Inhibition of Hmbox1 Promotes Cardiomyocyte Survival and Glucose Metabolism Through Gck Activation in Ischemia/Reperfusion Injury.

BACKGROUND: Exercise-induced physiological cardiac growth regulators may protect the heart from ischemia/reperfusion (I/R) injury. Homeobox-containing 1 (Hmbox1), a homeobox family member, has been identified as a putative transcriptional repressor and is downregulated in the exercised heart. However, its roles in exercise-induced physiological cardiac growth and its potential protective effects against cardiac I/R injury remain largely unexplored. METHODS: We studied the function of Hmbox1 in exercise-induced physiological cardiac growth in mice after 4 weeks of swimming exercise. Hmbox1 expression was then evaluated in human heart samples from deceased patients with myocardial infarction and in the animal cardiac I/R injury model. Its role in cardiac I/R injury was examined in mice with adeno-associated virus 9 (AAV9) vector-mediated Hmbox1 knockdown and in those with cardiac myocyte-specific Hmbox1 ablation. We performed RNA sequencing, promoter prediction, and binding assays and identified glucokinase (Gck) as a downstream effector of Hmbox1. The effects of Hmbox1 together with Gck were examined in cardiomyocytes to evaluate their cell size, proliferation, apoptosis, mitochondrial respiration, and glycolysis. The function of upstream regulator of Hmbox1, ETS1, was investigated through ETS1 overexpression in cardiac I/R mice in vivo. RESULTS: We demonstrated that Hmbox1 downregulation was required for exercise-induced physiological cardiac growth. Inhibition of Hmbox1 increased cardiomyocyte size in isolated neonatal rat cardiomyocytes and human embryonic stem cell-derived cardiomyocytes but did not affect cardiomyocyte proliferation. Under pathological conditions, Hmbox1 was upregulated in both human and animal postinfarct cardiac tissues. Furthermore, both cardiac myocyte-specific Hmbox1 knockout and AAV9-mediated Hmbox1 knockdown protected against cardiac I/R injury and heart failure. Therapeutic effects were observed when sh-Hmbox1 AAV9 was administered after I/R injury. Inhibition of Hmbox1 activated the Akt/mTOR/P70S6K pathway and transcriptionally upregulated Gck, leading to reduced apoptosis and improved mitochondrial respiration and glycolysis in cardiomyocytes. ETS1 functioned as an upstream negative regulator of Hmbox1 transcription, and its overexpression was protective against cardiac I/R injury. CONCLUSIONS: Our studies unravel a new role for the transcriptional repressor Hmbox1 in exercise-induced physiological cardiac growth. They also highlight the therapeutic potential of targeting Hmbox1 to improve myocardial survival and glucose metabolism after I/R injury.

Insight into a Target-Induced Photocurrent-Polarity-Switching Photoelectrochemical Immunoassay for Ultrasensitive Detection of Escherichia coli O157:H7.

Sensitive, portable methods of detection for foodborne pathogens hold great significance for the early warning and prevention of foodborne diseases and environmental pollution. Restricted by a complicated matrix and limited signaling strategies, developing a ready-to-use sensing platform with ultrahigh sensitivity remains challenging. In this work, near-infrared (NIR) light-responsive AgBiS2 nanoflowers (NFs) and Cu2O nanocubes (NCs) were introduced to construct a novel target-induced photocurrent-polarity-switchable system and verified for the development of an all-in-one, ready-to-use photoelectrochemical (PEC) immunosensor. NIR-responsive n-type AgBiS2 NFs and p-type Cu2O NCs producing anodic and cathodic photocurrents were conjugated with monoclonal (MAb1) and polyclonal antibodies (PAb2), respectively. Using a sandwich-type immunocomplex bridged by Escherichia coli O157:H7, an efficient photocurrent-polarity-switching PEC system was formed on a paper-based working electrode (PWE). Owing to the spatial separation of the photogenerated carriers and the elimination of false-positive/negative signals by the polarity-switchable photocurrent, the proposed NIR PEC immunoassay for E. coli O157:H7 exhibits a considerably low detection limit of 8 colony-forming units/milliliter (CFU/mL) with a linear range from 25 to 5 × 107 CFU/mL. The platform includes a PWE with an automatic cleaning function and a portable PEC analyzer with smartphone-compatible Bluetooth capability, thus achieving point-of-care testing of E. coli O157:H7. The sensor was applied to the analysis of pork samples artificially contaminated with E. coli O157:H7, and the detection results were in good agreement with the plate counting method, a gold standard in the field. This work aimed to investigate the photoelectric activity of the NIR-responsive p/n-type composites and to provide a new signal-reversal route for the construction of an all-in-one ready-to-use PEC immunosensor for the detection of low-concentration biomolecules.

