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Epigenetics Dec 2024Lung cancer is one familiar cancer that threatens the lives of humans. circCTNNB1 has been disclosed to have regulatory functions in some diseases. However, the...
Lung cancer is one familiar cancer that threatens the lives of humans. circCTNNB1 has been disclosed to have regulatory functions in some diseases. However, the functions and related regulatory mechanisms of circCTNNB1 in lung cancer remain largely indistinct. The mRNA and protein expression levels were examined through real-time polymerase chain reaction (RT-qPCR) and western blot. The cell proliferation was tested through CCK-8 assay. The cell migration and invasion were confirmed through Transwell assays. The cell senescence was evaluated through SA-β-gal assay. The binding ability between miR-186-5p and circCTNNB1 (or YY1) was verified through luciferase reporter and RIP assays. In this study, the higher expression of circCTNNB1 was discovered in lung cancer tissues and cell lines and resulted in poor prognosis. In addition, circCTNNB1 facilitated lung cancer cell proliferation, migration, invasion, and suppressed cell senescence. Knockdown of circCTNNB1 retarded the Wnt pathway. Mechanism-related experiments revealed that circCTNNB1 combined with miR-186-5p to target YY1. Through rescue assays, YY1 overexpression could rescue decreased cell proliferation, migration, invasion, increased cell senescence, and retarded Wnt pathway mediated by circCTNNB1 suppression. Furthermore, YY1 acts as a transcription factor that can transcriptionally activate circCTNNB1 to form YY1/circCTNNB1/miR-186-5p/YY1 positive loop. Through in vivo assays, circCTNNB1 accelerated tumour growth in vivo. All findings revealed that a positive loop YY1/circCTNNB1/miR-186-5p/YY1 aggravated lung cancer progression by modulating the Wnt pathway.
Topics: YY1 Transcription Factor; Humans; MicroRNAs; Lung Neoplasms; Wnt Signaling Pathway; Cell Proliferation; Animals; Cell Line, Tumor; RNA, Circular; Cell Movement; Mice; Gene Expression Regulation, Neoplastic; Mice, Nude; Male; Disease Progression; Female; A549 Cells
PubMed: 38913848
DOI: 10.1080/15592294.2024.2369006 -
PloS One 2024Retinitis pigmentosa (RP) is the most common inherited retinal dystrophy and a major cause of blindness. RP is caused by several variants of multiple genes, and genetic...
Retinitis pigmentosa (RP) is the most common inherited retinal dystrophy and a major cause of blindness. RP is caused by several variants of multiple genes, and genetic diagnosis by identifying these variants is important for optimizing treatment and estimating patient prognosis. Next-generation sequencing (NGS), which is currently widely used for diagnosis, is considered useful but is known to have limitations in detecting copy number variations (CNVs). In this study, we re-evaluated CNVs in EYS, the main causative gene of RP, identified via NGS using multiplex ligation-dependent probe amplification (MLPA). CNVs were identified in NGS samples of eight patients. To identify potential CNVs, MLPA was also performed on samples from 42 patients who were undiagnosed by NGS but carried one of the five major pathogenic variants reported in Japanese EYS-RP cases. All suspected CNVs based on NGS data in the eight patients were confirmed via MLPA. CNVs were found in 2 of the 42 NGS-undiagnosed RP cases. Furthermore, results showed that 121 of the 661 patients with RP had EYS as the causative gene, and 8.3% (10/121 patients with EYS-RP) had CNVs. Although NGS using the CNV calling criteria utilized in this study failed to identify CNVs in two cases, no false-positive results were detected. Collectively, these findings suggest that NGS is useful for CNV detection during clinical diagnosis of RP.
Topics: Humans; Retinitis Pigmentosa; DNA Copy Number Variations; High-Throughput Nucleotide Sequencing; Female; Male; Eye Proteins; Middle Aged; Adult; Multiplex Polymerase Chain Reaction
PubMed: 38913662
DOI: 10.1371/journal.pone.0305812 -
Parasite (Paris, France) 2024Wild rodents are key carriers of various human pathogens, including Blastocystis spp. Our study aimed to assess the prevalence and genetic characteristics of...
