Pueraria lobata antioxidant extract ameliorates non-alcoholic fatty liver by altering hepatic fat accumulation and oxidative stress.

ETHNOPHARMACOLOGICAL RELEVANCE: Pueraria lobata is essential medicinal and edible homologous plants widely cultivated in Asian countries. Therefore, P. lobata is widely used in the food, health products and pharmaceutical industries and have significant domestic and international market potential and research value. P. lobata has remarkable biological activities in protecting liver, relieving alcoholism, antioxidation, anti-tumor and anti-inflammation in clinic. However, the potential mechanism of ethyl acetate extract of Pueraria lobata after 70% alcohol extraction (APL) ameliorating nonalcoholic fatty liver disease (NAFLD) has not been clarified. AIM OF THE STUDY: This study aimed to investigate the ameliorative effect of P. lobata extract on human hepatoma cells and injury in rats, and to evaluate its therapeutic potential for ameliorating NAFLD. METHODS: Firstly, the effective part of P. lobata extract was determined as APL by measuring its total substances and antioxidant activity. And then the in vitro and in vivo models of NAFLD were adopted., HepG2 cells were incubated with palmitic acid (PA) and hydrogen peroxide (H2O2). In order to evaluate the effect of APL, Simvastatin and Vitamin C (VC) were used as positive control. Various parameters related to lipogenesis and fatty acid ß-oxidation were studied, such as intracellular lipid accumulation, reactive oxygen species (ROS), Western Blot, mitochondrial membrane potential, apoptosis, and the mechanism of APL improving NAFLD. The chemical components of APL were further determined by HPLC and UPLC-MS, and molecular docking was carried out with Keap1/Nrf2/HO-1 pathway related proteins. RESULTS: APL significantly reduced lipid accumulation and levels of oxidative stress-related factors in vitro and in vivo. Immunohistochemical、Western Blot and PCR analysis showed that the expressions of Nrf2 and HO-1 were up-regulated in APL treatment. The Nrf2 inhibitor ML385 can block the rescue by APL of cellular oxidative stress and lipid accumulation induced by H2O2 and PA, demonstrating its dependence on Nrf2. UPLC/MS analysis showed that there were 3'-hydroxyl puerarin, puerarin, 3'-methoxy puerarin, daidzein, genistin, ononin, daidzin and genistein. CONCLUSION: This study further clarified the mechanism of P. lobata extract in improving NAFLD, which provided a scientific basis for developing new drugs to protect liver injury and laid a solid foundation for developing P. lobata Chinese herbal medicine resources.

Enhancing calvarial defects repair with PDGF-BB mimetic peptide hydrogels.

Addressing bone defects represents a significant challenge to public health. Localized delivery of growth factor has emerged as promising approach for bone regeneration. However, the clinical application of Platelet-Derived Growth Factor (PDGF) is hindered by its high cost and short half-life. In this work, we introduce the application of PDGF-mimicking peptide (PMP1) hydrogels for calvarial defect restoration, showcasing their remarkable effectiveness. Through osteogenic differentiation assays and q-PCR analyses, we demonstrate PMP1's substantial capacity to enhance osteogenic differentiation of bone marrow mesenchymal stem cell (BMSC), leading to increased expression of crucial osteogenic genes. Further molecular mechanistic investigations reveal PMP1's activation of the PI3K-AKT-mTOR signaling pathway, a key element of its osteogenic effect. In vivo experiments utilizing a rat calvaria critical-sized defect model underscore the hydrogels' exceptional ability to accelerate new bone formation, thereby significantly advancing the restoration of calvaria defects. This research provides a promising bioactive material for bone tissue regeneration.

Structural properties of Kudzu protein enzymatic hydrolysate and its repair effect on HepG2 cells damaged by H2O2 oxidation.

