Naked mole rats are unique animals with long lifetime, anti-tumor properties, hypoxia tolerance, low metabolic rate, analgesia, tactile sensitiveness, poor eyesight and strong bone regeneration ability. Based on the basic bio-logical characteristics of naked mole rats, and the current research trends in the field of cancer, aging, hypoxia tolerance, as well as algesia, this review focuses on the application prospects of naked mole rats in biomedical research.

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.

Objective:To explore the way to separate and culture eutopic and ectopic endometrial glandular cells and their stromal cells,providing an in vitro cell model of endometriosis for study of its mechanism. Methods:Digestion,filtration and sedimentation were used to isolate and culture the eutopic and ectopic endometrial glandular cells and their stromal cells. The estrogen level was imatated to study the way of promoting cell growth. Morphological characters of eutopic and ectopic endometrial cells were examined using optical microscope. Results:The success rate of separation and culture of normal control endometrial glandular cells and its stromal cells was 91.7%(11/12);of eutopic endometrial glandular cells and its stromal cells of endometriosis was 93.8%(15/16);of etopic endometrial glandular cells and its stromal cells of endometriosis was 75.0%(12/16) . Conclusion:The cultured eutopic and ectopic endometrial cells is more like human body feature than the endometriosis model of animals. So the isolation and culture of eutopic and ectopic endometrial glandular cells and their stromal cells may serve as an in vitro experimental model.

This paper discusses application of multimedia and network technology in Laboratory Animal Science Teaching:We have set up multimedia courseware,made animal experiment video and built network course-learning system and a good teaching effect has been achieved.Then it analyzes the problems of multimedia and network technology in Laboratory Animal Science Teaching.

According to the principles of constructing evaluation index system in designed experiments,we formulated the content,designed the weights and established the evaluation index system in designed experiments based on analytical hierarchy process and Delphi.This system can assess students' cognition and attitudes,abilities of innovation,practice,teamwork and cooperation as well as scientific writing skills by providing objective and comprehensive evaluation criteria.

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.

Objective To detect the protection induced by HPV-58 L2 11-200 AA in animal, and analyze the relationship between antibody or neutralizing antibody titers and the protection generated by the immunizmg agent. Methods The peptide of HPV-58 L2 11-200 AA was expressed in E. coli and the mice were immunized with the peptide after purification and adsorption with aluminum adjuvant. The protection provided by different immunizing doses was detected in the mouse model against the challenge of the pseud-ovirions of human papiilomavirus types 58. The total antibodies and neutralizing antibody titers of serum were tested with ELISA and neutralization assay against HPV-58 pseudovirus, respectively. The total antibodies or neutralizing antibody titers that can protect the mouse from infection were analyzed. Results The mice can be protected from the challenge with HPV pseudovirus when the immunizing dose was 8 μg. The neutralizing antibody can not be detected in the immune serum by neutralization assay against pseudovirus. The total anti-body level has a corresponding relationship with the protection showed in mouse model. The results of total antibodies detected by ELISA showed that when the titer of total antibodies was ≥25 000, luminescent signal can not be detected and the mice can be protected from pseudovirus infection. Conclusion HPV-58 L2 11-200 AA peptide can protect mice from pseudovirus infection. L2 peptide has a promising perspective to be a candidate vaccine and the level of total antibodies in the immune serum can be used as a surrogate for the evaluation of protection against HPV infection.

On hundred cases of B—hepatitis were treated by self—formulatel B—hepatitis No.1 granules made of traditional drugs.A control group of100 cases were treat-ed with Jiaosanxian(scorched triple ingredients,con-sisiting of hawthorn,malt.and membrane of chickengizzard).Resultsshowed that,in the experimentedgroup,the lowering of transaminase,inhibition ofvirus duplieation and improvement of globulin ratioareall—superior to the control group.The resultswere also verified bylaborarory experiments.

Objective To investigate the effects of down-regulation of Beclin 1, which is an autophagy regulatory molecule, expression induced by RNA interference on the proliferation and apoptosis in skin fibroblasts of naked mole rat. Methods The expression levels of Beclin 1 were detected after starvation or H2 O2 treatment.The fibroblasts were transi-ently transfected with specific siRNA targeting Beclin 1 and then screened by real-time PCR and Western blot.Cell prolifer-ation and apoptosis were determined using CCK-8 detection kit and flow cytometry ( FCM ) .The expressions of related genes were detected by Western blot.Results The expression of Beclin 1 gene at mRNA and protein levels was signifi-cantly lower in fibroblasts of the naked mole rat.Starvation and H2 O2 treatment induced changes of the Beclin 1 expression. Inhibition of Beclin 1 gene expression can inhibit cell proliferation and induce early and late apoptosis.The protein levels of p53, BAX, Bcl2, LC3B, p-AKT and mTOR were reduced after transient transfection with Beclin 1-siRNA.Conclusions The expression of Beclin 1 in fibroblasts of naked mole rat are changed in response to starvation or H2 O2 stimulation.Inhi-bition of Beclin 1 gene expression can inhibit cell proliferation and induce apoptosis.Therefore, Beclin1 gene may play a regulatory role in autophagy, proliferation and apoptosis in the skin fibroblasts of naked mole rat.

Objective To construct plasmids and knock out HIF-1α gene expression in an naked mole rat skin fibroblasts(NSF)cell line using CRISPR/Cas9 genomic editing technology,to provide an in vitro cell model for studying the mechanism of hypoxia tolerance and the occurrence and development of hypoxia-related diseases in naked mole rats.Methods We designed four pairs of single guide RNA(sgRNA)sequences targeting exons 1～4 of the NSF HIF-1αgene and successfully constructed an expression plasmid.The plasmid with the optimal sgRNA was identified and transfected into 293T cells,and the supernatant was used for detecting the virus titer.Lentivirus particles carrying sgRNAs of HIF-1α were transfected into NSF cells which express Cas9 protein,based on a previous protocol.After transfection,fluorescence signals were observed under a fluorescence microscope,and HIF-1α expression in NSF cells was detected by Western Blot and T7 endonuclease 1(T7E1)analysis.Results Sanger sequencing showed that the designed sgRNA was successfully inserted into pX459 and pKLV2-U6-sgRNA2 vectors,demonstrating successful construction of a recombinant plasmid for transfection.T7E1 digestion successfully removed three bands and the target efficiency of sgRNA was 54％.Western Blot showed that the HIF-1α gene was successfully knocked out and its protein level was significantly reduced in NSF cells from naked mole rats(P=0.0019).There were no obvious morphological changes in HIF-1α-knockout cells under the microscope,and gene knockout had no obvious effect on cell proliferation.Conclusions We successfully constructed an HIF-1α-knockout cell line using CRISPR/Cas9 technology,to provide an experimental basis for further studies of the biological function of HIF-1α,as well as the mechanism of hypoxia tolerance in naked mole rats.The result also provide a theoretical foundation for the prevention and treatment of hypoxia-related diseases.