Trojan-horse inspired nanoblaster: X-ray triggered spot attack on radio-resistant cancer through radiodynamic therapy.

Radiotherapy as a mainstay of in-depth cervical cancer (CC) treatment suffers from its radioresistance. Radiodynamic therapy (RDT) effectively reverses radio-resistance by generating reactive oxygen species (ROS) with deep tissue penetration. However, the photosensitizers stimulated by X-ray have high toxicity and energy attenuation. Therefore, X-ray responsive diselenide-bridged mesoporous silica nanoparticles (DMSNs) are designed, loading X-ray-activated photosensitizer acridine orange (AO) for spot blasting RDT like Trojan-horse against radio-resistance cervical cancer (R-CC). DMSNs can encapsulate a large amount of AO, in the tumor microenvironment (TME), which has a high concentration of hydrogen peroxide, X-ray radiation triggers the cleavage of diselenide bonds, leading to the degradation of DMSNs and the consequent release of AO directly at the tumor site. On the one hand, it solves the problems of rapid drug clearance, adverse distribution, and side effects caused by simple AO treatment. On the other hand, it fully utilizes the advantages of highly penetrating X-ray responsive RDT to enhance radiotherapy sensitivity. This approach results in ROS-induced mitochondria damage, inhibition of DNA damage repair, cell cycle arrest and promotion of cancer cell apoptosis in R-CC. The X-ray responsive DMSNs@AO hold considerable potential in overcoming obstacles for advanced RDT in the treatment of R-CC.

Development and application of visual fish freshness indicator film incorporated with anthocyanins encapsulation by whey protein-propylene glycol alginate nanoparticles.

Intelligent indicator packaging has gained increased attention in meeting the demand for reducing food waste and mitigating the risk of food poisoning. This study focused on the preparation, characteristics, and application of freshness indicator films based on polyvinyl alcohol (PVA) and anthocyanins (ACNs)-encapsulated whey protein (WP)-propylene glycol alginate (PGA) nanoparticles. The successful encapsulation of ACNs by WP-PGA nanoparticles improved the stability of ACNs and their encapsulation efficiency (EE) reached 95.34 %. The incorporation of nanoparticles into the structure of the PVA films was justified by SEM, XRD, and ATR-FTIR, and resulted in a decrease in water vapor permeability (WVP) from 15.76 (×10-7 g·m-1·Pa-1·h-1) to 8.40 (×10-7 g·m-1·Pa-1·h-1) and an increase in scavenging rate of DPPH radical from 1.47 % to 18.92 %. The light-blocking property and mechanical properties of the films were also improved, and they showed visible color change in response to pH 2-12 and high color stability after 14 days of storage at 4 °C and 25 °C. Furthermore, the freshness indicator films underwent a noticeable transformation from rosy red to gray at bighead carp head spoilage. Therefore, the encapsulation of ACNs using WP-PGA nanoparticles provides a promising non-destructive and real-time freshness indication of aquatic products during both storage and transportation processes.

Accuracy of cell-free Mycobacterium tuberculosis DNA testing in pleural effusion for diagnosing tuberculous pleurisy: a multicenter cross-sectional study.

