Single cell transcriptomic analysis reveals tumor immune infiltration by NK cells gene signature in lung adenocarcinoma.

Background: Natural Killer (NK) cells are vital components of the innate immune system, crucial for combating infections and tumor growth, making them pivotal in cancer prognosis and immunotherapy. We sought to understand the diverse characteristics of NK cells within lung adenocarcinoma (LUAD) by conducting single-cell RNA sequencing analyses. Methods: Using the scRNA-seq dataset for multiple primary lung cancers (MPLCs), we examined two major NK cell groups, NK1 and NK2, comparing the expression profiles of 422 differentially expressed NK signature genes. We identified eight genes (SPON2, PLEKHG3, CAMK2N1, RAB27B, CTBP2, EFHD2, GOLM1, and PLOD1) that distinguish NK1 from NK2 cells. A prognostic signature, the NK gene signature (NKGS) score, was established through LASSO Cox regression. High NKGS scores were linked to poorer overall survival in TCGA-LUAD patients and consistently validated in other datasets (GSE31210 and GSE14814). Results: Functional analysis revealed an enrichment of genes related to the TGF-ß signaling pathway in the high NKGS score group. Moreover, a high NKGS score correlated with an immunosuppressive tumor microenvironment (TME) driven by immune evasion mechanisms. We also observed reduced T-cell receptor (TCR) repertoire diversity in the high-risk NKGS group, indicating a negative association between inflammation and risk score. Conclusion: This study introduced the innovative NKGS score, differentiating NK1 from NK2 cells. High NKGS scores were associated with the TGF-ß pathway and provided insights into LUAD prognosis and immune activities.

Plasma microRNA-320c as a Potential Biomarker for the Severity of Knee Osteoarthritis and Regulates cAMP Responsive Element Binding Protein 5 (CREB5) in Chondrocytes.

Objective: Osteoarthritis (OA) is a commonly known prevalent joint disease, with limited therapeutic methods. This study aimed to investigate the expression of plasma microRNA-320c (miR-320c) in patients with knee OA and to explore the clinical value and potential mechanism of miR-320c in knee OA. Methods: Forty knee OA patients and 20 healthy controls were enrolled. The levels of plasma miR-320c and plasma inflammatory cytokines were measured by real-time PCR or ELISA. Correlations of Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores and cytokine levels with the miR-320c expression level were evaluated by Pearson correlation analysis. Then, a receiver operating characteristic (ROC) curve was drawn to analyse the diagnostic value of miR-320c in OA. Finally, the interaction of miR-320c and cAMP responsive element binding protein 5 (CREB5) was determined using a luciferase reporter assay, and the effect of CREB5 on the cAMP pathway was assessed. Results: The expression level of plasma miR-320c was significantly higher in OA patients than in healthy controls (p < 0.05). The increased plasma miR-320c level was positively correlated with the WOMAC score (r = 0.796, p < 0.001) and the plasma interleukin (IL)-1ß (r = 0.814, p < 0.001) and IL-6 (r = 0.695, p < 0.001) levels in patients with OA. ROC curve analysis demonstrated the relatively high diagnostic accuracy of plasma miR-320c for OA. Furthermore, the luciferase reporter assay results showed that miR-320c regulates CREB5 expression by binding to the CREB5 3'-untranslated region. Moreover, suppression of CREB5 significantly reduced the expression levels of c-fos and c-jun. Conclusion: Our results indicate that plasma miR-320c may serve as a potential novel predictor of the severity of knee OA and that miR-320c may play an important role in the pathogenesis of OA through inhibiting the cAMP pathway by targeting CREB5.

Revisiting Dinuclear Ruthenium Water Oxidation Catalysts: Effect of Bridging Ligand Architecture on Catalytic Activity.

