Table of Content

25 February 2017, Volume 37 Issue 1

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Reduction of Cardiac Function Induced by FGF21siRNA in Type 1 Diabetic Mice

CHEN Cui, MENG Zhe-ying, ZHENG Yuan-yi, Hu Bing, SHEN E

2017, 37(1):  1-5.  DOI: 10.3969/j.issn.1674-5817.2017.01.001

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Objective To investigate the effects of Fibroblast growth factor 21 (FGF21) siRNA on cardiac function in type 1 diabetic (T1DM) mice and explore the mechanisms of actions. Methods Sixteen C57BL/6J mice were randomly divided into, control group and FGF21siRNA control group, sixteen T1DM model mice were divided into T1DM group and T1DM + FGF21siRNA group, type 1 diabetes was induced by STZ. Cardiac function was analyzed by echocardiography, morphological changes of cardiomyocytes were observed by HE staining, collagen fiber was observed by Masson staining, the expression of α-myocin heavy chain (α-MHC), β-myocin heavy chain (β-MHC), atrial natriuretic peptide (ANP), type I collagen (Col I) and type III collagen (Col III) of myocardium were detected by Real-time PCR. Results The expression of α-MHC, β-MHC, ANP, Col I and Col III in myocardium of T1DM mice treated with FGF21siRNA were increased compared to those of untreated ones. Cardiac function, cardiomyocyte hypertrophy and collagen fiber proliferation of treated mice were worse than those of untreated ones. Conclusion FGF21siRNA may contribute to cardiac malfunction T1DM mice, which may associate with the development of myocardial hypertrophy, myocardial fibrosis.



Expression of ND3 in 5 Tissues of Hereditary Diabetic Mongolian Gerbils

LI Yin-yin, GONG Jing-jing, WU Shao-liang, LI Xiao-hong, WANG Cun-long, HUO Xue-yun, LU Jing, LV Jian-yi, LIU Xin, GUO Meng, LI Chang-long, CHEN Zhen-wen, DU Xiao-yan

2017, 37(1):  6-10.  DOI: 10.3969/j.issn.1674-5817.2017.01.002

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Objective To investigate the mRNA and protein expression level of NADH dehydrogenase 3(ND3) gene in 5 different tissues of diabetic gerbils, to understand the location and target tissue of ND3 in diabetic gerbil model. Methods Six gerbils for control and diabetic groups were selected respectively.The kidney, liver, skeletal muscle, brain and heart from each group were collected. The mRNA and protein expression of ND3 were evaluated by Real time-PCR and Western blotting in both groups. Results The mRNA expression of ND3 in diabetic group was extremely significantly increased in kidney, and significantly decreased in liver and skeletal muscle compared with those of control animal. While it just showed decreasing tendency in brain and increasing tendency in heart tissue. The detection of protein expression showed that the protein level was consistent with mRNA expression in all tissues except heart. Conclusions The kidney may be the target tissue of ND3 in diabetic gerbil. In addition, ND3 also exhibits effect in other 4 tissues of diabetic gerbil.



Pathological Change of Kidney in Pkd1 Knock-out Mice with Polycystic Kidney Induced by Tamoxifen

ZHOU Wei-min, ZHU Ke-yan, CHEN Fang-ming, CAI Yue-qing, FANG Ming-sun, Qi Yue-han, CHEN min-li

2017, 37(1):  11-14.  DOI: 10.3969/j.issn.1674-5817.2017.01.003

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Objective To explore the change of kidney function of polycystic kidney disease (PKD) mice before and after the knockout Pkd1f/f: Cre gene induced by Tamoxifen induction. Methods The Pkd1f/f:Cre transgenic mice, Pkd1f/f:Cre wild type mice aged on 11 days were induced by Tamoxifen,the contents of serum creatinine and urea nitrogen were detected at 7 days, 14 days and 21days after induction. The body weight, kidney weight and the kidney viscera coefficient were detected after 21 day. Result After 7 days induction, the contents of serum creatinine and urea nitrogen began to rise after 14 days and 21 days induction, the parameters were significantly increased (P<0.01) After 21 days induction,the kidney coefficient of knocked out transgenic mice kidney coefficient was higher than that of wild type mice. Conclusion The renal function of Pkd1f/f:Cre transgenic mice were obviously changed by Tamoxifen induction, the kidney of transgenic mice showed symptoms of a large number of vesicles.



