Abstract:Objective To screen for, and identify more-effective microsatellite loci for genetic quality analysisand enrichment of the genetic data in Mongolian gerbils using genomics. Methods Altogether, 357 repetitive sequences inaccordance with the microsatellite criteria were selected from whole-genome sequencing data of Mongolian gerbils. Thecorresponding primers were designed and synthesized based on the flanking sequences of each locus. The genomic DNA ofthe Mongolian gerbils was extracted and polymerase chain reaction (PCR)-amplified with their corresponding primers. Afteroptimizing the primer sequences, the PCR condition, and the gel electrophoresis experiments, the most efficient loci forsuccessful amplification were verified and optimized in various inbred lines and an outbred group of Mongolian gerbils. Theloci were also screened to identify those suitable for genetic quality analysis. Results Among the 357 microsatellitesequences, 135 loci primers were successfully amplified with the PCR product. Ten of them could be used to distinguish twoinbred strains of Mongolian gerbils, including the cerebral ischemia model and the diabetes model. Also, 12 gerbils fromthe outbred group had 23 loci that exhibited polymorphism. Conclusions In all, 135 microsatellite loci of gerbils aresuccessfully screened, some of which can be used for genetic quality analysis in both inbred and outbred groups of Mongolian gerbils.

Abstract:Objective To screen and validate aging-associated DNA methylation dysregulation and to establishconvenient, sensitive, accurate assays for detecting and screening substances capable of reversing dysregulated DNAmethylation associated with the aging process. Methods The Gene Expression Omnibus (GEO) database was used toscreen and validate DNA methylation alterations in the human aging process. A methylation qPCR detection system wasdeveloped to verify the correlation between DNA methylation levels of target genes and human age. Cancer cells were treatedwith a set of natural extracts/ substances that had anti-aging properties to detect their effects on DNA methylation levels oftarget gene and phenotypic changes. Results A methylation detection system for the secretagogin (SCGN) and integrinsubunit alpha 2b (ITGA2B) genes was established. Methylation levels of SCGN and ITGA2B genes were found to becorrelated with human age, which was verified in 53 independent samples. Epigallocatechin gallate (EGCG) and gallic acid(GA) inhibited the tumor cell survival and reversed SCGN and ITGA2B methylation dysregulation associated with aging.Conclusions Measurements of SCGN and ITGA2B methylation level changes may be used to monitor the aging process inhumans. They may also be used to provide experimental evidence for the development of anti-aging drugs or products based on DNA methylation regulation mechanisms.

Abstract:Objective To evaluate cardiac structure and function in rats with acute ischemic myocardial injury byultrasound cardiogram. Methods SD rats were randomly divided into three groups: sham group, model group withischemic myocardial injury, and a diltiazem treatment group (10 mg/ kg). Myocardial ischemic injury was induced byligating the left anterior descending coronary artery. Left ventricular structure and function were measured by ultrasoundcardiogram after 15 d. The parameters included EF, FS, SV, CO, LVIDd, LVIDs, LVPWd, LVPWs, LVAWd andLVAWs were assessed. The serum concentration of NT-proBNP was assessed using ELISA. Left ventricular dilation wasanalyzed through Evans and TTC staining. HE staining was used for the pathological examination of myocardial structure.Results Compared with the sham group, the degree of left ventricular dilation, the serum levels of NT-proBNP, andLVIDd and LVIDs were significantly decreased in the model group, but LVPWd, LVPWs, LVAWd and LVAWs wereincreased. Diltiazem treatment decreased LVIDd and LVIDs, and increased LVAWd, LVAWs, LVPWd, and LVPWs,compared with the model group, as assessed using an ultrasound cardiogram analysis. The diltiazem group also showed asignificant decrease in serum NT-proBNP levels, and left ventricular dilation compared with the model group. ConclusionsAn ultrasound cardiogram analysis is suitable as the primary method for evaluating cardiac structure and function in small laboratory animals with angiocardiopathies.