An attractive catalytic pathway for the conversion of water to oxygen would involve two metal oxide centers combining in a constructive sense to make O═O. This prospect makes the study of certain dinuclear transition metal complexes particularly attractive. In this work, we describe the design and synthesis of two symmetrical bis-tridentate polypyridine ligands 6 and 12 that bind two RuII centers at a separation of 3.6 Šin 7 and 5.7 Šin 13. In the presence of CeIV at pH = 1, these systems oxidize water with the system having the more proximal metals being more reactive. In the case of the more proximal metal centers, the bridging ligand is a 3,6-disubstituted pyridazine which, under the influence of CeIV, cleaves into two [Ru(bpc)(pic)2CH3CN]+ fragments (14) which then function as the actual catalyst (bpc = 2,2'-bipyridine-6-carboxylate, pic = 4-methylpyridine). The second dinuclear catalyst contains a central pyrimidine ring which is less sensitive to oxidative decay and hence less reactive. Caution is advised in the use of CeIV as a sacrificial electron acceptor due to unexpected oxidative decay of the catalyst.

MicroRNA-320c inhibits articular chondrocytes proliferation and induces apoptosis by targeting mitogen-activated protein kinase 1 (MAPK1).

AIM: To clarify the interaction of microRNA-320c (miR-320c) and mitogen-activated protein kinase 1 (MAPK1), and to investigate the effects of miR-320c on articular chondroctye proliferation and apoptosis. METHODS: Lentiviral expression vectors were constructed and dual luciferase assays containing MAPK1 3'-untranslated regions (3'-UTRs) were performed. Small hairpin RNA (shRNA) was utilized to modulate MAPK1 expression. The messenger RNA and protein expression levels were determined by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting respectively. Cell Counting Kit-8 and flow cytometry were conducted to detect the proliferation and apoptosis of Human Chondrocyte-articular (HC-a) cells. Besides that, the influences of miR-320c and MAPK1 on MAPK pathway activation were also evaluated. RESULTS: Our data identified MAPK1 as a direct target gene of miR-320c, and miR-320c can negatively regulate MAPK1 expression by directly binding to MAPK1 3'-UTR in HC-a cells. Further functional study displayed that miR-320c overexpression and MAPK1 shRNA significantly suppressed the proliferation of HC-a cells and promoted cell apoptosis. Meanwhile, MAPK1 shRNA could attenuate miR-320c inhibitor promotive effects on HC-a cell proliferation and reverse its inhibitory effect on cell apoptosis. MAPK1 overexpression could rescue the inhibitory effect of miR-320c on HC-a cell proliferation, and weaken the accelerating effect of miR-320c on cell apoptosis. However, neither miR-320c or MAPK1 shRNA regulate the expression of c-JUN, JNK and c-Fos. CONCLUSION: miR-320c inhibits articular chondrocyte proliferation and induces apoptosis by targeting MAPK1, suggesting that miR-320c perhaps participates in the pathogenesis of osteoarthritis and acts as a potential target for the therapeutic treatment of osteoarthritis.

Leukocytapheresis Therapy for Rheumatoid Arthritis: Results Compared with Control Trial.

