1.Expression levels and targets of more than fifty miRNAs in glioma
Taiqi LIU ; Siyu CHEN ; Wende LI ; Ren HUANG
Chinese Journal of Comparative Medicine 2016;26(7):79-84
miRNAs are a class of small endogenous RNAs that degrade target mRNAs or repress their translation process. Several miRNAs in glioma are up?regulated, while some others down?regulated. Some miRNAs promote tumorigenesis; some others, however, play a similar function of tumor suppressor genes. Therefore, studies on the expression profiles of miRNAs in glioma may afford auxiliary basis for early clinical diagnosis and novel srtategies for therapy of glioma. This paper will review on researches about the expression levels of miRNAs and their targets in glioma.
2.Bcl-2 gene-modified neural stem cell transplantation for spinal cord injury in rats
Mei ZHANG ; Yuexin WANG ; Xiaohua HOU ; Jun HONG ; Shengchun YIN ; Yan LI ; Qingyang LIU
Chinese Journal of Comparative Medicine 2016;26(7):35-41
Objective To investigate the bcl?2 gene modification on neurological function recovery in rats with spinal cord injury in neural stem cell transplantation. Methods Cultured rat neural stem cells by Ad?EGFP as vector?mediated side B?cell lymphoma 2 gene ( bcl?2 ) gene transfection of neural stem cells were divided into 3 groups: control group, negative transfection group, bcl?2 transfection group. Use western?blot to detect the expression of bcl?2 protein in neural stem cells before and after transfection. 85 adult female SD rats, successful model 72, were randomly divided into control group, NSCs group, bcl?2?NSCs groups, 24/group, rat acute spinal cord injury model in accordance with a modified Allen’ s method. Assess the motor function by BBB rating and the swash plate test. 7 days after modeling by RT?PCR and Western blot detection of spinal cord injury around HSP27, c?fos gene expression, TUNEL assay apoptosis. Four weeks after model drawn line HE staining and fluorescence microscopy EGFP?labeled NSC survival and distribution of the rats neurophysiological recovery by SEP and MEP. Results bcl?2 gene transfection of rat neural stem cells, bcl?2 transfection group and control group, negative transfection group compared to bcl?2 mRNA and protein levels were expressed ( P < 0. 05 ); lower extremity motor function in rats evaluation of bcl?2?NSCs group than NSCs group, NSCs group than the control group. 72 hours after modeling, bcl?2?NSCs number of apoptotic cells were significantly lower than the control group and NSCs group (P < 0. 05). 7 days after modeling, compared with the control group and NSCs group, bcl?2?NSCs group HSP27 gene and protein expression was significantly higher than that (P < 0. 05), bcl?2?NSCs group c?fos mRNA and protein expression was significantly reduced compared (P < 0. 05). 4 weeks after modeling, HE staining control group showed spinal cord tissue loss and the formation of syringomyelia, no axonal through. NSCs group damage zone few of neuraxis?like structures, syringomyelia smaller, bcl?2?NSCs group showed more nerve axon?like structure, no syringomyelia. EGFP?positive cells labeled:bcl?2?NSCs group the most, NSCs group followed, no control group, and the difference between the groups was statistically significant (P < 0. 05). After the 4th week, SEP and MEP latency period:bcl?2?NSCs group
3.Effect of cyaniding-3-glucoside on glucose and lipid metabolism in the APP swe/PS1ΔE9 mouse model of Alzheimer?s disease
Nan SONG ; Ling ZHANG ; Wei CHEN ; Qian ZHANG ; Yunlin HAN ; Chuan QIN
Chinese Journal of Comparative Medicine 2016;26(7):15-23
Objective To investigate the effect of cyaniding?3?glucoside (Cy3G) on glucose and lipid metabolism in the APPswe/PS1ΔE9 mouse model of Alzheimer’s disease (AD). Methods Seven?month?old APPswe/PS1ΔE9(PAP) mice were randomly divided into model group (PAP), Cy3G treatment group (PAPCy, 5 mg/kg/d) and negative?control group (nPAP). In addition, age?matched and normal wild?type of C57BL/6J mice were selected and divided into vehicle group (WT), Cy3G intervention group (WTCy, 5 mg/kg/d). Each group containing 12 mice, with equal number of male and female mice. After 8?week Cy3G supplementation, microPET/CT was used to measure cerebral glucose metabolism rate of mice in each group. Biochemical methods were used to detect the liver / kidney function as well as indicators associated with lipid metabolism. After weighting brain tissue, the brain coefficient was tested and pathological examination was used to observe tissues changes. Transmission electron microscope was used to observe neuropathological amyloid plaques deposition. Western?blot was used to determine protein levels of AKT and JNK. Results Compared with the WT group, PAP mice had low levels of 18 F?FDG uptake rates, especially in the regions of the frontal lobe and hippocampus accompanied by the decreased brain coefficient and amyloid plaques deposition in hippocampus. And levels of aspartate transaminase ( AST) and lactic dehydrogenase ( LDH) were also increased in PAP mice, but lipid metabolism index was relatively normal. In addition, the expression of JNK was decreased and AKT was increased in mice of PAP. However, in the PAPCy group, 18 F?FDG uptake rates were obviously increased in the regions of the frontal lobe and hippocampus compared with those in the PAP mice. And the reduction of brain atrophy and amyloid plaques deposition, normal lipid metabolism and no obvious liver/kidney toxicity were also observed. Cy3G also could reverse the changes of JNK and AKT protein. Conclusions Cy3G can improve glucose metabolism disorders instead of lipid metabolism, inhibit the senile plaques deposition in hippocampus and regulate insulin resistance and inflammatory reaction associated with JNK/AKT pathway. Thus, Cy3G has a good safety profile and may be used as an ideal alternative to traditional disease?modifying treatments against AD.
