Pathogen detection in suspected spinal infection: metagenomic next-generation sequencing versus culture.

PURPOSE: The aim is to compare the pathogen detection performance of metagenomic next-generation sequencing (mNGS) and the culturing of percutaneous needle biopsy samples obtained from an individual with a suspected spinal infection. METHODS: A retrospective study of 141 individuals with a suspected spinal infection was conducted, and mNGS was performed. The microbial spectra and detection performance between mNGS and the culturing-based method were compared, and the effects of antibiotic intervention and biopsy on the detection performance were assessed. RESULTS: The microorganisms isolated most commonly via the culturing-based method were Mycobacterium tuberculosis (n = 21), followed by Staphylococcus epidermidis (n = 13). The most common microorganisms detected via mNGS were Mycobacterium tuberculosis complex (MTBC) (n = 39), followed by Staphylococcus aureus (n = 15). The difference in the type of detected microorganisms between culturing and mNGS was observed only in Mycobacterium (P = 0.001). mNGS helped identify potential pathogens in 80.9% of cases, which was significantly higher than the positivity rate of 59.6% observed for the culturing-based method (P < 0.001). Moreover, mNGS had a sensitivity of 85.7% (95% CI, 78.4% to 91.3%), a specificity of 86.7% (95% CI, 59.5% to 98.3%), and sensitivity gains of 35% (85.7% vs. 50.8%; P < 0.001) during culturing, while no differences were observed in the specificity (86.7% vs. 93.3%; P = 0.543). In addition, antibiotic interventions significantly lowered the positivity rate of the culturing-based method (66.0% vs. 45.5%, P = 0.021) but had no effects on the results of mNGS (82.5% vs. 77.3%, P = 0.467). CONCLUSION: The use of mNGS could result in a higher detection rate compared to that observed with the culturing-based method in an individual with spinal infection and is particularly valuable for evaluating the effects of a mycobacterial infection or previous antibiotic intervention.

Exosomes-loaded electroconductive nerve dressing for nerve regeneration and pain relief against diabetic peripheral nerve injury.

Over the years, electroconductive hydrogels (ECHs) have been extensively applied for stimulating nerve regeneration and restoring locomotor function after peripheral nerve injury (PNI) with diabetes, given their favorable mechanical and electrical properties identical to endogenous nerve tissue. Nevertheless, PNI causes the loss of locomotor function and inflammatory pain, especially in diabetic patients. It has been established that bone marrow stem cells-derived exosomes (BMSCs-Exos) have analgesic, anti-inflammatory and tissue regeneration properties. Herein, we designed an ECH loaded with BMSCs-Exos (ECH-Exos) electroconductive nerve dressing to treat diabetic PNI to achieve functional recovery and pain relief. Given its potent adhesive and self-healing properties, this laminar dressing is convenient for the treatment of damaged nerve fibers by automatically wrapping around them to form a size-matched tube-like structure, avoiding the cumbersome implantation process. Our in vitro studies showed that ECH-Exos could facilitate the attachment and migration of Schwann cells. Meanwhile, Exos in this system could modulate M2 macrophage polarization via the NF-κB pathway, thereby attenuating inflammatory pain in diabetic PNI. Additionally, ECH-Exos enhanced myelinated axonal regeneration via the MEK/ERK pathway in vitro and in vivo, consequently ameliorating muscle denervation atrophy and further promoting functional restoration. Our findings suggest that the ECH-Exos system has huge prospects for nerve regeneration, functional restoration and pain relief in patients with diabetic PNI.

Triple-functional bone adhesive with enhanced internal fixation, bacteriostasis and osteoinductive properties for open fracture repair.

