Two lectures coveting the management of laboratory mouse colonies werepresented in November at the laboratory animai medicine technieal trainingseminars sponsored by the Chinese Association for Laboratory Animal Sciences in Xi'an. Topics covered include troubleshooting mouse reproductiveperformance and managing genetically engineered colonies.The purpose of this article is to highlight the materials covered andto offer references for further reading. It is advised that references cited in this article be reviewed for a morecomprehensive coverage on the topics presented.Please also note that the principles for breeding practice and genetic management ofoutbred mice were not covered,but the author may be contacted for a list of suggested reading materials.

Mouse and vole embryos were allogeneically and xenogeneically transferred into pseudopregnant CD.1 and immunodeficient (seid)female mice,and we investigated the distribution of uterine leucocytes cells in the implantation sites on days 5,6,and 7 of pregnancy. Maerophages were evenly distributed in the endometrium on days 5-7.Neutrophils were rarely seen on days 5-7,but lymphocytes were found throughout the endometrium,often in groups associated with glands or the luminal epithelium.The number of uNK cells increased markedly at the mesometrial uriangle and the outer decidual area in the CD-1 uteri containing vole embryos;by contrast,seid uteri having vole embryos showed almost the same number as those having mouse embryos.Mast cells were present in large numbers at the myometrium,but rarely in the decidua in all types of pregnant uteri.Cells at the myometrium were more numerous in xenogeneic than in allogeneic transfer.Maay mast cells appeared in the inner decidua where xenogeneically transferred vole embryos were dead and aborted.These results suggest the possibility that uterine leucocytes mediate various immunological events in the mouse-vole interspesific pregnancies.

With increasing globalization and a trend towards international harmonization of standards for the care and use of animals in research and testing,there is a significant need to assist and support countries to develop training programs for laboratory animal veterinarians. Although formal educational opportunities for training laboratory animal veterinarians exist through well-established specialty colleges of laboratory animal medicine such as ACLAM,ECLAM,JCLAM,and KCLAM or through other professional organisations,such as the Federation of European Laboratory Animal Science Associations ( FELASA ),opportunities for participating in these programs are often limited to veterinarians in North America,Western Europe and specific regions of Asia. Creative thinking is required to develop cost-effective,practical,entry-level and advanced continuing education and applied training programs for veterinarians working in the field of laboratory animal medicine around the world.This paper will describe one potential solution for this issue,the use of a distance education program that provides theoretical information in a virtual classroom with applied training modules to deliver knowledge and practical skills to laboratory animal veterinarians.This type of program takes advantage of the online learning environment and can be an effective means to deliver training at the grassroots level to adult learners.

Objective To observe and analyze functional areas of the cerebral cortex in C57BL/6 mice of various ages.Methods Improved alkaline phosphatase staining was used to reveal the microvascular morphology of the cerebral cortex in C57BL/6 mice,including the motor cortex(primary and secondary motor cortex),sensory cortex(primary and secondary somatosensory cortex),visual cortex(primary and secondary visual cortex),and auditory cortex(primary and secondary auditory cortex),olfactory cortex(extrarhinal and entorhinal cortex).Images were captured under an OLYMPUS BX51 microscope with Image-Pro Plus 5.1 software.The microvascular length density(Lv),microvascular surface area density(Sv),and microvascular volume density(Vv)were analyzed by Image-Pro Plus 5.1 software.Results Expression of alkaline phosphatase was abundant in cerebral cortical microvessels of adult and elderly mice,and slightly expressed in juvenile mice,but not in lactating mice.Pial blood vessels enter the cortex in T shape,Y shape,large arc,and small arc four manners.Lv,Sv and Vv in different parts of the same aged mice showed a decreasing trend in motor,sensory,visual,auditory and olfactory cortexes,and the microvascular density of Lv,Sv and Vv in motor and sensory cortexes was statistically significant compared with the olfactory cortex(P<0.05).The vascular density in all functional areas in elderly mice was lower than that in adult mice,but no statistical significance was found(P>0.05).Conclusions The expression of alkaline phosphatase in microvessels in functional areas of the cerebral cortex in C57BL/6 mice increases with age and reached its peak value in adulthood.The microvascular architecture in the brain provides morphological parameters to establish cerebrovascular disease models.

