StAP1 phage: an effective tool for treating methicillin-resistant Staphylococcus aureus infections.

Introduction: Staphylococcus aureus infection has long been a serious concern in the medical field, with methicillin-resistant Staphylococcus aureus (MRSA) posing a considerable challenge to public health. Given the escalating bacterial resistance and the favorable biosafety and environmental properties of phages, the resurgence of phage therapy offers a promising alternative to antibiotics. Methods: In this study, we isolated and characterized a MRSA phage named StAP1 from a Chinese hospital. Phenotypic and molecular analyses revealed its broad-spectrum characteristics, genomic background, and potential application in MRSA infection treatment. Results: Morphological examination classified the phage as a member of the Herelleviridae phage family, displaying a typical hexagonal head and a slender fibrous tail. Genomic analysis unveiled a size of ~144,705 bp for the StAP1 genome, encompassing 215 open reading frames (ORFs). The one-step growth curve demonstrated a 20-min incubation period for the phage, with an optimal multiplicity of infection (MOI) of 0.1. Moreover, StAP1 exhibited stability across a wide range of temperatures and pH levels. Further investigation of its broad-spectrum characteristics confirmed its ability to effectively infect all staphylococcal cassette chromosomal mec (SCCmec) types found in MRSA strains, notably displaying a remarkable lysis rate of 76.7% against the prevalent ST239 strain in China. In vivo studies show cased significant efficacy of the StAP1 phage against MRSA infection. Discussion: Overall, StAP1 phage presents a broad infection spectrum and exhibits strong lytic effects on various MRSA strains, highlighting its tremendous potential as a powerful tool for MRSA infection treatment.

Interrelation of Chronic Lung Disease and Cardiovascular Disease Based on Two National Prospective Cohort Studies.

BACKGROUND: Chronic lung diseases (CLDs) and cardiovascular diseases (CVDs) are the main chronic diseases responsible for a considerable burden of disease. This study aimed to estimate the interrelation of CLDs and CVDs using two Chinese national longitudinal cohort studies. METHODS: The China Health and Retirement Longitudinal Study (CHARLS) and Chinese Longitudinal Healthy Longevity Survey (CLHLS) were used in this study with 15,052 and 9,765 participants, respectively. The Cox proportional risk model was used to estimate the interrelation between CLDs and CVDs. Mediating effects were performed to detect possible influencing pathways between CLDs and CVDs. RESULTS: The association of CLDs with CVDs was identified in 1,647 participants (10.9%) with newly diagnosed CVDs in CHARLS and 332 participants (11.6%) in CLHLS. The Cox proportional risk model showed that CLDs were a significant predictor of CVDs (HR 1.49, 95% CI 1.27-1.76) after adjusting for covariates, and the hazard ratios of stroke and CVDs excluding stroke were (HR 1.02, 95% CI 0.79-1.31) and (HR 1.76, 95% CI 1.46-2.13), respectively. These association were mediated by body mass index (BMI) and Center for Epidemiological Studies Depression Scale (CES-D-10) scores. No significant association was found in CHARLS and CLHLS regarding CVDs with CLDs. In CHARLS, CVDs was a significant predictor of CLDs (HR 1.40, 95% CI 1.09-1.79). CONCLUSIONS: Chronic lung disease was associated with increased incidence of CVDs in middle-aged and older people in the community population and vice versa. Body mass index and depressive symptoms might be mediated by the effect of CLD on CVD.

[Application of health failure mode and effect analysis for the airbag pressure management of patients with artificial airways].