At present, effective fixation and anti-infection implant materials represent the mainstay for the treatment of open fractures. However, external fixation can cause nail tract infections and is ineffective for fixing small fracture fragments. Moreover, closed reduction and internal fixation during the early stage of injury can lead to potential bone infection, conducive to bone nonunion and delayed healing. Herein, we designed a bone adhesive with anti-infection, osteogenic and bone adhesion fixation properties to promote reduction and fixation of open fractures and subsequent soft tissue repair. It was prepared by the reaction of gelatin (Gel) and oxidized starch (OS) with vancomycin (VAN)-loaded mesoporous bioactive glass nanoparticles (MBGNs) covalently cross-linked with Schiff bases. Characterization and adhesion experiments were conducted to validate the successful preparation of the Gel-OS/VAN@MBGNs (GOVM-gel) adhesive. Meanwhile, in vitro cell experiments demonstrated its good antibacterial effects with the ability to stimulate bone marrow mesenchymal stem cell (BMSCs) proliferation, upregulate the expression of alkaline phosphatase (ALP) and osteogenic proteins (RunX2 and OPN) and enhance the deposition of calcium nodules. Additionally, we established a rat skull fracture model and a subcutaneous infection model. The histological analysis showed that bone adhesive enhanced osteogenesis, and in vivo experiments demonstrated that the number of inflammatory cells and bacteria was significantly reduced. Overall, the adhesive could promote early reduction of fractures and antibacterial and osteogenic effects, providing the foothold for treatment of this patient population.

DNA methylation variation is crucial to restore adventitious rooting ability during in vitro shoot culture-induced rejuvenation in apple rootstock.

In vitro shoot culture has been widely used for restoring adventitious rooting ability in rooting recalcitrant woody perennial species for the past few decades, but its molecular mechanism is largely uncovered. DNA methylation is an essential epigenetic mark that participates in many biological processes. Recent reports suggested a role of DNA methylation in vitro culture in plants. In this study, we characterized the single-base resolution DNA methylome and transcriptome of adult and in vitro shoot culture-induced rejuvenation cuttings of apple rootstock M9T337. We found a global decrease in DNA methylation during rejuvenation, which may be correlated with increased expression of DNA demethylase genes and decreased expression of DNA methyltransferase genes. We additionally documented DNA hypomethylation in 'T337'_R in gene protomer associated with higher transcript levels of several adventitious rooting-related genes. The application of a DNA methylation inhibitor (5-azacytidine) enhanced the adventitious rooting ability and the expression level of adventitious rooting-related genes, such as, MdANT, MdMPK3, MdABCB21, MdCDC48, MdKIN8B, pri-MdMIR156a5 and pri-MdMIR156a12. Together, the DNA hypomethylation is critical for the rejuvenation-dependent adventitious rooting ability in apple rootstock. In addition, increased DNA methylation was also found in thousands of genes in 'T337'_R. We additionally documented that DNA hypermethylation is required for inhibition of adventitious rooting-repressed genes, such as MdGAD5a, encoding glutamate decarboxylase, which can catalyze glutamate decarboxylated to form γ-aminobutyric acid (GABA). Our results revealed that in vitro shoot culture-dependent DNA methylation variation plays important roles in adventitious rooting in apple rootstock.

Single-cell profiling of healthy human kidney reveals features of sex-based transcriptional programs and tissue-specific immunity.

Knowledge of the transcriptional programs underpinning the functions of human kidney cell populations at homeostasis is limited. We present a single-cell perspective of healthy human kidney from 19 living donors, with equal contribution from males and females, profiling the transcriptome of 27677 cells to map human kidney at high resolution. Sex-based differences in gene expression within proximal tubular cells were observed, specifically, increased anti-oxidant metallothionein genes in females and aerobic metabolism-related genes in males. Functional differences in metabolism were confirmed in proximal tubular cells, with male cells exhibiting higher oxidative phosphorylation and higher levels of energy precursor metabolites. We identified kidney-specific lymphocyte populations with unique transcriptional profiles indicative of kidney-adapted functions. Significant heterogeneity in myeloid cells was observed, with a MRC1+LYVE1+FOLR2+C1QC+ population representing a predominant population in healthy kidney. This study provides a detailed cellular map of healthy human kidney, and explores the complexity of parenchymal and kidney-resident immune cells.

