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Osteosarcoma, which primarily affects children and adolescents, is a highly malignant bone tumor with high rates of disability and mortality. Therefore, the exploration of biological markers related to its occurrence, development, and prognosis is crucial. Genome-wide association studies have revealed the vital role of genetic polymorphisms in the pathogenesis of osteosarcoma. This study aims to provide new insights into reliable biomarkers of osteosarcoma through the analysis of the functional single-nucleotide polymorphisms of tumor-related genes.

This study compared the feasibility of circulating fluidized bed boiler fly ash (CFBFA), pulverized coal furnace fly ash (PCFA), and coal gasification slag (CGS) for the preparation of geopolymers by investigating the Si and Al dissolution reaction of the raw materials, product structure, and the development of the mechanical properties of the geopolymers. The results indicated that the reaction mechanism for preparing geopolymers were the same for the three kinds of coal based ash, i.e., the depolymerization and repolymerization of Si-O and Al-O bonds in raw materials. The loose and porous microstructure of CFBFA resulted in a high dissolution ratio of amorphous Si and Al, and its self-hardening properties generated ettringite crystals. This resulted in a high initial strength of the CFBFA based geopolymer (CFBFAG). The active Si and Al in PCFA and CGS were released slowly during the curing process, which was conducive to an enhanced compressive strength of the geopolymer in the later stage. The compressive strengths of the CGS based geopolymer (CGSG) after curing for 90 days reached 52.1 MPa, which was 139 % and 11.4 % higher than that of CFBFAG and PCFAG for the same curing time respectively. A microstructure analysis showed that the CGSG had a denser microstructure and more ordered geopolymer gel network than the other geoploymers. Compared with the CFBFA and PCFA, CGS was found to have a greater application potential in the preparation of geopolymers.

ABSTRACTIdentifying tumor-relevant T cell subsets in the peripheral blood (PB) has become a potential strategy for cancer treatment. However, the subset of PB that could be used to treat cancer remains poorly defined. Here, we found that the CX3CR1+ T cell subset in the blood of patients with lung cancer exhibited effector properties and had a higher TCR matching ratio with tumor-infiltrating lymphocytes (TILs) compared to CX3CR1− T cells, as determined by paired single-cell RNA and TCR sequencing. Meanwhile, the anti-tumor activities, effector cytokine production, and mitochondrial function were enhanced in CX3CR1+ T cells both in vitro and in vivo. However, in the co-culture system of H322 cells with T cells, the percentages of apoptotic cells and Fas were substantially higher in CX3CR1+ T cells than those in CX3CR1− T cells. Fas-mediated apoptosis was rescued by treatment with an anti-PD-1 antibody. Accordingly, the combination of adoptive transfer of CX3CR1+ T cells and anti-PD-1 treatment considerably decreased Fas expression and improved the survival of lung xenograft mice. Moreover, an increased frequency of CX3CR1+ T cells in the PB correlated with a better response and prolonged survival of patients with lung cancer who received anti-PD-1 therapy. These findings indicate the promising potential of adoptive transfer of peripheral CX3CR1+ T cells as an individual cancer immunotherapy.

Oxygen minimum zones (OMZs) in the ocean are areas with dissolved oxygen (DO) concentrations below critical thresholds that impact marine ecosystems and biogeochemical cycling. In the northern Indian Ocean (NIO), OMZs exhibit a tendency to expand in mesopelagic waters and contribute significantly to global nitrogen loss and climate change. However, the microbial drivers of OMZ expansion in the NIO remain understudied. Here, we characterized bacterial communities across DO gradients in the NIO using high-throughput 16S rRNA gene sequencing. We found that Marinimicrobia, Chloroflexi, and the SAR324 clade were enriched in both oxygen-deficient and low oxygen mesopelagic waters. Furthermore, Marinimicrobia, Chloroflexi, and the SAR324 clade exhibited a significant negative correlation with DO (P < 0.01), suggesting that they were well-adapted to the oxygen-deficient OMZ habitat. Functional predictions revealed heightened nitrogen metabolism in OMZs, particularly nitrate reduction, suggesting its pivotal role in nitrogen loss. These findings underscore the importance of microbial communities in driving OMZ expansion in the NIO and highlight their implications in global biogeochemical cycles and climate change.

