Characterizing the skeletal muscle immune microenvironment for sarcopenia: insights from transcriptome analysis and histological validation.

Background: Sarcopenia is a condition characterized by the age-related loss of skeletal muscle mass and function. The pathogenesis of the disease is influenced by chronic low-grade inflammation. However, the specific changes in the immune landscape changes of sarcopenic muscle are not yet fully understood. Methods: To gain insights into the immune cell composition and interactions, we combined single-nucleus RNA sequencing data, bulk RNA sequencing dataset, and comprehensive bioinformatic analyses on the skeletal muscle samples from young, aged, and sarcopenic individuals. Histological staining was then performed on skeletal muscles to validate the distribution of immune cells in clinical samples. Results: We analyzed the transcriptomes of 101,862 single nuclei, revealing a total of 10 major cell types and 6 subclusters of immune cell types within the human skeletal muscle tissues. Notable variations were identified in the immune microenvironment between young and aged skeletal muscle. Among the immune cells from skeletal muscle microenvironment, macrophages constituted the largest fraction. A specific marker gene LYVE1 for skeletal muscle resident macrophages was further identified. Cellular subclasses included four distinct groups of resident macrophages, which play different roles in physiological or non-physiological conditions. Utilizing bulk RNA sequencing data, we observed a significant enrichment of macrophage-rich inflammation in sarcopenia. Conclusions: Our findings demonstrate age-related changes in the composition and cross-talk of immune cells in human skeletal muscle microenvironment, which contribute to chronic inflammation in aged or sarcopenia muscle. Furthermore, macrophages emerge as a potential therapeutic target, thus advancing our understanding of the pathogenesis of sarcopenia.

Identification of key mitochondria-related genes and their relevance to the immune system linking Parkinson's disease and primary Sjögren's syndrome through integrated bioinformatics analyses.

BACKGROUND: Mitochondria are the metabolic hubs of cells, regulating energy production and antigen presentation, which are essential for activation, proliferation, and function of immune cells. Recent evidence indicates that mitochondrial antigen presentation may have an impact on diseases such as Parkinson's disease (PD) and autoimmune diseases. However, there is limited knowledge about the mechanisms that regulate the presentation of mitochondrial antigens in these diseases. METHODS: In the current study, RNA sequencing was performed on labial minor salivary gland (LSG) from 25 patients with primary Sjögren's syndrome (pSS) and 14 non-pSS aged controls. Additionally, we obtained gene expression omnibus datasets associated with PD patients from NCBI Gene Expression Omnibus (GEO) databases. Single-sample Gene Set Enrichment Analysis (ssGSEA), ESTIMATE and Spearman correlations were conducted to explore the association between mitochondrial related genes and the immune system. Furthermore, we applied weighted Gene Co-expression Network Analysis (WGCNA) to identify hub mitochondria-related genes and investigate the correlated networks in both diseases. Single cell transcriptome analysis, immunohistochemical (IHC) staining and quantitative real-time PCR (qRT-PCR) were used to verify the activation of the hub mitochondria-related pathway. Pearson correlations and the CIBERSORT algorithms were employed to further reveal the correlation between hub mitochondria-related pathways and immune infiltration. RESULTS: The transcriptome analysis revealed the presence of overlapping mitochondria-related genes and mitochondrial DNA damage in patients with pSS and PD. Reactive oxygen species (ROS), the senescence marker p53, and the inflammatory markers CD45 and Bcl-2 were found to be regionally distributed in LSGs of pSS patients. WGCNA analysis identified the STING pathway as the central mitochondria-related pathway closely associated with the immune system. Single cell analysis, IHC staining, and qRT-PCR confirmed the activation of the STING pathway. Subsequent, bioinformatic analysis revealed the proportion of infiltrating immune cells in the STING-high and STING-low groups of pSS and PD. Furthermore, the study demonstrated the association of the STING pathway with innate and adaptive immune cells, as well as functional cells, in the immune microenvironment of PD and pSS. CONCLUSION: Our study uncovered a central pathway that connects mitochondrial dysfunction and the immune microenvironment in PD and pSS, potentially offering valuable insights into therapeutic targets for these conditions.

Improved Performance of QDSSCs Can Be Achieved by Constructing a Transparent Anatase TiO2@MWCNT Photoanode Based on the Bionic Mountain Lotus.

