Open Life Sciences最新文献

A growing body of evidence has shown that hepatitis B surface antigen (HBsAg) mutations can influence the occurrence of occult hepatitis B infection (OBI), particularly amino acid substitutions in small hepatitis B surface proteins (SHBs). The mechanistic basis for these results, however, remains unclear. This study was designed to explore the potential impact and mechanisms of OBI-related SHBs mutations on serum HBsAg. Huh7 and HepG2 cells were transfected with plasmids encoding wild-type (WT) or OBI-related SHB mutation-containing sequences, after which a chemiluminescence approach was used to detect HBsAg levels in cell culture supernatants. Western blotting was further used to assess HBsAg and endoplasmic reticulum stress (ERS)-related protein levels in lysates prepared from these cells, while the localization of HBsAg within cells was assessed via immunofluorescent staining. Cells transfected with OBI-related SHB mutation-encoding plasmids exhibited lower supernatant HBsAg levels than cells transfected with WT plasmids. Intracellular and extracellular HBsAg levels in these mutant plasmid-transfected cells were lower relative to those for WT plasmid-transfected cells, and HBsAg accumulation within the ER was detected via immunofluorescent staining in cells transfected with OBI-related SHB mutation-encoding plasmids, ERS-related protein content was also significantly increased in mutant plasmid-transfected cells as compared to those in the WT group. These results suggest that proteins harboring OBI-related mutations may tend to accumulate in the ER, thereby triggering an ERS response and impairing the transcription and translation of HBsAg via the activation of the unfolded protein response and ER-associated protein degradation pathway. These effects ultimately reduce the overall assembly of HBV virions in the ER and their associated secretion.

Genomic DNA (gDNA) extraction is an important step in many molecular studies of fungal biology, and it is necessary to evaluate the efficiency, cost-effectiveness, and efficacy of different extraction methods to ensure successful amplification of the target gene and minimize deoxyribonucleic acid (DNA) degradation. The modified cetyltrimethylammonium bromide (CTAB) method was found to be effective in releasing high molecular weight gDNA with minimal protein contamination. Based on anticipated gDNA yield and quality, extraction time, cost effectiveness, successful amplification, and waste management, our findings serve as a guide for selecting techniques of gDNA extraction from fungal species. This study presents a modified CTAB method for extracting DNA from a variety of fungal species including Aspergillus, Penicillium, Alternaria, Dothiorella, and Fusarium. Comparison of three cell crushing methods reveals similar gDNA yields, demonstrating the method's effectiveness. Furthermore, the modified CTAB method is cost-effective and safe, eliminating the need for grinding with liquid nitrogen or bead beating. The method has a potential use for nucleic-based fungal disease diagnosis such as fish fungal diseases, plant pathogens, fruit rot associated pathogens, and human fungal diseases as we were successful in PCR amplifying several gene loci from varied fungal pathogens.

Dental pulp stem cells hold significant prospects for tooth regeneration and repair. However, a comprehensive understanding of the molecular differences between dental pulp stem cells (DPSC, from permanent teeth) and stem cells from human exfoliated deciduous teeth (SHED, from deciduous teeth) remains elusive, which is crucial for optimizing their therapeutic potential. To address this gap, we employed a novel data-independent acquisition (DIA) proteomics approach to compare the protein expression profiles of DPSC and SHED. Based on nano-LC-MS/MS DIA proteomics, we identified over 7,000 proteins in both cell types. By comparing their expression levels, 209 differentially expressed proteins were identified. Subsequent Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses, along with protein-protein interaction network construction, revealed significant metabolic differences and key regulatory nodes. DPSC exhibited significantly higher expression of proteins belonging to the NDUFB family, SMARC family, RPTOR and TLR3. These proteins are known to be involved in critical cellular processes such as mitochondrial energy metabolism, mTOR-related autophagy pathway, and innate immune response. Conversely, SHED displayed elevated expression of AKR1B family, which participated in glycerolipid metabolism and adipogenic differentiation, PRKG1, MGLL and UQCRB proteins associated with thermogenesis. These findings highlight the specific proteomic landscape of DPSC and SHED, suggesting their distinct biological roles and potential applications.

