Syringic acid attenuates acute lung injury by modulating macrophage polarization in LPS-induced mice.

BACKGROUND: Acute lung injury (ALI) is a continuum of lung changes caused by multiple lung injuries, characterized by a syndrome of uncontrolled systemic inflammation that often leads to significant morbidity and death. Anti-inflammatory is one of its treatment methods, but there is no safe and available drug therapy. Syringic acid (SA) is a natural organic compound commonly found in a variety of plants, especially in certain woody plants and fruits. In modern pharmacological studies, SA has anti-inflammatory effects and therefore may be a potentially safe and available compound for the treatment of acute lung injury. PURPOSE: This study attempts to reveal the protective mechanism of SA against ALI by affecting the polarization of macrophages and the activation of NF-κB signaling pathway. Trying to find a safer and more effective drug therapy for clinical use. METHODS: We constructed the ALI model using C57BL/6 mice by intratracheal instillation of LPS (10 mg/kg). Histological analysis was performed with hematoxylin and eosin (H&E). The wet-dry ratio of the whole lung was measured to evaluate pulmonary edema. The effect of SA on macrophage M1-type was detected by flow cytometry. BCA protein quantification method was used to determine the total protein concentration in bronchoalveolar lavage fluid (BALF). The levels of Interleukin (IL)-6, IL-1ß, and tumor necrosis factor (TNF)-α in BALF were determined by the ELISA kits, and RT-qPCR was used to detect the expression levels of IL-6, IL-1ß and TNF-α mRNA of lung tissue. Western blot was used to detect the expression levels of iNOS and COX-2 and the phosphorylation of p65 and IκBα in the NF-κB pathway in lung tissue. In vitro experiments were conducted with RAW267.4 cell inflammation model induced by 100 ng/ml LPS and A549 cell inflammation model induced by 10 µg/ml LPS. The effects of SA on M1-type and M2-type macrophages of RAW267.4 macrophages induced by LPS were detected by flow cytometry. The toxicity of compound SA to A549 cells was detected by MTT method which to determine the safe dose of SA. The expressions of COX-2 and the phosphorylation of p65 and IκBα protein in NF-κB pathway were detected by Western blot. RESULTS: We found that the pre-treatment of SA significantly reduced the degree of lung injury, and the infiltration of neutrophils in the lung interstitium and alveolar space of the lung. The formation of transparent membrane in lung tissue and thickening of alveolar septum were significantly reduced compared with the model group, and the wet-dry ratio of the lung was also reduced. ELISA and RT-qPCR results showed that SA could significantly inhibit the production of IL-6, IL-1ß, TNF-α. At the same time, SA could significantly inhibit the expression of iNOS and COX-2 proteins, and could inhibit the phosphorylation of p65 and IκBα proteins. in a dose-dependent manner. In vitro experiments, we found that flow cytometry showed that SA could significantly inhibit the polarization of macrophages from M0 type macrophages to M1-type macrophages, while SA could promote the polarization of M1-type macrophages to M2-type macrophages. The results of MTT assay showed that SA had no obvious cytotoxicity to A549 cells when the concentration was not higher than 80 µM, while LPS could promote the proliferation of A549 cells. In the study of anti-inflammatory effect, SA can significantly inhibit the expression of COX-2 and the phosphorylation of p65 and IκBα proteins in LPS-induced A549 cells. CONCLUSION: SA has possessed a crucial anti-ALI role in LPS-induced mice. The mechanism was elucidated, suggesting that the inhibition of macrophage polarization to M1-type and the promotion of macrophage polarization to M2-type, as well as the inhibition of NF-κB pathway by SA may be the reasons for its anti-ALI. This finding provides important molecular evidence for the further application of SA in the clinical treatment of ALI.

Pharmacological mechanisms of sinomenine in anti-inflammatory immunity and osteoprotection in rheumatoid arthritis: A systematic review.

