Molecular mechanisms of the Guizhi decoction on osteoarthritis based on an integrated network pharmacology and RNA sequencing approach with experimental validation

Background: Our aim was to determine the potential pharmacological mechanisms of the Guizhi decoction (GZD) in the treatment of osteoarthritis (OA) through an integrated approach of network pharmacological analyses, RNA sequencing (RNA-seq), and experimental validation.Methods: The quality control and identification of bioactive compounds of the GZD were carried out by using ultra-performance liquid chromatography (UPLC), and their OA-related genes were identified through overlapping traditional Chinese medicine systems pharmacology database (TCMSP), DrugBank and SEA Search Server databases, and GeneCards. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were implemented after constructing the component–target network. RNA-seq was used to screen differentially expressed genes (DEGs) under intervention conditions with and without the GZD in vitro. The crossover signaling pathways between RNA-seq and network pharmacology were then analyzed. Accordingly, protein–protein interaction (PPI) networks, GO, and KEGG analysis were performed using the Cytoscape, STRING, or DAVID database. The OA rat model was established to further verify the pharmacological effects in vivo. Hematoxylin–eosin (H&E) and safranin O/fast green (S-O) staining were used to grade the histopathological features of the cartilage. We verified the mRNA and protein expressions of the key targets related to the TNF signaling pathways in vivo and in vitro by qPCR, Western blotting (WB), and immunofluorescence assay. In addition, we also detected inflammatory cytokines in the rat serum by Luminex liquid suspension chip, which included tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β).Results: Eighteen compounds and 373 targets of the GZD were identified. A total of 2,356 OA-related genes were obtained from the GeneCards database. A total of three hub active ingredients of quercetin, kaempferol, and beta-sitosterol were determined, while 166 target genes associated with OA were finally overlapped. The RNA-seq analysis revealed 1,426 DEGs. In the KEGG intersection between network pharmacology and RNA-seq analysis, the closest screening relevant to GZD treatment was the TNF signaling pathway, of which TNF, IL-6, and IL-1β were classified as hub genes. In consistent, H&E and S-O staining of the rat model showed that GZD could attenuate cartilage degradation. When compared with the OA group in vivo and in vitro, the mRNA levels of TNF-α, IL-1β, IL-6, matrix metalloproteinase 3 (MMP3), and matrix metalloproteinase 9 (MMP9) were all downregulated in the GZD group (all p < 0.05). The expression levels of anabolic proteins (Col2α1 and SOX9) were all higher in the GZD group than in the OA group (p < 0.05), while the expression levels of the catabolic proteins (MMP9 and COX-2) and TNF-α in the GZD group were significantly lower than those in the OA group (p < 0.05). In addition, the expression levels of TNF, IL-6, and IL-1β were upregulated in the OA group, while the GZD group prevented such aberrations (p < 0.01).Conclusion: The present study reveals that the mechanism of the GZD against OA may be related to the regulation of the TNF signaling pathway and inhibition of inflammatory response

Traditional Chinese Herbal Patch for Short-Term Management of Knee Osteoarthritis: A Randomized, Double-Blind, Placebo-Controlled Trial

Objective. To assess the short-term efficacy and safety of two kinds of Traditional Chinese herbal patches, Fufang Nanxing Zhitong Gao (FNZG) and Shangshi Jietong Gao (SJG), for painful knee osteoarthritis (OA). Methods. Patients were randomly enrolled in a double-blind, placebo-controlled study to receive FNZG (n=60), SJG (n=60), or placebo patch (n=30) for 7 days. Outcome measures included visual analogue scale (VAS), Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), and Traditional Chinese Medicine Syndrome Questionnaire (TCMSQ) subscale. Results. Although there was no significant difference among, three groups in short-term pain management, patients receiving FNZG got significant improvement in symptom of fear of coldness as compared with placebo patch (P=0.029). The most common local adverse events of rash, itching, erythema, and slightly damaged skin were observed in 7% of participants. Conclusions. FNZG may be a useful treatment for symptom of knee OA and merits long-term study in broader populations

Effectiveness, Medication Patterns, and Adverse Events of Traditional Chinese Herbal Patches for Osteoarthritis: A Systematic Review

Objective. The aim of this study is to systematically evaluate the evidence whether traditional Chinese herbal patches (TCHPs) for osteoarthritis (OA) are effective and safe and analyze their medication patterns. Methods. A systematic literature search was performed using all the possible Medical Subject Headings (MeSH) and keywords from January 1979 to July 2013. Both randomized controlled trials (RCTs) and observational studies were included. Estimated effects were analyzed using mean difference (MD) or relative risk (RR) with 95% confidence intervals (CI) and meta-analysis. Results. 86 kinds of TCHPs were identified. RCTs and controlled clinical trials (CCTs) which were mostly of low quality favored TCHPs for local pain and dysfunction relief. TCHPs, compared with diclofenac ointment, had significant effects on global effectiveness rate (RR = 0.50; 95% CI (0.29, 0.87)). Components of formulae were mainly based on the compounds “Xiao Huo Luo Dan” (Minor collateral-freeing pill) and “Du Huo Ji Sheng Tang” (Angelicae Pubescentis and Loranthi decoction). Ten kinds of adverse events (AEs), mainly consisting of itching and/or local skin rashes, were identified after 3-4 weeks of follow-up. Conclusions. TCHPs have certain evidence in improving global effectiveness rate for OA; however, more rigorous studies are warranted to support their use