Effect of Lateral Ventricle Injection of Ganglioside Sodium on Learning and Memory in Rats with Cerebral Palsy

YANG Feng-xiang, ZHANG Xiao-qiao, LUO Xiu-ling

2017, 37(1):  15-19.  DOI: 10.3969/j.issn.1674-5817.2017.01.004

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Objective To study the mechanism of Ganglioside (GM) lateral ventricle injection on learning and memory ability of rats with cerebral palsy, and to provide theoretical basis for clinical treatment of cerebral palsy. Methods Thirty-six SPF rats were randomly divided into blank control group, model group and GM group. The model group and GM group were randomly divided into the model group and the SD group. After modeling, 1 days and 10, 20, 30 days with the brain localization of rat instrument by rat intracerebroventricular injection of GM administered after modeling and administration after the water maze experiment instrument to explore, navigation, suspension and slope experiments comparing the ability of learning and memory of rats and exercise the ability to change space. And using the ELISA method for the determination of acetylcholine in hippocampus (Ach) and orbital venous blood acetylcholinesterase (AChE) activity. Determination of brain tissue and calmodulin dependent protein kinase II (CaMKII) and calmodulin (CaM). Results Compared with the model group, the rats in group GM after treatment, the time of finding the platform and slope experiment time was significantly shortened, suspension time, the first quadrant and the residence time in the voyage prolonged; positioning latency, swimming distance shortened, the Ach level was decreased, and AChE activity was increased. the brain tissue of liquid CaMK and CaM increased (P<0.05). Conclusion In rat cerebral palsy model, GM could inhibit the synthesis of CaMKII and CaM, reduce hydrolysis of neuronal calcium protein and calcium influx. By blocking the signaling pathway of CaMKII, GM treatment increases the activity of Ach, reduces brain damage caused by cerebral ischemia and hypoxia, thus promoting repair and regeneration of lesioned nerves and subsequently learning and memory.

Establishment of Nonalcoholic Fatty Liver Fibrosis Model and Expression of Inflammatory Factors in Mice

YANG Hua, ZHAO Ya-Juan, OU Qiang

2017, 37(1):  20-24.  DOI: 10.3969/j.issn.1674-5817.2017.01.005

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Objective To establish the non-alcoholic fatty liver fibrosis model in mice and observe the pathological changes and the expression of inflammatory factors. Methods Twenty male C57BL/6J mice were randomly divided into control group and model group. The mice in control group were fed with methionine and choline-supplement(MCS) diet, and the model group were fed with methionine choline deficiency (MCD) diet. The mice were weighed and sacrificed at after 8 weeks feeding. The histopathological changes in liver were observed. The level of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), cholesterol (TC), triglyceride (TG), tumor necrosis factor -〈 (TNF-〈) and interleukin (IL-6 ) were detected. Results The body weight of mice in the model group decreased significantly at the end of the study. Severe hepatic steatosis, liver cell edema, visible focal necrosis and focal lymphocytic infiltration, interface hepatitis, periportal enlargement, hyperplasia of fibrous tissue were observed.The scores of liver inflammation activity and fibrosis score in model group were significantly higher than those in control group (P<0.01).The level of serum ALT、AST、TNF-〈、IL-6 were significantly higher in the model group than that of control group (P<0.01). Conclusion After feeding mice with MCD diet feeding for 8 weeks, non-alcoholic fatty liver fibrosis model of mice was successfully induced. The modeling method was simple, rapid, and with has high survival rate. It is suitable for the pathogenesis and drug intervention study of nonalcoholic fatty liver fibrosis.



The Single Nucleotide Polymorphism Genotyping of Inbred Mice from Shanghai

HAN Lin, XIE Jian-yun, YANG Fei, HU Ying, TIAN Li-li, BAO Shi-min

2017, 37(1):  25-31.  DOI: 10.3969/j.issn.1674-5817.2017.01.006

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Objective To analyze the single nucleotide polymorphism (SNP) genotyping of inbred mice from Shanghai. Methods 44 SNP loci on 21 chromosomes of 10 inbred mice strains from 2 laboratory animal companies of Shanghai were detected by the PCR-LDR genotyping panels, and the other 2 inbred mice strains were selected as the double blind samples for verification. And discrepancy locus were verified by sequencing. Results The SNP genotyping results of all the samples from the same inbred mice strains were homomorphism, but 6 inbred mice strains from different populations(BALB/c, FVB, DBA/2 and C3H/He) were polymorphism with 7 and 2 discrepancy locus respectively. The sequencing results to verify the discrepancy locus were the same as the SNP genotyping results. Conclusions The SNP genotyping was very stable and consistent in inbred mice strains from the same population in Shanghai, and the SNP genotyping of the same inbred mice strains from different populations was polymorphism.

