Clinical Samples Research Articles

Frequency of detection of Staphylococcus aureus isolates in various clinical materials from patients with purulent-inflammatory diseases in non-infectious clinics

Aim of the study. To study the frequency of isolation of Staphylococcus aureus from clinical biomaterials of patients with purulent-inflammatory diseases for the period from 2018 to 2022 according to the microbiological laboratory of JSC «National Scientific Medical Center», Astana.Materials and methods. A prospective microbiological study of the microbial landscape isolated from clinical samples of inpatients with purulent-inflammatory processes was carried out. Isolation and identification of strains was carried out on the basis of the microbiological laboratory of JSC «National Scientific Medical Center», Astana.Results and discussion. According to the microbiological laboratory of JSC «National Scientific Medical Center», during the period of bacteriological studies of clinical materials from patients with purulent-inflammatory diseases of various localization and from other materials used in medical institutions, from 2018 to 2022, a total of 961 strains of Staphylococcus aureus were isolated. When studying the dynamics of growth or decrease in the number over the years, it can be noted that for all the periods of study, we noted the most significant increase in the release of the amount of Staphylococcus aureus only from samples taken from the throat – 57.4%. In second place in terms of the frequency of Staphylococcus aureus discharge is a wound – 15%, in third place urine – 6.13%, followed by a nasal smear – 3.64% and sputum – 2.49%, respectively.Conclusion. Thus, one of the dominant pathogens in purulent-inflammatory diseases is Staphylococcus aureus and the main clinical biomaterial according to the frequency of detection during bacteriological examination of patients taken from non-infectious clinics from 2018 to 2022, according to the microbiological laboratory of JSC «NSMC» there was a throat smear, which accounted for 57.4%, then wound – 15% and urine – 6.13%. In the remaining biomaterials, the detection of Staphylococcus aureus was insignificant

Prevalence of Multidrug Resistance among Staphylococcus Species Isolated from Clinical Samples

Aims: One major thing that promotes antimicrobial resistance among bacteria is their potential to produce enzymes and biofilms, which remain noteworthy elements in their pathogenicity. This study aimed to determine the prevalence of multidrug resistance Staphylococcus species isolated from different clinical samples. Study Design: The study employed independent measures experimental study design. Place and Duration of Study: Staphylococcus species originating from clinical samples were obtained from the microbiology laboratory of two teaching hospitals in Ogbomosho, Oyo State, between April and October, 2023. Methodology: Twenty-six (26) pre-identified (by the hospital Laboratories) Staphylococcus spp. were obtained from the microbiology laboratories of two teaching hospitals in Ogbomoso, Nigeria. The isolates were subjected to microscopic, and biochemical tests to confirm their identities. The testing for antibacterial susceptibility was carried out using the Kirky-Bauer disk diffusion technique. A modified crystal violet biofilm assay was used to determined the ability of the isolates to produce biofilm. Molecular characterization was carried out to identify bacteria with very high resistance to the used antibiotics using 16S rRNA. The isolates were subjected to enzyme production assay like pectinase, protease, keratinase, cellulose and collagenase assay. Results: All the Staphylococcus species in this study showed varied degrees of prevalence. All the clinical bacterial isolates also showed 100% resistance to Amoxicillin/Clavulanate, Cefuroxime, Cloxicillin Meropenem, and Doxycycline, while the slightest resistance was observed for Gentamicin with 29.17%. The multiple antibiotic resistance index (MARI) for all the isolates was between 0.3 to 1.0, which is higher than the safe limit of 0.2, with a high percent (95.8%) of the bacteria being MAR. The result shows that 77.8% of these isolates could produce one or more enzymes. Conclusion: The biofilm and enzyme production abilities of the clinical bacteria were major factors that led to expanded resistance, as observed in this study.