CONTEXT: Rheumatoid arthritis (RA) is a chronic multisystem autoimmune disease, mainly characterized by synovitis and with symmetrical joint involvement. LCAP therapy for RA patients has been shown to be safe and efficacious in some developed countries for over a decade. OBJECTIVE: The study intended to evaluate the efficacy and safety of leukocytopheresis (LCAP) for treatment of rheumatoid arthritis (RA) and to study the influence of treatment on the levels of various serum cytokines. DESIGN: The study was a nonblinded, nonrandomized, controlled trial. SETTING: The study took place in the Department of Rheumatology and Immunology at Beijing Hospital at the National Center of Gerontology in Beijing, China. PARTICIPANTS: Participants were 51 patients with RA at the hospital with a 28-joint disease activity score (DAS28) exceeding the 3.20 needed to fulfill the classification criteria of the American College of Rheumatology (ACR). INTERVENTION: Participants were divided into 2 groups. One group (intervention group) received LCAP therapy (n = 20), while the control group (n = 31) received disease-modifying antirheumatic drugs (DMARDs). Patients receiving the LCAP therapy were treated using a Cellsorba column every 5 days for a total of 5 treatments. OUTCOME MEASURES: Clinical assessment of participants' symptoms included: (1) a tender-joint count, (2) a swollen-joint count, (3) erythrocyte sedimentation rates (ESR), (4) C-reactive protein levels (CRP), (5) a visual analog scale (VAS) for pain, (6) the DAS28 C-reactive protein (DAS28-CRP) scores, and the Health Assessment Questionnaire Disability Index (HAQ-DI). The study also evaluated participants' scores for the American College of Rheumatology (ACR) Core Data Set. Serum collected before and after therapy from both groups was analyzed for the levels of bradykinin, serotonin, heat shock protein 70, human CXC-chemokine ligand 16 (CXCL16), prostaglandin E2, and macrophage inflammation protein 1α. RESULTS: At week 4 for participants receiving the LCAP therapy, ACR20, ACR50, and ACR70 were observed in 55%, 30%, and 20% of patients, respectively, compared to 19.4%, 3.2%, and 0% for patients in the control group (P < .05). Also, at week 24 of LCAP therapy, ACR20, ACR50, and ACR70 were observed in 70%, 50%, and 30% of patients, respectively, which was significantly higher than the 25.8%, 12.9%, and 3.2% of patients in the control group (P < .05). The serum levels of CXCL16 and serotonin were significantly reduced in the LCAP group compared with control group. CONCLUSIONS: This study indicated that LCAP therapy can significantly decrease RA disease activity and is a safe and effective alternative therapy. LCAP therapy significantly reduced serum CXCL16 and serotonin levels, offering a putative mechanism by which it improves the articular symptoms of RA.

Iguratimod ameliorates bleomycin-induced alveolar inflammation and pulmonary fibrosis in mice by suppressing expression of matrix metalloproteinase-9.

AIM: To investigate the potential therapeutic efficacy of iguratimod (IGU) on bleomycin (BLM)-induced pulmonary fibrosis in mice. METHODS: A total of 75 C57BL/6 mice were randomly and evenly divided into control group, BLM (5 mg/kg) group, BLM + IGU (90 mg/kg) group, BLM + methylprednisolone (MP, 10 mg/kg) group and BLM + pirfenidone (PF, 100 mg/kg) group. The mice were sacrificed on day 7, 14 and 28. The lung tissue was examined by hematoxylin and eosin staining and Masson staining to evaluate the degree of alveolitis and fibrosis, and serum cytokines were measured. RESULTS: Histopathological results showed that IGU attenuated BLM-induced alveolar inflammation and decreased collagen deposition in lung tissue from day 7 till day 28. Both the pathological alveolitis and fibrosis scores in the drug-treated groups (IGU group, MP group and PF group) were decreased dramatically compared with the BLM group on day 7, 14 and 28 (P < 0.05). There were no statistical significances among these three groups. Cytokine profile showed that IGU decreased the level of tumor necrosis factor-α (TNF-α), interleukin (IL)-1, IL-6 and matrix metalloproteinase (MMP)-9 which were up-regulated by BLM on day 7, 14 and 28 (P < 0.05). Furthermore, there is a strong correlation between the severity of pulmonary fibrosis and serum MMP-9 levels. CONCLUSION: IGU can decrease BLM-induced pulmonary fibrosis, and the anti-fibrotic effect of IGU is mediated partly via inhibition of MMP-9, which suggests that IGU could potentially be an effective therapeutic strategy for pulmonary fibrosis.

Effect of colchicine in prevention of pericardial effusion and atrial fibrillation: a meta-analysis.