4.The genetic reference population “Collaborative Cross” is a powerful resource for genetic discoveries and understanding complex genetic traits
Chinese Journal of Comparative Medicine 2016;26(7):1-10
The Collaborative Cross( CC) is a powerful genetic resource with a wide range of applications in medical research. It is a multiparental genetic reference population, composed of 200 recombinant inbred mouse strains. It was designed to be a resource for rapid mapping of genes that mediate complex traits. The CC has been produced following an intense breeding program that had taken over ten years to complete. It is now becoming available to the research community. While advances in human genetic technologies over the last decade have been substantial, the CC provides the ability to make discoveries not possible with other methods or resources. It can be applied to any subject that can be measured or modelled in the mouse. Here, we discuss a range of applications for which the CC can be used, with examples taken from the early characterization of this powerful resource.
5.Pathologic study and suggestion on evaluation methods of auxiliary protective function on gastric mucosa injury
Yi JIN ; Yuxi GAN ; Yuanping LIU ; Runhua CHEN ; Tianzhu XIE ; Zeyu HUANG
Chinese Journal of Comparative Medicine 2015;(10):38-41
Objective To establish a scientific and practical principle , grading standard and reasonable statistical method for evaluating the protective effect of health food to gastric mucosal injury , based on general pathology and histopathological diagnosis .Methods A methodological study was conducted on rat model of acute gastric mucosa injury induced by alcoholic through comprehensive analysis and comparing shortcomings of the current standard evaluation method, and methods of semi-quantitative analysis and corresponding information statistic processing were based on characteristics of the lesion and principles of pathology .Results Gross pathological evaluation of gastric mucosa lesion was based on the area occupied and proportion in the whole gastric mucosa .Histopathological diagnosis was based on the mucous layer depth of lesion as main determination point and other lesions as reference factors .Grades of lesion were divided into no abnormality (0), mild lesions (1 point), moderate lesions (2 points), and severe lesions (3 points). Ridit statistical method was used for pathological analysis of semi-quantitative results .Conclusion A scientific and feasible evaluation method for the protective effect of health food to gastric mucosal injury was provided from the aspects of gross pathology , histopathological evaluation method , data processing method and result determination .
6.Effect of natural mountain spring on SOD and MDA content in mice aging
Aijun KANG ; Yuwei LI ; Xuewen WANG ; Jianguo HU ; Junfang LIU ; Zhenhui ZHENG
Chinese Journal of Comparative Medicine 2015;(10):34-37
Objective To observe the effect of Natural mountain spring on oxidative damage of aging mice . Methods Thirty male aging mice were randomly divided into experimental group ( drink Natural mountain spring ) and control group ( drink tap water ) according to the level of MDA .The serum in the two groups was taken for T-SOD activity and MDA content analysis after two months .Results The MDA content in experimental group decreased markedly than that in control group .Conclusion Natural mountain spring could alleviate oxidative damage by free radicals to extent .
7.The synergistic effect of estradiol and testosterone on the lipids and coagulation function in mice with hyperlipidemia
Wen DAI ; Yan LI ; Hongyun ZHENG ; Shuwen XU
Chinese Journal of Comparative Medicine 2015;(10):29-33
Objective To investigate the influence of synergistic effect of estradiol and testosterone on the level of lipids and coagulation function in mice with hyperlipidemia .Methods We established a maouse model in hyperlipidmia , giving estradiol (1 μg/d), testosterone (7 μg/d) or estradiol combined testosterone (1 μg/d E2 +7Tμg/d), respectively.After 14 weeks, we collected the blood from the mice , separated the serum to detect the lipids level , separated the plasma to test the coagulation function .Result Compared with controls , after high-fat diet , the serum level of total cholesterol ( TC) , triglyceride ( TG) , high density lipoprotein ( HDL) were significantly increased , PT and APTT were shorter ( P<0.05 ).However, after treating with estradiol combined testosterone , compared with cases, the serum level of C, TG and HDL were lower and PT , APTT were longer ( P <0.05 ), had no significance with controls . Conclusion The synergistic effect of estradiol and testosterone can mediate the lipids , reduce the level of LDL , regulate the coagulation function , reduce the risk of coronary heart disease , and also provide a new strategy for hormone therapy in coronary heart disease .
