• Volume 28,Issue 6,2020 Table of Contents
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    • Effects of compound Qishao Jiangya tablet on inflammatory factors, AP-1 and MCP-1 in vascular endothelial cells stimulated with TNF-α and co-cultured with smooth muscle cells

      2020, 28(6):733-741. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 001

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      Abstract: Objective To observe the expression levels of inflammatory factors, AP-1, and MCP-1 in TNF-α- stimulated vascular endothelial cells and co-cultured with smooth muscle cells following treatment with the compound Qishao Jiangya tablet, and explore the mechanism by which the compound Qishao Jiangya tablet inhibits vascular remodeling. Methods A co-culture system was established using vascular endothelial cells and smooth muscle cells. Stimulation with TNF-α was used to mimic the hypertensive inflammatory environment. The cultured cells were randomly divided into seven groups: normal control, model, blank serum control, MRS2578, irbesartan tablet drug-containing serum, compound Qishao Jiangya tablet-containing serum, and MRS2578 + compound Qishao Jiangya tablet-containing serum. Except the normal control group, all groups were stimulated with 10 ng / mL TNF-α; the irbesartan tablet drug-containing serum, compound Qishao Jiangya tablet-containing serum, and MRS2578+compound Qishao Jiangya tablet-containing serum were treated with 10% drug-containing serum; the MRS2578 and MRS2578+compound Qishao Jiangya tablet drug-containing serum groups were treated with 10 μm P2Y6 receptor blocker MRS2578. After 24 hours of modeling and drug intervention, CCK8 assays were used to detect the survival rates of endothelial cells and smooth muscle cells. ELISA was used to detect the level of IL-8 in the culture supernatant. Immunofluorescence, Western Blot, and quantitative PCR were used to detect endothelial expression levels of AP-1 and MCP-1. Results Compared with the normal control group, the survival rates of smooth muscle and endothelial cells decreased significantly in the model and blank serum control groups, while the levels of IL-8 were significantly higher (P < 0. 01). Compared with the normal control group,AP-1 and MCP-1 in endothelial cells in the model and blank serum control groups were detected by fluorescence intensity value of immunofluorescence, gray value ratio by Western Blot, and mRNA levels by quantitative PCR were significantly increased (P< 0. 01). Compared with the model group, the survival rates of smooth muscle and endothelial cells were significantly increased in the compound Qishao Jiangya tablet-containing serum groups, while the levels of IL-8 were significantly lower (P < 0. 05). AP-1 and MCP-1 in endothelial cells were detected by fluorescence intensity value of immunofluorescence, gray value ratio by Western blotting, and mRNA levels by quantitative PCR in the compound Qishao Jiangya tablet-containing serum groups were significantly reduced compared with the model group(P < 0. 01). Conclusions Following stimulation with TNF-α, the compound Qishao Jiangya tablet can improve the survival rates of endothelial cells and smooth muscle cells, reduce the levels of AP-1 and MCP-1, prevent endothelial cell inflammation, improve endothelial damage, and protect normal function of smooth muscle cells.

    • Effects of risedronate sodium combined with simvastatin on dexamethasone-induced bone loss in rats

