The aim of this experiment was to investigate the effects of daidzein(DZ)on mammary gland development in mice during pregnancy.A total of 36 pregnant mice were randomly divided into a control group(treated with sterile PBS),a DZ group(DZ working fluid by intraperitoneal injection)and a E2 group(E2 working fluid by intraperitoneal injection),once daily,for 7 days.Af-ter normal delivery of pregnant mice,6 mice were selected from each group on the 1st and 10th day of the lactation period,then executed after blood sampling,and all mammary gland tissue were taken for the following experiments:1)mammary gland tissue was weighted and the mammary gland index was calculated;preparation of mammary gland tissue sections,HE staining;2)ELISA of estradiol,prolactin and insulin-like growth factor concentrations in serum;3)Western blot and RT-qPCR were used to analyze the expression of cell cyclin proteins and ER-a,ER-β proteins and genes in mammary gland tissues.The results showed that:compared with control group,DZ signif-icantly increased the area ratio of acinar in the mammary gland of mice(P＜0.05),decreased the area ratio of adipose tissue and increased proliferative expansion of adenohypophysis;DZ highly significantly increased serum PRL levels(P＜0.01)and serum IGF-1 content,reduced serum E2 levels;DZ significantly up-regulated the expression of CyclinD1,CyclinD3,PCNA and ER-β pro-teins(P＜0.05),and the mRNA level of ER-β gene was significantly up-regulate(P＜0.01).In conclusion,DZ can promote cell proliferation and division by increasing the expression of cell cycle proteins and proliferation proteins,promotes the proliferation of mammary gland follicles and in-creases mammary gland weight during pregnancy in mice,and at the same time increase the serum levels of IGF-1 and PRL in the female rats,which jointly promote the development of the mamma-ry glands during lactation.

The aim of this experiment was to investigate the effects of daidzein(DZ)on mammary gland development in mice during pregnancy.A total of 36 pregnant mice were randomly divided into a control group(treated with sterile PBS),a DZ group(DZ working fluid by intraperitoneal injection)and a E2 group(E2 working fluid by intraperitoneal injection),once daily,for 7 days.Af-ter normal delivery of pregnant mice,6 mice were selected from each group on the 1st and 10th day of the lactation period,then executed after blood sampling,and all mammary gland tissue were taken for the following experiments:1)mammary gland tissue was weighted and the mammary gland index was calculated;preparation of mammary gland tissue sections,HE staining;2)ELISA of estradiol,prolactin and insulin-like growth factor concentrations in serum;3)Western blot and RT-qPCR were used to analyze the expression of cell cyclin proteins and ER-a,ER-β proteins and genes in mammary gland tissues.The results showed that:compared with control group,DZ signif-icantly increased the area ratio of acinar in the mammary gland of mice(P＜0.05),decreased the area ratio of adipose tissue and increased proliferative expansion of adenohypophysis;DZ highly significantly increased serum PRL levels(P＜0.01)and serum IGF-1 content,reduced serum E2 levels;DZ significantly up-regulated the expression of CyclinD1,CyclinD3,PCNA and ER-β pro-teins(P＜0.05),and the mRNA level of ER-β gene was significantly up-regulate(P＜0.01).In conclusion,DZ can promote cell proliferation and division by increasing the expression of cell cycle proteins and proliferation proteins,promotes the proliferation of mammary gland follicles and in-creases mammary gland weight during pregnancy in mice,and at the same time increase the serum levels of IGF-1 and PRL in the female rats,which jointly promote the development of the mamma-ry glands during lactation.

BACKGROUND:Irisin,a myokine isolated from the transmembrane protein FNDC5 by muscle cells during exercise,has the function of inducing the browning of white adipose tissue,but its effect on lipotoxicity-induced osteogenic differentiation and the mechanism is unclear. OBJECTIVE:To investigate the effect of irisin on the osteogenic ability of palmitic acid-induced bone marrow mesenchymal stem cells and the mechanism of action. METHODS:CCK-8 assay was used to detect the effect of different concentrations of palmitic acid on the proliferation of mouse bone marrow mesenchymal stem cells and the effect of irisin on the proliferation of mouse bone marrow mesenchymal stem cells in the presence of palmitic acid.After pretreatment with irisin and palmitic acid for 24 hours,osteogenic differentiation of mouse bone marrow mesenchymal stem cells was induced by alkaline phosphatase staining as well as qRT-PCR was performed to detect the expression of osteogenesis-related genes on day 7 of osteogenic induction culture.The expression of proteins related to the AMPK/BMP2/SMAD signaling pathway was detected by western blot assay.Alizarin red staining was conducted on day 21 to detect osteogenic differences. RESULTS AND CONCLUSION:(1)The CCK-8 assay results suggested that the amplification of bone marrow mesenchymal stem cells was inversely proportional to the concentration of palmitic acid,but at 0.02 mmol/L concentration,palmitic acid had no significant effect on the amplification of bone marrow mesenchymal stem cells,and irisin did not affect the proliferation of bone marrow mesenchymal stem cells when its mass concentration was in the range of 0.1-20 μg/L.(2)Alkaline phosphatase staining and alizarin red staining showed that palmitic acid inhibited the osteogenic differentiation ability of bone marrow mesenchymal stem cells.Irisin improved palmitic acid-induced osteogenic inhibition of bone marrow mesenchymal stem cells.qRT-PCR results showed that palmitic acid could cause the downregulation of osteogenic-related genes,and irisin could inhibit this trend.(3)Western blot assay results showed that compared with the palmitic acid intervention group,irisin treatment enhanced AMPK/BMP2/SMAD signal transduction in bone marrow mesenchymal stem cells.It is found that irisin can improve the osteogenic differentiation ability of bone marrow mesenchymal stem cells pretreated with palmitic acid,and proposed that the specific mechanism might be mediated by AMPK/BMP/SMAD signaling pathway.

