ABSTRACT
RNA 5-methylcytosine (m5C) is an abundant chemical modification in mammalian RNAs and plays crucial roles in regulating vital physiological and pathological processes, especially in cancer. However, the dysregulation of m5C and its underlying mechanisms in non-small cell lung cancer (NSCLC) remain unclear. Here we identified that NSUN2, a key RNA m5C methyltransferase, is highly expressed in NSCLC tumor tissue. We found elevated NSUN2 expression levels strongly correlate with tumor grade and size, predicting poor outcomes for NSCLC patients. Furthermore, RNA-seq and subsequent confirmation studies revealed the antioxidant-promoting transcription factor NRF2 is a target of NSUN2, and depleting NSUN2 decreases the expression of NRF2 and increases the sensitivity of NSCLC cells to ferroptosis activators both in vitro and in vivo. Intriguingly, the methylated-RIP-qPCR assay results indicated that NRF2 mRNA has a higher m5C level when NSUN2 is overexpressed in NSCLC cells but shows no significant changes in the NSUN2 methyltransferase-deficient group. Mechanistically, we confirmed that NSUN2 upregulates the expression of NRF2 by enhancing the stability of NRF2 mRNA through the m5C modification within its 5'UTR region recognized by the specific m5C reader protein YBX1, rather than influencing its translation. In subsequent rescue experiments, we show knocking down NRF2 diminished the proliferation, migration, and ferroptosis tolerance mediated by NSUN2 overexpression. In conclusion, our study unveils a novel regulatory mechanism in which NSUN2 sustains NRF2 expression through an m5C-YBX1-axis, suggesting that targeting NSUN2 and its regulated ferroptosis pathway might offer promising therapeutic strategies for NSCLC patients.
Subject(s)
Carcinoma, Non-Small-Cell Lung , Ferroptosis , Lung Neoplasms , NF-E2-Related Factor 2 , Animals , Humans , Mice , Carcinoma, Non-Small-Cell Lung/metabolism , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/pathology , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , Lung Neoplasms/genetics , Methyltransferases/metabolism , Methyltransferases/genetics , Mice, Nude , NF-E2-Related Factor 2/metabolism , NF-E2-Related Factor 2/genetics , Male , Female , Middle AgedABSTRACT
Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas12a nucleases have emerged as a promising alternative to CRISPR-Cas9 in gene editing and expression regulation. However, the adoption of Cas12a has been hindered due to general off-target activities and limited efficiency. Here, we utilized a hybrid engineered Cas12a variant and hairpin-spacer crRNAs (h-CAP) to enhance the specificity and efficiency of the CRISPR-Cas12a system. Leveraging the h-CAP strategy, we demonstrate both single-base-specific and multiplex gene expression regulation in human cells.
Subject(s)
CRISPR-Cas Systems , Gene Editing , Humans , CRISPR-Cas Systems/genetics , Endonucleases/metabolismABSTRACT
Pathological cardiac hypertrophy is a complex process that often leads to heart failure. Label-free proteomics has emerged as an important platform to reveal protein variations and to elucidate the mechanisms of cardiac hypertrophy. Endomyocardial biopsy is a minimally invasive technique for sampling cardiac tissue, but it yields only limited amounts of an ethically permissible specimen. After regular pathological examination, the remaining trace samples pose significant challenges for effective protein extraction and mass spectrometry analysis. Herein, we developed trace cardiac tissue proteomics based on the anchor-nanoparticles (TCPA) method. We identified an average of 6666 protein groups using â¼50 µg of myocardial interventricular septum samples by TCPA. We then applied TCPA to acquire proteomics from patients' cardiac samples both diagnosed as hypertrophic hearts and myocarditis controls and identified significant alterations in pathways such as regulation of actin cytoskeleton, oxidative phosphorylation, and cGMP-PKG signaling pathway. Moreover, we found multiple lipid metabolic pathways to be dysregulated in transthyretin cardiac amyloidosis compared to other types of cardiac hypertrophy. TCPA offers a new technique for studying pathological cardiac hypertrophy and can serve as a platform toolbox for proteomic research in other cardiac diseases.