      2020, 28(6):742-748. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 002

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      Abstract: Objective To analyze the effects of simvastatin and risedronate sodium, either alone or in combination, on bone loss induced by dexamethasone in rats. Methods Twelve-week-old female Sprague-Dawley rats were divided into five groups ( n = 8 ) as follows: control group, osteoporosis model group subcutaneously injected with dexamethasone (2. 5 mg, twice / week), simvastatin group (5 mg / ( kg·d)), risedronate group (0. 1 mg / ( kg·d)), and combination group. Eight weeks later, all rats were sacrificed, and the left femurs were harvested for bone mineral density (BMD) and three-point bending testing. In addition, micro-CT was performed on the left tibia. The right femurs were embedded in paraffin for immunohistochemical analysis of OPG and RANKL expression, and the right tibia samples were subjected to RNA extraction to assess OPG and RANKL expression. Results BMD was lowest in the osteoporosis group (0. 207 ± 0. 02) g / cmP>2P>> , and BMD was markedly lower in the simvastatin ( 0. 226 ± 0. 023) g / cmP>2P>> and risedronate (0. 237 ± 0. 021) g / cmP>2P>> groups than in the control (0. 282 ± 0. 013) g / cmP>2P>> and combination groups (0. 257 ± 0. 012) g / cmP>2P>> . The maximum load in the osteoporosis group was significantly lower than that in the control and combination groups. BV/ TV and Tb.N were significantly lower in the osteoporosis group than in the other groups, whereas the opposite trend was noted for Tp.Sp. BV/ TV was significantly lower in the simvastatin (21. 6 ± 2. 8) and risedronate groups (21. 9 ± 2. 6) than in the control (29. 5 ± 2. 7) and combination groups (25. 3 ± 2. 9). Tb.Sp was significantly higher in the simvastatin and risedronate groups than in the control group. On immunohistochemistry, OPG expression was significantly lower in the osteoporosis, risedronate, and combination groups than in the control group, whereas its expression was significantly higher in the simvastatin and combination groups than in the osteoporosis group. RANKL expression was significantly higher in the osteoporosis, simvastatin, risedronate, and combination groups than in the control group and significantly lower in the combination group than in the osteoporosis group. Meanwhile, OPG expression was significantly lower in the osteoporosis group (0. 74 ± 0. 17) than in the control (1. 00 ± 0. 16) and simvastatin groups (1. 27 ± 0. 19). RANKL expression was significantly higher in the osteoporosis group than in the other groups, and its expression was significantly higher in the simvastatin group (1. 31 ± 0. 16) than in the control group (1. 00 ± 0. 18). Conclusions Dexamethasone induced bone loss in rats, and this effect was more obviously prevented by combined treatment with simvastatin and risedronate than by either drug alone.

    • Effect of miR-328 in the exercise-induced improvement of the L-type calcium channel expression in thoracic aorta from hypertensive rats

      2020, 28(6):749-758. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 003

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      Abstract: Objective The present study investigated the effect of microRNA-328 (miR-328) during the aerobic exercise-induced improvement of voltage-gated L-type Ca2+ channel ( LTCC) expression in thoracic aorta smooth muscle cells from spontaneously hypertensive rats ( SHRs). Methods Twelve-week-old male normotensive Wistar Kyoto rats (WKYs) and SHRs randomly were separated into a control group (SHR-C and WKY-C) and an exercise group (SHR-EX and WKY-EX). Rats in exercise were subjected to moderate-intensity treadmill training for 12 weeks. The cardiovascular responses of rats were then detected by femoral arterial and venous cannulation. The thoracic aorta was stained with hematoxylin and eosin ( HE) for histological assays. The mRNA expression levels of LTCC α1C and β1 subunits and the expression of miR-328 were detected by qPCR. The protein expression levels of LTCC α1C and β1 subunits were detected by Western Blot. For in vitro analysis, primary vascular smooth muscle cells (VSMCs) were isolated and cultured from the thoracic aorta of WKYs. Both miR-328 mimic and miR-328 inhibitor were transfected into cultured arterial myocytes to produce miR-328 overexpression or silencing. The mRNA and protein expression levels of LTCC α 1C and β1 subunits were detected. Results Aerobic exercise significantly reduced body weight, blood pressure, and heart rate in SHRs. Aerobic exercise significantly reduced the wall thickness of thoracic aorta in SHRs. Exercise significantly reduced the pressor response to norepinephrine and the depressor response to nifedipine ( LTCC blocker) in SHRs. The mRNA and protein expression levels of LTCC α1C and β1 subunits were significantly upregulated in the SHR-C group, compared with the WKY- C group, while the mRNA and protein expression levels of LTCC α1C and β1 subunits were downregulated in the SHR-EX group, compared with the SHR-C group. miR-328 expression was significantly lower in the SHR-C group than that in the WKY-C group; this expression significantly increased after aerobic exercise. In the transfection experiment, compared with the negative control ( NC) group, the mRNA and protein expression levels of α1C and β1 subunits were significantly downregulated in the miR-328 mimic transfected group; these expression was significantly upregulated in the miR-328 inhibitor transfected group. Conclusions Regular aerobic exercise may effectively reduce blood pressure in SHR and enable miR-328 to inhibit LTCC α1C and β1 subunit expression. This mechanism might contribute to exercise-mediated improvement of LTCC channel remodeling in hypertensive thoracic aorta.

