Ehrlichia chaffeensis Etf-3 Induces Host RAB15 Upregulation for Bacterial Intracellular Growth.

Ehrlichia chaffeensis infects human monocytes or macrophages and causes human monocytic ehrlichiosis (HME), an emerging life-threatening zoonosis. After internalization, E. chaffeensis resides in membrane-bound inclusions, E. chaffeensis-containing vesicles (ECVs), which have early endosome-like characteristics and fuse with early autophagosomes but not lysosomes, to evade host innate immune microbicidal mechanisms and obtain nutrients for bacterial intracellular growth. The mechanisms exploited by E. chaffeensis to modulate intracellular vesicle trafficking in host cells have not been comprehensively studied. Here, we demonstrate that E. chaffeensis type IV secretion system (T4SS) effector Etf-3 induces RAB15 upregulation in host cells and that RAB15, which is localized on ECVs, inhibits ECV fusion with lysosomes and induces autophagy. We found that E. chaffeensis infection upregulated RAB15 expression using qRT-PCR, and RAB15 was colocalized with E. chaffeensis using confocal microscopy. Silence of RAB15 using siRNA enhanced ECV maturation to late endosomes and fusion with lysosomes, as well as inhibited host cell autophagy. Overexpression of Etf-3 in host cells specifically induced RAB15 upregulation and autophagy. Our findings deepen the understanding of E. chaffeensis pathogenesis and adaptation in hosts as well as the function of RAB15 and facilitate the development of new therapeutics for HME.

scGPT: toward building a foundation model for single-cell multi-omics using generative AI.

Generative pretrained models have achieved remarkable success in various domains such as language and computer vision. Specifically, the combination of large-scale diverse datasets and pretrained transformers has emerged as a promising approach for developing foundation models. Drawing parallels between language and cellular biology (in which texts comprise words; similarly, cells are defined by genes), our study probes the applicability of foundation models to advance cellular biology and genetic research. Using burgeoning single-cell sequencing data, we have constructed a foundation model for single-cell biology, scGPT, based on a generative pretrained transformer across a repository of over 33 million cells. Our findings illustrate that scGPT effectively distills critical biological insights concerning genes and cells. Through further adaptation of transfer learning, scGPT can be optimized to achieve superior performance across diverse downstream applications. This includes tasks such as cell type annotation, multi-batch integration, multi-omic integration, perturbation response prediction and gene network inference.

A machine learning model for identifying systemic lupus erythematosus through laboratory information system and electronic medical record.

OBJECTIVES: Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease. Its diagnosis poses significant challenges especially at early stages and in atypical cases. The aim of this study was to develop a machine learning model based on common laboratory tests that can aid SLE diagnosis. METHODS: A standard protocol was developed to collect data of SLE and control immune diseases. A 10-fold cross-validation was performed in the modeling dataset (n=862), and an external dataset (n=198) was used for model validation. Machine learning algorithms were applied to construct a diagnostic model. Performance was evaluated based on area under the curve (AUC) values, F1-score, negative predictive value, positive predictive value, accuracy, sensitivity, and specificity. RESULTS: The optimal model was based on a random forest algorithm with 10 clinical features. Thrombin time, prothrombin activity, and uric acid contributed most to the diagnostic model. The SLE diagnostic model showed sufficient predictive accuracy, with AUC values of 0.8286 in the validation dataset. CONCLUSIONS: Our diagnostic model based on 10 common laboratory tests identified the patients with SLE with high accuracy. An online version of the model can potentially be applied in clinical settings for the differential diagnosis of SLE.

Related factors of renal injury in primary Sjögren's syndrome.