Subject(s)
Myocardium , Nanoparticles , Proteomics , Proteomics/methods , Humans , Myocardium/metabolism , Myocardium/pathology , Myocardium/chemistry , Nanoparticles/chemistry , Cardiomegaly/metabolism , Cardiomegaly/pathology , Cardiomegaly/diagnosis , Amyloidosis/metabolism , Amyloidosis/pathology , Amyloid Neuropathies, FamilialABSTRACT
Characterising the microstructure of foams is an important task for improving foam manufacturing processes and building foam numerical models. This study proposed a method for measuring the thickness of individual cell walls of closed-cell foams in micro-CT images. It comprises a distance transform on CT images to obtain thickness information of cell walls, a watershed transform on the distance matrix to locate the midlines of cell walls, identifying the intersections of midlines of cell walls by examining how many regions each pixel on the midlines of cell walls connects with, disconnecting and numbering the midlines of cell walls, extracting the distance values of the pixels on the midlines (or midplanes) of cell walls, and calculating the thickness of individual cell walls by multiplying the extracted distance values by two. Using this method, the thickness of cell walls of a polymeric closed-cell foam was measured. It was found that cell wall thickness measured in 2D images shows larger average values (around 1.5 times) and dispersion compared to that measured in volumetric images.
Subject(s)
Polymers , X-Ray Microtomography/methodsABSTRACT
Mass spectrometry has emerged as a mainstream technique for label-free proteomics. However, proteomic coverage for trace samples is constrained by adsorption loss during repeated elution at sample pretreatment. Here, we demonstrated superparamagnetic composite nanoparticles functionalized with molecular glues (MGs) to enrich proteins in trace human biofluid. We showed high protein binding (>95 %) and recovery (≈90 %) rates by anchor-nanoparticles. We further proposed a Streamlined Workflow based on Anchor-nanoparticles for Proteomics (SWAP) method that enabled unbiased protein capture, protein digestion and pure peptides elution in one single tube. We demonstrated SWAP to quantify over 2500 protein groups with 100 HEK 293T cells. We adopted SWAP to profile proteomics with trace aqueous humor samples from cataract (n=15) and wet age-related macular degeneration (n=8) patients, and quantified ≈1400 proteins from 5â µL aqueous humor. SWAP simplifies sample preparation steps, minimizes adsorption loss and improves protein coverage for label-free proteomics with previous trace samples.
Subject(s)
Proteins , Proteomics , Humans , Proteomics/methods , Peptides , Mass Spectrometry/methods , Magnetic Iron Oxide Nanoparticles , Proteome/analysisABSTRACT
Patients with hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) are characterized by immune paralysis and susceptibility to infections. Macrophages are important mediators of immune responses can be subclassified into two main phenotypes: classically activated and alternatively activated. However, few studies have investigated changes to macrophage polarization in HBV-related liver diseases. Therefore, we investigated the functional status of monocyte-derived macrophages (MDMs) from patients with mild chronic hepatitis B (n = 226), HBV-related compensated cirrhosis (n = 36), HBV-related decompensated cirrhosis (n = 40), HBV-ACLF (n = 62) and healthy controls (n = 10), as well as Kupffer cells (KCs) from patients with HBV-ACLF (n = 3). We found that during the progression of HBV-related liver diseases, the percentage of CD163+ CD206+ macrophages increased, while the percentage of CD80+ human leukocyte antigen-DR+ macrophages decreased significantly. MDMs and KCs mainly exhibited high CD163+ CD206+ expression in patients with HBV-ACLF, which predicted poor clinical outcome and higher liver transplantation rate. Transcriptome sequencing analysis revealed that chloride intracellular channel-3 (CLIC3) was reduced in patients with HBV-ACLF, indicating a poor prognosis. To further study the effect of CLIC3 on macrophage polarization, human monocytic THP-1 cell-derived macrophages were used. We found that classical and alternative macrophage activation occurred through nuclear factor kappa B (NF-κB) and phosphoinositide 3-kinase/protein kinase B pathways, respectively. CLIC3 suppression inhibited NF-κB activation and promoted the alternative activation. In conclusion, macrophage polarization gradually changed from classically activated to alternatively activated as HBV-related liver diseases progressed. Both CLIC3 suppression and increased alternatively activated macrophage percentage were potential indicators of the poor prognosis of patients with HBV-ACLF.