The goal of this study was to investigate the regulatory effect of angiotensin converting enzyme 2 (ACE2) on cellular inflammation caused by avian infectious bronchitis virus (IBV) and the underlying mechanism of such effect. Vero and DF-1 cells were used as test target to be exposed to recombinant IBV virus (IBV-3ab-Luc). Four different groups were tested: the control group, the infection group[IBV-3ab-Luc, MOI (multiplicity of infection)=1], the ACE2 overexpression group[IBV-3ab Luc+pcDNA3.1(+)-ACE2], and the ACE2-depleted group (IBV-3ab-Luc+siRNA-ACE2). After the cells in the infection group started to show cytopathic indicators, the overall protein and RNA in cell of each group were extracted. real-time quantitative polymerase chain reaction (RT-qPCR) was used to determine the mRNA expression level of the IBV nucleoprotein (IBV-N), glycoprotein 130 (gp130) and cellular interleukin-6 (IL-6). Enzyme linked immunosorbent assay (ELISA) was used to determine the level of IL-6 in cell supernatant. Western blotting was performed to determine the level of ACE2 phosphorylation of janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3). We found that ACE2 was successfully overexpressed and depleted in both Vero and DF-1 cells. Secondly, cytopathic indicators were observed in infected Vero cells including rounding, detaching, clumping, and formation of syncytia. These indicators were alleviated in ACE2 overexpression group but exacerbated when ACE2 was depleted. Thirdly, in the infection group, capering with the control group, the expression level of IBV-N, gp130, IL-6 mRNA and increased significantly (P < 0.05), the IL-6 level was significant or extremely significant elevated in cell supernatant (P < 0.05 or P < 0.01); the expression of ACE2 decreased significantly (P < 0.05); protein phosphorylation level of JAK2 and STAT3 increased significantly (P < 0.05). Fourthly, comparing with the infected group, the level of IBV-N mRNA expression in the ACE2 overexpression group had no notable change (P > 0.05), but the expression of gp130 mRNA, IL-6 level and expression of mRNA were elevated (P < 0.05) and the protein phosphorylation level of JAK2 and STAT3 decreased significantly (P < 0.05). In the ACE2-depleted group, there was no notable change in IBV-N (P > 0.05), but the IL-6 level and expression of mRNA increased significantly (P < 0.05) and the phosphorylation level of JAK2 and STAT3 protein decreased slightly (P > 0.05). The results demonstrated for the first time that ACE2 did not affect the replication of IBV in DF-1 cell, but it did contribute to the prevention of the activation of the IL-6/JAK2/STAT3 signaling pathway, resulting in an alleviation of IBV-induced cellular inflammation in Vero and DF-1 cells.
Animals ; Chlorocebus aethiops ; Humans ; Interleukin-6/genetics* ; Janus Kinase 2/pharmacology* ; Infectious bronchitis virus/metabolism* ; STAT3 Transcription Factor/metabolism* ; Angiotensin-Converting Enzyme 2/pharmacology* ; Cytokine Receptor gp130/metabolism* ; Vero Cells ; Signal Transduction ; Inflammation ; RNA, Messenger

Background and Objectives: Manipulating different signaling pathways via small molecules could efficiently inducecardiomyocytes from human induced pluripotent stem cells (hiPSC). However, the effect of transcription factors on the hiPSC-directed cardiomyocytes differentiation remains unclear. Transcription factor, p53 has been demonstrated indispensable for the early embryonic development and mesendodermal differentiation of embryonic stem cells (ESC).We tested the hypothesis that p53 promotes cardiomyocytes differentiation from human hiPSC. Methods: and Results: Using the well-characterized GiWi protocol that cardiomyocytes are generated from hiPSC via temporal modulation of Wnt signaling pathway by small molecules, we demonstrated that forced expression of p53 in hiPSC remarkably improved the differentiation efficiency of cardiomyocytes from hiPSC, whereas knockdown endogenous p53 decreased the yield of cardiomyocytes. This p53-mediated increased cardiomyocyte differentiation was mediated through WNT3, as evidenced by that overexpression of p53 upregulated the expression of WNT3, and knockdown of p53 decreased the WNT3 expression. Mechanistic analysis showed that the increased cardiomyocyte differentiation partially depended on the amplified mesendodermal specification resulted from p53-mediated activation of WNT3-mediated Wnt signaling. Consistently, endogenous WNT3 knockdown significantly ameliorated mesendodermal specification and subsequent cardiomyocyte differentiation. Conclusions These results provide a novel insight into the potential effect of p53 on the development and differentiation of cardiomyocyte during embryogenesis.

