Search

The present work aimed to investigate the activities and underlying mechanisms of dioscin against alcoholic liver fibrosis (ALF). In vivo liver fibrosis in mice was induced by an alcoholic liquid diet, and in vitro studies were performed on activated HSC-T6 and LX2 cells treated with lipopolysaccharide. Our results showed that dioscin significantly attenuated hepatic stellate cells (HSCs) activation, improved collagen accumulation, and attenuated inflammation through down-regulating the levels of myeloid differentiation factor 88 (MyD88), nuclear factor κB (NF-κB), interleukin (IL)-1, IL-6 and tumour necrosis factor-α by decreasing Toll-like receptor (TLR)4 expression both in vivo and in vitro. TLR4 overexpression was also decreased by dioscin, leading to the markedly down-regulated levels of MyD88, NF-κB, transforming growth factor-β1 (TGF-β1), α-smooth muscle actin (α-SMA) and type I collagen (COL1A1) in cultured HSCs. Suppression of cellular MyD88 by ST2825 or abrogation of NF-κB by pyrrolidine dithiocarbamate eliminated the inhibitory effects of dioscin on the levels of TGF-β1, α-SMA and COL1A1. In a word, dioscin exhibited potent effects against ALF via altering TLR4/MyD88/NF-κB signaling pathway, which provided novel insights into the mechanisms of this compound as an antifibrogenic candidate for the treatment of ALF in the future.

The mechanisms of the natural product dioscin against non-alcoholic fatty liver disease (NAFLD) are unclear. Thus, the purpose of the present study was to further confirm its effects of prevention and then to elucidate the potential mechanisms underlying its activity in mice. High-fat diet (HFD)-induced C57BL/6J mice and ob/ob mice were used as the experimental models. Serum and hepatic biochemical parameters were determined, and the mRNA and protein expression levels were detected. The results indicated that dioscin alleviated body weight and liver lipid accumulation symptoms, increased oxygen consumption and energy expenditure, and improved the levels of serum and hepatic biochemical parameters. Further investigations revealed that dioscin significantly attenuated oxidative damage, suppressed inflammation, inhibited triglyceride and cholesterol synthesis, promoted fatty acid β-oxidation, down-regulated MAPK phosphorylation levels, and induced autophagy to alleviate fatty liver conditions. Dioscin prevents diet induced obesity and NAFLD by increasing energy expenditure. This agent should be developed as a new candidate for obesity and NAFLD prevention.

To investigate the clinical efficacy of pre-operative simulation using a three-dimensional (3D) printing model for surgical treatment of old and complex tibial plateau fractures. Forty-two patients with old and complex tibial plateau fractures were retrospectively reviewed from January 2014 to January 2018, which were divided into a conventional planning group (n = 22) and a planning with 3D printing group (n = 20). In the planning with 3D printing group, preoperative equal-ratio fracture models prepared using the 3D printing technique were used to perform pre-operative simulation and guide the real surgical operation. In the conventional planning group, the operation was performed based on pre-operative computed tomography (CT) images. Surgery duration, blood loss and the number of fluoroscopy during operations were recorded. During follow-up, the quality of fracture reduction and complications were also recorded. Knee functions were evaluated using the hospital for special surgery (HSS) scoring system. The operation time, blood loss and the number of fluoroscopy during operation in the planning with 3D printing group were less than that in the conventional planning group (P P = 0.05). The complication rate was 15% in the planning with 3D printing group and 31.82% in the conventional planning group. At the final follow-up evaluation, the mean HSS score was 86.05 ± 7.67 in the planning with 3D printing group and 79.09 ± 6.75 in the conventional planning group (P = 0.003). The rate of excellent results in the planning with 3D printing group was 70% and in the conventional planning group was 45.45% (P = 0.083). In conclusion, pre-operative simulation using a 3D printing model may be helpful for the treatment of old and complex tibial plateau fractures, which may be conducive to the pre-operative planning and to making the surgical procedure accurate and personalized. However, its clinical effectiveness need to be further assessed by a prospective randomized-controlled study.

