Metabolic syndrome facilitates histopathological changes and the risk of postoperative recurrence in chronic rhinosinusitis with nasal polyps.

BACKGROUND: The relationship between metabolic syndrome (MS) and chronic rhinosinusitis with nasal polyps (CRSwNP) remains unclear. This study aimed to examine the effects of MS on histopathological features and postoperative recurrence in patients with CRSwNP. METHODS: We recruited 529 patients with CRSwNP who underwent functional endoscopic sinus surgery. They were divided into MS and non-MS groups and followed up for 2 years to evaluate postoperative recurrence. Clinical characteristics, histopathological features, the immunoactivity of signature cytokines, and the risk of postoperative recurrence were compared between the two groups. RESULTS: In total, 490 patients with CRSwNP were included in the study, 145 of whom experienced postoperative recurrence. The recurrence rate, tissue eosinophil count and percentage, and expression levels of IL-5 and IL-17A were significantly higher in the MS group compared to the non-MS group. Furthermore, within the MS group, patients who experienced recurrence exhibited higher tissue eosinophil counts and IL-5 and IL-17A levels than those in the non-MS group. Notably, the eosinophil count and IL-5 and IL-17A levels were higher in tissues collected during revision surgery than in those collected during primary surgery, particularly in patients with MS. Binary logistic regression analysis and Kaplan-Meier survival curves consistently indicated that MS independently increased the risk of postoperative recurrence in patients with CRSwNP. Furthermore, the risk increased with the number of MS components presented. CONCLUSION: MS promoted tissue eosinophil infiltration, and IL-5 and IL-17A expression, and increased the risk of postoperative recurrence in patients with CRSwNP. MS was identified as an independent risk factor for postoperative recurrence, and the risk increased with an increase in the number of MS components.

A Novel EYA1 Mutation Causing Alternative RNA Splicing in a Chinese Family With Branchio-Oto Syndrome: Implications for Molecular Diagnosis and Clinical Application.

OBJECTIVES: Branchio-oto syndrome (BOS) primarily manifests as hearing loss, preauricular pits, and branchial defects. EYA1 is the most common pathogenic gene, and splicing mutations account for a substantial proportion of cases. However, few studies have addressed the structural changes in the protein caused by splicing mutations and potential pathogenic factors, and several studies have shown that middle-ear surgery has limited effectiveness in improving hearing in these patients. BOS has also been relatively infrequently reported in the Chinese population. This study explored the genetic etiology in the family of a proband with BOS and provided clinical treatment to improve the patient's hearing. METHODS: We collected detailed clinical features and peripheral blood samples from the patients and unaffected individuals within the family. Pathogenic mutations were identified by whole-exome sequencing and cosegregation analysis and classified according to the American College of Medical Genetics and Genomics guidelines. Alternative splicing was verified through a minigene assay. The predicted three-dimensional protein structure and biochemical experiments were used to investigate the pathogenicity of the mutation. The proband underwent middle-ear surgery and was followed up at 1 month and 6 months postoperatively to monitor auditory improvement. RESULTS: A novel heterozygous EYA1 splicing variant (c.1050+4 A>C) was identified and classified as pathogenic (PVS1(RNA), PM2, PP1). Skipping of exon 11 of the EYA1 pre-mRNA was confirmed using a minigene assay. This mutation may impair EYA1-SIX1 interactions, as shown by an immunoprecipitation assay. The EYA1-Mut protein exhibited cellular mislocalization and decreased protein expression in cytological experiments. Middle-ear surgery significantly improved hearing loss caused by bone-conduction abnormalities in the proband. CONCLUSION: We reported a novel splicing variant of EYA1 in a Chinese family with BOS and revealed the potential molecular pathogenic mechanism. The significant hearing improvement observed in the proband after middle-ear surgery provides a reference for auditory rehabilitation in similar patients.

RNA binding motif protein 45-mediated phosphorylation enhances protein stability of ASCT2 to promote hepatocellular carcinoma progression.

