Vascular calcification is one of the major complications of chronic kidney disease (CKD), which could be further accelerated by the osteogenic transition and apoptosis of smooth muscle cells, thereby ...advancing the progression of renal diseases and increasing the mortality rate of cardiovascular events. MicroRNA is a kind of key regulator in the phenotypic transition of vascular smooth muscle cells (VSMCs), but its role remains unclear in VSMCs. In this study, VSMCs were stimulated by platelet-derived growth factors - BB (PDGF-BB) in varying concentrations to establish the VSMC dysfunction models. The relative expression of miR-29a-5p was quantified via the quantitative real-time polymerase chain reaction (qRT-PCR). The proliferation of VSMCs was determined via the BrdU method, analysis of cell cycle via flow cytometry, and the migration of VSMCs via Transwell assay. Expression of γ-secretase activating protein (GSAP) and markers of VSMC differentiation, including α-SMA, SM-22α, SMMHC and Calponin, was quantified via the Western blot. The targeting relationship between the 3'-UTR of miR-29a-5p and GSAP was validated through the dual-luciferase reporter gene assay. As a result, we found that PDGF-BB could trigger a decrease of miR-29a-5p in a time- and dose-dependent manner (P < 0.05). Overexpression of miR-29a-5p could curb the effect of PDGF-BB on the proliferation and migration of VSMCs while upregulating the expression of markers of differentiation (P < 0.05). In addition, the expression of GSAP was also affected by the negative regulation of miR-29a-5p, while the restoration of GSAP eliminated the effect of miR-29a-5p on the VSMCs partially (P < 0.05). Moreover, vascular calcification models were also established in the CKD rats, suggesting that the inhibition of GSAP could prevent PTH-induced vascular calcification in CKD rats. In conclusion, miR-29a-5p could inhibit the PDGF-BB-induced proliferation, migration and phenotypic transition of VSMCs via targeting GSAP. Thus, miR-29a-5p/GSAP might be a potential target for the treatment of vascular calcification.
This study aimed to explore the role of miR-363-3p in renal fibrosis (RF) in vitro. HK-2 cells were treated with transforming growth factor (TGF)-β1 for 72 h to establish an in vitro model of RF. ...Subsequently, western blot analysis and reverse transcription-quantitative PCR were used to detect the protein and mRNA expression levels of RF markers in TGF-β1-treated HK-2 cells, respectively. The results showed that the protein and mRNA expression levels of TGF-β2, α-smooth muscle actin (SMA), fibronectin, vimentin, collagen II and N-cadherin were increased, while the protein and mRNA expression levels of E-cadherin were decreased in TGF-β1-treated HK-2 cells. The level of miR-363-3p was significantly decreased in TGF-β1-treated HK-2 cells. TargetScan indicated that TGF-β2 was a direct target gene for miR-363-3p, which was further verified using dual luciferase reporter gene assays. Further analyses revealed that the increased protein and mRNA expression levels of TGF-β2, α-SMA, fibronectin, vimentin, collagen II, N-cadherin, increased phosphorylated-Smad3 protein level, and decreased E-cadherin protein and mRNA expression in TGF-β1-treated HK-2 cells were significantly reversed by miR-363-3p mimics. However, all the effects were suppressed by a TGF-β2-plasmid. The results suggested that miR-363-3p plays a protective role in RF by regulating the TGF-β2/Smad3 signaling pathway.
It is acknowledged that Radix Rehmanniae Praeparata (RR) can regulate hormone metabolism, reduce blood glucose, resist aging, help to sedate patients and promote diuresis. The study aims to ...investigate the mechanism of how RR influences the development of renal fibrosis by regulating the miR-122-5p/PKM axis.