Conjugation Length-Dependent Raman Scattering Intensity of Conjugated Polymers.

Polydiacetylenes, as a class of conjugated polymers with alternating conjugated C═C and C≡C bonds, have emerged as a promising probe material for biomedical Raman imaging, given their ultrastrong Raman scattering intensity. However, the relationship between the structure, especially the molecular length of polydiacetylenes, and their Raman scattering intensity remains unclear. In this work, a series of water-soluble polydiacetylenes, namely poly(deca-4,6-diynedioic acid) (PDDA) with different molecular weights (MWs), is prepared through controlled polymerization and degradation. The ultraviolet-visible (UV-vis) absorption spectroscopic and Raman spectroscopic studies on these polymers reveal that the Raman scattering intensity of PDDA increases nonlinearly with the MW. The MW-Raman scattering intensity relationship in the polymerization process is completely different from that in the degradation process. In contrast, the Raman scattering intensity increases more linearly with the maximal absorbance of the polymer, and the relationship between the Raman scattering intensity and the maximal absorbance of PDDA in the polymerization process is consistent with that in the degradation process. The Raman scattering intensity of PDDA hence exhibits a better dependence on the effective conjugation length of the polymer, which should guide the future design of conjugated polymers for Raman imaging applications.

Continuously Multiplexed Ultrastrong Raman Probes by Precise Isotopic Polymer Backbone Doping for Multidimensional Information Storage and Encryption.

Raman-based super multiplexing has attracted great interest in imaging, biological analysis, identity security, and information storage. It still remains a great challenge to synthesize a large number of different Raman-active molecules to fulfill the Raman color palette. Here, we report a facile and systematic strategy to construct continuously multiplexed ultrastrong Raman probes. By precisely incorporating different ratios of 13C isotope into the backbone of poly(deca-4,6-diynedioic acid) (PDDA), we can obtain a library of PDDAs with tunable double-bond Raman frequencies and adjustable intensity ratios of two triple-bond (13C≡13C and 12C≡12C) Raman peaks, while retaining the ultrastrong Raman signals and physicochemical properties of the polymer. We also demonstrate the successful application of 13C-doped PDDAs as security inks to generate a novel 3D matrix barcode system for information encryption and high-density data storage. The isotopically doped PDDA series herein pave a new way to advance Raman-based super multiplexing for diverse applications.

Nanomaterials Based on Functional Polymers for Sensitizing Cancer Radiotherapy.

Despite being the mainstay treatment for many types of cancer in clinic, radiotherapy is undertaking great challenges in overcoming a series of limitations. Radiosensitizers are promising agents capable of depositing irradiation energy and generating free radicals to enhance the radiosensitivity of tumor cells. Combining radiosensitizers with functional polymer-based nanomaterials holds great potential to improve biodistribution, circulation time, and stability in vivo. The derived polymeric nano-radiosensitizers can significantly improve the efficiency of tumor targeting and radiotherapy, and reduce the side effect to healthy tissues. In this review, an overview of functional polymer-based nanomaterials for radiosensitization in recent years is provided. Particular emphases are given to the action mechanisms, drug loading methods, targeting efficiencies, the impact on therapeutic effects, and biocompatibility of various radiosensitizing polymers, which are classified as polymeric micelles, dendrimers, polymeric nanospheres, nanoscale coordination polymers, polymersomes, and nanogels. The challenges and outlooks of polymeric nano-radiosensitizers are also discussed.

Complete Degradation of a Conjugated Polymer into Green Upcycling Products by Sunlight in Air.

The rapid development of digital society and artificial intelligence has triggered explosive demands for specialty plastics, especially conjugated polymers that are instrumental for flexible electronics and smart devices. The recycling and degradation of postconsumer conjugated polymers have become more important than ever to reduce the pressure to the environment. Here we report the discovery of an environmentally self-degradable conjugated polymer poly(deca-4,6-diynedioic acid), or PDDA. PDDA is stable in the dark or without oxygen when used as a functional material. However, when exposed to sunlight and air after the service life, PDDA disintegrates rapidly and fully decomposes through photooxidation in a week, yielding biocompatible, value-added succinic acid as a major degradation product. The complete degradation of PDDA into green upcycling products by sunlight in air, without leaving any microplastics, not only renders a pioneering paradigm of environmentally self-degradable conjugated polymers but also inspires developing effective strategies to completely degrade postconsumer conjugated polymers in a natural environment.