Targeting metabolic remodeling represents a potentially promising strategy for hepatocellular carcinoma (HCC) therapy. In-depth understanding on the regulation of the glutamine transporter alanine-serine-cysteine transporter 2 (ASCT2) contributes to the development of novel promising therapeutics. As a developmentally regulated RNA binding protein, RBM45 is capable to shuttle between nucleus and cytoplasm, and directly interacts with proteins. By bioinformatics analysis, we screened out that RBM45 was elevated in the HCC patient specimens and positively correlated with poor prognosis. RBM45 promoted cell proliferation, boosted xenograft tumorigenicity and accelerated HCC progression. Using untargeted metabolomics, it was found that RBM45 interfered with glutamine metabolism. Further results demonstrated that RBM45 positively associated with ASCT2 in human and mouse specimens. Moreover, RBM45 enhanced ASCT2 protein stability by counteracting autophagy-independent lysosomal degradation. Significantly, wild-type ASCT2, instead of phospho-defective mutants, rescued siRBM45-suppressed HCC cell proliferation. Using molecular docking approaches, we found AG-221, a mutant isocitrate dehydrogenase 2 (mIDH2) inhibitor for acute myeloid leukemia therapy, pharmacologically perturbed RBM45-ASCT2 interaction, decreased ASCT2 stability and suppressed HCC progression. These findings provide evidence that RBM45 plays a crucial role in HCC progression via interacting with and counteracting the degradation of ASCT2. Our findings suggest a novel alternative structural sites for the design of ASCT2 inhibitors and the agents interfering with RBM45-ASCT2 interaction may be a potential direction for HCC drug development.

Effects of gene silencing of indoleamine 2,3-dioxygenase 1 combined with rosmarinic acid on tumor immune microenvironment in H22 tumor-bearing mice.

Rosmarinic acid (RA) is a natural polyphenolic compound with several pharmacological activities, including immunomodulation and anti-tumor effect. Indoleamine 2,3-dioxygenase-1 (IDO1), the rate-limiting enzyme that metabolizes tryptophan into kynurenine, is an important negative immune regulator. This study aimed to explore the effect of combined action of IDO1 gene silencing and RA on tumor immune microenvironment. H22 tumor-bearing mice were treated with combination therapy with RA and IDO1-shRNA. The percentages and apoptosis of T-cells and subsets of splenic regulatory T-cells (Tregs) were detected by flow cytometry. Levels of tumor necrosis factor (TNF-α), Interferon-γ (IFN-γ), interleukin-2 (IL-2) and interleukin-10 (IL-10) were measured by enzyme linked immunosorbent assay (ELISA). Treatment with RA + IDO1-shRNA significantly increased the percentage of CD4+ T cells, ratio of CD4+/CD8+ and the levels of IFN-γ and IL-2, while decreased CD8+ apoptosis, the proportion of splenic Tregs and the levels of TNF-α and IL-10. The present study demonstrated that combination therapy with RA and IDO1-shRNA had anti-tumor effects on HCC. The mechanism might be related to regulating immune response and immunocytokines, as well as alleviating immunosuppression induced by Tregs in the tumor immune microenvironment.

Overcoming ABCB1-mediated multidrug resistance by transcription factor BHLHE40.

Multidrug resistance (MDR) hinders treatment efficacy in cancer therapy. One typical mechanism contributing to MDR is the overexpression of permeability-glycoprotein (P-gp) encoded by ATP-binding cassette subfamily B member 1 (ABCB1). Basic helix-loop-helix family member e40 (BHLHE40) is a well-known transcription factor that has pleiotropic effects including the regulation of cancer-related processes. However, whether BHLHE40 regulates MDR is still unknown. Chromatin immunoprecipitation-seq study revealed BHLHE40 occupancy in the promoter of ABCB1 gene. Adriamycin (ADM)-resistant human chronic myeloid leukemia cells (K562/A) and human breast cancer cells (MCF-7/A) were established. BHLHE40 expression was downregulated in the ADM-resistant cell lines. Overexpression of BHLHE40 resensitized resistant cells to ADM, promoted cell apoptosis in vitro and suppressed tumor growth in vivo, whereas BHLHE40 knockdown induced resistance to ADM in parental cells. Moreover, we found that BHLHE40 regulated drug resistance by directly binding to the ABCB1 promoter (-1605 to -1597) and inactivating its transcription. In consistence, the expression of BHLHE40 was negatively correlated with ABCB1 in various cancer cells, while positively with cancer cell chemosensitivity and better prognosis of patients with breast cancer. The study reveals the role of BHLHE40 as a transcriptional suppressor on the expression of ABCB1, major ABC transporter in chemoresistance. The findings extend the function of BHLHE40 in tumor progression and provides a novel mechanism for the reversal of multidrug resistance.