BACKGROUND: The diagnosis of tuberculous pleurisy (TP) presents a significant challenge due to the low bacterial load in pleural effusion (PE) samples. Cell-free Mycobacterium tuberculosis DNA (cf-TB) in PE samples is considered an optimal biomarker for diagnosing TP. This study aimed to evaluate the applicability of cf-TB testing across diverse research sites with a relatively large sample size. METHODS: Patients suspected of TP and presenting with clinical symptoms and radiological evidence of PE were consecutively enrolled by treating physicians from 11 research sites across 6 provinces in China between April 2020 and August 2022. Following centrifugation, sediments obtained from PE were used for Xpert MTB/RIF (Xpert) and mycobacterial culture, while the supernatants were subjected to cf-TB testing. This study employed a composite reference standard to definite TP, which was characterized by any positive result for Mycobacterium tuberculosis (MTB) through either PE culture, PE Xpert, or pleural biopsy. RESULTS: A total of 1412 participants underwent screening, and 1344 (95.2%) were subsequently enrolled in this study. Data from 1241 (92.3%) participants were included, comprising 284 with definite TP, 677 with clinically diagnosed TP, and 280 without TP. The sensitivity of cf-TB testing in definite TP was 73.6% (95% CI 68.2-78.4), significantly higher than both Xpert (40.8%, 95% CI 35.3-46.7, P < 0.001) and mycobacterial culture (54.2%, 95% CI 48.4-59.9, P < 0.001). When clinically diagnosed TP was incorporated into the composite reference standard for sensitivity analysis, cf-TB testing showed a sensitivity of 46.8% (450/961, 95% CI 43.7-50.0), significantly higher than both Xpert (116/961, 12.1%, 95% CI 10.2-14.3, P < 0.001) and mycobacterial culture (154/961, 16.0%, 95% CI 13.8-18.5, P < 0.001). The specificities of cf-TB testing, Xpert, and mycobacterial culture were all 100.0%. CONCLUSIONS: The performance of cf-TB testing is significantly superior to that of Xpert and mycobacterial culture methods, indicating that it can be considered as the primary diagnostic approach for improving TP detection. Trial registration The trial was registered on Chictr.org.cn (ChiCTR2000031680, https://www.chictr.org.cn/showproj.html?proj=49316 ).

Bacillus paralicheniformis, an acetate-producing probiotic, alleviates ulcerative colitis via protecting the intestinal barrier and regulating the NLRP3 inflammasome.

Ulcerative colitis (UC) presents a challenging scenario in digestive health, characterized by recurrent inflammation that is often hard to manage. Bacteria capable of producing short-chain fatty acids (SCFAs) play a pivotal role in mitigating UC symptoms, rendering them promising candidates for probiotic therapy. In this investigation, we assessed the impact of Bacillus paralicheniformis HMPM220325 on dextran sodium sulfate (DSS)-induced UC in mice. Genomic analysis of the strain revealed the presence of protease genes associated with acetate and butyrate synthesis, with acetic acid detected in its fermentation broth. Administration of B. paralicheniformis HMPM220325 to UC mice ameliorated pathological manifestations of the condition and restored intestinal barrier function. Furthermore, B. paralicheniformis HMPM220325 suppressed the activation of the NLRP3 inflammasome signaling pathway and modulated the composition of the intestinal microbiota. These findings shed significant light on the potential of B. paralicheniformis as a probiotic candidate, offering a novel avenue for the prevention and therapeutic intervention of colitis.

Curcumin analogue AC17-loaded dissolvable microneedles activate FOXO3 and enhance localized drug delivery for oral squamous cell carcinoma treatment.

Curcumin, a polyphenol extracted from turmeric, is a potential alternative for the treatment of oral squamous cell carcinoma (OSCC) due to its remarkable anticancer activity and low systemic toxicity. To further enhance the anticancer activity and bioavailability of curcumin, we synthesized a curcumin analogue, AC17, by modifying the benzene ring and methylene group of curcumin. A soluble hyaluronic acid microneedle patch (AC17@HAMN) was developed to ensure accurate and safe delivery of AC17 to tumor tissues. The inhibitory effect of AC17 on OSCC cells was stronger than that of curcumin and some common analogues. Transcriptome sequencing showed that the target genes of AC17 were mainly concentrated in apoptosis, cell cycle and cell senescence pathways. Among them, AC17 induces cell cycle arrest and inhibits cell proliferation mainly by activating FOXO3 signaling. With good penetration and dissolution properties, microneedles can deliver AC17 directly to the tumor site and show good anti-tumor effect. Moreover, AC17@HAMN showed good biosafety. In summary, AC17@HAMN offers high efficiency, minimal invasiveness, and few adverse reactions. This microneedle patch holds great promise for potential clinical applications, especially for the treatment of OSCC.

Preparation of waterborne anti-counterfeiting ink based on dual luminescent nanohybrids of bacterial cellulose nanocrystals and lanthanide­nitrogen co-modified GQDs.

