Abstract:Objective To observe the NTBC dependence of Fah-knockout mice and study the biological characteristics in order to use the model more effectively. Methods Examine the progressive changes in body weight, survival time, liver pathology and serological markers after the NTBC withdrawal. Results After removing of NTBC, Fah-knockout mice lost their body weight gradually, and finally died in 5 to 7 weeks, along with increased serum ALT, AST levels and deformation of the hepatocytes. Conclusions Fah-knockout mice have a strong drug dependence of NTBC and could be the ideal model to hereditary tyrosinemia type I and other liver injury.

Abstract:Objective To get the gene encoding extracellular domains of gB protein of B virus and analyze its expression in the eukaryocyte cell. Methods synthesizing gene fragment encoding extracellular domains of gB protein of B virus was by using synthesis gene, then digested with the restriction endonucleases BamHⅠand NotⅠand inserted into eukaryotic expressing vector pEGFP-N3. pEGFP-N3-GB_合 was transfected into 293 cells. After protein extraction, the expression of gene was detcted by western blotting, and the cellular localization of the gene was analyzed by immunofluorescence and laser scanning confocal microscopy. Results pEGFP-N3-GB_合 were expressed in 293 cells and on the cell membrane. Conclusion eukaryotic expressing system can produce specific antigen recombination protein of B virus gB protein and express on the cell membrane.

Abstract:Objective To construct and identify retroviral-mediated short hairpin RNA(shRNA) expression vectors of ERβ419, and explore ERβ419 unknown biological function in beagles in future. Methods To screen out the most effective gene silencing sequence of beagle ERβ419 mRNA using qRT-PCR and Western Blot assays, imitate beagle estrogen target cells. Results qRT-PCR results showed,ERβ419-shRNA1(P <0.01)and ERβ419-shRNA3(P<0.01)differed significantly, Western Blot result as same as qRT-PCR,ERβ419-shRNA3 is the best choice. Conclusion Beagles ERβ419-shRNA3 retrain most effectively target gene repression. It is applied to explore ERβ419 unknown biological function in beagles reproductive system, and to prevent and treat beagles reproductive function diseases.

Abstract:Objective To quantitative the changing information of estrogen receptor β gene which was in tissue and organ of sex gland during oestrus and dioestrus of Beagles,and to show the different expression situation of hypothalamus-pituitary-gonad axis during oestrus and dioestrus, and providing the basic of theory to research deeply the mechanism of heat of Beagles. Methods As the key gene in regulation reproduction, ERβ gene is located in hypothalamus-pituitary-gonad axis, so using Beagles which was in oestrus and dioestrus, and extract the RNA from hypothalamus、pituitary、ovary and uterus respectively,after reverse transcription we detected the expression of ERβ gene by real-time quantitative PCR. Results The expression of ERβ gene mRNA from ovary、uterus、pituitary、hypothalamus of Beagles which was in dioestrus was 0.35 times, 0.17 times, 0.44 times and 0.43 times than the expression of ERβ gene mRNA from ovary, uterus, pituitary, hypothalamus of Beagles which was in oestrus. Conclusion The expression of ERβ gene was up-regulation in hypothalamus-pituitary-ovary axis of Beagles which was in oestrus.

Abstract:Objective To Identify B-cell epitopes on monkey B virus envelope protein gD. Methods Base on bioinformatics software, secondary structure, hydrophilicity, surface accessibility, antigenic index and flexibility of monkey B virus envelope protein gD was analysed and some potential peptide epitopes were forecasted. Then, the interactions between synthetic peptides and BV positive sera were detected by Enzyme-linked immunosorbent assay (ELISA). At last, The sensitivity and specificity of synthetic peptides was evaluated used 20 samples of Standard sera by ELISA. Result Seven epitopes were forecasted by Bioinformatics analysis. Four synthetic peptides, sequence as ⁴⁶LPPLEQKTD⁵⁴, ¹⁰⁶RGAPEATRSDA¹¹⁶, ²⁹¹PELAPEERGTSRTPGD³⁰⁶ and ³⁶¹AVYLVRRRGR³⁷⁰could be reacted with positive sera pool. The sensitivity of 4 synthetic peptides changed form 40% to 70% and the specificity were 100% for 4 synthetic peptides. Conclusion There are at least four linear epitope on B virus gD protein.