A comprehensive genotype-phenotype evaluation of eight Chinese probands with Waardenburg syndrome.

BACKGROUND: Waardenburg syndrome (WS) is the most common form of syndromic deafness with phenotypic and genetic heterogeneity in the Chinese population. This study aimed to clarify the clinical characteristics and the genetic cause in eight Chinese WS families (including three familial and five sporadic cases). Further genotype-phenotype relationships were also investigated. METHODS: All probands underwent screening for the known WS-related genes including PAX3, SOX10, MITF, EDNRB, EDN3, and SNAI2 using next-generation sequencing to identify disease-causing genes. Further validation using Sanger sequencing was performed. Relevant findings for the associated genotype-phenotype from previous literature were retrospectively analyzed. RESULT: Disease-causing variants were detected in all eight probands by molecular genetic analysis of the WS genes (SOX10(NM_006941.4): c.544_557del, c.553 C > T, c.762delA, c.336G > A; MITF(NM_000248.3): c.626 A > T; PAX3(NM_181459.4): c.838delG, c.452-2 A > G, c.214 A > G). Six mutations (SOX10:c.553 C > T, c.544_557del, c.762delA; PAX3: c.838delG, c.214 A > G; MITF:c.626 A > T) were first reported. Clinical evaluation revealed prominent phenotypic variability in these WS patients. Twelve WS1 cases and five WS2 cases were diagnosed in total. Two probands with SOX10 mutations developed progressive changes in iris color with age, returning from pale blue at birth to normal tan. Additionally, one proband had a renal malformation (horseshoe kidneys).All cases were first described as WS cases. Congenital inner ear malformations were more common, and semicircular malformations were exclusively observed in probands with SOX10 mutations. Unilateral hearing loss occurred more often in cases with PAX3 mutations. CONCLUSION: Our findings helped illuminate the phenotypic and genotypic spectrum of WS in Chinese populations and could contribute to better genetic counseling of WS.

Molecular Link in Flavonoid and Amino Acid Biosynthesis Contributes to the Flavor of Changqing Tea in Different Seasons.

The present study was aimed to elucidate the flavor formation mechanism of Changqing tea. High-performance liquid chromatography (HPLC) analysis showed that the total catechins of Changqing tea was 65-160 mg/g, with 16-34 mg/g non-galloyated catechins and 49-126 mg/g galloylated catechins. Tea polyphenols and free amino acids account for 286-312 mg/g and 35-89 mg/g, respectively. Transcriptome of Changqing tea during different seasons revealed 316, 130 and 12 DEGs in comparisons of spring vs. autumn, spring vs. summer, and summer vs. autumn, respectively. Compared to spring, the genes involved in flavonoid biosynthesis and bitter imparted amino acids were up-regulated in summer and autumn. Metabolome analysis was conducted by using HPLC-MS; the result indicated that umami and kokumi contributing amino acids were decreased in summer and autumn compared with spring. It could be concluded that the coordination of flavonoid biosynthesis and amino acids biosynthesis resulted in the special flavor of Changqing tea.

