OBJECTIVES: To investigate the protective effect of Yiqi Yangyin Huazhuo Tongluo Formula (YYHT) against high glucose-induced injury in mouse renal podocytes (MPC5 cells) and the possible mechanism. METHODS: Adult Wistar rats were treated with 19, 38, and 76 g/kg YYHT or saline via gavage for 7 days to prepare YYHT-medicated or blank sera for treatment of MPC5 cells cultured in high glucose (30 mmol/L) prior to transfection with a miR-21a-5p inhibitor or a miR-21a-5p mimic. The changes in miR-21a-5p expressions and the mRNA levels of FoxO1, PINK1, and Parkin in the treated cells were detected with qRT-PCR, and the protein levels of nephrin, podocin, FoxO1, PINK1, and Parkin were detected with Western blotting. Autophagic activity in the cells were evaluated with MDC staining. The effect of miR-21a-5p mimic on FoxO1 transcription and the binding of miR-21a-5p to FoxO1 were examined with luciferase reporter gene assay and radioimmunoprecipitation assay. RESULTS: MPC5 cells exposed to high glucose showed significantly increased miR-21a-5p expression, lowered expressions of FoxO1, PINK1, and Parkin1 mRNAs, and reduced levels of FoxO1, PINK1, parkin, nephrin, and podocin proteins and autophagic activity. Treatment of the exposed cells with YYHT-medicated sera and miR-21a-5p inhibitor both significantly enhanced the protein expressions of nephrin and podocin, inhibited the expression of miR-21a-5p, increased the mRNA and protein expressions of FoxO1, PINK1 and Parkin, and upregulated autophagic activity of the cells. Transfection with miR-21a-5p mimic effectively inhibited the transcription of FoxO1 and promoted the binding of miR-21a-5p to FoxO1 in MPC5 cells, and these effects were obviously attenuated by treatment with YYHT-medicated sera. CONCLUSIONS YYHT-medicated sera alleviate high glucose-induced injury in MPC5 cells by regulating miR-21a-5p/FoxO1/PINK1-mediated mitochondrial autophagy.
Animals ; MicroRNAs/genetics* ; Podocytes/pathology* ; Drugs, Chinese Herbal/pharmacology* ; Autophagy/drug effects* ; Rats, Wistar ; Protein Kinases/metabolism* ; Rats ; Forkhead Box Protein O1 ; Mice ; Mitochondria/drug effects* ; Ubiquitin-Protein Ligases/metabolism* ; Glucose ; Diabetic Nephropathies ; Male ; Membrane Proteins/metabolism* ; Intracellular Signaling Peptides and Proteins

Objective:To analyze the related risk factors for vertebral artery tortuosity, and explore the mechanism of vertebral artery tortuosity.Methods:Two hundred and eighty-two patients accepted head/neck and MR angiography in our hospital from October 2016 to October 2017 were selected. The tortuosity degrees of vertebral artery were measured and calculated by PACS system. The differences of tortuosity degrees of vertebral arteries in different age groups were compared. Correlation analysis was performed to determine the correlation between vertebral artery tortuosity and both clinical data and and biochemical levels, and multivariate linear regression analysis was performed to determine the independent risk factors for vertebral artery tortuosity.Results:The tortuosity degrees of the left and right vertebral arteries in these patients ranged from 5.1% to 72.6%. The tortuosity degrees of vertebral arteries in patients aged 40-49 years were significantly higher than those in patients aged 20-29 years and 30-39 years ( P<0.05). Correlation analysis showed that the tortuosity degree of the right vertebral artery was positively correlated with age and triglyceride level ( r=0.232, P=0.000; r=0.172, P=0.004); the tortuosity degree of the left vertebral artery was positively correlated with triglyceride level ( r=0.123, P=0.043). Multivariate regression analysis showed that age ( 95%CI: 0.059-0.194, P=0.000) and triglyceride level ( 95%CI: 0.173-1.942, P=0.019) were independent risk factors for right vertebral artery tortuosity. Triglyceride level ( 95%CI: 0.041-2.559, P=0.043) was independent risk factor for left vertebral artery tortuosity. Conclusion:There are congenital developmental factors associated with vertebral artery tortuosity; some nurture factor, as triglyceride level, may promote its development.