In this study, an optimized composite was prepared based on chemically modified corn straw, chitosan, and poly(lactic acid) using Response Surface Methodology (RSM). The composite was produced by screw extruding and hot pressing. The Box Behnken Design (BBD) software was used to design the test to optimize the composite composition. The optimum ratio of 3 factors, e.g. chemically modified corn straw (0.10 to 0.40 g), chitosan (0.25 to 0.75 g), and poly(lactic acid) (2.00 to 3.00 g) on the response value (bending strength) of the composite was investigated. RSM-BBD provided the optimum combination of composites. The novel composite prepared under the optimized factors was characterized by Fourier transform infrared spectroscopy, mechanical testing, water absorption tests, contact angle tests, and scanning electron microscopy. The results showed that the mechanical strength, e.g., bending strength, impact strength, and tensile strength of chemically modified corn straw based composite were 21.6 MPa, 4.43 kJ/m2, and 20.0 MPa, respectively, which increased by 23.5%, 13.9%, and 18.7% compared to native corn straw based composite. Improved mechanical strength and hydrophobicity for chemically modified corn straw/chitosan/poly(lactic acid) demonstrated that chemically modified biomass fibers and bio-based degradable polymers have the potential to produce environmentally friendly composites.

Abstract Oxygen evolution reaction (OER) is a critical anodic reaction of electrochemical water splitting, developing a high‐efficiency electrocatalyst is essential. Transition metal‐based catalysts are much more cost‐effective if comparable activities can be achieved. Among them, fluorides are rarely reported due to their low aqueous stability of coordination and low electric conductivity. Herein, a NiCo bimetallic fluoride with good crystallinity is designed and constructed, and significantly enhanced catalytic activity and conductivity are observed. The inevitable oxidation of transition metal ions at high potential and the dissociation of F− are attributed to the low aqueous stability of coordination. The theoretical researches predicte that transition metal fluorides should have a strong tendency to electrochemical reconstruction. Therefore, based on the observations on their electrochemical behavior, high‐resolution transmission electron microscopy, X‐ray photoelectron spectroscopy, and bode plots, it is further demonstrated that surface reconstruction occurred during the electrochemical process, meanwhile a significant increase of electrochemically active area, which is created by F migration, are also directly observed. Additionally, DFT calculation results show that the electronic structure of the catalysts is modulated by the bimetallic centers, and this reconstruction helps optimizing the adsorption energy of oxygen‐containing species and improves OER activity.

Background Extensive attention has been given to the role of myeloid-derived suppressor cells (MDSCs) in driving tumor progression and treatment failure. Preclinical studies have identified multiple agents that eliminate MDSCs. However, none have been authorized in the cliniccal ues due to the safety reasons. In the present study, we investigated the efficacy and mechanism of sulforaphane (SFN) to eliminate MDSCs in the tumor microenvironment (TME).Methods We monitored SFN effect on tumor growth and the percents or apoptosis of immune cell subsets in mice models bearing LLC or B16 cells. Flow cytometry, quantitative reverse transcription-PCR, immunohistochemistry, ELISA, immunofluorescence, imaging flow cytometry and western blot were performed to validate the role of SFN on MDSCs function in vivo and in vitro. RNA sequencing was then used to interrogate the mechanisms of how SFN regulated MDSCs function. Tumor xenograft models were established to evaluate the involvement of IL-12RB2/MMP3/FasL induced MDSCs apoptosis in vivo. We verified the effect of SFN on MDSCs and CD8+ T cells in the blood samples from a phase I clinical trial (KY-2021–0350).Results In this study, we elucidated that SFN liberated CD8+ T-cell antitumor ability by reducing MDSCs abundance, leading to repressed tumor growth. SFN treatment suppressed MDSCs accumulation in the peripheral blood and tumor sites of mice, but had no effect on the bone marrow. Mechanistically, SFN activates IL-12RB2, which stimulates the MMP3/FasL signaling cascade to trigger caspase 3 cleavage and induce apoptosis in MDSCs. Clinically, SFN treatment eliminates peripheral MDSCs and increases the percentage and activation of CD8+ T cells.Conclusions Collectively, we uncovered the role of SFN in eliminating MDSCs to emancipate CD8+ T cells through IL-12RB2/MMP3/FasL induced apoptosis, thus providing a strategy for targeting MDSCs to control tumors and improve clinical efficacy.

