Duhuo Jisheng Decoction suppresses apoptosis and mitochondrial dysfunction in human nucleus pulposus cells by miR-494/SIRT3/mitophagy signal axis.

BACKGROUND: Increasing evidence suggests that mitophagy is responsible for the pathogenesis of intervertebral disk (IVD) degeneration. Previous studies have shown that Duhuo Jisheng Decoction (DHJSD), a classic Fangji of traditional Chinese medicine, can delay IVD degeneration; however, its specific mechanism of action is unknown. In this study, we investigated the mechanism by which DHJSD treatment prevented IVD degeneration in IL-1ß-treated human nucleus pulposus (NP) cells in vitro. METHODS: Cell Counting Kit-8 was performed to explore the effects of DHJSD on the viability of NP cells exposed to IL-1ß. The mechanism by which DHJSD delays IVD degeneration was explored using luciferase reporter assay, RT-qPCR, western blotting, TUNEL assay, mitophagy detection assay, Mito-SOX, Mitotracker and in situ hybridization. RESULTS: We observed that DHJSD enhanced the viability of NP cells treated with IL-1ß in a concentration-time dependent approach. Moreover, DHJSD lessened IL-1ß-induced NP apoptosis and mitochondrial dysfunction and activated mitophagy in NP cells treated with IL-1ß. Mitophagy suppressor cyclosporin A reversed the beneficial impacts of DHJSD in NP cells. In addition, the differential expression of miR-494 regulated IL-1ß-induced NP apoptosis and mitochondrial dysfunction, and the protective impact of miR-494 on NP cells treated with IL-1ß was achieved by mitophagy activation, which was regulated by its target gene, sirtuin 3 (SIRT3). Finally, we observed that DHJSD treatment could effectively delay IL-1ß-induced NP apoptosis by affecting the miR-494/SIRT3/mitophagy signal axis. CONCLUSIONS: These results show that the miR-494/SIRT3/mitophagy signaling pathway is responsible for the apoptosis and mitochondrial dysfunction of NP cells and that DHJSD may exert protective effects against IVD degeneration by regulating the miR-494/SIRT3/mitophagy signal axis.

Duhuo jisheng decoction suppresses matrix degradation and apoptosis in human nucleus pulposus cells and ameliorates disc degeneration in a rat model.

ETHNOPHARMACOLOGICAL RELEVANCE: The lower back pain (LBP) caused by intervertebral disc (IVD) degeneration brings a heavy burden to society. A classic treatment method of Chinese medicine, fangji-duhuo jisheng decoction (DHJSD), has been effective in the clinical treatment of LBP, although the underlying mechanism remains unknown. AIM OF THE STUDY: In this work, the main objective was to study the effects of DHJSD on in vitro IVD degeneration of human nucleus pulposus (NP) cells after pressure treatment and on an in vivo interrupted IVD degeneration rat model. MATERIALS AND METHODS: The effects of DHJSD on the viability of NP cells were detected using Cell Counting Kit-8. RT-qPCR, western blotting, TUNEL assay, transmission electron microscopy, and immunofluorescence staining were performed to explore the molecular mechanism underlying protection against compression-induced matrix degradation and apoptosis in NP cells by DHJSD. Furthermore, the effects of DHJSD on IVD degeneration in a rat IDD model were also determined. RESULTS: We found that DHJSD increased the viability of NP cells in a concentration- and time-dependent manner. Furthermore, DHJSD significantly reduced compression-induced NP matrix degeneration and apoptosis, activated autophagy, and inhibited the p38/MAPK signaling pathway in NP cells subjected to compression. Autophagy inhibitor 3-MA and p38/MAPK signaling pathway activator anisomycin reversed the beneficial effects of DHJSD in NP cells, indicating that DHJSD protects against IVD degeneration by autophagy activation and P38/MAPK signaling pathway inhibition. Furthermore, DHJSD treatment effectively delayed IVD degeneration in a puncture-induced IDD rat model. CONCLUSIONS: DHJSD prevents compression-induced matrix degradation and cell apoptosis through regulating autophagy and the P38/MAPK signaling pathway. The mechanism underlying the effects of DHSJD elucidated in this study provides a new direction for LBP treatment.