OBJECTIVE: To analyze the application effect of health failure mode and effect analysis (HFMEA) model in patients with artificial airways in the cardiovascular surgery intensive care unit (CSICU) by establishing a HFMEA project team, and to develop targeted improvement measures and processes. METHODS: The patients undergoing cardiovascular surgeries and with established artificial airways in the Shandong Provincial Hospital Affiliated to Shandong First Medical University were recruited from October 2021 to March 2022. The enrolled patients were divided into the conventional management group and the HFMEA model management group according to random number table method. The conventional management group applied the conventional procedures for monitoring the air bag pressure. The HFMEA model management group used the HFMEA model to implement and improve the airbag pressure monitoring process. The efficacy of HFMEA was assessed by comparing the incidence of ventilator-associated pneumonia (VAP), the pass rate of airbag pressure monitoring, the duration of endotracheal intubation and the length of CSICU stay between two groups. The practicability of HFMEA model was evaluated by analyzing the theoretical assessment scores and practical skill scores of nurses and their satisfaction scores with HFMEA. RESULTS: Compared with the conventional management group, the patients in the HFMEA mode management group had a significantly higher rate of passing airbag pressure monitoring [94.99% (2 994/3 152) vs. 69.97% (1 626/2 324), P < 0.01], shorter duration of endotracheal intubation and length of CSICU stay [duration of endotracheal intubation (hours): 6 (7, 12) vs. 6 (8, 13), length of CSICU stay (hours): 40 (45, 65) vs. 41 (46, 85), both P < 0.05], but the incidences of VAP between the two groups were similar. The theoretical assessment scores and practical skill scores of nurses were significantly higher (theoretical assessment score: 44.47±2.72 vs. 37.59±6.56, practical skill score: 44.56±2.66 vs. 40.03±4.32, total score: 89.03±3.07 vs. 77.63±9.56, all P < 0.05) in the HFMEA mode management group. And the satisfaction scores with airbag pressure management were also significantly higher in the HFMEA mode management group (7.72±1.11 vs. 6.44±1.32, P < 0.05). CONCLUSIONS: The application of the HFMEA can improve the airbag pressure measures and standardize the monitoring procedures in patients with artificial airways, and reduce the risk of clinical nursing. It is safe and effective for patients with invasive mechanical ventilation in the CSICU.

Prone positioning in extubated patients with hypoxemic respiratory failure after cardiac surgery: A retrospective study.

BACKGROUND: Hypoxemic respiratory failure is a serious complication that can occur at any stage after cardiac surgery. Prone positioning (PP) is safe and effective for patients receiving invasive ventilation after hypoxemic respiratory failure; however, few related studies have focused on its use with extubated cardiac surgery patients. Researchers recently reported beneficial effects of PP for hypoxemic patients with COVID-19 and those with moderate ARDS (acute respiratory distress syndrome,ARDS). PP may also improve oxygenation in extubated cardiac surgery patients. OBJECTIVE: In this study, we aimed to assess the safety and effectiveness of PP in extubated cardiac surgery patients to determine whether PP can improve oxygenation and respiratory status or reduce secondary intubation. METHODS: We reviewed our institutional database between August 2018 and August 2020 and identified 22 cardiac surgery patients who had undergone PP for hypoxemic respiratory failure after extubation. From the medical and nursing records, we extracted the following data recorded before PP, during PP, and after PP for each patient, arterial blood gas analyses, hemodynamic records, laboratory reports, and respiratory function training records. RESULTS: Twenty-two extubated patients underwent 74 PP. Each patient underwent a median of 3.5 (2-5) procedures, and the median duration of each PP was 10 h. PP was implemented on the 4.5th postoperative day (median). All patients were discharged from the hospital, and none died. No complications were observed. PP improved the P/F ratio (182.65 ± 60.17, 301.53 ± 61.31, and 246.76 ± 65.68, before PP, during PP, and after PP, respectively, p < 0.001). Additionally, the respiratory rate, Forced Vital Capacity (FVC) and PaCO2 also improved, and hemodynamics showed no significant change. CONCLUSION: PP may be effective and safe for treating patients who are extubated following cardiac surgery with hypoxemic respiratory failure. For these patients, PP is associated with oxygenation and respiratory condition improvements and low secondary intubation rates.

Impact of a sleep promotion protocol on off-pump coronary artery bypass graft patients.