Molecular investigation of Blastocystis sp. infections in wild rodents from the Inner Mongolian Autonomous Region and Liaoning province, China: High prevalence and dominance of ST4.
Wild rodents are key carriers of various human pathogens, including Blastocystis spp. Our study aimed to assess the prevalence and genetic characteristics of Blastocystis among wild rodents in the Inner Mongolian Autonomous Region and Liaoning Province of China. From November 2023 to February 2024, 486 rodents were captured in these regions. Fresh feces were collected from the intestines of each rodent for the isolation of DNA and PCR amplification of the vertebrate cytochrome b (cytb) gene to identify rodent species. Subsequently, PCR analysis and sequencing of the partial small subunit of the ribosomal RNA (rRNA) gene were utilized to detect Blastocystis in all fecal samples. Of the total samples, 27.4% (133/486) were found to be Blastocystis positive. The results revealed the presence of four species of rodents infected with Blastocystis, 32.3% (63/195) in Rattus norvegicus, 15.1% (16/106) in Mus musculus, 20.2% (18/89) in Apodemus agrarius, and 37.5% (36/96) in Cricetulus barabensis. Sequence analysis confirmed the existence of five Blastocystis subtypes: ST1 (n = 4), ST2 (n = 2), the ST4 (n = 125, the dominant subtype), ST10 (n = 1), and a novel ST (n = 1). The identified zoonotic subtypes (ST1, ST2, ST4, and ST10) highlight the possible role played by wild rodents in the transmission of Blastocystis to humans, thereby elevating the chances of human infection. Meanwhile, the discovery of novel sequences also provides new insights into the genetic diversity of this parasite.
Topics: Animals; Blastocystis; Blastocystis Infections; China; Prevalence; Feces; Rodent Diseases; Rats; Mice; Rodentia; Animals, Wild; Cytochromes b; Phylogeny; DNA, Protozoan; Polymerase Chain Reaction; Murinae
PubMed: 38912917
DOI: 10.1051/parasite/2024031 -
Biomolecules & Biomedicine Jun 2024Knee osteoarthritis (KOA) is one of the most common degenerative joint diseases in the elderly worldwide. The primary lesion in patients with KOA is the degeneration of...
Knee osteoarthritis (KOA) is one of the most common degenerative joint diseases in the elderly worldwide. The primary lesion in patients with KOA is the degeneration of articular cartilage. This study aimed to observe the biological effects of cyclic negative pressure on C28/I2 chondrocytes and to elucidate the underlying molecular mechanisms. We designed a bi-directional intelligent micro-pressure control device for cyclic negative pressure intervention on C28/I2 chondrocytes. Chondrocyte vitality and proliferation were assessed using Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) assays. The extracellular matrix was analyzed using real-time fluorescence quantitative polymerase chain reaction (PCR) and western blot, while the molecular mechanism of the chondrocyte response to cyclic negative pressure was explored through mRNA sequencing. Experimental data demonstrated that cyclic negative pressure promoted chondrocyte proliferation and upregulated the expression of chondrocyte-specific protein, namely the collagen type II alpha 1 chain (COL2A1) protein, and the transcription factor SRY-box transcription factor 9 (SOX9). Additionally, RNA sequencing analysis revealed that the gene levels of insulin-like growth factor 2 (IGF-2) and early growth response 1 (EGR-1) were significantly elevated in the cyclic negative pressure group. This study demonstrates that cyclic negative pressure stimulates the proliferation of C28/I2 chondrocytes by promoting the expression of EGR-1 and IGF-2. This new discovery may provide novel insights into cartilage health and KOA prevention.