Abstract:Objective To investigate the changes of peripheral blood and immune cell phenotypes between agedand young C57BL/6J mice. Methods Peripheral blood cell count, immune organ coefficient and peripheral blood immunecell phenotype in aged C57BL/6J mice (19 months) and 2-month-old C57BL/6J mice used as control adult mice wereanalyzed. Results Compared with young mice, the aged mice had an increased peripheral blood platelet count, increasedproportion of neutrophils, and a decreased proportion of lymphocytes ( P <0. 001). The thymus coefficient was significantlyreduced ( P <0. 001). The percentage of CD4+ cells in the peripheral blood immune cell typing was obviously reduced ( P <0. 001). Conclusions The blood cells in aged mice are differentially skewed, and helper T-cell numbers declined. Thechanges are similar to those in the aging human. This study provides basic data for research on immuno-senescence.

Abstract:Objective To explore the role of CCR5 gene in the migration of myeloid-derived suppressor cells(MDSCs) into the mouse liver cancer microenvironment. Methods A mouse model of orthotopic liver cancer wasestablished and changes in the MDSCs were detected by flow cytometry. To observe the effect of mouse liver cancer cell H22(T-cell) conditional medium (TCM) on the migration of MDSCs, these cells were sorted and pretreated with 100 nmol/ LCCR5 inhibitor. Finally, DiR-labeled MDSCs were injected into the tail vein of mice with in situ hepatocarcinoma to observethe effect of the CCR5 inhibitor on the migration of MDSCs in vivo. Results The expression of MDSCs in bone marrow,spleen, and liver was significantly higher in the mouse model of orthotopic liver cancer than that in normal mice. The purityof MDSCs was 94. 5%, and MDSCs expressed CCR5. Cell migration result showed that CCR5 inhibitor reduced therecruitment of MDSCs by TCM. The in vivo imaging analysis showed that the ability of MDSCs to migrate to the spleen andliver in vivo was weakened after treatment with CCR5 inhibitor. Conclusions CCR5-mediated MDSCs accumulate in the mouse liver cancer microenvironment. Inhibition of CCR5 in MDSCs may provide a new idea for cancer treatment.

Abstract:Objective The spermatogenesis- and oogenesis-specific bHLH transcription factor-1 (SOHLH1) andSRY box (SOX30) are transcription factors related to spermatogenesis. Sohlh1 and Sox30 knockout male mice lose fertility.At 7 days after birth, the Sohlh1 knockout male mouse DNA chip exhibits significant down-regulation of Sox30 expression.This study explored the role of the early development-related transcription factor SOHLH1 in the transcriptional regulation ofthe key gene Sox30 during late spermatogenesis. Methods The Sox30 promoter luciferase expression plasmid wasconstructed, and then together with the Sohlh1 eukaryotic expression plasmid was transiently co-transfected. Thefluorescence value was then detected by a luciferase activity assay. The primary activation site was screened using achromatin immunoprecipitation assay to detect the binding of SOHLH1 to the Sox30 promoter DNA sequence. Results Theluciferase activity was higher in the experimental group co-transfected with the Sohlh1 eukaryotic expression plasmid and theSox30 promoter region luciferase plasmid than in the control group. SOHLH1 binding to specific sites was strongest on theSox30 promoter binding sites at -489 bp. Conclusions The early important transcription factor SOHLH1 can directlyactivate acrosome-forming gene Sox30 transcription, with -489 bp (CAGGTG) as the main specific binding site. Thisaction enriches the regulation of delayed translation expression during spermatogenesis. The study thus provides a preliminary exploration of the regulatory mechanism.