In this research, the structural characteristics of the mountain holly leaf were emulated. It was observed that after the initially uneven surface of the petals is filled with infiltrated water, it exhibits a distinctive transparent beauty after rainfall. Furthermore, the presence of leaf veins enhances the structural strength of the petals and facilitates nutrient transport. Inspired by previous studies on double-layer spin-coated films, we further developed and designed the TA TiO2@MWCNT photocathode thin film. This innovative film incorporates multiwalled carbon nanotubes (MWCNT) into a previously established TA TiO2 photocathode thin film. The inclusion of MWCNT results in the formation of a three-dimensional highway structure, where MWCNT intertwines within the TA TiO2 film. Under the operational state of immersion in the electrolyte, it maintains a level of transparency similar to that of the TA TiO2 photoanode thin film. The high-temperature sintering process results in the oxidation and depletion of MWCNTs on the surface of the film, leaving behind uniformly dispersed concave defects, thereby greatly enhancing the specific surface area. The findings demonstrate that the optoelectrode of high transparency and high specific surface area, TA TiO2@MWCNT, comprehensively enhances the performance of the solar cells. The transparent QDSSC surpasses its counterparts for the first time, achieving a power conversion efficiency (PCE) of 6.335%. This sets the stage for new materials and innovative approaches in the field of solar cells and other titanium dioxide film-related areas.

Comparative efficacy and safety of acupuncture and Western medicine for poststroke thalamic pain.

Poststroke thalamic pain (PSTP) is one of the most common sequelae following stroke. Analgesics, antidepressants, anticonvulsants, and surgical treatment are conventional treatment methods of PSTP, but these methods have limited efficacy, cost more, and cause a likelihood of adverse reactions. Clinical studies have shown that acupuncture has a significant analgesic effect on PSTP without obvious side effects. But, there is a lack of high-quality evidence concerning its effectiveness and safety to support its use. Therefore, this study aimed to evaluate the clinical efficacy and safety of acupuncture versus Western medicine for the treatment of PSTP to provide evidence to support clinical PSTP treatment. Searches were conducted to identify randomized controlled trials investigating the use of acupuncture for PSTP across six databases, including PubMed, the Cochrane Library, EMBASE, the China National Knowledge Infrastructure, Wan Fang Database, and the Chinese Scientific Journal Database VIP. RevMan 5.3 software was used for the meta-analysis. The results showed that compared with Western medicine, acupuncture had a higher total effective rate for the treatment of PSTP, reduced visual analog scale scores, increased beta-endorphin content, and decreased incidence of adverse reactions. However, the sample sizes of the included studies were insufficient, and the quality of the articles was relatively poor. In future studies, the clinical study design should be standardized and the sample size should be expanded to validate these results.

Chondrogenic potential of manganese-loaded composite scaffold combined with chondrocytes for articular cartilage defect.

Cartilage is an alymphatic, avascular and non-innervated tissue. Lack of potential regenerative capacity to reconstruct chondral defect has accelerated investigation and development of new strategy for cartilage repair. We prepared a manganese ion-incorporated natupolymer-based scaffold with chitosan-gelatin by freeze-drying procedure. The scaffold was characterized by Fourier transform infrared spectroscopy, thermogravimetric analysis, scanning electron microscopy, energy dispersive spectroscopy, compressive testing, and analysis of porosity and flexibility. Live/dead assay confirmed the good cytocompatibility of prepared scaffold on rat articular chondrocytes after 10 days and 4 weeks of culture. The manganese-loaded composite scaffold upregulated the expression of chondrogenic-related markers (Sox9, integrin, and Col II) in chondrocytes. Western blot analysis of proteins extracted from chondrocytes grown on scaffolds indicated the signaling pathways of p-Akt and p-ERK1/2 played a key role. Histological analysis following implantation of current composite scaffold loaded with chondrocytes into a rat articular cartilage defect model showed that the scaffolds promoted the formation of collagen II and cartilage repair. These findings suggested the potential of manganese-loaded scaffold to promote new cartilage formation and a promising strategy for articular cartilage engineering application.

Electronic Localization Derived Excellent Stability of Li Metal Anode with Ultrathin Alloy.