BACKGROUND: Sleep abnormalities frequently occur in intensive care unit (ICU) patients, and the consequences of sleep abnormalities in patients who undergo off-pump coronary artery bypass graft (OPCABG) surgery are particularly significant. Although many interventions have been reported to improve sleep, few sleep promotion protocols have been designed specifically for patients in cardiac ICUs. AIMS AND OBJECTIVES: This study aimed to explore the effects of an evidence-based sleep promotion protocol on patients who underwent OPCABG in a cardiac ICU. DESIGN: A quasi-experimental study was conducted in a comprehensive hospital in Shandong province of China. METHODS: Overall, 67 participants were recruited (37 in the control group and 30 in the intervention group). An evidence-based sleep promotion protocol was developed by a 10-member interprofessional collaborative team and then applied. Sound levels, light intensity, and the number of nocturnal interventions were compared between groups. The Chinese version of the Richards-Campbell Sleep Questionnaire (RCSQ) was used to compare intergroup sleep status on two consecutive postoperative nights. RESULTS: No significant differences were found for demographics or disease severity between the groups. In the intervention group, sound levels and light intensity were significantly lower at various times, and nocturnal interventions were significantly less frequent over the two consecutive nights. RCSQ scores were significantly higher in the intervention group for both nights. CONCLUSIONS: The sleep promotion protocol reduced sound levels, night-time light intensity, the number of nocturnal interventions, and improved sleep among OPCABG patients in a cardiac ICU. RELEVANCE TO CLINICAL PRACTICE: Evidence-based practice can help to promote good quality of care, improve patient outcomes, and advance nursing in clinical settings.

Integrated mRNA-seq and miRNA-seq analysis of goat fibroblasts response to Brucella Melitensis strain M5-90.

Brucellosis is a globally zoonotic bacterial disease of humans and various animals including goats, sheep, and cattle. Brucella melitensis M5-90, a live attenuated vaccine strain, has been widely used to prevent brucellosis in goats and sheep. However, the molecular mechanisms governing protective immunity response in non-professional phagocytes infected with B. melitensis M5-90 have not been fully investigated, especially in goats. In our research, goat fibroblasts were used as in vitro models to determine these mechanisms by transcriptome analysis. After incubating with B. melitensis M5-90 3 h, the infected goat fibroblasts were collected at 0 h, 4 h, 24 h, 48 h and 72 h for RNA-seq. The results indicated that there were totally 11,819 differentially expressed genes (DEGs) and 777 differentially expressed (DE) miRNAs found in experiment groups compared with the control groups (|log2(Foldchange)|≥1, FDR<0.05). GO and KEGG enrichment analyses revealed that down-regulated genes were involved in the riboflavin metabolism and positive regulation of IL-8 secretion pathway. The up-regulated genes were mainly involved in adaptive immunity, including TNF signaling pathway, MAPK signaling pathway and JAK/STAT pathway. Additionally, cytokine-cytokine receptor interaction, natural killer cell mediated cytotoxicity and toll-like receptor signaling pathway, which associated with innate immunity pathways, were also induced. Based on the Pearson correlation coefficients and prediction results of TargetScan and miRanda, the miRNA-mRNA networks of NFKB1, IFNAR2 and IL10RB were constructed and verified in goat fibroblasts by qPCR, which demonstrated that goat fibroblasts displayed immunomodulatory properties. Our findings provide a deeper insight into the host miRNA-driven B. melitensis defense mechanism and reveal the transcriptome changes involved in the innate and adaptive immune response of the goats to B. melitensis infection.

miR-146b-5p Plays a Critical Role in the Regulation of Autophagy in ∆per Brucella melitensis-Infected RAW264.7 Cells.

Brucella-caused brucellosis is one of the most widespread worldwide zoonoses. Lipopolysaccharide (LPS) of Brucella, which functions as pathogen-associated molecular patterns (PAMPs), is an important virulence factor that elicits protective antibodies. Per of B. melitensis is involved in the biosynthesis of the O-side chain of LPS. Autophagy is a crucial element of the innate immune response against intracellular pathogens including Brucella. In this study, we observed that autophagy was inhibited in RAW264.7 cells infected with Brucella melitensis ∆per. And, a high-throughput array-based screen and qRT-PCR validation were performed to identify the differentially expressed miRNAs in RAW264.7 cells infected with B. melitensis M5-90 ∆per. The results suggested that mmu-miR-146a-5p, mmu-miR-155-5p, mmu-miR-146b-5p, and mmu-miR-3473a were upregulated and mmu-miR-30c-5p was downregulated. During B. melitensis M5-90 ∆per infection, the increased expression of miR-146b-5p inhibited the autophagy activation in RAW264.7 cells. Using a bioinformatics approach, Tbc1d14 was predicted to be a potential target of miR-146b-5p. The results of a luciferase reporter assay indicated that miR-146b-5p directly targeted the 3'-UTR of Tbc1d14, and the interaction between miR-146b-5p and the 3'-UTR of Tbc1d14 was sequence-specific. High-throughput RNA-Seq-based screening was performed to identify differentially expressed genes in Tbc1d14-expressing RAW264.7 cells, and these were validated by qRT-PCR. Among the differentially expressed genes, four autophagy associated genes, IFNγ-inducible p47 GTPase 1 (IIGP1), nuclear receptor binding protein 2 (Nrbp2), transformation related protein 53 inducible nuclear protein 1 (Trp53inp1), and immunity-related GTPase family M member 1 (Irgm1), were obtained. Our findings provide important insights into the functional mechanism of LPS of B. melitensis.