Objective To explore the effect of astragalus extracts on adherence, migration, cell viability, microvascular formation, and eNOS expression in bone marrow-derived endothelial progenitor cells (EPCs) of rats. Methods EPCs were cultured, isolated and identified in vitro and divided into four groups: low titer group (10-4 g/L of astragalus extracts), middle titer group (10-3 g/L of astragalus extracts), high titer group (10-2 g/L of astragalus extracts) and control group. The effects of astragalus extract on adhesion, migration or microvascular formation were observed under the inverted microscope and com?pared between all groups;Cell viability was detected by MTT assay;mRNA transcription and protein expression levels of en?dothelial nitric oxide synthase (eNOS) in EPCs were detected by reverse transcription PCR (RT-PCR) and Western blot re?spectively. Results Compared with the control group, cell adhesion, migration and microvascular formation of EPCs all en?hanced with addition of astragalus extracts in a dose dependent manner (F=15.256, 13.633, 97.549 respectively, and P <0.05 in all cases); Cell vitality of EPCs increased with administration of astragalus extracts in a time and dose dependant manner. (F=9.755 for time and F=10.18 for dose). mRNA transcription and protein expression of eNOS in EPCs were up-reg?ulated with addition of astragalus extracts in a dose dependent manner (F=56.356 and 77.125 respectively, and P<0.05 in both cases). Conclusion Astragalus extracts play important role in angiogenesis in EPCs probably through up-regulating expressions of eNOS.

AIM:Todeterminetheeffectofsalviaextractonangiogenesisofthemyocardiumintheratswith myocardial infarction (MI) and to analyze its possible mechanism .METHODS: Left coronary artery of Sprague-Dawley rats was ligated to establish a MI model .The rats were randomly divided into MI model group , 3 different dose groups of salvia (10, 20 and 40 mg? kg-1? d-1), and sham operation group.Each group consisted of 8 rats.The rats in all treat-ment groups were orally administered with the salvia extract , and the rats in MI group and sham operation group were fed with the same volume of saline .The rats were sacrificed 4 weeks later .The hemodynamic changes of the rats were deter-mined , and the segmental heart samples were used for morphological observation by hematoxylin and eosin staining , Masson staining, or electron microscopic analysis.The expression of vascular endothelial growth factor (VEGF) and cluster of dif-ferentiation 34 ( CD34 ) was analyzed according to immunohistochemistry .RESULTS: Compared with sham operation group, the morphological changes of the myocardium in MI group were disordered , part of myocardial cell outline disap-peared , and obvious fibrosis in the necrosis myocardial tissue and fuzzy or disappearing microvascular ultrastructure were al -so observed .Compared with MI group , the number of new microvessels in all the treatment groups increased obviously , and the morphological changes of the endothelial cells were relatively complete according to electron microscopy .Compared with sham operation group , the protein expression of VEGF and CD 34 in the cytoplasm of the myocardial tissues in MI group in-creased only a little .Compared with MI group , the protein expression of VEGF and CD 34 in the cytoplasm of the myocardi-al tissues in all treatment groups increased significantly ( P<0.01 ) .CONCLUSION: Salvia extract obviously promotes angiogenesis of the myocardial tissues in the rats after myocardial infarction .

Aim To explore the effect of protein kinase D1 (PKD1 )on adherence,migration,proliferation, microvascular formation,and eNOS expression in bone marrow-derived endothelial progenitor cells(EPCs)of rats.Method The EPCs were isolated from bone marrow of rats,cultured and detected,the effects of PKD1 and its specific blocking agent CID755673 on adhesion,migration,proliferation,or microvascular formation were observed,as well as mRNA and protein expression of endothelial nitric oxide synthase (eNOS ) in EPCs.Results PKD1 significantly promoted adhe-sion,migration,proliferation and microvascular forma-tion of EPCs,and upregulated the mRNA and protein expression of eNOS in EPCs,according to the cell cul-ture experiments of EPCs in vitro.Conclusion PKD1 has the role of angiogenesis through regulation of EPCs,which might be dependent on eNOS.

BACKGROUND:Results of recent studies demonstrated the modulation of thymosin β4 on hair cycle and regeneration, but the mechanism of action remains unclear.
OBJECTIVE:To investigate the mechanism by which thymosinβ4 increases hair regeneration through Wnt signal pathway.
METHODS:After the mouse model of depilation was established using rosin/paraffin mixed agents, the experimental animals were randomly assorted to three different groups, including low-dose, high-dose and control groups, and a dose of 0.3μg/50μL, 3μg/50μL thymosinβ4 and PBS was administered on the depilated backs every 12 hours, respectively. Then photography, hematoxylin-eosin staining, immunohistochemistry and in situ hybridization were applied to observe the growth of hair, and the expressions ofβ-catenin and LEF-1 mRNA in different groups at different time were quantitatively evaluated.
RESULTS AND CONCLUSION:The hair growth of the low-dose group was faster than that of the other groups. Hematoxylin-eosin staining demonstrated inflammatory cel s infiltration in the dermis after depilation, and the number of hair fol icles that were in the phase of anagen was much more than the other groups as time went by. Immunohistochemistry ofβ-catenin showed the accumulation of intra-cel ularβ-catenin in the low-dose group at the bulge of fol icles assessed by integrated absorbance analysis (P<0.05), so did the in situ hybridization of LEF-1 mRNA. Low-dose thymosinβ4 accelerates hair growth through Wnt signal pathway by elevating the level ofβ-catenin and LEF-1 mRNA.