Abstract:Objective To investigate the difference between BALB/ c mutant curly mice and control BALB/ cmice in terms of natural healing of skin injuries. Methods Ten 6-week-old BALB/ c mutant curly mice and 10 controlBALB/ c mice were selected. Two full-thickness skin wounds with a diameter of 0. 5 cm were created on the back of themice. A skin injury model was used. Skin healing was observed at 3, 7, 10, and 14 days after injury, and changes in theskin damage area and the wound healing rate were measured. Animals were sacrificed at 3, 7, 10, and 14 days after injury.HE staining and Sirius staining were performed to observe pathological changes in the skin during healing. Results Theskin wounds healed faster in the mutant curly mice at 3 and 7 days after wound healing. The skin wound healing rate of thecurly mice at 3 and 7 days after injury was significantly higher than that of the control mice ( P < 0. 05), but there was nosignificant difference between the two groups at 10 and 14 days after injury. The pathological examination showed that thecollagen fibers of the mutant curly haired mice increased significantly 3 days after injury, and there was a small amount ofinflammatory cell infiltration, and capillary dilation and congestion. Collagen grew rapidly 7 days after injury, levels ofgranulation tissue increased significantly, and some epidermis regenerated and grew. The Sirius scarlet staining resultshowed that collagen fibers and granulation tissue were present from the bottom layer of the injury, to repair the damagedarea, until the epidermis and hair follicles grew out from 3 to 14 days in the curly-haired mice. Conclusions BALB/ cmutant curly mice have better healing ability than control mice in the early stage of skin trauma, which provides a theoretical reference for the application of BALB/ c mutant mice in skin wound healing models in the future.

Abstract:Objective To study the protective effect of trigonelline on testicular injury induced by heat stress inmice. Methods A total of 120 ICR male mice were randomly divided into a control group, model group, and trigonellinegroups (25 mg/ kg, 50 mg/ kg, 100 mg/ kg), with 24 rats in each group. The lower abdomen of the mice in the model andtrigonelline groups was placed in a constant temperature water bath at 43℃ for 15 min to induce heat stress, and then thetrigonelline group was treated using trigonelline through intragastric gavage for 14 days. The mice in the control group andthe model group were treated using saline for 14 days. Animals were sacrificed at 1, 7, 10, and 14 days after heatstimulation. The genital organ index, a sperm count, and HE staining were used to observe changes in testicular tissuestructure and for a comparative analysis. Results Compared with the control group, the genital organ coefficient and thenumber of epididymal sperm in the model group were significantly decreased ( P < 0. 05). A histopathological analysisshowed obvious testicular injury in the mice. Trigonelline treatment increased the genital organ coefficient and the number ofspermatozoa in the epididymis, compared with the model group ( P < 0. 05). Histopathology showed that trigonellinetreatment significantly reduced heat stress-induced injury in the mice. Conclusions Trigonelline can significantly improvethe testicular coefficient and reduce heat stress-induced damage in testis tissue in mice, and it has a certain antagonistic effect.

Abstract:Objective Osteoarthritis (OA) is the most common type of degenerative chronic joint disease, butthe exact genetic mechanisms are still unclear. The aim of this study was to analyze the gene expression profile in an OAcell model detected using a gene chip, and to provide a biological basis for the pathogenesis of OA. Methods Primarymouse chondrocytes were isolated using trypsin combined with collagenase, and the cells were incubated with 50 ng/ mLTNF-α for 24 h. Total RNA was extracted from the harvested cells for gene chip detection to identify differentially expressedgenes (DEGs). A fold change (FC) greater than two and P<0. 01 were the conditions required for each DEG. Biologicalinformation software was used to conduct gene ontology (GO) and KEGG pathway annotation analysis of DEGs. Results After TNF-α treatment, a total of 8096 up-regulated DEGs and 6413 down-regulated DEGs were identified, including genesthat are known to be associated with OA, such as matrix metalloproteinase, inflammatory factors, and apoptosis-related andosteoblast-related genes. In addition, Olfml1 and other olfactomedin superfamily members, Nf1 and other previouslyunreported genes related to OA, were also found. In particular, abnormal expression of a large number of genes related tocytochrome C superfamily members was found, suggesting that mitochondria-related functional genes and signaling pathwaysmay be significantly associated with OA. Conclusions The changes in the gene expression profile in a TNF-α induced OAchondrocyte model at the level of the transcriptome are datected in this study, providing new insights into the pathogenesis of OA.

