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SDF-1α activity is closely related to information transmission and cell migration when contributing to lymphatic metastasis in various tumors. Herein, we explored the interaction among SDF-1α, CXCR4 and PI3K/Akt signaling pathway in gastric cancer (GC) and their roles in this disorder. Human GC cells KATO-III and BMSCs were co-cultured without contact. GC cells were transfected with SDF-1α, CXCR4 inhibitor, and PI3K inhibitor. After examining the efficiency of transfection, cell migration was evaluated using Transwell chamber, and expression SDF-1α, CD133, and CXCR4 was determined by RT-qPCR. With transfection rate of 98%, the number of migrated cells reduced upon inhibition of CXCR4 and PI3K. Luciferase activity in 565 nm are high than CXCR4 inhibition group. (p < 0.05). Likewise, up-regulation of SDF-1α increased the expression of SDF-1 (0.825±0.061), CD133 (0.875±0.058), CXCR4 (0.801±0.052), and Akt (0.852±0.062), compared to the blank group, CXCR4 inhibition group and PI3K inhibition group (p < 0.05). Down-regulation of CXCR4 and PI3K, however, decreased the expression insignificantly (p > 0.05). Collectively, up-regulation of SDF-1α activates CXCR4 signaling pathway of BMSCs and stimulates its downstream PI3K/Akt signaling pathway and and increases the expression of CD133, thereby promoting malignant behaviors of GC cells.

Latency-associated peptide (LAP) was identified as crucial immune regulator in tumor microenvironment (TME) in recent researches. In this study, we aimed to estimate the predictive value of LAP expression for clinical survival and therapeutic response in muscle-invasive bladder cancer (MIBC). Our study encompassed 140 MIBC patients from Zhongshan Hospital (ZSHS cohort), 401 patients from The Cancer Genome Atlas (TCGA cohort) and 195 patients received PDL1 blockade from IMvigor210 trial. Survival analyses were conducted through Kaplan–Meier curve and Cox regression model. LAP expression and its association with immune contexture were evaluated in ZSHS and TCGA cohort. We found that high intratumoral LAP+ cells infiltration anticipated inferior survival and adjuvant chemotherapy (ACT) response, and was closely related to an immunoevasive contexture with increased M2 macrophages, neutrophils and conspicuously a cluster of highly exhausted CD8+ T cells. The combinational analysis of LAP+ cells and CD8+ T cells infiltration stratified patients into distinct risk groups with implications for therapeutic sensitivity to PDL1 blockade and refinement of molecular classification in MIBC. LAP expression was correlated with patients’ inferior prognosis, ACT-tolerance and an immunoevasive TME with exhausted CD8+ T cell infiltration, suggesting that LAP could serve as a promising therapeutic target in MIBC. Simultaneously, our novel TME classification based on LAP+ cells and CD8+ T cells infiltration and its potential in appraising PDL1 blockade application for MIBC patients deserved further validation.

Hypoxia contributes significantly to the development of chemoresistance of many malignancies including esophageal cancer (EC). Accumulating studies have indicated that long non-coding RNAs play important roles in chemotherapy resistance. Here, we identified a novel lncRNA-EMS/miR-758-3p/WTAP axis that was involved in hypoxia-mediated chemoresistance to cisplatin in human EC. Hypoxia induced the expressions of lncRNA EMS and WTAP, and reduced the expression of miR-758-3p in EC cell line ECA-109. In addition, the expressions of EMS and WTAP were required for the hypoxia-induced drug resistance to cisplatin in EC cells, while overexpression of miR-758-3p reversed such chemoresistance. The targeting relationships between EMS and miR-758-3p, as well as miR-758-3p and WTAP, were verified by luciferase-based reporter assays and multiple quantitative assays after gene overexpression/knockdown. Moreover, we found significant correlations between tumor expressions of these molecules. Notably, higher levels of EMS/WTAP, or lower levels of miR-758-3p in tumors predicted worse survivals of EC patients. Furthermore, in a xenograft mouse model, targeted knockdown of EMS and WTAP in ECA-109 cells markedly attenuated the resistance of tumors to cisplatin treatments. Our study uncovers a critical lncRNA-EMS/miR-758-3p/WTAP axis in regulating hypoxia-mediated drug resistance to cisplatin in EC.

Activation of intestinal macrophages is implicated in the pathogenesis of neonatal necrotizing enterocolitis (NEC), yet its precise mechanisms remain unclear. The purpose of this study is to investigate the role of macrophages and TNF-α via an inflammatory MicroRNA in NEC. Immunofluorescence (IF) staining of CD68, iNOS, and Arg-1 was employed to identify phenotypes of macrophage in the intestines of NEC infants and NEC mice. Expression of TNF-α, c-kit, and miR-222 was evaluated by qRT-PCR, Western blot, and immunochemical staining from the tissue samples. Large number of M1 macrophage infiltration was found in the NEC intestines. Expression of CD68, iNOS, and TNF-α were significantly increased, while c-kit was decreased distinctly in the NEC group. In the early phase of NEC mouse model, inhibition of M1 macrophages reduced the incidence of NEC and intestinal inflammation. We found that TNF-α upregulated the expression of miRNA-222 and inhibited the expression of c-kit. Conversely, such decrease of c-kit expression could be reversed by miR-222 antagonists. Furtherly, dual-luciferase assay confirmed that c-kit can be inhibited by miR-222 directly. Macrophages activation in NEC intestine results in an increased inflammatory response and TNF-α production, accompanied with miR-222 upregulation and c-kit suppression. Modulations of M1 macrophages, TNF-α or miR-222 may be potential therapeutic targets for NEC treatment.

Acute pancreatitis results in major mortality and morbidity. Lipase is a vital biomarker of acute pancreatitis in the early stage. Developing a simple and convenient methodology for monitoring the lipase activity is of significance to the early diagnosis of acute pancreatitis. In this work, we designed a novel ratiometric fluorescent probe (CARA) based on FRET for detecting lipase activity in human serum. The energy donor group coumarin derivative (CA) and the energy acceptor group rhodamine derivative (RA) were linked for the construction of a coumarin–rhodamine platform through an ester bond that is easily cleaved by lipase, breaking the FRET process between CA and RA. CARA shows a high selectivity to lipase over other biological substances and a good linearity between the fluorescence ratio and lipase activity in serum samples. CARA has the potential to be applied in the clinical diagnosis of acute pancreatitis.

