[Analysis of clinical features and genetic variants in three children with late-onset Multiple acyl-Coenzyme A dehydrogenase deficiency].

OBJECTIVE: To explore the clinical characteristics and genetic variants in three children with late-onset Multiple acyl-Coenzyme A dehydrogenase deficiency (MADD type Ⅲ). METHODS: Clinical data of three children diagnosed with late-onset MADD at the Children's Hospital Affiliated to Zhengzhou University between March 2020 and March 2022 were retrospectively analyzed. All children were subjected to whole exome sequencing (WES), and candidate variants were verified by Sanger sequencing. All children had received improved metabolic therapy and followed up for 1 ~ 3 years. RESULTS: The children had included 2 males and 1 female, and aged from 2 months to 11 years and 7 months. Child 1 had intermittent vomiting, child 2 had weakness in lower limbs, while child 3 had no symptom except abnormal neonatal screening. Tandem mass spectrometry of the three children showed elevation of multiple acylcarnitines with short, medium and long chains. Children 1 and 2 showed increased glutaric acid and multiple dicarboxylic acids by urine Gas chromatography-mass spectrometry (GC-MS) analysis. All children were found to harbor compound heterozygous variants of the ETFDH gene, including a paternal c.1211T>C (p.M404T) and a maternal c.488-22T>G variant in child 1, a paternal c.1717C>T (p.Q573X) and a maternal c.250G>A (p.A84T) variant in child 2, and a paternal c.1285+1G>A and maternal c.629A>G (p.S210N) variant in child 3. As for the treatment, high-dose vitamin B2, levocarnitine and coenzyme Q10 were given to improve the metabolism, in addition with a low fat, hypoproteinic and high carbohydrate diet. All children showed a stable condition with normal growth and development during the follow-up. CONCLUSION: The compound heterozygous variants of the ETFDH gene probably underlay the muscle weakness, remittent vomiting, elevated short, medium, and long chain acylcarnitine, as well as elevated glutaric acid and various dicarboxylic acids in the three children with type Ⅲ MADD.

Advancements in the impact of human microbiota and probiotics on leukemia.

The human gut microbiota is a complex ecosystem that plays a crucial role in promoting the interaction between the body and its environment. It has been increasingly recognized that the gut microbiota has diverse physiological functions. Recent studies have shown a close association between the gut microbiota and the development of certain tumors, including leukemia. Leukemia is a malignant clonal disease characterized by the uncontrolled growth of one or more types of blood cells, which is the most common cancer in children. The imbalance of gut microbiota is linked to the pathological mechanisms of leukemia. Probiotics, which are beneficial microorganisms that help maintain the balance of the host microbiome, play a role in regulating gut microbiota. Probiotics have the potential to assist in the treatment of leukemia and improve the clinical prognosis of leukemia patients. This study reviews the relationship between gut microbiota, probiotics, and the progression of leukemia based on current research. In addition, utilizing zebrafish leukemia models in future studies might reveal the specific mechanisms of their interactions, thereby providing new insights into the clinical treatment of leukemia. In conclusion, further investigation is still needed to fully understand the accurate role of microbes in leukemia.

GhHAM regulates GoPGF-dependent gland development and contributes to broad-spectrum pest resistance in cotton.

Cotton is a globally cultivated crop, producing 87% of the natural fiber used in the global textile industry. The pigment glands, unique to cotton and its relatives, serve as a defense structure against pests and pathogens. However, the molecular mechanism underlying gland formation and the specific role of pigment glands in cotton's pest defense are still not well understood. In this study, we cloned a gland-related transcription factor GhHAM and generated the GhHAM knockout mutant using CRISPR/Cas9. Phenotypic observations, transcriptome analysis, and promoter-binding experiments revealed that GhHAM binds to the promoter of GoPGF, regulating pigment gland formation in cotton's multiple organs via the GoPGF-GhJUB1 module. The knockout of GhHAM significantly reduced gossypol production and increased cotton's susceptibility to pests in the field. Feeding assays demonstrated that more than 80% of the cotton bollworm larvae preferred ghham over the wild type. Furthermore, the ghham mutants displayed shorter cell length and decreased gibberellins (GA) production in the stem. Exogenous application of GA3 restored stem cell elongation but not gland formation, thereby indicating that GhHAM controls gland morphogenesis independently of GA. Our study sheds light on the functional differentiation of HAM proteins among plant species, highlights the significant role of pigment glands in influencing pest feeding preference, and provides a theoretical basis for breeding pest-resistant cotton varieties to address the challenges posed by frequent outbreaks of pests.