BACKGROUND: Sinomenine (SIN) is the main pharmacologically active component of Sinomenii Caulis and protects against rheumatoid arthritis (RA). In recent years, many studies have been conducted to elucidate the pharmacological mechanisms of SIN in the treatment of RA. However, the molecular mechanism of SIN in RA has not been fully elucidated. PURPOSE: To summarize the pharmacological effects and molecular mechanisms of SIN in RA and clarify the most valuable regulatory mechanisms of SIN to provide clues and a basis for basic research and clinical applications. METHODS: We systematically searched SciFinder, Web of Science, PubMed, China National Knowledge Internet (CNKI), the Wanfang Databases, and the Chinese Scientific Journal Database (VIP). We organized our work based on the PRISMA statement and selected studies for review based on predefined selection criteria. OUTCOME: After screening, we identified 201 relevant studies, including 88 clinical trials and 113 in vivo and in vitro studies on molecular mechanisms. Among these studies, we selected key results for reporting and analysis. CONCLUSIONS: We found that most of the known pharmacological mechanisms of SIN are indirect effects on certain signaling pathways or proteins. SIN was manifested to reduce the release of inflammatory cytokines such as Tumor necrosis factor-α (TNF-α), Interleukin-6 (IL-6), and IL-1ß, thereby reducing the inflammatory response, and apparently blocking the destruction of bone and cartilage. The regulatory effects on inflammation and bone destruction make SIN a promising drug to treat RA. More notably, we believe that the modulation of α7nAChR and the regulation of methylation levels at specific GCG sites in the mPGES-1 promoter by SIN, and its mechanism of directly targeting GBP5, certainly enriches the possibilities and the underlying rationale for SIN in the treatment of inflammatory immune-related diseases.

Sinomenine ameliorates collagen-induced arthritis in mice by targeting GBP5 and regulating the P2X7 receptor to suppress NLRP3-related signaling pathways.

Sinomenine (SIN) is an isoquinoline alkaloid isolated from Sinomenii Caulis, a traditional Chinese medicine used to treat rheumatoid arthritis (RA). Clinical trials have shown that SIN has comparable efficacy to methotrexate in treating patients with RA but with fewer adverse effects. In this study, we explored the anti-inflammatory effects and therapeutic targets of SIN in LPS-induced RAW264.7 cells and in collagen-induced arthritis (CIA) mice. LPS-induced RAW264.7 cells were pretreated with SIN (160, 320, 640 µM); and CIA mice were administered SIN (25, 50 and 100 mg·kg-1·d-1, i.p.) for 30 days. We first conducted a solvent-induced protein precipitation (SIP) assay in LPS-stimulated RAW264.7 cells and found positive evidence for the direct binding of SIN to guanylate-binding protein 5 (GBP5), which was supported by molecular simulation docking, proteomics, and binding affinity assays (KD = 3.486 µM). More importantly, SIN treatment markedly decreased the expression levels of proteins involved in the GBP5/P2X7R-NLRP3 pathways in both LPS-induced RAW264.7 cells and the paw tissue of CIA mice. Moreover, the levels of IL-1ß, IL-18, IL-6, and TNF-α in both the supernatant of inflammatory cells and the serum of CIA mice were significantly reduced. This study illustrates a novel anti-inflammatory mechanism of SIN; SIN suppresses the activity of NLRP3-related pathways by competitively binding GBP5 and downregulating P2X7R protein expression, which ultimately contributes to the reduction of IL-1ß and IL-18 production. The binding specificity of SIN to GBP5 and its inhibitory effect on GBP5 activity suggest that SIN has great potential as a specific GBP5 antagonist.

High-Throughput Colorimetric Analysis of Nanoparticle-Protein Interactions Based on the Enzyme-Mimic Properties of Nanoparticles.

While an in-depth understanding of the biological behavior of engineered nanoparticles (NPs) is of great importance for their various applications, it remains challenging to quantitatively characterize NP-protein interactions in a simple and high-throughput manner. In the present work, we propose a new, colorimetric approach capable of quantitatively analyzing the adsorption of proteins onto the surface of NPs by their distinct peroxidase-mimic properties. Taking cationic AuNPs as an example, we demonstrate that this colorimetric method is capable of evaluating NP-protein interactions in a simple and high-throughput manner in multiwell plates. Important binding parameters (e.g., the binding affinity) of three different serum proteins (bovine serum albumin, transferrin, and lysozyme) as well as human serum to AuNPs with three different sizes (average diameters of 5, 10, and 15 nm) have been obtained. Based on a quantitative analysis of NP-protein interactions, we observe that the binding affinity and the inhibition efficiency of the nanozyme activity of AuNPs are strongly affected by the characteristics of proteins as well as the sizes of NPs. These results illustrate the great potential of the present colorimetric method as a simple, low-cost, and high-throughput platform for quantitatively investigating NP-protein interactions.

Risk Interval of Complete Blood Counts May be Closely Associated with Bone Mineral Density in the Elderly Chinese Population.