This article aimed at exploring the effects and protective mechanism of β-CM7 on renin angiotensin system (RAS) in diabetic rats myocardial tissue. We divided 32 male SD rats into 4 groups: control group, diabetic model control group, insulin (3.7x10(-8) mol/d) treatment group and β-CM7 (7.5x10(-8) mol/d) treatment group. After 30 days, all rats were decapitated and myocardical tissues were collected immediately. After injection, β-CM7 could decrease the content of Ang II, increase the content of Angl-7. And β-CM7 could improve the mRNA of AT1 receptor and Mas receptor. β-CM7 also could improve the mRNA of ACE and ACE2, enhance the activity of ACE and ACE2. These data confirmed tli β-CM7 could activate ACE2-Angl-7-Mas axis, negative passage in RAS, to inhibit the expression ACE mnRiJA and protein in rat myocardium, alleviate the myocardial tissue damage induced by Ang II. The effect of β-CM7 on inhibiting myocardium damage might be related to ACE/ACE2 passageway.
Angiotensin II ; metabolism ; Animals ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetic Cardiomyopathies ; drug therapy ; Endorphins ; pharmacology ; Male ; Myocardium ; metabolism ; pathology ; Peptide Fragments ; pharmacology ; Peptidyl-Dipeptidase A ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Receptor, Angiotensin, Type 1 ; metabolism ; Receptors, G-Protein-Coupled ; metabolism ; Renin-Angiotensin System

New epidemic broke out in recent year which was suspected to be caused by variant Muscovy duck parvovirus (MDPV). For this reason, new MDPV detection methods are needed for the new virus strains. In this study, a pair of primers were designed according to the full-length genome of MDPV strain SAAS-SHNH, which were identified in 2012, and were used to amplify the vp3 gene of MDPV by polymerase chain reaction. After being sequenced, the vp3 gene was subcloned into the prokaryotic expression vector PET28a. The recombinant plasmid was transformed into E. coli BL21 and induced with IPTG. SDS-PAGE and Western blotting analysis showed the MDPV vp3 gene was successfully expressed. After being purified by Ni2+ affinity chromatography system, the recombinant protein was used as antigen to immunize rabbits to obtain antiserum. Western blotting analysis showed that the acquired antiserum could react specifically with VP3 protein of J3D6 strain and MDPV vaccine strain. The antiserum could also be used for detection of cultured MDPV from primary duck embryo fibroblasts by immune fluorescence assay (IFA). It could be concluded that the VP3 protein and its antibody prepared in the research could be used for detection of VP3 antiserum and antigen respectively.
Animals ; Blotting, Western ; DNA Primers ; Ducks ; virology ; Electrophoresis, Polyacrylamide Gel ; Immune Sera ; biosynthesis ; Parvovirus ; Polymerase Chain Reaction ; Rabbits ; Recombinant Proteins ; genetics ; Viral Proteins ; genetics

The composition of S95-100 field mobile medical system was introduced, and some practices were summarized such as rational grouping, enhanced organization, proper allocation and etc. The problems of the system were analyzed from three aspects of the fixation of the internal devices, the rapid deployment in the plateau and maintenance in field conditions, and some measures were put forward to improve the medical support ability of the system in the plateau.

Objective To discuss the possibility of interventional diagnosis and treatment of uterine artery bleeding after curettage. Methods Select the emergent patients with uterine artery bleeding after curettage as the basis for study. Three women with uterine artery bleeding underwent femoral artery puncture and transcatheter uterine artery embolization. Bilateral selective artery angiographies were performed, and then underwent artery embolization after exhibiting the bleeding sites. Results Three patients were promptly and correctly diagnosed and arterial embolization were then attempted under DSA. No recurrence of bleeding during the angiographic and clinical follow up simultaneously with no serious complication. Conclusions Uterine arterial DSA and interventional embolization of uterine artery are effective in the diagnosing and treatting emergent uterine artery bleeding after curettage.

This paper reported the acute toxicity test and the long-term toxicity test of Tongbi Tablets in rats and dogs. The LD_(50) of mice was 5.29g/kg. For dogs following oral administration (0.513g/kg/day, which was a tenth part of mice'LD_(50)) for 6 months running, the toxic symptom of strychnine-like and an increase in BUN could be-observed after 4-month administration. Both recoveried after breaking off the administration. For 0.051g/kg/day dosage group of dogs(administered for 6 months running)and all dosage groups of rats, no toxic symptom was observed, suggesting that the clinical dosage of this medicine was safe.