Subject(s)
Acute-On-Chronic Liver Failure , Chloride Channels/metabolism , Hepatitis B, Chronic , Chlorides , Hepatitis B virus , Hepatitis B, Chronic/complications , Humans , Liver Cirrhosis , Macrophage Activation , Macrophages , NF-kappa B , Phosphatidylinositol 3-KinasesABSTRACT
Thermoelectric materials are considered promising candidates for thermal energy conversion. This study presents the fabrication of Zn- and Ce-alloyed In2O3with a porous structure. The electrical conductivity was improved by the alloying effect and an ultra-low thermal conductivity was observed owing to the porous structure, which concomitantly provide a distinct enhancement ofZT. However, SiO2nanoparticle additives react with the matrix to form a third-phase impurity, which weakens the electrical conductivity and increases the thermal conductivity. A thermoelectric module was constructed for the purpose of thermal heat energy conversion. Our experimental results proved that both an enhancement in electrical conductivity and a suppression in thermal conductivity could be achieved through nano-engineering. This approach presents a feasible route to synthesize porous thermoelectric oxides, and provides insight into the effect of additives; moreover, this approach is a cost-effective method for the fabrication of thermoelectric oxides without traditional hot-pressing and spark-plasma-sintering processes.
ABSTRACT
BACKGROUND: Neutrophilic inflammation in the airway is a hallmark of bronchiectasis. Neutrophil extracellular traps (NETs) have been reported to play an important role in the occurrence and development of bronchiectasis. Neutrophil side fluorescence is one of the characteristics of neutrophils that can reflect the activation of neutrophils and the formation of NETs. OBJECTIVE: To explore the relationship between the values of neutrophil side fluorescence (NEUT-SFL) in the peripheral blood of bronchiectasis patients, and the severity of the disease. METHODS: 82 patients with bronchiectasis from the Department of Respiratory and Critical Medicine, at the Third Affiliated Hospital of Southern Medical University and were scored with Bronchiectasis Severity Index (BSI) (2019-2021). The clinical data such as the value of NEUT-SFL, neutrophil count, C-reactive protein, and procalcitonin levels were collected and retrospectively analyzed. NEUT-SFL values neutrophil count from 28 healthy subjects were also used to ascertain cut-off values. RESULTS: Based on the BSI scores, patients were divided into three categories as mild (32%), moderate (29%), and severe (39%). Our results showed that the values of NEUT-SFL were higher in bronchiectasis patients compared to healthy controls. The levels of NEUT-SFL positively correlated with the high BSI scores in patients (P = 0.037, r = 0.23) and negatively correlated with the lung function in these patients (r = - 0.35, P = 0.001). The area under the ROC curve was 0.813, the best cut-off was 42.145, indicating that NEUT-SFL values > 42.145 can potentially predict the severity of bronchiectasis. CONCLUSIONS: The values of NEUT-SFL in the peripheral blood can be used for predicting the severity of bronchiectasis.
Subject(s)
Bronchiectasis , Neutrophils , C-Reactive Protein , Humans , Leukocyte Count , Neutrophils/physiology , Retrospective StudiesABSTRACT
Atrial fibrillation (AF) is characterized by potentiated growth of atrial fibroblasts and excessive deposition of the extracellular matrix. Atrial fibrosis has emerged as a hallmark of atrial structural remodeling linked to AF. Nonetheless, the specific mechanism underlying the progression of atrial fibrosis to AF is still largely unknown. MFGE8 (milk fat globule-EGF factor 8) is a soluble glycoprotein associated with many human diseases. Recently, a number of studies revealed that MFGE8 plays a crucial role in heart disease. Yet, MFGE8 regulation and function in the process of atrial fibrosis and vulnerability to AF remain unexplored. In this study, we found that the expression of MFGE8 was downregulated in the atriums of patients with AF compared with individuals without AF. In addition, the expression of MFGE8 was lower in atriums of angiotensin II (Ang-II)-stimulated rats as compared with the sham group. In vitro, silencing of MFGE8 by small interfering RNA significantly increased Ang-II-induced atrial fibrosis, whereas administration of recombinant human MFGE8 (rhMFGE8) attenuated the atrial fibrosis. Moreover, we found that the activated TGF-ß1/Smad2/3 pathway after Ang-II treatment was significantly potentiated by the MFGE8 knockdown but inhibited by rhMFGE8 in vitro. Inhibition of integrin ß3 which is the receptor for MFGE8, suppressed the TGF-ß1/Smad2/3 activating effects of the MFGE8 knockdown in Ang-II-treated rat atrial fibroblasts. Finally, we administered rhMFGE8 to rats; it attenuated atrial fibrosis and remodeling and further reduced AF vulnerability induced by Ang-II, indicating that MFGE8 might have the potential both as a novel biomarker and as a therapeutic target in atrial fibrosis and AF.