To address the growing challenge of counterfeit prevention, this study developed a novel anti-counterfeiting ink system based on bacterial cellulose nanocrystals (BCNC) and lanthanide (Er, Yb)­nitrogen (N) co-dropped graphene quantum dots (GQDs), which exhibited both photoluminescence (PL) and upconversion photoluminescence (UCPL) fluorescent properties as well as excellent rheological characteristics. The Er/Yb/N-GQDs with positive charges were synthesized by a one-step hydrothermal method and subsequently assembled with negatively charged BCNC through electrostatic self-assembly to fabricate a novel nanohybrid, Er/Yb/N-GQDs-BCNC. Raman spectroscopy results indicated an enhancement in the graphitization of GQDs due to lanthanide modification. The TEM results demonstrated a homogeneous distribution of Er/Yb/N-GQDs on BCNC, while XRD, FTIR, and XPS analyses confirmed their physical binding, thus validating the successful synthesis of novel nanohybrids. Then, Er/Yb/N-GQDs-BCNC was introduced into PVA waterborne ink and exhibited dual anti-counterfeiting properties by emitting blue fluorescence at Em 440 nm under Ex 370 nm and green fluorescence at Em 550 nm under Ex 980 nm. Furthermore, the incorporation of BCNC significantly enhanced the thixotropic behavior and yield stress of the PVA waterborne ink. This enhancement made the dual anti-counterfeiting fluorescent ink more suitable for diversified applications on different devices and various substrates, thus providing a novel approach for convenient and rapid information encryption and high security anti-counterfeiting.

Evaluation of droplet digital polymerase chain reaction by detecting cell-free deoxyribonucleic acid in pleural effusion for the diagnosis of tuberculous pleurisy: a multicentre cohort study.

OBJECTIVES: Tuberculous pleurisy is one of the most common types of extra-pulmonary tuberculosis, but the sensitivity of conventional mycobacterial culture (Culture) or Xpert MTB/RIF assay (Xpert) is not satisfying. This multicentre cohort study evaluated the accuracy of a new cell-free DNA droplet digital PCR assay (cf-ddPCR) for diagnosing tuberculous pleurisy. METHODS: Patients with suspected tuberculosis (≥5 years of age) with pleural effusion were consecutively recruited from nine research sites across six provinces in China between September 2020 to May 2022. Culture, Xpert, Xpert MTB/RIF Ultra assay (Ultra), real-time PCR, and cf-ddPCR were performed simultaneously for all specimens. RESULTS: A total of 321 participants were enrolled, and data from 281 (87.5%) participants were available, including 105 definite tuberculous pleurisy, 113 possible tuberculous pleurisy and 63 non-tuberculous pleurisy according to the composite reference standard. The sensitivity of cf-ddPCR was 90.5% (95/105, 95% CI, 82.8-95.1%) in the definite tuberculous pleurisy group, which was significantly higher than those of Culture (57.1%, 60/105, 95% CI, 47.1-66.6%, p < 0.001), Xpert (46.7%, 49/105, 95% CI, 37.0-56.6%, p < 0.001), Ultra (69.5%, 73/105, 95% CI, 59.7-77.9%, p < 0.001) and real-time PCR (75.2%, 79/105, 95% CI, 65.7-82.9%, p < 0.001). In possible tuberculous pleurisy, whose results of Culture and Xpert were both negative, the sensitivity of cf-ddPCR was 61.1% (69/113, 95% CI, 51.4-70.0%), which was still significantly higher than that of Ultra (27.4%, 31/113, 95% CI, 19.7-36.8%, p < 0.001) and real-time PCR (38.9%, 44/113, 95% CI, 30.0-48.6%, p < 0.001). DISCUSSION: The performance of cf-ddPCR is superior to Culture, Xpert, Ultra, and real-time PCR, indicating that improved diagnostic accuracy can be anticipated by incorporating this new assay.

Selenium maintains intestinal epithelial cells to activate M2 macrophages against deoxynivalenol injury.