Static voltage sharing technology of multi-break mechanical switch for hybrid HVDC breaker

The hybrid high-voltage direct current (HVDC) breaker combines mechanical and power electronics switching that enables it to interrupt power flows within a few milliseconds. Mechanical switch is a key component of hybrid HVDC breaker and has a number of serially connected interrupter units which ideally would divide the voltage equally. The static voltage distribution characteristics and voltage sharing design of a multi-break mechanical switch were discussed in this study. A finite-element model was developed to study the static voltage distribution characteristics and capacitance parameters of multi-break mechanical switch (which actually consists of resistance and capacitance parameters under direct current) as a preliminary study. Comparisons were made under the simulation of vertical and U-shaped arrangement forms. The results indicate that the static voltage distribution of the high-voltage terminal is at least more than 65%, whereas the severe non-uniform voltage distribution can be well improved by means of the method proposed in this study

Discharge voltage prediction of UHV AC transmission line–tower air gaps by a machine learning model

Full-scale discharge tests of transmission line–tower air gaps are costly and time-consuming, and they cannot exhaustively simulate all the gap configurations in practical engineering. In this paper, a machine learning model established by support vector machine is introduced to predict the switching impulse discharge voltages of the ultra-high-voltage (UHV) AC transmission line–tower air gaps. The three-dimensional finite element models of a UHV cup-type tower and a UHV compact transmission line were established for electric field calculation, and some features were extracted from the hypothetic discharge channel and the shortest path between the bundled conductor and the tower. These features under a given voltage were normalised and input to the SVM model, while the output is two binary values, respectively, representing gap withstanding or breakdown. Trained by experimental data of one type of the UHV transmission line–tower gaps, the SVM model is able to predict the discharge voltages of another gap type. The mean absolute percentage errors of the two engineering gap types, under different gap distances, are 8.31 and 4.86%, respectively, which are acceptable for engineering applications. The results provide a possible way to obtain the discharge voltages of complicated engineering gaps by mathematical calculations

Electric field calculation and grading ring design for 750 kV four-circuits transmission line on the same tower with six cross-arms

The transmission corridors can be effectively saved using 750 kV four-circuit transmission line on the same tower. In order to reduce the corona level, structure optimisation of fittings plays an important role in the transmission line design. In this study, the electric field distribution of 750 kV transmission line on six cross-arms tower was calculated based on finite element method, and the influences of pipe diameter, ring diameter and installation position of the grading ring were analysed. The results show that the increase of pipe diameter can reduce the electric field strength of grading ring while the increase of installation position and decrease of ring diameter are important to protect the end fitting of composite insulator. When using grading ring with the pipe diameter of 100 mm, the ring diameter of 700–900 mm and the installation position of 150–250 mm, the electric field strength can be controlled in a reasonable range. These results can provide references for the grading ring design of 750 kV four-circuit transmission line on the same tower

Data Mining in the Vibration Signal of the Trip Mechanism in Circuit Breakers Based on VMD-PSR

To address the difficulty in characterizing early mechanical faults in the trip mechanism of circuit breakers, a data mining method based on variational mode decomposition (VMD) and phase space reconstruction (PSR) method was proposed. First, the vibration signal in the trip stage was separated from the whole according to the current features. Then, it was decomposed using the VMD algorithm to obtain the intrinsic mode functions (IMFs) and these sub signals were mapped to high-dimensional phase space based on the PSR algorithm. Then, the features of the attractor trail shape and the recurrence plot matrixes were extracted. In order to judge the fault in the trip mechanism, a fault simulation test was carried out and the characteristic under different faults was analyzed. Based on these samples, a fault identification model is established by support vector machine (SVM) and the effectiveness is verified by other test samples. The accuracy of the SVM model is 98%, which is higher than that of the BPNN and KNN clustering models. This research supplements the existing method for condition evaluation of the trip mechanism and can provide a reference for circuit breaker fault diagnosis

Fermentation of increasing ratios of grain starch and straw fiber: effects on hydrogen allocation and methanogenesis through in vitro ruminal batch culture

