Fabrication of hyaluronic acid-inulin coated Enterococcus faecium for colon-targeted delivery to fight Fusobacterium nucleatum.

The abundance of Fusobacterium nucleatum (F. nucleatum) is highly associated with the development and poor prognosis of colorectal cancer (CRC), which is regarded as a promising target for CRC. However, until now, the novel strategy to clear F. nucleatum in the colon and CRC has not been well proposed. Herein, a probiotic strain Enterococcus faecium (E. faecium, EF47) is verified to secrete various organic acids and bacteriocins to exert superior antimicrobial activity towards F. nucleatum. However, the oral delivery of EF47 is affected by the complex digestive tract environment, so we design the hyaluronic acid-inulin (HA-IN) coated EF47 for colon-targeted delivery to fight F. nucleatum. IN can protect EF47 from the harsh gastrointestinal tract environment and is degraded specifically in the colon, acting as prebiotics to further promote the proliferation of EF47. The exposed HA can also enhance the targeting effect to the tumor area via the interaction with the CD44 receptor on the tumor cells, which is confirmed to increase the adhesive ability in tumor tissues and inhibit the growth of F. nucleatum. Therefore, this colon-targeted delivery system provides a novel platform to realize high-activity and adhesive delivery of probiotics to assist the therapeutic efficiency of CRC.

Probiotics Influence Gut Microbiota and Tumor Immune Microenvironment to Enhance Anti-Tumor Efficacy of Doxorubicin.

Probiotics have been reported to influence the gut microbiota and immune system in various diseases. Now, the potential impacts of probiotics on tumor treatment still need to be investigated. In this study, three strains of probiotics, Bifidobacterium breve BBr60 (BBr60), Pediococcus pentosaceus PP06 (PP06), and Bifidobacterium longum subsp. longum BL21 (BL21) were investigated for their combination with chemotherapeutic drugs doxorubicin (DOX). Our study showed that PP06 and BL21 have good performance in gastric acid, bile salt, and intestinal fluid tolerance, antimicrobial activity to pathogenic Staphylococcus aureus, and adhesion to Caco-2 cells. Besides, the probiotics all exhibited antioxidant effect, especially BL21. In vitro cytotoxicity and in vivo animal studies revealed that probiotics used alone could not directly induce anti-tumor effects, but the combination of PP06/BL21 and DOX exhibits a higher inhibition rate than DOX alone, via recruitment and infiltration of immune cells in the tumor region. After 16S rRNA analysis of fecal samples from animal models, it was found that BL21 could increase the abundance of Akkermansia, which may also play a role in regulating the tumor microenvironment to improve immune response. In conclusion, BL21 and PP06 in this study could enhance the anti-tumor efficacy by influencing the gut microbiota and tumor immune microenvironment.

Perilipin 5 protects the mitochondrial oxidative functions and improves the alcoholic liver injury in mice.