Neonatal sepsis (NS) is highly likely to cause death; however, early diagnosis of NS is still a great challenge. This study aimed to determine the diagnostic values of IL-6, IL-8, and serum amyloid A (SAA) in NS patients. C-Reactive protein (CRP), procalcitonin (PCT), interleukin (IL)-6, IL-8, and SAA were detected in 120 infants with NS (60 premature infants [NS-PIs] and 60 term infants [NS-TIs]). Sixty noninfected premature infants and 60 noninfected term infants composed the control group. Receiver operating characteristic (ROC) curves were used to determine the sensitivity and specificity of IL-6, IL-8, and SAA alone and in combination with CRP and PCT. The area under the curve (AUC) was calculated to evaluate the diagnostic value. The clinical characteristics of the subjects were recorded. The expression of CRP, PCT, IL-6, IL-8, and SAA was upregulated in patients with NS compared with control subjects. When the SAA cut-off value was 10.18 mg/L, the greatest AUC for the diagnosis of NS-PIs was for SAA (AUC = 0.833, 95% CI 0.762-0.905, P < 0.001). When the CRP cut-off value was 9.562 mg/L, the smallest AUC for the diagnosis of NS-PIs was for CRP (AUC = 0.776, 95% CI 0.684-0.867, P < 0.001). When the IL-8 cut-off value was 52.03 pg/mL, the greatest AUC for the diagnosis of NS-TIs was for IL-8 (0.821). When the IL-8 cut-off value was 52.03 pg/mL, the greatest AUC for the diagnosis of NS-TIs was for IL-8 (AUC = 0.821, 95% CI 0.745-0.898, P < 0.001). When the CRP cut-off value was 13.18 mg/L, the smallest AUC for the diagnosis of NS-TIs was for CRP (AUC = 0.762, 95% CI 0.667-0.857; P < 0.001). Additionally, according to the AUC value, the best combination was SAA and PCT for NS-PI diagnosis, and the best combination was PCT and IL-6 for NS-TI. In conclusion, compared with PCT and CRP, IL-6, IL-8, and SAA are better diagnostic biomarkers. Moreover, PCT combined with SAA is more suitable for diagnosing NS-PIs, and PCT combined with IL-6 is more suitable for diagnosing NS-TIs.

The COVID-19 pandemic has raised concerns regarding its potential impact on premature ovarian insufficiency (POI). This overview examines the possible interactions between COVID-19 and POI, while also suggesting preventive measures. The viral infection's inflammatory response and immune dysregulation may adversely affect ovarian tissues, leading to inflammation and damage. Additionally, alterations in vascular function could impair ovarian blood flow and hormonal imbalances may disrupt normal ovarian function. Long-term health effects, such as "long COVID," may exacerbate these issues through chronic inflammation and immune dysfunction. Public health measures, such as vaccination and home isolation, may indirectly protect ovarian health by reducing systemic inflammation. Vaccines could mitigate the severity of COVID-19's impact on ovarian function, while isolation may reduce stress and inflammation. However, further research is needed to validate these mechanisms.

Rafflesiaceae is a family of endangered plants whose members are solely parasitic to the tropical grape vine Tetrastigma (Vitaceae). Currently, the genetics of their crosstalk with the host remains unexplored. In this study, we use homology-based in silico approaches to characterize micro-RNAs (miRNAs) expressed by Sapria himalayana and Rafflesia cantleyi from published omics data. Derived from secondary structures or hairpins, miRNAs are small regulators of gene expression. We found that some plant-conserved miRNA still exists in Rafflesiaceae. Out of 9 highly conserved miRNA families in plants, 7 families (156/157, 159/319, 160, 165/166, 171, 172, 390) were identified with a total of 22 variants across Rafflesiaceae. Some miRNAs were missing endogenous targets and may have evolved to target host miRNA, though this requires experimental verification. Rafflesiaceae miRNA promoters are mostly inducible by ethylene that mediates stress response in the host but could be perceived by the parasites as a signal for growth. This study provides evidence that certain miRNAs with ancient origins in land plants still exist in Rafflesiaceae, though some may have been coopted by parasites to target host genes.

Functional dyspepsia (FD) is a chronic disease that occurs in the gastroduodenal region and significantly impacts human health. Maslinic acid (MA), a pentacyclic triterpene acid, is the primary bioactive ingredient in Chinese medicinal herbs such as hawthorn, which exhibits beneficial impacts on the regulation of various disease progressions. However, the specific functions and associated pathways of MA in FD progression remain unclear and require further investigation. In this work, it was demonstrated that MA enhanced the cell viability of human gastric smooth muscle cells (HGSMCs). In addition, the mitochondrial dysfunctions induced by carbonyl cyanide 3-chlorophenylhydrazone (CCCP) were rescued after MA treatment. Furthermore, autophagy was increased following CCCP treatment, but this phenomenon was counteracted after MA treatment. The oxidative stress, elevated after CCCP treatment, was alleviated following MA addition. Finally, the AMPK/SIRT1 pathway was suppressed after CCCP stimulation but was re-activated after MA treatment. In conclusion, it was uncovered that MA accelerated HGSMC viability and improved mitochondrial function, inhibited autophagy, alleviated oxidative stress, and stimulated the AMPK/SIRT1 pathway. This discovery may offer new insight into the therapeutic effects of MA in FD progression.