Grain starch has a faster rate of rumen fermentation than straw fiber and causes a rapid increase in ruminal molecular hydrogen (H2) partial pressure, which may promote other H2 sinks to compete H2 away from methanogenesis. The study was designed to investigate the effects of increasing ratios of grain starch to straw fiber on hydrogen allocation and methanogenesis through in vitro ruminal batch incubation. Corn grain and corn straw were employed as starch and fiber source respectively. Seven treatments were the ratios of corn grain to corn straw (RGS) being 0:6, 1:5, 2:4, 3:3, 4:2, 5:1, and 6:0. Elevating RGS increased dry matter (DM) degradation and decreased methane (CH4) and hydrogen gas (gH2) production relative to DM degraded. Elevating RGS increased volatile fatty acid (VFA) concentration, propionate molar percentage and microbial protein (MCP) concentration, decreased acetate molar percentage, acetate to propionate ratio and estimated net metabolic hydrogen ([H]) production relative to DM degraded. Elevating RGS decreased the molar percentage of [H] utilized for CH4 and gH2 production. In summary, increasing ratios of grain starch to straw fiber altered rumen fermentation pathway from acetate to propionate production, reduced the efficiency of [H] production with the enhancement of MCP synthesis, and led to a reduction in the efficiency of CH4 and gH2 production

Osthole Inhibits Proliferation and Induces Catabolism in Rat Chondrocytes and Cartilage Tissue

Background/Aims: Cartilage destruction is thought to be the major mediator of osteoarthritis. Recent studies suggest that inhibition of subchrondral bone loss by anti-osteoporosis (OP) drug can protect cartilige erosion. Osthole, as a promising agent for treating osteoporosis, may show potential in treating osteoarthritis. The purpose of this study was to investigate whether Osthole affects the proliferation and catabolism of rat chondrocytes, and the degeneration of cartilage explants. Methods: Rat chondrocytes were treated with Osthole (0 μM, 6.25 μM, 12.5 μM, and 25 μM) with or without IL1-β (10ng/ml) for 24 hours. The expression levels of type II collagen and MMP13 were detected by western Blot. Marker genes for chondrocytes (A-can and Sox9), matrix metalloproteinases (MMPs), aggrecanases (ADAMTS5) and genes implicated in extracellular matrix catabolism were evaluated by qPCR. Cell proliferation was assessed by measuring proliferating cell nuclear antigen (PCNA) expression and fluorescence activated cell sorter. Wnt7b/β-catenin signaling was also investigated. Cartilage explants from two-week old SD rats were cultured with IL-1β, Osthole and Osthole plus IL-1β for four days and glycosaminoglycan (GAG) synthesis was assessed with toluidine blue staining and Safranine O/Fast Green FCF staining, collagen type II expression was detected by immunofuorescence. Results: Osthole reduced expression of chondrocyte markers and increased expression of MMP13, ADAMTS5 and MMP9 in a dose-dependent manner. Catabolic gene expression levels were further improved by Osthole plus IL-1β. Osthole inhibited chondrocyte proliferation. GAG synthesis and type II collagen were decreased in both the IL-1β groups and the Osthole groups, and significantly reduced by Osthole plus IL-1β. Conclusions: Our data suggested that Osthole increases the catabolism of rat chondrocytes and cartilage explants, this effect might be mediated through inhibiting Wnt7b/β-catenin pathway

An Inhibitory Role of Osthole in Rat MSCs Osteogenic Differentiation and Proliferation via Wnt/β-Catenin and Erk1/2-MAPK Pathways

Background/Aims: Bone marrow-derived mesenchymal stem cells (MSCs) are responsible for new bone formation during adulthood. Accumulating evidences showed that Osthole promotes the osteogenic differentiation in primary osteoblasts. The aim of this study was to investigate whether Osthole exhibits a potential to stimulate the osteogenic differentiation of MSCs and the underlying mechanism. Methods: MSCs were treated with a gradient concentration of Osthole (6.25 µM, 12.5 µM, and 25 µM). Cell proliferation was assessed by western blotting with the proliferating cell nuclear antigen (PCNA) and Cyclin D1 antibodies, fluorescence activated cell sorting (FACS), and cell counting kit 8 (CCK8). MSCs were cultured in osteogenesis-induced medium for one or two weeks. The osteogenic differentiation of MSCs was estimated by Alkaline Phosphatase (ALP) staining, Alizarin red staining, Calcium influx, and quantitative PCR (qPCR). The underlying mechanism of Osthole-induced osteogenesis was further evaluated by western blotting with antibodies in Wnt/β-catenin, PI3K/Akt, BMPs/smad1/5/8, and MAPK signaling pathways. Results: Osthole inhibited proliferation of rat MSCs in a dose-dependent manner. Osthole suppressed osteogenic differentiation of rat MSCs by down-regulating the activities of Wnt/β-catenin and Erk1/2-MAPK signaling. Conclusions: Osthole inhibits the proliferation and osteogenic differentiation of rat MSCs, which might be mediated through blocking the Wnt/β-catenin and Erk1/2-MAPK signaling pathways