Fusobacterium nucleatum Increases Proliferation of Colorectal Cancer Cells and Tumor Development in Mice by Activating Toll-Like Receptor 4 Signaling to Nuclear Factor-κB, and Up-regulating Expression of MicroRNA-21.

BACKGROUND & AIMS: Nearly 20% of the global cancer burden can be linked to infectious agents. Fusobacterium nucleatum promotes tumor formation by epithelial cells via unclear mechanisms. We aimed to identify microRNAs (miRNAs) induced by F nucleatum and evaluate their ability to promote colorectal carcinogenesis in mice. METHODS: Colorectal cancer (CRC) cell lines were incubated with F nucleatum or control reagents and analyzed in proliferation and would healing assays. HCT116, HT29, LoVo, and SW480 CRC cell lines were incubated with F nucleatum or phosphate-buffered saline (PBS [control]) and analyzed for miRNA expression patterns and in chromatin immunoprecipitation assays. Cells were incubated with miRNAs mimics, control sequences, or small interfering RNAs; expression of reporter constructs was measured in luciferase assays. CRC cells were incubated with F nucleatum or PBS and injected into BALB/C nude mice; growth of xenograft tumors was measured. C57BL adenomatous polyposis colimin/+, C57BL miR21a-/-, and C57BL mice with full-length miR21a (controls) were given F nucleatum by gavage; some mice were given azoxymethane and dextran sodium sulfate to induce colitis and colon tumors. Intestinal tissues were collected and tumors were counted. Serum samples from mice were analyzed for cytokine levels by enzyme-linked immunosorbent assay. We performed in situ hybridization analyses to detect enrichment of F nucleatum in CRC cells. Fusobacterium nucleatum DNA in 90 tumor and matched nontumor tissues from patients in China were explored for the expression correlation analysis; levels in 125 tumor tissues from patients in Japan were compared with their survival times. RESULTS: Fusobacterium nucleatum increased proliferation and invasive activities of CRC cell lines compared with control cells. CRC cell lines infected with F nucleatum formed larger tumors, more rapidly, in nude mice than uninfected cells. Adenomatous polyposis colimin/+ mice gavaged with F nucleatum developed significantly more colorectal tumors than mice given PBS and had shorter survival times. We found several inflammatory factors to be significantly increased in serum from mice given F nucleatum (interleukin 17F, interleukin 21, and interleukin 22, and MIP3A). We found 50 miRNAs to be significantly up-regulated and 52 miRNAs to be significantly down-regulated in CRCs incubated with F nucleatum vs PBS; levels of miR21 increased by the greatest amount (>4-fold). Inhibitors of miR21 prevented F nucleatum from inducing cell proliferation and invasion in culture. miR21a-/- mice had a later appearance of fecal blood and diarrhea after administration of azoxymethane and dextran sodium sulfate, and had longer survival times compared with control mice. The colorectum of miR21a-/- mice had fewer tumors, of smaller size, and the miR21a-/- mice survived longer than control mice. We found RASA1, which encodes an RAS GTPase, to be one of the target genes consistently down-regulated in cells that overexpressed miR21 and up-regulated in cells exposed to miR21 inhibitors. Infection of cells with F nucleatum increased expression of miR21 by activating Toll-like receptor 4 signaling to MYD88, leading to activation of the nuclear factor-κB. Levels of F nucleatum DNA and miR21 were increased in tumor tissues (and even more so in advanced tumor tissues) compared with non-tumor colon tissues from patients. Patients whose tumors had high amounts of F nucleatum DNA and miR21 had shorter survival times than patients whose tumors had lower amounts. CONCLUSIONS: We found infection of CRC cells with F nucleatum to increase their proliferation, invasive activity, and ability to form xenograft tumors in mice. Fusobacterium nucleatum activates Toll-like receptor 4 signaling to MYD88, leading to activation of the nuclear factor-κB and increased expression of miR21; this miRNA reduces levels of the RAS GTPase RASA1. Patients with both high amount of tissue F nucleatum DNA and miR21 demonstrated a higher risk for poor outcomes.

Micro Integral Membrane Protein (MIMP), a Newly Discovered Anti-Inflammatory Protein of Lactobacillus Plantarum, Enhances the Gut Barrier and Modulates Microbiota and Inflammatory Cytokines.