    • Establishment and identification of an immune-humanized mouse model of prostate cancer

      2020, 28(6):759-764. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 004

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      Abstract: Objective An ideal animal model for the study of prostate cancer (Pca) immunotherapy is needed. We aimed to establish a mouse model humanized for the immune system and then transplant these mice with human Pca cells. Methods To humanize mice with a reconstructed human immune system, peripheral blood mononuclear cells (PBMCs) were separated from fresh human peripheral blood by density gradient centrifugation and injected into NOD-scid Il2rg- / - mice via the caudal vein. Peripheral blood from these mice was collected at the 3rd, 4th, 5th and 6th weeks following injection, and dynamic changes in human CD45+CD3+ T cells were monitored using flow cytometry analysis. At the 3rd week after humanization, cells from the Pca cell line 22Rv1 were subcutaneously implanted into the right flank of humanized mice. Tumor growth was monitored twice a week. When the tumor grew to about 1000 mmP>3P> , the mice were euthanized. The infiltration of human immune cells in the peripheral blood, spleen and tumor tissues of humanized mice was analyzed by flow cytometry, immunohistochemistry (IHC) and hematoxylin and eosin (HE) staining. Results High levels of CD45+CD3+ T cells were detected in the peripheral blood of mice from 3 to 6 weeks after PBMC transplantation. The mice were euthanized at the 6th week and the result of HE and IHC staining demonstrated that the spleen and tumor tissues of the mice were infiltrated by human immune cells CD4+ and CD8+ T cells. Conclusions The immune-humanized mouse model of Pca was successfully established. This model showed high levels of human immune T cells in the peripheral blood, spleen and Pca tissues, laying a foundation for the construction of an ideal preclinical model of immunotherapy for Pca.

    • Detection of MRTF-A-binding gene expression in the MCAO / R mouse model via RIP-Seq

      2020, 28(6):765-772. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 005

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      Abstract: Objective To detect the expression of RNA-bound myocardin-related transcription factor A (MRTF- A)-binding genes using RIP-Seq technology in a mouse middle cerebral artery occlusion (MCAO) / reperfusion model and explore the potential mechanism of action of MRTF-A. Methods C57BL/ 6 mice were randomly divided into the sham and cerebral ischemia / reperfusion (I/ R) injury groups. The model of focal MCAO was constructed using the suture method. After 24 h of reperfusion, total brain tissue protein was extracted. Furthermore, the expression profile of MRTF-A-binding genes was detected using immunoprecipitation plus high-throughput sequencing. Additionally, the differentially expressed genes were analyzed using GO and KEGG. Results Compared with the findings in the sham group, 429 genes were differentially expressed (203 upregulated and 226 downregulated genes) in the cerebral I/ R group. GO molecular function analysis revealed that the differentially expressed genes were mainly enriched in RNA binding and Poly(A) RNA binding. KEGG pathway analysis illustrated that 10 pathways were significantly enriched, among which the estrogen signaling pathway was most enriched. Conclusions The expression profile of MRTF-A-binding genes was significantly altered by cerebral I/ R injury, which provides a theoretical basis for in-depth exploration of the molecular mechanism of MRTF-A.