Synthesis and biological evaluation of orally active anti-Trypanosoma agents.

In previous studies, we developed anti-trypanosome tubulin inhibitors with promising in vitro selectivity and activity against Human African Trypanosomiasis (HAT). However, for such agents, oral activity is crucial. This study focused on further optimizing these compounds to enhance their ligand efficiency, aiming to reduce bulkiness and hydrophobicity, which should improve solubility and, consequently, oral bioavailability. Using Trypanosoma brucei brucei cells as the parasite model and human normal kidney cells and mouse macrophage cells as the host model, we evaluated 30 new analogs synthesized through combinatorial chemistry. These analogs have fewer aromatic moieties and lower molecular weights than their predecessors. Several new analogs demonstrated IC50s in the low micromolar range, effectively inhibiting trypanosome cell growth without harming mammalian cells at the same concentration. We conducted a detailed structure-activity relationship (SAR) analysis and a docking study to assess the compounds' binding affinity to trypanosome tubulin homolog. The results revealed a correlation between binding energy and anti-Trypanosoma activity. Importantly, compound 7 displayed significant oral activity, effectively inhibiting trypanosome cell proliferation in mice.

VP5 protein of oncolytic herpes simplex virus type 2 induces apoptosis in A549 cells through TP53I3 protein.

Oncolytic virotherapy stands out as a burgeoning and promising therapeutic paradigm, harnessing the intrinsic cytotoxicity of oncolytic viruses for selective replication and dissemination within tumors. The primary mode of action revolves around the direct eradication of tumor cells. In our previous investigations, we formulated an oncolytic herpes simplex virus type 2 (OH2) and substantiated its anti-tumor efficacy both in vivo and in vitro. Subsequently, we embarked on a phase I/II clinical trial in China (NMPA, 2018L02743) and the USA (FDA, IND 27137) to assess OH2's safety, biodistribution, and anti-tumor activity as a standalone agent in patients with advanced solid tumors. In this investigation, our primary focus was to comprehend the influence of the major capsid protein VP5 of OH2 on its efficacy as an antitumor agent. Our findings underscore that the VP5 protein significantly amplifies OH2's oncolytic impact on A549 cells. Additionally, we observed that VP5 actively promotes the induction of apoptosis in A549 cells, both in vivo and in vitro. Through comprehensive transcriptional sequencing, we further authenticated that the VP5 protein triggers apoptosis-related signaling pathways and Gene Ontology (GO) terms in A549 cells. Moreover, we scrutinized differentially expressed genes in the p53-dependent apoptosis pathway and conducted meticulous in vitro validation of these genes. Subsequently, we delved deeper into unraveling the functional significance of the TP53I3 gene and conclusively affirmed that the VP5 protein induces apoptosis in A549 cells through the TP53I3 gene. These revelations illuminate the underlying mechanisms of OH2's antitumor activity and underscore the pivotal role played by the VP5 protein. The outcomes of our study harbor promising implications for the formulation of effective oncolytic virotherapy strategies in cancer treatment.

Microbiome in gynecologic malignancies: a bibliometric analysis from 2012 to 2022.