Objective: The X-ray and CT features of minimally invasive percutaneous pedicle screw for open reduction and internal fixation and traditional open reduction and internal fixation for thoracolumbar fractures were compared. Methods: Sixty-two patients with thoracolumbar fractures were analyzed with single vertebral body injury without obvious osteoporosis. Patients were randomly divided into two groups. In group A, minimally invasive percutaneous pedicle screws were used for open reduction and fixation. Group B was treated with thoracolumbar vertebral fracture with traditional incision and reduction and internal fixation with posterolateral fusion. Both groups followed standard surgical methods and methods, the same postoperative treatment and functional exercise plan. Results: During a 9-month follow-up, X-ray and computed tomography were used to analyze fracture joint motion injuries. The recovery rate of vertebral height in group A was significantly better than that in group B. At 9 months after operation, the recovery rate of vertebral height in group A increased. The neural function was similar, but the improvement rate in group A was significantly better than that in group B. Conclusions: Minimally invasive percutaneous pedicle screws for open reduction and internal fixation are compared with traditional open reduction and internal fixation for thoracolumbar fractures. Vertebral body height, symptom improvement, and neurological recovery after a lumbar fracture. However, minimally invasive percutaneous pedicle screws for open reduction and internal fixation can immediately increase the compression stability of the anterior spine and reduce complications such as nail and rod rupture due to excessive fixation pressure.

Purpose Esophageal cancer is a highly lethal and broad-spreading malignant tumor worldwide. Exosome-carrying lncRNAs play an essential role in the pathogenesis of various cancers. Results The results revealed that the expression of UCA1 was decreased in esophageal cancer tissues and plasma exosomes. UCA1 was enriched in exosomes, and exosomal UCA1 was a promising biomarker for the diagnosis of esophageal cancer with 86.7% sensitivity and 70.2% specificity. Overexpression of UCA1 played anticancer roles in esophageal cancer cells through inhibiting cell proliferation, invasion and migration, and colony formation. Also, exosomal UCA1 was taken up by esophageal cancer cells and inhibited the progression of esophageal cancer in vitro and tumor growth in vivo. Furthermore, exosomal UCA1 could directly target miRNA-613 in esophageal cancer cells. Conclusions The results suggested that exosomal UCA1 inhibits tumorigenesis and progression of esophageal cancer in vitro and in vivo, and might be a promising biomarker for esophageal cancer. Patient and methods In this study, we determined the expression of UCA1 in esophageal cancer tissues, plasma exosomes of patients with esophageal cancer. We determined the potential of exosomal UCA1 as a biomarker and its effect on the pathogenesis and progression of esophageal cancer in vitro and in vivo.

Breast cancer brain metastasis (BCBM) is a devastating disease. Radiation therapy remains the mainstay for treatment of this disease. Unfortunately, its efficacy is limited by the dose that can be safely applied. One promising approach to overcoming this limitation is to sensitize BCBMs to radiation by inhibiting their ability to repair DNA damage. Here, we report a DNA repair suppressor, leucine-rich repeat-containing protein 31 (LRRC31), that was identified through a genome-wide CRISPR screen. We found that overexpression of LRRC31 suppresses DNA repair and sensitizes BCBMs to radiation. Mechanistically, LRRC31 interacts with Ku70/Ku80 and the ataxia telangiectasia mutated and RAD3-related (ATR) at the protein level, resulting in inhibition of DNA-dependent protein kinase, catalytic subunit (DNA-PKcs) recruitment and activation, and disruption of the MutS homologue 2 (MSH2)-ATR module. We demonstrate that targeted delivery of the LRRC31 gene via nanoparticles improves the survival of tumour-bearing mice after irradiation. Collectively, our study suggests LRRC31 as a major DNA repair suppressor that can be targeted for cancer radiosensitizing therapy.

Egg white peptides are well known for their abundant species and biological properties. The aim of this study was to investigate the effect of egg white peptides on skin wound healing. We hydrolyzed the egg white powder using Alcalase and flavourzyme. Our transwell cell migration assay data showed that EWP2, which is one of the four fractions obtained from Sephadex G-15 isolation, promoted human skin fibroblast HSF cells migration. LC–MS/MS analysis results showed EWP2 were consist of 24 high abundant peptides. We randomly chose and synthesized four of these 24 peptides. Transwell assay data displayed that the egg white peptide KPHAEVVLR (KR-9) increased 8.2-fold of HSF cells migration at 200 μM compared with control. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sμlfophenyl)-2H-tetrazolium (MTS) assay results showed KR-9 did not lead to HSF cells over-proliferation. In vivo animal experiment data proved that KR-9 accelerated 50 % and 136 % mice skin wound healing rate at 50 μM and 100 μM. Furthermore, we explored the mechanism of KR-9. Our western blot and immunofluorescence results showed KR-9 enhanced the protein level of cell membrane heat shock protein 90α (Hsp90α). Thus, we conclude that egg white peptide KR-9 promotes skin fibroblasts migration and mice skin wound healing by stimulating cell membrane Hsp90α secretion.

Introduction The objective of this study is to investigate the role of thyroid hormone (TH) in the pathogenesis of intestinal dysganglionosis (ID). Methods A zebrafish model of congenital hypothyroidism (CH) was created by exposing the larvae to the 6-propyl-2-thiouracil (PTU). The enteric neurons were labeled with anti-HuC/D antibodies. The number of enteric neurons was counted. The larval intestine was dissociated and stained with anti-p75 and anti-α4 integrin antibodies. Mitosis and apoptosis of the p75+ α4 integrin+ enteric neural crest cells (ENCCs) were studied using flow cytometry. Intestinal motility was studied by analyzing the transit of fluorescent tracers. Results PTU (25 mg/L) significantly reduced TH production at 6- and 9-days post fertilization without changing the body length, body weight, and intestinal length of the larvae. Furthermore, PTU inhibited mitosis of ENCCs and reduced the number of enteric neurons throughout the larval zebrafish intestine. Importantly, PTU inhibited intestinal transit of fluorescent tracers. Finally, thyroxine supplementation restored ENCC mitosis, increased the number of enteric neurons, and recovered intestinal motility in the PTU-treated larvae. Conclusions PTU inhibited TH production, reduced the number of enteric neurons, impaired intestinal motility, and impeded ENCC mitosis in zebrafish, suggesting a possible role of CH in the pathogenesis of ID.

Mucin 13 (MUC13) is a glycoprotein that is expressed on the cell surface and participates in the tumorigenesis of multiple malignancies, including pancreatic cancer, colorectal cancer and renal cancer. However, to the best of our knowledge, the expression levels and function of MUC13 in lung cancer progression have not yet been demonstrated. Therefore, the present study examined the expression pattern and regulatory role of MUC13 in lung cancer tumorigenesis. The results demonstrated that MUC13 was highly expressed in lung cancer tissues and cell lines compared with that in normal tissues and cell lines. Functionally, knockdown of MUC13 inhibited cell proliferation and enhanced the apoptosis of A549 and NCI-H1650 lung cancer cells. Furthermore, silencing of MUC13 suppressed the migration and invasion of lung cancer cells. Additionally, a xenograft tumor model demonstrated that knockdown of MUC13 delayed the development of the lung cancer xenograft and suppressed the expression of proliferation marker Ki-67 in tumor tissues. Mechanistically, MUC13 activated the ERK signaling pathway by enhancing the phosphorylation of ERK, JNK and p38 in lung cancer tissues compared with that in normal tissues. Knockdown of MUC13 inhibited the phosphorylation of ERK/JNK/p38 in A549 and NCI-H1650 cells. Overall, these findings suggested that MUC13 could act as an oncogenic glycoprotein to accelerate the progression of lung cancer via abnormal activation of the ERK/JNK/p38 signaling pathway and might serve as a therapeutic target for lung cancer treatment.