Selenium (Se) is indispensable in alleviating various types of intestinal injuries. Here, we thoroughly investigated the protective effect of Se on the regulation of the epithelial cell-M2 macrophages pathway in deoxynivalenol (DON)-induced intestinal damage. In the present study, Se has positive impacts on gut health by improving gut barrier function and reducing the levels of serum DON in vivo. Furthermore, our study revealed that Se supplementation increased the abundances of GPX4, p-PI3K, and AKT, decreased the levels of 4-HNE and inhibited ferroptosis. Moreover, when mice were treated with DON and Fer-1(ferroptosis inhibitor), ferroptosis was suppressed and PI3K/AKT pathway was activated. These results indicated that GPX4-PI3K/AKT-ferroptosis was a predominant pathway in DON-induced intestinal inflammation. Interestingly, we discovered that both the number of M2 anti-inflammatory macrophages and the levels of CSF-1 decreased while the pro-inflammatory cytokine IL-6 increased in the intestine and MODE-K cells supernatant. Therefore, Se supplementation activated the CSF-1-M2 macrophages axis, resulting in a decrease in IL-6 expression and an enhancement of the intestinal anti-inflammatory capacity. This study provides novel insights into how intestinal epithelial cells regulate the CSF-1-M2 macrophage pathway, which is essential in maintaining intestinal homeostasis confer to environmental hazardous stimuli.

Identification of Characteristic Bioactive Compounds in Silkie Chickens, Their Effects on Meat Quality, and Their Gene Regulatory Network.

Silkie chicken, an important chicken breed with high medicinal and nutritional value, has a long history of being used as a dietary supplement in China. However, the compounds with health-promoting effects in Silkie chickens remain unclear. In the present study, we conducted a comprehensive analysis of metabolic and lipidomic profiles to identify the characteristic bioactive compounds in Silkie chickens, using a common chicken breed as control. The results showed that the levels of 13 metabolites including estradiol, four lipid subclasses including cardiolipin (CL), eight lipid molecules, and three fatty acids including docosahexaenoic acid (C22:6) were significantly increased in Silkie chickens, which have physiological activities such as resisting chronic diseases and improving cognition. These characteristic bioactive compounds have effects on meat quality characteristics, including improving its water-holding capacity and umami taste and increasing the content of aromatic compounds and phenols. The differentially expressed genes (DEGs) between the two chicken breeds revealed the regulatory network for these characteristic bioactive compounds. Fifteen DEGs, including HSD17B1, are involved in the synthesis of characteristic metabolites. Eleven DEGs, including ELOVL2, were involved in the synthesis and transport of characteristic lipids and fatty acids. In summary, we identified characteristic bioactive compounds in Silkie chickens, and analyzed their effects on meat quality characteristics. This study provided important insight into Silkie chicken meat as a functional food.

Porphyrin fluorescence imaging for real-time monitoring and visualization of the freshness of beef stored at different temperatures.

This study presents a novel fluorescence imaging method for the real-time monitoring of beef quality deterioration and freshness. The fluorescence property of porphyrin in the form of heme can be used to characterize quality changes in beef during storage. Therefore, a fluorescence imaging system with an excitation light source of 440 nm and a CCD camera with a specific wavelength filter of 595 nm was constructed, and the porphyrin fluorescence images of beef samples stored at different temperatures were then collected. The quantitative model for predicting the microbial freshness indicator (TVC) of beef was built with the support vector machine regression (SVR) algorithm and produced satisfactory results with Rc2 and Rp2 of 0.858 and 0.812, respectively. The classification model based on the support vector machine (SVM) algorithm classified beef freshness into "fresh" and "spoiled", with calibration and prediction accuracy of 100 % and 90.9 %, respectively.

Synchronous fluorescence detection of nitrite in meat products based on dual-emitting dye@MOF and its portable hydrogel test kit.