Integrated mRNA and miRNA profiling in NIH/3T3 cells in response to bovine papillomavirus E6 gene expression.

Delta bovine papillomaviruses (δBPVs) mainly infect cattle and cause fibropapillomas. δBPVs encode three oncogenes, E5, E6 and E7. The effect of E6 on microRNA (miRNA) and mRNA expression profiles is not well characterized. In this study, RNA sequencing and small RNA sequencing were used to explore alterations in mRNAs and miRNAs in E6 over-expressing NIH/3T3 cells (NH-E6) compared with control cells (NH-GFP). We found that 350 genes (181 upregulated and 169 downregulated) and 54 miRNAs (26 upregulated and 28 downregulated) were differentially expressed (DE) following E6 expression. The top 20 significantly enriched GO terms in "biological process" included inflammatory response, innate immune response, immune response, immune system process, positive regulation of apoptotic process, cell adhesion, and angiogenesis. We constructed a potential miRNA-gene regulatory network from the differentially expressed genes (DEGs) and DE miRNAs. Finally, we selected 19 immune-response related DEGs and 11 DE miRNAs for qPCR validation. Of these, upregulation of 12 genes, Ccl2, Ccl7, Cxcl1, Cxcl5, Tlr2, Nfkbia, Fas, Il1rl1, Ltbp1, Rab32, and Zc3h12a, Dclk1 and downregulation of four genes, Agtr2, Ptx3, Sfrp1, and Thbs1 were confirmed. Ccl2, Ccl7, Cxcl1 and Cxcl5 were upregulated more than ten-fold in NH-E6 compared with NH-GFP. Also, upregulation of three miRNAs, mmu-miR-129-2-3p, mmu-miR-149-5p-R-2 and mmu-miR-222-3p, and downregulation of five miRNAs, mmu-miR-582-3p-R+1, mmu-miR-582-5p, mmu-miR-708-3p, mmu-miR-708-5p and mmu-miR-1197-3p, were confirmed. Our study describes changes in both mRNA and miRNA profiles in response to BPV E6 expression, providing new insights into BPV E6 oncogene functions.

Enhanced luminescence properties of InAs nanowires via organic and inorganic sulfide passivation.

InAs nanowires (NWs) have been considered to be highly suitable for future nanoscale photonic applications in mid-wave infrared region. However, progress in this area has been seriously hampered because of the poor radiative efficiency of InAs NWs attributed to their non-radiative (NR) surface. Herein, we demonstrated that a significant improvement of optical performances of InAs NWs grown by chemical vapor deposition could be achieved via sulfur passivation process. Luminescence properties of InAs NWs via organic sulfide (ODT) and inorganic sulfide ((NH4)2S) passivation were investigated by detailed photoluminescence (PL) measurement, which exhibited ∼17-fold enhancement in the intensity of optical emission compared to unpassivated InAs NWs. Moreover, the results of this investigation revealed that compared to ODT passivation, (NH4)2S passivation provided a more effective method to enhance the luminescence intensity even up to room temperature. This improvement of optical emission arises from the efficient passivation of surface defect states which act as competing NR centers. Furthermore, the stability of the passivated InAs NWs was investigated by PL measurement as a function of storage time in air. These findings are important for the successful implementation of optoelectronic devices based on InAs NWs.

Genome-wide analysis of circular RNAs in goat skin fibroblast cells in response to Orf virus infection.