    • Comparison of mouse models of influenza virus infection established with two respiratory tract infection modes

      2020, 28(6):773-778. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 006

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      Abstract: Objective To explore the differences in modeling influenza virus infection with two different respiratory tract infection modes in mice; and provide a reference for choosing the appropriate infection model for pathogenic research on influenza and developing vaccines and drugs. Methods The A/ Puerto Rico / 8 / 34 (H1N1) virus strain was selected to infect C57BL/ 6 mice by intranasal instillation and aerosol inhalation. The mice were weighed daily and their clinical symptoms were observed visually. Mice were killed on the 3rd, 7th, or 14th day after infection, and the lungs were weighed and used for virus assay, pathological observation and cytokine detection. Results Both infection modes successfully established an influenza virus infection model in a mouse, and their general trends of disease progression were similar. However, there are some differences in infection characteristics between the two models. Compared with the intranasal instillation group, weight loss occurred earlier in the aerosol inhalation group, and the lung index and viral load increased significantly on the 3rd day after infection (P< 0. 05). The lesion range and degree of inflammatory cytokine infiltration were also significantly increased in the aerosol group. Furthermore, the levels of cytokines interleukin( IL) -1α and IL-6 in the lung increased significantly on the 3rd and 7th days after infection, and tumor necrosis factor-α increased significantly on the 7th day (P< 0. 05). Conclusions Both modes of infection established a model of influenza in the mouse,but aerosol inhalation caused more obvious inflammatory infiltration and cytokine expression in the lung at an early stage of infection. The aerosol inhalation mode led to a more ideal animal model of influenza virus infection.

    • Generation of human HBB mutant knock-in porcine fibroblasts using CRISPR / Cas9-mediated homologous recombination

      2020, 28(6):779-787. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 007

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      Abstract: Objective The high-frequence mutant genotype Condons41 / 42 (-CTTT) of human thalassaemia- related HBB gene ( hHBB) was knocked into genome of the porcine fibroblasts by using CRISPR-Cas9 Site-specific recombination systems. Then the transfected fibroblasts were screened to establish homozygous cell lines with positive genotype. Methods (1) Synthesize the human HBB mutant with Condons41 / 42 (-CTTT) and cloned the left and right homology arm located at the flanking sequence of the pig HBB gene. (2) Then the HBB mutant was inserted between the left and right homology arm using infusion PCR method to building a final homologous recombination vector. ( 3) The sgRNA targeting pig HBB gene were designed using online software and then were verified its activity in pig PK15 cells. (4) Bama pig fetal fibroblasts were co-transfected with CRISPR/ Cas9 vector, sgRNA and pGH-LA-hHBB-RA using by electroporation and then were screened to establish single clone. Results (1) We synthesized full length of the hHBB CDS with a mutant Condons41 / 42 (-CTTT). The left and right homology arm of 1. 062 kb and 1. 024 kb were cloned from Bama pig genome. Then we constructed a final vector pGH-LA-hHBB-RA. ( 2) Two sgRNAs targeted pig HBB gene were validated in PK15 cells and used for further experiments. (3) Finally, we identified 15 single-cell clones of the porcine fibroblast carry the human HBB gene mutant. Conclusions We generated porcine fibroblast carrying the human HBB gene mutant using CRISPR/ Cas9. Our study provides key materials for scientist to generate pig models of human thalassemia.

    • Effect of the Astragalus dose in Buyang Huanwu decoction on erectile dysfunction in rats with cavernous nerve injury

      2020, 28(6):788-795. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 008

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      Abstract: Objective To explore the therapeutic effects of two different doses of Buyang Huanwu decoction containing Astragalus on erectile dysfunction (ED) in rats with cavernous nerve injury. Methods Thirty-two Sprague- Dawley ( SD) rats were randomly divided into four groups: sham operation model, 30 g ( model + Buyang Huanwu decoction containing 30 g Astragalus) and 120 g (model + Buyang Huanwu decoction containing 120 g Astragalus). The cavernous nerve of the rats was surgically clamped (but not cut) to establish the rat model of ED caused by cavernous nerve injury. Five days after the operation, a 30 g or 120 g dose of Astragalus was used for the gavage intervention of Buyang Huanwu decoction, or the same volume of normal saline ( sham and model groups) was given, once a day for 30 consecutive days. After 30 days, the intracavernous pressure ( ICP) of the rat corpus cavernosum was measured. Masson staining was used to observe the content of smooth muscle and collagen deposition in the spongy tissue. Fluorescence and western blotting were used to detect changes in neuronal nitric oxide synthase ( nNOS), neurofilament light polypeptide (NF-L), growth associated protein 43 (GAP-43) expression in each group. Results Compared with the sham group, the model group showed significantly decreased ICP ( P < 0. 01), decreased area of penile smooth muscle / collagen ( P < 0. 01), and significantly decreased levels of NF-L, nNOS and GAP43 (P< 0. 01). Compared with the model group, the rats in the 30 g and 120 g Astragalus groups showed significantly increased ICP (P < 0. 01), increased area of penile smooth muscle / collagen (P< 0. 01), and significantly increased levels of nNOS, NF-L and GAP43 (P< 0. 01). Compared with the 30 g group, the rats in the 120 g Astragalus group showed significantly increased ICP (P< 0. 05), increased area of penile smooth muscle / collagen (P< 0. 05), and significantly increased levels of nNOS, NF-L and GAP43 (P< 0. 01). Conclusions Buyang Huanwu decoction containing Astragalus obviously improved ED in cavernous nerve injury model rats. The higher dose of Astragalus (120 g), which is used in the original prescription of “Medical Forest Correction”, was more efficacious.