Housing systems are among the most important non-genetic factors affecting hen production performance and intestinal microbes. With increased interest in animal welfare, cage-free laying hen housing systems have become common, providing behavioral freedom and health benefits. The present study aimed to compare the effects of plastic net housing system (NRS) and floor litter housing system (LRS) on the composition and function of the duodenal and cecal microbiota in Shendan chicken, one of the most popular laying hen strains in China. The associations between the differential microbiota abundance and production traits and intestinal morphological parameters were determined. Compared with the LRS, the NRS improved the laying rate (p < 0.05) and increased the villus height (VH) of the duodenum (p < 0.05) and the VH-to-crypt depth ratio (VCR) of the cecum (p < 0.05). Alpha diversity analysis showed that LRS chickens had a significantly higher diversity and richness than NRS chickens. Beta diversity analysis demonstrated differences in the microbiota composition based on housing systems. Within the cecum, Proteobacteria and Kiritimatiellaeota were significantly more abundant in the LRS than in the NRS (p < 0.05), while Bacteroidetes were significantly less abundant in the LRS (p < 0.05). Phascolarctobacterium and Ruminococcaceae_UCG-005 were significantly less abundant in the LRS (p < 0.05) compare to the NRS. Within the duodenum, Lactobacillus was significantly less abundant in the LRS (p < 0.05) than in the NRS, while Pseudomonas was significantly more abundant in the LRS (p < 0.05). Cecal Phascolarctobacterium and Ruminococcaceae_UCG-005 were significantly positively correlated with eggshell strength (R = 0.608, p < 0.01) and egg weight (R = 0.526, p < 0.05), respectively. Duodenal Lactobacillus was significantly positively correlated with VH and VCR (R = 0.548 and 0.565, p < 0.05), while Pseudomonas was significantly negatively correlated with the Haugh unit (R = −0.550, p < 0.05). In conclusion, there are differences in the cecal and duodenal microbiota compositions of Shendan laying hens reared in different non-cage housing systems, and the NRS was superior to the LRS in improving the laying performance and intestinal morphology and microecological environment.

This study explored the role played by combined ICA and bone mesenchymal stem cells (BMSCs) in repairing rabbit knee cartilage defects. Firstly, rabbit BMSCs were isolated and used to construct an in vitro cellular model of oxygen‐glucose deprivation/reoxygenation (OGD/R). Subsequently, ICA processing, Alcian blue staining, immunofluorescence and Western blot studies were performed to evaluate the ability of BMSCs to display signs of chondrogenic differentiation. Furthermore, a rabbit knee cartilage injury model was established in vivo. International Cartilage Repair Society (ICRS) macroscopic evaluations, H&E, Alcian blue and EdU staining, as well as immunohistochemistry, were analysed cartilage repair and pathological condition of the knee cartilage tissue. Our in vitro results showed that ICA promoted the chondrogenic differentiation of BMSCs, as well as aggrecan (AGR), bone morphogenetic protein 2 (BMP2) and COL2A1 protein expression in BMSCs. In vivo experiments showed that rabbits in the BMSCs or ICA treatment group had higher ICRS scores and displayed a better restoration of cartilage‐like tissue and chondrocyte expression on the surface of their cartilage defects. In conclusion, ICA or BMSCs alone could repair rabbit knee cartilage damage, and combined treatment with ICA and BMSCs showed a better ability to repair rabbit knee cartilage damage.

Background To investigate the association between CLEC10A and prognosis in breast cancer (BC) patients. Methods We assessed the prognostic value of CLEC10A in BC using data from The Cancer Genome Atlas (TCGA) online database. We examined CLEC10A expression differences in BC and normal tissues via the TIMER and UALCAN databases. Then, we used the Kaplan-Meier plotter database to evaluate the correlation of CLEC10A mRNA levels with clinical outcomes. Subsequently, the TIMER platform and TISIDB website were used to assess the correlation of CLEC10A with the tumor immune cell infiltration level in BC. Results Our results showed that CLEC10A levels were significantly downregulated in BC tissues compared with normal tissues. CLEC10A expression was associated with histologic type, pathologic stage, T stage, Her2 status and a poor prognosis. Additionally, CLEC10A was positively related to the level of different tumor-infiltrating immune cells in BC, and CLEC10A was closely correlated with the gene markers of diverse immune cells. Additionally, low CLEC10A expression predicted a poor prognosis in BC patients grouped based on immune cell infiltration levels. Conclusion CLEC10A may be a potential biomarker and may efficiently predict prognosis in BC patients.

Rapid COVID-19 diagnosis in the hospital is essential, although this is complicated by 30%–50% of nose/throat swabs being negative by SARS-CoV-2 nucleic acid amplification testing (NAAT). Furthermore, the D614G spike mutant dominates the pandemic and it is unclear how serological tests designed to detect anti-spike antibodies perform against this variant. We assess the diagnostic accuracy of combined rapid antibody point of care (POC) and nucleic acid assays for suspected COVID-19 disease due to either wild-type or the D614G spike mutant SARS-CoV-2. The overall detection rate for COVID-19 is 79.2% (95% CI 57.8–92.9) by rapid NAAT alone. The combined point of care antibody test and rapid NAAT is not affected by D614G and results in very high sensitivity for COVID-19 diagnosis with very high specificity., Graphical Abstract, Highlights Combined rapid antibody + nucleic acid detection correctly diagnoses SARS-CoV-2 Rapid antibody tests detect immune responses against SARS-CoV-2 bearing D614G Rapid SARS-CoV-2 antibody tests do not cross-react with antibodies to seasonal CoV False positivity in SARS-CoV-2 finger prick blood antibody tests can be very low, Mlcochova et al. report that combined rapid nucleic acid amplification testing (NAAT) and finger prick blood antibody tests can substantially improve the diagnosis of COVID-19 as compared to NAAT alone and is able to detect the SARS-CoV-2 Spike D614G variant that dominates the pandemic.