Subject(s)
Antigens, Surface/metabolism , Atrial Fibrillation/metabolism , Atrial Fibrillation/pathology , Heart Atria/pathology , Milk Proteins/metabolism , Signal Transduction , Smad Proteins/metabolism , Transforming Growth Factor beta1/metabolism , Angiotensin II , Animals , Antigens, Surface/genetics , Atrial Fibrillation/genetics , Cell Movement/drug effects , Cell Proliferation/drug effects , Cells, Cultured , Fibroblasts/metabolism , Fibroblasts/pathology , Fibrosis , Gene Expression Regulation/drug effects , Humans , Integrin beta3/metabolism , Male , Milk Proteins/genetics , Models, Biological , Protein Binding/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Small Interfering/metabolism , Rats, Sprague-Dawley , Recombinant Proteins/pharmacology , Signal Transduction/drug effectsABSTRACT
BACKGROUND: ß-cell dysfunction is one of the core pathogenetic mechanisms of type 2 diabetes mellitus (T2DM). However, there are currently no effective therapeutic strategies to preserve ß-cell mass and function. The role of islet macrophage phenotype reprogramming in ß-cell dysfunction has attracted great attention. Given that advanced glycation end products (AGEs) are major pathogenic factors in T2DM, we investigated the effect of AGEs on macrophage activation and their role in ß-cell dysfunction. METHODS: We examined cytokine secretion, M1 and M2 macrophage-associated marker expression and MAPK phosphorylation levels in AGEs-stimulated macrophages. MIN6 cells were cocultured with AGEs-pretreated macrophages to study the effect of AGEs-induced macrophage activation on ß-cell dysfunction. RESULTS: We found that AGEs treatment significantly enhanced macrophage secretion of proinflammatory cytokines. The expression of M1 macrophage markers, such as iNOS and the surface marker CD11c, was significantly upregulated, whereas the expression of M2 macrophage markers, such as Arg1 and CD206, was reciprocally downregulated upon AGEs stimulation. AGEs treatment predominantly activated the MAPK pathway, and the inhibition of the MAPK pathway partially attenuated the AGEs-induced polarization of macrophages. In addition, coculture with AGEs-pretreated macrophages significantly inhibited the expression of molecules involved in ß-cell function and was accompanied by the impairment of glucose-stimulated insulin secretion (GSIS) in MIN6 cells. CONCLUSION: AGEs enhance the expression of proinflammatory molecules by activating the MAPK pathway. Moreover, these data imply that AGEs induce macrophage M1 phenotype polarization but restrain M2 polarization, which might contribute to ß-cell dysfunction in the pathogenesis of T2DM.