Abstract:Objective To establish a hyperlipidemic cynomolgus monkey model with human diseasecharacteristics, and to provide an ideal animal model for the development of lipid-lowering drugs, especially lipid-loweringbiopharmaceuticals. Methods The male cynomolgus monkeys in different treatment groups were gavaged with a formulatedsemi-liquid high fat/ high sugar dietary emulsion 6 days per week. Body weight (BW), body mass index (BMI), serumtotal cholesterol ( TC), triglyceride ( TG), high density lipoprotein cholesterol ( HDL-C), low density lipoproteincholesterol (LDL-C), and other indexes were monitored regularly, and changes in the blood lipid profiles were analyzed.Results Compared with the corresponding control group, by the third month of modeling, all six middle-aged and old malecynomolgus monkeys and five adult male cynomolgus monkeys in the treatment group were successfully modeled. By the 21stmonth, all male cynomolgus monkeys in the treatment group still maintained a relatively high level of blood lipid.Conclusions A hyperlipidemic cynomolgus monkey model is successfully created through quantitative gavage using a highsugar and high fat dietary emulsion for 3 months. The model demonstrates the characteristics of hypercholesterolemia andhigh-density lipoprotein cholesterolemia, and it can be used to study the pathogenesis of hyperlipidemia and for pre-clinical drug evaluations of pharmacodynamics.

Abstract:Objective To evaluate the effect of murine norovirus (MNV) infection on immune function in miceby assessing the changes in peripheral blood immune parameters. Methods KM, BALB/ c, BALB/ c-nu, C57BL/6, andNIH mice were selected. Each strain was divided into an MNV infection group and a control group. Anticoagulated bloodsamples were collected from the periocular vein plexus before infection (day 0) and at 7, 14, 21, and 28 d post-infection.Flow cytometry was used to determine the percentage of total T cells, CD4⁺ T cells, CD8⁺ T cells, total B cells, NK cellsand NK-T cells in the total lymphocytes and the levels of 13 cytokines such as interferon and interleukin, and chemokines.To compare the differences in the mean values of each index after infection, the changes at each time point were analyzed.Results Compared with the control group, the level of total T cells and CD4⁺ T cells in the infected group of KM micesignificantly increased ( P <0. 01), CD4/ CD8 significantly increased ( P <0. 05), and the total B cell and CXCL10 contentsignificantly decreased ( P <0. 01). The level of CD8⁺ T cells in infected BALB/ c mice significantly decreased ( P <0. 05),and IFN-γ levels significantly increased ( P <0. 01). There was no significant difference between the levels of immune cellsin infected and control BALB/ c-nu mice, although in the infected BALB/ c-nu mice TNF-α, CXCL1 and CXCL10significantly increased ( P <0. 01), CCL2 significantly increased ( P < 0. 05), and IFN-β significantly decreased ( P <0. 05). CD4⁺( P <0. 05) and CD8⁺ T cells ( P <0. 01) in the infected C57BL/6 mice significantly decreased, total B cellsand IL-10 significantly increased ( P <0. 05), and CCL5 and GM-CSF significantly decreased ( P <0. 05). Total T cells andCD8⁺ T cells in the infected NIH mice significantly increased ( P <0. 01), CD4/ CD8 significantly decreased ( P <0. 05),and total B cells significantly decreased ( P <0. 01), while there were no significant differences in cytokine levels. In termsof statistically significant changes in immune indicators, half of all immune indicators showed significant differences on the7th day of infection, and most still demonstrated significant differences on the 28th day. Conclusions MNV infectionaffects cellular and humoral immune responses in otherwise healthy mice, and different mouse strains demonstrate differenteffects. Researchers should select MNV-negative mice in immunologically relevant animal experiments to avoid bias in the experimental results.

