Paeoniflorin, the Main Monomer Component of Paeonia lactiflora, Exhibits Anti-inflammatory Properties in Osteoarthritis Synovial Inflammation.

OBJECTIVE: To explore the mechanism of paeoniflorin (PF) on osteoarthritis (OA) synovial inflammation from network pharmacology to experimental pharmacology. METHODS: Targets of OA were constructed by detecting the database of network database platforms (Therapeutic Target database, DrugBank and GeneCards), and the targets of PF were constructed by PubChem and Herbal Ingredients' Targets database. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of these co-targeted genes were conducted via Database for Annotation, Visualization, and Integrated Discovery (DAVID) database, and protein-protein interaction (PPI) networks were conducted via the search tool for the retrieval of interacting genes (STRING) database. Cell counting kit-8 (CCK-8) assay was performed to assess the potential toxicity of PF on human OA fibroblast-like synoviocytes (FLS), quantitative real-time polymerase chain reaction (qPCR), enzyme-linked immunosorbent assay (ELISA) and Western blot were used to verify the potential mechanism of PF in synovial inflammation. RESULTS: Twenty-six co-targeted genes were identified. GO enrichment results showed that these co-targeted genes were most likely localized in the cytoplasm, and the biological processes mainly involved 'cellular response to hypoxia' 'lipopolysaccharide (LPS)-mediated signaling pathway' and 'positive regulation of gene expression'. KEGG pathway analysis indicated that these co-targeted genes may function through pathways associated with 'hypoxia-inducible factor-1 (HIF-1) signaling pathway' and 'tumor-necrosis factor (TNF) signaling pathway'. The PPI network showed that the top 3 hub genes were TP53, TNF, and CASP3. Molecular docking results showed that PF was well docking with TNF. CCK-8 showed no potential toxicity of 10, 20 and 50 µmol/L PF on human OA FLS. And PF significantly decreased the expression levels of interleukin-1 ß, interleukin-6, TNF-α matrix metalloproteinase 13 (MMP13), and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5) and TNF-α in LPS-induced OA FLS. CONCLUSION: PF exhibited potent anti-inflammatory effect in OA synovial inflammation.

Antioxidant Properties of Triterpenoids Isolated from Bagasse-Cultivated Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), at Different Developmental Stages.

Although triterpenoids are one of the main active ingredients in Ganoderma lucidum, their accumulation and antioxidant activity during the different developmental stages of G. lucidum cultivation in bagasse remains unclear. In this study, we investigated the content and antioxidant activity of total triterpenoids extracted from G. lucidum strain GL102 during the five growth stages. The obtained results showed that the highest content (12.06 mg/g) was detected in stage 3 (young fruiting body), similar to the contents of ganoderic acids B and G. However, ganoderic acids A and D exhibited maximal contents in stage 5 (spore-ejected fruiting body). The triterpenoids extracted during stage 5 were most capable of scavenging DPPH, OH, and ABTS(+) radicals, with scavenging rates of 65.88%, 86.45%, and 97.91%, respectively. Based on in vivo antioxidant assays conducted on zebrafish, the safe concentration of these triterpenoids was 0.03 mg/mL. At this concentration, the G. lucidum triterpenoids extracted during stage 5 could decrease lipopolysaccharide-induced intracellular reactive oxygen species to a level that was nearly normal (similar to the control group). The accumulation profile and antioxidant activity results reported herein provide the scientific basis needed to promote the utilization of triterpenoids derived from bagasse-cultivated G. lucidum.

Cinnamic Aldehyde, the main monomer component of Cinnamon, exhibits anti-inflammatory property in OA synovial fibroblasts via TLR4/MyD88 pathway.