A new class of bio-composites was developed by utilizing four kinds of lignocellulosic biomass fiber (bagasse, bamboo, rice husk, and rice straw) as filling fibers. Poly-β-hydroxybutyrate (PHB) and poly(butylene succinate) (PBS) in a mixture ratio of 7:3 were used as matrix materials with hot-press molding. The performance of the resulting composites was evaluated by compositional analyses, mechanical analysis, Fourier transform infrared (FTIR) spectroscopy, thermogravimetry, and morphological analysis. The interfacial adhesion, thermal stability, and comprehensive mechanical properties of the alkali treated bamboo/PHB/PBS composite were highest among the four bio-composites. The bending strength, tensile strength, and impact strength for alkali treated bamboo/PHB/PBS composite was 19.82 MPa, 12.97 MPa, and 4.30 kJ/m2, respectively. The thermal stability for NaOH modified bamboo/PHB/PBS composite was slightly superior to the other three composites, with the initial pyrolysis temperature of 248 °C, moderate pyrolysis speed, and the amount of pyrolysis residue (5.81%). The results showed the suitability of biomass fiber and biodegradable polymer for producing environmentally friendly composite materials.

Alkali-treated bagasse fiber was used as a process variable for optimization of the properties of polybutylene succinate/poly(lactic acid)-based biocomposites using Box-Behnken design (BBD) and response surface methodology (RSM). The optimum conditions for three factors, i.e., NaOH-treated bagasse fiber (0.55 to 1.65 g), polybutylene succinate (1.1 to 2.3 g), and poly(lactic acid) (2.2 to 3.4 g) on the bending strength of biocomposite were investigated. The optimum combination was 0.91 g of NaOH-treated bagasse fiber, 1.14 g of polybutylene succinate, and 3.10 g of poly(lactic acid). The bending strength for NaOH-treated bagasse fiber/polybutylene succinate/ poly(lactic acid) composite was 27.0 MPa, which was 26.0% higher than native bagasse fiber-based composite. The composites were also characterized by thermogravimetric analysis, mechanical testing, Fourier transform infrared, scanning electron microscopy, water absorption, and contact angle tests. Results demonstrated that the bending strength, impact strength, and tensile strength of alkali treated bagasse fiber-based biocomposite increased by 26.0%, 15.5%, and 23.3%, separately, compared with native bagasse-based composite after sequential homogenization, compounding, and hot pressing. The hydrophobicity for alkali-treated bagasse fiber/PBS/PLA was also improved. Thus, NaOH-treated biomass materials/biodegradable polymer was judged to be suitable for preparing green composite materials.

Abstract Myeloid-derived suppressor cells (MDSCs) were found to gradually accumulate in the orthotopic esophageal cancer mouse model during tumor progression. Although the roles of MDSCs in promoting tumor growth and inhibiting immune response have been extensively explored, currently, there are still no effective means for targeting MDSCs clinically. The deficiency of specific markers of MDSCs was responsible for the limited strategy to eliminating in clinic. This study identified that GPR84 was exclusively overexpressed on MDSCs. It was further found that GPR84 was prominently expressed on MDSCs in clinical samples and tumor mouse models, which drives the immunosuppression on CD8+T cells by inhibiting PD-L1 degradation in lysosomes. Furthermore, G-CSF and GM-CSF were found to induce GPR84 expression through the STAT3/C/EBPβ signaling pathway. In addition, GPR84+MDSCs and PD-L1+MDSCs were highly accumulated in anti-PD-1 therapy-resistant patients with esophageal cancer, and high GPR84 signature risk was verified as a negative factor for the overall survival of patients with anti-PD-1 treatment. Finally, GPR84 antagonism combined with an anti-PD-1 antibody enhanced the antitumor responses. Therefore, targeting GPR84 enhanced anti-PD-1 efficacy in esophageal cancer and other malignant tumors. This combination therapy has the potential for tumor therapy in clinics.