BACKGROUND: Primary Sjögren's syndrome (pSS) is a common chronic systemic autoimmune disorder which primarily affects the exocrine glands. Patients may have extraglandular disease involving multiple organs, including the kidneys. This study aimed at investigating the clinical data and laboratory markers which were associated with renal function damage or renal involvement. METHOD: One thousand two hundred eighty-eight adult pSS patients from the Department of Rheumatology and Clinical Immunology were enrolled in this retrospective cohort study. And there were 334 patients of them followed up for more than two years for analyzing demographic, clinical data and laboratory markers. Statistical analysis was performed by R software (Version 3.6.2). RESULT: Nearly 95% of 1288 pSS patients were women, and the positive rates of anti-SSA (Sjögren's syndrome A) and anti-SSB were 63% and 27% respectively. 12% of the pSS patients presented renal involvement with eGFR < 60 mL/min/1.73 m2, and the mean age of hospital presentation, serum creatinine and urea were the highest (P < 0.001), and ANA (antinuclear antibody)-positive, anti-SSB-positive and anti-scl-70-positive were more prevalent in this group. Multivariate analyses showed that age, urea, chlorine and anti-SSA indicate a significant association with renal dysfunction. Potassium, sodium and Jo-1 were also confirmed to be related with decreased renal function. The receiver operating characteristic (ROC) analysis including the above factors showed a good performance on the evaluation of renal injury including eGFR < 60 mL/min/1.73 m2 and eGFR 60 -90 mL/min/1.73 m2 in pSS, with area under curve (AUC) values of 0.957 and 0.821, and high sensitivity (71.1% and 84.4%) and specificity (95.5% and 70.5%). After a more than two years follow-up of anti-SSA positive patients, 34.14% of them developed decreased renal function, and 13.58% of them experienced a progression of renal injury with a 23.64% decrease in eGFR. CONCLUSION: Age, urea, chlorine, and anti-SSA were highly associated with renal injury in pSS. Early screening for autoantibodies would be meaningful for evaluation and prevention of renal injury in pSS.

Study on phase evolution and promoting the pozzolanic activity of electrolytic manganese residue during calcination.

Electrolytic manganese residue (EMR) is a harmful by-product in the electrolytic manganese industry. Calcination is an efficient method for disposing EMR. In this study, thermogravimetric-mass spectrometry (TG-MS) combined with X-ray diffraction (XRD) was used for analysing the thermal reactions and phase transitions during calcination. The pozzolanic activity of calcined EMR was determined by the potential hydraulicity test and strength activity index (SAI) test. The leaching characteristics of Mn were determined by TCLP test and BCR SE method. The results showed that MnSO4 was converted into stable MnO2 during calcination. Meanwhile, Mn-rich bustamite (Ca0.228Mn0.772SiO3) was converted into Ca(Mn, Ca)Si2O6. The gypsum was transformed into anhydrite and then decomposed into CaO and SO2. Additionally, the organic pollutants and ammonia were completely removed following calcination at 700 °C. The leaching concentration of Mn decreased from 819.9 mg L-1 to 339.6 mg L-1 following calcination at 1100 °C. The chemical forms of Mn were transformed from acid-soluble fraction to residual fraction. The pozzolanic activity tests indicated that EMR1100-Gy maintained a complete shape. The compressive strength of EMR1100-PO reached 33.83 MPa. Finally, the leaching concentrations of heavy metals met the standard limits. This study provides a better understanding for the treatment and utilization of EMR.

Glutathione Synthesis Regulated by CtrA Protects Ehrlichia chaffeensis From Host Cell Oxidative Stress.

Ehrlichia chaffeensis, a small Gram-negative obligatory intracellular bacterium, infects human monocytes or macrophages, and causes human monocytic ehrlichiosis, one of the most prevalent, life-threatening emerging zoonoses. Reactive oxygen species are produced by the host immune cells in response to bacterial infections. The mechanisms exploited by E. chaffeensis to resist oxidative stress have not been comprehensively demonstrated. Here, we found that E. chaffeensis encodes two functional enzymes, GshA and GshB, to synthesize glutathione that confers E. chaffeensis the oxidative stress resistance, and that the expression of gshA and gshB is upregulated by CtrA, a global transcriptional regulator, upon oxidative stress. We found that in E. chaffeensis, the expression of gshA and gshB was upregulated upon oxidative stress using quantitative RT-PCR. Ehrlichia chaffeensis GshA or GshB restored the ability of Pseudomonas aeruginosa GshA or GshB mutant to cope with oxidative stress, respectively. Recombinant E. chaffeensis CtrA directly bound to the promoters of gshA and gshB, determined with electrophoretic mobility shift assay, and activated the expression of gshA and gshB determined with reporter assay. Peptide nucleic acid transfection of E. chaffeensis, which reduced the CtrA protein level, inhibited the oxidative stress-induced upregulation of gshA and gshB. Our findings provide insights into the function and regulation of the two enzymes critical for E. chaffeensis resistance to oxidative stress and may deepen our understanding of E. chaffeensis pathogenesis and adaptation in hosts.