To study the fractal dimensions of the soil particle size distributions (PSDs) within different plantations (of Pinus densiflora, Quercus acutissima, Robinia pseudoacacia, and Larix kaempferi) and evaluate PSDs as an indicator of the likelihood of Robinia pseudoacacia invasion, the soil porosity of 0–20 cm soil layers was measured at different plantations in the Yaoxiang Forest Farm, Shandong Province, China. The results showed that the fractal dimension (Dm) values varied from 2.59 to 2.70 among the different plantations and were significantly negatively correlated to sand content and positively correlated to silt content and clay content. Significant negative correlations were observed between Dm and both soil organic matter (SOM) (P P D1) and entropy dimension/capacity dimension (D1/D0) parameters were not significantly correlated with SOM, although significant correlations were found between SOM and each of D0, Δα, and Δf(α). Compared with the other plantations, the Robinia pseudoacacia plantation had higher nutrient contents, higher D0 and D1 values and lower Dm values. Based on principal component analysis (PCA) ordination, we concluded that Robinia pseudoacacia and Pinus densiflora shared a similar habitat and that Robinia pseudoacacia is more likely to invade Pinus densiflora plantations for soil.

Many long non-coding RNAs (lncRNAs) have been identified in several types of human pituitary adenomas and normal anterior pituitary, some of which are involved in the pathogenesis of pituitary adenomas. However, a systematic analysis of lncRNAs expressed at different developmental stages of normal pituitary, particularly in rats, has not been performed. Therefore, we contrasted two cDNA libraries of immature (D15) and mature (D120) anterior pituitary in rat that were sequenced on an Illumina HiSeq Xten platform, and a total of 29,568,806,352 clean reads were identified. Notably, 7039 lncRNA transcripts corresponded to 4442 lncRNA genes, and 1181 lncRNA transcripts were significantly differentially expressed in D15 and D120. In addition, 6839 protein-coding genes (

To assess anatomical changes in eyes with progressive myopia, we morphometrically examined the eyes of guinea pigs with lens-induced axial elongation. Starting at an age of 3–4 weeks, guinea pigs in the experimental group (n = 20 animals) developed unilateral lens-induced axial elongation by wearing goggles for 5 weeks compared to a control group of 20 animals without intervention (axial length:8.91 ± 0.08 mm versus 8.74 ± 0.07 mm; P P = 0.41), equator (P = 0.41), midpoint between equator and posterior pole (MBEPP) (P = 0.13) or posterior pole (P = 0.89). Retinal pigment epithelium (RPE) cell density decreased with longer axial length at the MBEPP (P = 0.04; regression coefficient beta = −0.33) and posterior pole (P = 0.01; beta = −0.40). Additionally, the thickness of the retina and sclera decreased with longer axial length at the MBEPP (P = 0.01; beta = −0.42 and P P P P

The cytotoxicities of three aconitum alkaloids- aconitine, hypaconitine and mesaconitine, and their abilities to bind DNA have been explored. Rat myocardial cells H9c2 were treated with aconitum alkaloids and assessed the cytotoxicities by using MTT assay and flow cytometry. Apoptosis was evidenced by the results of the annexin V/propidium iodide (PI) assay. Aconitine was found to be the most toxic in rat myocardial cells H9c2 in three aconitum alkaloids. At the same time, DNA adducts were isolated and then analyzed by UV-Vis spectroscopy after exposure to alkaloids, which indicated that three alkaloids could bind to DNA in rat myocardial cells H9c2. Furthermore, their binding modes were investigated by UV-Visible, fluorescence, DNA melting studies and ionic strength effect. Results indicated that the interaction between three alkaloids and DNA were intercalation coupled with electrostatic effect. The estimated binding constants were between 4.83 × 10

A simple, accurate, sensitive and robust assay that can rapidly and specifically measure the death of target cells would have applications in many areas of biomedicine and particularly for the development of novel cellular- and immune-therapeutics. In this study, we describe a novel cytotoxicity assay, termed the Matador assay, which takes advantage of the extreme brightness, stability and glow-like characteristics of recently discovered novel marine luciferases and their engineered derivatives. The assay involves expression of a luciferase of interest in target cells in a manner so that it is preferentially retained within the healthy cells but is either released from dead and dying cells or whose activity can be preferentially measured in dead and dying cells. We demonstrate that this assay is highly sensitive, specific, rapid, and can be performed in a single-step manner without the need for any expensive equipment. We further validate this assay by demonstrating its ability to detect cytotoxicity induced by several cellular and immune-therapeutic agents including antibodies, natural killer cells, chimeric antigen receptor expressing T cells and a bispecific T cell engager.