BACKGROUND/AIMS: Recent studies have demonstrated that the manipulation of the gut microbiome represents a promising treatment for inflammatory bowel disease (IBD). We previously identified micro integral membrane protein (MIMP) as the smallest domain of surface layer protein from Lactobacillus Plantarum. However, the therapeutic relevance of MIMP in IBD remains unknown. METHODS: We initially employed a dextran sodium sulphate (DSS)-induced colitis model and evaluated the effect of MIMP on the inflammation response, intestinal barrier and gut microbiota using histological examination, Fluorescein isothiocyanate-Dextran detection and pyrosequencing analysis respectively. We then established peripheral blood mononuclear cells (PBMCs) and an epithelial CaCO-2 co-culture model to investigate the regulatory role of MIMP in inflammatory cytokines. The level changes of inflammatory cytokines were detected using Enzyme-linked immunosorbent and real-time polymerase chain reaction assay. The involved regulatory mechanisms were investigated mainly using dual luciferase reporter and chromatin immunoprecipitation assay. RESULTS: In the DSS-induced colitis model, we observed that MIMP intervention effectively improved the body weight loss, increased the colon length and decreased disease activity index. Consistently, the inflammation scores in the MIMP treatment group were significantly lower than those in the DSS treatment group. Furthermore, MIMP intervention was found to successfully neutralize DSS treatment by decreasing the expression of pro-inflammatory cytokines (IFN-γ, IL-17 and IL-23) and increasing the expression of anti-inflammatory cytokines (IL-4 and IL-10). Notably, the permeability assay demonstrated that the MIMP treatment group was remarkably lower than that in the DSS treatment group. We also showed that MIMP improved gut microbiota dysbiosis caused by DSS-induced inflammation. Additionally, in PBMCs and the CaCO-2 co-culture model, MIMP showed an obvious suppressive effect on lipopolysaccharide-induced inflammation in a time- and dose-dependent manner. Furthermore, we revealed that MIMP could modulate inflammatory cytokine expression through the toll-like receptor 4 pathway and histone acetylation. CONCLUSIONS: Our results suggested that MIMP showed a significant anti-inflammatory effect through regulating the gut barrier, microbiota and inflammatory cytokines. MIMP may have translational relevance as clinically relevant therapy for IBD patients.

Coordination environment dependence of anticancer activity in cyclometalated bismuth(III) complexes with C,O-chelating ligands.

In this paper, a series of cyclometalated bismuth(III) complexes bearing C,O-bidentate ligands were synthesized and characterized by techniques such as UV-vis, NMR, HRMS, and single crystal X-ray diffraction. Meanwhile, their cytotoxicities against various human cell lines, including colon cancer cells (HCT-116), breast cancer cells (MDA-MB-231), lung cancer cells (A549), gastric cancer cells (SGC-7901), and normal embryonic kidney cells (HEK-293) were assessed in vitro. Compared with the clinical cisplatin, most of the synthesized complexes possessed significantly higher degrees of anticancer activity and selectivity, giving a selectivity index of up to 71.3. The structure-activity relationship study revealed that the anticancer performance of these bismuth(III) species depends on the factors of coordination environment surrounding the metal center, such as coordination number, coordination bonding strength, lone 6s2 electron pair stereoactivity. The Annexin V-FITC/PI double staining assay results suggested that the coordination environment-dependent cytotoxicity is ascribable to apoptosis. Western blot analysis confirmed the proposal, as evidenced by the down-regulating level of Bcl-2 and the activation of caspase-3. Furthermore, the representative complexes Bi1, Bi4, Bi6, and Bi8 exhibited relatively lower inhibitory efficiency on human ovarian cancer cells (A2780) than on its cisplatin-resistant daughter cells (A2780/cis), thus demonstrating that such compounds are capable of circumventing the cisplatin-induced resistance. This investigation elucidated the excellent anticancer performance of C,O-coordinated bismuth(III) complexes and established the correlation between cytotoxic activity and coordination chemistry, which provides a practical basis for in-depth designing and developing bismuth-based chemotherapeutics.

Carbon-Iodine Polydiacetylene Nanofibers for Image-Guided Radiotherapy and Tumor-Microenvironment-Enhanced Radiosensitization.

Radiotherapy is a mainstay treatment used in clinics for locoregional therapy, although it still represents a great challenge to improve the sensitivity and accuracy of radiotherapy for tumors. Here, we report the conjugated polymer, polydiiododiacetylene (PIDA), with an iodine content of 84 wt %, as a highly effective computed tomography (CT) contrast agent and tumor microenvironment-responsive radiosensitizer. PIDA exhibited several key properties that contribute to the improvement of precision radiotherapy. The integrated PIDA nanofibers confined within the tumor envelope demonstrated amplified CT intensity and prolonged retention, providing an accurate calculation of dose distribution and precise radiation delivery for CT image-guided radiotherapy. Therefore, our strategy pioneers PIDA nanofibers as a bridge to cleverly connect a fiducial marker to guide accurate radiotherapy and a radiosensitizer to improve tumor sensitivity, thereby minimizing potential damage to surrounding tissues and facilitating on-demand therapeutic intervention in tumors.