Subject(s)
Glycation End Products, Advanced/pharmacology , Macrophage Activation/drug effects , Macrophages/metabolism , Animals , Cell Line , Coculture Techniques , Cytokines/metabolism , Diabetes Mellitus, Type 2/etiology , Diabetes Mellitus, Type 2/pathology , Insulin-Secreting Cells/metabolism , Insulin-Secreting Cells/pathology , MAP Kinase Signaling System/drug effects , Macrophages/drug effects , Macrophages/immunology , MiceABSTRACT
Humanin (HN), a mitochondrial derived peptide, plays cyto-protective role under various stress. In this study, we aimed to investigate the effects of HNGF6A, an analogue of HN, on osteoblast apoptosis and differentiation and the underlying mechanisms. Cell proliferation of murine osteoblastic cell line MC3TC-E1 was examined by CCK8 assay and Edu staining. Cell apoptosis was detected by Annexin V assay under H2O2 treatment. The differentiation of osteoblast was determined by Alizarin red S staining. We also tested the expression of osteoblast phenotype related protein by real-time PCR and Western blot. The interaction between Circ_0001843 and miR-214, miR-214 and TAFA5 was examined by luciferase report assay. Circ_0001843 was inhibited by siRNA and miR-214 was suppressed by miR-214 inhibitor to determine the effects of Circ_0001843 and miR-214 on cell proliferation, apoptosis, and differentiation. HNGF6A, an analogue of HN, exerted cyto-protection and osteogenesis-promotion in MC3T3-E1 cells. The expression of osteoblast phenotype related protein was significantly induced by HNGF6A. Additionally, HNGF6A treatment decreased Circ_0001843 and increased miR-214 levels, as well as inhibited the phosphorylation of p38 and JNK. We further found that Circ_0001843 directly bound with miR-214, which in turn inhibited the phosphorylation of p38 and JNK. Furthermore, both Circ_0001843 overexpression and miR-214 knockdown significantly decreased the cyto-protection and osteogenic promotion of HNGF6A. In summary, our data showed that HNGF6A protected osteoblasts from oxidative stress-induced apoptosis and osteoblast phenotype inhibition by targeting Circ_0001843/miR-214 pathway and the downstream kinases, p38 and JNK.
Subject(s)
Apoptosis , Intracellular Signaling Peptides and Proteins/pharmacology , MicroRNAs , Osteoblasts , Oxidative Stress , RNA, Circular/metabolism , 3T3 Cells , Animals , Cell Differentiation , Mice , MicroRNAs/metabolism , Osteoblasts/cytology , PhenotypeABSTRACT
Currently, hepatitis C antiviral therapy is entering a new era with the use of direct-acting antiviral (DAA) agents. However, the precise immunological influences of DAA therapy in patients with chronic hepatitis C (CHC) are insufficiently understood. This study aimed to investigate the effects of DAA therapy on the frequency of myeloid-derived suppressor cells (MDSCs), T lymphocytes, and natural killer (NK) cells in patients with CHC. Thirty-two treatment-naive CHC patients were treated with DAA therapy, and the frequency of immune cells was analyzed by flow cytometry at various time points during and after therapy. Sixteen healthy donors were recruited for comparison. DAA therapy decreased the frequency of MDSCs and monocytic MDSCs in patients with CHC to a normal level. DAA therapy also increased the CD8+ T and NK cell levels in patients with CHC. In addition, activation (NKp30 and NKp46) and inhibitory (NKG2A) receptors on NK cells were downregulated to yield an NK cell phenotype resembling that observed in the healthy controls. This study provides insight into the normalization of immune cell levels under DAA therapy and indicates that restoration of the immune system in patients with CHC strongly supports long-term curative hepatitis C virus eradication.
Subject(s)
Antiviral Agents/therapeutic use , Hepatitis C, Chronic/drug therapy , Killer Cells, Natural/immunology , Myeloid-Derived Suppressor Cells/immunology , Adult , Female , Flow Cytometry , Hepatitis C, Chronic/pathology , Humans , Male , Middle Aged , T-Lymphocytes/immunology , Treatment OutcomeABSTRACT
The aim of the work was to study the influence of vaspin on oxidative stress-induced apoptosis of mouse mesenchymal stem cells (MSCs). MSCs originated from bone marrow of C57BL/6 mouse were treated with vaspin and/or H2O2 in a dose-dependent manner. Cellular viability detected by CCK-8 and cell apoptosis studied by flow cytometry and TUNEL assay were observed in these cells. The protein expressions of PI3K, p-PI3K, Akt, p-Akt, T-ERK1/2, p-ERK1/2, p38, p-p38, JNK, and p-JNK were tested by Western blot. Vaspin had no significant effect on cellular viability, but significantly reduced H2O2-induced apoptosis. Western blot assay showed that pretreatment with vaspin promoted the activation of p-p38. Inhibition of p38 by SB203580 suppressed the protective effect of vaspin on oxidative stress-induced apoptosis. Vaspin inhibits oxidative stress-induced apoptosis of MSCs via the activation of MAPK/p38 signaling pathway. These findings indicate that vaspin is prone to osteoporosis protection.
