A conserved methyltransferase active site residue of Zika virus NS5 is required for the restriction of STING activation and interferon expression.

Zika virus (ZIKV) is a re-emerging RNA virus and causes major public health events due to its link to severe neurological complications in foetuses and neonates. The cGAS-STING signalling pathway regulates innate immunity and plays an important role in the invasion of DNA and RNA viruses. This study reveals a distinct mechanism by which ZIKV restricts the cGAS-STING signalling to repress IFN-ß expression. ZIKV attenuates IFN-ß expression induced by DNA viruses (herpes simplex virus type 1, HSV-1) or two double-stranded DNAs (dsDNA90 and HSV120) in mouse embryonic fibroblasts (MEFs). Notably, ZIKV NS5, the viral RNA-dependent RNA polymerase, was responsible for the repression of IFN-ß. NS5 interacts with STING in the cytoplasm, suppresses IRF3 phosphorylation and nucleus localization and promotes the cleavage of STING K48-linked polyubiquitination. Furthermore, the NS5 methyltransferase (MTase) domain interacts with STING to restrict STING-induced IFN-ß expression. Interestingly, point mutation analyses of conserved methyltransferase active site residue D146 indicate that it is critical for repressing IFN-ß expression induced by STING stimulation in cGAS-STING signalling.

A transcriptome based molecular classification scheme for cholangiocarcinoma and subtype-derived prognostic biomarker.

Previous studies on the molecular classification of cholangiocarcinoma (CCA) focused on certain anatomical sites, and disregarded tissue contamination biases in transcriptomic profiles. We aim to provide universal molecular classification scheme and prognostic biomarker of CCAs across anatomical locations. Comprehensive bioinformatics analysis is performed on transcriptomic data from 438 CCA cases across various anatomical locations. After excluding CCA tumors showing normal tissue expression patterns, we identify two universal molecular subtypes across anatomical subtypes, explore the molecular, clinical, and microenvironmental features of each class. Subsequently, a 30-gene classifier and a biomarker (called "CORE-37") are developed to predict the molecular subtype of CCA and prognosis, respectively. Two subtypes display distinct molecular characteristics and survival outcomes. Key findings are validated in external cohorts regardless of the stage and anatomical location. Our study provides a CCA classification scheme that complements the conventional anatomy-based classification and presents a promising prognostic biomarker for clinical application.

Molecular profiles of different PD-L1 expression in patients with esophageal squamous cell carcinoma.

BACKGROUND: PD-1/PD-L1 inhibitors are approved treatments for patients with esophageal squamous cell carcinoma (ESCC). The present investigation aspired to explore the interrelation between molecular phenotype and PD-L1 expression in ESCC. METHODS: PD-L1 testing and targeted next-generation sequencing (NGS) were performed on tumoral tissues from 139 ESCC patients. Tumor-infiltrating lymphocytes (TILs) were scrutinized using a tyramide signal amplification system combined with immunohistochemistry. RESULTS: Among enrolled patients, 36.7% displayed high PD-L1 expression (combined positive score [CPS] ≥10). BRCA1 and NF1 gene mutations were significantly associated with high PD-L1 expression (p < .05) while TGFß pathway alterations were linked to low PD-L1 expression (p = .02). High copy number instability (CNI) and copy number alterations (CNA) were correlated with low PD-L1 expression. Patients with CDKN2A deletion exhibited higher PD-L1 expression. Varying types of TILs were observed across different PD-L1 expression groups. The ratio of CD8+PD-L1+ T cells and CD8+PD-1+ T cells to CD8+ T cells remained comparable in both tumoral and stromal regions, but the ratio of CD68+PD-L1+ macrophages to CD68+ macrophages was higher than the ratio of CD68+PD-1+ macrophages to CD68+ macrophages. CPS was significantly correlated with PD-L1+ lymphocytes and CD68+ macrophages in the tumoral region. CD8+ T cell infiltration was positively correlated with PD-1+ cells in both tumoral and stromal regions. CONCLUSION: In this study, we presented the prevalence rates of PD-L1 expression in Chinese ESCC patients. The association of genetic profiles with PD-L1 expression levels also provide the clue that genomic phenotype may interact with the immunologic phenotype in ESCC.