Metabolic dysregulation and emerging therapeutical targets for hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is an aggressive human cancer with increasing incidence worldwide. Multiple efforts have been made to explore pharmaceutical therapies to treat HCC, such as targeted tyrosine kinase inhibitors, immune based therapies and combination of chemotherapy. However, limitations exist in current strategies including chemoresistance for instance. Tumor initiation and progression is driven by reprogramming of metabolism, in particular during HCC development. Recently, metabolic associated fatty liver disease (MAFLD), a reappraisal of new nomenclature for non-alcoholic fatty liver disease (NAFLD), indicates growing appreciation of metabolism in the pathogenesis of liver disease, including HCC, thereby suggesting new strategies by targeting abnormal metabolism for HCC treatment. In this review, we introduce directions by highlighting the metabolic targets in glucose, fatty acid, amino acid and glutamine metabolism, which are suitable for HCC pharmaceutical intervention. We also summarize and discuss current pharmaceutical agents and studies targeting deregulated metabolism during HCC treatment. Furthermore, opportunities and challenges in the discovery and development of HCC therapy targeting metabolism are discussed.

Phosphorylation-Induced Ubiquitination and Degradation of PXR through CDK2-TRIM21 Axis.

Pregnane X receptor (PXR) is a member of the nuclear receptor superfamily that is activated by a variety of endogenous metabolites or xenobiotics. Its downstream target genes are involved in metabolism, inflammation and processes closely related to cancer. However, the stability regulation of PXR protein resulting from post-translational modification is still largely undefined. In the present study, primary mouse hepatocytes, hepatoma HepG2 cells and HEK 293T cells were used to investigate gene expression and protein interactions. The role of kinases was evaluated by RNA interference and overexpression constructs with or without PXR phosphorylation site mutations. The activity of CYP3A4 and P-gp was determined by enzymatic and substrate accumulation assays. It was found that E3 ubiquitin ligase TRIM21 mediates the ubiquitination and degradation of PXR and plays an important role in regulating the activity of PXR. On this basis, PXR phosphorylation-associated kinases were evaluated regarding regulation of the stability of PXR. We found cyclin dependent kinase 2 (CDK2) exclusively phosphorylates PXR at Ser350, promotes its disassociation with Hsp90/DNAJC7, and leads to subsequent TRIM21-mediated PXR ubiquitination and degradation. As well-known CDK inhibitors, dinaciclib and kenpaullone stabilize PXR and result in elevated expression and activity of PXR-targeted DMETs, including carboxylesterases, CYP3A4 and P-gp. The suppressed degradation of PXR by CDK2 inhibitors denotes dinaciclib-induced promotion of PXR-targeted genes. The findings of CDK2-mediated PXR degradation indicate a wide range of potential drug-drug interactions during clinical cancer therapy using CDK inhibitors and imply an alternative direction for the development of novel PXR antagonists.

Response of extracellular and intracellular alkaline phosphatase in Microcystis aeruginosa to organic phosphorus.

Cyanobacterial blooms caused by Microcystis have become a menace to public health and water quality in the global freshwater ecosystem. Alkaline phosphatases (APases) produced by microorganisms play an important role in the mineralization of dissolved organic phosphorus (DOP) into orthophosphate (Pi) to promote cyanobacterial blooms. However, the response of extracellular and intracellular alkaline phosphatase activity (APA) of Microcystis to different DOP sources is poorly understood. In this study, we compared the growth of M. aeruginosa on two DOP substrates (ß-glycerol-phosphate (ß-GP) and lecithin (LEC)) and monitored the changes of P fractions and the extra- and intracellular APA under different P sources and concentrations. M. aeruginosa can utilize both ß-GP and LEC to sustain its growth, and the bioavailability of LEC was greater than ß-GP. For the ß-GP treatment, there was no significant difference in the algal growth at different concentrations (P > 0.05), while the algal growth in the LEC treatment groups was significantly affected by concentrations (P < 0.05). The results showed that intracellular APA of M. aeruginosa could be detected in all DOP treatment groups and generally higher than extracellular APA. In addition, the intracellular APA per cell increased first and then decreased in all DOP treatment groups. Compared with the ß-GP treatment, M. aeruginosa in the LEC groups could secret more extracellular APA.