PubMed: 38912889
DOI: 10.17305/bb.2024.10487 -
Journal of Applied Biomedicine Jun 2024In 2020, there were numerous cases in Kazakhstan with clinical symptoms of COVID-19 but negative PCR results in nasopharyngeal and oropharyngeal swabs. The diagnosis was...
In 2020, there were numerous cases in Kazakhstan with clinical symptoms of COVID-19 but negative PCR results in nasopharyngeal and oropharyngeal swabs. The diagnosis was confirmed clinically and by CT scans (computed tomography). The problem with such negative PCR results for SARS-CoV-2 infection confirmation still exists and indicates the need to confirm the diagnosis in the bronchoalveolar lavage in such cases. There is also a lack of information about confirmation of SARS-CoV-2 infection in deceased patients. In this study, various tissue materials, including lungs, bronchi, and trachea, were examined from eight patients who died, presumably from SARS-CoV-2 infection, between 2020 and 2022. Naso/oropharyngeal swabs taken from these patients in hospitals tested PCR negative for SARS-CoV-2. This study presents a modified RNA isolation method based on a comparison of the most used methods for RNA isolation in laboratories: QIAamp Viral RNA Mini Kit and TRIzol-based method. This modified nucleic acid extraction protocol can be used to confirm SARS-CoV-2 infection by RT-qPCR in the tissues of deceased patients in disputed cases. RT-qPCR with RNA of SARS-CoV-2 re-extracted with such method from post-mortem tissues that were stored at -80 °C for more than 32 months still demonstrated high-yielding positive results.
Topics: Humans; COVID-19; SARS-CoV-2; RNA, Viral; Male; Autopsy; Real-Time Polymerase Chain Reaction; Female; Lung; Middle Aged; Aged; COVID-19 Nucleic Acid Testing; Trachea; Adult; Nasopharynx
PubMed: 38912867
DOI: 10.32725/jab.2024.013 -
Heliyon Jun 2024Dental follicle cells (DFCs) promote bone regeneration and Circular RNAs (circRNAs) play crucial roles in bone development and regeneration. Our previous study...
Dental follicle cells (DFCs) promote bone regeneration and Circular RNAs (circRNAs) play crucial roles in bone development and regeneration. Our previous study demonstrated the upregulation of circFgfr2 expression during the osteogenic differentiation of DFCs. However, the molecular mechanisms and functional roles of circFgfr2 in DFCs osteogenesis remain unclear. In this study, we aimed to investigate the subcellular localization of circFgfr2 in DFCs using fluorescence hybridization. investigations demonstrated that circFgfr2 overexpression promoted osteogenic differentiation, as evidenced by real-time quantitative polymerase chain reaction. By integrating the outcomes of bioinformatics analyses, dual luciferase reporter experiments, and chromatin isolation by RNA purification, we identified circFgfr2 as a sponge for miR-133a-3p, a key regulator of osteogenic differentiation. Moreover, miR-133a-3p suppressed osteogenic differentiation by targeting DLX3 and RUNX2 in DFCs. We validated that circFgfr2 promoted the osteogenic differentiation of DFCs through the miR-133a-3p/DLX3 axis. To further investigate the therapeutic potential of circFgfr2 in bone regeneration, we conducted experiments and histological analyses. Overall, these results confirmed the crucial role of circFgfr2 in promoting osteogenesis. In summary, our findings demonstrated that the circFgfr2/miR-133a-3p/DLX3 pathway acts as a cascade, thereby identifying circFgfr2 as a promising molecular target for bone tissue engineering.
PubMed: 38912473
DOI: 10.1016/j.heliyon.2024.e32498 -
Frontiers in Microbiology 2024Clinical significance of coagulase-negative staphylococci (CoNS) has been gradually acknowledged in both healthcare and clinical research, but approaches for their...
INTRODUCTION
Clinical significance of coagulase-negative staphylococci (CoNS) has been gradually acknowledged in both healthcare and clinical research, but approaches for their precise discrimination at the species level remain scarce. The current study aimed to evaluate the association of CoNS with orthopedic infections, where accurate and prompt identification of etiology is crucial for appropriate diagnosis and treatment decision-making.