BACKGROUND: Accumulating research suggests that hematopoiesis and bone metabolism are interconnected. Several studies have investigated the partial indexes of peripheral blood counts related to bone mineral density (BMD). The aim of this study was to investigate the associations between all of the parameters, especially the risk interval of complete blood counts (CBC) and BMD in a sample of elderly subjects aged >70 years. METHODS: Three hundred and eighty-six subjects aged > 70 years in our hospital were enrolled in a cross-sectional study and underwent BMD measurement along with a CBC test. Patients were divided into two groups: "at least osteopenia" (T-score < -1) and a normal group (T-score ≥ -1). The clinicopathological characteristics, CBC parameters, and BMD were analyzed between the two groups. We performed a supervised discretization (using a conditional inference tree algorithm) to find the risk interval for the continuous variables, especially for CBC parameters, and bootstrap multivariable logistic regression to estimate the odds of CBC parameters associated with BMD. RESULTS: A total of 248 subjects were included in the study and divided into the normal (n = 43) and "at least osteopenia" groups (n = 205). Subjects in the "at least osteopenia" group had varying degrees of decreases in white blood cell (WBC) count, red blood cell (RBC) count, hemoglobin (Hb), mean corpuscular hemoglobin concentration (MCHC), hematocrit (HCT), platelet volume distribution width (PDW) mean platelet volume (MPV), eosinophils, and lymphocytes, and had increases in platelets (PLTs). MCHC, WBC, RBC, PDW, MPV, Hb, and lymphocytes were successfully divided into two (low and high) intervals. Bootstrap logistic regression showed that low levels of body mass index (BMI) [(11.88, 23.53); OR: 4.07; p < 0.0001], lymphocytes [(0.54, 2.3); OR: 3.95; p < 0.0001] and PDW [(8.5, 12.7); OR: 2.44; p < 0.0001] along with being female and older age [(72, 97); OR: 2.16; p < 0.0001] were significantly associated with BMD as risk factors. CONCLUSIONS: The elderly with BMD loss tended to show an abnormal sign in the CBC test. Low levels of lymphocytes and PDW may contribute to the evaluation of osteoporosis risk in the elderly. Bone remodeling and hematopoiesis may have stronger associations and interactions than has been previously recognized.

Jingui Shenqi Pills Regulate Bone-Fat Balance in Murine Ovariectomy-Induced Osteoporosis with Kidney Yang Deficiency.

Jingui Shenqi Pills (JGSQP) have been a staple of traditional Chinese medicine for thousands of years, used primarily as a treatment for kidney yang deficiency (KYD). In vitro analyses of JGSQP revealed strong induction of osteogenic differentiation and inhibition of adipogenic differentiation in bone-marrow-derived mesenchymal stem/stromal cells. However, the mechanisms by which JGSQP regulate the bone-fat balance in murine ovariectomy-induced osteoporosis with KYD have not been reported. Materials and Methods. Two-month-old female C57BL/6 mice were divided randomly into three groups: those receiving a sham operation (Sham); those undergoing bilateral ovariectomy and selection of KYD syndrome (Model); and those subjected to both bilateral ovariectomy and KYD syndrome selection for 8 weeks, followed by JGSQP treatment for 4 weeks (JGSQP). In the Sham and Model groups, mice were given the same dose of distilled water orally for 4 weeks. Animals from all three groups were euthanised at the 12th week. Vertebral microarchitecture and histomorphology were examined by micro-CT and H&E staining, respectively. In addition, we examined the mRNA expression of Akt, Wnt10b, Osterix (Osx), Fndc5, PPARγ, and Fabp4, as well as the protein of AKT, phosphorylation-AKT (p-AKT), BMP2, COL1A1, and FNDC5. Results. JGSQP treatment improved bone microarchitecture and mitigated histomorphological damage relative to the Model group. The osteoblast number (Ob.N/BS) and area (Ob.S/BS) were increased, whereas adipocyte number (adipocyte/tissue area) and area (adipocyte area/tissue area) were decreased in the JGSQP group. JGSQP treatment reduced the mRNA expression of Akt and adipogenesis-related genes (Fndc5, PPARγ, and Fabp4) while promoting osteogenesis-related genes (Wnt10b and Osx) mRNA expression. Additionally, the expression of p-AKT, BMP2, and COL1A1 proteins was increased and FNDC5 protein expression was decreased after JGSQP treatment. Conclusions. JGSQP treatment reversed murine ovariectomy-induced osteoporosis with KYD by controlling bone-fat balance via AKT pathway.

In Situ Investigation on the Protein Corona Formation of Quantum Dots by Using Fluorescence Resonance Energy Transfer.