Suspended particles phoD alkaline phosphatase gene diversity in large shallow eutrophic Lake Taihu.

The bacterial phoD gene encodes alkaline phosphatase plays an important role in the release of bioavailable inorganic phosphorus (P) from organic P in environmental systems. However, phoD gene diversity in suspended particles in shallow freshwater lakes is poorly understood. In this study, we explored the potential relationship between environmental factors and phoD phosphatase gene in suspended particles in different ecosystem types (lake zones) in Lake Taihu, a large shallow eutrophic lake in China. Quantitative PCR and high-throughput sequencing were used to analyze phoD gene abundance and the phoD-harboring bacterial community composition. Our results indicate that the distribution of phoD gene abundance in suspended particles had a high spatiotemporal heterogeneity. The phoD gene abundance in each lake zone decreased significantly from June to September. The dominant phoD-harboring phylum in all samples was Actinobacteria, followed by Proteobacteria, Cyanobacteria and Gemmatimonadetes. The first predominant phoD-harboring genera varied among samples, but most of them belonged to phylum Actinobacteria. Driven by different environmental factors, the phoD-harboring bacterial community structure varied with sampling month and ecosystem type. Nitrate and ammonia nitrogen were the main environmental drivers of phoD-harboring bacterial community in suspended particles in the river mouth zone, while water pH and dissolved oxygen were important factors for the algae-dominated, macrophyte-dominated and central lake zones.

Intradermal delivery of STAT3 siRNA to treat melanoma via dissolving microneedles.

Hyperactivity of signal transducer and activity of transcription 3 (STAT3) plays a crucial role in melanoma invasion and metastasis. Gene therapy applying siRNA targeting STAT3 is a potential therapeutic strategy for melanoma. In this article, we first fabricated safe and novel dissolving microneedles (MNs) for topical application of STAT3 siRNA to enhance the skin penetration of siRNA and used polyethylenimine (PEI, 25 kDa) as carrier to improve cellular uptake of siRNA. The results showed that MNs can effectively penetrate skin and rapidly dissolve in the skin. In vitro B16F10 cell experiments presented that STAT3 siRNA PEI complex can enhance cellular uptake and transfection of siRNA, correspondingly enhance gene silencing efficiency and inhibit tumor cells growth. In vivo experiments indicated that topical application of STAT3 siRNA PEI complex delivered by dissolving MNs into skin can effectively suppress the development of melanoma through silencing STAT3 gene, and the inhibition effect is dose-dependent. STAT3 siRNA delivery via dissolving MNs is a promising approach for skin melanoma treatment with targeting inhibition efficacy and minimal adverse effects.

A technology for strongly improving methane production from rice straw: freeze-thaw pretreatment.

Overcoming the complex three dimensional structure of biomass is a major challenge in enhancing anaerobic digestion (AD) efficacy. Freeze-thaw pretreatment was proposed herein in order to improve methane production from rice straw. The effect was notable: average methane content for group-A (-4 °C) and -B (-20 °C) were A1 (-4 °C, 12 h): 40.0%, A2 (-4 °C, 24 h): 40.5%, A3 (-4 °C, 48 h): 42.2%; B1 (-20 °C, 12 h): 44.2%, B2 (-20 °C, 24 h): 45.7%, B3 (-20 °C, 48 h): 46.0%, the increases were 88.8-99.1% and 108.8-117.2%, respectively, compared with control (CK) (21.2%). Total methane production for group-A and -B were A1: 22.8 mL g-1 TS, A2: 24.7 mL g-1 TS, A3: 27.8 mL g-1 TS; B1: 29.9 mL g-1 TS, B2: 31.3 mL g-1 TS, B3: 32.0 mL g-1 TS, compared with CK (7.6 mL g-1 TS), the increases were 200.0-265.8%, 293.4-321.1%, respectively. The technical digestion time (T 80) was shortened by 8 days. Therefore, the maximum methane production was obtained under conditions of -20 °C and 48 h. This study proposed an efficient pretreatment method that broadens the horizon of improving biomass conversion into bioenergy.