Microbial Distribution and Antibiotic Susceptibility of Bloodstream Infections in Different Intensive Care Units.

Background: Bloodstream infection (BSI) is an increasing public health concern worldwide, representing a serious infection with significant morbidity and mortality, especially in children and the elderly. The predominant microbial distribution and antibiotic susceptibility were investigated among BSIs in the different intensive care units (ICUs)-pediatric ICU (PICU), surgical ICU (SICU), cardiac ICU (CICU), respiratory ICU (RICU), and geriatric ICU (GICU)-in order to achieve more efficient and appropriate therapies for patients in various ICUs. Methods: In this retrospective cross-sectional study, the blood specimens were collected from five different ICUs of Peking University First Hospital and comprehensive ICU of Miyun Teaching Hospital (Miyun ICU) before antimicrobial treatment from 2017 to 2020. Microorganism cultures of the blood samples were conducted, and positive cultures were tested for type of pathogens and drug susceptibility. Results: The prevalence of BSIs was the highest in the Miyun ICU (10.85%), followed by the RICU (9.48%) and the PICU (8.36%). The total prevalence of Gram-positive bacterial strains (especially Staphylococcus spp. and Enterococcus spp.) in the PICU (44.55%), SICU (57.58%), CICU (55.00%), GICU (49.06%), and Miyun ICU (57.58%) was higher than that of Gram-negative bacteria. The major bacterial strain was Acinetobacter baumannii in the PICU (21.82%); Klebsiella pneumoniae in the SICU (12.88%), CICU (30.00%), and RICU (30.39%); Escherichia coli in the GICU (20.75%); and Staphylococcus epidermidis (18.18%) in the Miyun ICU. Staphylococcus hominis of BSIs remained highly susceptible (>70%) to gentamicin, linezolid, daptomycin, teicoplanin, vancomycin, tigecycline, and rifampicin in all the ICUs. Its antibiotic sensitivity to levofloxacin was moderate in the PICU and CICU, but mild (<30%) in the SICU, RICU, and GICU. K. pneumoniae was highly susceptible to doxycycline, minocycline, and tigecycline in all the ICUs except the RICU, and its antibiotic sensitivity to imipenem, meropenem, amikacin, ciprofloxacin, and levofloxacin was high/moderate in the PICU, CICU, GICU, and Miyun ICU, but mild in the SICU and RICU. Conclusion: The current study demonstrated the distribution of prevalent microorganisms, and their antimicrobial susceptibility exhibited a high divergence among BSIs in different ICUs from a tertiary hospital and an outer suburban hospital in Beijing. Therefore, different antibiotic therapies for various wards and distinct age groups (especially between pediatric and elderly patients) should be considered to control the emergence and spread of highly antibiotic-resistant infections.

Sulodexide Prevents Peritoneal Fibrosis by Downregulating the Expression of TGF-ß1 and Its Signaling Pathway Molecules.