Abstract:Objective To find out and clone the genes that can influence different willis circle in Meriones unguiculatus, the genomic library has been made. Methods 96 clean Meriones unguiculatus were dissected according to observe the differences of willis circle, and the mixed DNA was extracted from different blood vessels. The genomic library construction has been done by using pCC1FOS vector, following by CopyControl Fosmid Lib manual. The copy numbers, recombined segment size and recombination fraction of the library were measured. Results Genomic library of different Willis circle in Meriones unguiculatus was successfully made. The capacity of the library was 1700 copies. The segment size was 36kb, and the recombination fraction was 93%. Conclusion The library was the good beginning for the following steps which were gene cloning and genescreen.

Abstract:Objective To screen the splicing isoforms of estrogen receptor β in the Beagle hypothalamic-pituitary-gonadal axis. Methods For ERβ mRNA CDS sequence of eight exons, primers were designed confined to the CDS sequences of two sequential exons. Beagle hypothalamus, pituitary, ovary and uterus tissue cDNA were used as template, and corresponding sequences were amplified by PCR. PCR products were sequenced and aligned in the NCBI web site. The correct gene was then analyzed with DNAMAN comparative analysis software and handwork checking up, thus got the ERβ splicing isoforms of Beagle. Results Four beagle ER beta splicing isomers were obtained: exon 4 complete skipping ER β isomer (300 bp missing), two kind of Beagle ERβ isoforms with partial exon 4 and partial exon 5 complicated missing (isoform Ⅰ334 bp missing and isoform Ⅱ 265 bp missing), and exon 7 complete missing ERβ splicing isoforms (181 bp missing). Exon 4 complete skipping and exon 7 complete missing isomers had been obtained full length coding sequence, and the other two splicing isomers were partial coding sequence. Conclusion This project gained four ER β splicing isomers of Beagle, and that will lay an important foundation for further study of their roles in the Beagle reproductive regulation mechanism.

Abstract:Objective To investigate if there are differences in hematological indexes between BALB/c mutant curly mice and normal BALB/c mice.Methods 6 week old BALB/c mutant curly and normal BALB/c mice of each 20(male and female)were selected,and 8 blood routine indexes and 15 serum electrolytes and biochemical indexes were detected using automatic blood cell analyzer and automatic biochemical detection,and the results of two groups were compared.Results The sex and group results of WBC,MCV,PLT and MCHC between BALB/c mutant curly mice and normal mice have significant differences(P<0.05,P<0.01);the sex and group results of Na⁺、K⁺ and Cl-between BALB/c mutant curly mice and normal mice have significant differences(P<0.05,P<0.01);the sex and group results of ALT、GLU and TG between BALB/c mutant curly mice and normal mice have significant differences(P<0.05,P<0.01).Conclusion Some hematological indexes between BALB/c mutant curly and normal BALB/c mice are different clearly,and these results will provide the theoretical references for researching and using mutant curly mice model.

Abstract:Objective To clone and sequence the dystonin variant X1 gene of Cricetulusbarabensis and the albino mutant Cricetulusbarabensis so as to find out the difference of encoding arear of the muscular ribosome between Cricetulusbarabensis and the albino mutant Cricetulusbarabensis. Methods According to the same type of abnormal muscle tone protein of the mice and rats,we designed 6 pairs of primers, and got their cDNA genes from skins of the Cricetulusbarabensis and the albino mutant Cricetulusbarabensis by RT-PCR amplification,then cloned and sequenced. Results Sequence alignment showed 17 variances in coding areas, and 24 in amino acid, but no in key nucleic acid and protein. Conclusion The variances in coding areas will not lead to the albino, and its mechanism requires further investigation.

