Objective
We aim to familiarize the application status of metagenomic sequencing in diagnosing pulmonary infections, to compare metagenomic sequencing with traditional diagnostic methods, to conclude ...the advantages and limitations of metagenomic sequencing, and to provide some advice for clinical practice and some inspiration for associated researches.
Data Sources
The data were obtained from peer‐reviewed literature, white papers, and meeting reports.
Results
This review focused on the applications of untargeted metagenomic sequencing in lungs infected by bacteria, viruses, fungi, chlamydia pneumoniae, Mycoplasma pneumoniae, parasites, and other pathogens. Compared with conventional diagnostic methods, metagenomic sequencing is better in detecting novel, rare, and unexpected pathogens and being applied in co‐infections. Meanwhile, it can also provide more comprehensive information about pathogens. However, metagenomic sequencing still has limitations. Also, the situations that should be applied in and how the results should be interpreted are discussed in this review.
Conclusion
Metagenomic sequencing improves efficiency to identify pathogens compared with traditional diagnostic methods and can be applied in clinical diagnosis. However, the technology of metagenomic sequencing still needs to be improved. Also, clinicians should learn more about when to use metagenomic sequencing and how to interpret its results.
Compared with conventional diagnostic methods, metagenomic sequencing is more sensitive and better in detecting novel, rare, and unexpected pathogens. Meanwhile, it can provide more comprehensive information about pathogens. However, many challenges exist when using metagenomic sequencing. By Figdraw (www.figdraw.com).
Transcobalamin (TCN1) is a vitamin B12 (cobalamin)-binding protein that regulates cobalamin homeostasis. Recent studies and bioinformatic analyses have found that TCN1 is highly expressed in cancer ...tissues and is associated with tumour aggressiveness and poor prognosis. The present study aimed to detect TCN1 as a novel biomarker for prognosis and chemosensitivity of colon cancer. Next-generation sequencing showed that TCN1 was one of several upregulated mRNAs in colon cancer, which was verified by further bioinformatics analyses. Western blotting (n = 9) and quantitative real time polymerase chain reaction (qRT-PCR, n = 30) revealed that TCN1 was highly expressed in colon cancer tissues at both the protein and mRNA level. A total of 194 cases of colon cancer were examined by immunohistochemistry and revealed that TCN1 expression level was related to advanced stages (P < 0.005). Kaplan-Meier analysis verified that patients with lower TCN1 expression usually had longer overall survival (P = 0.008). In addition, TCN1 was highly expressed in pulmonary metastatic tumour tissues (n = 37, P = 0.025) and exhibited higher levels in right-sided colon cancer than in left-sided colon cancer (P = 0.029). TCN1 expression in specimens that had received neoadjuvant chemotherapy decreased compared with that in colonoscopy biopsy tissues (n = 42, P = 0.009). Further bioinformatics analyses verified that apoptosis pathways might have a role in high TCN1 expression. All the studies revealed that TCN1 expression in colon cancer was significantly associated with malignant biological behaviour. Therefore, TCN1 could be used as a novel biomarker for colon cancer aggressiveness and prognosis and might also be a potential biomarker for predicting neoadjuvant chemosensitivity.
The ionic liquid of 1-allyl-3-methylimidazolium chloride (amimCl) was used as the good solvent to dissolve celluloses. Cellulose concentration covers the range of 0.1−3.0 wt %, spanning both the ...dilute and semidilute regimes. The rheological properties of the cellulose ionic liquid solutions have been investigated by steady shear and oscillatory shear measurements in this study. In the steady shear measurements, all the cellulose solutions show a shear thinning behavior at high shear rates; however, the dilute cellulose solutions show another shear thinning region at low shear rates, which may reflect the characteristics of the amimCl solvent. In the oscillatory shear measurements, for the dilute regime, the reduced dimensionless moduli are obtained by extrapolation of the viscoelastic measurements for the dilute solutions to infinite dilution. The frequency dependences of the reduced dimensionless moduli are intermediate between the predictions from the Zimm model and elongated rodlike model theories, while the fitting by using a hybrid model combining these two model theories agrees well with the experimental results. For the semidilute regime, the frequency dependences of moduli change from the Zimm-like behavior to the Rouse-like behavior with increasing cellulose concentration. In the studied concentration range, the effects of molecular weight and temperature on solution viscoelasticities and the relationship between steady shear viscosity and dynamic shear viscosity are presented. Results show that the solution viscoelasticity greatly depends on the molecular weight of cellulose; the empirical time−temperature superposition principle holds true at the experimental temperatures, while the Cox−Merz rule fails for the solutions investigated in this study.
Unrestrained activation of Th1 and Th17 cells is associated with the pathogenesis of multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis (EAE). While inactivation of ...dynamin-related protein 1 (Drp1), a GTPase that regulates mitochondrial fission, can reduce EAE severity by protecting myelin from demyelination, its effect on immune responses in EAE has not yet been studied.