Peritoneal dialysis is one of the main renal replacement treatments. However, long-term peritoneal dialysis keeps the peritoneum in contact with the sugar-containing nonphysiological peritoneal fluid, which leads to recurrent peritonitis, peritoneal fibrosis, and failure of ultrafiltration. Transforming growth factor-ß1 (TGF-ß1), related cytokines, and inflammatory factors are closely related to peritoneal fibrosis. Sulodexide (SLX) is a new type of glycosaminoglycan preparation, which is involved in the formation of an anionic charge barrier and can maintain the selective permeability of vascular endothelial cells. In this study, the innovative analysis of SLX specifically prevents the process of peritoneal dialysis peritoneal fibrosis by downregulating the expression of TGF-ß1 and its signaling pathway molecules. We randomly divided 30 rats into three groups. The blank control group received no treatment. The peritoneal dialysis model group was injected with 4.25% peritoneal dialysate (PDF) 20 ml daily, and the SLX group was injected with 4.25% PDF 20 ml + sulodexide (SLX) 20 mg/kg daily. After 8 weeks of dialysis, the rats were sacrificed, and the peritoneal function test was performed to determine the amount of glucose transport and ultrafiltration. The thickness of peritoneal per unit area was observed under high magnification. The level of inflammation in peritoneal tissue and the expression of TGF-ß1/Smad were detected. The results showed that SLX can significantly improve peritoneal tissue thickening and inflammation, can downregulate the expression of TGF-ß1, Smad2, Smad3, and Smad7 in peritoneal tissue, and improve the progression of peritoneal fibrosis.

Immunological evaluation of an inactivated SARS-CoV-2 vaccine in rhesus macaques.

Because of the relatively limited understanding of coronavirus disease 2019 (COVID-19) pathogenesis, immunological analysis for vaccine development is needed. Mice and macaques were immunized with an inactivated severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine prepared by two inactivators. Various immunological indexes were tested, and viral challenges were performed on day 7 or 150 after booster immunization in monkeys. This inactivated SARS-CoV-2 vaccine was produced by sequential inactivation with formaldehyde followed by propiolactone. The various antibody responses and specific T cell responses to different viral antigens elicited in immunized animals were maintained for longer than 150 days. This comprehensive immune response could effectively protect vaccinated macaques by inhibiting viral replication in macaques and substantially alleviating immunopathological damage, and no clinical manifestation of immunopathogenicity was observed in immunized individuals during viral challenge. This candidate inactivated vaccine was identified as being effective against SARS-CoV-2 challenge in rhesus macaques.

Combination of Urine Exosomal mRNAs and lncRNAs as Novel Diagnostic Biomarkers for Bladder Cancer.

BACKGROUND: The recent discovery of miRNAs and lncRNAs in urine exosomes has emerged as promising diagnostic biomarkers for bladder cancer (BCa). However, mRNAs as the direct products of transcription has not been well evaluated in exosomes as biomarkers for BCa diagnosis. The purpose of this study was to identify tumor progression-related mRNAs and lncRNAs in urine exosomes that could be used for detection of BCa. METHODS: RNA-sequencing was performed to identify tumor progression-related biomarkers in three matched superficial tumor and deep infiltrating tumor regions of muscle-invasive bladder cancer (MIBC) specimens, differently expressed mRNAs and lncRNAs were validated in TCGA dataset (n = 391) in the discovery stage. Then candidate RNAs were chosen for evaluation in urine exosomes of a training cohort (10 BCa and 10 healthy controls) and a validation cohort (80 BCa and 80 healthy controls) using RT-qPCR. The diagnostic potential of the candidates were evaluated by receiver operating characteristic (ROC) curves. RESULTS: RNA sequencing revealed 8 mRNAs and 32 lncRNAs that were significantly upregulated in deep infiltrating tumor region. After validation in TCGA database, 10 markedly dysregulated RNAs were selected for further investigation in urine exosomes, of which five (mRNAs: KLHDC7B, CASP14, and PRSS1; lncRNAs: MIR205HG and GAS5) were verified to be significantly dysregulated. The combination of the five RNAs had the highest AUC to disguising the BCa (0.924, 95% CI, 0.875-0.974) or early stage BCa patients (0.910, 95% CI, 0.850 to 0.971) from HCs. The expression levels of these five RNAs were correlated with tumor stage, grade, and hematuria degrees. CONCLUSIONS: These findings highlight the potential of urine exosomal mRNAs and lncRNAs profiling in the early diagnosis and provide new insights into the molecular mechanisms involved in BCa.