[Determination of nine aromatic amines in water by cloud point extraction-gas chromatography-mass spectrometry].

Aromatic amines are a class of compounds bearing amino groups on their benzene rings; these compounds are important raw materials for the industrial production of rubber chemicals, pesticides, dyes, pharmaceuticals, photosensitive chemicals, and agricultural chemicals. Research has revealed that some aromatic amines teratogenetic, carcinogenic, and mutagenic properties. Given the high toxicity and potential harm caused by aromatic amines, monitoring their levels in water sources is critical. Aromatic amines are among the 14 strategic environmental pollutants blacklisted in China, and assessing their exposure levels is essential for protecting human health and the environment. At present, the standard method for detecting aromatic amines in water is liquid-liquid extraction-gas chromatography-mass spectrometry (LLE-GC-MS). However, this method has the disadvantages of large sample size requirement, complex operation, long analysis time, and high reagent consumption. In this study, instead of traditional LLE technology, cloud point extraction (CPE) technology was used in combination with GC-MS to establish an efficient, sensitive, and environment-friendly method for the detection of nine aromatic amines, namely, 2-chloramine, 3-chloramine, 4-chloramine, 2-nitroaniline, 3-nitroaniline, 4-nitroaniline, 1-naphthylamine, 2-naphthylamine, and 4-aminobenzene, in water. Triton X-114 was used as the extraction agent. The main experimental parameters were optimized using a single-factor optimization method. The aromatic amines in various water samples were quantitatively analyzed using GC-MS. The nine aromatic amines were separated on a DB-35 MS capillary column (30 m×0.25 mm×0.25 µm). The mass spectrometer was operated in selected ion monitoring (SIM) mode, and quantitative analysis was performed using the internal standard method. The results demonstrated that all nine aromatic amines could be completely separated within 16 min and had good linearities within accurate mass concentration ranges, with correlation coefficients (R2) greater than 0.998. The limits of detection (LODs) and quantification (LOQs) of these aromatic amines in water were 0.12-0.48 and 0.40-1.60 µg/L, respectively. The accuracy and precision of the method were assessed via the determination of aromatic amines in surface water of drinking water sources, offshore seawater, wastewater of the typical printing and dyeing industry at levels of 2.0 and 10.0 µg/L. The recoveries of the aromatic amines in surface water of drinking water sources were 81.1%-109.8%, with intra-day and inter-day relative standard deviations (RSDs) of 0.7%-5.2% (n=6) and 1.6%-6.2% (n=3), respectively. The recoveries of the aromatic amines in offshore seawater were 83.0%-115.8%, with intra-day RSDs (n=6) of 1.5%-8.6% and inter-day RSDs (n=3) of 2.4%-12.2%. The recoveries of the nine aromatic amines in wastewater of the typical printing and dyeing industry were 91.0%-120.0%, with intra-day RSDs (n=6) of 2.9%-12.9% and inter-day RSDs (n=3) of 2.5%-13.1%. The established method was used to detect nine aromatic amines in actual water samples. No aromatic amines were detected in the surface water of drinking water sources or offshore seawater samples. However, 2-chloramine, 4-chloramine, and 4-aminobenzene, which are frequently used in the printing and dyeing industry, were detected in the wastewater of the typical printing and dyeing industry samples. The proposed method offers the advantages of simple operation, high sensitivity, low cost, low organic reagent requirement, and good repeatability. Thus, this method provides reliable technical support for studying the residual status and environmental behavior of aromatic amines in water.

Nickel-Catalyzed C(sp3)-Sb Coupling of Chlorostibines with Unactivated Alkyl Chlorides and In Vitro Anticancer Activity of Products.

In this study, we present a nickel-catalyzed reductive C(sp3)-Sb coupling of unactivated alkyl chlorides with chlorostibines. This approach is highly versatile, tolerating various functional groups such as acetal, alkene, nitrile, amine, ester, silyl ether, thioether, and various heterocyclic compounds. Notably, the late-stage modification of bioactive molecules and the satisfactory anticancer activity against cancerous MDA-MB-231 also demonstrate the potential application.

Exercise-Induced miR-210 Promotes Cardiomyocyte Proliferation and Survival and Mediates Exercise-Induced Cardiac Protection against Ischemia/Reperfusion Injury.

