Associations between dietary patterns and renal impairment in individuals with diabetes: a cross-sectional study.

BACKGROUND: A variety of chronic diseases are affected by diet. To our knowledge, few studies have investigated the relationship between dietary patterns and renal impairment in individuals with diabetes within an Asian population. This study aimed to assess the relationship between renal impairment and dietary patterns in individuals with diabetes within a Chinese population. METHODS: In this cross-sectional survey, we analysed data on 1522 participants with diabetes aged 18 years or older who took part in the China National Diabetic Chronic Complications Study. We utilised the Chinese Diabetes Complications Questionnaire, including the semiquantitative food frequency questionnaire (SQFFQ). We identified three dietary patterns using factor analysis: Chinese traditional, healthy and plant-based dietary patterns, and these dietary patterns were used to classify participants into four groups based on the quartiles of their scores. A decrease in the estimated glomerular filtration rate (eGFR; <60 mL/min/1.73 m2 ) and an increase in the albumin-to-creatinine ratio (ACR; ≥3 mg/mmol) were used as indicators of renal impairment. Binary logistic regression models were used to estimate the odds ratio (OR) of the highest quartile (Q4: high intake levels of each dietary pattern) for renal impairment compared to the lowest quartile (Q1: low intake levels of each dietary pattern). RESULTS: Among the 1522 participants, there was a 5.5% prevalence of low eGFR, with prevalence rates of 5.2% in men and 5.9% in women, yet the prevalence of albuminuria was as high as 47.9%. After adjusting for confounders, participants in Q4 of the plant-based dietary pattern had a smaller OR for renal impairment than those in Q1. CONCLUSIONS: Our findings demonstrated that a plant-based dietary pattern is associated with a reduced risk of renal impairment in a population with diabetes.

Cellular and Intravital Imaging of NAD(P)H by a Red-Emitting Quinolinium-Based Fluorescent Probe that Features a Shift of Its Product from Mitochondria to the Nucleus.

NAD(P)H is a vital hydrogen donor and electron carrier involved in numerous biological processes. The development of small-molecule tools for intravital imaging of NAD(P)H is significant for further exploring their pathophysiological roles. Herein, we rationally designed a fluorescent probe NADH-R by a simple graft of pyridiniumylbutenenitrile on a 1-methylquinolinium moiety in the 3-position. Benefited from the reduction of quinolinium by NAD(P)H, this probe releases the free push-pull fluorophore NADH-RH, allowing a turn-on red-emitting fluorescence response together with an ultralow detection limit of 12 nM. Under the assistance of the probe, we first monitored exogenous and endogenous generation of NAD(P)H in living cells, subsequently observed dynamic changes of NAD(P)H levels in living cells under different metabolic perturbations, and finally visualized the declined NAD(P)H levels in live mouse brain in a stroke model. Unexpectedly, the time-dependent colocalization experiment revealed that the probe reacts with mitochondrial NAD(P)H, followed by a shift of its reduced product NADH-RH from mitochondria to the nucleus, highlighting that NADH-RH is a novel nucleus-directed dye scaffold, which would facilitate the development of nucleus-targeting fluorescent probes and drugs.

The Effect of Diabetes Management Shared Care Clinic on Glycated Hemoglobin A1c Compliance and Self-Management Abilities in Patients with Type 2 Diabetes Mellitus.

Objective: We aim to evaluate the impact of diabetes management shared care clinic (DMSCC) on glycated hemoglobin A1c (HbA1c) compliance and self-management abilities in patients with type 2 diabetes mellitus (T2DM). Methods: This study was a prospective cohort study of patients with T2DM participating in the DMSCC. At baseline and after management, the HbA1c levels were measured, the HbA1c compliance rate were calculated, and the Summary of Diabetes Self-Care Activities-6 (SDSCA-6), Diabetes Empowerment Scale-DAWN Short Form (DES-DSF), and Problem Areas in Diabetes Scale-Five-item Short Form (PAID-5) were completed. These pre- and post-management data were compared. Results: A total of 124 eligible patients were enrolled. After the diabetes management of DMSCC, the average HbA1c decreased and the HbA1c compliance rate increased significantly (P < 0.01). SDSCA-6 showed significant improvement in physical activity, glycemic monitoring, smoking (P < 0.01), and taking medication (P < 0.05). DES-DSF suggested a greater willingness to try to effectively treat diabetes (P < 0.05). PAID-5 indicated significant improvement in diabetes-related emotional distress. Conclusion: DMSCC can help patients with T2DM reduce HbA1c, increase HbA1c compliance, improve diabetes self-management behaviors, empowerment, and diabetes-related emotional distress and serve as an effective exploration and practice of diabetes self-management education and support.