Occurrence of Pathogenic and Allergenic Molds in the Outdoor and Indoor Environment of a Major Hospital and Molecular Epidemiology of Aspergillus fumigatus in Kuwait

Aspergilli and other molds are prevalent in the environment and are an important cause of opportunistic infections and seasonal allergies in susceptible patients. This study determined species distribution of various molds in outdoor/indoor air in and around a major hospital and performed antifungal susceptibility testing and molecular fingerprinting of environmental and clinical Aspergillus fumigatus isolates in Kuwait. Sampling for the isolation of molds was performed for a 17-month-period from the water/indoor air of medical/surgical wards/ICUs and outdoor air. Molds were identified by phenotypic characteristics and/or by the PCR-sequencing of rDNA/β-tubulin/calmodulin genes. Antifungal susceptibility testing was done by Etest. Fingerprinting was performed by nine-loci-based microsatellite analysis. A total of 6179 isolates were obtained from outdoor (n = 4406) and indoor (n = 1773) environments. These included Cladosporium spp. (n = 2311), Aspergillus spp. (n = 1327), Penicillium spp. (n = 1325), Paecilomyces spp. (n = 473), Alternaria spp. (n = 218), Bipolaris spp. (n = 133), and other molds (n = 392). Fingerprinting data revealed heterogeneity among clinical and environmental A. fumigatus and shared genotypes among outdoor air and hospital environmental isolates. Itraconazole-resistant A. fumigatus isolates with TR34/L98H mutations in Cyp51A were also recovered from outdoor air (n = 1), a hospital environment (n = 3), and clinical samples (n = 2). More than 15 fungal genera and all four Aspergillus (Nigri, Flavi, Fumigati, and Terrei) sections and nine rare aspergilli were detected. The isolation frequency was higher during the peak allergy season of October/November. The presence of shared genotypes among outdoor air and the hospital environment including triazole-resistant A. fumigatus suggests a reservoir for invasive infections among susceptible hospitalized patients.

CRISPR/Cas13a-Enhanced Porous Hydrogel Encapsulated Photonic Barcodes for Multiplexed Detection of Virus.

In this study, we present an ultrasensitive and specific multiplexed detection method for SARS-CoV-2 and influenza (Flu) utilizing CRISPR/Cas13a technology combined with a hydrogel-encapsulated photonic crystal (PhC) barcode integrated with hybridization chain reaction (HCR). The barcodes, characterized by core-shell structures, are fabricated through partial replication of periodically ordered hexagonally close-packed silicon dioxide beads. Consequently, the opal hydrogel shell of these barcodes features abundant interconnected pores that provide a substantial surface area for probe immobilization. Furthermore, the inherent structural colors remain stable during detection events due to the robust mechanical strength of the barcode cores. This integration of CRISPR/Cas13a and HCR leverages both the highly specific RNA recognition capabilities and trans-cleavage activity of Cas13a while employing HCR to enhance sensitivity. Upon encountering target RNA, Cas13a cleaves a hairpin probe, thereby initiating subsequent HCR amplification for enhanced detection sensitivity. Our method demonstrates high accuracy and sensitivity in multiplexed detection of SARS-CoV-2, Flu A and Flu B RNA with a limit-of-detection as low as 200 aM. Importantly, this assay also exhibits acceptable accuracy in repeated clinical sample testing. Thus, our platform represents a promising strategy for highly sensitive multiplexed virus detection in clinical.

Linezolid Dosing and Pharmacokinetics in North American Patients with Tuberculosis.

Linezolid is recommended in treatment regimens for rifampin- or multi-drug-resistant tuberculosis. However, considerable pharmacokinetic variability exists, and long-term use is limited by adverse effects. This study evaluates the pharmacokinetics of linezolid in patients with tuberculosis from an international therapeutic drug monitoring service. Linezolid trough, 2-hour, and 6-hour post-dose clinical samples from across North America were tested by the University of Florida Infectious Disease Pharmacokinetics Laboratory. Total serum concentrations were measured using liquid chromatography-tandem mass spectrometry. Therapeutic drug monitoring was performed, and measurements were compared to typical linezolid concentrations including a trough value of <2 mcg/mL and peak value between 12 and 26 mcg/mL. From January 2019 to December 2023, 1,604 linezolid samples from 500 patients and 817 unique TDM occasions were analyzed. Trough concentrations were measured on 670 samples (median 1.19 mcg/mL [range 0.00-20.06]), and 232 troughs (34.6%) were >2 mcg/mL. Among trough samples from linezolid dosing of 600mg daily or 5 days/weekly, 43.2% were >2 mcg/mL. Of 600 peak samples, 264 (44%) were outside the typical range, most (89%) being subtherapeutic at <12 mcg/mL. High serum linezolid trough concentrations were measured in approximately one-third of samples and in more than 40% of those taking the recommended dose. More than 40% of peak concentrations were outside typical range, 89% of which were <12 mcg/mL. This study demonstrates that therapeutic drug monitoring can be used to identify patients with serum linezolid concentrations outside of targeted ranges, allowing clinicians to make appropriate dose adjustments to improve outcomes.