The in vitro and in vivo antiviral effects of aloperine against Zika virus infection.

Zika virus (ZIKV) infection poses a significant threat to global public health and is associated with microcephaly. There are no approved ZIKV-specific vaccines or drugs for the clinical treatment of the infection. Currently, there are no approved ZIKV-specific vaccines or drugs for the clinical treatment of the infection. In this study, we investigated the antiviral potential of aloperine, a quinolizidine alkaloid, against ZIKV infection in vivo and in vitro. Our results demonstrate that aloperine effectively inhibits ZIKV infection in vitro, with a low nanomolar half maximal effective concentration (EC50 ). Specifically, aloperine strongly protected cells from ZIKV multiplication, as indicated by decreased expression of viral proteins and virus titer. Our further investigations using the time-of-drug-addition assay, binding, entry, and replication assays, detection of ZIKV strand-specific RNA, the cellular thermal shift assay, and molecular docking revealed that aloperine significantly inhibits the replication stage of the ZIKV life cycle by targeting the domain RNA-dependent RNA polymerase (RDRP) of ZIKV NS5 protein. Additionally, aloperine reduced viremia in mice and effectively lowered the mortality rate in infected mice. These findings highlight the potency of aloperine and its ability to target ZIKV infection, suggesting its potential as a promising antiviral drug against ZIKV.

Single-cell RNA sequencing highlights the role of PVR/PVRL2 in the immunosuppressive tumour microenvironment in hepatocellular carcinoma.

Introduction: The conflict between cancer cells and the host immune system shapes the immune tumour microenvironment (TME) in hepatocellular carcinoma (HCC). A deep understanding of the heterogeneity and intercellular communication network in the TME of HCC will provide promising strategies to orchestrate the immune system to target and eradicate cancers. Methods: Here, we performed single-cell RNA sequencing (scRNA-seq) and computational analysis of 35786 unselected single cells from 3 human HCC tumour and 3 matched adjacent samples to elucidate the heterogeneity and intercellular communication network of the TME. The specific lysis of HCC cell lines was examined in vitro using cytotoxicity assays. Granzyme B concentration in supernatants of cytotoxicity assays was measured by ELISA. Results: We found that VCAN+ tumour-associated macrophages (TAMs) might undergo M2-like polarization and differentiate in the tumour region. Regulatory dendritic cells (DCs) exhibited immune regulatory and tolerogenic phenotypes in the TME. Furthermore, we observed intensive potential intercellular crosstalk among C1QC+ TAMs, regulatory DCs, regulator T (Treg) cells, and exhausted CD8+ T cells that fostered an immunosuppressive niche in the HCC TME. Moreover, we identified that the TIGIT-PVR/PVRL2 axis provides a prominent coinhibitory signal in the immunosuppressive TME. In vitro, antibody blockade of PVR or PVRL2 on HCC cell lines or TIGIT blockade on immune cells increased immune cell-mediated lysis of tumour cell. This enhanced immune response is paralleled by the increased secretion of Granzyme B by immune cells. Discussion: Collectively, our study revealed the functional state, clinical significance, and intercellular communication of immunosuppressive cells in HCC at single-cell resolution. Moreover, PVR/PVRL2, interact with TIGIT act as prominent coinhibitory signals and might represent a promising, efficacious immunotherapy strategy in HCC.

Universal Detection and Imaging for Multiple Targets by Coupling Target-Primed Nicking-Enhanced Rolling Circle Amplification with Self-Powered DNAzyme Walker.