Unilateral ureteral obstruction (UUO) was applied to induce renal fibrosis in mice
, and human tubular epithelial HK2 cells treated by transforming growth factor-β (TGF-β1) were used to induce renal fibrosis
. Interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α) in mouse serum were detected by Enzyme-linked immunosorbent assay (ELISA); fibronectin (FN) and type I collagen (Col-I) in renal tissue were detected by Western blotting; serum creatinine (Cr) and blood urea nitrogen (BUN) were analyzed by kits. Hematoxylin-eosin (HE) staining and Masson staining were utilized to assess the degree of pathological damage and fibrosis. Cell viability and apoptosis in the
model were detected by MTT and Flow cytometry. Dual-luciferase reporter assay was performed to determine intermolecular targeting relationships.
RR could inhibit IL-6 and TNF-α levels, decrease the levels of FN and Col-I and improve the renal function indexes (serum Cr and BUN) in UUO mice (all P<0.05). In addition, RR was able to promote the up-regulation of miR-122-5p expression in UUO mice
(P<0.05). MiR-122-5p expression was down-regulated and PKM expression was up-regulated in HK2 cells treated with TGF-β1 (all P<0.05). RR inhibited renal fibrosis progression by regulating the miR-122-5p/PKM axis. Inhibition of miR-122-5p or overexpression of PKM could promote apoptosis of TGF-β1-treated HK2 cells, inhibit their viability, aggravate fibrosis, and attenuate the protective effect of RR on the cells. The protective effect of RR promoted by overexpression of miR-122-5p was partially counteracted by PKM.
RR can inhibit renal fibrosis progression by regulating the miR-122-5p/PKM axis.
Distributed learning is an effective tool to process big data. An easy and effective distributed learning approach is the divide and conquer method. It first partitions the whole data set into ...multiple subsets. A base learning algorithm is then applied to each subset. Finally the results from these subsets are coupled together. In the classification setting, many classification algorithms can be used in the second stage. Typical ones include the logistic regression and support vector machines. For the third stage, both voting and averaging can be used as the coupling strategies. In this thesis, empirical studies are done to thoroughly compare the effectiveness of these two coupling strategies. Averaging is found to be more effective in most scenarios.
The chromatographic fingerprint of Chinese traditional medicine, Yinhuang Oral Liquid together with its original materials Radix Scutellariae and Flos lonicera was established by high performance ...liquid chromatography (HPLC). At the same time, baicalin and chlorogenic acid in the Yinhuang Oral Liquid were also quantitatively determined. The analysis was performed on a Lichrospher C18 column (250 mm x 4.6 mm i. d. , 5 microm) and with gradient elution. The optimum conditions were explored. The mobile phase A was methanol and the mobile phase B was 0.1% H3PO4 aqueous solution. The wavelength of 254 nm was selected for detection. Finally, the data of fingerprints of 6 samples of Yinhuang Oral Liquid were processed with two different mathematic methods. This proposed method demonstrated good stability for the 6 samples of Yinhuang Oral Liquid. In the comparison of the common peaks in chromatogram of Yinhuang Oral Liquid with those of crude herbs, most of them could be matched well.
Cancer risk assessment of polycyclic aromatic hydrocarbons (PAH) is complicated by several of these compounds exerting a promoter action leading to high tumour incidences at high doses. Cancer risks ...at low doses corresponding to the uptake from air and food in the general environment would best be estimated on the basis of measurement of in vivo target doses of genotoxic (mutagenic) intermediates and a determination of mutation frequency per unit of dose. In experiments ultimately aiming at a risk assessment of environmental PAH from in vivo doses benzoapyrene (BaP) was chosen as a model. γ-Radiation has earlier been used as a reference standard in cancer risk estimation of genotoxic chemicals where dose equivalents (rad-equivalents) have been shown to give reliable risk estimates for several alkylating agents. Variation in dose of BaP diolepoxide between organs was studied by measurement of deoxyguanosine-N2 adducts in DNA after administration of BaP by gavage to mice of a strain with reduced DNA repair (Xpa–/–). The adduct levels in spleen, forestomach, stomach and small intestine were approximately the same; with the adduct level in spleen as reference it was twice as high in liver and lung and about half as high in colon tissue. A chemical or radiation dose is proportional to the cumulative frequency of putatively premutagenic changes (premutagenic hits) in DNA. The mutation frequency per premutagenic hit (genotoxic chemicals) and per unit of dose (γ-radiation) were calculated from acutely exposed V79 cells in order to determine the mutagenic effectiveness of each agent. Based on the mutagenic effectiveness determined in this study 10–4 Gy can be regarded equally effective in causing phenotypically expressed HPRT mutations as the dose of BaP which causes the formation of one deoxyguanosine-N2 adduct per cell.