Subject(s)
Adipokines/metabolism , Apoptosis , Cytoprotection , MAP Kinase Signaling System , Mesenchymal Stem Cells/enzymology , Mesenchymal Stem Cells/pathology , Oxidative Stress , Serpins/metabolism , p38 Mitogen-Activated Protein Kinases/metabolism , Adipokines/pharmacology , Animals , Apoptosis/drug effects , Cell Survival/drug effects , Cytoprotection/drug effects , Hydrogen Peroxide/toxicity , MAP Kinase Signaling System/drug effects , Mesenchymal Stem Cells/drug effects , Mice, Inbred C57BL , Oxidative Stress/drug effects , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Serpins/pharmacologyABSTRACT
Restoring antiviral immunity is a promising immunotherapeutic approach to the treatment of chronic hepatitis B virus (HBV) infection. Dendritic cells play a crucial role in triggering antiviral immunity. In this study, we identified immunodominant epitopes prevalent in CD8+ T cell responses. We characterized the hierarchy of HBV epitopes targeted by CD8+ T cells following autologous monocyte-derived dendritic cell (moDC) expansion in HBV-infected subjects with distinct disease stages: treatment-naïve (TN group, n = 168), treatment with complete virological response (TR group, n = 72), and resolved HBV infection (RS group, n = 28). T cell responses against 32 HBV epitopes were measured upon moDC expansion. Several subdominant epitopes that triggered HBV-specific CD8+ T cell responses were identified. These epitopes' responses varied in individuals with different disease stages. Moreover, the most immunodominant and immunoprevalent epitope included the envelope residues 256-270 (Env256-270), corresponding to amino acid residues 93-107 in the small HBV surface protein (SHBs) across three patient groups. The frequency of Env256-270-specific interferon-γ-producing T cells was the highest in the RS group and the lowest in the TN group. In addition, individuals with HLA-A*02:03/02:06/02:07 were capable of responding to Env256-270. Env256-270-specific CD8+ T cells tolerated amino acid variations within the epitope detected in HBV genotypes B and C. This suggests that Env256-270 in SHBs is crucial in HBV-specific T cell immunity following autologous moDC expansion. It might be a potential target epitope for dendritic-cell-based immunotherapy for CHB patients with complete viral suppression by long-term NAs treatment.
Subject(s)
Dendritic Cells/immunology , Epitopes, T-Lymphocyte/immunology , Hepatitis B Surface Antigens/immunology , Hepatitis B virus/immunology , Hepatitis B/immunology , Adult , CD8-Positive T-Lymphocytes/cytology , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/virology , Cell Differentiation , Cell Proliferation , Cells, Cultured , Dendritic Cells/chemistry , Dendritic Cells/cytology , Epitope Mapping , Epitopes, T-Lymphocyte/chemistry , Epitopes, T-Lymphocyte/genetics , Female , Hepatitis B/genetics , Hepatitis B/physiopathology , Hepatitis B/virology , Hepatitis B Surface Antigens/chemistry , Hepatitis B Surface Antigens/genetics , Hepatitis B virus/chemistry , Hepatitis B virus/genetics , Humans , Immunodominant Epitopes/chemistry , Immunodominant Epitopes/genetics , Immunodominant Epitopes/immunology , Interferon-gamma/genetics , Interferon-gamma/immunology , Male , Middle Aged , Monocytes/cytology , Monocytes/immunology , Young AdultABSTRACT
We report the latest progress in fabrication and laser performance of the fully crystalline double-clad 'Yb:YAG-core/undoped-YAG-clad' fibers grown by the hybrid crystal growth method. The single-crystalline ytterbium (Yb) doped yttrium aluminum garnet (YAG) fiber cores were grown by the laser heated pedestal growth (LHPG) method, and the single-crystalline undoped YAG claddings were grown by the liquid phase epitaxy (LPE) technique, in which the single-crystalline Yb:YAG cores were used as the growth seeds. The key parameters of the hybrid-grown 'crystalline core/crystalline clad' (C4) fibers, including material composition, crystal structure, and fiber propagation loss, were characterized. The results confirmed that the grown C4 fibers, indeed, have both the single-crystalline fiber core and single-crystalline fiber clad. By utilizing a double-clad low-loss C4 fiber as a diode-cladding-pumped laser gain medium, we realized a fiber laser with the optical-to-optical conversion efficiency of 68.7% versus the incident pump power.