BACKGROUND AND AIMS: Alcohol consumption is a well-established risk factor for the onset and progression of hepatic steatosis. Perilipin 5 (Plin5), a lipid droplet protein, is an important protective factor against hepatic lipotoxicity induced by excessive lipolysis, but its role and molecular mechanism in alcoholic liver disease (ALD) are not fully elucidated. METHODS: The optimized National Institute on Alcohol Abuse and Alcoholism model was used to construct ALD model mice. Automatic biochemical analyser was used for Biochemical Parameters. The primary hepatocytes and Plin5-overexpressed HepG2 cells (including full-length Plin5 and Plin5 deleting 444-464 aa) were used for in vitro experiment. Haematoxylin and Eosin staining, Oil Red O staining, Bodipy 493/503 staining, Periodic Acid-Schiff staining, immunohistochemistry and JC-1 staining were used to evaluate cell morphology, lipids, glycogen, inflammation and membrane potential. Commercially kits are used to detect glycolipid metabolites, such as triglycerides, glycogen, glucose, reactive oxygen species, lactic acids, ketone bodies. Fluorescently labelled deoxyglucose, NBDG, was used for glucose intake. An XF96 extracellular flux analyser was used to determinate oxygen consumption rate in hepatocytes. The morphological and structural damage of mitochondria was evaluated by electron microscopy. Classical ultracentrifugation is used to separate the subcellular organelles of tissues and cells. Immunoblotting and qPCR were used to detect changes in mRNA and protein levels of related genes. RESULTS: Our results showed that the expression of Plin5 in mouse livers was enhanced by alcohol intake, and Plin5 deficiency aggravated the alcohol-induced liver injury. To clarify the mechanism, we found that Plin5 deficiency significantly elevated the hepatic NADH levels and ketone body production in the alcohol-treated mice. As NADH elevation could promote the reduction of pyruvate into lactate and then inhibit the gluconeogenesis, alcohol-treated Plin5-deficient mice exhibited more lactate production and severer hypoglycemia. These results implied that Plin5 deficiency impaired the mitochondrial oxidative functions in the presence of alcohol. In addition, we demonstrated that Plin5 could be recruited onto mitochondria by alcohol, while Plin5 without mitochondrial targeting sequences lost its mitochondrial protection functions. CONCLUSION: Collectively, this study demonstrated that the mitochondrial Plin5 could protect the alcohol-induced mitochondrial injury, which provides an important new insight on the roles of Plin5 in highly oxidative tissues.

Perilipin 5 deficiency aggravates cardiac hypertrophy by stimulating lactate production in leptin-deficient mice.

BACKGROUND: Perilipin 5 (Plin5) is well known to maintain the stability of intracellular lipid droplets (LDs) and regulate fatty acid metabolism in oxidative tissues. It is highly expressed in the heart, but its roles have yet to be fully elucidated. METHODS: Plin5-deficient mice and Plin5/leptin-double-knockout mice were produced, and their histological structures and myocardial functions were observed. Critical proteins related to fatty acid and glucose metabolism were measured in heart tissues, neonatal mouse cardiomyocytes and Plin5-overexpressing H9C2 cells. 2-NBDG was employed to detect glucose uptake. The mitochondria and lipid contents were observed by MitoTracker and BODIPY 493/503 staining in neonatal mouse cardiomyocytes. RESULTS: Plin5 deficiency impaired glucose utilization and caused insulin resistance in mouse cardiomyocytes, particularly in the presence of fatty acids (FAs). Additionally, Plin5 deficiency increased the NADH content and elevated the expression of lactate dehydrogenase (LDHA) in cardiomyocytes, which resulted in increased lactate production. Moreover, when fatty acid oxidation was blocked by etomoxir or LDHA was inhibited by GSK2837808A in Plin5-deficient cardiomyocytes, glucose utilization was improved. Leptin-deficient mice exhibited myocardial hypertrophy, insulin resistance and altered substrate utilization, and Plin5 deficiency exacerbated myocardial hypertrophy in leptin-deficient mice. CONCLUSION: Our results demonstrated that Plin5 plays a critical role in coordinating fatty acid and glucose oxidation in cardiomyocytes, providing a potential target for the treatment of metabolic disorders in the heart.

Clinical data analysis of 86 patients with invasive infections caused by Malassezia furfur from a tertiary medical center and 37 studies.