A novel ratiometric fluorescent nanoprobe (Rh6G@UIO-66-NH2) was fabricated for efficient nitrite (NO2-) detection in the present study. When NO2- was introduced, it interacted with the amino groups on the surface of Rh6G@UIO-66-NH2, forming diazonium salts that led to the quenching of blue fluorescence. With this strategy, a good linear relationship between NO2- concentration and the fluorescent intensity ratio of the nanoprobe in the range of 1-100 µM was established, with a detection limit of 0.021 µM. This dual-readout nanosensor was applied to analyze the concentration of NO2- in real meat samples, achieving satisfactory recovery rates of 94.72-104.52%, highlighting the practical potential of this method. Furthermore, a portable Gel/Rh6G@UIO-66-NH2 hydrogel test kit was constructed for on-spot dual-mode detection of NO2-. This kit allows for convenient colorimetric analysis and fluorometric detection when used in conjunction with a smartphone. All the photos taken with the portable kit was converted into digital information using ImageJ software. It provides colorimetric and fluorescent visual detection of NO2- over a range of 0.1-1.5 mM, achieving a direct quantitative tool for NO2- identification. This methodology presents a promising strategy for NO2- detection and expands the application prospects for on-spot monitoring of food safety assessment.

Applications of Gas Sensing in Food Quality Detection: A Review.

Food products often face the risk of spoilage during processing, storage, and transportation, necessitating the use of rapid and effective technologies for quality assessment. In recent years, gas sensors have gained prominence for their ability to swiftly and sensitively detect gases, making them valuable tools for food quality evaluation. The various gas sensor types, such as metal oxide (MOX), metal oxide semiconductor (MOS) gas sensors, surface acoustic wave (SAW) sensors, colorimetric sensors, and electrochemical sensors, each offer distinct advantages. They hold significant potential for practical applications in food quality monitoring. This review comprehensively covers the progress in gas sensor technology for food quality assessment, outlining their advantages, features, and principles. It also summarizes their applications in detecting volatile gases during the deterioration of aquatic products, meat products, fruit, and vegetables over the past decade. Furthermore, the integration of data analytics and artificial intelligence into gas sensor arrays is discussed, enhancing their adaptability and reliability in diverse food environments and improving food quality assessment efficiency. In conclusion, this paper addresses the multifaceted challenges faced by rapid gas sensor-based food quality detection technologies and suggests potential interdisciplinary solutions and directions.

The role of phosphoprotein associated with glycosphingolipid-enriched microdomains 1 (PAG1) in regulating the progression of oral squamous cell carcinoma.

OBJECTIVE: The aim of this study was to explore the role of the tumor suppressor phosphoprotein associated with glycosphingolipid-enriched microdomains 1 (PAG1) on oral squamous cell carcinoma (OSCC) and its molecular mechanism. DESIGN: Immunohistochemistry detected the expression of PAG1 in normal and tumor tissues. The PAG1 overexpressed OSCC cell lines were constructed by lentivirus transfection. Cell Counting Kit-8 assay (CCK-8), clone formation and flow cytometry evaluated the impact of PAG1 on the proliferation and apoptosis of OSCC cells. RNA sequencing (RNA-seq) detected the changes in intracellular genes, and transmission electron microscope (TEM) was used to compare the number of autophagosomes in OSCC cells between Negative and PAG1 group. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and Western blot were used to determine the expression of signaling pathway-related mRNA and proteins respectively. RESULTS: In contrast to the normal tissues, PAG1 expression was significantly downregulated in tumor tissues. Treatment with lentivirus transfection, the expression of PAG1 in the OSCC cell lines was increase. Notably, transfected with PAG1-overexpressing lentivirus cells inhibited the proliferation of OSCC cells and promoted OSCC cells apoptosis. RNA-seq revealed that PAG1 mainly modulated the mitophagy and autophagy pathway, and many autophagosomes were observed in the PAG1 group using TEM. Mechanistically, we found that PAG1 upregulated the expression of autophagy related factors through inhibiting PI3K/Akt/mTOR signal pathway activation. CONCLUSION: Overexpression of PAG1 inhibited OSCC progression by activating autophagy, its mechanism might be related to inhibition of PI3K/Akt/mTOR signal pathway phosphorylation.

Artificial neural network-based shelf life prediction approach in the food storage process: A review.