Graphical abstract, The main protease (Mpro or 3CLpro) of SARS-CoV-2 virus is a cysteine enzyme critical for viral replication and transcription, thus indicating a potential target for antiviral therapy. A recent repurposing effort has identified ebselen, a multifunctional drug candidate as an inhibitor of Mpro. Our docking of ebselen to the binding pocket of Mpro crystal structure suggests a noncovalent interaction for improvement of potency, antiviral activity and selectivity. To test this hypothesis, we designed and synthesized ebselen derivatives aimed at enhancing their non-covalent bonds within Mpro. The inhibition of Mpro by ebselen derivatives (0.3 μM) was screened in both HPLC and FRET assays. Nine ebselen derivatives (EBs) exhibited stronger inhibitory effect on Mpro with IC50 of 0.07–0.38 μM. Further evaluation of three derivatives showed that EB2-7 exhibited the most potent inhibition of SARS-CoV-2 viral replication with an IC50 value of 4.08 µM in HPAepiC cells, as compared to the prototype ebselen at 24.61 μM. Mechanistically, EB2-7 functions as a noncovalent Mpro inhibitor in LC-MS/MS assay. Taken together, our identification of ebselen derivatives with improved antiviral activity may lead to developmental potential for treatment of COVID-19 and SARS-CoV-2 infection.

Countries’ increases in testing capacity during the first waves of the COVID-19 pandemic, coupled with reductions in case numbers between waves, have resulted in shifts from diagnostic testing of symptomatic patients to mass screening. Viewed from the perspective of United Kingdom testing, it is therefore arguable that the largest asymptomatic testing programme, by testing rate, ever attempted in the country is currently being deployed. Other countries are deploying similar programmes. Even with the advent of further waves of infection, especially associated with northern hemisphere winter, asymptomatic testing is still occurring, and it can be anticipated that as waves recede and seasons change it will again increase. These shifts have led to policies that risk conflating diagnosis with screening. Currently, there are local, regional and national variations in criteria for screening, in modes of delivery, in whether laboratories undertake confirmatory testing following positive screen results, in the extents of contact tracing undertaken, and in quality assurance of programmes, signifying discrepancies in the understanding and policy objectives of the screening being undertaken. Unavoidably, variations within the populations being tested, in terms of both disease prevalence (and hence the proportion of positive tests that are false positives) and the level of risk to the health of different individuals, also lead to implications for testing strategies, acceptability, and balancing the interests of individuals and society. Screening for SARS-CoV-2 is being undertaken with heterogeneous inclusion criteria and with heterogeneous aims [1]. The UK government's early policy stated a primary aim of making diagnoses in symptomatic individuals, with the main aim of the testing strategy being to send back to work high risk critical workers in whom a diagnosis of COVID-19 was not made [2]. However, this was rapidly extended to testing people without symptoms in care homes or returning from hospital to care homes [3,4]. More recently, there are reports of government plans to increase UK SARS-CoV-2 testing capacity to 10 million tests per day – sufficient to test the entire population each week – with these plans mentioning both symptomatic individuals and their contacts [5]. Individual organisations have used increased testing capacity to test people without symptoms with the aim of reducing transmission of SARS-CoV-2 [6,7]. Asymptomatic screening has been adopted in hospital settings to guide both infection control practices around those with higher risk of being infectious and timings of treatment for other conditions (such as elective surgery and cancer chemotherapy) where there may be a higher risk of adverse outcomes if infected [8], [9], [10], [11]. It has been adopted in community settings to allow rapid isolation and cohorting of infectious individuals in facilities and hence to reduce morbidity and mortality from institutional outbreaks [12], [13], [14]. It has even been used to monitor the progress and guide timing of containment measures for an entire town [15]. Wider screening of healthcare workers [16] and university students and staff [17] has been advocated. Testing strategies are being managed in a piecemeal fashion, but from a historical perspective this mirrors the introduction of many mass screening programmes. Heterogeneity within and between screening programmes for a single disease is not new [18,19], and as in historical cases when other screening was introduced in uncoordinated fashion, with the practice of screening ahead of evidence for its benefit, we now need to develop a systematic approach and ask to what ends we are screening, whether screening achieves these ends, and how we can approach screening methodically, in order that we can efficiently and economically achieve the best outcomes feasible as circumstances in the pandemic change. These are the purposes for which many countries have screening oversight organisations.

Breast cancer (BC) is the most frequent cancer among women worldwide and is the leading cause of cancer-related deaths in women. Cancer cells with stem cell-like features and tumor-initiating potential contribute to drug resistance, tumor recurrence, and metastasis. To achieve better clinical outcomes, it is crucial to eradicate both bulk BC cells and breast cancer stem cells (BCSCs). Salinomycin, a monocarboxylic polyether antibiotic isolated from Streptomyces albus, can precisely kill cancer stem cells (CSCs), particularly BCSCs, by various mechanisms, including apoptosis, autophagy, and necrosis. There is increasing evidence that salinomycin can inhibit cell proliferation, invasion, and migration in BC and reverse the immune-inhibitory microenvironment to prevent tumor growth and metastasis. Therefore, salinomycin is a promising therapeutic drug for BC. In this review, we summarize established mechanisms by which salinomycin protects against BC and discuss its future clinical applications.

PurposeThymic epithelial tumors (TETs) are relatively rare neoplasms, including thymomas (types A, AB, B1, B2, and B3) and thymic carcinomas (TCs). The current knowledge about the biological properties of TETs is limited due to their low incidence. This study aimed to detect genetic alterations in TETs using next-generation sequencing(NGS) and explore their clinical significance in survival.MethodsTumor tissues and clinical data were collected from 34 patients with resected TETs in the Tianjin Medical University General Hospital between January 2011 and January 2019, and 56 cancer-associated genes were analyzed. The data of 123 TETs were retrieved from TCGA, and the information on their clinical and somatic mutations was explored.ResultsThe cohort comprised 34 TETs including 17 thymomas and 17 TCs. The NGS results indicated that 73.08% of TCs+type B3 TETs and 37.50% of non-TCs+type B3 TETs each exhibited gene mutations. For patients with type B3/C, TP53 was the most frequent mutation (19.23%), followed by CDKN2A (11.54%). Similarly, in 123 TETs from the TCGA cohort, TP53 mutations were more frequent in patients with type B3/C than in patients with non-type B3/C (11.53% vs 3.09%). Further, patients with TET with TP53 mutations in the present cohort and the TCGA cohort had a worse prognosis compared with those without TP53 mutations.ConclusionsGene mutation profiles between TCs+type B3 TETs and non-TCs+type B3 TETs were significantly different. The presence of TP53 mutations was more frequent in TCs+type B3 TETs than in non-TCs+type B3 TETs, which was associated with a worse prognosis.