The cytotoxicities of three aconitum alkaloids- aconitine, hypaconitine and mesaconitine, and their abilities to bind DNA have been explored. Rat myocardial cells H9c2 were treated with aconitum alkaloids and assessed the cytotoxicities by using MTT assay and flow cytometry. Apoptosis was evidenced by the results of the annexin V/propidium iodide (PI) assay. Aconitine was found to be the most toxic in rat myocardial cells H9c2 in three aconitum alkaloids. At the same time, DNA adducts were isolated and then analyzed by UV-Vis spectroscopy after exposure to alkaloids, which indicated that three alkaloids could bind to DNA in rat myocardial cells H9c2. Furthermore, their binding modes were investigated by UV-Visible, fluorescence, DNA melting studies and ionic strength effect. Results indicated that the interaction between three alkaloids and DNA were intercalation coupled with electrostatic effect. The estimated binding constants were between 4.83 × 105 M−1 to 9.85 × 105 M−1 for three alkaloids at 298 K.

Avascular necrosis of the femur head (AVNFH) is a debilitating disease caused due to the use of alcohol, steroids, following trauma or unclear (idiopathic) etiology, affecting mostly the middle aged population. Clinically AVNFH is associated with impaired blood supply to the femoral head resulting in bone necrosis and collapse. Although Homocysteine (HC) has been implicated in AVNFH, levels of homocysteine and its associated pathway metabolites have not been characterized. We demonstrate elevated levels of homocysteine and concomitantly reduced levels of vitamins B6 and B12, in plasma of AVNFH patients. AVNFH patients also had elevated blood levels of sodium and creatinine, and reduced levels of random glucose and haemoglobin. Biophysical and ultrastructural analysis of AVNFH bone revealed increased remodelling and reduced bone mineral density portrayed by increased carbonate to phosphate ratio and decreased Phosphate to amide ratio together with disrupted trabeculae, loss of osteocytes, presence of calcified marrow, and elevated expression of osteocalcin in the osteoblasts localized in necrotic regions. Taken together, our studies for the first time characterize the metabolomic, pathophysiological and morphometric changes associated with AVNFH providing insights for development of new markers and therapeutic strategies for this debilitating disorder.

Elevated resting heart rate (RHR) predicts all-cause death. However, the relationship between RHR variation over years and mortality are still unknown. We aimed to analyze the association between RHR variation and all-cause mortality in the general population without cardiovascular diseases. A total of 46,873 subjects were included from the Kailuan Study (2006–2011). RHR readings were taken during three separate examinations and the RHR variation was defined using the standard deviation (RHR-SD) and the coefficient of variation. Participants were divided into four groups according to the quartiles of RHR-SD. All subjects were followed for a median of 49.4 months from the date of the 3rd examination to December 31, 2014. Up until the follow-up examinations, 973 (2.08%) participants had died. In a multivariate analysis, adjusting for variables potentially associated with death, the highest quartile of RHR-SD remained an independent predictor of all-cause mortality (Hazards ratio = 1.43, 95% confidence interval 1.18–1.74, P

To study the fractal dimensions of the soil particle size distributions (PSDs) within different plantations (of Pinus densiflora, Quercus acutissima, Robinia pseudoacacia, and Larix kaempferi) and evaluate PSDs as an indicator of the likelihood of Robinia pseudoacacia invasion, the soil porosity of 0–20 cm soil layers was measured at different plantations in the Yaoxiang Forest Farm, Shandong Province, China. The results showed that the fractal dimension (Dm) values varied from 2.59 to 2.70 among the different plantations and were significantly negatively correlated to sand content and positively correlated to silt content and clay content. Significant negative correlations were observed between Dm and both soil organic matter (SOM) (P