Fast Imaging of Mitochondrial Thioredoxin Reductase Using a Styrylpyridinium-Based Two-Photon Ratiometric Fluorescent Probe.

Thioredoxin reductase (TrxR) is a pivotal antioxidant enzyme, but there remains a challenge for its fast imaging. This work describes the combination of a hydroxyl styrylpyridinium scaffold as the push-pull fluorophore with a carbonate-bridged 1,2-dithiolane unit as the reaction site to develop a fast mitochondrial TrxR2 probe, DSMP. It manifested a plethora of excellent properties including a rapid specific response (12 min), large Stokes shift (170 nm), ratiometric two-photon imaging, favorable binding with TrxR (Km = 12.5 ± 0.2 µM), and the ability to cross the blood-brain barrier. With the aid of DSMP, we visualized the increased mitochondrial TrxR2 activity in cancer cells compared to normal cells. This offers the direct imaging evidence of the connection between the increased TrxR2 activity and the development of cancer. Additionally, the probe allowed the visualization of the loss in TrxR2 activity in a cellular Parkinson's disease model and, more importantly, in mouse brain tissues of a middle cerebral artery occlusion model for ischemic stroke.

TNF-α/anti-TNF-α drugs and its effect on pregnancy outcomes.

Pregnancy is a complex biological process. The establishment and maintenance of foetal-maternal interface are pivotal events. Decidual immune cells and inflammatory cytokines play indispensable roles in the foetal-maternal interface. The disfunction of decidual immune cells leads to adverse pregnancy outcome. Tumour necrosis factor (TNF)-α, a common inflammatory cytokine, has critical roles in different stages of normal pregnancy process. However, the relationship between the disorder of TNF-α and adverse pregnancy outcomes, including preeclampsia (PE), intrauterine growth restriction (IUGR), spontaneous abortion (SA), preterm birth and so on, is still indefinite. In this review, we thoroughly reviewed the effect of TNF-α disorder on pathological conditions. Moreover, we summarized the reports about the adverse pregnancy outcomes (PE, IUGR, SA and preterm birth) of using anti-TNF-α drugs (infliximab, etanercept and adalimumab, certolizumab and golimumab) currently in the clinical studies. Overall, IUGR, SA and preterm birth are the most common adverse pregnancy outcomes of anti-TNF-α drugs. Our review may provide insight for the immunological treatment of pregnancy-related complication, and help practitioners make informed decisions based on the current evidences.

The relationship between serum uric acid levels and glomerular ischemic lesions in patients with Immunoglobin A nephropathy-a analytical cross-sectional study.

BACKGROUND: To investigate the relationship between serum uric acid levels and glomerular ischemic lesions in patients with immunoglobulin A nephropathy (IgAN) and the relevant risk factors. METHODS: A total of 86 patients with IgAN and normal renal functions were divided into a hyperuricemia group and a normal serum uric acid group (control group). These patients were further divided into a glomerular ischemic lesions group and a non-glomerular ischemic lesions group (control group) based on the renal biopsy results. The relationship between serum uric acid levels and glomerular ischemic lesions was analysed. RESULTS: In patients with IgAN, the prevalence or occurrence of glomerular ischemic lesions was significantly higher in the hyperuricemia group compared with the normal serum uric acid group. Elevated serum uric acid levels are independently associated with glomerular ischemic disease. CONCLUSION: Hyperuricemia in patients with IgAN may lead to glomerular ischemic lesions, and lowering serum uric acid levels may delay the progression of IgAN.

Gradient SERS Substrates with Multiple Resonances for Analyte Screening: Fabrication and SERS Applications.