Although promising in monitoring low-abundance analytes, most of the DNAzyme walker is only responsive to a specific target. Herein, a universal, ready-to-use platform is developed by coupling nicking-enhanced rolling circle amplification and a self-powered DNAzyme walker (NERSD). It addressed the issues that DNAzyme strands need to be specifically designed for different biosensing system, allowing highly sensitive analysis of various targets with the same DNAzyme walker components. It is also specific owing to target-dependent ligation of the padlock probe and precise cleavage of a substrate by a DNAzyme strand. As typically demonstrated, the strategy has an equivalent capacity with the qRT-PCR kit in distinguishing plasma miR-21 levels of breast cancer patients from normal subjects and is able to differentiate intracellular miR-21 and ATP levels by confocal imaging. The approach characteristic of programmability, flexibility, and generality indicated the potential in all kinds of biosensing and imaging platform.

Facile construction of photocatalytic cellulose-based sponge with stable flotation properties as efficient and recyclable photocatalysts for sewage treatment.

Dispersion and recycling of powdered nano-photocatalysts for water purification is still not an easy task. The self-supporting and floating photocatalytic cellulose-based sponges ware conveniently prepared by anchoring BiOX nanosheet arrays on cellulose-based sponge's surface. The introduction of sodium alginate into the cellulose-based sponge significantly enhanced the electrostatic adsorption of bismuth oxygen ions and promoted the formation of bismuth oxyhalide (BiOX) crystal nuclei. Among the photocatalytic cellulose-based sponges, the sponge (BiOBr-SA/CNF) modified with bismuth oxybromide displayed excellent photocatalytic ability for photodegrading 96.1 % rhodamine B within 90 min under 300 W Xe lamp irradiation (λ > 400 nm). The loading of bismuth oxybromide on cellulose-based sponge's surface improves the flotation stability of the cellulose-based sponge. Benefiting from excellent load fastness of bismuth oxybromide nanosheet and flotation stability of BiOBr-SA/CNF sponge, after 5 cycles of recycling, the photodegradation rates of BiOBr-SA/CNF sponge to rhodamine B remained above 90.2 % (90 min), and it also has excellent photocatalytic degradation effect on methyl orange and herbicide isoproteron. This work may provide a convenient and efficient method to construct self-supporting and floating photocatalytic sponges using cellulose based materials as substrates for sewage treatment.

Transcriptomic reveals the ferroptosis features of host response in a mouse model of Zika virus infection.

Zika virus (ZIKV) is a neurotropic flavivirus. The outbreak of ZIKV in 2016 created a global health emergency. However, the underlying pathogenic mechanisms remain elusive. We investigated the host response features of in vivo replication in a mouse model of ZIKV infection, by performing a series of transcriptomic and bioinformatic analyses of ZIKV and mock-infected brain tissue. Tissue damage, inflammatory cells infiltration and high viral replication were observed in the brain tissue of ZIKV infected mice. RNA-Seq of the brain indicated the activation of ferroptosis pathways. Enrichment analysis of ferroptosis regulators revealed their involvement in pathways such as mineral absorption, fatty acid biosynthesis, fatty acid degradation, PPAR signaling pathway, peroxidase, and adipokinesine signalling pathway. We then identified 12 interacted hub ferroptosis regulators (CYBB, HMOX1, CP, SAT1, TF, SLC39A14, FTL, LPCAT3, FTH1, SLC3A2, TP53, and SLC40A1) that were related to the differential expression of CD8+ T cells, microglia and monocytes. CYBB, HMOX1, SALT, and SLAC40A1 were selected as potential biomarkers of ZIKV infection. Finally, we validated our results using RT-qPCR and outside available datasets. For the first time, we proposed a possible mechanism of ferroptosis in brain tissue infected by ZIKV in mice and identified the four key ferroptosis regulators.

Scalable Fabrication of Highly Breathable Cotton Textiles with Stable Fluorescent, Antibacterial, Hydrophobic, and UV-Blocking Performance.