We investigated the effect of Mdivi-1, a small molecule inhibitor of Drp1, on EAE. Clinical scores, inflammation, demyelination and Drp1 activation in the central nervous system (CNS), and T cell responses in both CNS and periphery were determined.
Mdivi-1 effectively suppressed EAE severity by reducing demyelination and cellular infiltration in the CNS. Mdivi-1 treatment decreased the phosphorylation of Drp1 (ser616) on CD4
T cells, reduced the numbers of Th1 and Th17 cells, and increased Foxp3
regulatory T cells in the CNS. Moreover, Mdivi-1 treatment effectively inhibited IFN-γ
, IL-17
, and GM-CSF
CD4
T cells, while it induced CD4
Foxp3
regulatory T cells in splenocytes by flow cytometry.
Together, our results demonstrate that Mdivi-1 has therapeutic potential in EAE by modulating the balance between Th1/Th17 and regulatory T cells.
Emerging evidence suggests an association of Alzheimer's Disease (AD) with microglial and astrocytic dysregulation. Recent studies have proposed that activated microglia can transform astrocytes to a ...neurotoxic A1 phenotype, which has been shown to be involved in the promotion of neuronal damage in several neurodegenerative diseases, including AD. In the present study, we observed an obvious microglial activation and A1-specific astrocyte response in the brain tissue of APP/PS1 Tg mice. Fasudil treatment improved the cognitive deficits of APP/PS1 Tg mice, inhibited microglial activation and promoted their transformation to an anti-inflammatory phenotype, and further shifted astrocytes from an A1 to an A2 phenotype. Our experiments suggest Fasudil exerted these functions by inhibing the expression of TLR4, MyD88, and NF-κB, which are key mediators of inflammation.
Using in vitro experiments, we further validated in vivo findings. Our cell experiments indicated that Fasudil induces a shift of inflammatory microglia towards an anti-inflammatory phenotype. LPS-induced microglia-conditioned medium promotes A1 astrocytic polarization, but Fasudil treatment resulted in a direct transformation of A1 astrocytes to A2. To summarize, our results show that Fasudil inhibits the neurotoxic activation of microglia and shifts astrocytes towards a neuroprotective A2 phenotype, representing a promising candidate for AD treatment.
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•Fasudil could inhibit the activation of microglia in the brain tissue of APP/PS1 Tg mice.•Fasudil could shift astrocytes from an A1 to an A2 phenotype in the brain tissue of APP/PS1 Tg mice.•Fasudil could reverse microglial-induced neurotoxic astrocyte(A1) to A2 in vitro.•The study provides preclinical evidence that Fasudil treatment ameliorated memory deficits in APP/PS1 Tg mice.
The outbreak of COVID-19 has led to international concern. We aimed to establish an effective screening strategy in Shanghai, China, to aid early identification of patients with COVID-19.
We did a ...multicentre, observational cohort study in fever clinics of 25 hospitals in 16 districts of Shanghai. All patients visiting the clinics within the study period were included. A strategy for COVID-19 screening was presented and then suspected cases were monitored and analysed until they were confirmed as cases or excluded. Logistic regression was used to determine the risk factors of COVID-19.
We enrolled patients visiting fever clinics from Jan 17 to Feb 16, 2020. Among 53 617 patients visiting fever clinics, 1004 (1·9%) were considered as suspected cases, with 188 (0·4% of all patients, 18·7% of suspected cases) eventually diagnosed as confirmed cases. 154 patients with missing data were excluded from the analysis. Exposure history (odds ratio OR 4·16, 95% CI 2·74–6·33; p<0·0001), fatigue (OR 1·56, 1·01–2·41; p=0·043), white blood cell count less than 4 × 109 per L (OR 2·44, 1·28–4·64; p=0·0066), lymphocyte count less than 0·8 × 109 per L (OR 1·82, 1·00–3·31; p=0·049), ground glass opacity (OR 1·95, 1·32–2·89; p=0·0009), and having both lungs affected (OR 1·54, 1·04–2·28; p=0·032) were independent risk factors for confirmed COVID-19.
The screening strategy was effective for confirming or excluding COVID-19 during the spread of this contagious disease. Relevant independent risk factors identified in this study might be helpful for early recognition of the disease.
National Natural Science Foundation of China.
Summary
Aim
Multiple sclerosis (MS) is a relapsing‐remitting inflammatory demyelinating disease that requires long‐term treatment. Although Rho kinase inhibitor Fasudil shows good therapeutic effect ...in experimental autoimmune encephalomyelitis (EAE), an animal model of MS, certain side effects may limit its clinical use. This study aimed at observing the therapeutic potential of Fasudil‐modified encephalitogenic mononuclear cells (MNCs) via nasal delivery in EAE and exploring possible mechanisms of action.
