Search

Abstract Background This study aimed to investigate whether persistent human papillomavirus integration at the same loci (PHISL) before and after treatment can predict recurrent/residual disease in women with CIN2-3. Methods A total of 151 CIN2-3 women treated with conization between August 2020 and September 2021 were included. To investigate the precision of HPV integration, we further analyzed HPV integration-positive patients. Sensitivity, specificity, positive and negative predictive values (PPV and NPV, respectively), and the Youden index for predicting recurrence/residual disease were calculated. Results Among the 151 enrolled CIN2-3 women, 56 were HPV integration-positive and 95 had HPV integration-negative results. Six (10.7%) experienced recurrence among 56 HPV integration-positive patients, which was more than those in HPV integration-negative patients (one patient, 1.1%). In the 56 HPV integration-positive patients, 12 had positive HPV results after treatment, seven had PHISL, and two had positive cone margin. Among the seven patients who tested with PHISL, six (85.7%) had residual/recurrent disease. PHISL was a prominent predictor of persistent/recurrent disease. The HPV test, the HPV integration test, and PHISL all had a sensitivity of 100% and a NPV of 100% for residual/recurrent disease. PHISL showed better specificity (98.0% vs. 82.0%, p = 0.005) and PPV (85.7% vs. 40.0%, p = 0.001) than the HPV test for predicting recurrence. Conclusions The HPV-integration-positive CIN2-3 women had much higher relapse rates than HPV-integration-negative CIN2-3 women. The findings indicate that PHISL derived from preoperative and postoperative HPV integration tests may be a precise biomarker for the identification of residual/recurrent CIN 2/3.

Abstract Background Ferroptosis is an iron-dependent type of regulated cell death, and has been implicated in lung adenocarcinoma (LUAD). Evidence has proved the key role of glutamate-cysteine ligase catalytic subunit (GCLC) in ferroptosis, but its role in LUAD remains unclear. Herein, we explored the implications of GCLC and relevant genes in LUAD prognosis and immunity as well as underlying molecular mechanisms. Methods This work gathered mRNA, miRNA, DNA methylation, somatic mutation and copy-number variation data from TCGA-LUAD. WGCNA was utilized for selecting GCLC-relevant genes, and a GCLC-relevant prognostic signature was built by uni- and multivariate-cox regression analyses. Immune compositions were estimated via CIBERSORT, and two immunotherapy cohorts of solid tumors were analyzed. Multi-omics regulatory mechanisms were finally assessed. Results Our results showed that GCLC was overexpressed in LUAD, and potentially resulted in undesirable survival. A prognostic model was generated, which owned accurate and independent performance in prognostication. GCLC, and relevant genes were notably connected with immune compositions and immune checkpoints. High GCLC expression was linked with better responses to anti-PD-L1 and anti-CTLA-4 treatment. Their possible DNA methylation sites were inferred, e.g., hypomethylation in cg19740353 might contribute to GCLC up-regulation. Frequent genetic mutations also affected their expression. Upstream transcription factors (E2F1/3/4, etc.), post-transcriptional regulation of miRNAs (hsa-mir-30c-1, etc.), lncRNAs (C8orf34-AS1, etc.), and IGF2BP1-mediated m6A modification were identified. It was also found NOP58-mediated SUMOylation post-translational modification. Conclusions Together, we show that GCLC and relevant genes exert crucial roles in LUAD prognosis and immunity, and their expression can be controlled by complex multi-omics mechanisms.

Urbanization has facilitated economic development while simultaneously resulting in various ecological issues. Constructing a multi-scale nested and composite functional urban-rural ecological network is crucial for improving ecological security. This study utilizes Dali City as a case study and employs methods including MSPA, circuit theory, and landscape connectivity index to develop the urban-rural habitat network, water green network, and recreation network, focusing on the '' red-green-blue '' spatial framework. An analysis of the spatial characteristics of source areas, corridors, ecological strategic points, and other spatial elements is conducted to establish a multi-level, multi-objective, and multifunctional composite urban-rural ecological network. The results show that: (1) 13 ecological source areas were identified in both the municipal and main urban areas, along with 22 ecological corridors in the municipal and 20 main urban areas. The distribution of ecological corridors was uniform across the study area. (2) The optimal width for the municipal biological corridor is 150 m, the main urban area should have a width of 90 m. The optimal width for rainwater corridors in municipal and main urban areas is 60 m. (3) The multi-scale nested ecological network identified 4 common ecological sources, 11 ecological corridors, 3 rainwater corridors, 6 wetland nodes, and 7 amusement nodes. Overall, the number of ecological nodes is limited, indicating a need for enhanced node construction. The research findings offer insights for developing ecological networks that integrate urban and rural functions, serving as a reference for ecological protection and restoration in pertinent regions.

IntroductionCognitive impairment is a frequent clinical symptom of non-communicating hydrocephalus (NCH) involving multiple domains, including executive function, working memory, visual-spatial function, language, and attention. Functional magnetic resonance imaging (fMRI) can be used to obtain information on functional activity in local brain areas and functional connectivity (FC) across multiple brain regions. However, studies on the associated cognitive impairment are limited; further, the pathophysiological mechanisms of NCH with cognitive impairment remain unclear. Here, we aimed to explore alterations in regional neural activity and FC, as well as the mechanisms of cognitive impairment, in patients with NCH.MethodsOverall, 16 patients with NCH and 25 demographically matched healthy controls (HCs) were assessed using the Mini-Mental State Examination (MMSE) and fMRI. Changes in regional homogeneity (ReHo), degree centrality (DC), and region of interest-based FC were analyzed in both groups. The relationship between fMRI metrics (ReHo, DC, and FC) and MMSE scores in patients with NCH was also investigated.Results and discussionCompared with the HC group, the NCH group exhibited significantly lower ReHo values in the left precentral and postcentral gyri, and significantly higher ReHo values in the left medial prefrontal cortex (MPFC). The NCH group also showed significantly higher DC values in the bilateral MPFC compared with the HC group. Regarding seed-based FC, the MPFC showed reduced FC values in the right superior parietal and postcentral gyrus in the NCH group compared with those in the HC group. Moreover, within the NCH group, MMSE scores were significantly negatively correlated with the ReHo value in the left MPFC and the DC value in the bilateral MPFC, whereas MMSE scores were significantly positively correlated with FC values. To conclude, regional neural activity and FC are altered in patients with NCH and are correlated with cognitive impairment. These results advance our understanding of the pathophysiological mechanisms underlying the association between NCH and cognitive impairment.