The Influence of Audio-Visual Interactions on Psychological Responses of Young People in Urban Green Areas: A Case Study in Two Parks in China.

Audio-visual interactions in green spaces are important for mental health and wellbeing. However, the influence of audio-visual interactions on psychological responses is still less clear. This study introduced a new method, namely the audio-visual walk (AV-walk), to obtain data on the audio-visual context, audio-visual experiences, and psychological responses in two typical parks, namely Cloves Park and Music Park in Harbin, China. Some interesting results are as follows: First, based on Pearson's correlation analysis, sound pressure level and roughness were significantly correlated with psychological responses in Cloves Park (p < 0.05). Second, the results of stepwise regression models showed the impact intensity of acoustic comfort was 1.64-1.68 times higher than that of visual comfort on psychological responses of emotion dimension, while visual comfort was 1.35-1.37 times higher than acoustic comfort on psychological responses of cognition dimension in Music Park. In addition, an orthogonal analysis diagram explained the influence of audio-visual interactions on psychological responses of young people. The audio-visual context located beside the waterscape with a relatively higher level of acoustic and visual comfort was the most cheerful (2.60), relaxed (2.45), and energetic (2.05), while the audio-visual context close to an urban built environment tended to be both acoustically and visually uncomfortable, and the psychological state was decreased to the most depressed (-0.25), anxious (-0.75), fatigued (-1.13) and distracted (-1.13).

Exogenous kallikrein protects against diabetic nephropathy.

The kallikrein-kinin system has been shown to be involved in the development of diabetic nephropathy, but specific mechanisms are not fully understood. Here, we determined the renal-protective role of exogenous pancreatic kallikrein in diabetic mice and studied potential mechanisms in db/db type 2 diabetic and streptozotocin-induced type 1 diabetic mice. After the onset of diabetes, mice were treated with either pancreatic kallikrein (db/db+kallikrein, streptozotocin+kallikrein) or saline (db/db+saline, streptozotocin+saline) for 16 weeks, while another group of streptozotocin-induced diabetic mice received the same treatment after onset of albuminuria (streptozotocin'+kallikrein, streptozotocin'+saline). Db/m littermates or wild type mice were used as non-diabetic controls. Pancreatic kallikrein had no effects on body weight, blood glucose and blood pressure, but significantly reduced albuminuria among all three groups. Pathological analysis showed that exogenous kallikrein decreased the thickness of the glomerular basement membrane, protected against the effacement of foot process, the loss of endothelial fenestrae, and prevented the loss of podocytes in diabetic mice. Renal fibrosis, inflammation and oxidative stress were reduced in kallikrein-treated mice compared to diabetic controls. The expression of kininogen1, tissue kallikrein, kinin B1 and B2 receptors were all increased in the kallikrein-treated compared to saline-treated mice. Thus, exogenous pancreatic kallikrein both prevented and ameliorated diabetic nephropathy, which may be mediated by activating the kallikrein-kinin system.

Investigating the tool marks on oracle bones inscriptions from the Yinxu site (ca., 1319-1046 BC), Henan province, China.

Oracle Bone Inscriptions in the Shang dynasty (1600-1046 BC) are the earliest well-developed writing forms of the Chinese character system, and their carving techniques have not been studied by tool marks analysis with microscopy. In this study, a digital microscope with three-dimensional surface reconstruction based on extended depth of focus technology was used to investigate tool marks on the surface of four pieces of oracle bones excavated at the eastern area of Huayuanzhuang, Yinxu site(ca., 1319-1046 BC), the last capital of the Shang dynasty, Henan province, China. The results show that there were two procedures to carve the characters on the analyzed tortoise shells. The first procedure was direct carving. The second was "outlining design," which means to engrave a formal character after engraving a draft with a pointed tool. Most of the strokes developed by an engraver do not overlap the smaller draft, which implies that the outlining design would be a sound way to avoid errors such as wrong and missing characters. The strokes of these characters have different shape at two ends and variations on width and depth of the grooves. Moreover, the bottom of the grooves is always rugged. Thus, the use of rotary wheel-cutting tools could be ruled out. In most cases, the starting points of the strokes are round or flat while the finishing points are always pointed. Moreover, the strokes should be engraved from top to bottom. When vertical or horizontal strokes had been engraved, the shell would be turned about 90 degrees to engrave the crossed strokes from top to bottom. There was no preferred order to engrave vertical or horizontal strokes. Since both sides of the grooves of the characters are neat and there exists no unorganized tool marks, then it is suggested that some sharp tools had been used for engraving characters on the shells. Microsc. Res. Tech. 79:827-832, 2016. © 2016 Wiley Periodicals, Inc.