Insights into the mechanism regulating the differential expression of the P28-OMP outer membrane proteins in obligatory intracellular pathogen Ehrlichia chaffeensis.

Ehrlichia chaffeensis causes human monocytic ehrlichiosis (HME), which is one of the most prevalent, life-threatening emerging infectious zoonoses. The life cycle of E. chaffeensis includes ticks and mammals, in which E. chaffeensis proteins are expressed differentially contributing to bacterial survival and infection. Among the E. chaffeensis P28-OMP outer membrane proteins, OMP-1B and P28 are predominantly expressed in tick cells and mammalian macrophages, respectively. The mechanisms regulating this differential expression have not been comprehensively studied. Here, we demonstrate that the transcriptional regulators EcxR and Tr1 regulate the differential expression of omp-1B and p28 in E. chaffeensis. Recombinant E. chaffeensis Tr1 bound to the promoters of omp-1B and p28, and transactivated omp-1B and p28 promoter-EGFP fusion constructs in Escherichia coli. The consensus sequence of Tr1 binding motifs was AC/TTATA as determined with DNase I footprint assay. Tr1 showed a higher affinity towards the p28 promoter than the omp-1B promoter as determined with surface plasmon resonance. EcxR activated the tr1 expression in response to a temperature decrease. At 37°C low level of Tr1 activated the p28 expression. At 25°C high level of Tr1 activated the omp-1B expression, while repressing the p28 expression by binding to an additional site upstream of the p28 gene. Our data provide insights into a novel mechanism mediated by Tr1 regulating E. chaffeensis differential gene expression, which may aid in the development of new therapeutics for HME.

Enhancing detection of steady-state visual evoked potentials using channel ensemble method.

Objective.This study proposed and evaluated a channel ensemble approach to enhance detection of steady-state visual evoked potentials (SSVEPs).Approach.Collected multi-channel electroencephalogram signals were classified into multiple groups of new analysis signals based on correlation analysis, and each group of analysis signals contained signals from a different number of electrode channels. These groups of analysis signals were used as the input of a training-free feature extraction model, and the obtained feature coefficients were converted into feature probability values using thesoftmaxfunction. The ensemble value of multiple sets of feature probability values was determined and used as the final discrimination coefficient.Main results.Compared with canonical correlation analysis, likelihood ratio test, and multivariate synchronization index analysis methods using a standard approach, the recognition accuracies of the methods using a channel ensemble approach were improved by 5.05%, 3.87%, and 3.42%, and the information transfer rates (ITRs) were improved by 6.00%, 4.61%, and 3.71%, respectively. The channel ensemble method also obtained better recognition results than the standard algorithm on the public dataset. This study validated the efficiency of the proposed method to enhance the detection of SSVEPs, demonstrating its potential use in practical brain-computer interface (BCI) systems.Significance. A SSVEP-based BCI system using a channel ensemble method could achieve high ITR, indicating great potential of this design for various applications with improved control and interaction.

Clinical manifestation and genetic findings in three boys with low molecular Weight Proteinuria - three case reports for exploring Dent Disease and Fanconi syndrome.

BACKGROUND: Dent disease is an X-linked form of progressive renal disease. This rare disorder was characterized by hypercalciuria, low molecular weight (LMW) proteinuria and proximal tubular dysfunction, caused by pathogenic variants in CLCN5 (Dent disease 1) or OCRL (Dent disease 2) genes. Fanconi syndrome is a consequence of decreased water and solute resorption in the proximal tubule of the kidney. Fanconi syndrome caused by proximal tubular dysfunction such as Dent disease might occur in early stage of the disease. CASE PRESENTATION: Three cases reported in this study were 3-, 10- and 14-year-old boys, and proteinuria was the first impression in all the cases. All the boys presented with LMW proteinuria and elevated urine albumin-to-creatinine ratio (ACR). Case 1 revealed a pathogenic variant in exon 11 of CLCN5 gene [NM_001127899; c.1444delG] and a nonsense mutation at nucleotide 1509 [p.L503*], and he was diagnosed as Dent disease 1. Case 2 carried a deletion of exon 3 and 4 of OCRL1 gene [NM_000276.4; c.120-238delG…A] and a nonsense mutation at nucleotide 171 in exon 5 [p.E57*], and this boy was diagnosed as Dent disease 2. Genetic analysis of Case 3 showed a missense mutation located in exon 2 of HNF4A gene [EF591040.1; c.253C > T; p.R85W] which is responsible for Fanconi syndrome. All of three pathogenic variants were not registered in GenBank. CONCLUSIONS: Urine protein electrophoresis should be performed for patients with proteinuria. When patients have LMW proteinuria and/or hypercalciuria, definite diagnosis and identification of Dent disease and Fanconi syndrome requires further genetic analyses.