Randomized, controlled trials (RCTs) have assessed the effect of colchicine therapy in prevention of pericardial effusion (PE) and atrial fibrillation (AF). However, the effects are still inconclusive. PubMed, Cochrane Library, Google Scholar, and EMBASE database were searched. Primary outcome was the risk of PE and AF. Ten RCTs with 1981 patients and a mean follow-up of 12.6 months were included. Colchicine therapy was not associated with a significantly lower risk of post-operative PE (RR, 0.89; 95 % CI 0.70-1.13; p = 0.33, I (2) = 72.8 %) and AF (RR, 0.77; 95 % CI 0.52-1.13; p = 0.18, I (2) = 47.3 %). However, rates of pericarditis recurrence, symptoms persistence, and pericarditis-related hospitalization were significantly decreased with colchicine treatment. In addition, cardiac tamponade occurrence was similar between groups, and adverse events were significantly higher in the colchicine group. Colchicine may not significantly decrease the post-operative risk of PE and AF. However, only limited studies about patients undergoing cardiac surgery provide data about PE and AF.

Iron Complexes of Square Planar Tetradentate Polypyridyl-Type Ligands as Catalysts for Water Oxidation.

The tetradentate ligand, 2-(pyrid-2'-yl)-8-(1″,10″-phenanthrolin-2″-yl)-quinoline (ppq) embodies a quaterpyridine backbone but with the quinoline C8 providing an additional sp(2) center separating the two bipyridine-like subunits. Thus, the four pyridine rings of ppq present a neutral, square planar host that is well suited to first-row transition metals. When reacted with FeCl3, a µ-oxo-bridged dimer is formed having a water bound to an axial metal site. A similar metal-binding environment is presented by a bis-phenanthroline amine (dpa) which forms a 1:1 complex with FeCl3. Both structures are verified by X-ray analysis. While the Fe(III)(dpa) complex shows two reversible one-electron oxidation waves, the Fe(III)(ppq) complex shows a clear two-electron oxidation associated with the process H2O-Fe(III)Fe(III) → H2O-Fe(IV)Fe(IV) → O═Fe(V)Fe(III). Subsequent disproportionation to an Fe═O species is suggested. When the Fe(III)(ppq) complex is exposed to a large excess of the sacrificial electron-acceptor ceric ammonium nitrate at pH 1, copious amounts of oxygen are evolved immediately with a turnover frequency (TOF) = 7920 h(-1). Under the same conditions the mononuclear Fe(III)(dpa) complex also evolves oxygen with TOF = 842 h(-1).

Ruthenium catalysts for water oxidation involving tetradentate polypyridine-type ligands.

A series of Ru(II) complexes that behave as water oxidation catalysts were prepared involving a tetradentate equatorial ligand and two 4-substituted pyridines as the axial ligands. Two of these complexes were derived from 2,9-di-(pyrid-2'-yl)-1,10-phenanthroline (dpp) and examine the effect of incorporating electron-donating amino and bulky t-butyl groups on catalytic activity. A third complex replaced the two distal pyridines with N-methylimidazoles that are more electron-donating than the pyridines of dpp and potentially stabilize higher oxidation states of the metal. The tetradentate ligand 2-(pyrid-2'-yl)-6-(1'',10''-phenanthrol-2''-yl)pyridine (bpy-phen), possessing a bonding cavity similar to dpp, was also prepared. The Ru(II) complex of this ligand does not have two rotatable pyridines in the equatorial plane and thus shows different flexibility from the [Ru(dpp)] complexes. All the complexes showed activity towards water oxidation. Investigation of their catalytic behavior and electrochemical properties suggests that they may follow the same catalytic pathway as the prototype [Ru(dpp)pic2](2+) involving a seven-coordinated [Ru(IV)(O)] intermediate. The influence of coordination geometry on catalytic performance is analyzed and discussed.

Electrocatalytic H2 Evolution by Supramolecular Ru(II)-Rh(III)-Ru(II) Complexes: Importance of Ligands as Electron Reservoirs and Speciation upon Reduction.