Abstract Background Cervical cancer (CC) causes more than 311,000 deaths annually worldwide. The integration of human papillomavirus (HPV) is a crucial genetic event that contributes to cervical carcinogenesis. Despite HPV DNA integration is known to disrupt the genomic architecture of both the host and viral genomes in CC, the complexity of this process remains largely unexplored. Results In this study, we conducted whole-genome sequencing (WGS) at 55-65X coverage utilizing the PacBio long-read sequencing platform in SiHa and HeLa cells, followed by comprehensive analyses of the sequence data to elucidate the complexity of HPV integration. Firstly, our results demonstrated that PacBio long-read sequencing effectively identifies HPV integration breakpoints with comparable accuracy to targeted-capture Next-generation sequencing (NGS) methods. Secondly, we constructed detailed models of complex integrated genome structures that included both the HPV genome and nearby regions of the human genome by utilizing PacBio long-read WGS. Thirdly, our sequencing results revealed the occurrence of a wide variety of genome-wide structural variations (SVs) in SiHa and HeLa cells. Additionally, our analysis further revealed a potential correlation between changes in gene expression levels and SVs on chromosome 13 in the genome of SiHa cells. Conclusions Using PacBio long-read sequencing, we have successfully constructed complex models illustrating HPV integrated genome structures in SiHa and HeLa cells. This accomplishment serves as a compelling demonstration of the valuable capabilities of long-read sequencing in detecting and characterizing HPV genomic integration structures within human cells. Furthermore, these findings offer critical insights into the complex process of HPV16 and HPV18 integration and their potential contribution to the development of cervical cancer.

BackgroundNormal pressure hydrocephalus (NPH) has drawn an increasing amount of attention over the last 20 years. At present, there is a shortage of intuitive analysis on the trends in development, key contributors, and research hotspots topics in the NPH field. This study aims to analyze the evolution of NPH research, evaluate publications both qualitatively and quantitatively, and summarize the current research hotspots.MethodsA bibliometric analysis was conducted on data retrieved from the Web of Science Core Collection (WoSCC) database between 2003 and 2023. Quantitative assessments were conducted using bibliometric analysis tools such as VOSviewer and CiteSpace software.ResultsA total of 2,248 articles published between 2003 and 2023 were retrieved. During this period, the number of publications steadily increased. The United States was the largest contributor. The University of Gothenburg led among institutions conducting relevant research. Eide P. K. was the most prolific author. The Journal of Neurosurgery is the leading journal on NPH. According to the analysis of the co-occurrence of keywords and co-cited references, the primary research directions identified were pathophysiology, precise diagnosis, and individualized treatment. Recent research hotspots have mainly focused on epidemiology, the glymphatic system, and CSF biomarkers.ConclusionThe comprehensive bibliometric analysis of NPH offers insights into the main research directions, highlights key countries, contributors, and journals, and identifies significant research hotspots. This information serves as a valuable reference for scholars to further study NPH.

Axions and axion-like particles (ALPs) appear in many extensions of the Standard Model and are being investigated as promising dark matter (DM) candidates. One viable methodology for their detection involves the investigation of the line-like radio emissions from the dwarf spheroidal galaxy, potentially originating from the radiative decay of ALPs or the conversion of ALPs in the magnetic field. In this work, we constrain the properties of ALPs using the 2-hour radio observation of Coma Berenices through the Five-hundred-meter Aperture Spherical radio Telescope (FAST). The 95% upper limits of the ALP-photon coupling are calculated for the ALP decay and conversion scenarios, respectively. Note that the sensitive ALP masses for FAST range from ∼μeV to tens of μeV, where ALP can explain the DM abundance naturally. However, our limits are weaker than those of the CAST helioscope, which can provide an independent and complementary check on the ALP non-detection for ground experiments. Furthermore, we evaluate the expected sensitivity on the ALP of FAST with its full designed bandwidth (70 MHz - 3 GHz) for 100 hours of observation time. Our results indicate that, even with the exceptional sensitivity of the FAST, it is challenging to surpass the existing experimental constraints on ALP DM using radio observation of dSphs, unless the possible enhancements of ALP signals by compact stars in dSphs are considered.

Rapid urbanization leads to fragmentation and reduced connectivity of urban landscapes, endangering regional biodiversity conservation and sustainable development. Constructing a red, green, and blue spatial ecological network is an effective way to alleviate ecological pressure and promote economic development. Using circuit theory, hydrological analysis, and suitability analysis, this study constructs a composite ecological network under urban–rural integration. The results show the following: (1) A total of 22 ecological corridors with a length of 349.20 km, 22 ecological pinch points, and 22 ecological barrier points are identified in the municipal area, mainly distributed in Haidong Town. There are 504 stormwater corridors, which are more evenly distributed, 502 riverfront landscape corridors, and 130 slow-moving landscape corridors. (2) A total of 20 ecological corridors, with a length of 99.23 km, 19 ecological pinch points, and 25 barrier points were identified in the main urban area, and most of them are located in the ecological corridors. There are 71 stormwater corridors, mainly located in the northwestern forest area, 71 riverfront recreation corridors, and 50 slow-moving recreation corridors. (3) Two scales of superimposed ecological source area of 3.65 km2, and eleven ecological corridors, are primarily distributed between Erhai Lake and Xiaguan Town. There are two superimposed stormwater corridors and fourteen recreational corridors. The eco-nodes are mostly distributed in the east and south of Dali City; wetland nodes are mainly situated in the eighteen streams of Cangshan Mountain; and landscape nodes are more balanced in spatial distribution. The study results can provide a reference for composite ecological network construction.