The prediction of food shelf life has become a vital tool for distributors and consumers, enabling them to determine storage and optimal edible time, thus avoiding unexpected food waste. Artificial neural network (ANN) have emerged as an effective, fast and accurate method for modeling, simulating and predicting shelf life in food. ANNs are capable of tackling nonlinear, complex and ill-defined problems between the variables without prior knowledge. ANN model exhibited excellent fit performance evidenced by low root mean squared error and high correlation coefficient. The low relative error between actual values and predicted values from the ANN model demonstrates its high accuracy. This paper describes the modeling of ANN in food quality prediction, encompassing commonly used ANN architectures, ANN simulation techniques, and criteria for evaluating ANN model performance. The review focuses on the application of ANN for modeling nonlinear food quality during storage, including dairy, meat, aquatic, fruits, and vegetables products. The future prospects of ANN development mainly focus on optimal models and learning algorithm selection, multiple model fusion, self-learning and self-correcting shelf-life prediction model development, and the potential utilization of deep learning techniques.

ANN-based food shelf life prediction methods are reviewed.This paper discusses application of ANN in the food storage process.BPNN is the mainstream ANN architecture used for the prediction of food quality.ANNs are useful for prediction of outputs with high accuracy.Future trends of ANN in the agri-supply chain are evaluated.

QD-based fluorescent nanosensors: Production methods, optoelectronic properties, and recent food applications.

Food quality and safety are crucial public health concerns with global significance. In recent years, a series of fluorescence detection technologies have been widely used in the detection/monitoring of food quality and safety. Due to the advantages of wide detection range, high sensitivity, convenient and fast detection, and strong specificity, quantum dot (QD)-based fluorescent nanosensors have emerged as preferred candidates for food quality and safety analysis. In this comprehensive review, several common types of QD production methods are introduced, including colloidal synthesis, self-assembly, plasma synthesis, viral assembly, electrochemical assembly, and heavy-metal-free synthesis. The optoelectronic properties of QDs are described in detail at the electronic level, and the effect of food matrices on QDs was summarized. Recent advancements in the field of QD-based fluorescent nanosensors for trace level detection and monitoring of volatile components, heavy metal ions, food additives, pesticide residues, veterinary-drug residues, other chemical components, mycotoxins, foodborne pathogens, humidity, and temperature are also thoroughly summarized. Moreover, we discuss the limitations of the QD-based fluorescent nanosensors and present the challenges and future prospects for developing QD-based fluorescent nanosensors. As shown by numerous publications in the field, QD sensors have the advantages of strong anti-interference ability, convenient and quick operation, good linear response, and wide detection range. However, the reported assays are laboratory-focused and have not been industrialized and commercialized. Promising research needs to examine the potential applications of bionanotechnology in QD-based fluorescent nanosensors, and focus on the development of smart packaging films, labeled test strips, and portable kits-based sensors.

Immunoassay with Novel Paired Antibodies for Detection of Lipoarabinomannan in the Pleural Fluid and Plasma of Patients with Tuberculous Pleurisy.

Tuberculous pleurisy (TP) is one of the most common forms of extrapulmonary tuberculosis, but its diagnosis is challenging. Lipoarabinomannan (LAM) antigen is a biomarker for Mycobacterium tuberculosis (Mtb) infection. LAM detection has potential as an auxiliary diagnostic method for TP. We have successfully generated five rabbit anti-LAM monoclonal antibodies (BJRbL01, BJRbL03, BJRbL20, BJRbL52, and BJRbL76). Here, anti-LAM antibodies were tested to detect LAM in the pleural fluid and plasma of patients with TP by sandwich enzyme-linked immunosorbent assays (ELISAs). The results revealed that all of the anti-LAM antibodies were successfully used as capture and detection antibodies in sandwich ELISAs. The BJRbL01/BJRbL01-Bio pair showed better performance than the other antibody pairs for detecting mycobacterial clinical isolates and had a limit of detection of 62.5 pg/mL for purified LAM. LAM levels were significantly higher in the pleural fluid and plasma of patients with TP than in those of patients with malignant pleural effusion or the plasma of non-TB, and LAM levels in the pleural fluid and plasma were positively correlated. Moreover, LAM levels in the pleural fluid sample were significantly higher in confirmed TP patients than in clinically diagnosed TP patients. Our studies provide novel LAM detection choices in the pleural fluid and plasma of TP patients and indicate that LAM detection assay has an auxiliary diagnostic value for TP, which may help to improve the diagnosis of TP.