Lithium metal is an ideal anode for next-generation high-energy-density batteries. However, lithium dendrite growth has impeded its commercial application. Herein, fabricating Li-based ultrathin alloys with electronic localization and high surface work function via depositing Bi, Al, or Au metals on the surface of copper foil for in situ alloying with lithium is proposed. It is discovered that the electronic localization can induce self-smoothing effect of Li ions, as a result, significantly suppressing the growth of dendritic lithium. Meanwhile, the high surface work function can effectively alleviate side reactions between the electrolyte and lithium. With the as-obtained ultrathin alloys as anodes, excellent cycling performance is achieved. The half cells run stably after more than 120 cycles under high capacity of 4 mAh cm-2 . The S||Bi/Cu-Li full cell delivers a specific capacity of 736 mAh g-1 after 200 cycles. This work provides a new strategy for fabricating long-life and high-capacity lithium batteries.

CircRAD18 Accelerates the Progression of Acute Myeloid Leukemia by Modulation of miR-206/PRKACB Axis.

BACKGROUND: Circular RNAs (circRNAs) play a crucial role in tumorigenesis. However, the effects of circRNAs on acute myeloid leukemia (AML) remain largely unexplored. We explored the function of circRAD18 in AML development. METHODS: QRT-PCR was performed for the levels of circRAD18, RAD18, microRNA-206 (miR-206) and protein kinase CAMP-activated catalytic subunit beta (PRKACB). Cell Counting Kit-8 (CCK-8) assay and colony formation assay were utilized for cell proliferation. Flow cytometry analysis was carried out to analyze cell apoptosis and cell cycle process. Transwell assay was manipulated for cell migration and invasion. Western blot assay was conducted for protein levels. Dual-luciferase reporter assay was adopted to verify the interaction between miR-206 and circRAD18 or PRKACB. RESULTS: CircRAD18 level was increased in AML patients' blood specimens and AML cell lines compared to normal controls. CircRAD18 knockdown impeded the proliferation, migration and invasion and facilitated the apoptosis and cell cycle arrest in AML cells. Moreover, circRAD18 was identified as a sponge for miR-206, and circRAD18 knockdown-mediated effect on AML cell progression was reversed by miR-206 suppression. Additionally, PRKACB was the target gene of miR-206. MiR-206 overexpression suppressed the malignant behaviors of AML cells, while PRKACB elevation restored the effects. CONCLUSION: CircRAD18 aggravated the malignancy of AML cells through reducing miR-206 expression and elevating PRKACB expression, indicating circRAD18 might be a therapeutic target for AML.

Effects of extracorporeal shock wave therapy in patients with knee osteoarthritis: A cohort study protocol.

BACKGROUND: Osteoarthritis is the most common form of arthritis, and is a major cause of disability and chronic pain in adults. However, there is very limited evidence in the scientific literature to support the effectiveness of extracorporeal shockwave therapy (ESWT) in human knee osteoarthritis. This retrospective study aimed to compare the efficacy of ESWT treatment with sham-ESWT on pain, walking speed, physical function, and adverse effects in knee osteoarthritis. METHODS: This study will be performed and reported in accordance with the Strengthening the Reporting of Observational studies in Epidemiology checklist. We reviewed patients diagnosed with knee osteoarthritis at our academic center from 2016 to 2017. This retrospective cohort study was approved by the institutional review board in Ruijin Hospital. The primary outcome measure was pain on movement measured by a 100-cm visual analog scale. The secondary outcome measures included the Western Ontario and McMaster University Osteoarthritis Index, range of motion, and adverse effects. Statistical analysis was performed using Statistical Package for Social Sciences version 20.0 (IBM Corporation, Armonk, NY). A P-value of <.05 was defined as statistical significance. RESULTS: The hypothesis was that ESWT would be an effective treatment for improving pain and physical function in knee osteoarthritis to control symptoms. TRIAL REGISTRATION: This study protocol was registered in Research Registry (researchregistry5801).

The Inhibition of miR-873 Provides Therapeutic Benefit in a Lipopolysaccharide-Induced Neuroinflammatory Model of Parkinson's Disease.