Combination immunotherapy of cancer vaccines with immune checkpoint inhibitors (ICIs) represents a promising therapeutic strategy for immunosuppressed and cold tumors. However, this strategy still faces challenges, including the limited therapeutic efficacy of cancer vaccines and immune-related adverse events associated with systematic delivery of ICIs. Herein, we demonstrate the antitumor immune response induced by outer membrane vesicle from Akkermansia muciniphila (Akk-OMV), which exhibites a favorable safety profile, highlighting the potential application as a natural and biocompatible self-adjuvanting vesicle. Utilizing tumor cell-derived exosome as an antigen source and Akk-OMV as a natural adjuvant, we construct a cancer vaccine formulation of extracellular vesicles hybrid lipid nanovesicles (Lipo@HEV) for enhanced prophylactic and therapeutic vaccination by promoting dendritic cell (DC) maturation in lymph node and activating cytotoxic T cell (CTL) response. The Lipo@HEV is further loaded with plasmid to enable gene therapy-mediated PD-L1 blockade upon peritumoral injection. Meanwhile, it penetrates into lymph node to initiate DC maturation and CTL activation, synergistically inhibiting the established tumor. The fabrication of extracellular vesicles hybrid plasmid-loaded lipid nanovesicles reveals a promising gene therapy-guided and vesicle-based hybrid system for therapeutic cancer vaccination and synergistic immunotherapy strategy.

This study aimed to explore effects of indica rice addition, rice soaking time and rice soup addition on total sugar and alcohol content of semi-dry Hakka rice wine (HRW) and compare its difference in physicochemical properties and volatiles with traditional sweet rice wine (TSRW) via HPLC, GC–MS and E-tongue. The optimal fermentation conditions of semi-dry HRW were 50 % indica rice addition, 12 h rice soaking time and 85 % rice soup addition. The total sugar (16.13 mg/mL) of semi-dry HRW was significantly lower than that of TSRW (135.79 mg/mL), especially the trehalose, glucose, sucrose and maltose. Its alcohol content was significantly higher than that of TSRW. There were significant differences in volatile components between semi-dry HRW and TSRW, especially esters, alcohols and ketones, but no significant differences in organic acids and amino acids. Results obtained could provide reference data for improving the fermentation process and quality of semi-dry HRW.

Stem cell based transplants effectively regenerate tissues; however, limitations such as immune rejection and teratoma formation prevent their application. Extracellular vesicles (EVs)-mediated acellular tissue regeneration is a promising alternative to stem cell based transplants. Although neural EGFL-like 1 (Nell1) is known to contribute to the osteogenic differentiation of bone marrow stem cells (BMSCs), it remains unknown whether EVs are involved in this process. Here, we present that EVs derived from Nell1-modified BMSCs (Nell1/EVs) have a stronger ability to promote BMSC osteogenesis owing to miR-25–5p downregulation. MiR-25–5p inhibits osteogenesis by targeting Smad2 and suppressing the SMAD and extracellular signal-related kinase 1 and 2 (ERK1/2) pathway activation. In addition, we demonstrate that the 3D-Nell1/EV-hydrogel system is beneficial for bone regeneration in vivo, probably stemming from a slow, continuous release and high concentration of EVs in the bone defect area. Thus, our results have shown the potential of Nell1/EVs as a novel acellular bone regeneration strategy. Mechanistically, the identification of miR-25-5p-SMAD2 signaling axis expands the knowledge of Nell1/EVs induced osteogenesis.

Background With the advancement of modern neuroimaging and neuronavigation techniques, frameless intracranial biopsy techniques have been increasingly used. Objective To assess the application value of a neuronavigation system using 3.0T magnetic resonance imaging (MRI) in frameless stereotactic brain biopsies. Methods Twenty-seven patients with intracranial lesions were recruited in the Department of Neurosurgery, People's Hospital of Xinjiang Uygur Autonomous Region from March 2018 to June 2021. All of them underwent brain MRI at 3.0T, then registered for using the Medtronic Stealth Station S7 neuronavigation system to implement frameless stereotactic brain biopsies. Pre-biopsy data, post-biopsy complications and pathological diagnosis rate were reviewed retrospectively. Results In all, 27 biopsies were performed in these 27 cases, among which anaplastic astrocytoma and large B-cell lymphoma accounted for 33.3% (9/27) , and 18.5% (5/27) , respectively. Twenty-six cases obtained a definite diagnosis by frameless stereotactic brain biopsies, with a rate of pathological diagnosis of 96.3%. During the brain biopsy, puncture tract bleeding occurred in two cases (7.4%) , and one of them was confirmed with serious bleeding by post-biopsy brain CT. Five cases had post-biopsy complications (18.5%) , four of them had intracranial hemorrhage (14.8%) , and the other one case had pulmonary infection (3.7%) . No death occurred in the biopsy. Conclusion This kind of frameless stereotactic brain biopsy is safe and effective with a high diagnostic rate.

Summary: Background: Cervical cancer is the fourth leading cause of mortality among gynecological malignancies. However, the identification of cervical cancer stem cells remains unclear. Methods: We performed single-cell mRNA sequencing on ∼122,400 cells from 20 cervical biopsies, including 5 healthy controls, 4 high-grade intraepithelial neoplasias, 5 microinvasive carcinomas of the cervix, and 6 invasive cervical squamous carcinomas. Bioinformatic results were validated by multiplex immunohistochemistry (mIHC) in cervical cancer tissue microarrays (TMA) (n = 85). Findings: We identified cervical cancer stem cells and highlighted the functional changes in cervical stem cells during malignant transformation. The original non-malignant stem cell properties (characterized by high proliferation) gradually diminished, whereas the tumor stem cell properties (characterized by epithelial-mesenchymal transformation and invasion) were enhanced. The mIHC results of our TMA cohort confirmed the existence of stem-like cells and indicated that cluster correlated with neoplastic recurrence. Subsequently, we investigated malignant and immune cell heterogeneity in the cervical multicellular ecosystem across different disease stages. We observed global upregulation of interferon responses in the cervical microenvironment during lesion progression. Interpretation: Our results provide more insights into cervical premalignant and malignant lesion microenvironments. Funding: This research was supported by the Guangdong Provincial Natural Science Foundation of China (2023A1515010382), Grant 2021YFC2700603 from the National Key Research & Development Program of China and the Hubei Provincial Natural Science Foundation of China (2022CFB174 and 2022CFB893).

Abstract The fast developments of radio astronomy open a new window to explore the properties of Dark Matter (DM). The recent direct imaging of the supermassive black hole (SMBH) at the center of M87 radio galaxy by the Event Horizon Telescope (EHT) collaboration is expected to be very useful to search for possible new physics. In this work, we illustrate that such results can be used to detect the possible synchrotron radiation signature produced by DM annihilation from the innermost region of the SMBH. Assuming the existence of a spike DM density profile, we obtain the flux density due to DM annihilation induced electrons and positrons, and derive new limits on the DM annihilation cross section via the comparison with the EHT integral flux density at 230 GHz. Our results show that the parameter space can be probed by the EHT observations is largely complementary to other experiments. For DM with typical mass regions of being weakly interacting massive particles, the annihilation cross section several orders of magnitude below the thermal production level can be excluded by the EHT observations under the density spike assumption. Future EHT observations may further improve the sensitivity on the DM searches, and may also provide a unique opportunity to test the interplay between DM and the SMBH.