TREM2 promotes the formation of a tumor-supportive microenvironment in hepatocellular carcinoma

BackgroundTriggering receptor expressed on myeloid cells 2 (TREM2), a surface receptor predominantly expressed on myeloid cells, is a major hub gene in pathology-induced immune signaling. However, its function in hepatocellular carcinoma (HCC) remains controversial. This study aimed to evaluate the role of TREM2 in the tumor microenvironment in the context of HCC progression.MethodsHCC was experimentally induced in wild-type (WT) and Trem2-deficient (Trem2−/−) mice, and clinical sample analysis and in vitro studies on macrophages were conducted. HCC cells were treated with conditioned medium from WT or Trem2−/− macrophages, and their malignant phenotypes and underlying mechanisms were analyzed.ResultsTREM2 deficiency reduced liver tumor burden in orthotopic and subcutaneous HCC models by altering CD8+ T cell infiltration. Trem2-deficient macrophages presented increased chemokine secretion. TGF-β1 was found to be positively correlated with TREM2 expression in HCC, and TGF-β blockade reversed TREM2 induction. On the other hand, TREM2+ macrophages were found to be associated with glycolysis and PKM2 expression in HCC cells; this association may be related to the secretion of IL-1β, which enhances the malignant phenotypes of HCC cells.ConclusionsThese results reveal that TREM2+ macrophages play a driving role in HCC progression by suppressing CD8+ T cell infiltration and promoting tumor cell glycolysis, providing a new therapeutic target for HCC.

Novel Isothermal Amplification Integrated with CRISPR/Cas13a and Its Applications for Ultrasensitive Detection of SARS-CoV-2.

We herein developed an ultrasensitive and rapid strategy to identify genomic nucleic acids by integrating a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 13a (Cas13a) into our recently developed isothermal technique, nicking and extension chain reaction system-based amplification (NESBA) reaction. In this technique, named CESBA, the NESBA reaction isothermally produces a large amount of RNA amplicons from the initial target genomic RNA (gRNA). The RNA amplicons bind to the crispr RNA (crRNA) and activate the collateral cleavage activity of Cas13a, which would then cleave the reporter probe nearby, consequently producing the final signals. Based on this design principle, we successfully detected SARS-CoV-2 gRNA as a model target very sensitively down to even a single copy (0.05 copies/μL) in both fluorescence- and lateral flow assay (LFA)-based modes with excellent specificity against other human coronaviruses (H-CoVs). We further validated the clinical applicability of CESBA by testing the 20 clinical samples with 100% clinical sensitivity and specificity. This work represents a potent and innovative strategy for the identification of genomic nucleic acids in molecular diagnostics, delivering exceptional levels of sensitivity.

Genetic profiling of extended-spectrum β-Lactamase and carbapenemase-producing Escherichia coli O157:H7 from clinical samples among diarrheal patients in Shashemene, Ethiopia