Instance Transfer Subject-Dependent Strategy for Motor Imagery Signal Classification Using Deep Convolutional Neural Networks.

In the process of brain-computer interface (BCI), variations across sessions/subjects result in differences in the properties of potential of the brain. This issue may lead to variations in feature distribution of electroencephalogram (EEG) across subjects, which greatly reduces the generalization ability of a classifier. Although subject-dependent (SD) strategy provides a promising way to solve the problem of personalized classification, it cannot achieve expected performance due to the limitation of the amount of data especially for a deep neural network (DNN) classification model. Herein, we propose an instance transfer subject-independent (ITSD) framework combined with a convolutional neural network (CNN) to improve the classification accuracy of the model during motor imagery (MI) task. The proposed framework consists of the following steps. Firstly, an instance transfer learning based on the perceptive Hash algorithm is proposed to measure similarity of spectrogram EEG signals between different subjects. Then, we develop a CNN to decode these signals after instance transfer learning. Next, the performance of classifications by different training strategies (subject-independent- (SI-) CNN, SD-CNN, and ITSD-CNN) are compared. To verify the effectiveness of the algorithm, we evaluate it on the dataset of BCI competition IV-2b. Experiments show that the instance transfer learning can achieve positive instance transfer using a CNN classification model. Among the three different training strategies, the average classification accuracy of ITSD-CNN can achieve 94.7 ± 2.6 and obtain obvious improvement compared with a contrast model (p < 0.01). Compared with other methods proposed in previous research, the framework of ITSD-CNN outperforms the state-of-the-art classification methods with a mean kappa value of 0.664.

Data Augmentation for Motor Imagery Signal Classification Based on a Hybrid Neural Network.

As an important paradigm of spontaneous brain-computer interfaces (BCIs), motor imagery (MI) has been widely used in the fields of neurological rehabilitation and robot control. Recently, researchers have proposed various methods for feature extraction and classification based on MI signals. The decoding model based on deep neural networks (DNNs) has attracted significant attention in the field of MI signal processing. Due to the strict requirements for subjects and experimental environments, it is difficult to collect large-scale and high-quality electroencephalogram (EEG) data. However, the performance of a deep learning model depends directly on the size of the datasets. Therefore, the decoding of MI-EEG signals based on a DNN has proven highly challenging in practice. Based on this, we investigated the performance of different data augmentation (DA) methods for the classification of MI data using a DNN. First, we transformed the time series signals into spectrogram images using a short-time Fourier transform (STFT). Then, we evaluated and compared the performance of different DA methods for this spectrogram data. Next, we developed a convolutional neural network (CNN) to classify the MI signals and compared the classification performance of after DA. The Fréchet inception distance (FID) was used to evaluate the quality of the generated data (GD) and the classification accuracy, and mean kappa values were used to explore the best CNN-DA method. In addition, analysis of variance (ANOVA) and paired t-tests were used to assess the significance of the results. The results showed that the deep convolutional generative adversarial network (DCGAN) provided better augmentation performance than traditional DA methods: geometric transformation (GT), autoencoder (AE), and variational autoencoder (VAE) (p < 0.01). Public datasets of the BCI competition IV (datasets 1 and 2b) were used to verify the classification performance. Improvements in the classification accuracies of 17% and 21% (p < 0.01) were observed after DA for the two datasets. In addition, the hybrid network CNN-DCGAN outperformed the other classification methods, with average kappa values of 0.564 and 0.677 for the two datasets.