Dual roles of TPGS based microemulsion for tacrolimus: Enhancing the percutaneous delivery and anti-psoriatic efficacy.

In this study, we demonstrate for the first time the dual roles of Tocopheryl Polyethylene Glycol 1000 Succinate (TPGS) based microemulsion (ME) for tacrolimus (TAC) to enhance TAC percutaneous delivery and anti-psoriatic efficacy. The ME formulation was developed and optimized based on pseudo-ternary phase diagrams combined with in vitro permeation. The result of Fourier transform infrared spectroscopy (FTIR) demonstrated that TAC was completely solubilized in the TPGS-ME. In vitro permeation studies showed that TPGS-ME enhanced TAC permeation through and into the skin, and the enhanced deposition of TAC in the normal or psoriatic skin was further confirmed in vivo. The cellular uptake performed with HaCaT cells presented more pronounced uptake of TPGS-ME. Topical TAC-TPGS-ME treated imiquimod-induced psoriasis in mice more efficaciously than the commercial formulation of TAC (Protopic®), which is consistent with the enhanced TAC levels by TAC-TPGS-ME in the psoriatic skin. Haematoxylin and Eosin (HE) staining revealed that TAC-TPGS-ME significantly diminished the severity of the psoriasis-like skin inflammation. Moreover, TPGS-ME vehicle exhibited a moderate anti-inflammatory activity, which indicated that TPGS acted as a potential adjuvant in the TAC anti-psoriasis process, and the synergism was identified in anti-proliferation against HaCaT cells. The colloidal nano-carrier combined ME with TPGS is a potential approach for percutaneous delivery of TAC, which exploited both virtues of ME and TPGS to obtain the synergetic effects of enhanced permeability and anti-psoriatic efficacy.

Effects of nanoparticles with hydrotropic nicotinamide on tacrolimus: permeability through psoriatic skin and antipsoriatic and antiproliferative activities.

The hybrid system based on nanoparticles (NPs) self-assembled by the conjugations of hyaluronic acid with cholesterol (HA-Chol NPs) combined with nicotinamide (NIC) for tacrolimus (FK506), ie, FK506 NPs-NIC, has been confirmed to exhibit a significant synergistic effect on FK506 permeation through and into intact skin; thus, it may be a promising approach for FK506 to effectively treat skin diseases. The aim of this study was to evaluate its potential for the treatment of psoriasis. In vitro permeation through the psoriatic skin was carried out, and the results revealed that the combination of NPs with NIC exhibited a significant synergistic effect on FK506 deposition within the psoriatic skin (3.40±0.67 µg/cm2) and penetration through the psoriatic skin (30.86±9.66 µg/cm2). The antipsoriatic activity of FK506 NPs-NIC was evaluated through the treatment for imiquimod (IMQ)-induced psoriasis. The psoriasis area and severity index (PASI) score demonstrated that FK506 HA-Chol NPs-NIC exerted the effect on ameliorating the skin lesions comparable to clobetasol propionate (a positive drug for psoriasis) and superior to commercial FK506 ointment (Protopic®), and the histological study showed that it presented a synergistic effect on antipsoriasis after FK506 incorporation into NPs combined with NIC hydrotropic system, which might ultimately increase the therapeutic effect and minimize the systemic side effects by reducing the overall dose of FK506. RAW 264.7 cell uptake presented the enhancement of drugs delivered into cells by HA-Chol NPs-NIC. The antiproliferative activity on HaCaT cells identified that FK506 HA-Chol NPs-NIC exhibited significant inhibiting effects on HaCaT proliferation. The results support that the combination of HA-Chol NPs with NIC is a promising approach for FK506 for the treatment of psoriasis.

Combination of hydrotropic nicotinamide with nanoparticles for enhancing tacrolimus percutaneous delivery.