To control seasonal influenza epidemics in elders, a quadrivalent, inactivated, split-virion influenza vaccine (IIV4) comprising A and B lineages is produced for young individuals and adults aged ≥60 years. In this phase III, randomized, double-blind, active-controlled trial, we compared safety and immunogenicity of IIV4 with a licensed quadrivalent inactivated vaccine (IIV4-HL) produced by Hualan Biological Engineering during the 2019 influenza season. Participants were randomly assigned to receive IIV4 (n = 959) or IIV4-HL (n = 959). Compared to IIV4-HL, geometric mean titers (GMT) of hemagglutination inhibition (HAI) titers and seroconversion rate (SCR) of IIV4 demonstrated better antibody responses in A lineages (H1N1 and H3N2) (P .01) in both age groups. After immunization, IIV4 provided a satisfactory SCR and seroprotection rate (SPR) in elders. No discernible variation in immunogenicity was observed between the two age cohorts. In both age groups, IIV4 and IIV4-HL recipients experienced similar levels of solicited and unsolicited adverse events (AEs), and the incidence of AEs was low in both vaccine groups. Most AEs were of mild-to-moderate severity and no grade 3 AEs in IIV4 group, but AEs in adults aged 60–65 were little higher than in adults over 65 years in IIV4 and IIV4-HL groups (IIV4: 14.66% vs. 10.36%; IIV4-HL:14.67% vs. 11.43%). Totally, IIV4 was generally well tolerated and induced high antibody titers against all four influenza strains in elderly, making it a compelling alternative for the elderly aged ≥60 years. Trial registration: Clinical Trials.gov: 2015L00649-2.

In recent years, the landscape ecological security of Xishuangbanna in southwest China has become an essential factor affecting the cross-border ecological security in South Asia and Southeast Asia. Based on the change of land use in Xishuangbanna, with the help of "3S" technology, landscape ecology theory, and gray prediction model, the spatial and developmental trends of landscape ecological security in Xishuangbanna from 1996-2030 could be determined. In more than 20 years, the woodland landscape area in Xishuangbanna decreased, and the fragmentation of construction land has increased overall. In 1996, the overall landscape ecological safety was good, with 63.5% of the total area of grade I and II. In 2003, the proportion of the grade I and grade II areas decreased, with landscape ecological security problems appearing. In 2010, the overall landscape ecological security area reached 74.5%, the largest proportion in more than 20 years. The grade V area accounted for only 9% and was mainly distributed on the border of Menghai County and central Jinghong City. In 2017, The grade IV and V areas was further increased, and the ecological security problem intensified. The prediction results showed that from 2023 to 2030, the regions of grades I and II increased, but the proportion of level V regions increased. Furthermore, the grade IV transformed to grade V rapidly, reaching its highest value in more than 20 years. From 1996 to 2030, the landscape ecological security space significantly evolved, showing an evident "east-south" trend in movement and eventually shifting to the southeast.

This phase III clinical trial aimed to assess the safety and demonstrate the immunogenicity of a candidate freeze-dried purified Vero cell-based rabies vaccine (PVRV-WIBP) developed for human use. A cohort of 40 participants in stage 1 and 1956 subjects in stage 2 with an age range of 10–50 years were recruited for the phase III clinical trial. For safety analysis in stage 1, 20 participants received either 4-dose or 5-dose regimen of PVRV-WIBP. In stage 2, 1956 subjects were randomly divided into the 5-dose PVRV-WIBP, 5-dose PVRV-LNCD, and 4-dose PVRV-WIBP groups. The serum neutralizing antibody titer against rabies was determined on day 7 or 14 and day 35 or 42. Adverse reactions were recorded for more than 6 months. Most adverse reactions, which were mild and moderate in severity, occurred and resolved within 1 week after each injection in the PVRV-WIBP (4 and 5 doses) and PVRV-LNCD (5 doses) groups. All three groups achieved complete seroconversion 14 days after the initial dose and 14 days after completing the full vaccination schedule, the susceptible subjects in the PVRV-WIBP group (4-dose or 5-dose regimen) displayed higher neutralizing antibody titers against the rabies virus compared to those in the PVRV-LNCD group (5-dose regimen). PVRV-WIBP induced non-inferior immune responses versus PVRV-LNCD as assessed by seroconversion rate. PVRV-WIBP was well tolerated and non-inferior to PVRV-LNCD in healthy individuals aged 10–50 years. The results indicated that PVRV-WIBP (both 4- and 5-dose schedules) could be an alternative to rabies post-exposure prophylaxis.

Galactic center is one of the most important cosmic-ray sources. Here, the authors show GeV-TeV cosmic ray density in the central molecular zone is lower than the cosmic ray sea component, suggesting presence of high energy particle accelerator at the galactic center and existence of barrier.

Background A quadrivalent split influenza vaccine IIV4-W against both influenza A and B viruses is urgently needed. Methods To evaluate the safety and immunogenicity of IIV4-W in people aged 3–60 years, 2400 participants recruited in a double-blind phase III trial and were randomly assigned to the IIV4-W, TIV1 and TIV2 groups. The immunogenicity indicators were measured at 28 days postvaccination and for 180 days for safety follow-up. Results Adverse events (AEs) occurred in 162 (20.28%), 116 (14.55%) and 123 (15.41%) participants in the IIV4-W, TIV1 and TIV2 groups, respectively. All these AEs were mild and self-limiting, and no serious AEs related to the vaccines were observed. IIV4-W elicited a non-inferior immune response for matched strains (the lower limit of 95% CI for GMT ratio >0.67, for SCR and SPR difference >-10%) and superior immune response for the additional B strains (the lower limit of 95% CI for GMT ratio >1.5, for SCR difference >10%) versus TIVs. The lower limit of the 95% confidence interval of the GMT increase fold, the seroconversion rate and the seroprotection rate exceeded 2.5, 40% and 70% for the four strains in IIV4-W respectively. Conclusions IIV4-W was noninferior to the TIV-matched strains and was superior to the additional B strain. IIV4-W was safe in the participants and elicited high antibody titers.