Development of Dual PCR for Detection of Rat Parvovirus

RAO Dan, ZHU Yu-jun, WU Miao-li, YUAN Wen, WANG Jing, YIN Xue-qin, CONG Feng, LIAN Yue-xiao, HUANG Bi-hong, XU Feng-jiao, LIU Xiang-nan, LIU Zhu-hong, HUANG Ren, ZHANG Yu, GUO Peng-ju

2017, 37(1):  32-35.  DOI: 10.3969/j.issn.1674-5817.2017.01.007

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Objective To establish a PCR method for rapidly detecting rat parvovirus. Methods According to the characters of nucleotide sequence of rat parvovirus, a dual PCR was developed to differentially detect RPV and other three rat parvovirus H-1/KRV/RMV. A pair of primers amplifying VP2 gene were applied to exclusively amplify RPV while another pair of primers targeting the NS1 gene was designed for specific amplification of H-1, KRV and RMV primer. Result Rat parvovirus were screened without detecting minute virus of mice which had high nucleotide homology with rat parvovirus and also negative for three other microorganism pathogen. Sensitivity tests showed that the minimum detectable concentration was as low as 1 000 copies μL. When dual PCR combined with sequencing were applied to detect 23 clinical samples, seven samples detected were RMV positive including one coinfected with RPV. Conclusion The dual PCR was verified to be specific and sensitive which could be used as a reliable method to screen rat parvovirus.



Effect of Different Factors on Cyclophosphamide Induced Immunosuppression Mice Model

SONG Ying, GUO Ya-juan, HUANG Ming-qian, LIANG Jin-qiang, HUANG Zhi-ying

2017, 37(1):  36-39.  DOI: 10.3969/j.issn.1674-5817.2017.01.008

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Objective To investigate the different dosages of cyclophosphamide (CY) and endpoint-detection on the effects of immunosuppression model. Methods BALB/c mice were divided into 4 groups randomly: negtive control group (A), CY 40 mg/kg group (B), CY 60 mg/kg group (C), which were all tested at 5 days after the last administration, and CY 60 mg/kg group-2 (D) was tested at 3 days after. The animal’s body weight was measured every day, hematological parameters and immune organ index were detected when the experiments finished, as well as immune function was mediated by T or B lymphocytes testing. Results The different doses of CY and endpoint-detection have impact on the immunosurpression effect. 60 mg/kg CY has stronger immunosurpression effect when tested at 5 days after the last administration. The alteration of immune indicators were more obervious when tested at 3 days after the last administration compared with 5 days at the dosage of 60 mg/kg CY. Conclusion Dosages of CY and endpoint-detection have impact on immunosuppressed mice model. 60 mg/kg CY is more effective than 40 mg/kg CY, and the results are better when 3 days after the last administration is chosen as the endpoint-detection.

Improvement on Tail Tip Bleeding Model in Mice

TIAN Jing, CUI Qing-hua, GUO Ping, ZHANG Zhi-xiao, FENG Zi-liang, ZHENG Ying, FAN Quan-shui

2017, 37(1):  40-45.  DOI: 10.3969/j.issn.1674-5817.2017.01.009

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Objective The application of tail tip bleeding model in mice was explored to evaluate the hemostatic activity of some coagulation components. Method This tail tip bleeding model improved to be used as a trauma model in the aspect of coagulant evaluation was compared with the traditional operation process. The new method was established for fast quantitative detection of the blood loss and decided the bleeding time from the starting point when the tail was cut off to the end point when the blood loss was less than 1 μL in two minutes intervals. Saculin approved as a coagulant drug on sale in domestic market was used to verify the effectiveness of this assay. The sensitivity between coagulate time and blood loss (or bleeding time) is also compared in mice. Results The advanced tail tip bleeding model in mice can quantitative analysis blood loss and bleeding time in order to reduce the personal error to a great extent. This model is easy to use and can simulate the process of trauma properly. Conclusion The tail tip bleeding model in mice can reflect the action of the coagulant drug sensitively, which is a useful tool for screening and assessment of coagulants.



Effects of Single Caged and Social Feeding on Emotion of Female Macaca Fascicularis