Abstract:Objective To study the influence of acute exercise on rat myocardial microRNA-30a and autophagyrelatedgene expression, and investigate the possible mechanism through which microRNA-30a regulates myocardial autophagyduring different recovery periods after exercise. Methods SD rats were randomly divided into a control group (Con), a 0 hafter exercise group (E0), a 3 h after exercise group (E3), and a 12 h after exercise group (E12). Real-time fluorescencequantitative polymerase chain reaction was used to detect the expression of ACE mRNA, microRNA-30a, Beclin-1 mRNA,and LC3 mRNA in the left ventricular myocardium. Results Compared with the control group, in the E0 group, ACE mRNAand microRNA-30a gene expression in the left ventricular muscle was significantly increased ( P <0. 05), and Beclin-1 mRNAand LC3 mRNA expression also significantly increased ( P <0. 05). This was recovered to resting state approximately 3 h afterexercise. Conclusions Acute exercise can significantly improve myocardial ACE mRNA, microRNA-30a, Beclin-1 mRNA and LC3 mRNA expression. This transient increase may be caused by exercise-induced stress.

Abstract:Animal biosafety level-2 (ABSL-2) facilities are used for infected animal experiments. The use of suchfacilities is very high in many scientific research institutions. The efficient management of the facility is a precondition andassurance of animal experiments. In the current article, the operation management experiences of staff in ABSL-2 facilitieswere recorded, such as in the Chinese Center for Disease Control and prevention, and the key points for daily management in such facilities are highlighted to provide a valuable reference.

Abstract:Objective To determine an effective anticoagulant treatment program by the management ofanticoagulants after the implantation of a new left ventricular assist device (LVAD) in sheep. Methods Six male healthysheep weight 80±5 kg were implanted with the same type of LVAD. Blood was collected once a day in the first two weeksand once a week thereafter. The sheep were observed and evaluated for 10 weeks. Results The physiological state of thesix experimental animals: the international normalized ratio (INR) was 1. 265±0. 146; activated partial thromboplastintime (APTT) was 41. 017±12. 530 s; lactate dehydrogenase (LDH) level was 411±119 U/ L. One to three days afteroperation, the control of INR and APTT by heparin and warfarin were 1. 4 and 1. 7 times higher. Heparin was stopped whenthe animal could eat again. INR was monitored continuously and the warfarin dosage was adjusted. Two weeks afteroperation, warfarin 7. 96±5. 45 mg/ d was used and the INR was 1. 89±0. 53. Ten weeks after operation, warfarin 11. 44±7. 80 mg/ d was used, the INR was 2. 01±0. 91 and the level of LDH was 509±92 U/ L. During the study, no thrombuscaused by the pump was found, and no gastrointestinal bleeding was found in experimental animals. Conclusions It isnecessary to apply heparin early after the operation. Under the premise of not changing the diet structure, the anticoagulanttreatment standard for each individual is established within 2 weeks. The use of warfarin only can achieve the expected anticoagulant effect.

Abstract:Liver fibrosis is caused due to excessive deposition of diffuse extracellular matrix in the liver duringrepair of liver injury caused by various pathogenic factors. In recent years, an increasing body of evidence has suggestedthat the phosphatase and tensin homology deleted on chromosome 10 (PTEN) is involved in the development of liver fibrosis. We therefore reviewed the regulatory mechanism of PTEN in liver fibrosis.