Muscle-invasive bladder cancer (MIBC) is an aggressive and heterogeneous malignancy. Tumor-associated macrophages (TAMs) are key infiltrating cell populations in the inflammatory microenvironment of malignant tumors including MIBC. It intrigues us to explore the clinical significance and immunoregulatory role of TAMs infiltration and polarization in MIBC. A total of 141 patients with MIBC from Zhongshan Hospital and 391 patients with MIBC from The Cancer Genome Atlas (TCGA) database were included in this study. Moreover, 195 patients who received anti-PD-L1 therapy from the IMvigor210 trial were enrolled. Patients were categorized into three subtypes considering the infiltration level and polarization status of TAMs, denoted as TAMlow (Subtype I), TAMhigh&M2/M1low (Subtype II), and TAMhigh&M2/M1high (Subtype III). Subtype III suffered inferior prognosis, and Subtype II could benefit more from adjuvant chemotherapy (ACT). Subtype III was featured with increased pro-tumor cells and immunosuppressive cytokines, while Subtype II possessed more immunogenic cells infiltration with activated and tumoricidal properties. Subtype II and Subtype III presented basal/squamous-like characterization and showed additional prognostic merit beyond molecular classification. Subtype I exhibited elevated level of FGFR3 signature, while Subtype II had EGFR signaling activation and immunotherapeutic indication. Additionally, Subtype II patients were indeed highly sensitive to PD-L1 blockade therapy in IMvigor210 trial. The infiltration and polarization status of TAMs shaped distinct immune microenvironment with predictive significance for survival outcome, ACT benefit, and PD-L1 blockade therapy sensitivity in MIBC. Immune classification based on TAMs polarization and infiltration might provide tools to tailor chemotherapy and immunotherapy.

Circulating Tumor Cells (CTCs) are cancer cells that split away from the primary tumor and appear in the circulatory system as singular units or clusters, which was first reported by Dr. Thomas Ashworth in 1869. CTCs migrate and implantation occurs at a new site, in a process commonly known as tumor metastasis. In the case of breast cancer, the tumor cells often migrate into locations such as the lungs, brain, and bones, even during the early stages, and this is a notable characteristic of breast cancer. Survival rates have increased significantly over the past few decades because of progress made in radiology and tissue biopsy, making early detection and diagnosis of breast cancer possible. However, liquid biopsy, particularly that involving the collection of CTCs, is a non-invasive method to detect tumor cells in the circulatory system, which can be easily isolated from human plasma, serum, and other body fluids. Compared to traditional tissue biopsies, fluid sample collection has the advantages of being readily available and more acceptable to the patient. It can also detect tumor cells in blood earlier and in smaller numbers, possibly allowing for diagnosis prior to any tumor detection using imaging methods. Because of the scarcity of CTCs circulating in blood vessels (only a few CTCs among billions of erythrocytes and leukocytes), thorough but accurate detection methods are particularly important for further clinical applications.

DDX60, an interferon (IFN)-inducible gene, plays a promotional role in many tumors. However, its function in glioma remains unknown. In this study, bioinformatic analysis (TCGA, CGGA, Rembrandt) illustrated the upregulation and prognostic value of DDX60 in gliomas. Immunohistochemical staining of clinical samples (n = 49) validated the DDX60 expression is higher in gliomas than in normal tissue (n = 20, P < 0.0001). It also could be included in nomogram as a parameter to predict the 3- and 5-year survival risk (C-index = 0.86). The biological process of DDX60 in glioma was mainly enriched in the inflammatory and immune response by GSEA and GO analysis. DDX60 expression had a positive association with most inflammatory-related functions, such as hematopoietic cell kinase (HCK) (R = 0.31), interferon (R = 0.72), STAT1 (R = 54), and a negative correlation with IgG (R = −0.24). Furthermore, DDX60 expression tends to be positively related to multiple infiltrating immune cells, while negatively related to CD56 dim nature killer cell in glioma. Some important immune checkpoints, like CTLA-4, PD-L1, EGF, CD96, and CD226, were all positively related with DDX60 (all Pearson correlation R > 0.26). The expression and correlation between DDX60, EGF, and PD-L1 were confirmed by western blot in clinical samples (n = 14, P < 0.0001) and GBM cells. These results indicated that DDX60 might have important clinical significance in glioma and could serve as a potential immune therapeutic target.

Background PLEKHA5 has previously been identified as a novel molecule implicated in melanoma brain metastasis, a disease that continues to portend a poor prognosis. The aim of this study was to further investigate the functional role of PLEKHA5 in disseminated melanoma. Methods The impact of PLEKHA5 on proliferation and tumor growth was examined in vitro and in melanoma xenograft models, including brain-tropic melanomas (melanomas tending to disseminate to the brain). In vitro loss- and gain-of-function studies were used to explore the underlying mechanisms of PLEKHA5-mediated tumor growth and the crosstalk between PLEKHA5 and PI3K/AKT/mTOR or MAPK/ERK signaling. The clinical relevance of PLEKHA5 dysregulation was further investigated in a cohort of matched cranial and extracranial melanoma metastases. Results PLEKHA5 stable knockdown negatively regulated cell proliferation by inhibiting the G1 -to-S cell cycle transition, which coincided with upregulation of the cell cycle regulator PDCD4. Conversely, ectopic PLEKHA5 expression exhibited the inverse effect. PLEKHA5 knockdown significantly inhibited tumor growth, whereas its overexpression upregulated the growth of tumors, which was induced by cranial and subcutaneous inoculation of cells in nude mice. PLEKHA5 modulation affected PDCD4 protein stability and was coupled with changes in PI3K/AKT/mTOR pathway signaling. High PDCD4 expression in cerebral specimens was associated with better overall survival. Conclusions This study further supports the role of PLEKHA5 as a regulator of melanoma growth at distant sites, including the brain. Furthermore, the results highlight the significance of PDCD4 dysregulation in disseminated melanoma and implicate PDCD4 as a possible causal link between PLEKHA5 and cell proliferation and growth.

Aim This study aimed to assess the clinical effectiveness of video-assisted thoracoscopic surgery (VATS) in early-stage lung cancer by indocyanine green (ICG) for tumor mapping. Material and methods Thirty patients with early-stage lung cancer with peripheral nodules smaller than 2 cm scheduled for computed tomography (CT)-guided microcoil placement followed by ICG tumor mapping by VATS wedge resection were enrolled. After microcoil deployment, 100 to 150 ml of diluted ICG was injected percutaneously near the nodule. The nodule initially was localized solely by using a near-infrared ray (NIR) thoracoscope to visualize ICG fluorescence. Thoracoscopic instruments were used to determine the staple line. Wedge resection was performed after confirmation of the location of the microcoil using fluoroscopy and pathology results. Results Thirty patients underwent VATS resection. The median tumor size was 1.3 cm by CT. The median depth from the pleural surface was 1.7 cm (range: 0.5-3.8 cm). The median CT-guided intervention time was 25 min, and VATS procedural time was 50 min. ICG fluorescence was clearly identified in 30 of 30 patients (100%). The surgical margins were all negative on final pathology in all included cases. The final diagnoses were 30 primary lung cancers; none needed additional resection. Conclusions CT-guided percutaneous ICG injection and intraoperative NIR localization of small nodules are safe and feasible. These offer surgeons the ease of localization through direct indocyanine green fluorescence imaging without the use of fluoroscopy and may be a complementary technique to preoperative microcoil placement for nonvisible, nonpalpable intrapulmonary nodules.