Objective To investigate the expression of immunoglobulin γ-1 heavy chain constant region(IGHG1)in acute myeloid leukemia(AML)THP-1 cells and its influence on cell proliferation,apoptosis,and invasion via its regulation of the transforming growth factor β(TGF-β)/Smad pathway.Methods Bone marrow specimens from nine children with AML and eight children with fractures,human bone marrow stromal HS-5 cells,and human AML THP-1 and HL60 cells were used in the research.Western blot was used to detect IGHG1 protein expression.THP-1 cells were divided into a blank group(without any treatment),si-NC group,si-IGHG1-1 group,si-IGHG1-2 group,si-IGHG1-3 group,TGF-β group,si-IGHG1-1+TGF-β group,and si-IGHG1-1+TGF-β +LY364947 group.Cell proliferation was measured by CCK-8 method.Flow cytometry and Transwell experiment were performed to measure apoptosis and invasion.Western blot was used to detect the protein expression of IGHG1,TGF-β,p-Smad2 and p-Smad3 in each group of cells.Results Compared with the bone marrow of children with fractures,the bone marrow of children with AML(P<0.05)had significantly higher expression levels of IGHG1((0.24±0.03)vs(0.87±0.12)).Compared with HS-5 cells,human AML THP-1 and HL60 cells had significantly increased expression levels of IGHG1((0.89±0.14)(0.75±0.08)vs(0.21±0.02))(P<0.05).Compared with the blank group,the si-IGHG1-1 group of THP-1 cells showed significantly reduced OD450 values(24 h,48 h,72 h),invading cell numbers and protein expression of TGF-β,p-Smad2,p-Smad3 and their apoptosis rate was increased(P<0.05),while the corresponding indexes showed the opposite trend in the TGF-β group(P<0.05).TGF-β reversed the effects of silencing IGHG1 on the proliferation,apoptosis and invasion of THP-1 cells.Compared with the si-IGHG1-1+TGF-β group,the si-IGHG1-1+TGF-β+LY364947 group had significantly decreased TGF-β,p-Smad2,p-Smad3 and protein levels,OD450 values and cell invasion numbers and the apoptosis rate was increased(P<0.05).Conclusions IGHG1 is highly expressed in AML cells.Silencing the IGHG1 gene can inhibit the proliferation and invasion of AML cells and promote the apoptosis of AML cells.This mechanism may be related to the inhibition of the TGF-β/Smad pathway.

Objective To investigate the protective effect of geniposide against diabetic rats with skin ulcer and the mechanism.Methods Rats were divided into a normal group,model group,and geniposide subgroups(Gen(L):200 mg/kg;Gen(H):500 mg/kg).Diabetic rats were treated with normal saline or geniposide by intragastric administration(n=6).Treatments were administered once a day,and the wound healing and inflammation of each group were recorded every day.After 7 days of treatment for diabetic skin ulcers,the wound area,tissue sections,TUNEL staining and Western blot were used to quantitatively analyze changes in wound healing,apoptosis,and related regulatory protein expression.Results Compared with the model group,the group receiving orally administered geniposide(200 and 500 mg/kg)showed significantly improved wound healing and increased contraction of the injured area.In terms of skin wound apoptosis in diabetic rats,TUNEL-positive cells were significantly reduced in geniposide subgroups(P<0.05).Geniposide significantly inhibited skin inflammation and promoted wound repair,which may be related to promotion of PI3K and Akt phosphorylation.Conclusions Geniposide promoted skin wound repair in diabetic rats by inhibiting inflammatory responses and apoptosis.

Objective To investigate the effect of human amniotic epithelial cell(hAEC)transplantation on endometrium improvement and matrix metalloproteinase 8(MMP-8)and vascular endothelial growth factor(VEGF)expression in a rat model of uterine scaring.Methods The uterine scar model was established in rats that were randomly divided into model and transplantation groups with 18 rats in each group.The other 18 rats were used as the sham operation group.Rats in the transplantation group were injected with hAECs in the uterine scar,and rats in model and sham operation groups were administered the same amount of PBS.After 4 weeks,the uterine tissues of eight rats in each group were collected.Histomorphological changes and endometria fibrosis were observed by HE staining and Masson staining respectively,and the endometrial thickness and number of glands were measured.Endometrial growth and receptivity were evaluated by immunohistochemical staining of cytokeratin and integrin β3,respectively.mRNA expression of MMP-8 and VEGFA in endometrial tissues was measured by RT-qPCR.Western blot was used to measure MMP-8 and VEGFA protein expression.After 8 weeks,the remaining 10 rats in each group were used to assess gestational ability.Results The endometrial thickness,gland number,IOD value of keratin and integrin β3,relative mRNA and protein expression levels of MMP-8 and VEGFA,pregnancy rate and number of uterine embryos in model and transplantation groups were lower than those in sham operation group(P<0.05).The endometrial thickness,gland number,IOD value of keratin and integrin β3,relative mRNA and protein expression of MMP-8 and VEGFA,pregnancy rate and number of uterine embryos were higher than those in model group(P<0.05).Additionally,hAEC transplantation improved the pathological morphology of endometrial tissue in rats with uterine scaring and reduced the degree of endometrial fibrosis.Conclusions hAEC transplantation improves endometrial injury,reduces scar formation,improves endometrial receptivity,and enhances pregnancy function in model rats,which may be related to promotion of MMP-8 and VEGFA expression.