We investigated the structural properties, foaming capacity and foaming stability, antioxidant activity, and amino acid composition of Kudzu protein (KP) and Kudzu protein hydrolysate (KPH). The peptide sequence of KPH was analyzed using ultra performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS), and the binding ability of the peptide sequence to Keap1 was predicted through molecular docking simulations. The electrophoresis and molecular weight distribution analysis results showed that the molecular weight of KPH was significantly lower than that of KP, with a mean molecular weight of approximately 2000-5000 Da. The structures and properties were characterized using Fourier transform infrared spectroscopy, relative fluorescence, and circular dichroism. The results showed that KP exposed a large number of hydrophobic groups after enzymatic hydrolysis, and its structure changed from α-helical to random coils. KPH has a higher foaming capacity (200%) and foaming stability (97.5%) than KP, which may be related to the change in structure. These results indicate that moderate hydrolysis can improve the functional properties of KP, providing a new opportunity for its application as a food ingredient. The antioxidant assay results showed that KP and KPH had a good hydroxyl radical, superoxide anion, 1,1-diphenyl-2-picrylhydrazyl (DPPH), and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) scavenging capacity and a high reducing capacity. KPH exerted better antioxidant effects than KP. The scavenging rates for DPPH, ABTS, hydroxyl radicals, and superoxide anions were 89.31%, 93.14%, 85.74%, and 58.29%, respectively, and its reducing capacity was 2.191, which may be related to the increase in amino acids with antioxidant activity after enzymolysis. In vitro, KP and KPH could significantly repair H2O2-induced oxidative damage in HepG2 cells, reduce the apoptosis rate, activate the Nrf2-Keap1 signaling pathway, reduce the accumulation of reactive oxygen species and malondialdehyde after oxidative damage, increase the activities of superoxide dismutase and glutathione (GSH) peroxidase, and increase the content of GSH and the total antioxidant capacity. Twenty-one peptide components were identified in KPH using UPLC-MS/MS, and the binding ability of 21 peptide components to Keap1 was analyzed through molecular docking technology. The results showed that all 21 peptides in KPH had good antioxidant activity, and real-time quantitative PCR (qRT-PCR) analysis was conducted to further explain the high antioxidant activity of KPH at the genetic level. These results show that KP and KPH are suitable for preparing antioxidant foods and related health foods to prevent oxidation-related diseases. KPH has more beneficial effects than KP.

Immunomodulatory activity of semen Ziziphi Spinosae protein: a potential plant protein functional food raw material.

Semen Ziziphi Spinosae protein (SZSP) is a new plant protein resource with good food functional properties and health care function. However, the biological activity of SZSP has not been further studied, which greatly limits the development and utilization of SZSP in the food industry. The aim of this study was to investigate the protective effect of SZSP on immunosuppressed mice and its inhibitory effect on immune-stimulated RAW264.7 cells. The results demonstrated that SZSP remarkably improved the immunomodulatory secretion in serum (interleukin-2, tumor necrosis factor-α [TNF-α], interferon-γ, immunoglobulin-A, immunoglobulin-G, immunoglobulin-M) and primary macrophages (nitric oxide, interleukin-1ß, TNF-α) and promoted the NK-cell killing activity of primary splenocytes in CTX-induced immunosuppression mice. Immunohistochemical analysis results indicated that the secretion of CD4+ and CD8+ in the spleen and thymus can be regulated by SZSP, leading to inhibition of the damage induced by cyclophosphamide in mice. Meanwhile, in order to clarify the immunomodulatory mechanism of SZSP, we showed that SZSP significantly inhibited the secretion of NO, interleukin-6, and TNF-α and reduced the phosphorylation expression of p-ERK, p-JNK, and p-IκBα in lipopolysaccharide-stimulated RAW264.7 cells. Therefore, the immunomodulatory effect of SZSP may be related to the activation of MAPKs and NF-κB signaling pathways. Based on the above studies, the preliminary purification of SZSP was continued, and S1F2G1 with immunomodulatory activity was obtained. Taken together, SZSP has an immunoregulatory effect in vivo and in vitro and may be a favorable candidate of functional food raw material for regulating immune responses.

Epidemiological updates of post-traumatic related limb osteomyelitis in china: a 10 years multicentre cohort study.

BACKGROUND: Post-traumatic related limb osteomyelitis (PTRLO) is a complex bone infection. Currently, there are no available microbial data on a national scale that can guide appropriate antibiotic selection, and explore the dynamic changes in dominant pathogens over time. This study aimed to conduct a comprehensive epidemiological analysis of PTRLO in China. METHODS: The study was approved by the Institutional Research Board (IRB), and 3526 PTRLO patients were identified from 212 394 traumatic limb fracture patients at 21 hospitals between 1 January 2008 and 31 December 2017. A retrospective analysis was conducted to investigate the epidemiology of PTRLO, including changes in infection rate (IR), pathogens, infection risk factors and antibiotic resistance and sensitivity. RESULTS: The IR of PTRLO increased gradually from 0.93 to 2.16% (Z=14.392, P <0.001). Monomicrobial infection (82.6%) was significantly higher than polymicrobial infection (17.4%) ( P <0.001). The IR of Gram-positive (GP) and Gram-negative (GN) pathogens showed a significant increase from the lowest 0.41% to the highest 1.15% (GP) or 1.62% (GN), respectively. However, the longitudinal trend of GP vs. GN's composition did not show any significance (Z=±1.1918, P >0.05). The most prevalent GP strains were Methicillin-sensitive Staphylococcus aureus (MSSA) (17.03%), Methicillin-resistant Staphylococcus aureus (MRSA) (10.46%), E. faecalis (5.19%) and S. epidermidis (4.87%). In contrast, the dominant strains GN strains were Pseudomonas Aeruginosa (10.92%), E. cloacae (10.34%), E. coli (9.47%), Acinetobacter Baumannii (7.92%) and Klebsiella Pneumoniae (3.33%). In general, the high-risk factors for polymicrobial infection include opened-fracture (odds ratio, 2.223), hypoproteinemia (odds ratio, 2.328), and multiple fractures (odds ratio, 1.465). It is important to note that the antibiotics resistance and sensitivity analysis of the pathogens may be influenced by complications or comorbidities. CONCLUSIONS: This study provides the latest data of PTRLO in China and offers trustworthy guidelines for clinical practice. (China Clinical Trials.gov number, ChiCTR1800017597).