The species distribution and antimicrobial resistance profiles of Nocardia species in China: A systematic review and meta-analysis.

BACKGROUND: Nocardia species can cause local or disseminated infection. Prompt diagnosis and appropriate treatment of nocardiosis are required, because it can cause significant morbidity and mortality. Knowledge of local species distribution and susceptibility patterns is important to appropriate empiric therapy. However, knowledge on the epidemiology and antimicrobial susceptibility profiles of clinical Nocardia species remains limited in China. METHODS: The data of isolation of Nocardia species were collected from databases such as Pubmed, Web of Science, Embase as well as Chinese databases (CNKI, Wanfang and VIP). Meta-analysis was performed using RevMan 5.3 software. Random effect models were used and tested with Cochran's Q and I2 statistics taking into account the possibility of heterogeneity between studies. RESULTS: In total, 791 Nocardia isolates were identified to 19 species levels among all the recruited studies. The most common species were N. farcinica (29.1%, 230/791), followed by N. cyriacigeorgica (25.3%, 200/791), N. brasiliensis (11.8%, 93/791) and N. otitidiscaviarum (7.8%, 62/791). N. farcinica and N. cyriacigeorgica were widely distributed, N. brasiliensis mainly prevalent in the south, N. otitidiscaviarum mainly distributed in the eastern coastal provinces of China. Totally, 70.4% (223/317) Nocardia were cultured from respiratory tract specimens, 16.4% (52/317) from extra-pulmonary specimens, and 13.3% (42/317) from disseminated infection. The proportion of susceptible isolates as follows: linezolid 99.5% (197/198), amikacin 96.0% (190/198), trimethoprim-sulfamethoxazole 92.9% (184/198), imipenem 64.7% (128/198). Susceptibility varied by species of Nocardia. CONCLUSIONS: N. farcinica and N. cyriacigeorgica are the most frequently isolated species, which are widely distributed in China. Pulmonary nocardiosis is the most common type of infection. Trimethoprim-sulfamethoxazole can still be the preferred agent for initial Nocardia infection therapy due to the low resistance rate, linezolid and amikacin could be an alternative to treat nocardiosis or a choice in a combination regimen.

Single-cell profiling reveals distinct immune response landscapes in tuberculous pleural effusion and non-TPE.

Background: Tuberculosis (TB) is caused by Mycobacterium tuberculosis (Mtb) and remains a major health threat worldwide. However, a detailed understanding of the immune cells and inflammatory mediators in Mtb-infected tissues is still lacking. Tuberculous pleural effusion (TPE), which is characterized by an influx of immune cells to the pleural space, is thus a suitable platform for dissecting complex tissue responses to Mtb infection. Methods: We employed singe-cell RNA sequencing to 10 pleural fluid (PF) samples from 6 patients with TPE and 4 non-TPEs including 2 samples from patients with TSPE (transudative pleural effusion) and 2 samples with MPE (malignant pleural effusion). Result: Compared to TSPE and MPE, TPE displayed obvious difference in the abundance of major cell types (e.g., NK, CD4+T, Macrophages), which showed notable associations with disease type. Further analyses revealed that the CD4 lymphocyte population in TPE favored a Th1 and Th17 response. Tumor necrosis factors (TNF)-, and XIAP related factor 1 (XAF1)-pathways induced T cell apoptosis in patients with TPE. Immune exhaustion in NK cells was an important feature in TPE. Myeloid cells in TPE displayed stronger functional capacity for phagocytosis, antigen presentation and IFN-γ response, than TSPE and MPE. Systemic elevation of inflammatory response genes and pro-inflammatory cytokines were mainly driven by macrophages in patients with TPE. Conclusion: We provide a tissue immune landscape of PF immune cells, and revealed a distinct local immune response in TPE and non-TPE (TSPE and MPE). These findings will improve our understanding of local TB immunopathogenesis and provide potential targets for TB therapy.