BackgroundEscherichia coli (E. coli) O157:H7, associated with diarrhea, poses a global health risk. In Ethiopia, where diarrhea is common, there is limited knowledge about these resistant strains and a lack of data on Extended-Spectrum β-Lactamase (ESBL) and carbapenemase production. Understanding the prevalence of antimicrobial resistance genes associated with ESBL and carbapenems is crucial for addressing diarrheal disease. This study aimed to investigate the genetic profile of ESBL and carbapenemase coding gene carriage in E. coli O157:H7 from clinical stool samples and evaluate antimicrobial susceptibility patterns.MethodsA total of twenty-nine bacterial isolates obtained from diarrheal patients were subjected to conventional culture and phenotypic (Kirby Bauer disc diffusion method) testing for antimicrobial resistance. Additionally, screening for the production of ESBL (combined disk method) and carbapenemase (modified carbapenem inactivation method) was conducted. Isolates that tested positive for ESBL and carbapenemase production were further analyzed, targeting five genes (blaNDM, blaKPC, blaCTX−M, blaTEM, and blaSHV) associated with ESBL and carbapenemase production. Data analysis was performed using SPSS version 27.0, employing logistic regression and descriptive statistics.ResultsWe analyzed a total of 27 isolates that were ESBL-positive and 12 isolates that were found to produce carbapenemase phenotypically. These isolates were obtained from clinical stool samples and (9/27) 33.3% of the isolates were from under five years children, predominantly from urban areas, and those that have contact with domestic animals. Genes coding ESBL were found in (19/27) 70.4% of the isolates, the most predominant being blaCTX−M and blaTEM. Eight isolates carried blaKPC, but none had blaNDM, while five isolates carried both blaCTX−M and blaTEM genes. blaSHV-carrying isolates showed phenotypic resistance to ampicillin and cephalosporins, while blaKPC-carrying isolates exhibited resistance to ampicillin, carbapenems, and tetracycline.ConclusionThis study identifies a significant prevalence of multidrug resistance in E. coli O157:H7, which can be attributed to the presence of resistance genes coding for ESBL and carbapenem production. Key factors contributing to this resistance, such as urban environments, children under the age of five, and domestic animal ownership, have been emphasized. Additionally, this research underscores the urgent need for enhanced surveillance and targeted interventions to address this pressing public health concern.

A TaqMan-MGB Probe Quantitative PCR Assay Detecting Hematodinium perezi.

Hematodinium perezi, a pathogenic dinoflagellate, is one of major epidemiological agents that lead to severe losses of cultured marine crustaceans in China. This study aimed to develop a novel, sensitive and specific detection method qualified for early surveillance and control of the disease caused by H. perezi. The present study established a TaqMan-MGB probe quantitative PCR (qPCR) method, targeting the first internal transcribed spacer 1 (ITS 1) region of H. perezi by optimising reaction components. A high correlation coefficient (R2 = 0.9996) was obtained in a standard curve with a 103.4% efficiency. No amplification was observed for the host's genome and pathogens other than H. perezi in the TaqMan-MGB probe qPCR assays, showing high specificity to H. perezi. When using the plasmid standard DNA as templates, the detection limit of the qPCR method was determined to be 5.66 copies/reaction and 10 times more sensitive than the conventional PCR. The TaqMan-MGB probe qPCR assays exhibited high repeatability, and the intra- and inter-assay coefficients of variation (CV) ranged from 0.11% to 2.25% over a wide dynamic range of detection from 5.66 × 100 to 5.66 × 109 copies of targeting gene. The application was also validated on clinical samples, including those with low infection with H. perezi. This novel one-step TaqMan-MGB probe qPCR provides an option for surveillance and epidemiological investigations of H. perezi infection, with an advantage at the early infection stage.

Lipid Metabolism Related Gene ACSL3 as a Biomarker for Predicting Immunotherapy Outcomes in Lung Adenocarcinoma.