De novo assembly and transcriptome analysis of sclerotial development in Wolfiporia cocos.

Wolfiporia cocos Ryvarden et Gilbertson, a well-known medicinal fungus in the Basidiomycetes, is widely distributed in East Asia. Its dried sclerotium, which is known as Fuling in China, has been used as a traditional crude drug in Chinese traditional medicine for thousand years. However, little is known about how the sclerotium is developed at the genetic level. In this study, the de novo sequencing of sclerotia of W. cocos (S1_initial stage; S2_developmental stage and S3_mature stage) was carried out by illumina HiSeq 2000 technology. 27,438 unigenes were assembled from ~30Gbp raw data, and 12,093 unigenes were significantly annotated. The analysis of expression profiles during development returned 304 differentially expressed genes (DEGs), which were clustered into four different groups according to their expression trends. Especially for the maturation stage (S3), the sclerotium exhibited a markedly different expression profile from other stages. We further showed that peroxisome, unsaturation of fatty acids and degradation pathway were respectively prevalent in S1, S2 and S3 stages as evidenced by enrichment analysis. To our knowledge, this study represents the first report of sclerotial development transcriptomics in W. cocos. The obtained results provide novel insights into the developmental biology of the sclerotia, which is helpful for future studies about cultivation and breeding of W. cocos.

Polysaccharides from Rhizoma Panacis Majoris and its anti-oxidant activity.

Response surface method (RSM) was employed to optimize the extraction conditions of polysaccharides from Rhizoma Panacis Majoris (the rhizomes of Panax japonicus C. A. Mey. var. major (Burk.) C. Y. Wu et K. M. Feng) (RPMP), a well-known Chinese traditional medicine. In order to obtain the optimal processing parameters, a three-variable Box-Behnken designs (BBD) were applied for experimental designs. RSM analysis indicated the good correspondence between experimental and predicted values, the optimal conditions for the yield of polysaccharides were as follows: the ultrasound time is 31.15 min, extraction temperature is 92.50 °C, and the ratio of water to raw material is 40 mL/g. The maximum value (13.87 ± 0.16%) of the yield of polysaccharides was obtained under these optimal conditions. The molecular weight (MW) was determined to be 1.48 × 10(5)(±0.39%)Da by HPSEC-MALLS-RID chromatography system. FT-IR spectra demonstrated obvious characteristic peaks of polysaccharides. The antioxidant activities of RPMP were investigated including scavenging activity of hydrogen radicals, ABTS radicals, and free radicals of superoxide anion in vitro, and the results exhibited that RPMP had a good potential for antioxidant.

Nitrogen-doped porous carbon derived from biomass waste for high-performance supercapacitor.

High capacitance property and low cost are the pivotal requirements for practical application of supercapacitor. In this paper, a low cost and high capacitance property nitrogen-doped porous carbon with high specific capacitance is prepared. The as-prepared nitrogen-doped porous carbon employing potato waste residue (PWR) as the carbon source, zinc chloride (ZnCl2) as the activating agent and melamine as nitrogen doping agent. The morphology and structure of the carbon materials are studied by scanning electron microscopy (SEM), N2 adsorption/desorption, X-ray diffraction (XRD) and Raman spectra. The surface area of the nitrogen-doped carbon which prepared under 700°C is found to be 1052m(2)/g, and the specific capacitance as high as 255Fg(-1) in 2M KOH electrolyte is obtained utilize the carbon as electrode materials. The electrode materials also show excellent cyclability with 93.7% coulombic efficiency at 5Ag(-1) current density of for 5000cycles.