Abstract Background BCOR-CCNB3 sarcoma (BCS) is a group of undifferentiated small round cell sarcomas harboring the BCOR gene rearrangement which shares morphology with the Ewing sarcoma family as well as other malignant round blue cell tumors, thus making them difficult to diagnose. The aim of this study was to explore the role of molecular techniques in the diagnosis of BCS. Methods Twenty-three cases of EWSR1 rearrangement-negative undifferentiated small round cell sarcomas (Ewing-like sarcoma) were analyzed for the presence of BCOR gene rearrangement by Fluorescence in situ hybridization (FISH) and Reverse Transcription -Polymerase Chain Reaction (RT-PCR). The clinicopathological features of the positive cases were also reviewed. Fifteen additional cases were used as negative controls. Results Eight cases were found with BCOR gene rearrangement by FISH and reappraised as BCS. The patients ranged in age from 8 to 20 years old, with a male predominance (M:F = 6:2). All tumors were located in the lower extremities. The tumor locations were more common in bone (n = 6) than deep soft tissue (n = 2). Histologically, 7 of 8 tumors were predominately composed of spindle or ovoid cells. The tumor cells were usually arranged in solid hypercellular sheets without a distinct architectural pattern. IHC showed expression of TLE1 (100%), CCNB3 (88%), BCOR (71%). RT-PCR for BCOR-CCNB3 fusion transcript was positive in 7 of 8 cases. Pre-operative chemotherapy resulted in eradication of tumors in 5 patients after a follow-up of 7 to 42 months. Conclusions Efficient diagnosis of BCOR rearranged sarcomas is achieved by the using a combination of FISH and RT-PCR assays.

Abstract Purpose Critically ill COVID-19 patients have significantly increased risk of death. Although several circulating biomarkers are thought to be related to COVID-19 severity, few studies have focused on the characteristics of critically ill patients with different outcomes. The objective of this study was to perform a longitudinal investigation of the potential mechanisms affecting the prognosis of critically ill COVID-19 patients. Methods In addition to clinical data, 113 whole blood samples and 85 serum samples were collected from 33 severe and critical COVID-19 patients without selected comorbidities. Multi-omics analysis was then performed using longitudinal samples. Results Obvious transcriptional transitions were more frequent in critical survivors than in critical non-survivors, indicating that phase transition may be related to survival. Based on analysis of differentially expressed genes during transition, the erythrocyte differentiation pathway was significantly enriched. Furthermore, clinical data indicated that red blood cell counts showed greater fluctuation in survivors than in non-survivors. Moreover, declining red blood cell counts and hemoglobin levels were validated as prognostic markers of poor outcome in an independent cohort of 114 critical COVID-19 patients. Protein–metabolite–lipid network analysis indicated that tryptophan metabolism and melatonin may contribute to molecular transitions in critical COVID-19 patients with different outcomes. Conclusions This study systematically and comprehensively depicted the longitudinal hallmarks of critical COVID-19 patients and indicated that multi-omics transition may impact the prognosis. Take home message Frequent transcriptional phase transitions may contribute to outcome in critically ill COVID-19 patients. Furthermore, fluctuation in red blood cell and hemoglobin levels may relate to poor prognosis. The biological function of melatonin was suppressed in COVID-19 non-survivors, which may provide a potential theoretical basis for clinical administration.

Abstract Background The incidence rates of cervical cancer in developing countries have been steeply increasing while the medical resources for prevention, detection, and treatment are still quite limited. Computer-based deep learning methods can achieve high-accuracy fast cancer screening. Such methods can lead to early diagnosis, effective treatment, and hopefully successful prevention of cervical cancer. In this work, we seek to construct a robust deep convolutional neural network (DCNN) model that can assist pathologists in screening cervical cancer. Methods ThinPrep cytologic test (TCT) images diagnosed by pathologists from many collaborating hospitals in different regions were collected. The images were divided into a training dataset (13,775 images), validation dataset (2301 images), and test dataset (408,030 images from 290 scanned copies) for training and effect evaluation of a faster region convolutional neural network (Faster R-CNN) system. Results The sensitivity and specificity of the proposed cervical cancer screening system was 99.4 and 34.8%, respectively, with an area under the curve (AUC) of 0.67. The model could also distinguish between negative and positive cells. The sensitivity values of the atypical squamous cells of undetermined significance (ASCUS), the low-grade squamous intraepithelial lesion (LSIL), and the high-grade squamous intraepithelial lesions (HSIL) were 89.3, 71.5, and 73.9%, respectively. This system could quickly classify the images and generate a test report in about 3 minutes. Hence, the system can reduce the burden on the pathologists and saves them valuable time to analyze more complex cases. Conclusions In our study, a CNN-based TCT cervical-cancer screening model was established through a retrospective study of multicenter TCT images. This model shows improved speed and accuracy for cervical cancer screening, and helps overcome the shortage of medical resources required for cervical cancer screening.

Background: Older people (≥60 years old) are particularly vulnerable to influenza virus infection, and vaccine is effective in reducing the disease burden in this population. However, it remains obscure whether their antibody response is lower than those of younger adults (18–60 years old). Thus, this meta-analysis was performed to compare the immunogenicity of influenza vaccines and understand their association with real-world vaccine effectiveness (VE) between these two age groups. Methods: A systematic literature search was conducted to identify relevant studies from Jan 01, 2008 to Nov 10, 2018. These are randomized controlled trials that included older adult samples, which assessed the immunogenicity of inactivated quadrivalent influenza vaccines produced in embryonated eggs. We excluded the studies focused only in children or adults. The outcomes were seroprotecton rate (SPR) and seroconversion rate (SCR). Results: Six studies were eventually included in the present meta-analysis (7,976 participants). For the SPR, the pooled risk ratio (RR) was 0.92 (95% CI: 0.90–0.94, I2 = 66%, P