Facile Fabrication of 3D-Printed Porous Ti6Al4V Scaffolds with a Sr-CaP Coating for Bone Regeneration.

To improve osseointegration caused by the stress-shielding effect and the inert nature of titanium-based alloys, in this work, we successfully constructed a strontium calcium phosphate (Sr-CaP) coating on three-dimensional (3D)-printed Ti6Al4V scaffolds to address this issue. The energy-dispersive X-ray spectroscopy (EDS) and X-ray diffraction (XRD) results indicated that the coatings with and without Sr doping mainly consisted of CaHPO4. The bonding strength of Sr doping coating met the required ISO 13 779-4-2018 standard (≥15 MPa). The in vitro results suggested that the Sr-CaP-modified Ti6Al4V scaffolds were found to effectively promote mice bone-marrow stem cell (mBMSC) adhesion, spreading, and osteogenesis. The in vivo experiments also showed that the Sr-CaP-modified Ti6Al4V scaffolds could significantly improve bone regeneration and osseointegration. More importantly, Sr-doped CaP-coated Ti6Al4V scaffolds were found to accelerate bone healing in comparison to CaP-coated Ti6Al4V scaffolds. The Sr-CaP-modified Ti6Al4V scaffolds are considered a promising strategy to develop bioactive surfaces for enhancing the osseointegration between the implant and bone tissue.

In Vitro Study of Degradation and Cytocompatibility of Ceramics/PLA Composite Coating on Pure Zinc for Orthopedic Application.

Zinc and its alloys are considered to be next-generation materials for fabricating absorbable biomedical devices. However, cytotoxicity has been reported to be associated with rapid degradation. To address these issues, a composite coating (PLA/Li-OCP) consisting of CaHPO4 conversion coating (Ca-P) and polylactic acid (PLA) decorated with Li-octacalcium phosphate particles was constructed on pure zinc. The immersion tests showed that the presence of Ca-P coating and PLA/Li-OCP coating on pure zinc could reduce the pH value. Compared with Ca-P coating, the introduction of the PLA/Li-OCP film on the Ca-P-coated samples could enhance the corrosion resistance, and there was one order of magnitude decrease in the corrosion current density. The cytocompatibility assay suggested that the PLA/Li-OCP coating favored the cell viability and upregulated the expression of related osteogenic-genes including RUNX2, OCN, and BMP. Therefore, the presence of the PLA/Li-OCP coating on pure zinc could effectively improve the degradation rate and cytocompatibility of pure zinc.

Comprehensive Genome-Wide Identification and Transcript Profiling of GABA Pathway Gene Family in Apple (Malus domestica).

γ-Aminobutyric Acid (GABA), a four-carbon non-protein amino acid, is a significant component of the free amino acid pool in most prokaryotic and eukaryotic organisms. GABA is involved in pH regulation, maintaining C/N balance, plant development and defence, as well as a compatible osmolyte and an alternative pathway for glutamate utilization via anion flux. Glutamate decarboxylase (GAD, EC 4.1.1.15) and GABA transaminase (GABA-T, EC 2.6.1.19) are two key enzymes involved in the synthesis and metabolism of GABA. Recently, GABA transporters (GATs), protein and aluminium-activated malate transporter (ALMT) proteins which function as GABA receptors, have been shown to be involved in GABA regulation. However, there is no report on the characterization of apple GABA pathway genes. In this study, we performed a genome-wide analysis and expression profiling of the GABA pathway gene family in the apple genome. A total of 24 genes were identified including five GAD genes (namely MdGAD 1-5), two GABA-T genes (namely MdGABA-T 1,2), 10 GAT genes (namely GAT 1-10) and seven ALMT genes (namely MdALMT1-7). These genes were randomly distributed on 12 chromosomes. Phylogenetic analyses grouped GABA shunt genes into three clusters-cluster I, cluster II, and cluster III-which had three, four, and five genes, respectively. The expression profile analysis revealed significant MdGAD4 expression levels in both fruit and flower organs, except pollen. However, there were no significant differences in the expression of other GABA shunt genes in different tissues. This work provides the first characterization of the GABA shunt gene family in apple and suggests their importance in apple response to abiotic stress. These results can serve as a guide for future studies on the understanding and functional characterization of these gene families.

