Abstract: ObjectiveTo 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: Objective　To investigate the modeling process and model establishment rate of the N-methyl-Nnitrosourea(MNU)-induced lower jiao dampness-heat syndrome uremia bladder cancer (BC) model. Methods 　Batch samples were collected by bladder perfusion (MNU). HE staining and pathological observation of pathological tissues were carried out at six time points before, during, and after modeling to understand the formation and development of bladder carcinoma in situ. Results 　The experimental process was smooth, and no obvious urinary tract bleeding in rats occurred during the operation. Using this modeling technique, considerable bladder tumors were formed after week 8, and obvious epithelial hyperplasia, damage and large-scale tumor formation were observed in the bladder of rats in the model group. Conclusions　MNU induces an SD rat bladder cancer model with obvious model establishment and low mortality. The experimental data provide a reference to establish, improve and apply the bladder cancer model.

Abstract: Objective　To explore the effect of Dingkun pill on the PI3K/ AKT/ mTOR signaling pathway in rats with endometriosis (EM). Methods 　EM rat models were established by heterotopic transplantation of endometrial tissue and randomly divided into five groups: model group (M group), Dingkun pill low (1. 13 g/ kg) dose group (DKP-L group), Dingkun pill medium (2. 26 g/ kg) dose group (DKP-M group), Dingkun pill high (4. 52 g/ kg) dose group (DKP-H group) and gestrinone (60 mg/ kg) group (GES group) with 12 rats in each group. The abdomen was opened without transplantation of ectopic endometrial tissue in another 12 normal SD rats as sham operation group (Sham group). The rats were sacrificed after drug treatments, The mass and volume of ectopic endometrium were measured. The microvessel density (CD31-positive rate) and expression of VEGF and MMP-9 in rat ectopic endometrial tissue were assessed by immunohistochemical staining. Rat serum VEGF, MMP-9, iNOS and TNF-α levels were measured by enzymelinked immunosorbent assays. Expression of PI3K/ AKT/ mTOR pathway-related proteins in rat ectopic endometrial tissue was detected by Western blot. Results 　Compared with those in the sham group, the microvessel density, VEGF and MMP-9 expression, serum VEGF, MMP-9, iNOS and TNF-α levels, p-PI3K/ PI3K, p-AKT/ AKT and p-mTOR/ mTOR in ectopic endometrial tissue were significantly increased in the M group (P<0. 05). Compared with the findings in the M group, the ectopic endometrial volume, weight of the ectopic lesion, microvessel density of the ectopic endometrial tissue, VEGF and MMP-9 expression, serum VEGF, MMP-9, iNOS and TNF-α levels, p-PI3K/ PI3K, p-AKT/ AKT, and pmTOR/ mTOR in ectopic endometrial tissue were all decreased in the DKP-L, DKP-M, DKP-H groups, and dosedependent effects were observed in Dingkun pill low dose, Dingkun pill medium dose, and Dingkun pill high dose groups(P<0. 05). Compared with DKP-H group and GES group, no significant difference was found in the indicators (P>0. 05). Conclusions　Dingkun Dan reduces inflammation and inhibits ectopic endometrial growth in EM rats, which may be achieved by blocking the PI3K/ AKT/ mTOR signal pathway.

Abstract: Objective　To investigate the effects of CeO₂NP subacute exposure on body weight, organ structures, and redox indicators in male mice. Methods 　Thirty-five male ICR mice were randomly divided into five groups with seven mice in each group. Normal control and CeO₂NP (100, 500, 1000 and 5000 μg/ kg)groups were included. Mice were injected with CeO₂NPs intraperitoneally every other day for 28 days. Results 　Compared with the control group, no significant difference was found in weight gain (P>0. 05), but significant differences were observed in the liver coefficient, epididymal coefficient, and sperm survival rate (P<0. 01). In the 5000 μg/ kg group, CeO₂NPs were deposited in the liver and spleen, and granuloma was found SOD and GSH-Px activities in serum of the 100 μg/ kg group were increased, while MDA content was decreased, which indicated the antioxidant effect of CeO₂NPs. With the increase in dose, oxidative stress induced by CeO₂NPs was enhanced. Conclusions　Low-dose-CeO₂NPs are safe and exert an antioxidant effect. With the increase in dose, the toxicity of CeO₂NPs also increases gradually.

Abstract: Objective　To explore the extraction and purification method of Kupffer and hepatic stellate cells from mouse liver and provide references and suggestions for the separation and extraction method ology of primary nonparenchymal cells from mouse liver. Methods 　After in vivo collagenase perfusion digestion, various reagents and method, such as Percoll and OptiPrep, were used to extract C57BL/6 mice Kupffer and hepatic stellate cells, and evaluate their purity by flow cytometry and immunofluorescence. Results 　The two-layer Percoll method to extract Kupffer cells and the two-layer OptiPrep method to extract hepatic stellate cells were feasible, and purity reached >90%. The cell yield was 1~2×10⁷ / liver, and the cell survival rate was >90%. After 48 hours of primary cell culture, the number of F4/80-positive Kupffer cells and α-SMA-positive hepatic stellate cells reached > 90%. Conclusions 　The separation and extraction method of Kupffer and hepatic stellate cells from mouse liver are perfect, reliable, cost-effective, and reproducible.

