic DNA methylation level in the liver of Mongolian gerbil by indirect ELISA.Methods Six newborn gerbils (newborn group),12 young gerbils (6 in young control group and 6 in young nonalcoholic fatty liver disease (NAFLD) model group,6 gerbils in the 8-month-old (middle and aged group) were employed,all were male.The serum samples (except newborn group) were collected for detection of the level of total cholesterol(TC),triglycerides(TG),high-density lipoprotein (HDL),low-density lipoprotein(LDL).At the same time,two samples of liver were collected for HE staining and detecting the whole methylation level of liver genomic DNA [ie,5-methylcytosine (5-mC) contentby indirect ELISA kit.Results The levels of serum TC and TG in the young model group and the middle and aged group were significantly difference with those in the young control group (P<0.05).Morphological examination revealed that the young model group and middle and aged groups developed obvious hepatic steatosis.There are a large number of piles of mononuclear cells and piles of erythrocytes in neonatal gerbil liver sinus.Overall methylation test results showed that high fat diet induced model young group gerbils>newborn gerbils>middle and aged gerbils>young control group gerbils.Conclusions The degree of fatty degeneration caused by NAFLD and age in Mongolian gerbil correlated with methylation level of genomic DNA.The method of indirect ELISA for detecting methylation level of liver genome DNA was simple,rapid and economic,which would be developed as a method to evaluate epigenetics of gerbils.



Dynamic Changes of Biochemical and Physiological Indices in Ischemia-prone Inbred Gerbil Strain during Breeding

LI Ying, DU Xiao-yan, CUI Xiao-xia, MA Lan-zhi, SHANG Shi-chen, HE Xiao-ya, HUANG Bin, ZHAO Quan, LI Gui-jun, WANG Dong-ping, CHEN Zhen-wen

2018, 38(1):  48-53.  DOI: 10.3969/j.issn.1674-5817.2018.01.009

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Objective To investigate a dynamic curve of blood biochemical and physiological characteristics in the process of inbreeding ischemia-prone gerbil,and to provide basic data for inbred animal breeding.Methods The 17 blood biochemical and 22 physiological indices of blood samples from inbred 11th to 20th generation (F11 to F20) were detectioned by Beckman automatic biochemical analyzer (CX5) and Japanese photoelectric automatic blood cell analyzer (MEK-7222K).Results The results showed that the uricacidcrystal (URIC),lactate dehydrogenase (LD),creatine kinase (CK),alanine aminotransferase (ALT),high density lipoproteins (HDLD),magnesium (MG),red blood cell (RBC),leukocyte (WBC),eosinophil (EOS),neutrophil count (NEUT),monocytes (MON),basophilic granulocyte(BAS),blood platelet (PLT),thrombocytocrit (PCT),platelet distribution width (PDW),hemoglobin(HGB),mean corpuscular hemoglobin (MCH),and red blood cell distribution width (RDW) of F11-20 gerbil were increasing.The lymphocyte (LYM),creatinine (CRE),blood urea nitrogen (BUN),alkaline phosphatase (ALP),glucose (GLU) showed a decreasing trend.The hematocrit (HCT),mean corpuscular volume (MCV),mean corpuscular hemoglobin concentration (MCHC),average platelet volume (MPV) value was stable.The majority of indicators tend to be stable after F17.Conclusion The gerbil inbred strain has a certain influence on the most of the blood physiological and biochemical indices.



Preliminary Study on Growth and Reproductive Performance of Inbred Mongolian Gerbil Strains CMU/1 and CMU/2

LI Ying, DU Xiao-yan, CUI Xiao-xia, MA Lan-zhi, SHANG Shi-chen, HUANG Bin, ZHAO Quan, LI Gui-jun, WANG Dong-ping, CHEN Zhen-wen

2018, 38(1):  54-56.  DOI: 10.3969/j.issn.1674-5817.2018.01.010

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Objective To investigate the growth and reproductive performance of CMU/1 and CMU/2 ischemia-prone inbred gerbils;And to provide the data support for its’application,conservation and breeding.Methods The generations F20~F22 of 2 gerbil lines were selected and bred in accordance with inbreed Mating rule.The litter interval,the number of birth,the number of weaned pups and the growth curve of the 1~5 litters was analyzed.Results The birth weight of CMU/1 and CMU/2 was 2.5 g to 3.4 g.The litter size ranged from at least 1 to the most 9.The survival rate of weaned litter from 65% to 75%.The shortest litter interval was 14 days,whereas the longest 171 days.The weight gain at 2 to 6 week of age was more rapid.Conclusion There is significant difference in litter size between CMU/1 and CMU/2 ischemia-prone inbred gerbil.Most of them display birth intervals in the 21~70 day.Genotype seems have no effect on the reproductive characteristics of ischemia-prone inbred gerbils.



Comparison of Four Blood Coagulation Factors among Gerbil CMU/1,Gerbil CMU/2 and F344 Rat and BALB/c Mouse

LI Ying, DU Xiao-yan, CUI Xiao-xia, MA Lan-zhi, SHANG Shi-chen, ZHAO Zhi-Bing, ZHAO Quan, LI Gui-jun, WANG Dong-ping, CHEN Zhen-wen

2018, 38(1):  57-59.  DOI: 10.3969/j.issn.1674-5817.2018.01.011

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Objective To detect and compare the differences of coagulation factors between inbred gerbil CMU/1 and CMU/2,inbred rat F344,and inbred mouse BALB/c;then to analyze the characteristics of inbred ischemia-prone gerbil.Methods The four channel blood coagulation analyzer was used to determine three kinds of inbred animals’plasma prothrombin time (PT),activated partial thromboplastin time (APTT),fibrinogen (FIB),and thrombin time (TT).The SPSS23.0 software was applied for statistical analysis of data.Results The APTT FIB of CMU/1 and the FIB of CMU/2 of ischemia-prone gerbil displayed high standard deviation.The PT and FIB of CMU/1 and CMU/2 showed significantly different compared with that of F344 rat.The APTT of both ischemia-prone inbred gerbil lines demonstrated significant difference compared with BALB/c mouse and F344 rat.Conclusion The 4 coagulation factors of ischemia-prone inbred gerbil lines are consistent with those of inbred BALB/c mouse;which are significantly different from those of inbred F344 rat.

Progress of Microtus Fortis in Research and Application of Medical Biology

CAI Shu-mei, FU Zhi-qiang, XIE Jian-yun

2018, 38(1):  72-77.  DOI: 10.3969/j.issn.1674-5817.2018.01.016

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Microtus fortis which has the characteristics of natural resistance to Schistosoma japonicum,has been developed as a unique model animal for schistosomiasis resistance in China.It can also be used as a model animal for the study of diabetes,ovarian cancer and nonalcoholic fatty liver.In this paper,the classification,distribution,morphology and biological characteristics of Microtus fortis were reviewed,and the progresses of the study on the animal models of schistosomiasis,diabetes,ovarian cancer and nonalcoholic fat were summarized.

Research Progress in Animal Model of Cataract

JIA Jie, DAI Jie-jie

2018, 38(1):  78-82.  DOI: 10.3969/j.issn.1674-5817.2018.01.017

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This paper summarized the method for establishing animal models of congenital,traumatic,artificially induced and cellular-level cataract that have been studied at domestic and abroad in nearly resent 20 years.The characteristics,evaluation criteria and related mechanisms of each model were also described.The paper also proposesed a method for studying the expression of gene in lens cells that in different state of cataract by established immortalized lens cell lines,and an assumption that setting up tree shrew model for cataract that are similar to human cataract.