Investigate the role of lipid metabolism in the tumor immune microenvironment (TIME) of lung adenocarcinoma (LUAD) and identify vital lipid metabolism-related genes (LMRGs) that contribute to immunotherapy outcomes. 1130 LUAD patients were acquired utilizing public databases. Multiple algorithms were used to analyze the contribution of lipid metabolism in TIME. Importantly, cell lines, clinical samples (52 patients in surgery cohort and 36 in immunotherapy cohort), animal models, RNA-seq, experiments in protein and mRNA levels were conducted for identifying and validating key biomarker in LUAD immunotherapy. A prognostic signature comprising 33 LMRGs was developed and validated as an effective predictor of prognosis and TIME, with a C-index of 0.766 (95% CI: 0.729-0.804). Additionally, we identified Acyl-CoA Synthetase Long Chain Family Member 3 (ACSL3) as a potential biomarker for immunotherapy prognosis. The expression of ACSL3 was verified in 88 clinical tissues from LUAD patients, which indicated that elevated ACSL3 expression was correlated with worse progression-free survival (PFS) (p<0.001) and overall survival (OS) (p=0.008). Subsequent experiments revealed that knockdown of ACSL3 in vivo enhanced the efficacy of immunotherapy, potentially through increasing interferon α secretion, as indicated by Bulk RNA-seq and ELISA analysis, thereby promoting the infiltration of anti-tumor immune cells. The study established a model that accurately predicts immunotherapy response, prognosis, and TIME dynamics in LUAD patients. Notably, the pivotal role of ACSL3 in driving tumor progression and immune evasion was uncovered, offering novel insights into the optimization of immunotherapy strategies for LUAD.

ScRICA: An R package for multiple-sample single-cell RNA-seq data integrative comparative analysis

Single-cell sequencing technologies offer unprecedented resolution to inspect transcriptomes and generate critical biological insights. As the number of cells and cell types increase in single-cell studies, the effort required to analyze the data surges dramatically, especially when comparative explorations need to be performed on large datasets with different cell types and various sample attributes, such as clinical samples from different age and ancestry groups. Due to the sequential nature of single-cell data analysis, many steps involving multiple method choices and parameter options need to be considered. The computational skills required for integrative and comparative analyses of large datasets with various sample attributes represent a substantial obstacle for many researchers. To address this challenge, we have developed scRICA, a systematic workflow tailored for integrative and comparative single-cell RNA sequencing (scRNA-seq) analysis. This approach streamlines the analytical process, ensuring efficient utilization of computational resources and facilitating scalability for large-scale datasets. With scRICA, researchers can conduct integrative and comparative scRNA-seq analyses with ease, empowering them to derive meaningful insights from their data in a timely manner. scRICA offers a versatile approach by allowing users to input various parameter options from a metadata table, which are inherited throughout the entire analysis workflow. This functionality greatly enhances the efficiency of programming for comparative analyses involving multiple sample attributes. As an R package, scRICA provides a user-friendly interface within the R environment, making it accessible to researchers familiar with R programming. Additionally, scRICA offers a command line execution option, allowing users to seamlessly integrate it into their computational pipelines or execute analyses on High-Performance Computing (HPC) systems. This combination of features ensures flexibility, ease of use, and scalability, making scRICA a valuable tool for comprehensive and efficient single-cell RNA sequencing analysis.

Nonylphenol exposure increases the risk of Hirschsprung's disease by inducing macrophage M1 polarization.

Nonylphenol (NP), a ubiquitous environmental contaminant used as a surfactant in industrial production and classified as an endocrine disruptor, could interfere hormone secretion and exhibit neurotoxicity in organisms. Hirschsprung's disease (HSCR), one of the most frequently observed congenital malformations of the digestive system, arises mainly due to the failure of enteric neural crest cells to migrate to the distal colon during embryonic development. However, the effects of NP exposure on HSCR are largely unknown. Herein, we identified the content of NP and expression of lncRNA LINC00294/Inhibin Subunit Beta E (INHBE) axis in clinical samples and evaluated the crucial role of lncRNA LINC00294/INHBE axis in the neurogenic potential of neurons and the neurotoxicity effects of NP in the SH-SY5Y cells and female specific pathogen-free (SPF) rat model. Our results showed that NP concentration and LINC00294 were significantly associated with HSCR occurrence and macrophage polarization in human HSCR specimens. Moreover, NP promoted macrophage M1 polarization. The proliferation and migration were weakened, and apoptosis was heightened by the conditioned medium of NP-treated macrophages in SH-SY5Y cells. Contrarily, LINC00294 overexpression and INHBE knockdown reversed the neurotoxicity effect of NP on SH-SY5Y cells. Furthermore, the neurotoxicity effect of NP was abolished by clodronate liposomes in the rat model. In conclusion, NP could induce macrophage M1 polarization via the LINC00294/INHBE axis and increase the risk of Hirschsprung's disease. Our findings would provide a foundation for the toxicity study and risk assessments of NP.