METHODS
A 16S rRNA-based quantitative PCR (qPCR) assay was developed for the detection of genus and two panels of 3-plex qPCR assays for further differentiation of six CoNS species with remarkable clinical significance, including , , , , , and . All the assays exhibited excellent analytical performance. ΔCq (quantification cycle) between 16S rRNA and CoNS species-specific targets was established to determine the primary CoNS. These methods were applied to detect CoNS in wound samples from orthopedic patients with and without infection.
RESULTS AND DISCUSSION
Overall, CoNS were detected in 17.8% (21/118) of patients with clinically suspected infection and in 9.8% (12/123) of patients without any infection symptom ( < 0.05). Moreover, the association with infection was found to be bacterial quantity dependent. was identified as the predominant species, followed by , , and . Male sex, open injury, trauma, and lower extremity were determined as risk factors for CoNS infections. CoNS-positive patients had significantly longer hospitalization duration (20 days (15, 33) versus 13 days (7, 22) for -negative patients, = 0.003), which could be a considerable burden for healthcare and individual patients. Considering the complex characteristics and devastating consequences of orthopedic infections, further expanding the detection scope for CoNS may be pursued to better understand the etiology of orthopedic infections and to improve therapeutic strategies.
PubMed: 38912353
DOI: 10.3389/fmicb.2024.1400096 -
Frontiers in Medicine 2024Tuberculosis is a contagious bacterial disease caused by . The emergence and spread of drug-resistant strains of in both developing and developed countries has made...
INTRODUCTION
Tuberculosis is a contagious bacterial disease caused by . The emergence and spread of drug-resistant strains of in both developing and developed countries has made diagnosis, treatment, and control of tuberculosis more difficult. The PCR assay, which is a fast and sensitive technique and an alternative method for detecting multidrug-resistant tuberculosis, is used to determine rifampicin (RIF) resistance. There is no single figure in Ethiopia that represents rifampicin-resistant tuberculosis and that is why this study was conducted to overcome the inconsistency of the results of the previous studies.
METHODS
Studies were researched from five major electronic databases. Studies which were cross-sectional in design, published, and written in English were included. The data were extracted using Microsoft Excel, and the data were managed and analyzed using Stata™ Version 17.0 statistical software. The Forest plot was used to check the presence of heterogeneity. The publication bias, meta-regression, and subgroup analysis were used to find out the source of heterogeneity. A random effect analysis model was used to pool the prevalence of RR TB from primary studies, and associated factors of RR among TB patients were identified using Meta regression. The presence of association was reported using OR with 95% CI.
RESULTS
The overall pooled prevalence of tuberculosis was 14.9% (95% CI: 13.34, 16.46), of these approximately 7.48% (95% CI: 6.30, 8.66) showed rifampicin-resistant tuberculosis in Ethiopia. Among the computed variables, 2.05% living with HIV1.39 (95%CI: 1.13, 1.72) and having a history of TB treatment (95%CI: 1.34, 3.15) were identified as significant factors associated with RR TB in Ethiopia.
CONCLUSION
Drug-resistant TB is one of the prevalent emerging infectious diseases among TB patients, which affects approximately one out of every thirteen TB patients. Having TB-HIV coinfection and a history of prior TB treatment were identified as significant factors associated with RR TB. To prevent and control RR TB, patients should complete their follow-up course; the health professionals should educate the actions taken by the patients when they experience drug toxicity and side effects; and the Minister of Health should initiate telemedicine and recruit tracers to overcome TB patients' default and have good drug adherence and retention after initiation of the treatment.
PubMed: 38912342
DOI: 10.3389/fmed.2024.1319845 -
International Journal of General... 2024The biological function and prognostic significance of endothelial cell specific molecule 1 (ESM1) in various cancers have been validated. This study aimed to explore...