Methods
Experimental autoimmune encephalomyelitis was induced with myelin oligodendrocyte glycoprotein 35‐55 in C57BL/6 mice, and encephalitogenic MNCs were treated with Fasudil in vitro. Mice received 3 × 106 cells/10 μL per nasal cavity on day 3 and 11 postimmunization, respectively.
Results
Fasudil‐modified MNCs reduced clinical severity of EAE, improved demyelination, and decreased inflammatory cells in spinal cords. Immunohistochemical results indicated that CD4+ T cells and CD68+ macrophages were barely detected in Fasudil‐MNCs group. Fasudil‐modified MNCs decreased CD4+IFN‐γ+ and CD4+IL‐17+ T cells, increased CD4+IL‐10+ T cells, restrained M1 markers CD16/32, CCR7, IL‐12, CD8a, enhanced M2 markers CD206, CD200, CD14 in spleen. Fasudil‐modified MNCs inhibited the activation of inflammatory signaling p‐NF‐kB/P38, accompanied by the decrease of COX‐2 and the increase of Arg‐1 in spinal cord, as well as the reduction of IL‐17, TNF‐α, IL‐6 and the elevation of IL‐10 in cultured supernatant of splenocytes. Fasudil‐modified MNCs enhanced the levels of neurotrophic factors BDNF and NT‐3 in spinal cord.
Conclusion
Our results indicate that intranasal delivery of Fasudil‐modified MNCs have therapeutic potential in EAE, providing a safe and effective cell therapeutic strategy to MS and/or other related disorders.
To prospectively assess the feasibility of using virtual iron content (VIC) imaging at dual-energy computed tomography (CT) to evaluate the liver iron content (LIC) in patients suspected of having ...liver iron overload and to compare the LIC grading performance of VIC imaging and magnetic resonance (MR) imaging.
This study was approved by the institutional review board, and informed consent was obtained from all patients. Fifty-six patients suspected of having liver iron overload (serum ferritin concentrations >500 μg/L) underwent unenhanced dual-energy CT and MR imaging of the liver. MR imaging-measured LICs were obtained in 34 of the 56 patients. VIC images were generated with dual-energy analysis. R2* and MR-measured LIC were obtained with gradient-echo and spin-echo sequences, respectively. Correlations between CT and MR measurements were analyzed. The diagnostic performance of VIC and R2* in the differentiation of different LIC thresholds were evaluated with receiver operating characteristic (ROC) analysis.
Hepatic VIC showed significant correlation with R2* and MR-measured LIC (r = 0.885 and 0.871, respectively; P < .0001). To differentiate among different LIC thresholds of 1.8, 3.2, 7.0, and 15.0 mg of iron per gram of dry tissue, the corresponding optimal cutoff values for VIC were 2.50, 5.13, 8.93, and 17.97 HU, respectively. At a LIC threshold of 7.0 mg of iron per gram of dry tissue or higher, 100% sensitivity (15 of 15 patients) and 100% specificity (19 of 19 patients) were obtained for VIC. There was no significant difference between VIC and R2* (area under the ROC curve, 0.964 vs 0.993, respectively; P = .299) in grading LIC levels at a LIC threshold of 3.2 mg of iron per gram of dry tissue or higher. Conclusion Hepatic VIC is a potential index for accurately evaluating and grading clinically significant liver iron accumulation, with a diagnostic performance similar to that of MR imaging.
Although Fasudil has shown therapeutic potential in EAE mice, the mechanism of action are still not fully understood. Here, we examined the immunomodulatory effect of Fasudil on encephalitogenic ...mononuclear cells (MNCs), and tested the therapeutic potential of Fasudil‐treated MNCs in active EAE. Fasudil inhibited expression of CCL20 on T cells and migration of T cells, decreased CD4+IFN‐γ+ and CD4+IL‐17+ T cells, but increased CD4+IL‐10+ and CD4+TGF‐β+ T cells. Fasudil reduced expression of CD16/32 and IL‐12, while elevating expression of CD206, CD23, and IL‐10. Fasudil also decreased levels of iNOS/NO, enhanced levels of Arg‐1, and inhibited the TLR‐4/NF‐κB signaling and TNF‐α, shifting M1 macrophage to M2 phenotype. These modulatory effects of Fasudil on T cells and macrophages were not altered by adding autoantigen MOG35–55 to the culture, i.e., autoantigen‐independent. Further, we observed that, in vitro, Fasudil inhibited the capacity of encephalitogenic MNCs to adoptively transfer EAE and reduced TLR‐4/p‐NF‐κB/p65 and inflammatory cytokines in spinal cords. Importantly, Fasudil‐treated encephalitogenic MNCs exhibited therapeutic potential when injected into actively induced EAE mice. Together, our results not only provide evidence that Fasudil mediates the polarization of macrophages and the regulation of T cells, but also reveal a novel strategy for cell therapy in MS.
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