Background and Purpose. Alterations in cholesterol homeostasis have been reported in cell and animal models of Parkinson's disease (PD), although there are inconsistent data about the association between serum cholesterol levels and risk of PD. Here, we investigated the effects of miR-873 on lysosomal cholesterol homeostasis and progressive dopaminergic neuron damage in a lipopolysaccharide-(LPS) induced model of PD. Experimental Approach. To evaluate the therapeutic benefit of the miR-873 sponge, rats were injected with a LV-miR-873 sponge or the control vector 3 days before the right-unilateral injection of LPS into the substantia nigra (SN) pars compacta, or 8 and 16 days after LPS injection. Normal SH-SY5Y cells or SH-SY5Y cells overexpressing α-synuclein were used to evaluate the distribution of α-synuclein and cholesterol in lysosomes and to assess the autophagic flux after miR-873 transfection or ABCA1 silencing. The inhibition of miR-873 significantly ameliorated the LPS-induced accumulation of α-synuclein and loss of dopaminergic neurons in the SN at the early stage. miR-873 mediated the inhibition of ABCA1 by LPS. miR-873 transfection or ABCA1 silencing increased the lysosomal cholesterol and α-synuclein levels, and decreased the autophagic flux. The knockdown of ABCA1 or A20, which are the downstream target genes of miR-873, exacerbated the damage to LPS-induced dopaminergic neurons. Conclusion and Implications. The results suggest that the inhibition of miR-873 may play a dual protective role by improving intracellular cholesterol homeostasis and neuroinflammation in PD. The therapeutic effects of the miR-873 sponge in PD may be due to the upregulation of ABCA1 and A20.

Glutamate affects the CYP1B1- and CYP2U1-mediated hydroxylation of arachidonic acid metabolism via astrocytic mGlu5 receptor.

The extrahepatic CYP enzymes, CYP1B1 and CYP2U1, have been predominantly found in both astrocytes and brain microvessels. We investigated the alteration in the production of hydroxyeicosatetraenoic acids (HETEs) from arachidonic acid (AA) mainly via CYP1B1 and CYP2U1 by glutamate. CYP1B1 and CYP2U1 mRNA levels were dose-dependently induced by glutamate in human U251 glioma cells and hCMEC/D3 blood-brain barrier cells. The increases in the CYP1B1 and CYP2U1 mRNA levels and the binding of CREB to CYP1B1 and CYP2U1 promoters following glutamate treatment were attenuated by mGlu5 receptor antagonist. The mRNA levels of CYP1B1 and CYP2U1 were increased in the cortex, hippocampus, and cerebellum from adult rats that received a subcutaneous injection of monosodium l-glutamate at 1, 3, 5, and 7 days of age; meanwhile, the protein levels of CYP1B1 and CYP2U1 in the astrocytes were induced by glutamate. Glutamate treatment significantly increased the production of 5-HETE, 8-HETE, 11-HETE, and 20-HETE in the cortex and cerebellum. These data suggested that the neuron-astrocyte reciprocal signaling can change the CYP-mediated AA metabolism (e.g. EETs and HETEs) in astrocytes via its specific receptor.

The Protective Role of Brain CYP2J in Parkinson's Disease Models.

CYP2J proteins are present in the neural cells of human and rodent brain regions. The aim of this study was to investigate the role of brain CYP2J in Parkinson's disease. Rats received right unilateral injection with lipopolysaccharide (LPS) or 6-hydroxydopamine (6-OHDA) in the substantia nigra following transfection with or without the CYP2J3 expression vector. Compared with LPS-treated rats, CYP2J3 transfection significantly decreased apomorphine-induced rotation by 57.3% at day 12 and 47.0% at day 21 after LPS treatment; moreover, CYP2J3 transfection attenuated the accumulation of α-synuclein. Compared with the 6-OHDA group, the number of rotations by rats transfected with CYP2J3 decreased by 59.6% at day 12 and 43.5% at day 21 after 6-OHDA treatment. The loss of dopaminergic neurons and the inhibition of the antioxidative system induced by LPS or 6-OHDA were attenuated following CYP2J3 transfection. The TLR4-MyD88 signaling pathway was involved in the downregulation of brain CYP2J induced by LPS, and CYP2J transfection upregulated the expression of Nrf2 via the inhibition of miR-340 in U251 cells. The data suggest that increased levels of CYP2J in the brain can delay the pathological progression of PD initiated by inflammation or neurotoxins. The alteration of the metabolism of the endogenous substrates (e.g., AA) could affect the risk of neurodegenerative disease.

The Involvement of PPARs in the Selective Regulation of Brain CYP2D by Growth Hormone.