Peak systolic velocity of superior thyroid artery for the differential diagnosis of thyrotoxicosis.

AIM: The differentiation of destruction-induced thyrotoxicosis and Graves' disease (GD) is of great importance for selection of proper therapy. Radioactive iodine uptake (RAIU) is the gold standard for differentiating these two conditions but its application has remained somewhat limited. Thyroid color Doppler flow sonography (CDFS) is a potential alternative of RAIU but more supporting evidence is warranted. In the present study, a standard operative procedure was developed to measure the mean peak systolic velocity of superior thyroid artery (STA-PSV) and evaluate its role in the differential diagnosis of thyrotoxicosis. METHODS: A total of 135 patients with untreated thyrotoxicosis were enrolled into one retrospective study (GD, n = 103; thyroiditis, n = 32) and another prospective study recruited 169 patients (GD, n = 118; thyroiditis, n = 51). Thirty normal controls were also enrolled. Thyroid function, anti-TSH-receptor antibody (TRAb), RAIU, CFDS of thyroid and STA-PSV were performed for each patient. Receiver operator curve (ROC) was used to evaluate the diagnostic value of STA-PSV in a retrospective study so as to seek the optimal cutoff point. Then the cutoff point value was used to validate its diagnostic value in a prospective study and in another thyrotoxicosis population. RESULTS: STA-PSV of GD was significantly higher than that of thyroiditis in both retrospective and prospective studies. The area under the ROC curve of mean STA-PSV was 0.8799 and 0.9447 in the retrospective and prospective studies respectively. If a mean STA-PSV cutoff point of 50.5 cm/s was set from the retrospective analysis for the prospective study, the sensitivity and specificity in distinguishing GD from thyroiditis were 81.04% and 96.08% respectively. Mean STA-PSV and TRAb had similar area under ROC. The coefficients of variation in STA-PSV measurement were lower than 10% for the euthyroid, thyroiditis and GD groups. CONCLUSIONS: STA-PSV is a feasible supplement alternative of RAIU for differentiating the causes of thyrotoxicosis.

Tetramethylpyrazine protects palmitate-induced oxidative damage and mitochondrial dysfunction in C2C12 myotubes.

AIMS: Tetramethylpyrazine (TMP), one of the active ingredients isolated from a Chinese herbal prescription, possesses protective effects against oxidative stress caused by high glucose in endothelial cells. In this study, the role of TMP in preventing muscle cells from palmitate-induced oxidative damage was investigated and the possible mechanisms of action elucidated. MAIN METHODS: Mitochondrial reactive oxygen species (ROS) were measured in C2C12 myotubes, a palmitate-induced oxidative stress cell model, with or without TMP. Both mitochondrial membrane potential (MMP) and oxygen consumption were assessed in conjunction with quantification of mitochondrial DNA and mitochondrial biogenesis-related factors, such as peroxisome proliferator-activated receptor-γ coactivator 1 α (PGC1α), nuclear respiratory factor 1 (NRF1) and mitochondrial transcription factor A (Tfam), by real-time polymerase chain reaction. Expression of mitochondrial respiratory chain complex III as an index of mitochondrial function was evaluated by immunoblotting, and glucose transport into the C2C12 myotube examined by analyzing 2-deoxy-[(3)H]glucose uptake. KEY FINDINGS: TMP significantly alleviated palmitate-induced mitochondrial ROS production, mitigated mitochondrial dysfunction and increased D-loop mRNA expression as compared with the control. This was accompanied by a marked reversal of palmitate-induced down-regulation in the expression of mitochondrial biogenesis-related factors (PGC1α, NRF1 and Tfam) and decreased glucose uptake in C2C12 myotubes. As a result, cell respiration, as reflected by the elevated expression of mitochondrial respiratory chain complex III and oxygen consumption, was enhanced. SIGNIFICANCE: TMP is capable of protecting C2C12 myotubes against palmitate-induced oxidative damage and mitochondrial dysfunction, and improving glucose uptake in muscle cells partially through the up-regulation of mitochondrial biogenesis.