Genome-wide identification and characterization of Respiratory Burst Oxidase Homolog genes in six Rosaceae species and an analysis of their effects on adventitious rooting in apple.

Adventitious root formation is essential for plant propagation, development, and response to various stresses. Reactive oxygen species (ROS) are essential for adventitious root formation. However, information on Respiratory Burst Oxidase Homolog (RBOH), a key enzyme that catalyzes the production ROS, remains limited in woody plants. Here, a total of 44 RBOH genes were identified from six Rosaceae species (Malus domestica, Prunus avium, Prunus dulcis 'Texas', Rubus occidentalis, Fragaria vesca and Rosa chinensis), including ten from M. domestica. Their phylogenetic relationships, conserved motifs and gene structures were analyzed. Exogenous treatment with the RBOH protein inhibitor diphenyleneiodonium (DPI) completely inhibited adventitious root formation, whereas exogenous H2O2 treatment enhanced adventitious root formation. In addition, we found that ROS accumulated during adventitious root primordium inducing process. The expression levels of MdRBOH-H, MdRBOH-J, MdRBOH-A, MdRBOH-E1 and MdRBOH-K increased more than two-fold at days 3 or 9 after auxin treatment. In addition, cis-acting element analysis revealed that the MdRBOH-E1 promoter contained an auxin-responsive element and the MdRBOH-K promoter contained a meristem expression element. Based on the combined results from exogenous DPI and H2O2 treatment, spatiotemporal expression profiling, and cis-element analysis, MdRBOH-E1 and MdRBOH-K appear to be candidates for the control of adventitious rooting in apple.

[Posterior spinal canal decompression with screw fixation and reconstruction of three vertebral column for thoracolumbar burst fractures complicated with nerve injury].

OBJECTIVE: To explore the feasibility and clinical effect of posterior spinal canal decompression with pedicle screw fixation and reconstruction of anterior and middle vertebral column for thoracolumbar burst fractures complicated with nerve injury. METHODS: A total of 36 patients with thoracolumbar burst fractures treated from March 2011 to April 2016 were enrolled in the retrospective study. There were 20 males and 16 females, aged from 21 to 52 years old with an average of 37.6 years. All the fractures were located on a single segment, 8 cases of T1111, 10 cases of T12, 12 cases of L1, 6 cases of L2. According to thoracolumbar injury classification and severity score(TLICS) system, the score was 7 to 9 points, with an average of 7.4 points. According to the America Spine Injury Association(ASIA) grade, 4 cases were type A, 10 cases were type B, 14 cases were type C, 8 cases were type D. All the patients underwent posterior spinal canal decompression with pedicle screw fixation and reconstruction of anterior and middle vertebral column. The recovery of nerve function was evaluated by ASIA grading. The correction of kyphosis(Cobb angle), the volume change of injuried spinal canal, the change of anterior border height of injury vertebra which can be observed by X-rays;the internal fixation loosening and breakage and all the information of bone fusion were recorded. RESULTS: All the operations were successful, the mean operative time and intraoperative blood loss were(2.8±0.3) h (2.1 to 3.5 h) and (880±120) ml(550 to 1 350 ml), respectively. All the incisions got primary healing. All the patients were followed up for 12 to 28 months with an average of 18.4 months. All the patients obtained satisfactory bone fusion. No pseudoarticulation formation was found, and there was no loosening, breakage of pedicle screws or displacement of titanium mesh. Neurological function was improved in different degree, except in one patient with grade A and another one with grade B. According to the ASIA grade, there were 1 case of type A, 1 case of type B, 7 cases of type C, 10 cases of type D and 17 cases of type E, postoperatively. At 3 days after operative, the anterior border height of injury vertebra, Cobb angle and the volume changes of injury spinal canal were obviously improved(P<0.05), and there was no significant difference between postoperative at 3 days and final follow-up(P>0.05). CONCLUSIONS: Spinal canal decompression with screw fixation and reconstruction of anterior and middle vertebral column through posterior midline approach is a safe and effective method in the treatment of thoracolumbar burst fractures with nerve injury, it is worthy to be popularized. It can complete the spinal canal decompression of 360 degree, reduction of fractures and reconstruction of vertebral three-column at the same time through a single posterior approach. The advantages includes less trauma, perfect decompression, good stability and etc.