Brain CYP2D is responsible for the synthesis of endogenous neurotransmitters such as dopamine and serotonin. This study is to investigate the effects of cerebral CYP2D on mouse behavior and the mechanism whereby growth hormone regulates brain CYP2D. The inhibition of cerebellar CYP2D significantly affected the spatial learning and exploratory behavior of mice. CYP2D expression was lower in the brain in GHR-/- mice than that in WT mice; however, hepatic CYP2D levels were similar. Brain PPARα expression in male GHR-/- mice were markedly higher than those in WT mice, while brain PPARγ levels were decreased or unchanged in different regions. However, both hepatic PPARα and PPARγ in male GHR-/- mice were markedly higher than those in WT mice. Pulsatile GH decreased the PPARα mRNA level and increased the mRNA levels of CYP2D6 and PPARγ in SH-SY5Y cells. A luciferase assay showed that PPARγ activated the CYP2D6 gene promoter while PPARα inhibited its function. Pulsatile GH decreased the binding of PPARα to the CYP2D6 promoter by 40% and promoted the binding of PPARγ to the CYP2D6 promoter by approximately 60%. The male GH secretory pattern altered PPAR expression and the binding of PPARs to the CYP2D promoter, leading to the elevation of brain CYP2D in a tissue-specific manner. Growth hormone may alter the learning and memory functions in patients receiving GH replacement therapy via brain CYP2D.

Glutamate affects the production of epoxyeicosanoids within the brain: The up-regulation of brain CYP2J through the MAPK-CREB signaling pathway.

Glutamate is the major excitatory neurotransmitter in the brain, and chronic glutamate excitotoxicity has been thought to be involved in numerous neurodegenerative diseases. We investigated the effects of glutamate at concentrations lower than the usual extrasynaptic concentrations on the production of epoxyeicosanoids mediated by brain CYP2J. Glutamate increased CYP2J2 mRNA levels in astrocytes in a dose-dependent manner, while an antagonist of the metabotropic glutamate receptor subtype 5 (mGlu5 receptor) attenuated the glutamate-induced increases in CYP2J2 levels by glutamate. Glutamate increased the binding of cAMP response element-binding protein (CREB) with the CYP2J2 promoter, and the inhibition of the MAPK signaling pathway (ERK1/2, p38, and JNK) decreased the binding of CREB with the CYP2J2 promoter following the glutamate treatment. CREB activated the CYP2J2 promoter located at -1522 to -1317bp, and CREB overexpression significantly increased CYP2J2 mRNA levels. The CYP2J2 and mGlu5 mRNA levels were higher in the frontal cortex, hippocampus, cerebellum, and brainstem in adult rats that received a subcutaneous injection of monosodium l-glutamate at 1, 3, 5, and 7days of age. The data from the partial least-squares-discriminant analysis showed the epoxyeicosanoid profile of the hippocampus from the cerebellum, brain stem, and frontal cortex. The sum of the epoxyeicosatrienoic acids (EETs) and dihydroxyeicosatrienoic acids (DHETs) was increased by 1.16-fold, 1.18-fold, and 1.19-fold in the frontal cortex, cerebellum, and brain stem, respectively, in rats treated with monosodium l-glutamate compared with the control group. The results suggest that brain CYP2J levels and CYP2J-mediated epoxyeicosanoid production can be regulated by extrasynaptic glutamate. The glutamate receptors expressed in astrocytes may mediate the regulation of drug-metabolizing enzymes and the metabolome of endogenous substances by glutamate.

Purification, characterization and in vitro antioxidant activities of polysaccharide from Chaenomeles speciosa.

A water-soluble polysaccharide named as CSP-2 was isolated and purified from Chaenomeles speciosa by DEAE-Sepharose and Sephadex G-100 column chromatography. CSP-2 was composed of galactose (Gal), rhamnose (Rha), glucose (Glc) and xylose (Xyl) with a relative molar ratio of 3.8:1.6:1.2:0.4. The average molecular weight of CSP-2 was estimated to be about 4.6×104Da. The in vitro antioxidant assay showed that CSP-2 exhibited remarkable antioxidant activities to scavenge the DPPH radical, superoxide radical, hydroxyl radical and ABTS radical and significant cytoprotective effects on H2O2-induced PC12 cells injury in a concentration-dependent manner. Overall, CSP-2 may be useful as a naturally potential antioxidant agent for application in food and medicinal fields.

Antitumor and immunomodulatory activities of a water-soluble polysaccharide from Chaenomeles speciosa.