Orf, caused by Orf virus (ORFV), is a globally distributed zoonotic disease responsible for serious economic losses in the agricultural sector. However, the mechanism underlying ORFV infection remains largely unknown. Circular RNAs (circRNAs), a novel type of endogenous non-coding RNAs, play important roles in various pathological processes but their involvement in ORFV infection and host response is unclear. In the current study, whole transcriptome sequencing and small RNA sequencing were performed in ORFV-infected goat skin fibroblast cells and uninfected cells. A total of 151 circRNAs, 341 messenger RNAs (mRNAs), and 56 microRNAs (miRNAs) were differently expressed following ORFV infection. Four circRNAs: circRNA1001, circRNA1684, circRNA3127 and circRNA7880 were validated by qRT-PCR and Sanger sequencing. Gene ontology (GO) analysis indicated that host genes of differently expressed circRNAs were significantly enriched in regulation of inflammatory response, epithelial structure maintenance, positive regulation of cell migration, positive regulation of ubiquitin-protein transferase activity, regulation of ion transmembrane transport, etc. The constructed circRNA-miRNA-mRNA network suggested that circRNAs may function as miRNA sponges indirectly regulating gene expression following ORFV infection. Our study presented the first comprehensive profiles of circRNAs in response to ORFV infection, thus providing new clues for the mechanisms of interactions between ORFV and the host.

Alterations of microRNAs and their predicted targeting mRNAs expression in RAW264.7 macrophages infected with Omp25 mutant Brucella melitensis.

Brucellosis is a worldwide zoonosis caused by Brucella species and represents a serious threat to both human and animal health. Omp25 is an important immunogenic and protective antigen in Brucella species; however, the functional mechanism of Omp25 in macrophages has not yet been elucidated. Here, we constructed a Brucella melitensis omp25 deletion mutant (M5-90-Δ omp25) and performed microRNA (miRNA) profiling of infected RAW264.7 cells. Eight differentially expressed miRNAs ( mmu-miR-146a-5p, mmu-miR-155-5p, mmu-miR-3473a, mmu-miR-149-3p, mmu-miR-671-5p, mmu-miR-1224-5p, mmu-miR-1895, and mmu-miR-5126) were identified, with quantitative real-time PCR (qRT-PCR) analysis confirming the up-regulation of mmu-miR-146-a-5p and mmu-miR-155-5p and down-regulation of mmu-miR-149-3p and mmu-miR-5126. mRNA profiling of B. melitensis M5-90-Δo mp25-infected RAW264.7 cells identified 967 differentially expressed genes (DEGs) (fold change ≥ 2). Among these, we focused on genes that were predicted by TargetScan, miRanda, and PicTar to be the potential targets of the differentially expressed miRNAs. The results suggested that 17 separate genes are potentially targeted by mmu-miR-149-3p, with one of these genes, Tbr1, also targeted by mmu-miR-5126. qRT-PCR analysis confirmed the up-regulation of nine of the predicted target genes. Our findings provide important information about the functional molecules in host cells, including miRNA and their target genes, affected by Omp25 from Brucella. This information is particularly valuable for the prophylaxis and treatment of brucellosis.

Upregulation of Immune Process-Associated Genes in RAW264.7 Macrophage Cells in Response to Burkholderia pseudomallei Infection.

Melioidosis is a severe and fatal tropical zoonosis, which is triggered by Burkholderia pseudomallei. To better understand the host's response to infection of B. pseudomallei, an RNA-Seq technology was used to confirm differentially expressed genes (DEGs) in RAW264.7 cells infected with B. pseudomallei. In total, 4668 DEGs were identified across three time points (4, 8, and 11 hours after infection). Short Time-Series Expression Miner (STEM) analysis revealed the temporal gene expression profiles and identified seven significant patterns in a total of 26 profiles. Kyoto Encyclopedia of Genes and Genomes (KEGG) was utilized to confirm significantly enriched immune process-associated pathways, and 10 DEGs, including Ccl9, Ifnb1, Tnfα, Ptgs2, Tnfaip3, Zbp1, Ccl5, Ifi202b, Nfkbia, and Nfkbie, were mapped to eight immune process-associated pathways. Subsequent quantitative real-time PCR assays confirmed that the 10 DEGs were all upregulated during infection. Overall, the results showed that B. pseudomallei infection can initiate a time-series upregulation of immune process-associated DEGs in RAW264.7 macrophage cells. The discovery of this article helps us better understand the biological function of the immune process-associated genes during B. pseudomallei infection and may aid in the development of prophylaxis and treatment protocols for melioidosis.