Yuhong ointment ameliorates inflammatory responses and wound healing in scalded mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Yuhong ointment (YHO) is famous for its efficacy in clearing away heat and dampness, reducing swelling and relieving pain, and it has been used for more than 600 years. Scalding damages the skin's defense function, resulting in a large number of necrotic tissues and cells on the wound surface, which favors bacterial growth and inflammation. If the inflammation reaction is not controlled on time, it may lead to reduced immunity and cause complications such as infection. Yuhong ointment can promote wound healing in scalded mice, but its potential pharmacological mechanism is still unclear. AIM OF THE STUDY: This study focused on identifying the active ingredients of YHO and on investigating the performance of YHO in terms of anti-inflammatory activity and scald wound healing activity. MATERIALS AND METHODS: High-performance liquid chromatography (HPLC) and high-performance liquid chromatography-mass spectrometry (HPLC-MS) were performed to identify the active ingredients of YHO. The performance of transdermal delivery of YHO was studied via HPLC for analyzing the ingredients of the exposed skin liquid of mice. Enzyme-linked immunosorbent assay (ELISA) analysis, immunohistochemistry, and real-time fluorescence quantitative PCR (qRT-PCR) were used to investigate the anti-inflammatory and scald wound healing activity of YHO. RESULTS: A total of 41 components of YHO were identified via HPLC and HPLC-MS for the first time. In the transdermal delivery experiment, the cumulative amounts of chlorogenic acid, sesamol, ferulic acid, and L-shikonin were calculated to be 342.28, 567.89, 384.54, and 528.67 µg/cm2, respectively. Pharmacological activity experiments indicated that these four kinds of drugs exhibited different degrees of therapeutic effects on scald. Specifically, YHO high-dose (YHO-H) group showed better therapeutic ability (P < 0.01) than FN and MB group. Furthermore, the immune function of the YHO group was enhanced due to the continuous increment of the levels of Hydroxyproline (HYP), Immunoglobulin G (IgG), and vascular endothelial growth factor (VEGF) and simultaneous decrement of the levels of TNF-α, TNF-ß, IL-10, and IL-6 in the skin wound. Histological results showed that the thickening of skin tissue was alleviated after treatment with YHO. Moreover, the expression of substance P (SP), calcitonin gene related peptide (CGRP) and transient receptor potential vanilloid-1 (TRPV1) was inhibited, and the expression of VEGF was promoted by YHO (P < 0.01). The qRT-PCR test results indicated that the YHO group exhibited better inhibitory effect on interleukin 6 (IL-6), interleukin 10 (IL-10), transforming growth factor-beta (TGF-ß), and Smad-3 mRNA expression levels than the other groups. CONCLUSIONS: In this work, the active ingredients of YHO were identified via HPLC and HPLC-MS analysis. Importantly, YHO showed great advantages in transdermal delivery and scald wound healing, which can be attributed to the both anti-inflammatory and tissue regeneration mechanisms. Therefore, this work not only identified the active ingredients of YHO but also revealed the potential pharmacological mechanism of YHO for the healing of scald.

A dual-mode optical fiber sensor for SERS and fluorescence detection in liquid.

The in vivo detection of biomarkers in a liquid environment is very important for the early diagnosis of diseases. Spectroscopy methods are employed in ultraviolet-visible-infrared wavelengths, fluorescence or Raman spectra are detected for clinical diagnose. The dual-mode image can provide more diagnostic information and has been realized in some research work. However, there is still lacking simple and sensitive dual-mode sensors to satisfy the in vivo detecting demands. In this paper, a dual-mode fiber sensor for Surface-enhanced Raman Scattering (SERS) and fluorescence detection is proposed. The sensor is formed by a tapered optical fiber, half of the fiber tip surface is coated with Ag nanoparticles. In the detection of Rhodamine 6G (R6G) aqueous solution, the minimum detectable concentrations in SERS and fluorescence tests are of the same order of magnitude. By combining the Raman spectral features and the fluorescence intensity, the recognition and quantitation of target molecules were obtained reliably. It is the first time, to our knowledge, that the Raman-fluorescence dual-mode detection is realized in one single fiber, which was manufactured with micro-machinery techniques. It is a label-free, general-purpose fiber sensor, which can be applied for liquid biopsy, helping to diagnose and treat diseases in vivo.