Abstract:Vascular dementia (VD) is caused by cerebrovascular diseases, either hemorrhage or ischemic damage in the brain, with ischemia being the most common. In recent years, increasing efforts have been made to study the etiology, pathogenesis, and prevention of VD. The establishment of appropriate animal models to study the mechanism of VD and explore the efficacy of VD treatments has become an important issue in this research field. On the basis of conventional method, such as bilateral occlusion of common carotid arteries (2VO) and four-vessel occlusion, researchers have modified these method to improve stability with better reflection of the clinical manifestations of VD. This review summarizes these modified method and discusses possible cellular and molecular mechanisms and their advantages and disadvantages.

Abstract:Neurodegenerative diseases (NDs) are closely related to the central nervous system and characterized by morphological abnormalities and progressive loss of function in specific neuron groups. The main NDs include Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, multiple sclerosis and Huntington’ s disease. However, no direct therapies for NDs exist. In recent years, single cell RNA-sequencing (scRNA-seq) has been widely used in various NDs. The pathogenesis of NDs is closely related to morphology of immune cells, and the pathogenesis mainly involves mitochondrial function, glucose metabolism, inflammation, and synaptic transmission. Induced pluripotent stem cells are a potential therapy for NDs. Ultimately, we review the application of scRNA-seq to various NDs and provide a reference for prevention and treatment of NDs.

Abstract:The establishment of effective animal models is crucial for disease research. Dietary induction is a common method to establish animal models of non-alcoholic fatty liver disease (NAFLD). The common animal models of diet-induced NAFLD mainly include high fat and sugar, high fat and cholesterol, and choline-deficient diet-induced models. Because of the various nutrients ingested in different modeling method, pathological characteristics of the liver, such as fatty deformation, inflammation, and fibrosis, are different. Additionally, animal models vary in terms of disease progression, disease severity, and applicable studies. This study analyzed and compared the common animal models of NAFLD induced by various diets in terms of modeling method, modeling time, pathological characteristics, applicable research, and related advantages and disadvantages to provide a reference for NAFLD researchers to select animal models.

Abstract:Systemic sclerosis (SSc) is an autoimmune disease characterized by vascular lesions and skin fibrosis, which causes structural and functional damage to multiple organs. The pathogenesis of SSc is complex and may be related to genetic and environmental factors as well as abnormal immune mechanisms. Autophagy is an evolutionarily conserved process of self-degradation in the body and participates in the pathogenesis of autoimmune diseases through various pathways. This article reviews the research progress on the role of autophagy in the angiopathy and fibrosis of SSc by regulating immune cells and mediating signaling pathways.

Abstract:The liver is the most common organ for tumor spread, and expression of miRNA is crucial for liver metastasis. In this study, research progress of related miRNAs in regulating liver metastases from malignant tumors in the digestive system was collated and analyzed. By searching related literature, this article provides an introduction to the role of miRNAs in liver metastasis of colorectal, gastric, pancreatic, and gallbladder cancers, which helps with the diagnosis, treatment, and research of tumor liver metastasis.

Abstract:Organ transplantation is the main treatment for organ failure. Functional protection of donor organs during ex vivo transportation is critical for the success of organ transplantation. How to protect the functions of donor organs during in vitro transportation is an important issue in the field of organ transplantation research. In a hypoxic environment, transcriptional activity of a series of genes in cells is activated. These genes are mainly involved in angiogenesis, iron metabolism, glucose metabolism, and cell proliferation/ survival. In aerobic organisms, hypoxia-inducible factor-1α (HIF-1α) is involved in the regulation of the expression of various genes to maintain homeostasis of tissues and cells under hypoxic conditions, thereby adapting to the hypoxia. Many studies have shown that the HIF-1α pathway plays an important role in protecting isolated organs from cold ischemic injury during cold storage. HIF-1α has been a hot topic in research on the protective mechanism of cold ischemic injury of isolated organs. Regulating the HIF-1α-related signaling pathway is expected to be a new strategy to maintain organ functions during cold storage of isolated organs.

Abstract:The rapid development of the Chinese economy and the continuous advancement of urbanization have made the problem of air pollution more prominent, which may have an impact on people’s health. In recent years, a large number of studies have emerged in the academic community both domestically and internationally. Many cohort studies have shown that during pregnancy, the surrounding air pollutants are relatively high, which may increase the risk of adverse pregnancy outcomes. So now people are paying more attention to the impact of air pollutants on the health of pregnant women or fetuses. This article provides a brief review of the impact of atmospheric pollutants on some adverse pregnancy outcomes and their possible biological mechanisms.

Abstract:Kidneys filter bloodand regulate fluid and electrolyte balances. However, the kidney is susceptible to toxicity of various compounds, Resulting in renal damage. Hence, prevention and treatment of renal injury is a hot research topic. Zebrafish is an ideal animal model, because it is closely related to humans in terms of morphology, physiology, and kidney functions and its response to compounds. In this article, we review the method and evaluation indexes of zebrafish kidney injury modeling.