BACKGROUND
The biological function and prognostic significance of endothelial cell specific molecule 1 (ESM1) in various cancers have been validated. This study aimed to explore the expression and clinical diagnosis values in patients with stomach adenocarcinoma (STAD) and esophageal carcinoma (ESCA).
METHODS
Online database Gene Expression Omnibus was used to screen for abnormally expressed genes in STAD and ESCA. Besides, 36 STAD and 36 ESCA patients were enrolled, and their corresponding control groups were also 36 people each. Reverse transcription-quantitative polymerase chain reaction and Western blot were performed to analyze the expression of ESM1. Overall survival (OS) curve and receiver operating characteristics curve (ROC) analysis were used to assess the prognosis, and the sensitivity and specificity of ESM1 for the diagnosis of STAD and ESCA, respectively. Additionally, the effects of ESM1 on cell viability, migration, and invasion were analyzed by cell counting kit-8, transwell migration and invasion assays.
RESULTS
The results showed that the poor OS of STAD and ESCA patients was correlated with high ESM1. Besides, ESM1 was increased in ESCA and STAD in in vivo and in vitro studies. ESM1 has a high accuracy [area under the curve (AUC) > 0.79] at stage I and IV of STAD and ESCA. Knockdown of ESM1 suppressed the cell viability, migration, and invasion and increased the apoptosis rate of AGS and TE1 cells.
CONCLUSION
Our study suggested that ESM1 might be used as a new indicator for the diagnosis and prognosis of early and advanced stage digestive tract cancers.
PubMed: 38912330
DOI: 10.2147/IJGM.S456973 -
Frontiers in Public Health 2024Upon the onset of the COVID-19 pandemic, the Public Health Laboratory Support Unit (ZIG4) at the Robert Koch Institute (RKI), the German National Public Health...
Capacity-building during public health emergencies: perceived usefulness and cost savings of an online training on SARS-CoV-2 real-time polymerase chain reaction (qPCR) diagnostics in low- and middle-income settings during the COVID-19 pandemic.
INTRODUCTION
Upon the onset of the COVID-19 pandemic, the Public Health Laboratory Support Unit (ZIG4) at the Robert Koch Institute (RKI), the German National Public Health Institute, developed and delivered an online training on SARS-CoV-2 qPCR diagnostics to 17 partner countries in low- and middle-income countries (LMIC). This article analyses the usefulness and cost savings of this training.
METHODS
The authors performed a concurrent mixed-methodology study based on key informant interviews, interviewer-administered questionnaires, and document reviews. Economic costs were estimated from the perspective of RKI.
RESULTS
Responding participants indicated that the course provided good and comprehensive information on up-to-date scientific knowledge and laboratory practice in PCR diagnostics. Respondents appreciated how the technical content of the training enhanced their ability to apply diagnostic methods in their daily work. Interviewees highlighted that the fast implementation and the low threshold of attending an online training had allowed them to quickly build skills that were crucial during, and beyond, the COVID-19 crisis. The total estimated cost of the online SARS-CoV-2 qPCR training was 61,644 euros. The total estimated cost of the equivalent face-to-face training was estimated at 267,592 euros. Programme weaknesses identified included the top-down approaches taken, lack of interactive components and opportunities to directly engage with other course participants and with teachers.
CONCLUSIONS
An online training was developed and implemented to support RKI partner countries in SARS-CoV-2 qPCR diagnostics during the COVID-19 pandemic, thereby strengthening pandemic response and health system resilience. The training incurred in important cost savings compared to the equivalent face-to-face training. Post-pandemic studies could usefully build on these research findings and explore ways to enhance end user involvement and improve interactive features to build stronger communities of learners and facilitate exchange of information and mutual learning.
Topics: Humans; COVID-19; SARS-CoV-2; Public Health; Cost Savings; Developing Countries; Capacity Building; Real-Time Polymerase Chain Reaction; Pandemics; Education, Distance; Surveys and Questionnaires; Germany
PubMed: 38912269
DOI: 10.3389/fpubh.2024.1197729