Posttraumatic Stress Disorder and Dissociation in a Clinical Sample of Refugees in the Netherlands: Evidence for a Dissociative Subtype

ABSTRACT An increasing number of studies have been investigating the co-occurrence of posttraumatic symptoms and dissociation in trauma-exposed samples. As traumatized refugees are particularly susceptible to developing posttraumatic stress disorder (PTSD), the aim of this study was to investigate the relationship between PTSD and dissociation in a traumatized refugee sample. Cross-sectional data from a clinical refugee sample (N = 526) were collected. Latent class analysis (LCA) examined different classes of PTSD, based on the Clinician Administered PTSD Scale for DSM-5 (CAPS-5) items. Subsequently, it was examined whether cumulative trauma, sexual trauma and general psychopathology predicted class membership. The LCA identified five classes. The classes were summarized as (1) “High PTSD,” (2) “Moderate PTSD,” (3) “High PTSD with high loss of interest,” (4) “High PTSD with moderate loss of interest,” and (5) “PTSD-DS.” PTSD DS (10% of the sample) was characterized by high PTSD symptoms, as well as high depersonalization and derealization symptoms. The majority (61.4%) of this group has been exposed to sexual trauma. Overall endorsement of PTSD symptoms was extremely high in this clinical sample of refugees. A group evidencing the PTSD dissociative subtype was identified.

EFHD1 Activates SIK3 to Limit Colorectal Cancer Initiation and Progression via the Hippo Pathway.

Colorectal cancer (CRC) is one of the most commonly diagnosed cancers, with high rates of metastasis and lethality. EF-hand domain-containing protein D1 (EFHD1) and salt-inducible kinase 3 (SIK3) have been studied in several cancer types. Aberrant expression of EFHD1 and SIK3 has been observed in CRC, but little research has addressed their regulatory abilities and signaling pathways. In this study, we aimed to explore the efficacy of EFHD1 in inhibiting CRC proliferation and metastasis and to elucidate the underlying mechanisms involved in the upregulation of SIK3 expression. Cell viability, colony formation, wound healing, Transwell assay, orthotopic xenograft, and pulmonary metastasis mouse models were used to detect the antiproliferative and anti-metastatic effects of EFHD1 against CRC in vitro and in vivo. The Gene Expression Profiling Interactive Analysis (GEPIA) database was used to determine EFHD1 and SIK3 expression in CRC. The regulatory roles of EFHD1 and SIK3 in mediating anti-metastatic effects in CRC were measured using western blotting, immunohistochemical, and immunofluorescence analyses. The results showed that EFHD1 expression was significantly repressed in the clinical CRC samples. EFHD1 markedly suppressed cell proliferation, migration, and invasion in vitro and inhibited tumor growth and metastasis in vivo. Analysis of the GEPIA database revealed that EFHD1 expression positively correlated with SIK3 expression. SIK3 overexpression inhibited the migration of CRC cells, and SIK3 knockdown partially eliminated the inhibitory effects of EFHD1 on CRC metastasis. EFHD1 exerted anti-metastatic effects against CRC via upregulating SIK3 and inhibiting epithelial-mesenchymal transition (EMT) processing through modulating the Hippo signaling pathway. Collectively, these findings identify EFHD1 as a potent SIK3 agonist and highlight the EFHD1-SIK3 axis as a key modulator of the Hippo signaling pathway in CRC. EFHD1 serves as a novel regulator and is worthy of further development as a novel therapeutic target in CRC.

A green impedimetric aptasensor for non-invasive and high-selective detection of tramadol validated by molecular dynamic simulation.