Surface-enhanced Raman spectroscopy (SERS) provides a strong enhancement to an inherently weak Raman signal, which strongly depends on the material, design, and fabrication of the substrate. Here, we present a facile method of fabricating a non-uniform SERS substrate based on an annealed thin gold (Au) film that offers multiple resonances and gap sizes within the same sample. It is not only chemically stable, but also shows reproducible trends in terms of geometry and plasmonic response. Scanning electron microscopy (SEM) reveals particle-like and island-like morphology with different gap sizes at different lateral positions of the substrate. Extinction spectra show that the plasmonic resonance of the nanoparticles/metal islands can be continuously tuned across the substrate. We observed that for the analytes 1,2-bis(4-pyridyl) ethylene (BPE) and methylene blue (MB), the maximum SERS enhancement is achieved at different lateral positions, and the shape of the extinction spectra allows for the correlation of SERS enhancement with surface morphology. Such non-uniform SERS substrates with multiple nanoparticle sizes, shapes, and interparticle distances can be used for fast screening of analytes due to the lateral variation of the resonances within the same sample.

Developing Push-Pull Hydroxylphenylpolyenylpyridinium Chromophores as Ratiometric Two-Photon Fluorescent Probes for Cellular and Intravital Imaging of Mitochondrial NQO1.

This work highlights the use of push-pull hydroxylphenylpolyenylpyridinium fluorophores coupled with trimethyl lock quinone to engineer the ratiometric two-photon probes for cellular and intravital imaging of mitochondrial NAD(P)H:quinone oxidoreductase 1 (NQO1), a critical antioxidant enzyme responsible for detoxifying quinones. As a typical representative, QBMP showed favorable binding with NQO1 with a Michaelis constant of 12.74 µM and exhibited a suite of superior properties, including rapid response (4 min), large Stokes shift (162 nm), ultralow detection limit (0.9 nM), favorable two-photon cross section for the released fluorophore (70.5 GM), and deep tissue penetration (225 µm) in fixed brain tissues. More importantly, this probe was successfully applied for distinguishing different NQO1-expressing cancer and normal cells, revealing decreased NQO1 activity in a cellular Parkinson's disease model, screening NQO1 inducers as neuroprotective agents, and imaging of NQO1 in live mouse brain.

Expansion of targetable sites for the ribonucleoprotein-based CRISPR/Cas9 system in the silkworm Bombyx mori.

BACKGROUND: With the emergence of CRISPR/Cas9 technology, multiple gene editing procedures became available for the silkworm. Although binary transgene-based methods have been widely used to generate mutants, delivery of the CRISPR/Cas9 system via DNA-free ribonucleoproteins offers several advantages. However, the T7 promoter that is widely used in the ribonucleoprotein-based method for production of sgRNAs in vitro requires a 5' GG motif for efficient initiation. The resulting transcripts bear a 5' GG motif, which significantly constrains the number of targetable sites in the silkworm genome. RESULTS: In this study, we used the T7 promoter to add two supernumerary G residues to the 5' end of conventional (perfectly matched) 20-nucleotide sgRNA targeting sequences. We then asked if sgRNAs with this structure can generate mutations even if the genomic target does not contain corresponding GG residues. As expected, 5' GG mismatches depress the mutagenic activity of sgRNAs, and a single 5' G mismatch has a relatively minor effect. However, tests involving six sgRNAs targeting two genes show that the mismatches do not eliminate mutagenesis in vivo, and the efficiencies remain at useable levels. One sgRNA with a 5' GG mismatch at its target performed mutagenesis more efficiently than a conventional sgRNA with 5' matched GG residues at a second target within the same gene. Mutations generated by sgRNAs with 5' GG mismatches are also heritable. We successfully obtained null mutants with detectable phenotypes from sib-mated mosaics after one generation. CONCLUSIONS: In summary, our method improves the utility and flexibility of the ribonucleoprotein-based CRISPR/Cas9 system in silkworm.

High-transmission narrowband ultraviolet filter based on an aluminum laminated nanostructure on glass.