Microbiome and microbial dysbiosis have been proven to be involved in the carcinogenesis and treatment of gynecologic malignancies. However, there is a noticeable gap in the literature, as no comprehensive papers have covered general information, research status, and research frontiers in this field. This study addressed this gap by exploring the relationship between the gut and female reproductive tract (FRT) microbiome and gynecological cancers from a bibliometric perspective. Using VOSviewer 1.6.18, CiteSpace 6.1.R6, and HistCite Pro 2.1 software, we analyzed data retrieved from the Web of Science (WOS) Core Collection (WoSCC) database. Our dataset, consisting of 204 articles published from 2012 to 2022, revealed a consistent and upward publication trend. The United States and the United Kingdom were the primary driving forces, attributed to their prolificacy, high-quality output, and extensive cooperation. The University of Arizona Cancer Center, which is affiliated with the United States, ranked first among the top ten most prolific institutions. Frontiers in Cellular and Infection Microbiology emerged as the leading publisher. Herbst-Kralovetz MM led as the most productive author. Mitra A was the most influential author. Cervical cancer is notably associated with the microbiome, while endometrial and ovarian cancers are receiving increased attention in the last year. Intersections between the gut microbiome and estrogen are of growing importance. Current research focuses on identifying specific microbial species for etiological diagnosis, while frontiers mainly focus on the anticancer potential of microorganisms, such as regulating the effects of immune checkpoint inhibitors. In conclusion, this study sheds light on a novel and burgeoning direction of research, providing a one-stop overview of the microbiome in gynecologic malignancies. Its findings aim to help young researchers to identify research directions and future trends for ongoing investigations.

High resolution and large measurement range voltage sensing based on an optoelectronic oscillator utilizing an unbalanced Mach-Zehnder interferometer.

A voltage sensor with high resolution and large measurement range based on an optoelectronic oscillator (OEO) is proposed and experimentally demonstrated. The key component in the cavity to select the oscillating signal is a finite impulse response (FIR)-microwave photonic filter (MPF) which consists of a sinusoidal broadband optical signal, an unbalanced Mach-Zehnder interferometer (MZI), a section of dispersion compensating fiber, and a photodetector. The center frequency of the FIR-MPF is mainly determined by the free spectral range (FSR) of the FIR-MPF. In the lower arm of the MZI, a cylindrical piezoelectric ceramic (PZT) wrapped with a section of optical fiber acts as voltage sensing head. Due to the inverse piezoelectric effect of PZT, the variation of the voltage will cause radial deformation of the cylindrical PZT and then lead to the change of the FSR of the MZI, determining the shift of center frequency of FIR-MPF as well as the frequency of the oscillating signal of the OEO. Thus, by monitoring the shift of the oscillation frequency of the OEO using an electric spectrum analyzer or a digital signal processor, a high-speed interrogation and high-resolution voltage measurement can be realized. Additionally, in the proposed scheme, an infinite impulse response (IIR)-MPF consisting of a fiber ring resonator is cascaded with the FIR-MPF to ensure the single-mode oscillation of the OEO. The experimental results show that a total range of 1700 V voltage sensing from - 200 V to 1500 V is accomplished with the voltage sensitivity of 0.25 GHz/100 V and the resolution of 0.3 V. By adjusting the proportion of the length of single mode fiber between two branches of MZI, the impact of temperature can be greatly reduced. The proposed sensor offers advantages such as a large measurement range, high resolution, high-speed interrogation, and stability to temperature disturbances, making it highly suitable for sensing applications in smart grids.

A New and Practical Model of Human-like Ascending Aorta Aneurysm in Rats.

INTRODUCTION: Ascending aortic aneurysm is a serious health risk. In order to study ascending aortic aneurysms, elastase and calcium ion treatment for aneurysm formation are mainly used, but their aneurysm formation time is long, the aneurysm formation rate is low. Thus, this study aimed to construct a rat model of ascending aorta aneurysm with a short modeling time and high aneurysm formation rate, which may mimic the pathological processes of human ascending aorta aneurysm. METHODS: Cushion needles with different pipe diameters (1.0, 1.2, 1.4 and 1.6 mm) were used to establish a human-like rat model of ascending aortic aneurysm by narrowing the ascending aorta of rats and increasing the force of blood flow on the vessel wall. The vascular diameters were evaluated using color Doppler ultrasonography after two weeks. The characteristics of ascending aortic aneurysm in rats were detected by Masson's trichrome staining, Verhoeff's Van Gieson staining and hematoxylin and eosin staining while RT-PCR were utilized to assess the total RNA of cytokine interleukin-1ß, interleukin 6, transforming growth factor-beta1 and metalloproteinase 2. RESULTS: Two weeks after surgery, the ultrasound images and the statistical analysis demonstrated that the diameter of the ascending aorta in rats increased more than 1.5 times, similar to that in humans, indicating the success of animal modeling of ascending aortic aneurysm. Moreover, the optimal constriction diameter of the ascending aortic aneurysm model is 1.4 mm by the statistical analysis of the rate of ascending aortic aneurysm and mortality rate in rats with different constriction diameters. CONCLUSIONS: The human-like ascending aortic aneurysm model developed in this study can be used for the studies of the pathological processes and mechanisms in ascending aortic aneurysm in a more clinically relevant fashion.