Cinnamon is a wildly used traditional Chinese herbal medicine for osteoarthritis (OA) treatment, but the underlying mechanism remains ambiguous. The purpose of this study is to explore the mechanism of cinnamic aldehyde (CA), a bioactive substance extracted from Cinnamon, on synovial inflammation in OA. A total of 144 CA-OA co-targeted genes were identified by detect databases (PubChem, HIT, TCMSP, TTD, DrugBank and GeneCards). The results of GO enrichment analysis indicated that these co-targeted genes have participated in many biological processes including 'inflammatory response', 'cellular response to lipopolysaccharide', 'response to drug', 'immune response', 'lipopolysaccharide-mediated signalling pathway', etc. KEGG pathway analysis showed these co-targeted genes were mainly enriched in 'Toll-like receptor signalling pathway', 'TNF signalling pathway', 'NF-kappa B signalling pathway', etc. Molecular docking demonstrated that CA could successfully bind to TLR2 and TLR4. The results of in vitro experiments showed no potential toxicity of 10, 20 and 50 µM/L CA on human OA FLS, and CA can significantly inhibit the inflammation in LPS-induced human FLS. Further experimental mechanism evidence confirmed CA can inhibited the inflammation in LPS-induced human OA FLS via blocking the TLR4/MyD88 signalling pathway. Our results demonstrated that CA exhibited strong anti-inflammation effect in OA FLS through blocking the activation of TLR4/MyD88 signalling pathway, suggesting its potential as a hopeful candidate for the development of novel agents for the treatment of OA.

Effect of Tai Chi exercise on lower limb function and balance ability in patients with knee osteoarthritis: A protocol of a randomized controlled trial.

BACKGROUND: Knee osteoarthritis (KOA) is a chronic degenerative joint disease commonly occurring in middle-aged and elderly people. The main clinical manifestations are joint pain, limited activity, and decreased muscle strength resulting in decreased motor control ability. Exercise therapy is an effective method to enhance muscle strength of lower limbs, while China's traditional skill Tai Chi (TC) is a combination of activity and inertia, internal and external exercise therapy. In recent years, scholars at home and abroad have found that regular TC can effectively improve patients' lower limb function and balance ability. The purpose of this study is to explore the effects of TC on lower limb function and balance ability in patients with KOA. METHODS: This is a prospective randomized controlled clinical trial. One hundred forty-six cases of KOA patients will be randomly divided into experimental group and control group according to 1:1 ratio, 73 cases in each group, the control group: sodium hyaluronate; experimental group: TC added on the basis of the control group. Both groups will receive standard treatment for 5 weeks and will be followed up for 3 months. Observation indicators include: the western Ontario and McMaster universities osteoarthritis index; hospital for special surgery knee score; balance stability index, liver and kidney function, adverse reaction rate, etc. SPSS 23.0 software will be used for data analysis. DISCUSSION: This study will evaluate the effects of TC on lower limb function and balance ability of patients with KOA. The results of this trial will provide a clinical basis for the selection of exercise therapy for patients with KOA.

Efficacy and Mechanism of Electroacupuncture Treatment of Rabbits With Different Degrees of Knee Osteoarthritis: A Study Based on Synovial Innate Immune Response.

Knee osteoarthritis (KOA) is a chronic degenerative bone and joint disease, which is often clinically manifested as pain, joint swelling, and deformity. Its pathological manifestations are mainly synovial inflammation and cartilage degeneration. This study aims to investigate the efficacy of electro-acupuncture (EA) on model rabbits with varying degrees of KOA and to study the mechanism of EA on KOA based on the innate immune response. Mild and moderate rabbit KOA models were established using a modified Hluth method, and EA was given to both the mild and moderate model groups. The Lequesne-MG index was used to evaluate the behavioral changes in the rabbits before and after EA treatment. Morphological changes in the synovial membrane and cartilage of each group were observed by H&E staining. The Mankin scoring standard and the Krenn scoring standard were used to score the pathology of the cartilage tissue and synovial tissue, respectively. The inflammatory factors and metalloproteinases were detected in the serum of each group by ELISA. The protein and messenger RNA (mRNA) expressions of important elements related to Toll-like receptors (TLRs)-mediated innate immune response in the synovial tissue were detected by Western blot and quantitative PCR (qPCR). The Lequesne-MG index score of the rabbits gradually increased with the modeling prolonged but decreased significantly after EA treatment, indicating that EA has a better effect on alleviating the pain and improving the dysfunction. The morphological analysis showed that the inflammation of and the damage to the synovial membrane and the cartilage tissue gradually deteriorated with the modeling prolonged. However, the synovial membrane inflammation was significantly relieved after EA treatment, and the cartilage injury showed signs of repair. The ELISA analysis showed that, with the modeling prolonged, the serum-related inflammatory factors and mechanism of metalloproteinases gradually increased but decreased after EA treatment. The tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), and matrix metalloproteinase3 (MMP3) of EA1 group were significantly lower than those of EA2 group. Both Western blot and qPCR results showed that the protein and mRNA expressions of the elements related to the innate immune response in the synovial membrane increased gradually with the modeling prolonged, but decreased significantly after EA treatment. Additionally, the expression of some components in EA1 group was significantly lower than that in EA2 group. These results confirm that synovial inflammation gradually aggravated with time from the early to mid-stage of KOA. EA alleviated the inflammation and histological changes in KOA rabbits by inhibiting the TLRs-mediated innate synovial immune response. This suggests that using EA in the early stage of KOA may achieve a desirable efficacy.