Microbial Distribution and Antibiotic Susceptibility of Lower Respiratory Tract Infections Patients From Pediatric Ward, Adult Respiratory Ward, and Respiratory Intensive Care Unit.

INTRODUCTION: Lower respiratory tract infections (LRTIs) account for significant morbidity and mortality in patients admitted to hospitals worldwide, especially in children and elderly. The prevalent microorganisms and antibiotic susceptibility were investigated among LRTI patients from the pediatric ward, adult respiratory ward, and respiratory intensive care unit (RICU) in order to achieve more efficient treatment protocols and better recovery. METHODS: In this retrospective cross-sectional study (January 2016 to December 2019), 4,161 positive culture samples out of 18,798 different specimens (9,645 respiratory tract samples and 9,153 blood samples) from LRTI patients were analyzed for pathogen incidence and antibiotic sensitivity. RESULTS: Among the respiratory tract cultures, the frequency of Gram-negative bacterial strains was higher than Gram-positive bacterial strains. Pseudomonas aeruginosa was the dominant pathogen in both the adult respiratory ward (n = 156, 21.49%) and RICU (n = 975, 35.67%), whereas Staphylococcus aureus (n = 66, 19.19%) was the most common bacterium in the pediatric ward. Among the blood cultures, Gram-positive bacteria remained the major microorganisms involved in LRTIs, and the most frequent pathogen was Staphylococcus epidermidis (n = 59, 47.20%) in the pediatric ward and Staphylococcus aureus (n = 10, 21.8%) in adult respiratory ward. However, Gram-negative bacteria were the main pathogens in the RICU, of which Klebsiella pneumoniae (n = 51, 27.57%) is the most prevalent. Pseudomonas aeruginosa of LRTI patients remained highly susceptible (>70%) to routine antibiotics in pediatric ward. However, it only had high susceptibility to amikacin, tobramycin, gentamicin in both the adult respiratory ward and RICU and its antibiotic sensitivity to meropenem and imipenem was moderate in the adult respiratory ward and mild (<30%) in the RICU. Staphylococcus aureus isolated from LRTI patients was highly susceptible to linezolid, daptomycin, teicoplanin, vancomycin, tigecycline, rifampicin, and trimethoprim/sulfamethoxazole in all three wards, moderately susceptible to gentamicin in both the adult respiratory ward and RICU and to clindamycin, oxacillin, moxifloxacin only in the adult respiratory ward. CONCLUSIONS: Microbial distribution and their patterns of antibiotic susceptibility revealed a high divergence among LRTI patients admitted to different wards in this hospital. Thus, different antibiotic therapies should be considered for distinct age groups.

Baseline Glycated Albumin Predicts the Renal Dysfunction in a Five-Year Prospective Population-Based Study.

BACKGROUND: Glycated albumin (GA) was reported to be associated with renal dysfunction in non-diabetic CKD population. This study assessed the correlation of GA and renal dysfunction and explored risk factors affecting renal progression in a general population-based study through a five-year follow-up. METHODS: Individuals who underwent a physical examination between September 2010 and September 2015 were enrolled. Multivariate linear regression was performed to assess the relationship between GA and eGFR change rate. The relationship between GA and renal progression was analyzed by multivariate logistic regression among 1,501 participants. Other risk factors were also explored and their predictive value was evaluated by ROC analysis, external validation was carried out in another 603 participants from the general population. RESULTS: The frequencies of subjects with renal progression increased obviously with the increment of baseline and mean GA according to quartile stratification (p for trend < 0.001). Baseline GA, age, and uric acid (p < 0.05) were identified as risk factors for renal dysfunction with a 30% or more decrease of eGFR. For every 1% increase of GA, the risk of deterioration of renal function increased to 1.585 in the population (95% CI, 1.299 - 1.935, p < 0.001). The predictive value of the model-building equation was confirmed by ROC analysis (AUC = 0.82, 95% CI: 0.773 - 0.832, p < 0.001) and in the validation group, predictive sensitivity and specificity were 85.7% and 73.5%. CONCLUSIONS: Baseline GA is independently associated with renal dysfunction. Uric acid and age are also considered risk factors. GA combining with age, serum creatinine and uric acid can serve as predictive indicators for the progression of renal dysfunction.