We present an aluminum (Al) laminated nanostructure stacked on a glass substrate to produce highly transmitted narrowband ultraviolet (UV) filters. The laminated nanostructure was mainly composed of an Al nanohole array, and each Al nanohole had a coaxial Al nanoring at the bottom. This UV filter showed a single dominant peak with a high transmission over 50% and a narrow bandwidth less than 80 nm in the 200-400 nm waveband that was achieved based on the synergy of surface plasmon resonance (SPR) and localized surface plasmon resonance (LSPR). The electric field profiles of the laminated nanostructure indicate that SPR selects the transmission wavelength and LSPR contributes to single peak. This narrowband UV filter can be utilized in UV detectors.

Hsa_circ_0000301 facilitates the progression of cervical cancer by targeting miR-1228-3p/IRF4 Axis.

BACKGROUND: With the broadened application of gene expression profiles analysis, the role of miRNA and circRNA are of increasing concern in recent years, especially during the pathogenesis of cancer. However, to date, the reported on this area in cervical cancer are limited. METHOD: In this study, Weighted Gene Co-Expression Network Analysis (WGCNA) and differential gene expression analysis were utilized to screen out differentially expressed (DE) circular RNAs in cervical cancer, and then we predicted and screened the combined microRNAs (miRNA) and downstream mRNAs to construct circular (circ)RNA-miRNA-mRNA network. RESULT: As a result, a regulatory circular (circ)RNA-miRNA-mRNA with 1 circRNA node, 4 miRNA nodes, 135 mRNA nodes were constructed in an attempt to provide novel biomarkers for the pathogenesis of cervical cancer. In addition, enrichment analysis including Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were performed on mRNAs in the network. After further screening of mRNAs by two online databases of GEPIA2 and RNAyhrid, precise target genes were obtained. Next, we screened out four target genes (CXCL16, IRF4, OAS3 and PTGER3) by constructing the protein-protein interaction (PPI) network, and mapped them to the initial network to reconstruct the circRNA-miRNA-mRNA network. Notably, the low expression of IRF4 was demonstrated to be associated with a significantly poorer overall survival in the GEPIA2 database, which was also verified by the immunofluorescence of the sections in Human Protein Atlas (HPA). The upstream miRNA corresponding to IRF4 is hsa-miR-1228-3p. CONCLUSION: From above concern, it can conclude that hsa_circ_0000301/hsa-miR-1228-3p/IRF4 may be involved in the occurrence and development of cervical cancer. However, the specific mechanism should be further studied and confirmed.

Pseudoxanthomonas beigongshangi sp. nov., a novel bacteria with predicted nitrite and nitrate reduce ability isolated from pit mud of Baijiu.

A Gram-negative and rod-shape bacterium designated REN9T, was isolated from pit mud of Baijiu in Sichuan, China. The 16S rRNA sequence of strain REN9T had a high similarity to Pseudoxanthomonas indica P15T (99.21%), P. mexicana AMX 26BT (97.74%) and P. japonensis 12-3T (97.43%). Phylogenetic analysis showed that REN9T belongs to the genus Pseudoxanthomonas and formed distinct cluster. The ANI and DDH values between strains REN9T and P15T were 80.94% and 24%, respectively. Strain REN9T grew optimally at 37 °C, pH 7.0 and 2% NaCl. PE (phosphatidylethanolamine), PG (phosphatidylglycerol) and DPG (diphosphatidylglycerol) were the major polar lipids of REN9T. Ubiquinone 8 (Q-8) was the predominant quinone, and Iso-C15:0 (64.32%), anteiso-C15:0 (12.04%) and Iso-C14:0 (4.56%) were the majority fatty acids. The DNA G + C content of strain REN9T was 67.3 mol%. Genomic analysis showed that strain REN9T had two secondary metabolite biosynthesis gene clusters. Moreover, strain REN9T had 43 glycoside hydrolases, 41 glycosyl transferases and 41 carbohydrate esterases. Based on the polyphasic taxonomic analysis, strain REN9T is recommended as a novel species within the genus Pseudoxanthomonas, for which the name Pseudoxanthomonas beigongshangi is proposed. The type stain is REN9T (= JCM 33961T = GDMCC 1.2210T).

Umezawaea beigongshangensis sp. nov., Isolated From the Mash of Baijiu.