    • Effects of human umbilical cord mesenchymal stem cells on autophagy in the naturally aging rat hippocampus

      2020, 28(6):796-804. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 009

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      Abstract: Objective To explore whether transplantation of human umbilical cord mesenchymal stem cells (hUCMSCs) can improve learning, memory and cognitive decline by regulating autophagy levels in naturally aging rats. Methods Sprague-Dawley rats were reared in a barrier environment to the age of 24 months. Behavioral studies select aging rats with obvious cognitive decline; the rats were divided into a cell transplantation group ( group H) and a normal saline group (group C). For comparison, a normal group ( group N) of 3-month-old rats was included. Rats in group H were injected with 2×10P>6P>> hUCMSCs in a volume of 500 μL through the tail vein. Rats in groups C and N were injected with an equal volume of sterile saline through the tail vein. Injections were performed once per week for 4 weeks. Changes in learning and memory ability were assessed using the Morris water maze, Y maze, and novel object recognition test. Changes in rat hippocampal neuron morphology were assessed using hematoxylin-eosin staining. Relative expression levels of hippocampal LC3II/ I, Beclin1, and p62 were assessed using Western Blot. Results Behavioral analyses showed that the learning and memory abilities were significantly better in groups H and N than in group C. HE staining of nerve cells in the CA1 and DG areas of the hippocampus revealed more regular arrangements, more complete cell structure, and more intact nuclei in groups H and N. Conversely, the number of neurons in the CA1 area was reduced, cell arrangement was loose, nuclei were shrunken, and degenerative disease was present in group C. p62 protein content was significantly lower in groups H and N than in group C, while Beclin1 and LC3II/ I protein levels were significantly higher in groups H and N than in group C. Conclusions Intravenous injection of hUCMSCs improved learning, memory, and cognitive decline in naturally aging rats, presumably by restoration of autophagy.

    • Continuous hemodiafiltration combined with enalapril improves renal and intestinal function in rats with severe acute pancreatitis

      2020, 28(6):805-811. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 010

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      Abstract: Objective To explore the beneficial effects of continuous hemodiafiltration (CHDF) combined with enalapril on renal and intestinal function in rats with severe acute pancreatitis ( SAP). Methods Sixty Sprague-Dawley rats were randomly divided into a control group and four treatment groups: SAP model, CHDF, enalapril, and CHDF + enalapril. Rats in the treatment groups were induced with SAP using sodium taurocholate. After modeling, the CHDF group was given CHDF for 12 h, and the enalapril group was given enalapril 10 mg / kg / d by gavage, for 1 day. The CHDF + enalapril group was given both treatments simultaneously. Histopathological changes in the pancreas, kidney and intestinal mucosa were observed by hematoxylin and eosin staining. The plasma levels of endotoxin (ET) and D-lactic acid (D-LC), and serum levels of amylase (AMY), blood urea nitrogen ( BUN) and creatinine ( Cr) were measured. A fluorescein isothiocyanate-dextran permeability test was used to examine intestinal permeability. Western blotting was used to detect expression of α-vascular smooth muscle actin (α-SMA), E-cadherin, high mobility groupbox protein 1 ( HMGB1) and occludin. Results Compared with the model, CHDF + enalapril had the following effects in rats with SAP: effectively alleviated damage to the pancreas, kidney and intestinal mucosal tissues; significantly reduced intestinal mucosal permeability, the levels of serum AMY, BUN and Cr and plasma ET and D-LC, and expression of α-SMA and HMGB1 in tissues; and significantly increased expression of E-cadherin and occludin. The combined action effect was better than that of either single treatment. Conclusions CHDF combined with enalapril significantly improved renal and intestinal tissue damage caused by SAP, which contributed to the recovery of renal and intestinal function.