Skin cancer signal associated with phosphodiesterase inhibitors: gaining insight through the FDA pharmacovigilance database.

BACKGROUND: Panoramic views of post-marketing safety profiles, such as cancer signal, of phosphodiesterase 5A (PDE5A) inhibitors have yet to be fully evaluated. RESEARCH DESIGN AND METHODS: Data from the FDA Adverse Event Reporting System (FAERS) concerning the timeframe between January 1st, 2004 to 30 June 2022 was analyzed through a disproportionality study to understand the association between sildenafil, tadalafil, and vardenafil and cancer. This association was identified using the Reporting odds ratio (ROR) and Bayesian Confidence Propagation Neural Network (BCPNN) approaches. RESULTS: Sildenafil associated ROR values and IC025 ranged from 9.19 (95% CI 7.72-10.94, IC025 2.77) for metastatic malignant melanoma to 132.23 (95% CI 95.49-183.11, IC025 4.69) for malignant melanoma stage II. Tadalafil associated ROR and IC025 ranged from 6.79 (95% CI 5.41-8.54, IC025 2.27) for metastatic malignant melanoma to 180.17 (95% CI 130.11-249.50, IC025 4.89) for malignant melanoma stage II. Vardenafil associated ROR and IC025 ranged from 23.38 (95% CI 15.20-35.96, IC025 2.63) for metastatic malignant melanoma to 245.77 (95% CI 154.42-391.16, IC025 2.10) for malignant melanoma stage III. CONCLUSIONS: This study supports the association between sildenafil, tadalafil, and vardenafil with skin cancer signal in erectile dysfunction (ED) patients.

Physico-chemical properties, antioxidant activity, and ACE inhibitory activity of protein hydrolysates from wild jujube seed.

Wild jujube seed protein (WJSP) as one kind of functional food material has attracted much attention due to its highly nutritive and medicinal value in anti-inflammatory and improving immunomodulatory ability. However, owing to its large molecular weight and complex structure, biological activities of WJSP were greatly limited and cannot be fully utilized by the human body. Therefore, how to improve the bioavailability of WJSP and develop promising WJSP nutritious materials is a great challenge. In this work, wild jujube seed protein hydrolysates (WJSPHs) were prepared from WJSP via enzymatic hydrolysis method, and their physico-chemical properties, antioxidant activity, and angiotensin converting enzyme (ACE) inhibitory activity in vitro have been investigated for the first time. SDS-PAGE electrophoresis and size-exclusion chromatographic results indicate that WJSPHs have lower molecular weight distribution (< 5,000 Da) than WJSP. Circular dichroism (CD) spectroscopy and Fourier transform infrared spectroscopy (FTIR) results illustrated that random coil is the main secondary structure of WJSPHs. Antioxidant experiments indicate that WJSPHs exhibit high radicals-scavenging ability of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals (94.60%), 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonate) (ABTS+ ) radicals (90.84%), superoxide radicals (44.77%), and hydroxyl radicals (47.77%). In vitro, WJSPHs can significantly decrease the accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA), and increase the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in HepG2 cells. Moreover, ACE activity was found that can be significantly inhibited by WJSPHs (73.02%). Therefore, all previously mentioned results suggest that WJSPHs may be a promising antioxidant food to prevent oxidative-related diseases in future. PRACTICAL APPLICATION: This study shows that WJSPHs exhibit high antioxidant activity and ACE inhibitory activity in vitro, which provide potential application value as antioxidant peptides to prevent oxidative-related diseases.

Association between cyclin-dependent kinase 4/6 inhibitors and venous thromboembolism: analysis of F.A.E.R.S. data.

OBJECTIVES: Real world studies have started to emerged on occurrence of venous thromboembolism (VTE) with cyclin-dependent kinase 4 and 6 (CDK4/6) inhibitors, but still deserve constant surveillance and evaluation. This study was to analyze this association. METHODS: Adverse event cases were acquired from the US Food and Drug Administration Adverse Event Reporting System (FAERS) database betweenJanuary 1st 2015 and December 31st 2020.Signals indicating association between CDK 4/6 inhibitors and VTE were identified by reporting odds ratio (ROR). RESULTS:  CDK 4/6 inhibitors had a total of 631 reports of VTE (ROR 1.44, 95% CI 1.33-1.55) compared with non-CDK 4/6 inhibitors. Palbociclib (ROR 1.42, 95% CI 1.09-1.88) demonstratedthe highest number of VTE reports, followed by ribociclib (ROR 1.41, 95% CI 1.29-1.54) and abemaciclib (ROR 0.92, 95% CI 0.72-1.17). CONCLUSIONS: Although it is not able to confirm the casual relationship between VTE and CDK4/6 inhibitors, this study suggested signal of VTE reporting in patients receiving CDK4/6 inhibitors, which is likely to reflect a potential association. The results may enhance physicians' awareness of the potential side effect of VTE associated with CDK 4/6 inhibitors. An early recognition of VTE signs/symptoms could decrease the morbidity and severity of such adverse events.