In this study, a water-soluble polysaccharide (CSP) was successfully purified from Chaenomeles speciosa by DEAE-Sepharose and Sephadex G-100 column chromatography. CSP had a weight-average molecular weight of about 6.3 × 10(4)Da and was composed of glucose (Glc), galactose (Gal), rhamnose (Rha) and arabinose (Ara) with a relative molar ratio of 4.6:1.3:0.8:0.5. CSP could not only inhibit the growth of S180 tumor transplanted in mice, but also increase the relative spleen index and body weight of tumor bearing mice. Moreover, concanavalin A (ConA) and lipopolysaccharide (LPS) induced splenocyte proliferation and peritoneal macrophage phagocytosis were also enhanced after CSP administration. Furthermore, CSP treatment could improve delayed type hypersensitivity (DTH) and promote the secretion of IL-2, TNF-α and IFN-γ in serum. The overall findings suggest that the antitumor effect of CSP is might be associated with its potent immunostimulatory activity.

Characterization and antitumor activities of a water-soluble polysaccharide from Ampelopsis megalophylla.

A water-soluble polysaccharide named as AMP was isolated and purified from the leaves of Ampelopsis megalophylla by DEAE-52 Cellulose and Sephadex G-100 column chromatography. AMP had an average molecular weight of about 8.4 × 10(4)Da and was composed of galactose (Gal), mannose (Man), glucose (Glc), arabinose (Ara), and rhamnose (Rha) in a molar ratio of 2.7:1.6:1.1:0.6:0.3. After 10 days of AMP (50, 100, and 200mg/kg) treatment once daily in tumor-bearing mice, AMP oral administration could inhibit the growth of transplantable Sarcoma 180 (S180) tumor in mice and increase the spleen index and body weight. Furthermore, AMP also promote splenocytes' proliferation induced by concanavalin A (ConA) and lipopolysaccharide (LPS), strengthen peritoneal macrophages to devour neutral red and increase the production of interleukin-2 (IL-2), tumor necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ) in serum. These results suggest that AMP had clear antitumor activity, which might be related to its regulation of immune function in mice.

Angelica Sinensis polysaccharides stimulated UDP-sugar synthase genes through promoting gene expression of IGF-1 and IGF1R in chondrocytes: promoting anti-osteoarthritic activity.

BACKGROUND: Osteoarthritis (OA) is a chronic joints disease characterized by progressive degeneration of articular cartilage due to the loss of cartilage matrix. Previously, we found, for the first time, that an acidic glycan from Angelica Sinensis Polysaccharides (APSs), namely the APS-3c, could protect rat cartilage from OA due to promoting glycosaminoglycan (GAG) synthesis in chondrocytes. In the present work, we tried to further the understanding of ASP-3c's anti-OA activity. METHODOLOGY/PRINCIPAL FINDINGS: Human primary chondrocytes were treated with APS-3c or/and recombinant human interleukin 1ß (IL-1ß). It turned out that APS-3c promoted synthesis of UDP-xylose and GAG, as well as the gene expression of UDP-sugar synthases (USSs), insulin like growth factor 1 (IGF1) and IGF1 receptor (IGF1R), and attenuated the degenerative phenotypes, suppressed biosynthesis of UDP-sugars and GAG, and inhibited the gene expression of USSs, IGF1 and IGF1R induced by IL-1ß. Then, we induced a rat OA model with papain, and found that APS-3c also stimulated GAG synthesis and gene expression of USSs, IGF1 and IGF1R in vivo. Additionally, recombinant human IGF1 and IGF1R inhibitor NP-AEW541 were applied to figure out the correlation between stimulated gene expression of USSs, IGF1 and IGF1R induced by APS-3c. It tuned out that the promoted GAG synthesis and USSs gene expression induced by APS-3c was mediated by the stimulated IGF1 and IGF1R gene expression, but not through directly activation of IGF1R signaling pathway. CONCLUSIONS/SIGNIFICANCES: We demonstrated for the first time that APS-3c presented anti-OA activity through stimulating IGF-1 and IGF1R gene expression, but not directly activating the IGF1R signaling pathway, which consequently promoted UDP-sugars and GAG synthesis due to up-regulating gene expression of USSs. Our findings presented a better understanding of APS-3c's anti-OA activity and suggested that APS-3c could potentially be a novel therapeutic agent for OA.

Chemical composition, antimicrobial and antioxidant activities of essential oil from Ampelopsis megalophylla.