The supramolecular water reduction photocatalysts [{(Ph2phen)2Ru(dpp)}2RhX2](PF6)5 (Ph2phen = 4,7-diphenyl-1,10-phenanthroline, dpp =2,3-bis(2-pyridyl)pyrazine X = Cl, Br) are efficient electrocatalysts for the reduction of CF3SO3H, CF3CO2H, and CH3CO2H to H2 in DMF or DMF/H2O mixtures. The onset of catalytic current occurs at -0.82 V versus Ag/AgCl for CF3SO3H, -0.90 V for CF3CO2H, and -1.1 V for CH3CO2H with overpotentials of 0.61, 0.45, and 0.10 V, respectively. In each case, catalysis is triggered by the first dpp ligand reduction implicating the dpp as an electron reservoir in catalysis. A new species with Epc ∼ -0.75 V was observed in the presence of stoichiometric amounts of strong acid, and its identity is proposed as the Rh(H)(III/II) redox couple. H2 was produced in 72-85% Faradaic yields and 95-116 turnovers after 2 h and 435 turnovers after 10 h of bulk electrolysis. The identities of Rh(I) species upon reduction have been studied. In contrast to the expected dissociation of halides in the Rh(I) state, the halide loss depends on solvent and water content. In dry CH3CN, in which Cl(-) is poorly solvated, a [Ru] complex dissociates and [(Ph2phen)2Ru(dpp)Rh(I)Cl2](+) and [(Ph2phen)2Ru(dpp)](2+) are formed. In contrast, for X = Br(-), the major product of reduction is the intact trimetallic Rh(I) complex [{(Ph2phen)2Ru(dpp)}2Rh(I)](5+). Chloride loss in CH3CN is facilitated by addition of 3 M H2O. In DMF, the reduced species is [{(Ph2phen)2Ru(dpp)}2Rh(I)](5+) regardless of X = Cl(-) or Br(-).

A new Ru(II)Rh(III) bimetallic with a single Rh-Cl bond as a supramolecular photocatalyst for proton reduction.

A new Ru(II)Rh(III) structural motif [(bpy)2Ru(dpp)RhCl(tpy)](4+) with one halide on the Rh(III) center demonstrates light-driven proton reduction ability, establishing that two halide ligands are not mandatory despite all prior systems containing a cis-RhCl2 catalytic site. This new design provides a novel approach to modulate Rh(III) redox behavior and catalytic activity with insight into catalytic intermediates.

Tuning the Photophysical Properties of Ru(II) Monometallic and Ru(II),Rh(III) Bimetallic Supramolecular Complexes by Selective Ligand Deuteration.

A series of three new complexes of the design [(TL)2Ru(BL)](2+), two new complexes of the design [(TL)2Ru(BL)Ru(TL)2](4+), and three new complexes of the design [(TL)2Ru(BL)RhCl2(TL)](3+) (TL = bpy or d8-bpy; BL = dpp or d10-dpp; TL = terminal ligand; BL = bridging ligand; bpy = 2,2'-bipyridine; dpp = 2,3-bis(2-pyridyl)pyrazine) were synthesized and the (1)H NMR spectroscopy, electrochemistry, electronic absorbance spectroscopy, and photophysical properties studied. Incorporation of deuterated ligands into the molecular architecture simplifies the (1)H NMR spectra, allowing for complete (1)H assignment of [(d8-bpy)2Ru(dpp)](PF6)2 and partial assignment of [(bpy)2Ru(d10-dpp)](PF6)2. The electrochemistry for the deuterated and nondeuterated species showed nearly identical redox properties. Electronic absorption spectroscopy of the deuterated and nondeuterated complexes are superimposable with the lowest energy transition being Ru(dπ) → BL(π*) charge transfer in nature (BL = dpp or d10-dpp). Ligand deuteration impacts the excited-state properties with an observed increase in the quantum yield of emission (Φ(em)) and excited-state lifetime (τ) of the Ru(dπ) → d10-dpp(π*) triplet metal-to-ligand charge transfer ((3)MLCT) excited state when dpp is deuterated, and a decrease in the rate constant for nonradiative decay (knr). Choice of ligand deuteration between bpy and dpp strongly impacts the observed photophysical properties with BL = d10-dpp complexes showing an enhanced Φ(em) and τ, providing further support that the lowest electronic excited state populated via UV or visible excitation is the photoactive Ru(dπ) → dpp(π*) CT excited state. The Ru(II),Rh(III) complex incorporating the deuterated BL shows increased hydrogen production compared to the variants incorporating the protiated BL, while demonstrating identical dynamic quenching behaviors in the presence of sacrificial electron donor.