KIF26B Silencing Prevents Osseous Transdifferentiation of Progenitor/Stem Cells and Attenuates Ectopic Calcification in a Murine Model.

Ectopic calcification is an osteogenic process that leads to the formation of inappropriate bone within intra-articular soft tissues, often in response to injury or surgery. The molecular mechanisms governing this phenotype have yet to be determined. Using a population genetics approach, we identified an association of the kinesin superfamily member 26b (Kif26b) with injury-induced ectopic calcification through quantitative trait locus analysis of recombinant inbred mouse strains, consistent with a genomewide association study that identified KIF26B as a severity locus for ectopic calcification in patients with hip osteoarthritis. Despite these associations of KIF26B with ectopic calcification, its mechanistic role and functional implications have not yet been fully elucidated. Here, we aim to decipher the functional role of KIF26B in osseous and chondrogenic transdifferentiation of human and murine progenitor/stem cells and in a murine model of non-invasive injury-induced intra-articular ectopic calcification. We found that KIF26B ablation via lentivirus-mediated shRNA significantly arrested osteogenesis of progenitor/stem cells and suppressed the expression of typical osteogenic marker genes. Conversely, KIF26B loss-of-function increased chondrogenesis as demonstrated by enhanced Safranin-O staining and by the elevated expression of chondrogenic marker genes. Furthermore, cell function analysis revealed that KIF26B knockdown significantly decreased cell viability and proliferation and induced cellular apoptosis. Mechanistically, loss of osteogenesis was reverted by the addition of a Wnt agonist, SKL2001, demonstrating a role of KIF26B in canonical Wnt/ß-catenin signaling. Finally, intra-articular delivery of Kif26b shRNA in B6-129SF2/J mice significantly hampered the development of intra-articular ectopic calcification at 8 weeks after injury compared with mice treated with non-target scrambled shRNA. In summary, these observations highlight that KIF26B plays a crucial role in ectopic bone formation by repressing osteogenesis, but not chondrogenesis, potentially via modulating Wnt/ß-catenin signaling. These findings establish KIF26B as a critical determinant of the osteogenic process in pathologic endochondral bone formation and an actionable target for pharmacotherapy to mitigate ectopic calcification (and heterotopic ossification). © 2021 American Society for Bone and Mineral Research (ASBMR).

Use of a Double Reverse Traction Repositor versus a Traction Table for the Treatment of Intertrochanteric Femur Fractures: A Comparative Study.

OBJECTIVE: The aim of the present study was to compare the clinical results for unstable femoral intertrochanteric fractures treated with a double reverse traction repositor (DRTR) and those treated using a traction table with the Asia proximal femoral nail antirotation (PFNA-II). METHODS: A retrospective study was performed including 95 patients with AO/OTA type 31-A2 and 31-A3 unstable femoral intertrochanteric fractures who underwent DRTR or traction table-facilitated PFNA-II nailing from April 2015 to December 2018 in our traumatic center. Demographics, duration of operation, blood loss, part loading time after surgery, fracture healing time, and early and late complications were assessed. Clinical and radiological outcomes were collected to compare the differences between the two groups. RESULTS: A total of 95 unstable intertrochanteric fracture patients treated with the PFNA-II were analyzed. Of these cases, 56 patients were treated with a DRTR and the other 39 patients were treated using a traction table to achieve fracture reduction. No patients died during surgery and hospitalization. There were no significant differences in respect to demographics and fracture characteristics of cases enrolled. The total operative time was significantly longer in the traction table group than in the DRTR group (72.5 ± 6.1 min for the traction table and 63.0 ± 4.1 min for the DRTR group, P < 0.001). No significant differences were observed in intraoperative blood loss and duration of hospitalization. The periods of follow up ranged from 12 to 31 months among all patients. At the last follow up, the Harris hip score (HHS) in the DRTR group was excellent in 10 patients (17.9%), good in 36 (64.3%), fair in 8 (14.3%), and poor in 2 (3.6%). These scores were comparable to those in the traction table group, which were: excellent in 8 patients (20.5%), good in 24 (61.5%), fair in 6 (15.4%), and poor in 1 (2.6%). Regarding the radiological evaluation, excellent rates of reduction rate were achieved in 39 cases (69.6%) in the DRTR group, which was comparable to 19 cases (48.7%) in the traction table group. In addition, the mean fracture healing time after surgery was 20.6 ± 2.3 weeks in the DRTR group and 21.4 ± 3.4 weeks in the traction table group, which did not reach a significant difference (P = 0.18). During the follow up, 6 cases of thigh pain, 4 cases of deep vein thrombosis, and 1 case of fracture of the anterior superior iliac spine were reported in the DRTR group. In the traction table group, there were 2 cases of deep vein thrombosis and 3 cases of thigh pain. CONCLUSION: When using the PFNA-II for unstable intertrochanteric fractures, the DRTR was superior to the traction table in respect to operative time and duration of patient position, despite an additional ipsilateral anterior superior iliac spine (ASIS) incision and drilling of the ASIS and the femur condyle.