Preservation of denatured dermis exerts promotive functions in wound healing and improves the appearance and function of skin. Angiogenesis is crucial for wound healing during burn injury. However, the potential molecular mechanism of angiogenesis in the recovery after burn injury remains to be elucidated. Herein, RNA chromatin immunoprecipitation (ChIP) sequencing analysis revealed upregulation of long intergenic non-coding RNA 00174 (linc00174) in the post-burn tissues. linc00174 overexpression promoted angiogenic activities of human umbilical vein endothelial cells (HUVECs) in the heat-denatured cell model, characterized by the promotion of cell proliferation, migration, and tube formation. Mechanistically, linc00174 directly bound to enhancer of zeste homolog 2 (EZH2), thus stimulating the protein level of trimethylation at lysine 27 of histone H3 (H3K27me3). Moreover, inhibition of EZH2 resulted in downregulation of ZNF24 and Runx1, as well as a decline of vascular endothelial growth factor A (VEGFA). Furthermore, EZH2 modulated epigenetic repression of ZNF24 and Runx1 through the promoter of H3K27me3. Additionally, ZNF24 and Runx1 both functioned as transcriptional inhibitors of VEGFA. Taken together, these findings uncover that linc00174 epigenetically inhibits ZNF24 and Runx1 expression through binding to EZH2, thus attenuating the suppression of VEGFA, contributing to the facilitation of angiogenesis during the recovery of heat-denatured endothelial cells.

Diffuse γ -ray emission between 10 and 1000 TeV from the Galactic plane was recently measured by the Large High Altitude Air Shower Observatory (LHAASO). These observations will help tremendously in constraining the propagation and interaction of cosmic rays in the Milky Way. Additionally, new measurements of cosmic-ray spectra reach a very high precision of up to 100 TeV energies, revealing multiple spectral structures of various species. In this work, based on up-to-date measurements of local cosmic-ray spectra and a simplified propagation setup, we confront a model prediction of diffuse γ -ray emission with measurements of diffuse γ -rays. To better constrain the low-energy part of the model, we analyze the 14.6 yr of Fermi Large Area Telescope (Fermi-LAT) data to extract the Galactic diffuse emission between 1 and 500 GeV from the same sky regions of LHAASO, after subtracting the contribution from known sources and the isotropic diffuse γ -ray background. The joint Fermi-LAT and LHAASO spectra thus cover a very wide energy range from 1 GeV to 1 PeV with small gaps from 0.5 to 10 TeV. Compared with the prediction, we find that clear excesses between several GeV and ∼60 TeV of the diffuse emission exist. Possible reasons to explain the excesses may include unresolved sources or more complicated propagation models. We illustrate that an exponential cutoff power-law component with an index of −2.40 and a cutoff energy of ∼30 TeV is able to account for such excesses.

In a national park master plan, functional zoning plays a key role in developing differentiated zoning controls that achieve multiple park construction objectives. In this study, a geographical attribute code and basic zoning elements are developed for the proposed “Ailaoshan-Wuliangshan” National Park, followed by the development of spatial multi-criteria sets and weight sets to determine the suitability of the land. Next, we use a clustering algorithm and conflict unit prioritization to allocate space for multi-target units to get the preliminary zoning schemes, and then identify stable units and unstable units through sensitivity analysis. Ultimately, the functional zoning of the National Park was determined. According to the results, the proposed “Ailaoshan-Wuliangshan” National Park can be divided into nine types of 164 landscape units; the highest land suitability values of each zone showed the traits of differentiation and aggregation in spatial distribution; there are 97 stable units and 67 unstable units; approximately 62.83% and 37.17% of the total park area can be divided into core conservation area (primary sensitive area and secondary sensitive area) and general control area (ecological activity area and ecological control area). By implementing a comprehensive assessment and decision-making process, the defined functional zones are precise and simple to recognize on the ground, and they adhere to the area proportions needed by national standards. Furthermore, the functional zoning is clustered, which avoids the fragmentation of the zoning results causing difficulties in management, and serves as a point of reference for the functional zoning approaches used in other proposed national parks in China.

The establishment of ecological networks facilitates genetic exchange among species in national parks and is an effective means of avoiding habitat fragmentation. Using the proposed “Ailaoshan-Wuliangshan” in Yunnan Province, China, as the study area, the identification of ecological source sites using the morphological spatial pattern analysis (MSPA) method, extraction of potential ecological corridors using the minimum resistance model (MCR) and construction of the ecological network of national parks were performed. Based on the gravity model, important ecological corridors were selected, and corresponding ecological network optimization strategies were presented. The results showed that (1) the core area identified by MSPA was 4440.08 km2, with a low degree of fragmentation, and is distributed in strips within the woodland land classes in the study area; (2) the establishment of an ecological network model of least cost resistance based on 10 indicators in four dimensions of land tenure, geographic factors, vegetation characteristics, and human meddling; (3) the ecological network included 13 ecological source sites, 77 potential ecological corridors, 48 important ecological corridors and 25 pedestrian pathways and extracts an optimal ecological corridor connecting with the natural reserve; and (4) the network closure degree of the constructed ecological network was (1.18), line point rate (3.08), network connectivity (1.12), and cost ratio (0.98). By using the proposed ecological network construction method, ecological patches and potential corridors can be accurately identified to ensure the integrity and connectivity of the national park while minimizing the land demand pressure of the surrounding communities, which provides some reference for the construction of other national parks’ ecological networks in China.

BackgroundThe claudin family is a group of transmembrane proteins related to tight junctions. While their involvement in cancer has been studied extensively, their relationship with the tumor immune microenvironment remains poorly understood. In this research, we focused on genes related to the prognosis of ovarian cancer and explored their relationship with the tumor immune microenvironment.MethodsThe cBioPortal for Cancer Genomics database was used to obtain the genetic variation pattern of the claudin family in ovarian cancer. The ONCOMINE and Gene Expression Profiling Interactive Analysis (GEPIA) databases were used to explore the mRNA expression of claudins in cancers. The prognostic potential of these genes was examined via the Kaplan-Meier plotter. The enrichment of immunological signatures was determined by gene set enrichment analysis (GSEA). The correlations between claudins and the tumor immune microenvironment in ovarian cancer were investigated via the Tumor Immune Estimation Resource (TIMER).ResultsClaudin genes were altered in 363 (62%) of queried patients/samples. Abnormal expression levels of claudins were observed in various cancers. Among them, CLDN3, CLDN4, CLDN6, CLDN10, CLDN15, and CLDN16 were significantly correlated with overall survival in patients with ovarian cancer. GSEA revealed that CLDN6 and CLDN10 were significantly enriched in immunological signatures of B cell, CD4 T cell, and CD8 T cell. Furthermore, CLDN6 and CLDN10 were negatively correlated and positively correlated, respectively, with immune cell infiltration in ovarian cancer. The expression levels of CLDN6 and CLDN10 were also negatively correlated and positively correlated, respectively, with various gene markers of immune cells in ovarian cancer. Thus, CLDN6 and CLDN10 may participate in immune cell infiltration in ovarian cancer, and these mechanisms may be the reason for poor prognosis.ConclusionOur study showed that CLDN6 and CLDN10 were prognostic biomarkers correlated with the immune microenvironment in ovarian cancer. These results reveal new roles for CLDN6 and CLDN10 as potential therapeutic targets in the treatment of ovarian cancer.