    • Effects of dietary calcium and vitamin D supplementation on osteoclast activity and bone mass in a rat ankle fracture model

      2020, 28(6):812-817. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 011

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      Abstract: Objective To investigate the effects of calcium and vitamin D ( Ca / VitD) supplementation on osteoclast activity and bone mass after hand and foot trauma. Methods Eight-week-old male mice were randomly divided into three groups: control ( C), Ca / VitD deficiency ( D), and Ca / VitD supplementary diet groups ( S). Group S underwent hand and foot trauma surgery after 8 weeks of standard feeding. Mouse serum analysis, micro-computed tomography (CT), histoeconometric analysis, immunohistochemical analysis, and gene expression analysis of fracture calli were used to investigate whether Ca / VitD deficiency could impair bone repair and cause bone loss after injury. The study also investigated whether Ca / VitD supplementation in the diet from the time of fracture could enhance fracture healing. Results Compared with the findings in group C, BMD and bone mass were significantly decreased in group D, whereas the fibrous tissue volume was increased. In the fracture callus, the number and surface of osteoclasts in group D were significantly enhanced. Compared with the result in group D, the bone mass in the callus was significantly increased in group S, whereas the amount of fibrous tissue was significantly decreased after Ca / VitD supplementation. In addition, group S exhibited a higher fracture healing rate. In group S, expression of the C-terminal telopeptide of type I collagen was decreased, whereas that of alkaline phosphatase and X-linked phosphate regulatory gene was increased. Meanwhile, the serum levels of iFGF23 and cFGF23 were significantly higher in group S than in groups D and C. However, the iFGF23: cFGF23 ratio was not different among the groups. Conclusions Ca / VitD supplementation after hand and foot trauma reduced osteoclast activity, increased bone mass, and suppressed bone absorption in mice with Ca / VitD deficiency. These findings should have guiding significance for clinical postoperative nursing.

    • Effects of Oct4 and Cdx2 on differentiation of bovine embryonic inner cell mass and trophectoderm

      2020, 28(6):818-823. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 012

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      Abstract: Objective To investigate the effects of Oct4 and Cdx2 knockdown on the expression of genes involved in ICM (Inner Cell Mass) and TE (Trophoectoderm) differentiation in cattle, and to elucidate the roles of Oct4 and Cdx2 in the early development of bovine embryos. Methods Oct4 and Cdx2 genes were knocked down in bovine embryos by RNA interference. Real-time quantitative PCR and immunofluorescence staining were used to observe the effects of knockdown on early embryo development and gene expression. Results Specific siRNAs against Oct4 and Cdx2 inhibited Oct4 and Cdx2 expression, respectively, in bovine embryos. Oct4 knockdown had no effect on embryonic cleavage and eight-cell proportions, although the blastocyst proportion was significantly reduced; moreover, the mRNA levels of Cdx2, Sox2, and Nanog were significantly reduced (P < 0. 05). Cdx2 knockdown inhibited the expression of Oct4 mRNA (P < 0. 05). Conclusions Oct4 controls the differentiation of bovine embryos by regulating the mRNA expression levels of Cdx2, Sox2, and Nanog.