Tacrolimus (FK506), an effective immunosuppressant for treating inflammatory skin diseases, hardly penetrates into and through the skin owing to its high hydrophobicity and molecular weight. The aim of this study was to develop a hybrid system based on nicotinamide (NIC) and nanoparticles (NPs) encapsulating FK506, such as FK506-NPs-NIC, for facilitating percutaneous delivery, which exploited virtues of both NIC and NPs to obtain the synergetic effect. Solubility and percutaneous permeation studies were carried out. The results showed that NIC could increase the solubility and permeability of FK506 and that 20% (w/v) NIC presented higher FK506 permeability and was thus chosen as the hydrotropic solution to solubilize FK506 and prepare FK506-NPs-NIC. Hyaluronic acid (HA) was chemically conjugated with cholesterol (Chol) to obtain amphiphilic conjugate of HA-Chol, which self-assembled NPs in 20% NIC solution containing FK506. The particle size, zeta potential, and morphology of NPs were characterized. The encapsulation efficiency and in vitro percutaneous permeation of NPs were evaluated in the presence and absence of NIC. The results demonstrated that hydrotropic solubilizing FK506 was readily encapsulated into NPs with a higher encapsulation efficiency of 79.2%±4.2%, and the combination of NPs with NIC exhibited a significantly synergistic effect on FK506 deposition within the skin (2.39±0.53 µg/cm(2)) and penetration through the skin (13.38±2.26 µg/cm(2)). The effect of the combination of NPs with NIC on drug permeation was further visualized by confocal laser scanning microscope through in vivo permeation studies, and the results confirmed that NPs-NIC synergistically enhanced the permeation of the drug into the skin. The cellular uptake performed in HaCaT cells presented a promoting effect of NPs on cellular uptake. These overall results demonstrated that HA-Chol-NPs-NIC can synergistically improve the percutaneous delivery of FK506, and it is a novel potential strategy based on a nano-sized carrier for FK506 to treat skin diseases.

Hyaluronic acid-coated niosomes facilitate tacrolimus ocular delivery: Mucoadhesion, precorneal retention, aqueous humor pharmacokinetics, and transcorneal permeability.

Tacrolimus (FK506) was used to prevent corneal allograft rejection in patients who were resistant to steroids and cyclosporine. However, the formulation for FK506 ocular delivery remained a challenge due to the drug's high hydrophobicity, high molecular weight, and eye's physiological and anatomical constraints. The aim of this project is to develop an ocular delivery system for FK506 based on a combined strategy of niosomes and mucoadhesive hyaluronic acid (HA), i.e., FK506HA-coated niosomes, which exploits virtues of both niosomes and HA to synergistically improve ophthalmic bioavailability. The FK506HA-coated niosomes were characterized with particle size, zeta potential, and rheology behavior. Mucoadhesion of FK506HA-coated niosomes to mucin was investigated through surface plasmon resonance in comparison with non-coated niosomes and HA solution. The results showed that niosomes possessed adhesion to mucin, and HA coating enhanced the adhesion. The in vivo precorneal retention was evaluated in rabbit, and the results showed that HA-coated niosomes prolonged the residence of FK506 significantly in comparison with non-coated niosomes or suspension. Aqueous humor pharmacokinetics test showed that area under curve of HA-coated niosomes was 2.3-fold and 1.2-fold as that of suspension and non-coated niosomes, respectively. Moreover, the synergetic corneal permeability enhancement of the hybrid delivery system on FK506 was visualized and confirmed by confocal laser scanning microscope. Overall, the results indicated that the hybrid system facilitated FK506 ocular delivery on mucoadhesion, precorneal retention, aqueous humor pharmacokinetics and transcorneal permeability. Therefore, HA-coated niosomes may be a promising approach for ocular targeting delivery of FK506.

In vivo cancer targeting and fluorescence-CT dual-mode imaging with nanoprobes based on silver sulfide quantum dots and iodinated oil.