The main roles of equilibrative nucleoside transporters (ENTs) and concentrative nucleoside transporters (CNTs) are to transfer single nucleosides and analogues for the nucleic acid salvage pathway. Oligodeoxyribonucleotides (ODNs) can be transported into the cytoplasm or nucleus of cells under certain conditions. Among ODNs composed of a single type of nucleotide, the transport efficiency differs with the length and nucleotide composition of the ODNs and varies in different types of leukaemia cells; among the 5 tested random sequence ODNs and 3 aptamers with varying sequences, the data showed that some sequences were associated with significantly higher transport efficiency than others. The transport of ODNs was sodium, energy, and pH-independent, membrane protein-dependent, substrate nonspecific for ODNs and 4-nitrobenzylthioinosine (NBMPR)-insensitive, but it showed a low sensitivity to dipyridamole (IC50 = 35.44 µmol/L), distinguishing it from ENT1-4 and CNTs. The delivery efficiency of ODNs was superior to that of Lipofection and Nucleofection, demonstrating its potential applications in research or therapeutics. Moreover, this process was associated with p38 mitogen activated protein kinase (p38MAPK) instead of c-Jun N-terminal kinase (JNK) signalling pathways. We have denoted ODN transmembrane transport as equilibrative nucleic acid transport (ENAT). Overall, these findings indicate a new approach and mechanism for transmembrane transport of ODNs.

Serum bilirubin is a potent endogenous antioxidant and has been identified as cardiovascular risk in cohort studies, while the relation to type 2 diabetes (T2D) in the elderly remains unclear. We investigated both cross-sectional and prospective associations between serum bilirubin levels and T2D risk in the Dongfeng-Tongji (DFTJ) cohort, and replicated the prospective findings in a nested case-control study (509 cases and 509 controls) within the Singapore Chinese Health Study (SCHS). In the cross-sectional analysis of DFTJ cohort (15,575 participants with 2,532 diabetes cases), serum bilirubin levels (total, direct and indirect) increased in new on-set diabetes and decreased with the diabetic duration. In the longitudinal analysis of DFTJ cohort (772 incident diabetes cases during 4.5 years of follow-up among 12,530 diabetes-free participants at baseline), positive association was found between direct bilirubin and T2D risk comparing extreme quartiles, similar results were observed in the nested case-control study within SCHS. Total and indirect bilirubin levels were not significantly associated with T2D in either cohort. In conclusion, our findings do not support the protective association between serum bilirubin levels and incident T2D in the middle-aged and elderly adults; instead, direct bilirubin levels were associated with increased risk of T2D.

Current long term cryopreservation of cell stocks routinely requires the use of liquid nitrogen (LN2), because commonly used cryopreservation media containing cell membrane permeating cryoprotectants are thermally unstable when frozen at higher storage temperatures, e.g. −80 °C. This instability leads to ice recrystallization, causing progressive loss of cell viability over time under the storage conditions provided by most laboratory deep freezers. The dependency on LN2 for cell storage significantly increases operational expense and raises issues related to impaired working efficiency and safety. Here we demonstrate that addition of Ficoll 70 to cryoprotectant solutions significantly improves system thermal stability at the working temperature (~−80 °C) of laboratory deep freezers. Moreover, a medium comprised of Ficoll 70 and dimethyl sulfoxide (DMSO) in presence or absence of fetal bovine serum (FBS) can provide reliable cryopreservation of various kinds of human and porcine pluripotent stem cells at −80 °C for periods that extend up to at least one year, with the post-thaw viability, plating efficiency, and full retention of pluripotent phenotype comparable to that achieved with LN2 storage. These results illustrate the practicability of a promising long-term cryopreservation method that completely eliminates the need for LN2.