Backgrounds G-protein-coupled receptor 84 (GPR84) marks a subset of myeloid-derived suppressor cells (MDSCs) with stronger immunosuppression in the tumor microenvironment. Yet, how GPR84 endowed the stronger inhibition of MDSCs to CD8+ T cells function is not well established. In this study, we aimed to identify the underlying mechanism behind the immunosuppression of CD8+ T cells by GPR84+ MDSCs.Methods The role and underlying mechanism that MDSCs or exosomes (Exo) regulates the function of CD8+ T cells were investigated using immunofluorescence, fluorescence activating cell sorter (FACS), quantitative real-time PCR, western blot, ELISA, Confocal, RNA-sequencing (RNA-seq), etc. In vivo efficacy and mechanistic studies were conducted with wild type, GPR84 and p53 knockout C57/BL6 mice.Results Here, we showed that the transfer of GPR84 from MDSCs to CD8+ T cells via the Exo attenuated the antitumor response. This inhibitory effect was also observed in GPR84-overexpressed CD8+ T cells, whereas depleting GPR84 elevated CD8+ T cells proliferation and function in vitro and in vivo. RNA-seq analysis of CD8+ T cells demonstrated the activation of the p53 signaling pathway in CD8+ T cells treated with GPR84+ MDSCs culture medium. While knockout p53 did not induce senescence in CD8+ T cells treated with GPR84+ MDSCs. The per cent of GPR84+ CD8+ T cells work as a negative indicator for patients’ prognosis and response to chemotherapy.Conclusions These data demonstrated that the transfer of GPR84 from MDSCs to CD8+ T cells induces T-cell senescence via the p53 signaling pathway, which could explain the strong immunosuppression of GPR84 endowed to MDSCs.

ObjectiveThe aim of this study was to explore the safety and efficacy of multiple peptide-pulsed autologous dendritic cells (DCs) combined with cytotoxic T lymphocytes (CTLs) in patients with cancer.MethodsFive patients diagnosed with cancer between November 2020 and June 2021 were enrolled and received DC-CTLs therapy. Peripheral blood was collected and antigenic peptides were analyzed. The phenotype and function of DC-CTLs and the immune status of patients were detected using flow cytometry or IFN-γ ELISPOT analysis.ResultsDCs acquired a mature phenotype and expressed high levels of CD80, CD86, CD83, and HLA-DR after co-culture with peptides, and the DC-CTLs also exhibited high levels of IFN-γ. Peripheral blood mononuclear cells from post-treatment patients showed a stronger immune response to peptides than those prior to treatment. Importantly, four of five patients maintained a favorable immune status, of which one patient’s disease-free survival lasted up to 28.2 months. No severe treatment-related adverse events were observed.ConclusionOur results show that multiple peptide-pulsed DCs combined with CTLs therapy has manageable safety and promising efficacy for cancer patients, which might provide a precise immunotherapeutic strategy for cancer.

Increasing pollution has significantly threatened the Antarctic ecosystem. The contamination of hydrocarbons has drawn a considerable amount of attention owing to their toxicity, recalcitrance, and persistence. Considering the Antarctic Treaty, only indigenous species are allowed to bioremediate the contaminated environment. However, the knowledge of the ecological role, physiology, function, and genomics of endemic hydrocarbon consumers is still limited. Here, we investigated the dynamics of phenanthrene-consuming communities derived from the Antarctic soil and found that Variovorax, Rhodocyclaceae, and Hydrogenophaga were differentiated in all the phenanthrene-consuming subcultures. We isolated a pure culture of the key hydrocarbon consumer Variovorax sp. strain N23. Moreover, the result of the polyphasic approach suggested that strain N23 represents a novel species of the genus Variovorax. In addition, the genomic characteristics of this strain revealed incomplete degradation pathways for diverse hydrocarbons. Overall, this study reveals the relatively weak hydrocarbon-degrading potential of the indigenous bacteria and suggests the need for more careful protection of the Antarctic ecosystem.

Marine fungi, such as species from the Penicillium and Aspergillus genera, are prolific producers of a diversity of natural products with cytotoxic properties. These fungi have been successfully isolated and identified from various marine sources, including sponges, coral, algae, mangroves, sediment, and seawater. The cytotoxic compounds derived from marine fungi can be categorized into five distinct classes: polyketides, peptides, terpenoids and sterols, hybrids, and other miscellaneous compounds. Notably, the pre-eminent group among these compounds comprises polyketides, accounting for 307 out of 642 identified compounds. Particularly, within this collection, 23 out of the 642 compounds exhibit remarkable cytotoxic potency, with IC50 values measured at the nanomolar (nM) or nanogram per milliliter (ng/mL) levels. This review elucidates the originating fungal strains, the sources of isolation, chemical structures, and the noteworthy antitumor activity of the 642 novel natural products isolated from marine fungi. The scope of this review encompasses the period from 1991 to 2023.