In this article, a fluorescence-CT dual-mode nanoprobe is successfully synthesized by making use of distearoylphosphatidylethanolamine-poly(ethylene glycol)-folate (DSPE-PEG2000-FA) and other amphiphilic molecules to coat silver sulfide (Ag2S) quantum dots (QDs) and iodinated oil simultaneously. In vitro experiments show that the fluorescence wavelength of the nanoprobe is 1170 nm in the near infrared-II region. Its size is 139.6 nm, it has good dispersibility, and it has low cellular toxicity at concentrations up to 25 µg mL(-1) Ag. In vivo experiments revealed that the probe has a rather long circulation time (blood half-life of 5.7 hours), and the tissue histopathological tests show that it is not obviously harmful to major organs' normal function. Biochemical analysis (glutamic pyruvic transaminase and glutamic oxaloacetic transaminase levels) and blood analysis (white blood cell, red blood cell, hemoglobin and blood platelet counts) reveal that it has little influence on blood within 15 days of administration. When injected into HeLa xenograft nude mice by the tail vein, the probe elicited intensely enhanced fluorescence and X-ray computed tomography (CT) signals in the tumors after 24 hours, and the structure, size and position of tumor tissue were shown clearly. In a word, the probe has good tumor targeting capabilities, and it has significant value in fluorescence-CT dual-mode imaging in vivo.

Hybrid nanoprobes of bismuth sulfide nanoparticles and CdSe/ZnS quantum dots for mouse computed tomography/fluorescence dual mode imaging.

BACKGROUND: X-ray computed tomography (CT) imaging can be used to reveal the three-dimensional structure of deep tissue with high spatial resolution. However, it cannot reveal molecular or cellular changes, and has great limitations in terms of specificity and sensitivity. Fluorescence imaging technology is one of the main methods used for the study of molecular events in vivo and has important applications in life science research. Therefore, the combination of CT and fluorescence imaging is an ideal dual-modal molecular imaging method, which can provide data on both molecular function and tissue structure, and has important research value. In a previous study, Bi2S3 nanoparticles were wrapped with quantum dots in SiO2 to generate CT and fluorescence imaging. However, this type of probe led to low survival and caused innegligible in vivo toxicity in mice. Therefore, it is necessary to develop new multifunctional probes that demonstrate biocompatibility and safety in vivo. METHODS: A polyethylene glycol-phospholipid bilayer structure was used to synthesize hybrid clusters containing hydrophobic Bi2S3 nanoparticles and quantum dots for combined CT/fluorescence imaging. Mean particle size, polydispersity index, and zeta potential were used to study the stability over an 8-week test period. In vivo CT and fluorescence imaging experiments were performed, and the in vivo safety of the probe was evaluated, using histopathological, biochemical, and blood analyses. RESULTS: The probe distinctly enhanced the CT contrast and had fluorescence imaging capability. In addition, the nanocomposite hybrid clusters showed a longer circulation time (>4 h) than iobitridol. The results also showed that the Bi2S3-QD@DSPE probe had good biocompatibility and safety, and did not affect normal organ functioning. CONCLUSIONS: Bi2S3-QD@DSPE hybrid clusters exhibited remarkable performance in CT angiography and fluorescence imaging in vivo.

Multifunctional magnetic-hollow gold nanospheres for bimodal cancer cell imaging and photothermal therapy.

Multifunctional nanocomposites combining imaging and therapeutic functions have great potential for cancer diagnosis and therapy. In this work, we developed a novel theranostic agent based on hollow gold nanospheres (HGNs) and superparamagnetic iron oxide nanoparticles (SPIO). Taking advantage of the excellent magnetic properties of SPIO and strong near-infrared (NIR) absorption property of HGNs, such nanocomposites were applied to targeted magnetic resonance imaging (MRI) and photoacoustic imaging (PAI) of cancer cells. In vitro results demonstrated they displayed significant contrast enhancement for T2-weighted MRI and strong PAI signal enhancement. Simultaneously, the nanocomposites exhibited a high photothermal effect under the irradiation of the near-infrared laser and can be used as efficient photothermal therapy (PTT) agents for selective killing of cancer cells. All these results indicated that such nanocomposites combined with MRI-PAI and PTT functionality can have great potential for effective cancer diagnosis and therapy.