A Gram-positive, aerobic and non-motile actinobacterial strain, designated REN6T, was isolated from the mash of Baijiu (Chinese spirits, a kind of distilling liquor is made by cooking, saccharification, fermentation and distillation) collected from Sichuan Province Region, China, and characterized using a polyphasic approach. Morphological and chemotaxonomic properties of strain REN6T were consistent with the description of the genus Umezawaea, such as the spore arrangement, the abundant aerial mycelium and the fragmented substrate mycelium. The diamino acid of peptidoglycan is meso-diaminopimelic acid. The diagnostic phospholipids were DPG (diphosphatidylglycerol), PG (phosphatidylglycerol), PI (phosphatidylinositol), PE (phosphatidylethanolamine). The major fatty acids were C16:0, C17:0, Iso-C16:0, Iso-C16:1 H, C17:1ω6c and C17:1 ω8c. Menaquinone-9 (MK-9) (H4) was the predominant menaquinones. The genomic DNA G + C contents were 72.7 mol%. Phylogenetic analysis based on 16S rDNA gene sequences demonstrated that strain REN6T should also be classified in the genus Umezawaea, with U. tangerina (98.7%) and U. endophytica (98.7%). However, it can be distinguished from the closest strains U. tangerina JCM 10302T based on the low levels of DNA-DNA hybridization 22.1%. Based upon the morphological, physiological, chemotaxonomic and molecular characteristics differences from other members of the genus, a novel species, Umezawaea beigongshangensis sp. nov., is proposed, with REN6T (= JCM 33954T = CGMCC 19205T) as the type strain.

The treatment of macrolide-resistant Mycoplasma pneumoniae pneumonia in children.

WHAT IS KNOWN AND OBJECTIVE: In recent years, the resistance of Mycoplasma pneumoniae to macrolide antibiotics has increased significantly. The health systems are facing significant challenges in carrying out the diagnosis and treatment of refractory Mycoplasma pneumoniae pneumonia in children. Levofloxacin is suitable for treating infectious diseases in various systems but limited in children due to arthropathy issues in weight-bearing joints. This study aimed to evaluate the efficacy and safety of levofloxacin in children with macrolide-resistant Mycoplasma pneumoniae pneumonia. METHODS: We retrospectively enrolled six confirmed cases of refractory Mycoplasma pneumoniae pneumonia who were admitted in the paediatric respiratory ward of Shandong provincial hospital Affiliated to Shandong first Medical University between 1st January 2020 and 29th February 2020. Levofloxacin was given to the patients through the intravenous or oral route as per the following dosages :<5 years, 8-10 mg/kg q12 h; >5 years, 8-10 mg/kg, qd for 10 days. The clinical data were collected and analysed. RESULTS AND DISCUSSION: The average age of the enrolled cases was six years and nine months (range, four years, and seven months to eleven years and seven months). All cases were found to be drug-resistant and were treated with azithromycin combined with antibacterial drugs. Levofloxacin was used in the patient's refractory to macrolide antibiotics. The temperature of five cases returned to normal 1-2 days after treatment with levofloxacin, and the imaging of the four cases showed expected improvements. The gastrointestinal symptoms, neurological manifestations, joint symptoms, blood parameters, liver and kidney functions, and exercise conditions of the children were closely monitored. The follow-up time of the patients ranged from one week to five months. No drug-related adverse reactions were observed in patients during treatment or during follow-up. WHAT IS NEW AND CONCLUSION: The clinical symptoms and imaging significantly improved after treatment with levofloxacin, and no drug-related adverse reactions were observed. Levofloxacin proved to be an effective and safe drug in the treatment of children with macrolide-resistant mycoplasma pneumonia. This study will provide a reference for evaluating the efficacy and safety of levofloxacin in the paediatric population.

[Impacts of the expressions of Fas and FasL mRNA and proteins on sperm concentration and motility].