ABSTRACT
In order to develop excellent germplasm resources for giant grouper (Epinephelus lanceolatus), cryopreservation of giant grouper sperm was examined in the present study. Firstly, 13 kinds of sperm dilution (ELS1-3, EM1-2, TS-2, MPRS, ELRS0-6) were prepared with physiological salt, sucrose, glucose and fetal bovine serum. The physiological parameters of ELRS3 (ratio of fast motion, ratio of slow motion, time of fast motion, time of slow motion, lifespan and motility) and ELS3 (sperm ratio of slow motion, time of slow motion and motility) were significantly higher than those of the other dilutions (P < 0.05). Secondly, after adding 15% DMSO and 10% FBS to ELRS3 and ELS3, most physiological parameters of frozen sperm were also significantly higher than the other gradients (P < 0.05), and sperm motility was as high as 63.68 ± 4.16% to74.75 ± 12.71% (fresh sperm motility, 80.70 ± 1.37% to 80.71 ± 1.49%). Mixed with the above dilutions, a final volume of 105 ml semen was cryopreserved. Finally, the sperm of giant grouper cryopreserved with cryoprotectants (ELRS3 + 15% DMSO + 10% FBS) was used for electron-microscopic observation and crossbreeding with red-spotted groupers (Epinephelus akaara). The electron-microscopic observation revealed that part of the frozen-thawed sperm was cryodamaged, e.g., flagellum fracturing and mitochondria falling out, while the ultrastructure of sperm membrane, mitochondria and flagellum remained intact. Also, the fertilization and hatchability rates of giant grouper frozen sperm and red-spotted grouper eggs were as high as 94.56% and 75.56%, respectively. Thus, a technique for cryopreservation of giant grouper sperm was successfully developed and applied to crossbreeding with red-spotted grouper eggs.
Subject(s)
Bass/physiology , Cryopreservation/veterinary , Semen Preservation/veterinary , Spermatozoa/physiology , Animals , Aquaculture , Cell Survival/drug effects , China , Crosses, Genetic , Cryoprotective Agents/adverse effects , Female , Fertilization in Vitro/veterinary , Male , Microscopy, Electron, Scanning/veterinary , Microscopy, Electron, Transmission/veterinary , Ovulation Induction/veterinary , Semen Preservation/adverse effects , Sperm Motility/drug effects , Spermatozoa/cytology , Spermatozoa/drug effects , Spermatozoa/ultrastructure , Survival AnalysisABSTRACT
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked incompletely dominant enzyme deficiency that results from G6PD gene mutations. Women heterozygous for G6PD mutations exhibit variation in the loss of enzyme activity but the cause of this phenotypic variation is unclear. We determined DNA methylation and X-inactivation patterns in 71 G6PD-deficient female heterozygotes and 68 G6PD non-deficient controls with the same missense mutations (G6PD Canton c.1376G>T or Kaiping c.1388G>A) to correlate determinants with variable phenotypes. Specific CpG methylations within the G6PD promoter were significantly higher in G6PD-deficient heterozygotes than in controls. Preferential X-inactivation of the G6PD wild-type allele was determined in heterozygotes. The incidence of preferential X-inactivation was 86.2% in the deficient heterozygote group and 31.7% in the non-deficient heterozygote group. A significant negative correlation was observed between X-inactivation ratios of the wild-type allele and G6PD/6-phosphogluconate dehydrogenase (6PGD) ratios in heterozygous G6PD Canton (r=-0.657, p<0.001) or Kaiping (r=-0.668, p<0.001). Multivariate logistic regression indicated that heterozygotes with hypermethylation of specific CpG sites in the G6PD promoter and preferential X-inactivation of the wild-type allele were at risk of enzyme deficiency.