A fundamental understanding of nanoparticle-protein corona and its interactions with biological systems is essential for future application of engineered nanomaterials. In this work, fluorescence resonance energy transfer (FRET) is employed for studying the protein adsorption behavior of nanoparticles. The adsorption of human serum albumin (HSA) onto the surface of InP@ZnS quantum dots (QDs) with different chirality (d- and l-penicillamine) shows strong discernible differences in the binding behaviors including affinity and adsorption orientation that are obtained upon quantitative analysis of FRET data. Circular dichroism spectroscopy further confirms the differences in the conformational changes of HSA upon interaction with d- and l-chiral QD surfaces. Consequently, the formed protein corona on chiral surfaces may affect their following biological interactions, such as possible protein exchange with serum proteins plasma as well as cellular interactions. These results vividly illustrate the potential of the FRET method as a simple yet versatile platform for quantitatively investigating biological interactions of nanoparticles.

Novel perylene probe-encapsulated metal-organic framework nanocomposites for ratiometric fluorescence detection of ATP.

Adenosine triphosphate (ATP) plays an important role in various biological processes and the ATP level is closely associated with many diseases. Herein, a novel ratiometric fluorescence assay for ATP was developed based on the excimer-monomer transfer of a perylene probe. By encapsulating a perylene probe, N,N'-bis(6-caproic acid)-3,4:9,10-perylenediimide (PDI), into zeolitic imidazolate framework-8 (ZIF-8) nanocrystals, fluorescent nanocomposites (PDI@ZIF-8) with significant excimer emission of the perylene probe were prepared for the first time. The presence of ATP will trigger the decomposition of PDI@ZIF-8 due to much stronger coordination between ATP and Zn2+ than that of 2-methylimidazole and Zn2+. As a result, the encapsulated PDI probes were released, leading to significantly increased monomer emission accompanying the decrease in the excimer emission. The excimer-monomer transition signal was utilized for ratiometric ATP sensing and its potential application for detecting ATP in cell lysates was also successfully demonstrated.

Mobile phone application for self-assessment of acute stroke patients: A tool for extended care and follow-up.

We aimed to assess the accuracy of self-assessment for acute stroke patients via mobile phone application-based scales and determine the value and prospect of clinical use.A cross-sectional study was designed and acute stroke patients were enrolled. We pushed the modified Rankin scale (mRS) and activities of daily living (ADL) scale to patients via mobile phone application for self-assessment on the day before they were out of hospital. We compared the results from nurse assessment and self-assessment.Around 50 patients with the average age 51.72 ±â€Š12.40 completed the self-assessment. A total of 27 patients self-assessed the scales, while caregivers of other 23 patients completed the assessment. In comparison with patient assessment and nurse assessment, significant difference was found in ADL score (P = .004), but was not found in mRS score (P > .05). When comparing caregiver assessment with nurse assessment, no significant difference could be found either in ADL score (P > .05) or in mRS score (P > .05). The kappa value for self-assessment and nurse agreement of ADL was 0.720 (P = .000), with sensitivity 96.8% and specificity 82.0%. The kappa value for self-assessment and nurse agreement of mRS was 0.718 (P = .000), with sensitivity 97.6% and specificity 92.4%.In summary, mobile phone application-based scales are generally accurate, economical and convenient for self-assessment of acute stroke patients with acceptable reliability in our small scale study. Caregivers can serve as the proper assessor when patients are out of hospital. Therefore, it is promising but still need to be further confirmed how practical to use this application in extended care and follow-up.

Tuning of the perylene probe excimer emission with silver nanoparticles.

Silver nanoparticles (Ag NPs) enhanced perylene probe excimer emission is reported for the first time. It was observed that strong interactions between the perylene probe and the Ag NPs induced co-aggregation. As a result, a new in situ generated plasmonic absportion band of the Ag NPs at longer wavelength emerged. The monomer emission of the perylene probe was efficiently quenched, and dramatically enhanced probe excimer emission was observed. A remarkable emission enhancement of over 1000 fold was obtained compared to anionic polymers and other nanoparticles. The excimer emission intensity could be finely modulated by the size of the Ag NPs and the functionalities of the perylene probe. The observed Ag NPs enhanced perylene probe excimer emission shows good potential for the development of novel sensing techniques for various bioanalytical applications.

Peroxidase activity of the coronene bisimide supramolecular architecture and its applications in colorimetric sensing of H2O2 and glucose.

A new water soluble coronene bisimide derivative (CTDI) was designed and synthesized. CTDI self-assembled in an aqueous solution and formed supramolecular nanofibers through π-π stacking and hydrophobic interactions. The nanofibers exhibit distinct peroxidase-like catalytic activity, and can catalyze the redox reaction of 3,3,5,5-tetramethylbenzidine (TMB) in the presence of hydrogen peroxide. Clear assay solution color changes were observed. The peroxidase-like catalytic property was utilized for the sensitive detection of H2O2 and glucose. The assay shows excellent sensitivity, and 1 µM of glucose could be easily detected. Glucose detection in dilute human blood samples was also demonstrated, and the results were verified using a commercial glucose meter. Our method is simple, convenient, sensitive and selective, and could facilitate the sensing of glucose in relation to biological and biomedical research applications.