Objective: Malassezia furfur (M. furfur) is a lipophilic, conditionally pathogenic yeast that mainly causes skin infections, but the reports of related invasive infections are increasing. The aim of this study is to provide clinical data to assist physicians in the management of patients with invasive infections caused by M. furfur. Methods: A case of pulmonary infection caused by M. furfur in a hematopoietic stem cell transplant patient for aplastic anemia was reported. In addition, the literature on invasive infection by M. furfur published in PubMed and Web of Science in English until 31 July 2022 was reviewed. Results: Clinical data analysis of 86 patients (from 37 studies and our case) revealed that most of them were preterm (44.2%), followed by adults (31.4%). M. furfur fungemia occurred in 79.1% of the 86 patients, and 45 of them were clearly obtained from catheter blood. Other patients developed catheter-related infections, pneumonia, peripheral thromboembolism, endocarditis, meningitis, peritonitis and disseminated infections. Thirty-eight preterm infants had underlying diseases such as very low birth weight and/or multiple organ hypoplasia. The remaining patients had compromised immunity or severe gastrointestinal diseases. 97.7% of patients underwent invasive procedures and 80.2% received total parenteral nutrition (TPN). Fever, thrombocytopenia and leukocytosis accounted for 55.8%, 38.4% and 24.4% of patients with M. furfur invasive infections, respectively. 69.8% of the patients received antifungal therapy, mainly amphotericin B (AmB) or azoles. Of 84 patients with indwelling catheters, 58.3% underwent the removal of catheters. TPN were discontinued in 30 of 69 patients. The all-cause mortality of 86 patients was 27.9%. Conclusions: M. furfur can cause a variety of invasive infections. These patients mostly occur in premature infants, low immunity and severe gastrointestinal diseases. Indwelling catheters and TPN infusion are major risk factors. AmB, l-AmB and azoles are the most commonly used agents, and simultaneous removal of the catheter and termination of TPN infusion are important for the treatment of M. furfur invasive infections.

Prediction of prokaryotic and eukaryotic promoters based on information-theoretic features.

Promoters are DNA regulatory elements located near the transcription start site and are responsible for regulating the transcription of genes. DNA fragments arranged in a certain order form specific functional regions with different information contents. Information theory is the science that studies the extraction, measurement and transmission of information. The genetic information contained in DNA follows the general laws of information storage. Therefore, method in information theory can be used for the analysis of promoters carrying genetic information. In this study, we introduced the concept of information theory to the study of promoter prediction. We used 107 features extracted based on information theory methods and a backpropagation neural network to build a classifier. Then, the trained classifier was applied to predict the promoters of 6 organisms. The average AUCs of the 6 organisms obtained by using hold-out validation and ten-fold cross-validation were 0.885 and 0.886, respectively. The results verified the effectiveness of information-theoretic features in promoter prediction. Considering the possible redundancy in the feature set, we performed feature selection and obtained key feature subsets related to promoter characteristics. The results indicate the potential utility of information-theoretic features in promoter prediction.

Deafness gene screening based on a multilevel cascaded BPNN model.

Sudden sensorineural hearing loss is a common and frequently occurring condition in otolaryngology. Existing studies have shown that sudden sensorineural hearing loss is closely associated with mutations in genes for inherited deafness. To identify these genes associated with deafness, researchers have mostly used biological experiments, which are accurate but time-consuming and laborious. In this paper, we proposed a computational method based on machine learning to predict deafness-associated genes. The model is based on several basic backpropagation neural networks (BPNNs), which were cascaded as multiple-level BPNN models. The cascaded BPNN model showed a stronger ability for screening deafness-associated genes than the conventional BPNN. A total of 211 of 214 deafness-associated genes from the deafness variant database (DVD v9.0) were used as positive data, and 2110 genes extracted from chromosomes were used as negative data to train our model. The test achieved a mean AUC higher than 0.98. Furthermore, to illustrate the predictive performance of the model for suspected deafness-associated genes, we analyzed the remaining 17,711 genes in the human genome and screened the 20 genes with the highest scores as highly suspected deafness-associated genes. Among these 20 predicted genes, three genes were mentioned as deafness-associated genes in the literature. The analysis showed that our approach has the potential to screen out highly suspected deafness-associated genes from a large number of genes, and our predictions could be valuable for future research and discovery of deafness-associated genes.

Single nucleotide mutation changes the capability of CCN3 in osteosarcoma cell invasion and differentiation.