Antioxidant, Anti-Inflammatory, and Cytotoxic Properties and Chemical Compositions of Filipendula palmata (Pall.) Maxim.

Filipendula palmata (Pall.) Maxim. remains unexplored and underutilized resources with a high potential to improve human health. In this study, a new ursane-type triterpenoid, namely, 2α, 3ß-dihydroxyurs-12-en-28-aldehyde (compound 10), and other 23 known compounds were isolated. 5 triterpenoids (compounds 6, 8, and 10-12), 11 flavonoids (compounds 13-15 and 17-24), 6 phenolic compounds (compounds 1, 2, 4, 5, 9, and 16), 2 sterols (compounds 3 and 7) were isolated from the aqueous solution extract of the aerial parts of F. palmata. The structures of all compounds were elucidated by the use of extensive spectroscopic methods such as infrared spectroscopy (IR), high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), 1H-NMR, and 13C-NMR. The solvent extractions of ethyl acetate fraction were evaluated for antioxidant activities using DPPH (2, 2-diphenyl-1-picrylhydrazyl) and ABTS+ (2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) methods. The anti-inflammatory effects of the compounds were evaluated in lipopolysaccharide- (LPS-) stimulated RAW 264.7 macrophages. The extract cytotoxicity on the cancer cell lines MCF-7, HeLa, 4T1, and A549 was determined by MTT assay. As a result, compounds 10, 11, and 12 exhibited better antioxidant activity compared to the other compounds. Compounds 8-24 had different inhibitory effects on the release of NO, TNF-α, and IL-6 in LPS-stimulated RAW 264.7 cells. The new compound has shown a significant inhibiting effect on cancer cells, and the cell inhibition rate increased in a dose-dependent manner. Further research to elucidate the chemical compositions and pharmacological effects of F. palmata is of major importance towards the development and foundation of clinical application of the species.

Long non-coding RNA MEG3 inhibits chondrogenic differentiation of synovium-derived mesenchymal stem cells by epigenetically inhibiting TRIB2 via methyltransferase EZH2.

Osteoarthritis (OA) is a highly prevalent skeletal disease. Mesenchymal stem cell-derived cartilage tissue engineering is a clinical method used for OA treatment. Investigations on the molecular regulatory mechanisms of the chondrogenic differentiation of synovium-derived mesenchymal stem cells(SMSCs) will help promote its clinical applications. In this study, bioinformatics analysis from three different databases indicated that the long non-coding RNA (lncRNA) MEG3 may regulate the chondrogenic differentiation of SMSCs by targeting TRIB2. We then performed assays and found that both knockdown of MEG3 or overexpression of TRIB2 can stimulate the chondrogenic differentiation of SMSCs and increase Col2A1 and aggrecan expression. Knockdown of MEG3 can induce the expression of TRIB2; conversely, overexpression of MEG3 can inhibit the expression of TRIB2. Futhermore, knockdown of the TRIB2 can rescue the MEG3 silencing-mediated promotion of chondrogenic differentiation. Moreover, RNA immunoprecipitation(RIP) and RNA pull-down assays demonstrated that MEG3 can interact with EZH2, thus recruiting it to induce H3K27me3, which promotes the methylation of TRIB2 by binding with the promoter of TRIB2 in SMSCs. Additionally, EZH2 silencing significantly rescued the MEG3 overexpression-mediated inhibition of TRIB2 expression and chondrogenic differentiation of SMSCs. Taken together, these data indicated that MEG3 regulates chondrogenic differentiation by inhibiting TRIB2 expression through EZH2-mediated H3K27me3.

[Suppression of miR-30a/HMGA2-mediated autophagy in osteosarcoma cells impacts chemotherapeutics-induced apoptosis].