Crk1/2 and CrkL play critical roles in maintaining podocyte morphology and function.

Podocytes are actin-rich epithelial cells whose effacement and detachment are the main cause of glomerular disease. Crk family proteins: Crk1/2 and CrkL are reported to be important intracellular signaling proteins that are involved in many biological processes. However, the roles of them in maintaining podocyte morphology and function remain poorly understood. In this study, specific knocking down of Crk1/2 and CrkL in podocytes caused abnormal cell morphology, actin cytoskeleton rearrangement and dysfunction in cell adhesion, spreading, migration, and viability. The p130Cas, focal adhesion kinase, phosphatidylinositol 3-kinase/Akt, p38 and JNK signaling pathways involved in these alterations. Furthermore, knocking down CrkL alone conferred a more modest phenotype than did the Crk1/2 knockdown and the double knockdown. Kidney biopsy specimens from patients with focal segmental glomerulosclerosis and minimal change nephropathy showed downregulation of Crk1/2 and CrkL in glomeruli. In zebrafish embryos, Crk1/2 and CrkL knockdown compromised the morphology and caused abnormal glomerular development. Thus, our results suggest that Crk1/2 and CrkL expression are important in podocytes; loss of either will cause podocyte dysfunction, leading to foot process effacement and podocyte detachment.

Urine afamin and afamin-creatinine ratio as biomarkers for kidney injury.

AIM: The aim of this study was to evaluate the urine afamin (uAFM) and afamin-creatinine ratio (AfCR) levels in patients with glomerulonephritis. PATIENTS & METHODS: We determined uAFM and AfCR of 247 healthy volunteers and 129 biopsy-proven glomerulonephritis patients. RESULTS: Analytical evaluation study revealed the assay is a reliable and robust test for measuring uAFM. For reference intervals, uAFM and AfCR values were different significantly between males and females. uAFM and AfCR levels were significantly increased in patients with primary membranous nephropathy, IgA nephropathy and minimal change disease compared with healthy volunteers. uAFM and AfCR were positively correlated with urine albumin and albumin-creatinine ratio, respectively. CONCLUSION: Our study suggested that uAFM and AfCR may be attractive biomarkers for kidney injury.

Urine proteomics of primary membranous nephropathy using nanoscale liquid chromatography tandem mass spectrometry analysis.

BACKGROUND: Primary membranous nephropathy (PMN) is an important cause of nephrotic syndrome in adults. Urine proteome may provide important clues of pathophysiological mechanisms in PMN. In the current study, we analyzed and compared the proteome of urine from patients with PMN and normal controls. METHODS: We performed two technical replicates (TMT1 and TMT2) to analyze and compare the urine proteome from patients with PMN and normal controls by tandem mass tag (TMT) technology coupled with nanoscale liquid chromatography tandem mass spectrometry analysis (LC-MS/MS). Gene ontology (GO) enrichment analysis was performed to analyse general characterization of the proteins. The proteins were also matched against the database of Kyoto Encyclopedia of Genes and Genomes (KEGG). For validation, Western blot was used to analyze the selected proteins. RESULTS: A total of 509 proteins and 411 proteins were identified in TMT1 and TMT2, respectively. 249 proteins were both identified in two technical replicates. GO analysis and KEGG analysis revealed immunization and coagulation were predominantly involved. Among the differential protein, the overexcretion of alpha-1-antitrypsin (A1AT) and afamin (AFM) were validated by Western blot analysis. CONCLUSIONS: Our data showed the important role of immunologic mechanism in the development of PMN, and the value of urinary A1AT and AFM in biomarker discovery of patients with PMN. The discovery of the overexcretion of A1AT and AFM in the urine can help to further elucidate pathogenetic mechanisms involved in PMN.