This study aimed to identify significant mutations in CCN3 gene in osteosarcoma, and to explore the influence of this gene on cell invasion and differentiation and the underlying mechanism. Sanger sequencing was used to identify CCN3 gene sequence in human osteosarcoma cell lines, peripheral blood mononuclear cells (PBMC), and osteosarcoma tissues. Wild-type and mutant CCN3 (mCCN3) were ectopically expressed by lentivirus in human osteosarcoma cell lines. Tumor cell invasion was measured by trans-well assay. Osteogenic differentiation was induced by osteogenic differentiating medium and evaluated based on alkaline phosphatase activity and collagen type I alpha 1 chain and osteocalcin expression. Western blotting was used to detect protein levels of CCN3 and mCCN3 in cytoplasmic, nuclear and secreted fractions of cells. A G-to-A single nucleotide mutation in the coding region of CCN3 was found in both osteosarcoma cells and tissues. The frequency of this mutation in osteosarcoma tissue was much higher than that in para-carcinoma tissue and PBMC of healthy people. This nucleotide mutation decreased nuclear glycosylated full length protein level of CCN3 and affected osteosarcoma cell invasion and differentiation. A lower nuclear ratio of glycosylated/non-glycosylated isoforms accounted for the different behavior of mCCN3 compared with CCN3. The G-to-A mutation identified in CCN3 resulted in differential glycosylated full-length protein levels and altered the functional role of CCN3 in osteosarcoma cell invasion and differentiation.

Clinical Characteristics and Risk Factors for in-Hospital Mortality in 240 Cases of Infective Endocarditis in a Tertiary Hospital in China: A Retrospective Study.

Purpose: This study aimed (i) to investigate the clinical characteristics and risk factors related to in-hospital mortality in patients with infective endocarditis (IE) and (ii) to compare the differences in three age groups. Methods: A total of 240 IE cases diagnosed using the modified Duke criteria between January 2016 and December 2019 were included and retrospectively studied. Patients were stratified into three age groups: < 50 y, 50-65 y, and > 65 y. Results: The mean age of the patients was 51 ± 14 y, and 154 patients (64.2%) were male. In addition, 136 (56.7%) patients with IE had no previous cardiac disease. Congenital heart disease (CHD, 21.3%) was the most common underlying heart disease, followed by rheumatic heart disease (RHD, 8.8%). Streptococcus was found in 55 (22.9%) patients and was the most common causative pathogen, comprising 52.9% of all positive blood cultures. Echocardiography showed the presence of vegetations in 88.3% of cases and the predominant involvement of the left heart valves. Fever and cardiac murmur were the most frequent presentations, with no significant differences among age groups. Compared with younger patients, elderly patients had a lower operation rate and higher in-hospital mortality. The independent risk factors of in-hospital mortality were age > 65 y, intracranial infection, splenic embolization, cerebral hemorrhage, NYHA class III-IV, and prosthetic valve infection. Conclusion: CHD replaces RHD as the most common underlying heart disease in IE patients. Patients without previous cardiac disease are at increased risk of IE. Streptococcus is still the primary causative pathogen of IE. Elderly patients present with more comorbidities and complications, in addition to a more severe prognosis than younger patients. Age older than 65 y, intracranial infection, splenic embolization, cerebral hemorrhage, NYHA class III-IV, and prosthetic valve infection showed poorer in-hospital outcomes.

Prokaryotic and eukaryotic promoters identification based on residual network transfer learning.

Promoters contribute to research in the context of many diseases, such as coronary heart disease, diabetes and tumors, and one fundamental task is to identify promoters. Deep learning is widely used in the study of promoter sequence recognition. Although deep models have fast and accurate recognition capabilities, they are also limited by their reliance on large amounts of high-quality data. Therefore, we performed transfer learning on a typical deep network based on residual ideas, called a deep residual network (ResNet), to solve the problem of a deep network's high dependence on large amounts of data in the process of promoter prediction. We used binary one-hot encoding to represent the promoter and took advantage of ResNet to extract feature representations from organisms with a large amount of promoter data. Then, we transferred the learned structural parameters to target organisms with insufficient promoter data to improve the generalization performance of ResNet in target organisms. We evaluated the promoter datasets of four organisms (Bacillus subtilis, Escherichia coli, Saccharomyces cerevisiae and Drosophila melanogaster). The experimental results showed that the AUCs of ResNet's promoter prediction after deep transfer were 0.8537 and 0.8633, which increased by 0.1513 and 0.1376 in prokaryotes and eukaryotes, respectively.