Objective: To investigate the influence of the expressions of apoptosis-related Fas and FasL mRNA and proteins on sperm concentration and motility. METHODS: We collected semen samples from 80 adult males and divided them into four groups of an equal number according to sperm concentration and the percentage of progressively motile sperm (PMS): normal, asthenospermia (AS), oligozoospermia (OS) and oligoasthenospermia (OAS). We examined the routine semen parameters, the levels of Fas and FasL proteins and the expressions of Fas and FasL genes in different groups. RESULTS: The sperm concentrations in the normal, AS, OS and OAS groups were (68.11 ± 35.49), (92.21 ± 60.96), (8.55 ± 2.82) and (5.96 ± 3.80) ×106/ml, respectively, and the percentages of PMS were (49.40 ± 13.86)%, (22.12 ± 7.13)%, (40.77 ± 8.41)% and (14.53 ± 9.74), respectively. The Fas protein level was significantly higher in the AS, OS and OAS than in the normal group (ï¼»425.03 ± 50.56ï¼½, ï¼»442.32 ± 84.88ï¼½ and ï¼»448.42 ± 84.79ï¼½ vs ï¼»381.07 ± 52.37ï¼½ pg/ml, P < 0.05), correlated negatively with sperm concentration (r = －0.377, P < 0.01) and PMS (r = －0.350, P < 0.01), but exhibited no statistically significant differences between the former three and latter group (ï¼»166.98 ± 27.39ï¼½, ï¼»169.51 ± 32.62ï¼½ and ï¼»171.46 ± 32.61ï¼½ vs ï¼»167.49 ± 29.91ï¼½ pg/ml, P > 0.05). The relative levels of the Fas gene in the normal, AS, OS and OAS groups were 1, (0.88 ± 1.17), (2.55 ± 2.11) and (0.69 ± 0.90) respectively, lower in the AS and OAS than in the normal group, and positively correlated with sperm motility; those of the FasL gene were 1, (1.99 ± 1.81), (2.08 ± 2.06) and (2.03 ± 2.23) respectively, higher in the OS and OAS than in the normal group, and negatively correlated with sperm motility. Compared with the normal group, the expressions of Fas and FasL were down-regulated in the AS but up-regulated in the OS group; the expression of Fas, however, was down-regulated and that of FasL up-regulated in the OAS group. The expression of Fas mRNA was positively correlated with the percentage of PMS (r = 0.355, P = 0.01) and total sperm motility (r = 0.358, P < 0.01), while sperm concentration negatively correlated with the expression FasL mRNA (r = －0.305, P < 0.05). There was no significant correlation between the other parameters. CONCLUSIONS: Fas and FasL are differentially expressed in normal, asthenospermia, oligozoospermia and oligoasthenospermia males. Their up-regulated expressions may promote the apoptosis of spermatogenic and sperm cells and induce oligozoospermia, while their down-regulated expressions may indicate the failure of abnormal spermatogenic and sperm cells to immediately undergo programmed death, which can be one of the causes of asthenospermia.

Identification, expression, and artificial selection of silkworm epigenetic modification enzymes.

BACKGROUND: Understanding the genetic basis of phenotype variations during domestication and breeding is of great interest. Epigenetics and epigenetic modification enzymes (EMEs) may play a role in phenotypic variations; however, no comprehensive study has been performed to date. Domesticated silkworm (Bombyx mori) may be utilized as a model in determining how EMEs influence domestication traits. RESULTS: We identified 44 EMEs in the genome of silkworm (Bombyx mori) using homology searching. Phylogenetic analysis showed that genes in a subfamily among different animals were well clustered, and the expression pattern of EMEs is constant among Bombyx mori, Drosophila melanogaster, and Mus musculus. These are most highly expressed in brain, early embryo, and internal genitalia. By gene-related selective sweeping, we identified five BmEMEs under artificial selection during the domestication and breeding of silkworm. Among these selected genes, BmSuv4-20 and BmDNMT2 harbor selective mutations in their upstream regions that alter transcription factor-binding sites. Furthermore, these two genes are expressed higher in the testis and ovary of domesticated silkworm compared to wild silkworms, and correlations between their expression pattern and meiosis of the sperm and ova were observed. CONCLUSIONS: The domestication of silkworm has induced artificial selection on epigenetic modification markers that may have led to phenotypic changes during domestication. We present a novel perspective to understand the genetic basis underlying animal domestication and breeding.

Cooperation of ESIPT and ICT Processes in the Designed 2-(2'-Hydroxyphenyl)benzothiazole Derivative: A Near-Infrared Two-Photon Fluorescent Probe with a Large Stokes Shift for the Detection of Cysteine and Its Application in Biological Environments.