8.Development and application of RT-PCR for detection of TMEV
Xiaobo LI ; Rui FU ; Ji WANG ; Li WEI ; Shujing WANG ; Bingfei YUE ; Zhengming HE
Chinese Journal of Comparative Medicine 2015;(10):17-20
Objective To develop RT-PCR for detection of TMEV and apply the method .Methods To design specific primers on the basis of GD VII ( GI:62039) genome sequences published in NCBI and establish RT-PCR.To verify the sensitivity and specificity of method after optimizing PCR .We infected 9 BALB/c mice intracerebrally and collected brain, heart, liver, spleen, lung, kidnet, cecal contents and serum samples the 6th day postinfection.The samples were tested by the TMEV RT-PCR.100 mouse cecal contents samples were also detected to apply the established method . Results The 371bp single band was amplified using GDVII as template .Sensitivity test showed that the RT-PCR method can detect as low as 0.69 pg/μL GDVII cDNA.There were no objective band amplified when encephalomyocarditis virus , lymphocytic choriomeningitis virus , Japanese B encephalitis virus , murine norovirus and normal mouse brain tissue were used as case-control .All infected mice showed symptom of different degrees such as depression and hind limb paralysis the 3th day postinoculation and two of infected mice died the 5th day postinoculation.Tissues such as heart, liver, spleen, lung, kidney, brain, cecal contents and serum were collected and tested for TMEV .All the brain samples were detected positive for GDVII and other tissues were all negative;The 100 cecal contents samples were tested and all were negative . Conclusions RT-PCR for TMEV GDVII strain can detect virus infection in mouse tissues efficiently and can be used as a powerful supplement for the national standard of lab animal .
9.The signal transduction pathway of rats with liver fibrosis regulated by leptin and interfering effects of mistletoe alkali
Xia MENG ; Xuecong WANG ; Ping FENG ; Jing LU ; Xuejiang WANG
Chinese Journal of Comparative Medicine 2015;(10):1-6
Objective To investigate the signal transduction pathway mechanisms of rats with liver fibrosis regulated by leptin and interfering effects of mistletoe alkali .Methods The hepatic fibrosis in rats model was established by injecting carbon tetrachloride .Forty-five SD rats were randomly divided into normal group ,model group and therapeutic group.All rats except rats in normal group were intraperitoneally injected with 40%carbon tetrachloride in peanut oil with a dose of 2.0 mL/100g according to the body weight twice a week for 8 weeks.Then, the therapeutic group was given mistletoe alkali (8g/(kg· d)) for 8 weeks via gastrogavage.Rats in normal and model group were served with distilled water at the same time.At the end of the 16th week, blood and tissue specimens were taken from all the rats .The influence of mistletoe alkali on liver morphology in liver fibrosis rat model was reviewed by HE and Masson staining .The effects of mistletoe alkali on the expression of Leptin and its receptor ( OB-Rb ) in HSC in fibrosis rat model were determined by immunohistochemistry (IH).The expression of JAK2, STAT3 and the activity of phospho -JAK2, phospho-STAT3 were detected by Western blotting analysis .Results The degree of fibrosis of the model group was more severe than the normal group and the treatment group , which suggested that mistletoe alkali can reverse liver fibrosis in rats . Immunohistochemical staining showed that mistletoe alkali reduced the hepatic expression of leptin and OB -Rb in rats with liver fibrosis in comparison with their expression in the model group .Compared with the normal group , the expression of JAK2 and STAT3 increased in the model group .However, the expression of JAK2 and STAT3 decreased in the medication groups compared with the model group .Conclusion Mistletoe alkali can effectively ameliorate liver fibrosis in rats possibly through inhibiting hepatic leptin and its receptor expressions , which through inhibiting hepatic leptin and its receptor expressions , thus inhibit the JAK2/STAT3 signaling pathway .
10.A method of tracheal intubation in mice mediated by optical fiber
Chinese Journal of Comparative Medicine 2015;(10):69-70
Objective To establish a stable and simple method of tracheal intubation in rodents and to establish various models that needs of endotracheal intubation .Method Sixty Kunming mice anesthetized by midazlam and ketamin, were intubated by 22 GA arterial puncture needle casing with optical fiber .Then connect the casing and ventilator and make sure the casing has been into the trachea .Result 60 mice were all successfully intubated by casing with optical fiber with no throat mucosa bleeding, edema.Glottis and trachea don’t present complications such as tracheal stenosis 7d later.Conclusion Fiber mediated intubation is a simple and useful method .It has a success rate with low incidence of complications .