The high cost and carbon footprint of conventional alkali activator (sodium silicate) limit the wide-scale adaptation of alkali-activated materials. During the production of sodium silicate(SS), carbonate and silicate need to be calcined at 1400 °C, which is not environmentally friendly. In order to improve the sustainability of the activator, 10% SS was reduced in this paper. The effect alternative silicon sources (ASS) on the mechanical properties of alkali-activated carbon steel slag (CSS) prepared at room temperature were studied when the modulus is 0.5 and 1.0, and the mechanism of silicon source substitution was discussed by means of microscopic characterization. The substitution amount of ASS was 10% of the total CSS amount. ASS included waste glass powder (WGP), rice husk ash (RHA), diatomaceous earth (DE), diatomaceous earth analytical reagent (DEAR), and silica fume (SF). Microscopic characterization methods included X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), thermogravimetry and differential thermogravimetry (TG-DTG), and scanning electron microscope-energy dispersive spectrometer (SEM-EDS). The results show that the effect of ASS depended on the structure of SiQn, and is affected by the initial modulus of the activator. The compressive strengths of the low modulus samples are lower than that of the high modulus samples, while the effect of silicon source substitution is the opposite. However, the effect of SF is less affected by the initial modulus, and the 90d compressive strength of low modulus and high modulus differed by only 8%. 10% SF substitution can save at least 48.28% of industrial SS. SF is an economic and effective ASS, which can provide a better ecological choice for the production of alkali-activated CSS cementitious materials.

Background: Readmission shortly after discharge is indicative of an increased disease severity for patients with ulcerative colitis (UC) and ineffectiveness to medical therapy, which may contribute to a dismal prognosis. Objectives: This study aimed to explore prognostic variables with a nomogram to predict unplanned UC-related readmission within 1 year after discharge. Design: A retrospective cohort study. Methods: Electronic medical records of all UC patients treated at our center between 1 January 2014 and 31 June 2021 were reviewed. A comprehensive analysis of various characteristics, such as demographics, comorbidities, medical history, follow-up appointments, admission endoscopy, histopathologic features, etc., was used to determine the primary end point, which was unplanned UC-related calendar year readmission. Results: We found that the unplanned UC-related readmission rate within 1 year was 20.8%. In multivariable cox analysis, the predictors of the Elixhauser comorbidity index [Hazard ratio (HR): 3.50, 95% confidence interval (CI): 1.93–6.37], regular follow-up (HR: 0.29, 95% CI: 0.16–0.53), any history of corticosteroid use (HR: 3.38, 95% CI: 1.83–6.27), seral level of C-reactive protein (HR: 1.01, 95% CI: 1.00–1.02), and the UC endoscopic index of severity (HR: 1.29, 95% CI: 1.05–1.57) independently predicted calendar year readmission after discharge. The established nomogram had a consistently high accuracy in predicting calendar year readmission in the training cohort, with a concordance index of 0.784, 0.825, and 0.837 at 13, 26, and 52 weeks, respectively, which was validated in both the internal and external validation cohorts. Therefore, UC patients were divided into clinically low-, high-, and extremely high-risk groups for readmission, based on the calculated score of 272.5 and 378. Conclusion: The established nomogram showed good discrimination and calibration powers in predicting calendar year readmission in high-risk UC patients, who may need intensive treatment and regular outpatient visits.

Hydrocarbons are ubiquitous in marine environments and might fuel hydrocarbon-metabolizing microbes in the ocean. Numerous studies have documented microbial hydrocarbon degradation in water columns and deep-sea surface sediment. However, the degradation potential and biogeochemical cycling of hydrocarbons in subsurface sediments remain largely unknown. In this study, we used two different hydrocarbons, n-hexadecane (HEX) and methylcyclohexane (MCH), to investigate the distribution and diversity of hydrocarbon-consuming bacteria in a core sediment sample from the Central Indian Ridge (CIR), which is adjacent to mid-ridge hydrothermal vents in the Indian Ocean. We observed different vertical profiles of HEX- and MCH-degrading bacteria in the core sediments. Specifically, HEX-degrading bacteria were universally distributed, while MCH-degrading bacteria were found only in the intermediate layers of the core sediments. Changing factors including dissolved oxygen might affect the natural distribution of different hydrocarbon consumers. We found that a novel species of the genus C1-B045 might play a pivotal role in metabolizing MCH in the CIR deep biosphere. Through amino acid identity comparison with published sequences, we determined that C1-B045 harbors two novel classes of cyclohexanone monooxygenases involved in MCH metabolism. This study sheds light on the structure and function of hydrocarbon-consuming microbes in deep biospheres.

Antiretroviral therapy alone is insufficient in curing HIV-1 infection, due to latent viral reservoir persistency. Here, authors explore the post-virologic control of combining active and passive immunisation with vesatolimod, in a SHIV-infected rhesus macaque model.