Septic Shock, Renal Abscess, and Bacteremia Due to Peptoniphilus asaccharolyticus in a Woman with Nephrosis and Diabetes Mellitus: Case Report and Literature Review.

Peptoniphilus asaccharolyticus is a Gram-positive anaerobic coccus, which forms part of the normal flora and the human commensals of the skin, genitourinary system, and gut. It can cause opportunistic infections in immunocompromised patients and is frequently isolated as part of polymicrobial spectra. Severe monomicrobial infections caused by the genus rarely occur. In this study, we report on septic shock, renal abscess, and bacteremia due to P. asaccharolyticus in a woman with nephrosis and diabetes mellitus. To the best of our knowledge, this report is the first to describe P. asaccharolyticus isolated from both renal abscess and blood cultures purely. The underlying diseases of the host and the removal of the double J tube were significant predisposing factors in this infection.

Effect of drying methods on Lactobacillus Rhamnosus GG microcapsules prepared using the complex coacervation method.

Lactobacillus rhamnosus GG (LGG) microcapsules prepared using the complex coacervation method were dried through spray and vacuum freeze drying. Physical properties, resistance to simulated gastrointestinal and bile salt conditions, and the stability of the LGG microcapsules during storage and heat treatment were estimated. Spray and vacuum freeze drying are suitable for LGG microcapsule preparation because of lower than 2.80% of the water content, and higher than 93.21% of the encapsulation efficiency. Under the simulated gastrointestinal condition, the survival rates of two kinds of microencapsulated LGG were over 96%. At 0.6% bile salt concentration, the survival rate of microencapsulated LGG subjected to spray drying was 110.89% higher than that of LGG subjected to vacuum freeze drying. In addition, the stability of the former was better than that of the latter during storage under low water activity condition (0.11). However, under high water activity condition (0.75), the two showed opposite results. The two kinds of microencapsulated LGG were completely inactivated after 4 weeks in 0.75 of water activity. The survival counts of microencapsulated LGG had no significant difference at -18°C after 8 weeks. After 8 weeks at 25°C and 4°C, the survival loss of microencapsulated LGG through drying and vacuum freeze drying was approximately 2 and 1 log CFU/g, respectively. PRACTICAL APPLICATION: LGG is a kind of microorganism beneficial to the human body, but it is easy to inactivate and lose its physiological value. LGG microcapsules can improve the stability of LGG, prolong the release time, and expand its application range. The drying method will affect the properties of LGG microcapsules. So, the characteristics analysis of LGG microcapsules with different drying methods can provide a reference for the preparation and application of LGG microcapsules.

A Rapid Carbapenemase Genes Detection Method with Xpert Carba-R from Positive Blood Cultures Compared with NG-Test Carba 5 and Sequencing.

Objective: The objective of the current study was to evaluate the performance of Xpert Carba-R for the direct detection and identification of carbapenemase genes from positive blood cultures. Methods: Pathogens which extracted from positive blood cultures and identified using MALDI-TOF MS as Enterobacterales were included in this study. Xpert Carba-R was used for the rapid detection of carbapenemase genes from positive blood cultures. NG-Test CARBA 5 and polymerase-chain reaction (PCR) sequencing were used for the detection of carbapenemases and carbapenemase genes in positive blood culture isolates, respectively. Finally, antibiotic susceptibility tests were conducted using the VITEK-2 Compact system. Results: A total of 133 positive blood cultures of Enterobacterales were collected and 27 of them were detected to carry carbapenemase genes using Xpert Carba-R. In comparison with PCR sequencing results, the sensitivity and specificity of Xpert Carba-R and NG-Test CARBA 5 were calculated as 100%. Additionally, Xpert Carba-R could significantly shorten the turnaround time by directly detecting positive blood cultures comparing with NG-Test CARBA 5. For 27 carbapenem-producing strains, the resistance rates of carbapenems and aztreonam were 96.3% and 92.6%, respectively. Strains carrying the blaKPC gene were all sensitive to ceftazidime-avibactam. All strains were sensitive to tigecycline and colistin. Conclusion: Xpert Carba-R is suitable for the rapid detection of main carbapenemase genes from positive blood cultures with high sensitivity and specificity. In comparison with NG-Test CARBA 5 and PCR sequencing methods, the timely and convenient method can be a useful test to guide optimal therapy and infection control.