Multifunctional cotton textiles that are highly breathable are desirable in a broad range of applications. However, it is still a big challenge to scale up production of such multifunctional cotton textiles. Herein, we developed a simple, scalable, and benign strategy to fabricate highly breathable multifunctional cotton textiles via mild surface modification. The 1,4-dihydropyridine (DHP) ring and gentamycin sulfate (GS) molecules were firmly attached to the cellulose chains under room temperature via a one-pot method. The resulting modified cotton textile showed integrated performances with bright fluorescence, good antibacterial behavior, hydrophobic behavior (contact angle of 134°), and UV-blocking (UPF being up to 69.2), which are very stable toward washing and various solvents. There is no obvious change in the whiteness, thermal stability, and mechanical performance of cotton fabrics after the surface modification. What's more, the air permeability of the modified cotton fabric was up to 31.3 (cm3/cm2)/s. This study not only focuses on the materials design and large-scale fabrication but also provides stable and multifunctional cotton textiles with broad application prospects for many fields.

Levels of circulating mast cell progenitors and tumour­infiltrating mast cells in patients with colorectal cancer.

The role of mast cells in colorectal cancer (CRC) has been an area of intense interest. Mast cell density is closely related to CRC development and prognosis. The identification of mast cell progenitors (MCps) in peripheral blood provides an opportunity to explore the frequency and distribution of mast cells in the circulation and tumour microenvironment of patients with CRC at different disease stages. The aim of the presents study was to investigate the changes of MCps and mast cells in CRC. Flow cytometry was used to measure the circulating frequency of MCps in 37 patients with CRC and 12 healthy control (HC) patients, and the frequency of mast cells in tissue from 15 patients with CRC and 7 patients with haemorrhoids. In the present study, lower levels of circulating MCps in patients with CRC were found, which was significantly related to CRC development. After surgery, the frequency of circulating MCps was significantly increased. However, the frequency of mast cells in tumour tissues was lower than that in adjacent normal tissues and compared with HC tissues and was not associated with CRC progression.

Research progress and challenges to coronavirus vaccine development.

Coronaviruses (CoVs) are nonsegmented, single-stranded, positive-sense RNA viruses highly pathogenic to humans. Some CoVs are known to cause respiratory and intestinal diseases, posing a threat to the global public health. Against this backdrop, it is of critical importance to develop safe and effective vaccines against these CoVs. This review discusses human vaccine candidates in any stage of development and explores the viral characteristics, molecular epidemiology, and immunology associated with CoV vaccine development. At present, there are many obstacles and challenges to vaccine research and development, including the lack of knowledge about virus transmission, pathogenesis, and immune response, absence of the most appropriate animal models.

A Randomized Controlled Comparison of Epidural Analgesia Onset Time and Adverse Reactions During Labor With Different Dose Combinations of Bupivacaine and Sufentanil.

OBJECTIVES: The purpose was to compare the effects of 3 different dose combinations of bupivacaine and sufentanil on the onset of analgesia and the occurrence of side effects. MATERIALS AND METHODS: One hundred sixty-nine pregnant women were randomly assigned to 3 groups: the B1S5 group received 0.1% bupivacaine+5 µg sufentanil in 15 mL; the B125S5 group received 0.125% bupivacaine+5 µg sufentanil in 15 mL; and the B1S10 group received 0.1% bupivacaine+10 µg sufentanil in 15 mL. The primary outcome was the analgesic onset time, and the secondary outcomes were mode of delivery, patient satisfaction, maternal and neonatal side effects (pruritus, hypotension, sedation, motor block, decreased fetal heart rate, fever, and interference with breastfeeding). RESULTS: The median (inter-quartile range) time to achieve effective analgesia was significantly faster in the B125S5 group than in the B1S5 group (10 [11-14 {4-30}] min vs. 15 [17-20 {5-30}] min, P<0.001). There was no significant difference in the analgesia onset time between the B1S10 and B125S5 groups (10 [11-14 {4-30}] min vs. 12 [13-15 {3-30}] min, P=0.202). Pruritus, hypotension, motor block, maternal satisfaction, delivery mode, decreased fetal heart rate, total bupivacaine dose and breastfeeding scores were not significantly different among the 3 groups except the sufentanil dosage and incidence of mild drowsiness and fever (the B1S10 group had significantly higher fever than the other groups). DISCUSSION: The B125S5 combination may be superior to the B1S5 and B1S10 combinations as an initial dose for epidural analgesia to achieve rapid effective analgesia with minimal side effects.