LI Wei, JIN Yi, XING Zheng-hong, CHEN Guo-qiang

2017, 37(1):  46-49.  DOI: 10.3969/j.issn.1674-5817.2017.01.010

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Objective To understand the differences on serum monoamine neurotransmitters (5-hydroxytryptamine, norepinephrine, dopamine) in single caged and social group female macaca fascicularis and ECG, blood pressure which were associated with the emotion, explored the influence of living environment on the emotion of macaca fascicularis. Methods Nine female macaca fascicularis at the age of 4-6 years old were singly raised in cage at the first 5 months, and group raised in next 5 months.The ECG, blood pressure, norepinephrine (NA), dopamine (DA), 5-hydroxytryptamine (5-HT) were measured,daily facial expressions also were recorded. Results During the whole experiment, the indexes of ventricular rate, PR interval, QT interval, serum monoamine neurotransmitter (NA, 5-HT and DA) and diastolic pressure; value were compared between groups difference significant (P<0.05 or P<0.01). Anger and anxiety score in single caged stage were significantly higher than in group phase (P<0.05), but sexual behavior, activity and happiness score were lower (P<0.05). Conclusion The macaca fascicularis was more suitable for the breeding of group,during long-term single caged feeding, it could have a greater impact on the serum monoamine neurotransmitters and some indicators of ECG, blood pressure,and were prone to a variety of negative emotions.

Reproductive Performance of Three Breeds of Minipigs

YANG Li-chang, ZHOU Wen-bing, DING Jun, ZHOU Xin-chu, XIE Dong, YANG Wei-min

2017, 37(1):  50-54.  DOI: 10.3969/j.issn.1674-5817.2017.01.011

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Objective To determine the reproductive performance of Bama minipigs, Guizhou minipigs and Banna minipigs reared in Shanghai. Method The data of reproductive performance of Bama minipigs, Guizhou minipigs and Banna minipigs such as puberty age, sexual mature age, breeding age, and so on were combined from farm and laboratory, and then compared the results with one-way ANOVA among three breeds. Result Among ten reproductive parameters of female Bama minipigs, Guizhou minipigs and Banna minipigs, litter weight at birth and survival rate at weaning between Guizhou minipigs and Banna minipigs were significant different (P<0.05). Sexual mature age among the three breeds of minipigs (Bama, Guizhou and Banna minipigs) were significant different (P<0.05); breeding age among the three breeds of minipigs were close, and average body weight at breeding of Bama minipigs was lightest and Bana minipigs was heaviest. Conclusion Determination of the reproductive performance of Bama minipigs, Guizhou minipigs, and Banna minipigs will provide the basis data for genetic breeding of minipigs and background data for laboratory animals industrializing.

Establishment of Urine Collection Method and Determination of Some Biochemical Indexes for Meriones Meridianu in Xinjiang

XU Yi-mei, SHI Shen, LUO Yun, YAN Shun-sheng, ZHAO Hong-qiong, LIAO Li-fu

2017, 37(1):  55-58.  DOI: 10.3969/j.issn.1674-5817.2017.01.012

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Objective To establish the method for urine collection, detect and analyses the urine biochemical and electrolyte index of Meriones meridianus in Xinjiang area, and compare the index with SD rats. Methods Wild Meriones meridianus was reared in the laboratory for three months, to adapt to the laboratory artificial rearing conditions. The urine of selected 40 Meriones meridianus and 40 clean grade SD rats was collected using simple urine collection cage to analyze urine biochemical parameters and electrolyte index. Results The amount of urine of Meriones meridianus and 1.5 months aged SD rat in 24h was 3-3.5 mL and 8-10 mL respectively. The urine of Meriones meridianus and SD rat’s was yellow, transparent, odor of ammonia by visual observation. The pH of Meriones meridianus’s urine was 7.80 ±0.40, urine specific gravity was 1.011 ± 0.005. The pH of SD rats’s urine was 7.12±0.03, urine specific gravity was 1.018±0.003. The results of biochemical test showed that the most of Meriones meridianus and SD rats are negative, but there were individual animal samples with different degrees of positive. All urine electrolytes showed no significant difference between the negative index of Meriones meridianus (P>0.05), the urine electrolytes showed no significant difference between male and female (P>0.05). Conclusions The results provided basic biological information and research on Meriones meridianus as experimental animal.



PI3K/Akt and AMPK Signaling Pathway and Effect of Exercise on Rodent Skeletal Muscle GLUT4 Translocation and Expression

ZHANG Yun-li, WANG Lin, LIU Tie-min

2017, 37(1):  76-82.  DOI: 10.3969/j.issn.1674-5817.2017.01.017

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Skeletal muscles play an important role in glucose homeostasis. The translocation and expression changes of glucose transporter 4 (GLUT4) as one of the most important glucose transporter proteins in skeletal muscle are closely associated with insulin resistance. This paper reviewed the recent research progress on the exercise-induced translocation and expression of GLUT4 as well as the improvement of exercise-induced glucose uptake by phosphatidylinositol 3-kinases (PI3K) / protein kinase B (Akt) and AMP-activated protein kinase (AMPK) signaling pathways in rodent skeletal muscle. The purpose of this paper is to clearly understand the mechanism underlying GLUT4 translocation and expression in rodent skeletal muscles.