Objective To investigate the effects of Osteoking on hyperglycemia and regulating gut microbiota in db/db mice.Methods Wildtype mice were used as the control group,and db/db mice were randomly divided into model and Osteoking groups.After intragastric administration for 12 weeks,fasting blood glucose and serum glycosylated hemoglobin and insulin levels were measured,changes in intestinal microflora were determined,and functional pathways related to intestinal microflora in mice were predicted by 16S rDNA sequencing.Results Compared with the model group,Osteoking decreased fasting blood glucose(P<0.01),serum glycosylated hemoglobin(P<0.01),and the insulin resistance index(P<0.01),and increased insulin content(P<0.01)in db/db mice.Osteoking increased the abundance of beneficial intestinal microflora and decreased the abundance of harmful bacteria.Moreover,the abundance of Marvinbryantia was increased.Osteoking alleviated the decrease in metabolism of D-arginine and D-ornithine,sphingolipid,and galactose metabolism(P<0.05)and inhibited lysine degradation,the sulfur relay system,and propanoate metabolism(P<0.05).Conclusions Osteoking has hypoglycemic properties and improves the intestinal microflora imbalance in db/db mice.

Objective To investigate the role and mechanism of miRNAs in alcoholic liver injury in rats.Methods Thirty male SD rats were randomly divided into model and control groups.The model group was gavaged with 56%liquor and the control group was gavaged with distilled water for 8 weeks.Liver tissue was collected,miRNAs were analyzed,and target genes of differentially expressed miRNAs were predicted by a rat miRNA chip.Gene ontology(GO)and KEGG pathway enrichment analysis were used to understand the function of differentially expressed miRNA target genes.A differentially expressed miRNA-mRNA-pathway regulatory network was constructed using Cytoscape to further screen important regulatory miRNAs versus important pathways.RT-qPCR was performed for selected miRNAs to validate the expression analysis.Results Twelve differentially expressed miRNAs(P<0.05,Fold change≥2)were screened out,including two upregulated and 10 downregulated miRNAs by comparative analysis of microarray data between model and control groups.GO classification annotation of differential miRNA target genes showed close associations between differentially expressed miRNAs and biological functions such as signal transduction,metabolic processes,antioxidant activity,cell killing,enzyme regulatory activity and biological regulation.Differentially expressed miRNA target genes in KEGG pathway analysis revealed that the AMPK signaling pathway,PI3K-Akt signaling pathway,Hippo signaling pathway,Wnt signaling pathway,cancer,autophagy,insulin resistance,Ras signaling pathway,and other signaling pathways might play major regulatory roles in alcoholic liver injury lesions.Hub miRNAs and pathways screened by constructing the differentially expressed miRNA-mRNA-pathway regulatory network were miR-145-5p,miR-107-3p,miR-297,Hippo signaling pathway,cancer,PI3K-Akt signaling pathway,and AMPK signaling pathway.qRT-PCR validated the gene expression trends,and gene chip result were consistent.Conclusions We established an miRNA profile of alcoholic liver injury in rats,which suggests that miR-145-5p,miR-107-3p,and miR-297 play major roles in the process of alcoholic liver pathology.

Objective To study the therapeutic value of Mongolian drug hatagaqi-7 for wound healing of diabetic ulcers in rats and preliminarily explore its molecular mechanism in regulating hypoxia-inducible factor-1α(HIF-1α).Methods Adult male SD rats were randomly divided into control,diabetes,Mongolian drug,and cytokine groups.Except in the control group,the other three groups were treated with an intraperitoneal injection of streptozotocin to establish the diabetes model.Ulcer wounds were prepared in the mouse back of the four groups.One week later,the Mongolian drug group was treated with hatagaqi-7,and the cytokine group was treated with recombinant bovine basic fibroblast growth factor for 2 consecutive weeks.Fasting blood glucose(FBG),wound area,wound pathology,expression levels of advanced glycation end products(AGEs),receptor of AGE(RAGE),HIF-1α and vascular endothelial growth factor(VEGF),secreted levels of interleukin-1β(IL-1β),interferon-γ(IFN-γ),and malondialdehyde(MDA),and the total antioxidant capacity(T-AOC)were assessed.Results FBG of diabetes,Mongolian drug and cytokine groups was higher than that in the control group(P<0.05),and no significant difference was observed among the three groups(P>0.05).Compared with the control group,the ulcer wound area,scope of unrepaired tissue,expression levels of AGEs and RAGE,and secreted levels of IL-1β,IFN-γ,and MDA in wound tissue of the diabetes group were increased,and T-AOC and expression levels of HIF-1α and VEGF of the diabetes group were decreased(P<0.05).Compared with the diabetes group,the ulcer wound area,scope of unrepaired tissue,expression levels of AGEs and RAGE,and secreted levels of IL-1β,IFN-γ,and MDA in wound tissue of Mongolian drug and cytokine groups were decreased,T-AOC and expression levels of HIF-1α and VEGF in Mongolian drug and cytokine groups were increased(P<0.05),and indexes of the Mongolian drug group were better than those of the cytokine group.Conclusions Mongolian drug hatagaqi-7 promotes ulcer wound healing in diabetic rats,the inhibiton of AGE and RAGE expression and induction of HIF-1 α are the possible molecular mechanism.