OBJECTIVE: To investigate the effect of miR-30a/HMGA2-mediated autophagy in osteosarcoma cells on apoptosis induced by chemotherapeutics. 
 Methods: A total of 30 osteosarcoma tissues of sensitive and resistant to chemotherapeutics were divided into a chemotherapy-sensitive group and a chemotherapy-resistant group. The mRNA expression levels of miR-30a and high mobility group protein A2 (HMGA2) in the chemotherapy-sensitive group and the chemotherapy-resistant group, and the mRNA expression levels of miR-30a in osteosarcoma U2-OS cells treated by cisplatin, doxorubicin and methotrexate at different concentrations were detected by real-time PCR. The expression levels of autophagy related protein Beclin 1, microtubule associated protein 1 light chain 3B (LC3B) and autophagy factor P62 were detected by Western blotting. The osteosarcoma U2-OS cells were transfected with miR-30a mimics and miR-30a inhibitors to construct a miR-30a high expression group, a miR-30a low expression group and a control group. The expression levels of Beclin 1, LC3B and P62 in osteosarcoma U2-OS cells after treatment of cisplatin and doxorubicin in these 3 groups were detected by Western blotting; the level of autophagy was detected by monodansylcada (MDC) staining; the level of ROS was detected by dihydroethidium (DHE); the level of cell surviving rate was detected by cell counting kit-8 (CCK-8); the level of apoptosis was detected by annexin APC/PI double staining; the level of mitochondria oxidative damage was detected by mitochondrial membrane potential assay kit with JC-1 (JC-1 method). The interaction between miR-30a and HMGA2 was detected by dual luciferase reporter assay. The osteosarcoma U2-OS cells were transfected with HMGA2 mimics and HMGA2-shRNA to construct a high HMGA2 group, a low HMGA2 group, and a control group. The expression levels of Beclin 1, LC3B and P62 in osteosarcoma U2-OS cells after the treatment of cisplatin were detected by Western blotting.
 Results: The level of miR-30a in the chemotherapy-resistant tissues was significantly lower than that in the chemotherapy-sensitive tissues (P<0.05), and the expression of HMGA2 was opposite comparing to that of miR-30a (P<0.05). After the treatment by low concentration (5 µmol/L) of chemotherapeutics, the level of miR-30a was down-regulated in osteosarcoma U2-OS cells, accompanied with up-regulation of Beclin 1 and LC3B (P<0.01) and down-regulation of P62 (P<0.01). Compared with the control group, the expression levels of Beclin 1 and LC3B were significantly decreased (P<0.05), and the expression level of P62 was significantly increased (P<0.05) in the miR-30a high expression group, which was opposite in the miR-30a low expression group. In the miR-30a high expression group treated by chemotherapeutics, the level of autophagy and the cell survival rate were lower than those in group with low expression of miR-30a, while the levels of ROS, the mitochondrial oxidative damage and the apoptosis were higher than those in group with low expression of miR-30a (all P<0.05). The targeting interaction between HMGA2 and miR-30a were verified by dual luciferase reporter assay. Compared with the control group, the expression levels of Beclin 1 and LC3B were significantly increased (P<0.05), and the expression level of P62 was significantly decreased (P<0.05) in the HMGA2 high expression group, which was opposite in the HMGA2 low expression group.
 Conclusion: Suppression of miR-30a/HMGA2-mediated autophagy in osteosarcoma cells is likely to enhance the therapeutic effect of chemotherapeutics.

Multiple rare causes of post-traumatic elbow stiffness in an adolescent patient: A case report and review of literature.

BACKGROUND: Joint stiffness after elbow surgery is not a rare complication, and is always accompanied by deformity. The causes of joint stiffness are multiple in different patients, and divided into intrinsic and extrinsic causes. Herein, we report an unusual case of posttraumatic elbow stiffness due to multiple and rare causes. CASE SUMMARY: A 19-year-old male was hospitalized with the loss of motion of the left elbow for over ten years. Left limb computed tomography revealed left elbow stiffness with bony block and connection. The patient underwent surgery, and the etiology of joint stiffness was found to be a rare combination of common and uncommon causes. During an 18-mo follow-up period, the patient's left elbow had normal motion and he was symptom-free. CONCLUSION: However, this case combined with multiple and rare causes highlights that the patient with scar physique is likely to be accompanied with more severe soft tissue, nerve contracture, and heterotypic ossification, even during recurrence.

Dual delivery of encapsulated BM-MSCs and BMP-2 improves osteogenic differentiation and new bone formation.