Abstract Neoadjuvant chemotherapy (NACT) remains an attractive alternative for controlling locally advanced cervical cancer. However, approximately 15–34% of women do not respond to induction therapy. To develop a risk stratification tool, 56 patients with stage IB‐IIB cervical cancer are included in 2 research centers from the discovery cohort. Patient‐specific somatic mutations led to NACT non‐responsiveness are identified by whole‐exome sequencing. Next, CRISPR/Cas9‐based library screenings are performed based on these genes to confirm their biological contribution to drug resistance. A 15‐gene classifier is developed by generalized linear regression analysis combined with the logistic regression model. In an independent validation cohort of 102 patients, the classifier showed good predictive ability with an area under the curve of 0.80 (95% confidence interval (CI), 0.69–0.91). Furthermore, the 15‐gene classifier is significantly associated with patient responsiveness to NACT in both univariate (odds ratio, 10.8; 95% CI, 3.55–32.86; p = 2.8 × 10−5) and multivariate analysis (odds ratio, 17.34; 95% CI, 4.04–74.40; p = 1.23 × 10−4) in the validation set. In conclusion, the 15‐gene classifier can accurately predict the clinical response to NACT before treatment, representing a promising approach for guiding the selection of appropriate treatment strategies for locally advanced cervical cancer.

Abstract Among all malignant tumors that threaten human health, virus-related tumors account for a large proportion. The treatment of these tumors is still an urgent problem to be resolved. The immune system is the “guard” of the human body, resisting the invasion of foreign substances such as viruses. Studies have shown that immunotherapy has clinical significance in the treatment of a variety of tumors. In particular, the emergence of immune checkpoint inhibitors (ICIs) in recent years has opened a new door to cancer therapy. Considering the potential role of ICIs in the treatment of virus-related cancers, we focused on their therapeutic effect in virus-associated cancers and explored whether the therapeutic effect in virus-associated cancers was related to virus infection status. Although there is no clear statistical significance indicates that ICIs are more effective in virus-associated cancers than non-virus infections, the efficacy of checkpoint inhibitors in the treatment of virus-related cancers is promising. We believe that this research provides a good direction for the implementation of individualized precision medicine.

Background/Aims: To demonstrate the function of uncoupling protein 2 (UCP2) in the regulation of human spermatozoa motility. Methods: Semen samples were collected from donors with either normal spermatozoa motility (normospermia [NS]) or poor spermatozoa motility (asthenospermia [AS]). UCP2 protein in spermatozoawas quantified by Western blotting. The level of mitochondrial reactive oxygen species (mROS) was evaluated by MitoSOX Red. The activity of mitochondrial membrane potential (MMP) in spermatozoa was evaluated by a JC-1 assay and the ATP level was monitored by a luciferin-luciferase assay. Results: UCP2 was expressed in both NS and AS groups, with the former exhibiting a higher level than the latter. Immunofluorescence analysis shows that UCP2 is mainly located at the mid-region of human spermatozoa. The inhibition of UCP2 by a highly selective inhibitor, Genipin, results in not only impaired spermatozoa mobility (P

RAD54B, a human paralog of RAD54 L, belongs to the SW12/SNF2 superfamily of helicases. Up to now, few studies have reported the role of RAD54B in breast cancer (BC). In this study, we reported that RAD54B was amplified and the expression of RAD54B was elevated in BC, as identified through a bioinformatics analysis of open databases (cBioPortal and Oncomine) and an immunohistochemical analysis of a tissue microarray (TMA). Analysis of Kaplan-Meier plotter (KM plotter) showed that high RAD54B transcription activity was critically linked with poor prognosis of BC patients, in particular those with the luminal A subtype. Gene set enrichment analysis (GSEA) performed on dataset GSE1456 demonstrated that gene expression signatures associated with survival, proliferation, cell cycle, apoptosis and p53 signaling were crucially enriched, when the level of RAD54B was elevated. Further, gain-of-function studies were conducted on luminal A BC cell lines MCF-7 and ZR-751 to validate these findings. Consistently, RAD54B knockdown suppressed cell proliferation in vitro, as well as delayed tumor growth in vivo. Collectively, our results address the biological role of RAD54B in the neoplastic process of luminal A BC. Bioinformatics analysis, additionally, indicates RAD54B with a predictive value for this BC subtype.

Pseudomonas aeruginosa -derived pigment pyocyanin (PCN) has been proved to induce cell apoptosis mediated by the generation of reactive oxygen species (ROS), which has been studied mainly in epithelial cells and neutrophils. However, we previously found that the PCN-producing strain PA14 induces cell apoptosis in human NK cell line NK92 more effectively than in PCN-deficient strain PA14-phZ1/2 via a yet undetermined mechanism. In the current study, we found that PCN-induced NK92 cell apoptosis occurs through mitochondrial damage despite inhibiting intracellular ROS generation. Intracellular Ca 2+ ([Ca 2+ ] i ) and Bcl-2 family proteins act as important “priming signals” for apoptosis. PCN treatment increased [Ca 2+ ] i in NK92 cells more than twofold after 2 h stimulation, whereas the Ca 2+ -chelating agent ethylene glycol tetra-acetic acid (EGTA) inhibited apoptosis. PCN triggered the activation of Bim, Bid, Bik, Bak, and phospho-Bad in NK92 cells in a concentration-dependent manner, but these pro-apoptotic Bcl-2 family proteins were not inhibited by EGTA. In this study, we describe the function of PCN in NK92 cells and identify mitochondrial damage as the mechanism underlying the apoptosis. [Ca 2+ ] i and pro-apoptotic Bcl-2 family proteins are novel targets for PCN-induced apoptosis. Clarification of the cytotoxic diversity of PCN provides a new therapeutic target for defense from P. aeruginosa -induced immune cell damage.