Cinnamic Aldehyde Inhibits Lipopolysaccharide-Induced Chondrocyte Inflammation and Reduces Cartilage Degeneration by Blocking the Nuclear Factor-Kappa B Signaling Pathway.

Osteoarthritis (OA), as one of the top 10 causes of physical disability, is characterized by inflammation of the synovial membrane and progressive destruction of the articular cartilage. Cinnamic aldehyde (CA), an α,ß-unsaturated aldehyde extracted from the traditional Chinese herbal medicine cinnamon (Cinnamomum verum J.Presl), has been reported to have anti-inflammatory, antioxidant, and anticancer properties. However, the anti-inflammatory effect of CA on OA remains unclear. The purpose of the present study was to investigate the effects of CA on inflammation, and cartilage degeneration in OA. A CCK-8 assay was performed to assess the potential toxicity of CA on cultured human OA chondrocytes. Following treatment with lipopolysaccharide (LPS) and CA, the expression of proinflammatory cytokines, including interleukin (IL)-1ß, IL-6, and tumor necrosis factor-alfa (TNF-α), was evaluated using quantitative real-time polymerase chain reaction (RT-qPCR) analysis, enzyme-linked immunosorbent assay, and Western blotting (WB). The production of matrix metalloproteinase-13 (MMP-13) and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS-5) was also examined using RT-qPCR and WB. Furthermore, to investigate the potential anti-inflammatory mechanism of CA, biomarkers of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway (p65, IKB-α) were detected using WB. The results demonstrated that CA significantly inhibited the expressions of IL-1ß, IL-6, TNF-α, MMP-13, and ADAMTS-5 in LPS-induced OA chondrocytes. CA dramatically suppressed LPS-stimulated NF-κB activation. Collectively, these results suggest that CA treatment may effectively prevent OA.

[Effect of Tongluo Zhitong (, TLZT) gel preparation on p53 /miR-502-5p /NF-κBp65 in synovial tissue of knee osteoarthritis].

OBJECTIVE: To observe effects of Tongluo Zhitong (, TLZT) gel preparation on p53, miR-502-5p, NF-κBp65 in synovial tissue of knee osteoarthritis (KOA), and to explore mechanism of TLZT gel preparation in treating KOA. METHODS: Thirthy-six Wistar rats aged 8 weeks and weighed 200 to 220 g (meaned 208 g) were randomly divided into normal group, model group and traditional Chinese medicine (TCM) group, 12 rats in each group. KOA model was established by modified Hulth method. After 4 weeks of modeling, TCM group treated with TLZT gel preparation for external use, 3 times daily for 2 weeks;normal group and model group were fed normally without intervention. After treatment, morphological changes of specimens in each group were observed, changes of miR-502-5p in synovial tissue were detected by qPCR, and contents of p53, NF-κBp65, IL-1ß, TNF-α, MMP-13 in synovial tissue were detected by qPCR and Western Blot respectively. RESULTS: (1)Morphological observation of specimens showed that the articular cartilage in model group was hyaline and uneven, the synovial membranes were hypertrophic and proliferative with a large number of inflammatory cells infiltrating, the joint fluid was thicker in texture;the articular cartilage in TCM group was more transparent and smooth, synovial hyperplasia was mild with a small amount of inflammatory cell infiltration, the texture of articular fluid was clear and sparse. (2) Compared with normal group, content of miR-502-5p of synovial tissue in model and TCM group were increased, mRNA and expression of p53 decreased, expression of NF-κBp65, IL-1ß, TNF-α, MMP-13 increased. (3)Compared with model group, content of miR-502-5p in synovial tissue of TCM group decreased (P<0.05), mRNA and protein expression of p53 increased (P<0.05), mRNA and protein expression of NF-κBp65, IL-1ß, TNF-α, MMP-13 decreased (P<0.05). CONCLUSION: Expression of p53, miR-502 -5p, NF -κBp65 in synovial tissue is closely related to synovial hyperplasia and inflammatory reaction, TLZT gel preparation may reduce proliferation and inflammatory reaction of KOA synovium by regulating the expression of p53, miR- 502-5p, NF-κBp65 in synovial tissues.