A rationally designed near-infrared two-photon fluorescent probe (SDP-A) for selectively detecting cysteine (Cys) has been developed based on a newly designed conjugation-enhanced 2-(2'-hydroxyphenyl)benzothiazole derivative as the fluorophore, an acrylate moiety as the Cys reaction site, and an N-methylpyridinium scaffold as both the unit of organelle targeting and improving water solubility. The probe SDP-A alone essentially emitted no fluorescence, whereas it achieved a superb near-infrared fluorescence emission (713 nm) enhancement within 15 min with a significant Stokes shift (302 nm) in the presence of Cys. The photoluminescence mechanism of the probe SDP-A toward Cys was modulated by excited-state intramolecular proton transfer (ESIPT) and intramolecular charge transfer (ICT) processes. It exhibited high selectivity and sensitivity (LOD = 102 nM) for monitoring Cys over other analytes such as Hcy/GSH/H2S owing to a specific conjugate addition-cyclization reaction between Cys and the acrylate moiety. More importantly, the released fluorophore SDP exhibits elevated quantum yields (1.52-18.17%) in different solvents and strong two-photon excited fluorescence with a sizeable two-photon action cross-section (Φ) of 213.5 GM at 820 nm in acetonitrile-PBS medium, which is highly desirable for two-photon fluorescence imaging of the living samples. Therefore, SDP-A was successfully applied to the imaging of Cys in live cells, zebrafish, mouse brain, and abdominal cavity down to a depth of more than 200 µm using a one/two-photon fluorescence microscope.

Intestinal morphology, immunity and microbiota response to dietary fibers in largemouth bass, Micropterus salmoide.

This study is aimed at identifying the effects of dietary fiber on gut health, as well as the association between that understanding and fiber consumption in fish. A total of 300 juvenile largemouth bass (micropterus salmoides, initial average weight: 15.38 ± 0.16g) were randomly divided into three treatment groups (4 replicates per group). Fish were fed with isoproteic and isolipidic diets containing 0% (low fiber, LF), 4% (moderate fiber, MF) and 8% (high fiber, HF) soybean fiber, respectively. The intestine and intestinal content of test fish per treatment group after 56 days of treatment were sampled. The results showed that the anterior intestinal sections had normal histological architecture, and no considerable damage or inflammation was observed in any histological section from all subjects examined. Curiously, fish fed the MF diet had better histological alterations than the other treatments. Meanwhile, the intestinal antioxidant capacity in the MF group was significantly promoted when compared to the other groups, as well as up-regulated expression of antioxidant-related genes including sod, cat and gpx with increasing dietary fiber concentrations. Importantly, the administrations of MF diet remarkably elevated largemouth bass innate immune parameters include intestinal inducible nitric oxide synthase (iNOS) activity, nitric oxide (NO) and total protein content. Similarly, dietary administrations of fiber down-regulated notablely the expression of pro-inflammatory cytokines including IL-8, IL-1ß and TNFα, whereas up-regulated tolerogenic cytokine IL-10 and TGF-ß1 mRNA levels. In addition, dietary fibers also modulated the community structure of the intestinal microbiota by significantly altering bacterial diversity. Dietary supplemental fibers regulated intestinal microbiota in largemouth bass, characterized by a reduced abundance of Fusobacteria along with increased abundances of Proteobacteria and Firmicutes. Taken together, the present results suggested that moderate fiber supplementation was beneficial to promoting intestinal health status of fish through antioxidant and anti-inflammatory effects, which could be at least partially responsible by the modulation of gut microbial composition.

[Regulatory effect of Qianliexiao Decoction on TGF-ß1, Smad4 and Smad 7 signaling pathways and the proliferation and apoptosis of prostate fibroblasts in mice with experimental autoimmune prostatitis].