Kernel Causality Among Teacher Self-Efficacy, Job Satisfaction, School Climate, and Workplace Well-Being and Stress in TALIS.

Teachers play an important role in the educational system. Teacher self-efficacy, job satisfaction, school climate, and workplace well-being and stress are four individual characteristics shown to be associated with tendency to turnover. In this article, data from the Teaching and Learning International Survey (TALIS) 2018 teacher questionnaire are analyzed, with the goal to understand the interplay amongst these four individual characteristics. The main purposes of this study are to (1) measure extreme response style for each scale using unidimensional nominal response models, and (2) investigate the kernel causal paths among teacher self-efficacy, job satisfaction, school climate, and workplace well-being and stress in the TALIS-PISA linked countries/economies. Our findings support the existence of extreme response style, the rational non-normal distribution assumption of latent traits, and the feasibility of kernel causal inference in the educational sector. Results of the present study inform the development of future correlational research and policy making in education.

Predicting essential genes of 37 prokaryotes by combining information-theoretic features.

Essential genes are required for the reproduction and survival of an organism. Rapid identification of essential genes has practical application value in biomedicine. Information theory is a discipline that studies information transmission. Based on the similarity between heredity and information transmission, measures derived from information theory can be applied to genetic sequence analysis on different scales. In this study, we employed 114 features extracted by information theory methods to construct an essential gene prediction model. We applied a backpropagation neural network to construct a classifier and employed it to predict essential genes of 37 prokaryotes. The performance of the classifier was evaluated by applying intra-organism prediction and leave-one-species-out prediction. Among 37 prokaryotes, intra-organism prediction and leave-one-species-out prediction yielded average AUC scores of 0.791 and 0.717, respectively. Considering the potential redundancy in the feature set, we performed feature selection and constructed a key feature subset. In the above two prediction methods, the average AUC scores of 37 organisms obtained by using key features were 0.786 and 0.714, respectively. The results show the potential and universality of information-theoretic features in the study of prokaryotic essential gene prediction.

Histone H3 methyltransferase Ezh2 promotes white adipocytes but inhibits brown and beige adipocyte differentiation in mice.

Obesity is a disease characterized by imbalance between energy intake and expenditure, excessive energy store in white adipocytes, but brown and beige adipocytes consume energy to relieve obesity. In this study, we want to explore the role of the histone H3 methyltransferase Ezh2 in the differentiation of white, brown and beige adipocytes with Ezh2 conditional knockout mice (Ezh2flox/floxPrx1-cre) and mouse embryonic fibroblasts (MEFs). The results showed that Ezh2-deficient mice have a leaner phenotype and less white adipose tissues. The morphological changes in the adipose tissue included smaller white adipose tissue depots, white adipocytes with smaller diameter, smaller lipid droplets inside the brown adipocytes and more beige adipocytes in the Ezh2-deficient mice compared with the control. The differentiation markers of white adipocytes in Ezh2 knockout mice decreased; Ucp1 and other browning markers increased in brown and beige adipocytes. The Ezh2 knockout mice could better tolerate cold stimulation, and they can also resist obesity and insulin resistance induced by a high-fat diet. The Ezh2 inhibitor GSK126 could inhibit the differentiation of MEFs into white adipocytes but promote their differentiation into brown/beige adipocytes. The H3K27me3 demethylase Jmjd3/UTX inhibitor GSKJ4 inhibited MEFs' differentiation into brown/beige adipocytes. These results showed that Ezh2 promotes the differentiation of white adipocytes and inhibits the differentiation of brown and beige adipocytes in vivo and in vitro through its methylase activity and this may represent new knowledge for obesity therapeutic strategy.

A 2-step algorithm combining glutamate dehydrogenase and nucleic acid amplification tests for the detection of Clostridioides difficile in stool specimens.