Morphological and Molecular Analysis Identifies the Associated Fungus ("Xianghui") of the Medicinal White Jelly Mushroom, Tremella fuciformis, as Annulohypoxylon stygium.

White jelly mushroom, Tremella fuciformis, is a popular edible mushroom with interesting medicinal properties (e.g., immunostimulating, antidiabetic). The formation of T. fuciformis basidiomes is highly dependent on the presence of a specific host fungus, both in nature and for industrial production. This host has traditionally been indicated as "Xianghui" in China, yet which or how many fungal species Xianghui comprises is unclear, with various authorities claiming different species. At present, Annulohypoxilon archeri is generally assumed to be the main Xianghui species, but this has not yet been confirmed experimentally. The implementation of older, premolecular-based research data (i.e., morphological) with present, sequence-based data to solve the identity remains confusing and studies addressing both identification methods in combination are lacking. The unclear identity of Xianghui is a major obstacle for further understanding of the important relationship between the host(s) and T. fuciformis. In this study, we collected a wild specimen of T. fuciformis together with several nearby stroma of Xianghui, cocultivated T. fuciformis with the Xianghui isolates, and observed basidiome formation. Internal transcribed spacer (ITS) sequence analysis showed that all Xianghui spore isolates belonged to the same species and both morphological analysis of sexual stages and ITS ß-tubulin and actin gene sequences of the Xianghui specified it as Annulohypoxylon stygium. The ITS sequences of the newly identified Xianghui further closely matched those of the Xianghui strains used in the mushroom industry, showing that wild and culture spawn Xianghui in China consist of A. stygium. In contrast with previous conclusions, A. stygium, and not A. archeris, seems to be the preferred host of T. fuciformis.

Study on the relation between tissues pathologies and traditional chinese medicine syndromes in knee osteoarthritis: Medical image diagnostics by preoperative X-ray and surgical arthroscopy.

OBJECTIVE: This study aims to investigate whether integration of traditional Chinese medicine and modern medicine has advantage in achieving the improved diagnosis and treatment of knee osteoarthritis. METHODS: 90 patients with knee osteoarthritis were selected from The Department of Minimal Invasive Joint of The Third Affiliated Hospital of Beijing University of Chinese Medicine from June 2013 to June 2015. They were divided into 3 groups with 30 cases per group in accordance to the syndrome differentiation of traditional Chinese medicine. The patients underwent arthroscopic surgery, and we categorized the patients having the same characterization in each group, and those having distinct difference into the three groups. Based on the arthroscopic analysis, we performed analysis of statistical data in order to analyze the relation between knee osteoarthritis under arthroscope and traditional Chinese medicine syndromes. RESULTS: There are three syndromes according to traditional Chinese medicine that can be categorized into various different groups. The synovial proliferation can be seen mostly in the syndrome of stagnation of blood stasis. The slight damage of knee joint cartilage can be seen in the syndrome of yang deficiency and cold stagnation, the severe one in the syndrome of kidney-marrow deficiency. We found that there are different pathological expressions with the various degree of the tissues damage at the knee and we categorized the knee according to their syndrome. CONCLUSION: For knee osteoarthritis, different syndromes of traditional Chinese medicine presents different tissues pathological changes at the knee joint under arthroscopy, which will provide objective basis for the diagnosis of this medical condition.

Therapeutic effects of low-frequency phonophoresis with a Chinese herbal medicine versus sodium diclofenac for treatment of knee osteoarthritis: a double-blind, randomized, placebo-controlled clinical trial.