OBJECTIVE: To study the effect of intervening in the signal transduction pathways of TGF-ß1, Smad4 and Smad7 with Qianliexiao Decoction (QLX) on the proliferation and apoptosis of prostate fibroblasts (PrF) in mice with experimental autoimmune prostatitis (EAP). METHODS: A model of EAP with damp-heat syndrome was established in C57BL/6 mice by immunization induction combined with the TCM modeling method. The prostate tissue of the mice was harvested for isolation, culturing and purification of PrFs and detection of their purity. After modeling by stimulation with a medium containing >90%-purity or 5 ng/ml TGF-ß1, the PrFs in the logarithmic growth phase were obtained and randomly divided into a blank control (serum-free medium), a model control, a positive control (medium containing 5 ng/ml TGF-ß1), a low-dose QLX (serum containing 5% QLX), a medium-dose QLX (serum containing 10% QLX), and a high-dose QLX group (serum containing 20% QLX). After 24 hours of intervention, the proliferation of the PrFs was measured, the protein expressions of TGF-ß1, Smad4, Smad7, p-Smad4 and p-Smad7 detected by Western blot, their mRNA expressions determined by qPCR, and the apoptosis of the PrFs examined by flow cytometry. RESULTS: After induction with TGF-ß1, the proliferation of the PrFs was significantly increased in the positive control (P < 0.05), but inhibited in the medium- and low-dose QLX groups (P < 0.05) and even more significantly in the high-dose QLX group as compared with that in the model control (P < 0.01). The expressions of Smad4, p-Samd7 and TGF-ß1 proteins in the PrFs were remarkably higher in the positive control than in the model control group (P < 0.05), while those of p-Smad4 and TGF-ß1 markedly lower (P < 0.01) and that of p-Smad7 dramatically higher in the QLX intervention groups than in the positive control (P < 0.01), in an evident dose-dependent manner. In comparison with the model control group, the high-dose QLX group exhibited a significant decrease in the mRNA expression of Smad4 (P < 0.05) but all the three QLX groups showed a dramatic increase in those of Smad7 (P < 0.05) and TGF-ß1 (P < 0.01). The mRNA expression of Smad4 was markedly down-regulated in the high-dose QLX group compared with that in the positive control (P < 0.05), that of Smad7 up-regulated in the model control and QLX groups (P < 0.01), and that of TGF-ß1 down-regulated in the three QLX groups (P < 0.01). The apoptosis rate of the PrFs was significantly higher in the QLX groups than in the model control (P < 0.05) in a dose-dependent manner, but showed no statistically significant difference between the model control and the positive control groups (P > 0.05). CONCLUSIONS: TGF-ß1 can stimulate the proliferation of PrFs, up-regulate the expressions of TGF-ß1 and p-Smad4, and down-regulate that of p-Smad7, while QLX can inhibit the proliferation of PrFs in a dose-dependent manner by decreasing the expressions of TGF-ß1 and p-Smad4, increasing that of p-Smad7, and thereby suppressing TGF-ß1-induced proliferation of PrFs.

Up-regulation of the active form of small GTPase Rab13 promotes macroautophagy in vascular endothelial cells.

The importance of macroautophagy (hereafter referred to as autophagy) in vascular endothelial cell (VEC) biology and dysfunction is increasingly recognized, but the molecular mechanisms of autophagy in VECs in the presence of serum are still poorly understood. Previously, we identified pterostilbene as a potent autophagy inducer of VECs in the presence of serum. In this study, we used pterostilbene as a tool to induce VEC autophagy and identified the differentially expressed genes using high-throughput DAN microarray. The small GTPase Ras-related protein in brain 13 (Rab13) was found to be the most significantly up-regulated gene in pterostilbene-treated human umbilical VECs (HUVECs). Knockdown of Rab13 blocked pterostilbene-induced mTOR inhibition and autophagy, whereas overexpression of the GTP-containing active form of Rab13 induced mTOR inhibition and autophagy in HUVECs. Using a combination of immunofluorescence and co-immunoprecipitation (co-IP) assays, we demonstrated that pterostilbene or up-regulation of the active form of Rab13 promoted the interaction between Rab13 and growth factor receptor-bound protein 2 (Grb2). Knockdown of Grb2 suppressed pterostilbene or up-regulation of the active form of Rab13-induced autophagy. Further mechanistic studies revealed that Rab13 activated the downstream AMP-activated protein kinase (AMPK) and blocked mammalian target of rapamycin (mTOR) signaling by its functional interaction with Grb2 to regulate autophagy in HUVECs. Our study firmly establishes Rab13 as a novel regulator of autophagy in VECs under nutrient-enriched conditions.