The present work aimed to investigate the activities and underlying mechanisms of dioscin against alcoholic liver fibrosis (ALF). In vivo liver fibrosis in mice was induced by an alcoholic liquid diet and in vitro studies were performed on activated HSC-T6 and LX2 cells treated with lipopolysaccharide. Our results showed that dioscin significantly attenuated hepatic stellate cells (HSCs) activation, improved collagen accumulation and attenuated inflammation through down-regulating the levels of myeloid differentiation factor 88 (MyD88), nuclear factor κB (NF-κB), interleukin (IL)-1, IL-6 and tumour necrosis factor-α by decreasing Toll-like receptor (TLR)4 expression both in vivo and in vitro. TLR4 overexpression was also decreased by dioscin, leading to the markedly down-regulated levels of MyD88, NF-κB, transforming growth factor-β1 (TGF-β1), α-smooth muscle actin (α-SMA) and type I collagen (COL1A1) in cultured HSCs. Suppression of cellular MyD88 by ST2825 or abrogation of NF-κB by pyrrolidine dithiocarbamate eliminated the inhibitory effects of dioscin on the levels of TGF-β1, α-SMA and COL1A1. In a word, dioscin exhibited potent effects against ALF via altering TLR4/MyD88/NF-κB signaling pathway, which provided novel insights into the mechanisms of this compound as an antifibrogenic candidate for the treatment of ALF in the future.

The mechanisms of the natural product dioscin against non-alcoholic fatty liver disease (NAFLD) are unclear. Thus, the purpose of the present study was to further confirm its effects of prevention and then to elucidate the potential mechanisms underlying its activity in mice. High-fat diet (HFD)-induced C57BL/6J mice and ob/ob mice were used as the experimental models. Serum and hepatic biochemical parameters were determined, and the mRNA and protein expression levels were detected. The results indicated that dioscin alleviated body weight and liver lipid accumulation symptoms, increased oxygen consumption and energy expenditure, and improved the levels of serum and hepatic biochemical parameters. Further investigations revealed that dioscin significantly attenuated oxidative damage, suppressed inflammation, inhibited triglyceride and cholesterol synthesis, promoted fatty acid β-oxidation, down-regulated MAPK phosphorylation levels, and induced autophagy to alleviate fatty liver conditions. Dioscin prevents diet induced obesity and NAFLD by increasing energy expenditure. This agent should be developed as a new candidate for obesity and NAFLD prevention.

We previously reported the promising effects of dioscin against liver injury, but its effect on liver fibrosis remains unknown. The present work investigated the activities of dioscin against liver fibrosis and the underlying molecular mechanisms. Dioscin effectively inhibited the cell viabilities of HSC-T6, LX-2 and primary rat hepatic stellate cells (HSCs), but not hepatocytes. Furthermore, dioscin markedly increased peroxisome proliferator activated receptor-γ (PPAR-γ) expression and significantly reduced a-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), collagen α1 (I) (COL1A1) and collagen α1 (III) (COL3A1) levels in vitro. Notably, dioscin inhibited HSCs activation and induced apoptosis in activated HSCs. In vivo, dioscin significantly improved body weight and hydroxylproline, laminin, α-SMA, TGF-β1, COL1A1 and COL3A1 levels, which were confirmed by histopathological assays. Dioscin facilitated matrix degradation and exhibited hepatoprotective effects through the attenuation of oxidative stress and inflammation, in addition to exerting anti-fibrotic effects through the modulation of the TGF-β1/Smad, Wnt/β-catenin, mitogen-activated protein kinase (MAPK) and mitochondrial signaling pathways, which triggered the senescence of activated HSCs. In conclusion, dioscin exhibited potent effects against liver fibrosis through the modulation of multiple targets and signaling pathways and should be developed as a novel candidate for the treatment of liver fibrosis in the future.

The mechanisms of the natural product dioscin against non-alcoholic fatty liver disease (NAFLD) are unclear. Thus, the purpose of the present study was to further confirm its effects of prevention and then to elucidate the potential mechanisms underlying its activity in mice. High-fat diet (HFD)-induced C57BL/6J mice and ob/ob mice were used as the experimental models. Serum and hepatic biochemical parameters were determined, and the mRNA and protein expression levels were detected. The results indicated that dioscin alleviated body weight and liver lipid accumulation symptoms, increased oxygen consumption and energy expenditure, and improved the levels of serum and hepatic biochemical parameters. Further investigations revealed that dioscin significantly attenuated oxidative damage, suppressed inflammation, inhibited triglyceride and cholesterol synthesis, promoted fatty acid β-oxidation, down-regulated MAPK phosphorylation levels, and induced autophagy to alleviate fatty liver conditions. Dioscin prevents diet induced obesity and NAFLD by increasing energy expenditure. This agent should be developed as a new candidate for obesity and NAFLD prevention.