OBJECTIVE: To evaluate the therapeutic effects of low-frequency phonophoresis with a Chinese herbal medicine (CHM) compared with sodium diclofenac (SD) for knee osteoarthritis (KOA). METHODS: In this double-blind, randomized, placebo-controlled trial, 100 KOA patients were assigned randomly to a placebo group, a CHM group, or SD group. Low-frequency phonophoresis was used to improve the efficiency of drug delivery. Pain at rest [using a visual analog scale (VAS)], pain on movement (VAS), and range of motion (degrees) in the three groups were evaluated using the Western Ontario and McMaster Universities Osteoarthritis Index (WOMACAI) scores. Safety assessments comprised emergency adverse events, as well as laboratory tests of blood biochemistry, creatinine, blood urea nitrogen, alanine aminotransferase and aspartate aminotransferase. RESULTS: Significant improvements were found after treatment in all outcome measures except stiffness and range of motion in patients in the CHMP group and SDP group (P < 0.05). No significant differences in all outcome measures were found between the CHMP group and SDP group. CONCLUSION: CHMP and SDP can show good therapeutic effects for KOA in terms of relieving pain and improving physical function.

[Study about Traditional Chinese Medicine syndrome of adolescent neck pain].

OBJECTIVE: To preliminarily research the formular about the Traditional Chinese Medicine (TCM) syndrome of adolescent neck pain. METHODS: An observation table of adolescent neck pain syndromes was formulated,and 1 397 patients with adolescent neck pain were investigated to establish a database of adolescent neck pain. The Descriptive Statistical Analysis and Hierarchical Cluster Analysis were performed by statistical software. RESULTS: Totally 60 TCM symptoms was clustered into 4 TCM syndromes by Hierarchical Cluster Analysis. The expert panel of TCM syndromes preliminarily formulate 4 TCM syndromes of adolescent neck pain by analyzing the result of Cluster Analysis and discussing their clinical experience. CONCLUSION: Adolescent neck pain is a category of Tendon Trauma's Bi-syndrome of TCM. Ying, Wei, Qi and blood block caused by exopathy, strains, and internal injury is considered as the main pathogenesis of adolescent neck pain. Base on statistical result and expert's opinions, 4 TCM syndromes about adolescent neck pain were formulated: cold-dampness syndrome, dampness-heat blockage syndrome, liver-stagnation and spleen-deficiency syndrome, Qi and Yin deficiency of both heart and kidney syndrome.

[Case-control study on therapeutic effects between bone-setting and herbal fumigation for the treatment of the obsolete malleolus joint sprains].

OBJECTIVE: To compare the clinical effects of bone-setting technique and herbal fumigation for the treatment of the obsolete malleolus joint sprains. METHODS: From March 2008 to May 2011, 76 patients were divided into treatment group (39 cases) and control group (37 cases). In the treatment group: 15 males and 24 females; the age ranged from 20 to 59 years with an average of (42.97 +/- 9.21) years; the course of disease ranged from 1 to 60 months; the average score of ankle joint function was (71.27 +/- 4.50). In the control group: 11 males and 26 females; the age ranged from 25 to 57 years with an average of (41.29 +/- 8.77) years; the course of disease ranged from 1 to 36 months with an average of (8.47 +/- 7.37) months; the average score of ankle joint function was (71.45 +/- 4.61). The patients in the treatment group were treated with bone-setting technique two times a week, and the patients in the control group were treated with herbal fumigation once a day. The ankle joint function scores and treatment effects of the two groups were compared after 3 weeks by using Baird-Jackson ankle function score. RESULTS: After 3 weeks of the treatment, the average score of ankle joint function of the treatment group was (93.44 +/- 4.91), and in the control group was (85.8 +/- 16.57), the difference has statistical significance. The treatment group score was better than that of the control group. Before and after treatment,the average ankle score of the treatment group was (71.27 +/- 4.50) and (93.44 +/- 4.91), the difference has statistical significance. Before and after treatment, the average ankle score of the control group was (71.45 +/- 4.61) and (85.81 +/- 6.57), the difference has statistical significance. In the treatment group, 16 cases got an excellent result, 18 good, 3 fair, 2 poor; in the control group, 9 cases got an excellent result, 14 good, 5 fair,9 poor. The difference has statistical significance. CONCLUSION: The bone-setting techniques and herbal fumigation treatment of obsolete malleolus joint sprains both have a certain effect, and the former is better than the latter.