Subject(s)
DNA Methylation , Genetic Variation , Glucosephosphate Dehydrogenase Deficiency/genetics , Glucosephosphate Dehydrogenase/genetics , Mutation, Missense , X Chromosome Inactivation , Adult , Base Sequence , CpG Islands , Female , Genotype , Glucosephosphate Dehydrogenase Deficiency/pathology , Heterozygote , Humans , Logistic Models , Molecular Sequence Annotation , Phenotype , Phosphogluconate Dehydrogenase/genetics , Promoter Regions, Genetic , Risk FactorsABSTRACT
AIM: To explore the relationship between metabolic syndrome (MS) and risk for chronic kidney disease (CKD) in a Southern Chinese population. METHODS: A cross-sectional study was conducted in 1724 community-based Southern Chinese participants from June to October 2012. The prevalence of MS (as defined by the International Diabetes Federation) and CKD (defined as an estimated glomerular filtration rate of <60 mL/min per 1.73 m(2) and/or albuminuria) was determined. The association between MS and CKD was then analyzed using STATA software. RESULTS: Metabolic syndrome was significantly associated with CKD (P < 0.001) in the unadjusted analyses as well as after adjustment for potential confounders. The unadjusted odds ratio and adjusted odds ratio for MS were 3.53 (95% confidence interval (CI) 2.62 to 4.75, P < 0.001) and 2.52 (95% CI 1.84 to 3.54, P < 0.001). When further adjusted for diabetes and hypertension, the association of MS and CKD was significant (odds ratio (OR) 1.63, 95% CI 1.15 to 2.32, P = 0.006). After adjustment for potential confounders, three components and four/five components were associated with CKD. The OR for three components and four/five components were 2.90 (95% CI 1.70 to 4.96, P < 0.001) and 3.64(95% CI 1.95 to 6.80, P < 0.001), when compared with those without components. High blood pressure, high serum triglyceride level, elevated fasting glucose level and central obesity were associated with CKD (P < 0.05). The odds ratios for elevated blood pressure, elevated serum triglyceride levels, elevated fasting glucose and central obesity were 1.80 (95% CI 1.25 to 2.62, P = 0.002), 1.56 (95% CI 1.14 to 2.14, P = 0.006), 2.54 (95% CI 1.82 to 3.57, P < 0.001), and 1.50 (95% CI 1.10 to 2.07, P = 0.01), respectively. CONCLUSION: These findings suggest that MS is associated with CKD in Southern Chinese population, which may provide important information for the overall control of these diseases.
Subject(s)
Asian People/statistics & numerical data , Metabolic Syndrome/ethnology , Renal Insufficiency, Chronic/ethnology , Adolescent , Adult , Aged , China/epidemiology , Confounding Factors, Epidemiologic , Cross-Sectional Studies , Female , Glomerular Filtration Rate , Humans , Male , Middle Aged , Odds Ratio , Prevalence , Risk Factors , Urban Population/statistics & numerical data , Young AdultABSTRACT
The burden of fungal infections on human health is increasing worldwide. Aspergillus, Candida, and Cryptococcus are the top three human pathogenic fungi that are responsible for over 90% of infection-related deaths. Moreover, effective antifungal therapeutics are lacking, primarily due to host toxicity, pathogen resistance, and immunodeficiency. In recent years, nanomaterials have proved not only to be more efficient antifungal therapeutic agents but also to overcome resistance against fungal medication. This review will examine the limitations of standard antifungal therapy as well as focus on the development of nanomaterials.
ABSTRACT
Layered double hydroxide (LDH) nanosheets as one type of two-dimensional materials have garnered increasing attention in the field of oxygen evolution reaction (OER) in recent decades. To address the challenges associated with poor conductivity and limited electron and charge transfer capability in LDH materials, we have developed a straightforward one-pot synthesis method to successfully fabricate a composite material with a microstructure resembling cabbage, which encompasses NiFe-LDH and nanocarbon (referred as NiFe-LDH@C). Atomic force microscopy (AFM) and high-resolution transmission electron microscopy (HRTEM) revealed that the monolayer NiFe-LDH with a height of ~0.5-0.8â nm is uniformly distributed and closely bonded to the carbon support, leading to a significant enhancement in conductivity and facilitating faster electron and charge transfer. Moreover, the NiFe-LDH@C exhibits a substantial number of surface defect sites, which enhances the interaction with oxygen species. This dual enhancement in charge transfer and oxygen species-mediated transfer greatly improves the catalytic OER performance, which is further corroborated by theoretical calculations. Notably, the Ni10Fe6-LDH@C with the highest concentration of surface oxygen vacancies demonstrated superior water oxidation performance, surpassing commercially available RuO2 catalysts; an OER overpotential of 231â mV@10â mA cm-2 with a Tafel slope of 71â mV dec-1 was achieved.