Coronavirus infections and immune responses.

Coronaviruses (CoVs) are by far the largest group of known positive-sense RNA viruses having an extensive range of natural hosts. In the past few decades, newly evolved Coronaviruses have posed a global threat to public health. The immune response is essential to control and eliminate CoV infections, however, maladjusted immune responses may result in immunopathology and impaired pulmonary gas exchange. Gaining a deeper understanding of the interaction between Coronaviruses and the innate immune systems of the hosts may shed light on the development and persistence of inflammation in the lungs and hopefully can reduce the risk of lung inflammation caused by CoVs. In this review, we provide an update on CoV infections and relevant diseases, particularly the host defense against CoV-induced inflammation of lung tissue, as well as the role of the innate immune system in the pathogenesis and clinical treatment.

Chitosan-bound carboxymethylated cotton fabric and its application as wound dressing.

Cotton fabric (CF) is commonly used in wound treatment, however, its hemostatic efficiency is far from sufficient. In this study, modified cotton fabric (MCF-0.39) was obtained by a carboxymethylation process, which endowed MCF-0.39 with good swelling ability and water absorption capacity. Chitosan (CHI) was bound to MCF-0.39 by the binder sodium carboxymethyl cellulose (NaCMC) via flat-screen printing technique to prepare the hybrid hemostatic material (CHI-MCF-0.39). The blood clotting index (BCI) of CHI-MCF-0.39 was 3.15-fold lower than that of CF, demonstrating the good clotting ability of the material. In rat liver injury and femoral artery animal model, the groups using CHI-MCF-0.39 had less hemostasis time and blood loss compared with those groups using CF. All the above results indicate that the prepared CHI-MCF-0.39 has promising future applications as effective hemostatic material in trauma treatment.

Determination of the optimal reaction conditions for the preparation of highly substituted carboxymethyl Cassia tora gum.

Highly substituted carboxymethyl Cassia tora gum (CM-CTG) was prepared from CTG by treatment with monochloroacetic acid (MCA) in ethanolic aqueous solutions after alkalization under different reaction conditions. The influence of the etherification temperature, alkalization and etherification times, molar ratio of sodium hydroxide to MCA (nNaOH/nMCA), theoretical degree of substitution (DSt), and weight percentage of water (WH2O%) in the solution on the degree of substitution (DS) and reaction efficiency (RE) of the reaction were investigated. Optimum preparation conditions for a CM-CTG with DS of 1.05 are: etherification temperature, 60°C; alkalization time, 60min; etherification time, 180min; nNaOH/nMCA, 2.1; DSt, 1.4; and WH2O%, 20%. Fourier-transform infrared analysis of the products indicated that carboxymethylation was successful. Rheological studies show that all the CM-CTG pastes are pseudoplastic fluids, and the shear sensitivity varies with DS. The degree of crystallinity of CM-CTG decreases with increasing DS, as shown by X-ray diffraction measurements.

Novamin infusion: a new method to cure postoperative shivering with hypothermia.

BACKGROUND: Postoperative shivering is a frequent complication of surgery in developing countries and there is no satisfying method to treat it, let alone to cure it. We studied whether intravenous amino acid (AA) infusion can cure postoperative shivering in the postanesthesia care unit. METHODS: Sixty postanesthesia care unit patients with shivering grade 2 or higher and tympanic temperature <36°C received randomly either infusion of Novamin 18 AAs (2 mL/kg/h), pethidine (0.5 mg/kg), or tramadol (1 mg/kg). Tympanic temperature, shivering grade, and thermal comfort were assessed every 5 min for 60 min. Blood glucose and lactic acid concentrations were measured before and after treatment. Postoperative nausea and vomiting were also recorded. RESULTS: Shivering stopped within 5 min in the pethidine and tramadol groups versus 90% stopped within 15 min in AA group. There were five cases of reshivering in the tramadol group versus none in the AA or pethidine groups. Tympanic temperature increased slowly in all patients but increased significantly faster in the AA group. Thermal comfort improved significantly faster in the AA group versus the other two groups, thermal comfort was significantly higher in the tramadol versus the pethidine group ≥35 min. Blood glucose concentration in AA group increased to 135.18 ± 9.18 mg/dL. There were some cases of nausea and vomiting in pethidine and tramadol groups but none in the AA group. CONCLUSION: Novamin infusion can stop postoperative shivering and alleviates hypothermia and improves thermal comfort more effectively than tramadol and pethidine with less nausea and vomiting and causes a clinically acceptable blood glucose increase with no reshivering episodes.