Exercise can stimulate physiological cardiac growth and provide cardioprotection effect in ischemia/reperfusion (I/R) injury. MiR-210 is regulated in the adaptation process induced by exercise; however, its impact on exercise-induced physiological cardiac growth and its contribution to exercise-driven cardioprotection remain unclear. We investigated the role and mechanism of miR-210 in exercise-induced physiological cardiac growth and explored whether miR-210 contributes to exercise-induced protection in alleviating I/R injury. Here, we first observed that regular swimming exercise can markedly increase miR-210 levels in the heart and blood samples of rats and mice. Circulating miR-210 levels were also elevated after a programmed cardiac rehabilitation in patients that were diagnosed of coronary heart diseases. In 8-week swimming model in wild-type (WT) and miR-210 knockout (KO) rats, we demonstrated that miR-210 was not integral for exercise-induced cardiac hypertrophy but it did influence cardiomyocyte proliferative activity. In neonatal rat cardiomyocytes, miR-210 promoted cell proliferation and suppressed apoptosis while not altering cell size. Additionally, miR-210 promoted cardiomyocyte proliferation and survival in human embryonic stem cell-derived cardiomyocytes (hESC-CMs) and AC16 cell line, indicating its functional roles in human cardiomyocytes. We further identified miR-210 target genes, cyclin-dependent kinase 10 (CDK10) and ephrin-A3 (EFNA3), that regulate cardiomyocyte proliferation and apoptosis. Finally, miR-210 KO and WT rats were subjected to swimming exercise followed by I/R injury. We demonstrated that miR-210 crucially contributed to exercise-driven cardioprotection against I/R injury. In summary, this study elucidates the role of miR-210, an exercise-responsive miRNA, in promoting the proliferative activity of cardiomyocytes during physiological cardiac growth. Furthermore, miR-210 plays an essential role in mediating the protective effects of exercise against cardiac I/R injury. Our findings suggest exercise as a potent nonpharmaceutical intervention for inducing miR-210, which can alleviate I/R injury and promote cardioprotection.

miR-30d Attenuates Pulmonary Arterial Hypertension via Targeting MTDH and PDE5A and Modulates the Beneficial Effect of Sildenafil.

Pulmonary arterial hypertension (PAH) is associated with aberrant pulmonary vascular smooth muscle cell (PASMC) function and vascular remodeling. MiR-30d plays an important role in the pathogenesis of several cardiovascular disorders. However, the function of miR-30d in PAH progression remained unknown. Our study shows that circulating miR-30d level is significantly reduced in the plasma from PAH patients. In miR-30d transgenic (TG) rats, overexpressing miR-30d attenuates monocrotaline (MCT)-induced pulmonary hypertension (PH) and pulmonary vascular remodeling. Increasing miR-30d also inhibits platelet-derived growth factor-bb (PDGF-bb)-induced proliferation and migration of human PASMC. Metadherin (MTDH) and phosphodiesterase 5A (PDE5A) are identified as direct target genes of miR-30d. Meanwhile, nuclear respiratory factor 1 (NRF1) acts as a positive upstream regulator of miR-30d. Using miR-30d knockout (KO) rats treated with sildenafil, a PDE5A inhibitor that is used in clinical PAH therapies, it is further found that suppressing miR-30d partially attenuates the beneficial effect of sildenafil against MCT-induced PH and vascular remodeling. The present study shows a protective effect of miR-30d against PAH and pulmonary vascular remodeling through targeting MTDH and PDE5A and reveals that miR-30d modulates the beneficial effect of sildenafil in treating PAH. MiR-30d should be a prospective target to treat PAH and pulmonary vascular remodeling.

[Determination of 14 aniline and benzidine compounds in soil by gas chromatography-mass spectrometry].