Synthesis of polyoxometalate-pillared Zn-Cr layered double hydroxides for photocatalytic CO2 reduction and H2O oxidation.

Polyoxometalate (POM)-pillared Zn-Cr layered double hydroxides (LDHs) exhibited high photocatalytic activities in CO2 reduction and H2O oxidation reactions. For CO2 reduction in pure water, the CO production was 1.17 µmol g-1 after a 24 h reaction. For O2 evolution in NaIO3 solution, the O2 production reached 148.1 µmol g-1 after a 6 hour reaction. A mechanism study indicated that the electron transfer from Zn-Cr LDHs to POMs (SiW12O404-) promoted photocatalytic activities.

Elucidating the anti-inflammatory activity of platycodins in lung inflammation through pulmonary distribution dynamics and grey relational analysis of cytokines.

ETHNOPHARMACOLOGICAL RELEVANCE: Platycodonis Radix (PR) is a traditional herbal remedy used to prevent and treat lung inflammation, and platycodins are speculated to be the major active constituents. However, concrete experimental verification for this assertion remains absent thus far. AIM OF THE STUDY: This study aims to compare the pulmonary distribution dynamics of five platycodins and analyze their effects on cytokines. Through the grey relational analysis (GRA) between pulmonary active components and cytokines, the study ascertains platycodins as the potential effective component against lung inflammation. MATERIALS AND METHODS: A rat lung inflammation model was created using lipopolysaccharides (LPS). Pulmonary distribution dynamics were analyzed via LC-MS/MS. Cytokine changes and distribution patterns in lung tissues were studied by multi-factor reagent kit. GRA was applied to determine correlations between pulmonary components and cytokines. Finally, the anti-inflammatory properties of platycodins were further studied using LPS-induced BEAS-2B cells in vitro. RESULTS: The results showed that five platycodins (Platycodin D, Platycodin D3, Deapio Platycodin D, 3-O-ß-D-Glucopyranosyl Platycodigenin, and Platycodigenin) featured fast absorption rate, short time to peak, and slow metabolism rate. The pulmonary distribution dynamics were significantly affected within 2 h after LPS modeling. At the same time, PR altered the relationships among different cytokines induced by LPS stimulation, particularly inflammatory cytokines IL-6 and IFN-γ. The GRA results indicated good correlation between the pulmonary distribution dynamics of the five platycodins components and the changing patterns of cytokine levels, with Platycodin D3 contributing the most. Additionally, Platycodin D3 exhibited a protective role against LPS-induced inflammation by reducing the production of pro-inflammatory mediators such as IL-1ß, IL-8, and ROS, as well as increasing the expression of the anti-inflammatory mediator IL-10. CONCLUSIONS: Platycodins are the main anti-inflammatory agents in PR and there is a good correlation with cytokines. This contributes to the anti-pneumonia effect of PR.

The dual role of GoPGF reveals that the pigment glands are synthetic sites of gossypol in aerial parts of cotton.