With the spatial coherence of X-rays and high flux and brightness of the 3rd generation synchrotron radiation facility, X-ray phase contrast microscopy （XPCM） at Shanghai Synchrotron Radiation ...Facility （SSRF） can provide high resolution dynamic imaging of low electron density materials in principle. In this paper, we investigated the cavitation and water-refilling processes in rice and bamboo leaves utilizing XPCM at SSRF. The occurrence of xylem cavitation was recorded in vivo. The study also revealed that under different dehydration conditions, cavitation occurs in different degrees, and therefore, the refilling process is different. The results demonstrate that SSRF can provide high enough fluxes to study dynamic processes in plants in real-time, and XPCM is expected to be a promising method to reveal the mechanisms of cavitation and its repair in plants nondestructively.
Bats are connected with the increasing numbers of emerging and re-emerging viruses that may break the species barrier and spread into the human population. Coronaviruses are one of the most common ...viruses discovered in bats, which were considered as the natural source of recent human-susceptible coronaviruses, i.e. SARS-COV and MERS-CoV. Our previous study reported the discovery of a bat-derived putative cross-family recombinant coronavirus with a reovirus gene p10, named as Ro-BatCoV GCCDC1. In this report, through a two-year follow-up of a special bat population in one specific cave of south China, we illustrate that Ro-BatCoV GCCDC1 persistently circulates among bats. Notably, through the longitudinal observation, we identified the dynamic evolution of Ro-BatCoV GCCDC1 in bats represented by continuously recombination events. Our study provides the first glimpse of the virus evolution in one longitudinally observed bat population cohort and underlines the surveillance and pre-warning of potential interspecies transmittable viruses in bats.
Recent studies have shown the important role of proinflammatory cytokines and chemokines in the pathogenesis of atherosclerosis and diabetes mellitus (DM). Interferon-inducible protein of 10kD ...(IP-10/CXCL 10), a member of the C-X-C chemokine superfamily, is a potent chemoattractant for activated T lymphocytes and is reported to be involved in various disease states including atheroma plaque formation, inhibition of tumor angiogenesis and maintenance of podocyte function. However, the involvement of IP-10 in type 2 DM, especially in its vascular and renal complications, is largely unknown. To elucidate the etiopathological role of IP-10 in type 2 DM, we measured the concentrations of IP-10 together with IFN-γ, TNF-α, IL-18, IL-6 and MCP-1 in plasma samples from 103 type 2 DM patients with various degrees of nephropathy. A significant difference in the plasma level of IP-10 was observed between the patients and the control subjects (183.3±12.5pg/ml vs 65.6±9.3pg/ml, p<0.05). IP-10 correlated IL-18, IL-6, TNF-α and MCP-1. The IFN-γ level was below the detectable range. IP-10 levels became higher with the progression of nephropathy: IP-10 levels were 148.9±14.5, 174.2±17.2 and 231.9±31.3pg/ml in patients with an urinary albumin creatinine ratio of <30, 30 to 300 and >300μg/mg Cr, respectively. Similarly, IL-18, IL-6, MCP-1 and TNF-α levels in patients with overt albuminuria were significantly higher as compared with those without albuminuria (IL-18, 367.3±45.6 vs 203.5±17.6pg/ml; IL-6, 1.61±0.26 vs 0.87±0.13pg/ml; TNF-α, 1.83±0.48 vs 0.61±0.07pg/ml; p<0.05, respectively) in consistent with previous reports. These results suggested that IP-10 may have an etiopathogenic role in type 2 DM and diabetic nephropathy as one of the downstream effectors of proinflammatory cytokines.
PDF