Although the dosage controlling of tramadol (TRA) as a banned deadly drug in human biofluids is medicolegally important a biocompatible method for its high-selective detection with fewer false interferences has been scarcely reported. Herein, a new impedimetric aptasensor is introduced by utilizing the aptamer (Apt) sequence with high affinity to TRA for the first time to non-invasively measure it. An oriented nanolayer of Au nanoparticles (AuNPs) is easily formed on the surface by the electrodeposition technique to high-densely load the Apt and embed the novel aptasensing interface via a user-friendly methodology. The visual interaction of Apt with its target has been explored using molecular dynamic (MD) simulation to confirm how Apt traps TRA in its arm. The aptasensor measured TRA in a concentration range of 50 pM to 1.3nM with a limit of detection (LOD) of 16.66 pM in buffer. It also rendered good accuracy and recovery for human salivary and urinary analysis. In addition, the greenness profile of the proposed methodology has been validated with two international common indexes. The developed aptasensor promises a reasonable capability for TRA monitoring in real clinical or street narcotic samples according to green analytical chemistry (GAC).

MOF scaffold for anchoring platinum-nickel nanoparticles with enhanced oxidase-like activity to improve lateral flow immunoassay diagnosis.

Noble metal nanoparticles have attracted tremendous attention as the promising signal reporters for catalytic-colorimetric lateral flow immunoassay (LFIA). However, it remains great challenges for improving their stability and catalytic activity. Herein, first, a kind of porphyrinic based metal-organic framework (MOF) was used as a carrier for loading platinum (Pt) nanoparticles to avoid its aggregation. Moreover, nickel (Ni) atoms were dopped into Pt nanoparticles to adjust crystal structure, thus greatly improving catalytic activity. The resulting MOF@PtNi nanocomposite showed enhanced colorimetric signal brightness and excellent oxidase-like activity, which can improve sensitivity via amplifying the color signal. The catalytic mechanism was further studied by scavenger and electron paramagnetic resonance analysis. Furthermore, integrated with the competitive immunization LFIA platform, the high sensitivity colorimetric detection of human immunoglobulin G was realized with a detection limit of 0.378ng/mL and 0.269ng/mL for pre- and post-catalytic detection, respectively. In addition, this MOF@PtNi based catalytic-colorimetric LFIA was used for detection of clinical serum samples and the results agreed well with that measured by the standard method. Therefore, this study helps open up the application of proposed catalytic-colorimetric nanocomposite in the ultrasensitive LFIA for point-of-care diseases diagnosis.

CXCL12 is a potential therapeutic target for type 2 diabetes mellitus complicated by chronic obstructive pulmonary disease.

To identify the key genes and immunological pathways shared by type 2 diabetes mellitus (T2DM) and chronic obstructive pulmonary disease (COPD) and explore the potential therapeutic targets of T2DM complicated by COPD. GEO database was used for analyzing the gene expression profiles in T2DM and COPD to identify the common differentially expressed genes (DEGs) in the two diseases. A protein-protein interaction network was constructed to identify the candidate hub genes, which were validated in datasets and disease sets to obtain the target genes. The diagnostic accuracy of these target genes was assessed with ROC analysis, and their expression levels and association with pulmonary functions were investigated using clinical data and blood samples of patients with T2DM and COPD. The abundance of 22 immune cells was analyzed with CIBERSORT algorithm, and their relationship with the target genes was examined using correlation analysis. DGIdb database was used for analyzing the drug-gene interactions and the druggable genes followed by gene set enrichment analysis. We identified a total of 175 common DEGs in T2DM and COPD, mainly enriched in immune- and inflammation-related pathways. Among these genes, CXCL12 was identified as the final target gene, whose expression was elevated in both T2DM and COPD (P<0.05) and showed good diagnostic efficacy. Immune cell infiltration correlation analysis showed significant correlations of CXCL12 with various immune cells (P<0.01). GESA analysis showed that high CXCL12 expression was significantly correlated with "cytokine-cytokine receptor interaction". Drug-gene analysis showed that most of CXCL12-related drugs were not targeted drugs with significant cytotoxicity. CXCL12 is a potential common key pathogenic gene of COPD and T2DM, and small-molecule targeted drugs against CXCL12 can provide a new strategy for treatment T2DM complicated by COPD.

Development of a Portable Rapid Detection Method for Porcine Epidemic Diarrhea Virus Using Reverse Transcription-Recombinase-Aided Amplification Technology.