Gossypol and the related terpenoids are stored in the pigment gland to protect cotton plants from biotic stresses, but little is known about the synthetic sites of these metabolites. Here, we showed that GoPGF, a key gene regulating gland formation, was expressed in gland cells and roots. The chromatin immunoprecipitation sequencing (ChIP-seq) analysis demonstrated that GoPGF targets GhJUB1 to regulate gland morphogenesis. RNA-sequencing (RNA-seq) showed high accumulation of gossypol biosynthetic genes in gland cells. Moreover, integrated analysis of the ChIP-seq and RNA-seq data revealed that GoPGF binds to the promoter of several gossypol biosynthetic genes. The cotton callus overexpressing GoPGF had dramatically increased the gossypol levels, indicating that GoPGF can directly activate the biosynthesis of gossypol. In addition, the gopgf mutant analysis revealed the existence of both GoPGF-dependent and -independent regulation of gossypol production in cotton roots. Our study revealed that the pigment glands are synthetic sites of gossypol in aerial parts of cotton and that GoPGF plays a dual role in regulating gland morphogenesis and gossypol biosynthesis. The study provides new insights for exploring the complex relationship between glands and the metabolites they store in cotton and other plant species.

Determination of multi-mycotoxins in vegetable oil via liquid chromatography-high resolution mass spectrometry assisted by a complementary liquid-liquid extraction.

The simultaneous determination of multi-mycotoxins in food commodities are highly desirable due to their potential toxic effects and mass consumption of foods. Herein, liquid chromatography-quadrupole exactive orbitrap mass spectrometry was proposed to analyze multi-mycotoxins in commercial vegetable oils. Specifically, the method featured a successive liquid-liquid extraction process, in which the complementary solvents consisted of acetonitrile and water were optimized. Resultantly, matrix effects were reduced greatly. External calibration approach revealed good quantification property for each analyte. Under optimal conditions, the recovery ranging from 80.8% to 109.7%, relative standard deviation less than 11.7%, and good limit of quantification (0.35 to 45.4 ng/g) were achieved. The high accuracy of proposed method was also validated. The detection of 20 commercial vegetable oils revealed that aflatoxins B1 and B2, zearalenone were observed in 10 real samples. The as-developed method is simple and low-cost, which merits the wide applications for scanning mycotoxins in oil matrices.

Pharmacokinetics and Tissue Distribution of Isovitexin-2''-O-ß-D-glucopyranoside (IVG) in Sprague-Dawley Rats.

BACKGROUND: Isovitexin-2"-O-D-glucopyranoside (IVG) has been known to exhibit sedative and hypnotic effects. However, there is little understanding of the in vivo pharmacokinetics and tissue distribution of IVG. OBJECTIVE: This study aimed to investigate the pharmacokinetics and tissue distribution of IVG. METHODS: The study employed an HPLC-ESI-MS/MS method to analyze the pharmacokinetics and tissue distribution of IVG. RESULTS: Under mass spectrometry, IVG and internal standard (IS) showed strong negative ionization signals. MRM analysis chose ion transitions m/z 593.3 → 293.0 (IVG) and m/z 579.8 → 271.4 (IS). Method validation indicated high precision, accuracy, and reliability with a quantitation limit under 20 ng/mL. After intravenously administering 5.0 mg/kg of IVG, rapid clearance from rat plasma was observed, with a half-life (t1/2) of 3.49 ± 0.99 h and a clearance rate of 54.53 ± 11.90 mL/kg/h. The area under the curve (AUC0-12h) of 37.79 ± 7.65 µg·h/mL indicated a brisk metabolic rate. Evaluating the tissue distribution, the highest accumulation was seen in the liver (30.32 ± 3.06 µg/g), followed by the kidney (20.58 ± 2.12 µg/g) and intestine (6.69 ± 0.93 µg/g), suggesting a propensity for IVG to concentrate in these tissues. Importantly, the presence of IVG in the brain underlines its potential to traverse the blood-brain barrier. These findings revealed that following intravenous administration, IVG was swiftly and broadly distributed throughout various rat tissues. CONCLUSION: This study provides valuable information on the pharmacokinetics and tissue distribution of IVG, implicating its potential as a novel and effective drug candidate for sedative and anxiolytic treatment.

Immunomodulatory effect of Ti-Cu alloy by surface nanostructure synergistic with Cu2+ release.