Chemical composition, antimicrobial and antioxidant activities of the essential oil of Ampelopsis megalophylla were evaluated in this research. GC-MS analysis of the essential oil revealed 42 compounds, representing 88.54% of the oil. The major compounds were borneol (10.81%), α-pinene (6.74%) and ß-elemene (6.23%). The antimicrobial activity of the essential oil was evaluated against 13 micro-organisms using the disc diffusion and broth microdilution methods. Results demonstrated higher effects of this oil against Gram-positive bacteria than the other reference strains tested. The antioxidant effect of the essential oil was evaluated by using 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and 2,20-azinobis-3-ethylbenzthiazoline-6-sulfonate scavenging assays. The essential oil exhibited moderate antioxidant activity.

Increased expression of CYP4Z1 promotes tumor angiogenesis and growth in human breast cancer.

Cytochrome P450 (CYP) 4Z1, a novel CYP4 family member, is over-expressed in human mammary carcinoma and associated with high-grade tumors and poor prognosis. However, the precise role of CYP4Z1 in tumor progression is unknown. Here, we demonstrate that CYP4Z1 overexpression promotes tumor angiogenesis and growth in breast cancer. Stable expression of CYP4Z1 in T47D and BT-474 human breast cancer cells significantly increased mRNA expression and production of vascular endothelial growth factor (VEGF)-A, and decreased mRNA levels and secretion of tissue inhibitor of metalloproteinase-2 (TIMP-2), without affecting cell proliferation and anchorage-independent cell growth in vitro. Notably, the conditioned medium from CYP4Z1-expressing cells enhanced proliferation, migration and tube formation of human umbilical vein endothelial cells, and promoted angiogenesis in the zebrafish embryo and chorioallantoic membrane of the chick embryo. In addition, there were lower levels of myristic acid and lauric acid, and higher contents of 20-hydroxyeicosatetraenoic acid (20-HETE) in CYP4Z1-expressing T47D cells compared with vector control. CYP4Z1 overexpression significantly increased tumor weight and microvessel density by 2.6-fold and 1.9-fold in human tumor xenograft models, respectively. Moreover, CYP4Z1 transfection increased the phosphorylation of ERK1/2 and PI3K/Akt, while PI3K or ERK inhibitors and siRNA silencing reversed CYP4Z1-mediated changes in VEGF-A and TIMP-2 expression. Conversely, HET0016, an inhibitor of the CYP4 family, potently inhibited the tumor-induced angiogenesis with associated changes in the intracellular levels of myristic acid, lauric acid and 20-HETE. Collectively, these data suggest that increased CYP4Z1 expression promotes tumor angiogenesis and growth in breast cancer partly via PI3K/Akt and ERK1/2 activation.

Protective effect of 20-hydroxyeicosatetraenoic acid (20-HETE) on adriamycin-induced toxicity of human renal tubular epithelial cell (HK-2).

20-Hydroxyeicosatetraenoic acid is a cytochrome P4504A11 metabolite of arachidonic acid that plays an important role in the regulation of human renal functions. In the present study, we investigated the role of 20-hydroxyeicosatetraenoic acid on adriamycin induced toxicity in human renal tubular epithelial cells. Results showed that cell viability was decreased significantly and lactate dehydrogenase activity was increased significantly in a concentration-dependent manner when human renal tubular epithelial cells were incubated with adriamycin (10⁻7-10⁻³ mol/l) for 24h. In contrast, 20-hydroxyeicosatetraenoic acid (0.1, 1, 10, 50 µmol/l) increased cell survival and decreased lactate dehydrogenase activity concentration dependently in human renal tubular epithelial cells. When 20-hydroxyeicosatetraenoic acid (10, 50 µmol/l) was co-administered with adriamycin (10⁻³ mol/l), it significantly increased cell viability and decreased lactate dehydrogenase activity. On the other hand, N-hydroxy-N'-(4-butyl-2-methylphenyl)formamidine (HET-0016) (1 µM), a selective inhibitor of 20-hydroxyeicosatetraenoic acid synthesizing enzyme exaggerated cell viability reduction and lactate dehydrogenase activity augmentation induced by adriamycin. Adriamycin suppressed the expression of cytochrome P4504A11 gene and its protein production in human renal tubular epithelial cells. Furthermore, adriamycin was more effective than N-hydroxy-N'-(4-butyl-2-methylphenyl)formamidine at lowering the expression of cytochrome P4504A11 gene and its protein. These results suggest that 20-hydroxyeicosatetraenoic acid may protect adriamycin-induced toxicity of human renal tubular epithelial cells, meanwhile, adriamycin-induced toxicity of human renal tubular epithelial cells possibly involves inhibiting cytochrome P4504A11 expression.