Objectives The aim of this study was to investigate associations between baseline characteristics and CD4 cell count response on first-line antiretroviral therapy and risk of virological failure (VF) with or without drug resistance. Methods We conducted an analysis of UK Collaborative HIV Cohort data linked to the UK HIV Drug Resistance Database. Inclusion criteria were viral sequence showing no resistance prior to initiation of first-line efavirenz + tenofovir disoproxil fumarate + emtricitabine and virological suppression within 6 months. Outcomes of VF (≥200 copies/mL) with or without drug resistance were assessed using a competing risks approach fitted jointly with a model for CD4 cell count recovery. Hazard ratios for each VF outcome were estimated for baseline CD4 cell count and viral load and characteristics of CD4 cell count response using latent variables on a standard normal scale. Results A total of 3640 people were included with 338 VF events; corresponding viral sequences were available in 134 with ≥1 resistance mutation in 36. VF with resistance was associated with lower baseline CD4 (0.30, 0.09–0.62), lower CD4 recovery (0.04, 0.00–0.17) and higher CD4 variability (4.40, 1.22–12.68). A different pattern of associations was observed for VF without resistance, but the strength of these results was less consistent across sensitivity analyses. Cumulative incidence of VF with resistance was estimated to be >2% at 3 years for baseline CD4 ≥350 cells/μL. Conclusion Lower baseline CD4 cell count and suboptimal CD4 recovery are associated with VF with drug resistance. People with low CD4 cell count before ART or with suboptimal CD4 recovery on treatment should be a priority for regimens with high genetic barrier to resistance.

This study developed an open-tubular capillary electrochromatography protocol for the analysis of antipyretic analgesic drugs, which used a multifunctional homopolymer as coating. A controlled/living radical polymerization strategy was adopted to obtain poly(N-acryloxysuccinimide) with a tunable chain-length. The homopolymer coating enhanced the separation performance by contributing to the hydrophobic and hydrogen-bonding interactions between the analytes and the homopolymer. The effect of polymer chain-length and buffer pH and concentration on the separation efficiency was evaluated. In this approach, baseline separation of the three test drugs was achieved within 15 min. The repeatability of the prepared homopolymer coating was investigated, with the relative standard deviations < 2.88% observed in intra- and interday runs. Good linearity in the 5-800 µM range (R2 ≥ 0.998) demonstrates that accurate quantitative analysis of real samples was achieved. Moreover, the proposed assay was used to quantify the three drugs (aminopyrine, 4-aminoantipyrine, and phenacetin) in urine samples, achieving recovery rates between 92.1 and 108.7%. This promising methodology may be used for the analysis of drugs in real bio-samples and for the development of unique homopolymer coatings for open-tubular capillary electrochromatography systems.

To compare the efficacy of omentoplasty with non-omentoplasty in the prevention of postoperative anastomotic leakage, and to investigate the safety of omentoplasty. Literature searches were performed of the Medline, EMBASE, and Cochrane Library databases. Studies that compared the efficacy of omentoplasty and non-omentoplasty after esophagectomy were selected. A meta-analysis was performed on anastomotic leakage, anastomotic stenosis, hospital mortality, and length of hospital stay. Results were reported as odds ratio (OR), weighted mean difference (WMD), or relative risk (RR), with 95% confidence intervals. Six studies involving a total of 1608 patients met inclusion criteria. Compared with the non-omentoplasty group, the incidence of anastomotic leakage in the omentoplasty group (OR, 0.37; 95% CI, 0.23–0.60; P

Animal-Computer Interaction (ACI) is a discipline that encourages study on animal interactions, and there are few studies that discuss disabled animals. This study will focus on cats with hearing loss and explore ways to build the emotional connection between humans and cats through interactions. The prototype design consisted of three parts, all of which were used in a two-week initial feasibility test. In the test with volunteers, two deaf cats and their keepers have participated in the interactive progress, and the owners’ feedback showed some valuable results, which may refer to further studies around this topic.

Background Non-alcoholic fatty liver disease (NAFLD) has been reported to be helpful to identify high-risk individuals of developing prostate cancer. In the retrospective cohort study, our aim is to investigate the relationship between NAFLD and biochemical recurrence in metastatic prostate cancer patients. Methods We retrospectively investigated 602 patients with metastatic prostate cancer receiving androgen deprivation therapy. Liver fat was estimated with liver-to-spleen ratio by CT scans. The relationship between NAFLD and biochemical recurrence was investigated with Cox models. The model for biochemical recurrence was adjusted for multiple variables. Results NAFLD was significantly associated with biochemical recurrence in patients with Gleason score ≥ 4 + 3 when adjusting for each of body mass index (hazards ratio [HR] = 1.38; 95% confidence interval [CI] = 1.08–1.77; p = 0.01), visceral adipose tissue (HR = 1.36; 95% CI = 1.07–1.74; p = 0.01), hypertension (HR = 1.41; 95% CI = 1.10–1.80; p = 0.01) and diabetes mellitus (HR = 1.42; 95% CI = 1.11–1.82; p = 0.01), using age and prostate-specific antigen level as potential confounder. The 2-year biochemical recurrence rate in Gleason score ≥ 4 + 3 patients with and without NAFLD was 84.0% (100/119) and 72.2% (130/180), respectively (p = 0.018). The median biochemical recurrence free survival of Gleason score ≥ 4 + 3 patients with and without NAFLD were 17 and 21 months, respectively (p = 0.005). Conclusions NAFLD is an independent risk factor for biochemical recurrence in patients with high-grade metastatic prostate cancer. If validated in prospective studies, future research should test whether treatment of NAFLD can lead to better prognosis.

Nucleic acid amplification for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in respiratory samples is the standard method for diagnosis. The majority of this testing is centralized and therefore has turnaround times of several days. Point-of-care (POC) testing with rapid turnaround times would allow more effective triage in settings where patient management and infection control decisions need to be made rapidly. The inclusivity and specificity of the Simple AMplification-Based Assay (SAMBA) II SARS-CoV-2 test were determined by both in silico analyses of the primers and probes and wet testing., Nucleic acid amplification for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in respiratory samples is the standard method for diagnosis. The majority of this testing is centralized and therefore has turnaround times of several days. Point-of-care (POC) testing with rapid turnaround times would allow more effective triage in settings where patient management and infection control decisions need to be made rapidly. The inclusivity and specificity of the Simple AMplification-Based Assay (SAMBA) II SARS-CoV-2 test were determined by both in silico analyses of the primers and probes and wet testing. The SAMBA II SARS-CoV-2 test was evaluated for performance characteristics. Clinical performance was evaluated in residual combined throat/nose swabs and compared to that of the Public Health England real-time PCR assay targeting the RdRp gene. The SAMBA II SARS-CoV-2 test has an analytical sensitivity of 250 copies/ml for detecting two regions of the genome (open reading frame 1ab [ORF1ab] and nucleocapsid protein [N]). The clinical performance was evaluated in 172 residual combined nose/throat swabs provided by the Clinical Microbiology and Public Health Laboratory, Addenbrooke’s Hospital, Cambridge (CMPHL), which showed an estimated positive percent agreement of 98.9% (95% confidence interval [CI], 93.83 to 99.97) and negative percent agreement of 96.4% (95% CI, 89.92 to 99.26) compared to testing by the CMPHL. The data show that the SAMBA II SARS-CoV-2 test performs equivalently to the centralized testing methods, but with a shorter turnaround time of 86 to 101 min. Point-of-care tests such as SAMBA should enable rapid patient management and effective implementation of infection control measures.