The inflammatory response induced by implant/macrophage interaction has been considered to be one of the vital factors in determining the success of implantation. In this study, TiCuNxOy coating with an immunomodulatory strategy was proposed for the first time, using nanostructured TiCuNxOy coating synthesized on Ti-Cu alloy by oxygen and nitrogen plasma-based surface modification. It was found that TiCuNxOy coating inhibited macrophage proliferation but stimulated macrophage preferential activation and presented an elongated morphology due to the surface nanostructure. The most encouraging discovery was that TiCuNxOy coating promoted the initial pro-inflammatory response of macrophages and then accelerated the M1-to-M2 transition of macrophages via a synergistic effect of fast-to-slow Cu2+ release and surface nanostructure, which was considered to contribute to initial infection elimination and tissue healing. As expected, TiCuNxOy coating released desirable Cu2+ and generated a favorable immune response that facilitated HUVEC recruitment to the coating, and accelerated proliferation, VEGF secretion and NO production of HUVECs. On the other hand, it is satisfying that TiCuNxOy coating maintained perfect long-term antibacterial activity (≥99.9%), mainly relying on Cu2O/CuO contact sterilization. These results indicated that TiCuNxOy coating might offer novel insights into the creation of a surface with immunomodulatory effects and long-term bactericidal potential for cardiovascular applications.

Optimization of ultrasonic-assisted extraction of Platycodon grandiflorum polysaccharides and evaluation of its structural, antioxidant and hypoglycemic activity.

The study aimed to improve the extraction rate of Platycodon grandiflorum roots polysaccharides (PGPs) using ultrasound-assisted extraction (UAE). A comparative analysis was undertaken to evaluate polysaccharides content, molecular weight distribution, monosaccharide composition, preliminary structure, antioxidant, and hypoglycemic activity of UAE in comparison with heating water extraction (HWE). The optimum extraction conditions included a liquid-to-material ratio of 20 mL/g, ultrasonic power of 150 W, extraction temperature of 70 ℃, and extraction time of 20 min, resulting in a significantly greater polysaccharides (12.011 ± 0.91 %) compared to HWE (7.62 ± 0.18 %). Through Sephacryl S-100 column elution, two homogenous fraction (PGP-U extracted with UAE and PGP-H extracted with HAE) were obtained. The molecular weight of PGP-U and PGP-H was 3.14 kDa and 3.44 kDa, respectively, mainly composed of different proportions of fourteen monosaccharides. Fourier transform infrared spectroscopy (FT-IR) and Nuclear Magnetic Resonance (NMR) spectra experiment results showed that the two polysaccharides were pyranose ring with α- and ß-glycoside bond. PGP-U and PGP-H exhibited specific antioxidant activities, encompassing total reducing force, scavenging of DPPH radicals, ABTs radicals and hydroxyl radicals in vitro, along with mitigation of H2O2-induced damage in HepG2 cells. Moreover, PGP-U exerted significantly stronger inhibitory activities against α-amylase and α-glucosidase and could significantly enhances the glucose uptake capacity and intracellular glycogen content of insulin-resistant HepG2 (IR-HepG2) cells.

Silicon Combined with Melatonin Reduces Cd Absorption and Translocation in Maize.

Cadmium (Cd) is one of the most toxic and widely distributed heavy metal pollutants, posing a huge threat to crop production, food security, and human health. Corn is an important food source and feed crop. Corn growth is subject to Cd stress; thus, reducing cadmium stress, absorption, and transportation is of great significance for achieving high yields, a high efficiency, and sustainable and safe corn production. The use of silicon or melatonin alone can reduce cadmium accumulation and toxicity in plants, but it is unclear whether the combination of silicon and melatonin can further reduce the damage caused by cadmium. Therefore, pot experiments were conducted to study the effects of melatonin and silicon on maize growth and cadmium accumulation. The results showed that cadmium stress significantly inhibited the growth of maize, disrupted its physiological processes, and led to cadmium accumulation in plants. Compared to the single treatment of silicon or melatonin, the combined application of melatonin and silicon significantly alleviated the inhibition of the growth of maize seedlings caused by cadmium stress. This was demonstrated by the increased plant heights, stem diameters, and characteristic root parameters and the bioaccumulation in maize seedlings. Under cadmium stress, the combined application of silicon and melatonin increased the plant height and stem diameter by 17.03% and 59.33%, respectively, and increased the total leaf area by 43.98%. The promotion of corn growth is related to the reduced oxidative damage under cadmium stress, manifested in decreases in the malondialdehyde content and relative conductivity and increases in antioxidant enzyme superoxide dismutase and guaiacol peroxidase activities, as well as in soluble protein and chlorophyll contents. In addition, cadmium accumulation in different parts of maize seedlings and the health risk index of cadmium were significantly reduced, reaching 48.44% (leaves), 19.15% (roots), and 20.86% (health risk index), respectively. Therefore, melatonin and silicon have a significant synergistic effect in inhibiting cadmium absorption and reducing the adverse effects of cadmium toxicity.