The Exploration of Health-Related Quality of Life.

The purpose of this study was to explore quality of life (QoL) and the factors influencing QoL in gynecological cancer patients. One hundred sixty-seven patients with gynecologic cancers were recruited from a district hospital in Southern Taiwan. The instruments used included the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core-30 Version 3.0 in Chinese (EORTC QLQ-C30), the Symptom Distress Scale, and demographic characteristics and disease-related variables. The results showed that the mean score for the QLQ-C30 was 61.13 ( SD = 22.71). In the stepwise regression model, two factors predicted overall global QoL: symptom distress (33.8%) and current occupation (2.2%). These predictors accounted for 36.0% of the total variance. These results showed that symptom distress was a predictor of QoL. This study provides a reference for use when designing improved educational care programs that reduce patient symptom distress and enhance gynecologic cancer patients' QoL.

Tissue factor-containing microparticles released from mesangial cells in response to high glucose and AGE induce tube formation in microvascular cells.

Hyperglycaemia and the associated formation of advanced glycation end-products (AGE) have been implicated in the pathogenesis of diabetic vasculopathy. In addition to its role in coagulation, tissue factor (TF) is known to regulate vascular proliferation and angiogenesis. In this study, the influence of AGE and glucose on the expression of TF in human renal mesangial cells (HRMC) and the subsequent induction of capillary formation by human dermal microvascular endothelial cells (HDMEC) were measured. Furthermore, the activity of TF, incorporated into microparticles was investigated. Both AGE and elevated glucose were capable of upregulating the expression of TF expression in a concentration-dependent manner in HRMC but not in HDMEC. This TF antigen and activity in the conditioned media from HRMC was associated with microparticles. Moreover, the formation of capillaries was readily induced on supplementation of HDMEC with conditioned media, from AGE-treated or high glucose-treated HRMC but not on incubation of HDMEC with either AGE or hyperphysiological concentrations of glucose. Furthermore, the rate of capillary formation was suppressed on incubation of the conditioned media with a polyclonal antibody against TF but not against VEGF. This study indicates that TF-containing microparticles are an important pro-inflammatory mediator acting as a mediator between elevated glucose and the development of diabetic vasculopathy by altering the angiogenic properties of endothelial cells and offers one explanation for the correlation between diabetes and microvascular disease.

Differential gene expression profiling in HELLP syndrome placentas.

The aim of this study was to identify differentially expressed genes by suppression subtractive hybridization (SSH) in HELLP placentas. Two cDNA libraries were constructed; HSI (HELLP subtracted induced or upregulated) and HSS (HELLP subtracted suppressed or downregulated). Two hundred eighty-eight cDNA clones were sequenced; 37 were matched to GenBank entries and included genes in cell communication and organization, cellular processes, genetic information processing, and metabolic processes. A subgroup of 11 genes of interest was further selected for real-time quantitative polymerase chain reaction confirmation. Results showed no differences in expression of chosen upregulated genes between HELLP and non-HELLP placentas; 6 HELLP downregulated genes were significantly suppressed. Two genes related to production of secreted proteins, CTHRC1 and SERPINE2. SERPINE2 (PAI-1) is a soluble protease inhibitor and is a potential biomarker by Western blot analysis, and the protein is significantly decreased in HELLP placentas. SERPINE2 might be tested clinically in patients for early diagnosis of HELLP syndrome.