Abstract:Objective Biodegradable is validated by the New Zealand rabbit femoral puncture mesh porous seepage prevention performance of the balloon. Methods The experiment was divided into the experimental group and control group, by piercing the New Zealand rabbit left leg near the femur, establish channel introduced after bone drill, fracture model is established, degradable mesh implant microporous balloon and/or bone cement. Observe and record the operation process of each New Zealand rabbit blood oxygen saturation change and cases of bone cement occur breakup and shortness of breath and groupings. Results Surgery under general anesthesia, bone cement injection process smoothly, balloon group of intraoperative breathing smooth, blood oxygen saturation has no obvious change; CPC group of blood oxygen saturation in the cement perfusion and perfusion after 81.63+/-32.02, 32.02+/-32.26, respectively, compared with perfusion 96.67+/-1.71 in front of the difference is statistically significant, of which 3 cases died in 5 cases, shortness of breath, direct infusion of bone cement is more likely to lead to pulmonary embolism. Conclusions Further confirmed that the degradable mesh porous balloon seepage prevention of CPC and the breakup of the effect is obvious, comply with the design requirements.

Abstract:Objective To compare the rat and mice intestine slides quality fixed with three different fixatives and embeded by paraffin. Methods Three kinds of conventional fixatives were selected for the test, they are 10% neutral buffered formalin,Bouin's solution and modified Davidson's solution. SD rat's and KM mice's intestinal tissues were fixed with the three different fixatives, then embeded, sliced and stained with HE as routine rut. To observe the slides quality under microscope. Results Sheding and necrosis of intestine villus epithelia were observed in slides made from formalin fixed intestines, which is speculated due to the fixations. The phenomenon was not observed in Bouin's solution and modified Davidson's solution fixed tissues. And the tissue structure were more fine and clear in them. Conclusions The rat and mice intestine slides made from tissues fixed by Bouin's solution and modified Davidson's solution are better than from the 10% neutral buffered formalin fixed ones.

Abstract:Objective To establish ectopic colon tumor model in GFP nude, make a comparative study of the differences between tumor morphology, biological characteristics and pathology with ectopic colon tumor model in BALB/c nude. Methods Use KM12SM colon cancer cells of the logarithmic growth phase seeded in right axillary subcutaneous on the GFP nude and BALB/c nude. Observed time of tumor formation, tumor volume, pathological conditions and other. Results The average time of tumor formation in GFP nude was 4~5 d, BALB/c nude was 6~8 d; The first 20 days after inoculation, tumor volume of BALB/c nude group was about(3463.43±341.39)mm³, GFP nude was about(4923.46±521.72)mm³. There are significant differences. Tumor cells growth activity, have relatively uniform size, some areas visible glandular formation on the microscope. Conclusion By comparison study, the GFP nude have certain advantages on the time of tumor formation, quality and other aspects in colon tumor model than the BALB/c nude. Provide more desirable animal model for investigating the mechanism of colon cancer lymph node metastasis, diagnosis and treatment.

Abstract:All organisms regulate their life activities through the biological clock, which makes a variety of activities regular. For example, many physiological activities such as sleep-wake cycle, temperature, heart rate, blood pressure, endocrine and metabolic activity of the kidney and liver are subject to the regulation of circadian rhythms, that is to say they are all under the control of circadian pacemaker. Physiological activity of cell cultured in vitro also possess rhythms.This paper conducts a brief overview of biological clock of cell cultured in vitro and analyzes the molecular mechanism of the biological clock of the neurons and peripheral tissue cell as well as the existing problems, which provide reference for comprehensive interpretation of the molecular mechanism of biological clock.

Abstract:Insulin is a hormone produced by islet β cells, which plays an important role in regulating cell division, cell differentiation, cell growth and regulating blood glucose levels. Decreased insulin and islet β cell dysfunction are vital fundamental characteristics of diabetes. There are many transcription factors involved in the development and maintaining of islet β cells. This review focuses on the transcription factors about β cell development.

Abstract:Objective To establish a showroom for experimental animal specimens, provide a shared platform for educational and scientific research of experimental animal. Methods ①The number of 140 animals which to be executed after normal experiment, using the technology of vascular perfusion, boiling peel, animal taxidermy and gross anatomy to make skeletal specimen, cast specimen, fur specimen, anatomical specimen,respectively; ②classification and exhibition. Results ①280 specimens were made, a showroom for experimental animal specimens was established; ②883 images were collected, digital database of specimens was established. Conclusion Showroom for experimental animal was a necessary place for students to integrate theory with practice, specimens can be lucid, vivid and convenient showed to research workers.

Abstract:This paper introduces the structure and the equipment of special artificial environmental and experimental chamber and its basic operating requirements. In compliance with the national standard and safety management, the safety, effectiveness and controllability of the chamber are described.