Negative pressure wound therapy promotes wound healing of diabetic foot ulcers by up-regulating PRDX2 in wound margin tissue.

To understand the changes in the peroxiredoxin-2 (PRDX2) expression level in the wound margin tissue (T-PRDX2) of patients with diabetic foot ulcer (DFU) before and after negative pressure wound therapy (NPWT). Additionally, the study aimed to explore the association between PRDX2 expression and the treatment outcome of DFUs to provide a new theoretical basis for revealing the mechanism of NPWT promoting the healing of DFUs. Fifty-six type 2 diabetes patients with foot ulcers undergoing NPWT (the DFU group) and 28 patients with chronic lower limb skin ulcers with normal glucose tolerance undergoing NPWT (the skin ulcer control [SUC] group) were included in the study. T-PRDX2 was detected using Western blotting, and the superoxide dismutase (SOD) activity and the malondialdehyde (MDA) and glutathione (GSH) levels were detected using a biochemical method. In addition, in vitro experiments were conducted to determine the effect of PRDX2 expression on normal human dermal fibroblast (NHDF) proliferation, migration, and apoptosis. Before NPWT, the DFU group exhibited a significantly lower T-PRDX2 expression level compared with the SUC group. After one week of NPWT, the T-PRDX2 expression level, SOD activity, and GSH content in the wound margin tissues of the DFU and SUC groups significantly increased compared with the before NPWT levels. Conversely, the inflammatory indicators (white blood cell, neutrophil percentage, C-reactive protein, and procalcitonin) and MDA content were significantly lower than the before NPWT levels. The expression changes of T-PRDX2 before and after NPWT in the DFU and SUC groups were positively correlated with the 4-week wound healing rate. In vitro experiments demonstrated that PRDX2 could alleviate the oxidative stress in NHDFs, thereby promoting their proliferation and migration, while reducing cell apoptosis. NPWT promotes DFU healing by increasing T-PRDX2, and changes in the T-PRDX2 might be associated with the therapeutic effect of NPWT.

Genome-Wide Identification, Phylogeny and Expression Analysis of Subtilisin (SBT) Gene Family under Wheat Biotic and Abiotic Stress.

The subtilisin-like protease (SBT) family is widely known for its role in stress resistance to a number of stressors in different plant species, but is rarely studied in wheat. Subtilisin-like serine proteases (SBTs) are serine proteolytic enzymes that hydrolyze proteins into small peptides, which bind to receptors as signal molecules or ligands and participate in signal transduction. In this study, we identified 255 putative SBT genes from the wheat reference genome and then divided these into seven clades. Subsequently, we performed syntenic relation analysis, exon-intron organization, motif composition, and cis-element analysis. Further, expression analysis based on RNA-seq and tissue-specific expression patterns revealed that TaSBT gene family expression has multiple intrinsic functions during various abiotic and biotic stresses. Analysis of RNA-seq expression assays and further validation through qRT PCR suggested that some of the TaSBT genes have significant changes in expression levels during Pst interaction. TaSBT7, TaSBT26, TaSBT102, and TaSBT193 genes showed increasing expression levels during compatible and non-compatible interactions, while the expression levels of TaSBT111 and TaSBT213 showed a decreasing trend, indicating that these members of the wheat SBT gene family may have a role in wheat's defense against pathogens. In conclusion, these results expand our understanding of the SBT gene family, and provide a valuable reference for future research on the stress resistance function and comprehensive data of wheat SBT members.