New supramolecular structural motif coupling a ruthenium(II) polyazine light absorber to a rhodium(I) center.

Two new complexes, [(bpy)2Ru(dpp)Rh(I)(COD)](PF6)3 and [(Me2bpy)2Ru(dpp)Rh(I)(COD)](PF6)2(BF4) (bpy = 2,2'-bipyridine, Me2bpy = 4,4'-dimethyl-2,2'-bipyridine, dpp = 2,3-bis(2-pyridyl)pyrazine, and COD = 1,5-cyclooctadiene), representing a new Ru(II),Rh(I) structural motif, have been prepared and characterized by mass spectrometry, (1)H NMR spectroscopy, electrochemistry, electronic absorption spectroscopy, and emission spectroscopy. These two complexes represent a new type of supramolecular complex with a [(TL)2Ru(dpp)](2+) (TL = terminal ligand) light absorber (LA) coupled to a Rh(I) center and are models for Ru(II),Rh(I) intermediates in the photochemical reduction of water using dpp-bridged Ru(II),Rh(III) photocatalysts. Electrochemical study reveals overlapping reversible Ru(II/III) and irreversible Rh(I/II/III) oxidations and a quasi-reversible dpp(0/-) reduction, demonstrating that the lowest unoccupied molecular orbital (LUMO) is dpp(π*) based. The COD ligand is sterically bulky, displaying steric repulsions between hydrogen atoms on the alkene of COD and dpp about the square planar Rh(I) center. An interesting reactivity occurs in coordinating solvents such as CH3CN, where Rh(I) substitution leads to an equilibrium between the Ru(II),Rh(I) bimetallic and [(TL)2Ru(dpp)](2+) and [Rh(I)(COD)(solvent)2](+) monometallic species. The electronic absorption spectra of both complexes feature transitions at ca. 500 nm attributed to a Ru(dπ) → dpp(π*) metal-to-ligand charge transfer (MLCT) transition that is slightly red-shifted from the Ru synthon upon Rh(I) complexation. The methylation of TL on the Ru impacts the electrochemical and optical properties in a minor but predictable manner. The photophysical studies, by comparison with the model complex [{Ru(bpy)2}2(dpp)](PF6)4 and related Rh(III) complex [(bpy)2Ru(dpp)Rh(III)Cl2(phen)](PF6)3, reveal the expected absence of a Ru(dπ) → Rh(dσ*) (3)MMCT state (metal-to-metal charge transfer) in the title complexes, which is present in Rh(III) systems. The absence of this (3)MMCT state in Ru(II),Rh(I) complexes results in a longer lifetime and higher emission quantum yield for the Ru(dπ) → dpp(π*) (3)MLCT state than [(bpy)2Ru(dpp)Rh(III)Cl2(phen)](PF6)3. Both complexes display photocatalytic hydrogen production activity in the presence of water and a sacrificial electron donor, with the [(bpy)2Ru(dpp)Rh(I)(COD)](PF6)3 possessing a higher catalytic activity than the methyl analogue. Both display low activities, hypothesized to occur due to steric crowding about the Rh(I) site.