The complex matrix of soil samples and low extraction efficiency of aniline compounds limit many methods developed for detecting aniline and benzidine compounds in soil. In this study, a rapid and sensitive method based on gas chromatography-mass spectrometry was developed for the simultaneous determination of 14 aniline and benzidine compounds in soil. The collected soil samples were sealed with 5% sodium sulfite solution and refrigerated to inhibit the oxidation of the target compounds for up to 7 d. The extraction efficiencies of accelerated solvent extraction and oscillating dispersion extraction were compared, and the recovery of accelerated solvent extraction was found to be unsuitable. Hence, three-phase oscillating dispersion extraction was adopted. A certain amount of alkaline aqueous solution was added to the test system during extraction to improve the extraction efficiency because aniline and benzidine compounds are weakly alkaline substances. When the pH of the extracted water phase was greater than 12, relatively good recoveries were obtained. Next, a mixed solvent of ethyl acetate-methylene chloride (1∶4, v/v) was added to extract the target compounds via oscillation for 20 min. The solid phase was discarded via centrifugation, and the aqueous and organic phases were transferred to a liquid separation funnel for further separation. Finally, the organic phase was retained. This pretreatment process prevents the co-extraction of acidic compounds or other impurities, thereby enhancing the purification ability of the method. Solid phase extraction (SPE) is generally recommended for soil extraction and purification. A preliminary test showed that compared with other columns, the Florisil SPE column could better retain the target substances and exhibited higher elution efficiency. After purification, the organic phase was concentrated to 1 mL using a nitrogen blower. The analytes were analyzed by gas chromatography-mass spectrometry using a capillary column (DB-35MS, 30 m×0.25 mm×0.25 µm). The temperature program was optimized to separate the target compounds at the baseline. Specifically, the initial oven temperature was set to 60 ℃, held for 2 min, increased to 130 ℃ at a rate of 5 ℃/min, increased to 300 ℃ at a rate of 30 ℃/min, and held for 4 min. The injector and ion source temperatures were 250 and 300 ℃, respectively. Aniline-d5 and acenaphthene-d10 were used as the internal standards for quantification. The effects of antioxidant addition, extraction solvent type, salting out, and other factors on extraction efficiency were investigated. The results showed that the method performed well under the optimized experimental conditions when actual soils were used as real sample matrices. The accuracy and precision of the proposed method were verified. A total of 14 aniline and benzidine compounds demonstrated good linearities in the range of 0.5-100 mg/L. The method detection limits (MDLs) ranged from 0.02 to 0.07 mg/kg, and the limits of quantification (LOQs) ranged from 0.08 to 0.28 mg/kg. The target compounds were spiked at contents of 1 and 10 mg/kg. The spiked recoveries of the 14 targets in actual soils were 62.9%-101%, and the relative standard deviations (RSDs) of six precision tests were 3.8%-10.3%. The proposed method effectively inhibited the oxidation of aniline and benzidine compounds during extraction, and the target compounds exhibited high recoveries and good stabilities in the presence of three phases. Moreover, the operating procedure was simple and easy to implement. The proposed method was applied to the soil collected from an industrial enterprise in Jiangsu province that was suspected to be contaminated with aniline, and two aniline compounds were detected. The developed method requires a small sample size, and the preservation step is simple and effective. In addition, it can be applied to various types of actual soils. The method meets the requirements of current soil pollution risk control standards for aniline and benzidine compounds in soils.

Sodium alginate-based composite microspheres for controlled release of pesticides and reduction of adverse effects of copper in agricultural soils.

Excessive copper (Cu) concentrations pose significant health risks to both plants and humans. In this study, sodium alginate (SA)-gelatin (GEL)-polyvinyl pyrrolidone (PVP)- embedded dinotefuran (DIN) microspheres were prepared using spray-drying technology. The loading content and encapsulation efficiency of optimal microspheres determined by physical modifications were 19.77% and 99.32%, respectively. In addition, the microspheres showed variable stimuli-responsive controlled release capacities in different temperatures and types of soil, as well as showed better control efficiency of larvae of Protaetia brevitarsis at pesticide application in the early stage, with the potential ability to control pest outbreaks at high temperatures. In addition, blank microspheres improved the growth and physiological activity of cucumber seedlings, reduced copper content in leaves, increased soil nutrient content, and prevented soil acidification. Further, the use of blank microspheres increased the relative abundance of soil beneficial functional bacteria communities, which mediate heavy metal (HM) immobilization/tolerance and promote plant growth. Redundancy analysis (RDA) and Spearman correlation analysis showed that these beneficial functional bacteria were mainly positively correlated with soil EC, A-N, and N-N. In summary, this study showed that the technique of combining physically modified carrier materials with pesticides has the potential to reduce Cu contamination in the surrounding agricultural soil during pesticide application, thereby reducing Cu uptake by crops.

Effect of sodium alginate-gelatin-polyvinyl pyrrolidone microspheres on cucumber plants, soil, and microbial communities under lead stress.