One-pot Synthesis of Quencher Labeled Hairpin DNA-CdTe QDs Conjugate for Target DNA and Deoxyribonuclease I Detection.

A black-hole quencher (BHQ-2) labeled DNA (Q-DNA) with a phosphorothioate backbone was covalently conjugated to the CdTe QDs during the QDs synthesis procedure. The hairpin structure of Q-DNA shortened the distance of the CdTe QDs and the BHQ-2 group, which resulted in fluorescence quenching of the QDs. The addition of target DNA or deoxyribonuclease I (DNase I) could move the BHQ-2 group away from the QDs. As a result, the fluorescence of the CdTe QDs recovered. This work provides a new way for target DNA and DNase I detection.

Fluorescence turn-on detection of mercury ions based on the controlled adsorption of a perylene probe onto the gold nanoparticles.

A novel fluorescence turn-on strategy based on Au nanoparticles and a perylene probe for the sensing of Hg(2+) ions has been developed. It was observed that a perylene probe could be adsorbed onto the surface of Au NPs through strong electrostatic and hydrophobic interactions. Its fluorescence was efficiently quenched by the Au nanoparticles. However, in the presence of Hg(2+) and NaBH4, Hg(2+) was reduced and an Au/Hg amalgam was formed on the surface of the Au nanoparticles. The perylene probe could hardly be adsorbed and quenched by the Au/Hg amalgam. A turn on fluorescence signal was therefore detected. The assay is quite sensitive, and 5 nM Hg(2+) could be easily detected. It is also very selective, a number of metal ions were tested and no noticeable interference was observed. The assay was also successfully applied for the determination of Hg(2+) in lake water samples. A simple, fast, inexpensive, highly sensitive and selective Hg(2+) sensing strategy is therefore established.

Polycation-induced benzoperylene probe excimer formation and the ratiometric detection of heparin and heparinase.

A benzoperylene probe excimer emission in an aqueous buffer solution is observed for the first time, and a novel ratiometric fluorescence method based on the probe excimer emission for the sensitive detection of heparin and heparinase is demonstrated. A negatively charged benzoperylene derivative, 6-(benzo[ghi]perylene-1,2-dicarboxylic imide-yl)hexanoic acid (BPDI), was employed. A polycation, poly(diallyldimethylammonium) chloride (poly-DDA), could induce aggregation of BPDI through noncovalent interactions. A decrease of BPDI monomer emission and a simultaneous increase of BPDI excimer emission were observed. Upon the addition of heparin, the strong binding between heparin and poly-DDA caused release of BPDI monomer molecules, and an excimer-monomer emission signal transition was detected. However, after the enzymatic hydrolysis of heparin by heparinase, heparin was hydrolyzed into small fragments, which weakened the competitive binding of heparin to poly-DDA. Poly-DDA induced aggregation of BPDI, and a monomer-excimer emission signal transition was detected. Our assay is simple, rapid, inexpensive, sensitive and selective, which could facilitate the heparin and heparinase related biochemical and biomedical research.

Label-free fluorescence turn-on detection of microRNA based on duplex-specific nuclease and a perylene probe.

A novel fluorescence turn-on microRNA (miRNA) detection method based on duplex-specific nuclease (DSN) and a perylene probe is presented in this study. A positively charged perylene derivative (compound 1) was used as the fluorescent probe. Compound 1 exhibits strong monomer fluorescence in an aqueous buffer solution. It is well known that single-stranded DNA is a polyanion in nature. Thus, it can induce the aggregation of compound 1 through strong electrostatic, hydrophobic and π-π stacking interactions. As a result, the fluorescence of compound 1 was efficiently quenched. When the target miRNA was added, the formation of DNA-RNA hybridized duplex initiated the cleavage of the DNA strand by DSN cycle reaction, which resulted in disaggregation of compound 1. A fluorescence turn-on signal was detected, and a novel miRNA sensing method was therefore established. The presented method is label-free, simple, cost effective, sensitive and selective.

Fluorescence turn-on detection of glucose via the Ag nanoparticle mediated release of a perylene probe.

A novel fluorescence turn-on strategy for glucose sensing is demonstrated. The fluorescence of a perylene probe could be quenched by the silver nanoparticles (Ag NPs). The Ag NPs could be etched by H2O2 generated from the enzymatic oxidation of glucose. And efficient probe fluorescence recovery was detected.