The optimized diagnosis algorithm of Clostridioides difficile infection (CDI) is worldwide concerns. The purpose of this study was to assess the toxigenic C. difficile test performance and propose an optimal laboratory workflow for the diagnosis of CDI in mild virulent epidemic areas. Diarrhea samples collected from patients were analyzed by glutamate dehydrogenase (GDH), toxin AB (CDAB), and nucleic acid amplification test (NAAT). We assessed the performance of GDH, the GDH-CDAB algorithm, and the GDH-NAAT algorithm using toxigenic culture (TC) as a reference method. In this study, 186 diarrhea samples were collected. The numbers of TC-positive and TC-negative samples were 39 and 147, respectively. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and kappa of the GDH assay were 100%, 80.3%, 57.4%, 100%, and 0.63; of the GDH-CDAB algorithm were 48.7%, 97.3%, 82.6%, 87.7%, and 0.54; and of the GDH-NAAT algorithm were 74.4%, 100%, 100%, 93.6%, and 0.82, respectively. The GDH-NAAT algorithm has great concordance with TC in detecting toxigenic C. difficile (kappa = 0.82), while the sensitivity of the GDH-CDAB algorithm was too low to meet the demand of CDI diagnosis clinically. GDH-NAAT algorithm is recommended for the detection of toxigenic C. difficile with high specificity, increased sensitivity, and cost-effective.

NDRG2 regulates adherens junction integrity to restrict colitis and tumourigenesis.

BACKGROUND: Paracellular barriers play an important role in the pathogenesis of Inflammatory bowel disease (IBD) and maintain gut homeostasis. N-myc downstream-regulated gene 2 (NDRG2) has been reported to be a tumour suppressor gene and to inhibit colorectal cancer metastasis. However, whether NDRG2 affects colitis initiation and colitis-associated colorectal cancer is unclear. METHODS: Intestine-specific Ndrg2 deficiency mice (Ndrg2ΔIEC) were subjected to DSS- or TNBS-induced colitis, and AOM-DSS-induced colitis-associated tumour. HT29 cells, Caco2 cells, primary intestinal epithelial cells (IECs) from Ndrg2ΔIEC mice, mouse embryo fibroblasts (MEFs) from systemic Ndrg2 knockout mice, HEK293 cells and human UC and DC specimens were used to investigate NDRG2 function in colitis and colitis-associated tumour. FINDINGS: Ndrg2 loss led to adherens junction (AJ) structure destruction via E-cadherin expression attenuation, resulting in diminished epithelial barrier function and increased intestinal epithelial permeability. Mechanistically, NDRG2 enhanced the interaction of E3 ligase FBXO11 with Snail, the repressor of E-cadherin, to promote Snail degradation by ubiquitination and maintained E-cadherin expression. In human ulcerative colitis patients, reduced NDRG2 expression is positively correlated with severe inflammation. INTERPRETATION: These findings demonstrate that NDRG2 is an essential colonic epithelial barrier regulator and plays an important role in gut homeostasis maintenance and colitis-associated tumour development. FUNDING: National Natural Science Foundation of China (No. 81770523, 31571437, 81672751), Creative Research Groups of China (No. 81421003), State Key Laboratory of Cancer Biology Project (CBSKL2019ZZ11, CBSKL201406, CBSKL2017Z08 and CBSKL2017Z11), Fund for Distinguished Young Scholars of ShaanXi province (2019JC-22).

False-Negative Serum Cryptococcal Antigen Lateral Flow Immunoassay Result for a Patient with Disseminated Cryptococcal Disease.

Cryptococcus neoformans (C. neoformans) is an opportunistic fungal pathogen to humans, which can be acquired from environmental sources. Its most important virulence factor is its polysaccharide capsule, which can be used for diagnostic tests that identify the cryptococcal antigen (CrAg). The CrAg lateral flow assay (LFA) is a dipstick immunochromatographic assay with high sensitivity and specificity; however, several false-negative cases have been reported. Here, we present a case of a false-negative serum CrAg LFA, in which the blood culture from a matched sample was positive for C. neoformans, thus demonstrating the postzone phenomenon.