Highlights • A high prevalence of probable PTSD (20.87%) was found about six months after the first local outbreak of COVID-19 among healthcare workers at the Central Hospital of Wuhan. • Healthcare workers with negative COVID-19 tests, those with high Social Support Self-Rating Scale (SSRS) scores, and HCWs whose family members tested negative were less likely to have probable PTSD. • High levels of psychiatric and somatic illness and insomnia were associated with probable PTSD., This study examined the prevalence of posttraumatic stress disorder (PTSD) symptoms and assessed mental illness via an online survey among healthcare workers (HCWs) at the Central Hospital of Wuhan after the peak of the COVID-19 outbreak. PTSD symptoms were measured using the PTSD Checklist Civilian Version (PCL-C), with a cutoff score of 50. Among the 642 HCWs, the prevalence of probable PTSD was 20.87%. Additionally, 88.88%, 82.09%, 100%, and 95.52% of HCWs with probable PTSD reported varying degrees of anxiety, depression, somatic symptoms, and insomnia, respectively. HCWs with probable PTSD scored higher on the Hospital Anxiety and Depression Scale (HADS), Patient Health questionnaire-15 (PHQ-15), and Insomnia Severity Index (ISI) than non-PTSD HCWs (all p

BackgroundThe disease addiction to chemical substances or drugs such as alcohol (ethanol, C2H5OH) is still not recognised by many medical practitioners as a diagnostic nosology in Ghana. It is usually considered a moral challenge, rather than a medical condition. This perception is propagated even in the Ministry of Health’s Occupational Health and Safety (OHS) policy document which punishes symptoms of substance use disorders. This study seeks to explore the perception and attitude of employers towards employees with alcohol use disorders (AUDs).MethodA qualitative approach of in-depth interviews and observations which described the perceived attitudes of employers (using HR managers as proxy) towards employees with AUD of ten private and public hospitals in the Greater-Accra Metropolis was adopted. The face-to face interviews conducted using a an interview guide were audio-recorded and later transcribed. On average, the interview lasted between 45 and 60 minutes.ResultsFindings from the 10 interviews revealed that psychological factors accounted for AUD regardless of the nature of their work. In spite of prevalence of AUD in some hospitals, it was interesting to know that majority of these facilities had no employee assistance programmes (EAPs) and the few that had some policies on AUD were not implementing them.ConclusionThis paper focused on some aspect of HR managers’ perceived attitude toward an aspect of their mental well-being which is AUD. However, there are other pertinent issues that are crucial to employee total well-being that can be addressed in future research.

Surgery is the first line treatment for congenital concealed penis but penile retraction is inevitable in some cases. We investigate the anatomical and histological characteristics of penile fasciae and describe a new technique for the correction of concealed penis.The anatomical structures of penile fasciae were observed in 10 adult cadaveric penises. Penile tissue samples were stained with hematoxylin-eosin, Masson's trichrome and Weigert's resorcin-fuchsin, respectively. From January 2017 to May 2019, 78 patients with congenital concealed penis were treated with the new surgical technique. Median patient age was 14 years (range 8 to 18).Dartos fascia had sublayers. The superficial layer was a well vascularized tissue composed of nonpolar collagen fibers intermixed with nerves and vessels. The deep layer was composed of a transverse arrangement of collagen fibers and elastic fibers, and there were fewer venules and nerve fibers. Based on this finding we performed anatomical resection of the deep layer of dartos fascia to correct concealed penis. During the operation dartos fascia was separated into 2 layers and a complete circular resection of the deep layer was made at the base of the penis. Mean followup was 14 months. All patients and their parents were satisfied with the outcomes. None of the patients underwent postoperative penile retraction.The anatomical resection of the deep layer of dartos fascia for correcting concealed penis is technically easy, safe and effective. It provides a good cosmetic appearance and functional outcomes.

Objective: Analyze and summarize the risk assessment of perioperative infection of 263 diabetic patients undergoing small bowel lateral-lateral anastomos(intestinal bypass )in the small intestine's side ,discuss the managerial methods of antimicrobial agents during the perioperative period. Methods: Retrospectively analyze the clinical data of patients who underwent laparoscopic small bowel lateral anastomosis (intestinal bypass) from January 2018 to December 2018. The patients with HBA1c ?9.0% were selected as the observation group and patients with HBA1c < 9.0% as the control group. Results: The decling range of the average value about blood glucose in the observation group was greater than that in the control group. The difference was statistically significant (P

Summary There is urgent need for rapid SARS-CoV-2 testing in hospital to limit nosocomial spread. We report an evaluation of point of care (POC) nucleic acid amplification testing (NAAT) in 149 participants with parallel combined nasal/throat swabbing for POC versus standard lab RT-PCR testing. Median time to result is 2.6 (IQR 2.3 to 4.8) versus 26.4 hours (IQR 21.4 to 31.4, p, Graphical Abstract, Highlights • Point of care (POC) test highly sensitive and specific against RT-PCR • Time to result 2.6hrs for POC v 26.4hrs for standard lab RT-PCR. • SARS-CoV-2 POC test reduces median time-to-bed placement by approximately six hours • SARS-CoV-2 POC improves indices of hospital functioning and patient care, Collier et al report testing and implementation of an accurate SARS-CoV-2 point of care nucleic acid amplification test that leads to faster time to definitive bed placement from the ER, release of isolation rooms, avoidance of bed closures, and expedited discharges to care homes and access to hospital investigations/procedures.