Stem cell-based therapies provide a promising approach for bone repair. In the present work, we developed a novel 3D vehicle system for dual-delivery of encapsulated bone marrow mesenchymal stem cells (BM-MSCs) and bone morphogenetic protein-2 (BMP-2) for treatment of large bone defects. The vehicle system consists of sodium alginate microcapsules and polylactic acid (PLLA) microspheres. BM-MSCs are encapsulated in the microcapsules, and BMP-2 proteins are encapsulated in the PLLA microspheres. This vehicle system acted as a multicore structure for sustained release of BMP-2, which enabled pulsed dosing induction of osteogenic differentiation of the co-embedded BM-MSCs. in vitro experiments showed that the loaded BMP-2 was constitutively released up to 30 days. Bioactivity of the incorporated BMP-2 in the microspheres was preserved and osteogenic differentiation of the BM-MSCs in the microcapsules was improved. In vivo, osteogenesis studies demonstrated that satisfactory degree of repair of a rat calvarial defect was achieved with the delivery of either encapsulated BM-MSCs alone or encapsulated BMP-2 alone. Transplantation of encapsulated both BM-MSCs and BMP-2 exhibited the greatest repair potential following 4- or 8-weeks treatment. In conclusion, microencapsulation of BM-MSCs and BMP-2 promoted the maturity of newly generated bone and improved new bone formation. Transplantation of BM-MSCs and BMP-2 in our novel 3-D vehicle system is a promising strategy for regenerative therapies of large bone defects.

Associations between blood cadmium levels and cognitive function in a cross-sectional study of US adults aged 60 years or older.

OBJECTIVES: The relationship between cadmium exposure and cognition has been well studied in children. However, the association between environmental cadmium exposure and cognitive function has not been researched extensively in older adults. Our goal was to evaluate the association between cognitive function and blood cadmium levels in US adults aged 60 years or older. DESIGN: A cross-sectional study. SETTING: The US National Health and Nutrition Examination Survey (NHANES). PARTICIPANTS: A total of 2068 adults aged 60 years or older who completed four cognitive assessment tests and blood cadmium detection in two waves of NHANES (2011-2014). MAIN OUTCOME MEASURES: Cognitive assessment was conducted by household interview or at a Mobile Examination Center (MEC) using the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) Word List Learning Test, the CERAD Word List Recall Test, the Animal Fluency Test and the Digit Symbol Substitution Test (DSST). We created a composite cognitive z-score to represent global cognitive function. RESULTS: The median blood cadmium concentration in the study participants was 0.35 µg/L, and the IQR was 0.24-0.56 µg/L. In linear regression analyses, adjusting for demographics, behaviour and medical history, blood cadmium as a continuous variable was inversely associated with the composite z-score (µg/L, ß=-0.11, 95% CI -0.20 to -0.03). Similarly, there was a significant association between quartiles of blood cadmium and composite z-score, with somewhat lower scores in the upper quartile of exposure (blood cadmium ≥0.63 µg/L) compared with those in the lower quartile of exposure (blood cadmium <0.25 µg/L) (µg/L, ß=-0.14, 95% CI -0.25 to -0.03), and there was a trend by quartiles of blood cadmium (P<0.0001). CONCLUSIONS: Our findings suggest that increased blood cadmium is associated with worse cognitive function in adults aged 60 years or older in the USA.

Chemical Constituents and Antioxidant, Anti-Inflammatory and Anti-Tumor Activities of Melilotus officinalis (Linn.) Pall.

Two new p-hydroxybenzoic acid glycosides, namely p-hydroxybenzoic acid-4-O-α-d-manopyranosyl-(1 → 3)-α-l-rhamnopyranoside (compound 1) and 4-O-α-l-rhamnopyran-osyl-(1 → 6)-α-d-manopyranosyl-(1 → 3)-α-l-rhamnopyranoside (compound 2), and seven known compounds, compound 3, 6, 7 (acid components), compound 8, 9 (flavonoids), compound 4 (a coumarin) and compound 5 (an alkaloid), were isolated from the 70% ethanol aqueous extract of the aerial parts of Melilotus officinalis (Linn.) Pall. The structures of all compounds were elucidated by use of extensive spectroscopic methods Infrared Spectroscopy (IR), High resolution electrospray ionization mass spectrometry (HR-ESI-MS), and ¹H and 13C-NMR). Sugar residues obtained after acid hydrolysis were identified by high-performance liquid chromatography (HPLC). The antioxidant activity of all the compounds was evaluated by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS⁺) and 1,1-diphenyl-2-picrylhydrazyl (DPPH). The anti-inflammatory effects of the compounds were also evaluated in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. All compounds were shown to inhibit LPS-induced nitric oxide (NO) and prostaglandin E 2 (PGE 2) production by suppressing the expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2), respectively, in LPS-stimulated RAW 264.7 cells. The inhibitory effect of all the compounds on MCF-7 cells was determined by Cell Counting Kit-8 (CCK-8) method. The results showed that compounds 1, 2, 7, 8, 9 exhibited better antioxidant activity compared to the other compounds. compounds 1-9 had different inhibitory effects on the release of NO, TNF-α and IL-6 in LPS-stimulated RAW264.7 cells by LPS, of which compound 7 was the most effective against inflammatory factors. compounds 1 and 2 have better antitumor activity compared to other compounds. Further research to elucidate the chemical composition and pharmacological effects of Melilotus officinalis (Linn.) Pall is of major importance towards the development and foundation of clinical application of the species.