Porcine epidemic diarrhea virus (PEDV) continues to spread globally, causing clinical symptoms in piglets, including watery diarrhea, vomiting, and dehydration. Its exceptionally high morbidity and mortality rate contributes significantly to the economic losses of the swine industry. The continuous genetic mutations of PEDV have compromised the effectiveness of classical strain vaccines. Early detection and accurate diagnosis are therefore crucial for controlling its further spread. Developing a detection method that is user-friendly, highly sensitive, and efficient is crucial for disease control. In this study, a point-of-care rapid detection method for PEDV was successfully established using reverse transcription-recombinase-aided amplification (RT-RAA) technology. This method enables results to be obtained within 20 min of amplification at a constant temperature of 42 °C. It demonstrates high sensitivity, with a detection limit as low as 1 copy/μL, and shows strong specificity, with no cross-reactivity observed with seven other common swine pathogens. When applied to clinical samples, the results were 100% consistent with those obtained by RT-qPCR. This method is distinguished by its portable instrumentation and simple operation, making it particularly suitable for resource-constrained settings.

A nucleocapsid monoclonal antibody based sandwich ELISA for the general detection of both PRRSV-2 and PRRSV-1.

Porcine reproductive and respiratory syndrome virus (PRRSV) causes reproductive failure in sows and respiratory disease in growing pigs, leading to significant economic losses worldwide. Due to the constant mutation and recombination, PRRSV exhibits significant genetic diversity, the general detection of all PRRSV-2 and PRRSV-1 strains is thus needed. In our study, four monoclonal antibodies (mAbs) against PRRSV nucleocapsid (N) protein were generated and the precise and novel B cell epitopes (52KPHF55 and 109HHTVR113) were identified. The epitope 52KPHF55 is highly conserved across all strains of PRRSV-2 lineages and PRRSV-1 subtypes, and the corresponding two mAbs (6D7, 4D12) were selected to develop a sandwich ELISA that was able to detect all tested PRRSV-2 and PRRSV-1 strains. The developed sandwich ELISA demonstrated high specificity, sensitivity and repeatability. In detection of 133 clinical samples, the sandwich ELISA achieved 84.21 % coincidence with the real-time RT-PCR. In conclusion, the mAb based sandwich ELISA can be suitable for detection of potential all PRRSV-2 lineages and PRRSV-1 subtypes, providing a simple, quick and high content method for diagnosis of PRRS.

Overexpression of CHMP2B suppresses proliferation of renal clear cell carcinoma cells.

To analyze the association of CHMP2B expression level of with clinicopathological characteristics and prognosis of clear cell renal cell carcinoma (CRCC) and the possible role of CHMP2B in tumorigenesis and progression of CRCC. RNAseq data of CRCC were downloaded from the TCGA database for analysis of CHMP2B expression levels in tumor and adjacent tissues and their correlation with clinicopathological characteristics of the patients. Survival outcomes of the patients with high and low CHMP2B expressions were analyzed using the Kaplan-Meier model, and the COX risk regression model was used for identifying the prognostic factors of the patients. The correlation between CHMP2B expression and immune infiltration, its co-expressed genes, and the effect of CHMP2B gene mutations on immunotherapy responses, and its immunohistochemical expression in CRCC and normal tissues were analyzed. Clinical samples of CRCC were collected to examine CHMP2B expressions using RT-PCR, and cell experiment was carried out to test the effect of CHMP2B overexpression on biological behaviors of CRCC cells. CHMP2B was significantly under-expressed in renal cancer tissues, and its overexpression obviously inhibited the proliferation of CRCC cells in vitro. CHMP2B expression level was significantly correlated with age, gender, lymph node metastasis, and tumor stage, and the patients with low CHMP2B expression had poor survival outcomes. Enrichment and co-expression gene analyses suggested that CHMP2B was mainly involved in viral outgrowth, necrotic apoptosis, endocytosis, and immune-regulatory processes in kidney cancer. CHMP2B is lowly expressed in renal cancer tissues to affect tumor progression and tumor immune processes, and may serve as a prognostic biomarker and therapeutic target for CRCC.