Effects of different loading doses of dexmedetomidine on bispectral index under stepwise propofol target-controlled infusion.

BACKGROUND AND PURPOSE: Stepwise propofol target-controlled infusion (TCI) can achieve a less disturbed condition of hemodynamics and respiration. Its combination with dexmedetomidine may have some advantages for patients. We studied the effects of different loading doses of dexmedetomidine on the bispectral index (BIS) under stepwise propofol TCI. METHODS: Forty patients were randomly assigned into groups D(1.0), D(0.5), D(0.25) and D(0), in which dexmedetomidine at 1.0, 0.5, 0.25 or 0 µg•kg(-1) was infused over 10 min followed by 0.5 µg•kg(-1)•h(-1) and stepwise propofol TCI, which was administered with target effect site concentration (Ce) at 0.5 µg•ml(-1), and increased until 2.5 µg•ml(-1) by 1.0 µg•ml(-1) after 5 min reaching target Ce. BIS, heart rate, MAP, pulse oxygen saturation, RR and end-tidal carbon dioxide pressure were recorded before loading dose (T(0)), at 5 min (T(5 min)) and 10 min (T(10 min)) after starting infusion, after 5 min reaching Ce of 0.5, 1.5 and 2.5 µg•ml(-1) (T(p0.5), T(p1.5) and T(p2.5)). RESULTS: BIS values in group D(1.0) were significantly lower compared with those in group D(0) since T(10 min) and those in groups D(0.5) and D(0.25) since T(p0.5). In group D(1.0), heart rate decreased significantly at T(5 min) and T(10 min), heart rate at T(10 min) was significantly lower compared with that in group D(0). MAP remained stable during the loading dose infusion and decreased to some degree after propofol infusion in all groups. Changes in pulse oxygen saturation, RR and end-tidal carbon dioxide pressurewere similar among the groups without respiration depression. CONCLUSION: A loading dose of dexmedetomidine of 1.0 µg•kg(-1), not 0.5 µg•kg(-1) or less, over 10 min followed by 0.5 µg•kg(-1)•h(-1) can definitely decrease the BIS under stepwise propofol TCI with clinically stable blood pressure and without respiration depression, while attention should be paid to decreased heart rate.

The design, synthesis, and biological evaluation of PIM kinase inhibitors.

A series of substituted benzofuropyrimidinones with pan-PIM activities and excellent selectivity against a panel of diverse kinases is described. Initial exploration identified aryl benzofuropyrimidinones that were potent, but had cell permeability limitation. Using X-ray crystal structures of the bound PIM-1 complexes with 3, 5m, and 6d, we were able to guide the SAR and identify the alkyl benzofuropyrimidinone (6l) with good PIM potencies, permeability, and oral exposure.

Discovery of XL413, a potent and selective CDC7 inhibitor.

CDC7 is a serine/threonine kinase that has been shown to be required for the initiation and maintenance of DNA replication. Up-regulation of CDC7 is detected in multiple tumor cell lines, with inhibition of CDC7 resulting in cell cycle arrest. In this paper, we disclose the discovery of a potent and selective CDC7 inhibitor, XL413 (14), which was advanced into Phase 1 clinical trials. Starting from advanced lead 3, described in a preceding communication, we optimized the CDC7 potency and selectivity to demonstrate in vitro CDC7 dependent cell cycle arrest and in vivo tumor growth inhibition in a Colo-205 xenograft model.