Lead is highly persistent and toxic in soil, hindering plant growth. Microspheres are a novel, functional, and slow-release preparation commonly used for controlled release of agricultural chemicals. However, their application in the remediation of Pb-contaminated soil has not been studied; furthermore, the remediation mechanism involved has not been systematically assessed. Herein, we evaluated the Pb stress mitigation ability of sodium alginate-gelatin-polyvinyl pyrrolidone composite microspheres. Microspheres effectively attenuated the Pb toxic effect on cucumber seedlings. Furthermore, they boosted cucumber growth, increased peroxidase activity, and chlorophyll content, while reducing malondialdehyde content in leaves. Microspheres promoted Pb enrichment in cucumber, especially in roots (about 4.5 times). They also improved soil physicochemical properties, promoted enzyme activity, and increased soil available Pb concentration in the short term. In addition, microspheres selectively enriched functional (heavy metal-tolerating and plant growth promoting) bacteria to adapt to and resist Pb stress by improving soil properties and nutrients. These results indicated that even a small amount (0.025-0.3 %) of microspheres can significantly reduce the adverse effects of Pb on plants, soil, and bacterial communities. Composite microspheres have shown great value in Pb remediation, and their application potential in phytoremediation is also worth evaluating to expand the application.

Danlou Tablet Protects Against Myocardial Infarction Through Promoting eNOS-Dependent Endothelial Protection and Angiogenesis.

Myocardial infarction (MI) is one of the leading causes of death worldwide. Danlou tablet (Dan) is an effective traditional Chinese medicine for cardiac protection, although the underlying mechanism was not fully understood. In this study, we used a murine MI model and demonstrated that Dan administration effectively attenuated myocardial apoptosis, cardiac remodeling, and heart failure post MI. Dan increased CD31-positive capillaries in MI hearts, and reduced the apoptosis and oxidative stress in human umbilical vein endothelial cells after oxygen-glucose deprivation stress, simultaneously with the activated HIF-1α/VEGFA/eNOS signaling. Moreover, inhibition of eNOS by L-NAME attenuated Dan-induced protection against MI, and abolished its effect in promoting angiogenesis and reducing endothelial apoptosis and oxidative stress. Collectively, Dan is beneficial to promote eNOS-dependent endothelial protection and angiogenesis thus protecting against MI. A deep understanding of Dan-induced protection might help promote clinical usage of Dan in MI treatment.

Danlou Tablet Protects Against Cardiac Remodeling and Dysfunction after Myocardial Ischemia/Reperfusion Injury through Activating AKT/FoxO3a Pathway.

Myocardial ischemia/reperfusion injury (I/RI) and ventricular remodeling are the critical pathological basis of heart failure. Danlou tablet (Dan) is a kind of Chinese patent medicine used in angina pectoris treatment in China. However, it remains unclear whether and how Dan could protect against cardiac remodeling after myocardial I/RI. In this study, both preventive and therapeutic administration of Dan attenuated ventricular remodeling and cardiac dysfunction at 3 weeks after myocardial I/RI. Dan inhibited Bax/Bcl2 ratio and Caspase3 cleavage in heart tissues and also inhibited apoptosis of human AC16 cells and neonatal rat cardiomyocytes stressed by oxygen and glucose deprivation/reperfusion. Mechanistically, Dan inhibited myocardial apoptosis through phosphorylating AKT and FoxO3a, thereby inhibiting downstream BIM and PUMA expressions. Collectively, these results demonstrate that Dan treatment is effective to protect against cardiac remodeling and dysfunction after myocardial I/RI and provide theoretical basis for its cardioprotection and clinical application in treating ischemic cardiac diseases.

Targeted Computed Tomography Visualization and Healing of Inflammatory Bowel Disease by Orally Delivered Bacterial-Flagella-Inspired Polydiiododiacetylene Nanofibers.

Accurate diagnosis and timely therapeutic intervention of inflammatory bowel disease (IBD) is essential in preventing the progression of the disease, although it still represents an insurmountable challenge. Here we report the design of bacterial-flagella-inspired polydiiododiacetylene (PIDA) nanofibers and its performance in targeted computed tomography (CT) imaging and on-demand therapeutic intervention of IBD. With a morphology mimicking bacterial flagella, PIDA nanofibers attach on the mucus layer of the gastrointestinal (GI) tract after oral administration, evenly distributing on the GI surface to portray the GI lining under CT scan within 2 h. PIDA can retain for a longer time in the damaged mucosa at the inflamed lesions than in normal GI tissues to enable the targeted CT visualization of IBD. PIDA also scavenges reactive oxygen species and ameliorates gut dysbiosis attributed to its iodine-substituted polydiacetylene structure, so that the enriched PIDA nanofibers at the targeted IBD lesions can alleviate the inflammation while maintaining the gut microbiota homeostasis, thus promoting the rebalance of GI microenvironment and the mucosal healing.