    • Protective effect and associated mechanism of action of estrogen on oxidative stress injury of MC3T3-E1 osteoblasts in mice

      2020, 28(6):824-830. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 013

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      Abstract: Objective To explore the protective effect and associated mechanism of action of estrogen on oxidative stress injury of mouse MC3T3-E1 osteoblasts (OB). Methods MC3T3-E1 cells cultured in vitro were divided into a blank control group, H2O2 group (300 μmol / L), H2O2+ 0. 1 μmol / L estrogen group, H2O2+ 1 μmol / L estrogen group, H2O2+ 10 μmol / L estrogen group, and H2O2+ 1 mmol / L NAC group, which were incubated with the corresponding concentrations of the drugs. The proliferative activities of cells in each group were determined by CCK-8. The apoptosis rate and mitochondrial membrane potential (MMP) were determined by flow cytometry. The levels of malondialdehyde (MDA) and superoxide dismutase (SOD) were determined using kits. The level of reactive oxygen species (ROS) was determined by a fluorescent probe method . The expression of Smad5, Runx2, Bax, Bcl-2, and cleaved Caspase-3 proteins was determined by western blotting. Results Compared with those in the blank control group, the proliferative activities of cells were significantly decreased (P< 0. 05), apoptosis rate and positive rate of JC-1 cells were significantly increased (P< 0. 05), levels of MDA and ROS were significantly increased (P< 0. 05), SOD activity was significantly decreased (P< 0. 05), expression of Smad5, Runx2, and Bcl-2 proteins was significantly decreased ( P < 0. 05), and expression of Bax and cleaved Caspase-3 proteins was significantly increased in the H2O2 group (P< 0. 05). Compared with those in the H2O2 group, proliferative activities of cells were significantly increased (P< 0. 05), apoptosis rate and positive rate of JC-1 cells were significantly decreased (P< 0. 05), levels of MDA and ROS were significantly decreased (P< 0. 05), SOD activity was significantly increased (P< 0. 05), expression of Smad5, Runx2, and Bcl-2 proteins was significantly increased (P< 0. 05), and expression of Bax and cleaved Caspase-3 proteins was significantly decreased in the H2O2+ 1 μmol / L estrogen group, H2O2+ 10 μmol / L estrogen group, and H2O2 + 1 mmol / L NAC group (P< 0. 05). Conclusions Estrogen may reduce the level of ROS in MC3T3-E1 cells damaged by oxidative stress and improve the differentiation ability of cells by activating the expression of the Smad5 / Runx2 signal axis.

    • Thinking of study on animal model of TCM syndrome of congestive heart failure

      2020, 28(6):831-836. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 014

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      Abstract:In this review, we summarized research approaches involved in animal model preparation of modern medicine, TCM syndrome, and the combination of disease and syndrome. Based on it, we proposed to set up integration animal model including congestive heart failure and TCM syndromes, which is realistic and strong operability, and will be of great advantage to carry out the experimental study, objectification and standardization research of TCM syndromes of congestive heart failure.

    • Research progress of the mouse model of HBV-related hepatocellular carcinoma

      2020, 28(6):837-844. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 015

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      Abstract:Viral infections cause approximately 12% of cancers worldwide and approximately 1. 3 million cancer- related deaths each year. However, representative in vivo models for all types of tumor viruses are lacking. Liver cancer is the fourth leading cause of cancer-related death in the world, and hepatocellular carcinoma (HCC) accounts for 80% of all liver cancer cases. Hepatitis B virus (HBV)-associated HCC is particularly a public health issue in China, which consists of more than 60% of domestic patients. Chronic HBV infection is the critical etiological factor for liver cancer. However, curable treatments for these patients are still lacking. The prerequisite for developing effective cancer drugs for HBV- associated HCC is to disrupt the mechanism of HBV interaction with the host. The mouse model of HBV-induced liver cancer has been remarkably advanced in recent decades, and multiple novel models have been developed by precision medicine. This review comprehensively summarizes the viral and oncogenic properties of in vivo models of HBV-induced HCC, with a focus on genetically engineered mouse and humanized mouse models. In addition, we discuss the benefits and caveats of each model and present a selection of the most important findings obtained from the respective systems.