Abstract:The tumor microenvironment was once considered to have little effect on tumor growth and invasion.However, it has now been recognized that it plays an indispensable role in tumor development. One of the commoncharacteristics of tumors is hypoxia, which is also one of the representative characteristics of the tumor microenvironment.Hypoxia leads to an increase in extracellular hydrogen ion concentrations, a decrease in pH (acidosis), and the formationof a specific acidic microenvironment. The acidic microenvironment plays an important role not only in the proliferation,migration, invasion, metastasis and treatment of cancer cells, but also in the function of immune cells, vascular cells andother stromal cells. However, the molecular mechanism through which tumor cells perceive and respond to acidic pH intheir microenvironment remains unclear. Previous studies have shown that some receptors and ion channel families can helpcells to perceive acidosis. As a G-protein coupled receptor (GPCR), ovarian cancer G-protein coupled receptor 1 (OGR1)is activated by acidosis and mediates several downstream G-protein signal transduction pathways. It can not only regulatemetastasis and proliferation in cancer cells, but also plays an important role in the function of immune cells, inflammationand angiogenesis. In the current review, the effects of OGR1 in tumorigenesis, the tumor microenvironment, the immuneresponse and angiogenesis are discussed. The development of small molecule regulators targeting GPCRs has become an areaof active research in recent years. Studying the interaction between OGR1 and the tumor microenvironment will provide new strategies for cancer treatment and chemoprevention.

Abstract:Neural tube defects(NTDs)is a common neurological disease in the world, with an occurrence of about1/ 1000. The pathogenesis of this disease is complicated, which is determined by genetic factors and environmental factors.Till now, there is no effective treatment, however supplying folic acid can reduce the incidence of neonatal NTDs. Presentstudies focused mainly on zebrafish, Xenopus and mouse experiments, and found that the planar cell polarity andconvergent extension play an important role in neural tube closure. To better understand the function and molecularmechanism of PCP pathway in the process of neural tube closure, this review summarized the current progress in research ofthe disease and the relationship between the disease and three genes: SHROOM3, VANGL2 and WNT5A, in addition summarized the animal models of NTDs.

Abstract:The exploration of human disease using animal disease models is an important means to study themechanisms underlying disease occurrence and the clinical effects of drugs. At present, various animal models of renalfailure have been developed, but there are few comparative studies of such models. Based on a rat kidney failure model as abreakthrough point, the current paper analyzes and compares rat pathological models of chronic renal failure and acute renalfailure. This includes assessments of the methods used to create the models, the value of the model itself and its practicalresearch applications in the clinical research of the pathogenesis of renal failure, and drug research and development. The most appropriate animal experimental model for specific applications is discussed.

Abstract:Dilated cardiomyopathy (DCM) is one of the most common causes of heart failure. The etiology of DCMis complex and its mechanisms are still not clear. As a severe disease threatening human health, DCM has been a populartopic of clinical and laboratory research. An appropriate animal model is an essential prerequisite for the research. In thispaper, the animal models used in relevant literature both at home and abroad were reviewed and analyzed in order to provide reference for further research.

Abstract:Low-level laser therapy is increasingly being used in the medical field, and has apparent advantagessuch as non-invasive, non-toxic side effects compared with those associated with traditional surgery, drugs, and radiationtherapy. It has the positive effects of reducing inflammation, promoting wound healing, nourishing nerves, protectingmuscles, and improving body shape. An in-depth understanding of these mechanisms of action for low-level laser therapycan help to improve the efficacy of the therapy and broaden its range of applications. This article reviews the advances in the application and our understanding of the mechanisms of low-level laser therapy.

Abstract:Major depressive disorder is one of the most common and serious mental disorders and is characterizedby constant low spirits and emotional disorder. Establishing a suitable animal model of depression will benefit the study ofdepression-related problems. Here, we have reviewed the animal species, behavioral tests, modelling methods and theunderlying mechanisms in most widely used animal models of depression and new models in development. Rodents havebeen used in depression models for many years. Non-human primate animal models are under research and such animalsmay have similar behaviors as human beings. Different animal models of depression have similarities and differences interms of modelled behavior, endocrine and metabolic characteristics, social psychology, and genetics. The characteristics of different models should be considered when choosing one.

Abstract:This study recorded and arranged the data concerning the Chinese Laboratory Animal License InquirySystem, and analyzed the current situation and problems concerning laboratory animal license administration. There is a needto strengthen the propaganda and post-event supervision of laboratory animal license system administration. We have also torapidly develop and use new laboratory animal resources. This article provides a reference for the government decisionmaking,optimal allocation of laboratory animal resources and promote the healthy development of laboratory animal industry.