Drought is a major abiotic stress in restricting the growth, development, and yield of maize. As a significant epigenetic regulator, small RNA also functions in connecting the transcriptional and post-transcriptional regulatory network. Further to help comprehending the molecular mechanisms underlying drought adaptability and tolerance of maize, an integrated multi-omics analysis of transcriptome, sRNAome, and degradome was performed on the seedling roots of an elite hybrid Zhengdan958 under drought stress. In this study, 2,911 genes, 32 conserved miRNAs, and 12 novel miRNAs showed a significantly differential expression under drought stress. Moreover, 6,340 target genes of 445 miRNAs were validated using degradome sequencing, forming 281 miRNA-mRNA pairs in control (CK) and drought-stressed (DS) library. These target genes were mainly involved in the plant hormone signal transduction and phenylpropanoid biosynthesis pathways. The integrated multi-omics analysis revealed that five DEmiRNA-mRNA pairs displayed negatively correlated expression patterns, which were also verified by qRT-PCR. Tissue-specific expression profile and regulatory network analysis revealed that miR528a/b-Zm00001d021850, miR408a/b-Zm00001d020794, and miR164e-Zm00001d003414 might be essential in root-specific drought stress response of maize Zhengdan958 seedlings. These worthwhile will promote the functional characterization of miRNA-mRNA modules response to drought stress, and potentially contribute to drought-resistance breeding of maize.

This study aimed to investigate the differential expression of the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway in relation to the Toll-like receptor (TLR)/nuclear factor κB (NF-κB) signaling pathway in an obese rat model. A total of 200 8-week-old male Wistar rats were randomly assigned to a control group (Ctrl, n = 40) and an observation group (Obs, n = 160), with obesity induced through a high-fat diet. Following modeling, the Obs group was further divided into a model group, a PI3K/AKT inhibition group, a TLR/NF-κB inhibition group, and a combined PI3K/AKT + TLR/NF-κB inhibition group, with 40 rats in each. Metabolic changes were assessed by monitoring the glucose infusion rate (GIR), as well as conducting an intraperitoneal glucose tolerance test (IPGTT) and an intraperitoneal insulin tolerance test (IPITT). Hematoxylin and eosin staining was utilized to observe morphological changes in adipose tissue, while Western blotting was employed to detect the expression levels of proteins associated with the PI3K/AKT and TLR/NF-κB signaling pathways in adipose tissue. The results indicated that the Obs group exhibited significantly higher blood glucose and insulin levels during the IPGTT and IPITT experiments compared to the Ctrl group (P < 0.05). Additionally, the GIR, as well as the expression levels of p-PI3K and p-AKT proteins in the Obs group, were significantly lower than those in the Ctrl group (P < 0.05). In both the PI3K/AKT inhibition group and the combined PI3K/AKT + TLR/NF-κB inhibition group, the expression of relevant proteins further declined (P < 0.05). These findings suggest that while a high-fat diet decreases the activity of the PI3K/AKT signaling pathway, it concurrently promotes inflammatory responses by upregulating the TLR-4 and NF-κB signaling pathways, indicating a critical role for these pathways in obesity-related metabolic abnormalities.

Breast cancer is a common malignant tumor of women. Ki67 is an important biomarker of cell proliferation. With the quantitative analysis, it is an important indicator of malignancy for breast cancer diagnosis. However, it is difficult to accurately and quantitatively evaluate the count of positive nucleus during the diagnosis process of pathologists, and the process is time-consuming and labor-intensive. In this work, we employed a quantitative analysis method of Ki67 in breast cancer based on deep learning approach. For the diagnosis of breast cancer, according to breast cancer diagnosis guideline, we first identified the tumor region of Ki67 pathological image, neglecting the non-tumor region in the image. Then, we detect the nucleus in the tumor region to determine the nucleus location information. After that, we classify the detected nucleuses as positive and negative according to the expression level of Ki67. According to the results of quantitative analysis, the proportion of positive cells is counted. Combining the above process, we design a breast Ki67 quantitative analysis pipeline. The Ki67 quantitative analysis system was assessed on the validation set. The Dice coefficient of the tumor region segmentation model was 0.848, the Average Precision index of the nucleus detection model was 0.817, and the accuracy of the nucleus classification model was 96.66%. Besides, in clinical independent sample experiment, the results show that the proposed breast Ki67 quantitative analysis system achieve excellent correlation with the diagnosis efficiency of doctors improved more than ten times and the overall consistency of diagnosis is intra-group correlation coefficient: 0.964. The research indicates that our quantitative analysis method of Ki67 in breast cancer has high clinical application value.