    • Research progress and evaluation of animal models of hepatorenal fibrosis

      2020, 28(6):845-852. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 016

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      Abstract:Fibrosis of the liver and kidneys is a pathological process in which extracellular matrix ( ECM) is overdeposited and parenchyma cells are reduced by continuous damage to liver and kidney cells. Liver and kidney fibrosis is an inevitable process of chronic liver and kidney diseases. Except for organ transplantation, the early stage of fibrosis is now considered the key part of treatment for chronic disease. There are currently many rodent models of liver or kidney fibrosis, but only a few models with combined liver and renal fibrosis are available for research and drug screening. This paper summarizes the modeling method and mechanisms of common liver and kidney fibrosis, and composite models. We also compare the merits and demerits of different method to provide a reference for the future establishment and improvement of more clinically relevant compound animal models of liver and kidney fibrosis.

    • Advances in the application of micro-CT in animal models of vascular calcification

      2020, 28(6):853-856. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 017

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      Abstract:Vascular calcification is an ectopic calcification phenomenon caused by excessive deposition of calcium salts in the vascular wall. This pathological manifestation is common in diseases such as atherosclerosis, diabetes, hypertension, and chronic renal insufficiency; it is an important risk factor for cardiovascular and cerebrovascular diseases. Current computed tomography ( CT ) techniques can completely reconstruct calcified arteries and provide accurate quantitative information. High resolution micro-CT can distinguish and quantify areas of macroscopic and microscopic vascular calcification. CT combined with 18F-sodium Flfluoride ( 18F-NaF) micro-PET/ CT can enhance the detection of microcalcification in vivo and aid in understanding the roles of signaling pathways and drugs in the development of vascular calcification. Furthermore, three-dimensional micro-CT can be combined with histopathological, immunohistochemical, and proteomic analysis method to provide complementary information regarding a single vascular segment. Advances in micro-CT techniques may provide additional method for the analysis of vascular calcification in animals in vivo.

    • Research progress in a mouse model of endometriosis

      2020, 28(6):857-863. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 018

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      Abstract:Endometriosis (EM) is defined by the presence of active endometrial tissue in the lining of the uterine cavity. Because its etiology and pathogenesis are unclear, the diagnosis of EM and the combined use of Traditional Chinese medicine and Western medicine are difficult, and thus, EM has become a research hotspot in recent years. Animal experiments are important for exploring the etiology, pathogenesis, and drug therapy of EM. This paper discussed the selection of animals, construction method , and advantages and disadvantages of various EM mouse models in an effort to provide a reference for model selection in animal research of EM.

    • Research progress on lipopolysaccharide-induced pneumonia animal models

      2020, 28(6):864-869. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 019

      Abstract (1380) HTML (0) PDF 768.41 K (3855) Comment (0) Favorites

      Abstract:Pneumonia involves the inflammation of terminal airways, alveoli and pulmonary interstitium, and can be caused by pathogenic microorganisms, immune damage, physical and chemical factors, allergies and other conditions. In recent years, many domestic and foreign scholars have used lipopolysaccharide to induce the incidence of pneumonia in animal models. However, there are many differences in these models, including the induction dose, modeling method , and selection of evaluation and detection indicators. Therefore, we sought to review the literature on animal models of lipopolysaccharide-induced pneumonia to provide a reference for the study of the pathogenesis, prevention and treatment of pneumonia.

    • Research progress in the construction of type II diabetes animal models

      2020, 28(6):870-876. DOI: 10. 3969 / j.issn.1005-4847. 2020. 06. 020

      Abstract (2118) HTML (0) PDF 802.63 K (4403) Comment (0) Favorites

      Abstract:Diabetes mellitus is a metabolic disease characterized by hyperglycemia and caused by many factors. Diabetes poses a serious threat to human health. It is of great significance to construct a corresponding animal model of diabetes to study its pathogenesis, prevention, and diagnosis and to screen new drugs. On the basis of summarizing the construction of animal models of spontaneous and induced type II diabetes, this paper focused on the construction of animal models of type II diabetes using genetic engineering technology and discussed the advantages and disadvantages of various construction method to provide appropriate animal models for unraveling the pathogenesis and treatment of diabetes.

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