Introduction. Pneumonia is highly prevalent in intensive care units (ICUs), with high associated mortality. Empirical treatment prioritizes breadth of coverage while awaiting laboratory diagnosis, often at the expense of antimicrobial stewardship. Microarrays use multiple parallel polymerase chain reactions to enable a rapid syndromic approach to laboratory diagnosis. Aim. To evaluate the clinical and laboratory implications of introducing a bespoke 22-pathogen TaqMan Array Card (TAC) for rapid pathogen detection in deep respiratory samples from adult ICUs. Methodology. TAC results from all ICU patients prospectively tested over a 9-month period at Cambridge’s Clinical Microbiology and Public Health Laboratory were compared to those of corresponding conventional microbiological assays (culture-, PCR- or serology-based) in terms of result agreement and time-to-result availability. Clinical impact was assessed by retrospective review of medical records. Results. Seventy-one patients were included [45 (63 %) male, median age 59). Overall result agreement was 94 %, with TAC detecting more pathogens than conventional methods. TAC detected Streptococcus pneumoniae more readily than culture (7 vs 0 cases; P=0.02). TAC did not detect Aspergillus spp. in eight culture- or galactomannan-positive cases. The median turnaround time (1 day) was significantly shorter than that of bacterial/fungal culture, Pneumocystis jirovecii PCR and galactomannan testing (each 3 days; PP Mycobacterium tuberculosis culture (46 days; P Conclusion. TAC provided greater overall yield of pathogen detection and faster turnaround times, permitting earlier discontinuation of unnecessary antimicrobials.

BackgroundRapid COVID-19 diagnosis in hospital is essential for patient management and identification of infectious patients to limit the potential for nosocomial transmission. The diagnosis of infection is complicated by 30-50% of COVID-19 hospital admissions with nose/throat swabs testing negative for SARS-CoV-2 nucleic acid, frequently after the first week of illness when SARS-CoV-2 antibody responses become detectable. We assessed the diagnostic accuracy of combined rapid antibody point of care (POC) and nucleic acid assays for suspected COVID-19 disease in the emergency department.MethodsWe developed (i) an in vitro neutralization assay using a lentivirus expressing a genome encoding luciferase and pseudotyped with spike (S) protein and (ii) an ELISA test to detect IgG antibodies to nucleocapsid (N) and S proteins from SARS-CoV-2. We tested two lateral flow rapid fingerprick tests with bands for IgG and IgM. We then prospectively recruited participants with suspected moderate to severe COVID-19 and tested for SARS-CoV-2 nucleic acid in a combined nasal/throat swab using the standard laboratory RT-PCR and a validated rapid POC nucleic acid amplification (NAAT) test. Additionally, serum collected at admission was retrospectively tested by in vitro neutralisation, ELISA and the candidate POC antibody tests. We evaluated the performance of the individual and combined rapid POC diagnostic tests against a composite reference standard of neutralisation and standard laboratory based RT-PCR.Results45 participants had specimens tested for nucleic acid in nose/throat swabs as well as stored sera for antibodies. Using the composite reference standard, prevalence of COVID-19 disease was 53.3% (24/45). Median age was 73.5 (IQR 54.0-86.5) years in those with COVID-19 disease by our reference standard and 63.0 (IQR 41.0-72.0) years in those without disease. The overall detection rate by rapid NAAT was 79.2% (95CI 57.8-92.9%), decreasing from 100% (95% CI 65.3-98.6%) in days 1-4 to 50.0% (95% CI 11.8-88.2) for days 9-28 post symptom onset. Correct identification of COVID-19 with combined rapid POC diagnostic tests was 100% (95CI 85.8-100%) with a false positive rate of 5.3-14.3%, driven by POC LFA antibody tests.ConclusionsCombined POC tests have the potential to transform our management of COVID-19, including inflammatory manifestations later in disease where nucleic acid test results are negative. A rapid combined approach will also aid recruitment into clinical trials and in prescribing therapeutics, particularly where potentially harmful immune modulators (including steroids) are used.

Nucleic acid amplification for the detection of SARS-CoV-2 RNA in respiratory samples is the standard method for diagnosis. These tests are centralised and therefore turnaround times can be 2-5 days. Point-of-care testing with rapid turnaround times would allow more effective triage in settings where patient management and infection control decisions need to be made rapidly.Inclusivity and specificity of the SAMBA II SARS-CoV-2 assay was determined by in silico analyses of the primers and probes. Analytical and clinical sensitivity and specificity of the SAMBA II SARS-CoV-2 Test was evaluated for analytical sensitivity and specificity. Clinical performance was evaluated in residual clinical samples compared to the Public Health England reference tests.The limit of detection of the SAMBA II SARS-CoV-2 Test is 250 cp/mL and is specific for detection of 2 regions of the SARS-CoV-2 genome. The clinical sensitivity was evaluated in 172 clinical samples provided by the Clinical Microbiology and Public Health Laboratory, Addenbrooke’s Hospital, Cambridge (CMPHL), which showed a sensitivity of 98.9% (95% CI 94.03-99.97%), specificity of 100% (95% CI 95.55-100%), PPV of 100% and NPV of 98.78% (92.02-99.82%) compared to testing by CMPHLSAMBA detected 3 positive samples that were initially negative by PHE Test. The data shows that the SAMBA II SARS-CoV-2 Test performs equivalently to the centralised testing methods with a much quicker turnaround time. Point of care testing, such as SAMBA, should enable rapid patient management and effective implementation of infection control measures.

Summary Increased neoantigens in hypermutated cancers with DNA mismatch repair deficiency (dMMR) are proposed as the major contributor to the high objective response rate in anti-PD-1 therapy. However, the mechanism of drug resistance is not fully understood. Using tumor models defective in the MMR gene Mlh1 (dMLH1), we show that dMLH1 tumor cells accumulate cytosolic DNA and produce IFN-β in a cGAS-STING-dependent manner, which renders dMLH1 tumors slowly progressive and highly sensitive to checkpoint blockade. In neoantigen-fixed models, dMLH1 tumors potently induce T cell priming and lose resistance to checkpoint therapy independent of tumor mutational burden. Accordingly, loss of STING or cGAS in tumor cells decreases tumor infiltration of T cells and endows resistance to checkpoint blockade. Clinically, downregulation of cGAS/STING in human dMMR cancers correlates with poor prognosis. We conclude that DNA sensing within tumor cells is essential for dMMR-triggered anti-tumor immunity. This study provides new mechanisms and biomarkers for anti-dMMR-cancer immunotherapy.

The cell membrane is composed of a phospholipid bilayer with embedded proteins and maintains cell homeostasis through dynamic changes. An abnormal cell membrane shape could be a sign of unhealthy cells. Probes for subcellular fluorescence imaging that can identify the abnormal plasma membrane and record the dynamic changes are needed. Based on a solvatochromic dye with a near-infrared emission strategy, the amphipathic molecule (E)-2,2'-((4-(2-(4-(dicyanomethylene)-4H-chromen-2-yl)vinyl)phenyl)azanediyl)bis(ethane-1-sulfonic acid) (MRL) contained a hydrophilic sulfo group and a hydrophobic chromone group, which was designed and synthesized for staining the cell membrane and monitoring the morphology of the membranes under different conditions. MRL exhibited an excellent photostability and low cytotoxicity; when cells were incubated with MRL, cell membranes were specifically labeled. MRL is capable of long-term monitoring of the morphological changes of cell membrane.