Role of Increased miR-222-3p Expression in Peripheral Blood and Wound Marginal Tissues of Type 2 Diabetes Mellitus Patients with Diabetic Foot Ulcer.

Purpose: To study the correlations of miR-222-3p expression in the peripheral blood and wound marginal tissues of type 2 diabetes mellitus (T2DM) patients with the onset of diabetic foot ulcer (DFU), as well as explore the clinical value possessed by miR-222-3p in the diagnosis and treatment outcomes of DFU. Methods: The study included 70 T2DM patients who did not suffer foot ulcers (T2DM group), 146 T2DM patients who suffered foot ulcers (DFU group), as well as 70 normal controls (NC group). Quantitative real-time PCR determined the MiR-222-3p relative expression. Clinical features and risk factors regarding DFU were assessed. Multiple stepwise logistic regression analysis assisted in confirming whether miR-222-3p expression could serve for independently predicting the risk factors for DFU. ROC curve analysis evaluated the diagnostic value exhibited by miR-222-3p level against DFU. Results: T2DM group exhibited an obviously higher MiR-222-3p expression relative to NC group [1.98 (0.98, 3.62) vs 0.92 (0.61, 1.87)] (P < 0.01), but DFU group exhibited an obviously higher miR-222-3p expression relative to T2DM group [5.61 (1.98, 10.24) vs 1.98 (0.98, 3.62)] (P < 0.01). Besides, miR-222-3p expression presented a negative correlation with DFU healing rate (P < 0.05). According to Kaplan-Meier survival curve analysis, the group with high miR-222-3p expression showed higher unhealed DFU cumulative rate relative to the group with low expression (log-rank, P = 0.011, 0.001, respectively). Multivariate logistic regression analysis confirmed that high miR-222-3p expressions could independently predict DFU risk (OR=3.85, 95% CI 1.18~12.37, P = 0.008). According to the ROC curve analysis, the AUC of miR-222-3p specific to DFU diagnosis reached 0.803, with the best sensitivity of 95.93% and best specificity of 96.27%. Conclusion: The increased expression of miR-222-3p in the peripheral blood of T2DM patients is closely related to the occurrence of DFU. MiR-222-3p is a biomarker with potential clinical value in diagnosing and evaluating the prognosis of DFU.

Mechanism of wound repair in diabetic rats using nanosilver-free alginate dressing.

OBJECTIVE: Nanosilver-alginate dressing can effectively promote the healing of diabetic wounds in rats. However, due to the potential toxicity of nanosilver, its widespread application in hard-to-heal wound healing is limited. In the present study, the role and potential mechanism of nanosilver-free alginate gel (NSFAG) in the healing process of diabetic wounds were explored. METHOD: A diabetic rat skin wound model was established, and wounds were treated with saline (NC group), nanosilver gel (NSG group) or nanosilver-free alginate gel (NSFAG group) for seven consecutive days. RESULTS: NSFAG significantly promoted wound healing and increased the content of protein and hydroxyproline in granulation tissues, and was superior to NSG (p<0.05). Immunohistochemical analyses revealed that the skin wound tissue structure of the NSFAG group was intact, and the number of skin appendages in the dermis layer was significantly higher compared with the NC group and the NSG group (p<0.05). Western blot analysis found that the protein expression of the epidermal stem cell marker molecules CK19 and CK14 as well the proliferation marker of keratinocytes Ki67 in the NSFAG group was significantly higher compared with the NC group or NSG group (p<0.05). Additionally, the proliferation marker of keratinocytes Ki67 in the NSFAG group was significantly higher compared with the NC or NSG group (p<0.05). Immunofluorescence staining analyses indicated that the CK19- and CK14-positive cells were mainly distributed around the epidermis and the newly formed appendages in the NSFAG group, and this result was not observed in the NC or NSG groups. CONCLUSION: The present findings demonstrate that NSFAG can effectively accelerate wound healing in diabetic rats by promoting epidermal stem cell proliferation and differentiation into skin cells, as well as formation of granulation tissue, suggesting that it can be a potential dressing for diabetic wounds.