The crescent-like Golgi ribbon is shaped by the Ajuba/PRMT5/Aurora-A complex-modified HURP.

BACKGROUND: Golgi apparatus (GA) is assembled as a crescent-like ribbon in mammalian cells under immunofluorescence microscope without knowing the shaping mechanisms. It is estimated that roughly 1/5 of the genes encoding kinases or phosphatases in human genome participate in the assembly of Golgi ribbon, reflecting protein modifications play major roles in building Golgi ribbon. METHODS: To explore how Golgi ribbon is shaped as a crescent-like structure under the guidance of protein modifications, we identified a protein complex containing the scaffold proteins Ajuba, two known GA regulators including the protein kinase Aurora-A and the protein arginine methyltransferase PRMT5, and the common substrate of Aurora-A and PRMT5, HURP. Mutual modifications and activation of PRMT5 and Aurora-A in the complex leads to methylation and in turn phosphorylation of HURP, thereby producing HURP p725. The HURP p725 localizes to GA vicinity and its distribution pattern looks like GA morphology. Correlation study of the HURP p725 statuses and GA structure, site-directed mutagenesis and knockdown-rescue experiments were employed to identify the modified HURP as a key regulator assembling GA as a crescent ribbon. RESULTS: The cells containing no or extended distribution of HURP p725 have dispersed GA membranes or longer GA. Knockdown of HURP fragmentized GA and HURP wild type could, while its phosphorylation deficiency mutant 725A could not, restore crescent Golgi ribbon in HURP depleted cells, collectively indicating a crescent GA-constructing activity of HURP p725. HURP p725 is transported, by GA membrane-associated ARF1, Dynein and its cargo adaptor Golgin-160, to cell center where HURP p725 forms crescent fibers, binds and stabilizes Golgi assembly factors (GAFs) including TRIP11, GRASP65 and GM130, thereby dictating the formation of crescent Golgi ribbon at nuclear periphery. CONCLUSIONS: The Ajuba/PRMT5/Aurora-A complex integrates the signals of protein methylation and phosphorylation to HURP, and the HURP p725 organizes GA by stabilizing and recruiting GAFs to its crescent-like structure, therefore shaping GA as a crescent ribbon. Therefore, the HURP p725 fiber serves a template to construct GA according to its shape. Video Abstract.

Systemic immune dysregulation in severe tuberculosis patients revealed by a single-cell transcriptome atlas.

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb) infection, is currently the deadliest infectious disease in human that can evolve to severe forms. A comprehensive immune landscape for Mtb infection is critical for achieving TB cure, especially for severe TB patients. We performed single-cell RNA transcriptome and T-cell/B-cell receptor (TCR/BCR) sequencing of 213,358 cells from 27 samples, including 6 healthy donors and 21 active TB patients with varying severity (6 mild, 6 moderate and 9 severe cases). Two published profiles of latent TB infection were integrated for the analysis. We observed an obviously elevated proportion of inflammatory immune cells (e.g., monocytes), as well as a markedly decreased abundance of various lymphocytes (e.g., NK and γδT cells) in severe patients, revealing that lymphopenia might be a prominent feature of severe disease. Further analyses indicated that significant activation of cell apoptosis pathways, including perforin/granzyme-, TNF-, FAS- and XAF1-induced apoptosis, as well as cell migration pathways might confer this reduction. The immune landscape in severe patients was characterized by widespread immune exhaustion in Th1, CD8+T and NK cells as well as high cytotoxic state in CD8+T and NK cells. We also discovered that myeloid cells in severe TB patients may involve in the immune paralysis. Systemic upregulation of S100A12 and TNFSF13B, mainly by monocytes in the peripheral blood, may contribute to the inflammatory cytokine storms in severe patients. Our data offered a rich resource for understanding of TB immunopathogenesis and designing effective therapeutic strategies for TB, especially for severe patients.