Our previous study has suggested that downregulated microRNA (miR)-29a in denatured dermis might be involved in burn wound healing. However, the exact role of miR-29a in healing of burn injury still remains unclear. Here, we found that expression of miR-29a was notably upregulated in denatured dermis tissues and skin fibroblast cells after thermal injury, and thereafter gradually downregulated compared with control group. By contrast, the expression of collagen, type I, alpha 2 (COL1A2) and vascular endothelial growth factor (VEGF-A) were first reduced and subsequently upregulated in denatured dermis tissues and skin fibroblast cells after thermal injury. We further identified COL1A2 as a novel target of miR-29a, which is involved in type I collagen synthesis, and showed that miR-29a negatively regulated the expression level of COL1A2 in skin fibroblast cells. In addition, VEGF-A, another target gene of miR-29a, was also negatively mediated by miR-29a in skin fibroblast cells. Inhibition of miR-29a expression significantly promoted the proliferation and migration of skin fibroblast cells after thermal injury, and knockdown of COL1A2 and VEGF-A reversed the effects of miR-29a on the proliferation and migration of skin fibroblast cells. Furthermore, we found that Notch2/Jagged2 signaling was involved in miR-29a response to burn wound healing. Our findings suggest that downregulated miR-29a in denatured dermis may help burn wound healing in the later phase, probably via upregulation of COL1A2 and VEGF-A expression, which can further enhance type I collagen synthesis and angiogenesis.

Invasive fungal infection is one of the major complication of severe burns which can induce local or systemic inflammatory response and cause serious substantial damage to the patient. The incidence of fungal infection for burn victims is increasing dramatically during recent years. This guideline, organized by Chinese Society of Burn Surgeons, aims to standardize the diagnosis, prevention and treatment of burn invasive fungal infection. It can be used as one of the tools for treatment of major burn patients.

Skin grafting with the preservation of denatured dermis is a novel strategy for the treatment of burn-injured skin. Denatured dermis has the ability to restore to the morphology and function of normal skin, but the underlying molecular mechanism is elusive. MicroRNAs (miRNA) are small noncoding RNAs and regulate normal physiology as well as disease development. In this study, we assessed the potential role of miRNA-23b (miR-23b) in the regulation of cell proliferation and migration of heat-denatured fibroblasts and identified the underlying mechanism.The expression of miR-23b in denatured dermis and heat-denatured fibroblasts was detected by quantitative real-time polymerase chain reaction (RT-PCR). The effects of miR-23b on cell proliferation and migration of heat-denatured fibroblasts were assessed by transient transfection of miR-23b mimics and inhibitor. The target gene of miR-23b and the downstream pathway were further investigated.miR-23b was downregulated in denatured dermis and heat-denatured fibroblasts. Downregulation of miR-23b dramatically promoted the proliferation and migration of heat-denatured fibroblasts. Subsequent analyses demonstrated that Smad3 was a direct and functional target of miR-23b in heat-denatured fibroblasts, which was validated by the dual luciferase reporter assay. Moreover, immunohistochemistry analysis showed that denatured dermis from rats displayed enhanced staining of Smad3. In addition, miR-23b modulated denatured dermis by activating the Notch1 and TGF-β signaling pathways.Our findings suggest that downregulation of miR-23b contributes to the recovery of denatured dermis, which may be valuable for treatment of skin burns.

BackgroundT cell immunoglobulin mucin-3 (Tim-3) has been identified as a negative regulator of anti-tumor immunity. Recent studies highlight the important role of Tim-3 in the CD8(+) T cell exhaustion that takes place in both human and animal cancer models. However, the nature of Tim-3 expression in the tumor cell and the mechanism by which it inhibits anti-tumor immunity are unclear. This present study aims to determine Tim-3 is expressed in cervical cancer cells and to evaluate the role of Tim-3 in cervical cancer progression.MethodologyA total of 85 cervical tissue specimens including 43 human cervical cancer, 22 cervical intraepithelial neoplasia (CIN) and 20 chronic cervicitis were involved. Tim-3 expression in tumor cells was detected and was found to correlate with clinicopathological parameters. Meanwhile, expression of Tim-3 was assessed by RT-PCR, Western Blot and confocal microscopy in cervical cancer cell lines, HeLa and SiHa. The migration and invasion potential of Hela cells was evaluated after inhibiting Tim-3 expression by ADV-antisense Tim-3.ConclusionsWe found that Tim-3 was expressed at a higher level in the clinical cervical cancer cells compared to the CIN and chronic cervicitis controls. We supported this finding by confirming the presence of Tim-3 mRNA and protein in the cervical cell lines. Tim-3 expression in tumor cells correlated with clinicopathological parameters. Patients with high expression of Tim-3 had a significant metastatic potential, advanced cancer grades and shorter overall survival than those with lower expression. Multivariate analysis showed that Tim-3 expression was an independent factor for predicting the prognosis of cervical cancer. Significantly, down-regulating the expression of Tim-3 protein inhibited migration and invasion of Hela cells. Our study suggests that the expression of Tim-3 in tumor cells may be an independent prognostic factor for patients with cervical cancer. Moreover, Tim-3 expression may promote metastatic potential in cervical cancers.

With the deep integration of modern information technology and auxiliary training, virtual reality technology represented by human-computer interaction has gradually received widespread attention. In this paper, virtual reality technology is applied to the physical fitness training of college engineering students to assist teaching, so that learners in the virtual sports situation with the help of VR sensing equipment to complete a variety of sensory real-time interaction, combined with the goal of stimulating feedback mechanism to improve students' self-efficacy and interest in learning, and use practice to explore the teaching effect and potential value of the achieved not only to promote the diversified development of sports skills learning, but also to provide a reference for the application of virtual reality technology in sports. It can not only promote the diversified development of motor skills learning, but also provide reference for the application of virtual reality technology in sports. The application of virtual reality technology in the physical fitness training and teaching of college engineering students can be used as a means to stimulate the effect of the potential learning ability of the learners, in the awakening of interest in the sports situation at the same time to promote the improvement of sports skills, while the traditional classroom learners in the emotional attitude did not have a positive effect on the promotion. Secondly, it is difficult to form an effective stimulating effect in physical quality by different teaching methods. The experimental results show that the application of using virtual reality technology to optimise engineering students' physical fitness training is significant and proves the effectiveness of the proposed method. [ABSTRACT FROM AUTHOR]