Abstract Cancer is a complicated disease attributed to multifactorial changes, which causes difficulties with treatment strategies. Various factors have been regarded as the main contributors, and infectious etiological factors have recently attracted interest. Several microbiomes contribute to carcinogenesis, cancer progression, and modulating cancer treatment by inducing cancerous epithelial cells and chronic inflammation. Most of our knowledge on the role of microbiota in tumor oncogenesis and clinical efficiency is associated with the intestinal microbiome. However, compelling evidence has also confirmed the contribution of the intratumor microbiome in cancer. Indeed, the findings of clinical tumor samples, animal models, and studies in vitro have revealed that many intratumor microbiomes promote tumorigenesis and immune evasion. In addition, the intratumor microbiome participates in regulating the immune response and even affects the outcomes of cancer treatment. This review summarizes the interplay between the intratumor microbiota and cancer, focusing on the contribution and mechanism of intratumor microbiota in cancer initiation, progression, and potential applications to cancer therapy.

Abstract Developing an intervention that results in virologic control following discontinuation of antiretroviral therapy (ART) is a major objective of HIV-1 cure research. In this study, we investigated the therapeutic efficacy of a vaccine consisting of adenovirus serotype 26 (Ad26) and modified vaccinia Ankara (MVA) with or without an SIV Envelope (Env) gp140 protein with alum adjuvant in combination with the TLR7 agonist vesatolimod (GS-9620) in 36 ART-suppressed, SIVmac251-infected rhesus macaques. Ad26/MVA therapeutic vaccination led to robust humoral and cellular immune responses, and the Env protein boost increased antibody responses. Following discontinuation of ART, virologic control was observed in 5/12 animals in each vaccine group, compared with 0/12 animals in the sham control group. These data demonstrate therapeutic efficacy of Ad26/MVA vaccination with vesatolimod but no clear additional benefit of adding an Env protein boost. SIV-specific cellular immune responses correlated with virologic control. Our findings show partial efficacy of therapeutic vaccination following ART discontinuation in SIV-infected rhesus macaques.

Meningitis-like infectious disease (MID) (also known as frog cataract and torticollis) is a disease prone to occur in amphibians and reptiles. It is highly contagious and has a high mortality rate. In this study, we sampled and sequenced microbiomes from oral and intestinal samples of five normal and five diseased bullfrogs. The analysis found that the richness, uniformity, and abundance of the microbial community of the diseased bullfrogs were significantly higher than those of the normal bullfrogs in both the oral cavity and the gut. In the diseased group, the abundance of Elizabethkingia significantly increased and that of Lactococcus significantly decreased. It showed that the structure of the microbial community had changed a lot in diseased frogs. After the pathogenic bacteria infected the body, it might be make the decline in the immune function of the body declined, and resulting in some conditional pathogenic bacteria in the water body further infecting the body. As a result, the richness and composition of the microbial community significantly changed. This study can provide a theoretical basis for the control of MID of bullfrogs.

Abstract Background Osteosarcoma (OS) is the most common malignant tumor of bone, and the clinical efficacy of current treatments and associated survival rates need to be further improved by employing novel therapeutic strategies. Although various studies have shown that BMI1 protein is universally upregulated in OS cells and tissues, its specific role and underlying mechanism have not yet been fully explored. Methods Expression of BMI1 protein in OS cells was detected by western blot. The effect of BMI1 on proliferation and migration of OS cells (143B and U-2OS cell lines) was investigated in vitro using CCK-8, colony formation and transwell assays, and in vivo using subcutaneous tumorigenesis and lung metastasis assays in xenograft nude mice. Expression of epithelial–mesenchymal transition (EMT)-associated proteins was detected by immunofluorescence imaging. Bioinformatic analysis was performed using ENCODE databases to predict downstream targets of BMI1. SIK1 mRNA expression in osteosarcoma cells was detected by quantitative real-time reverse transcription PCR (qPCR). Chromatin immunoprecipitation-qPCR (ChIP-qPCR) was used to investigate expression of BMI1-associated, RING1B-associated, H2AK119ub-associated and H3K4me3-associated DNA at the putative binding region of BMI1 on the SIK1 promoter in OS cells. Results Using both in vitro and in vivo experimental approaches, we found that BMI1 promotes OS cell proliferation and metastasis. The tumor suppressor SIK1 was identified as the direct target gene of BMI1 in OS cells. In vitro experiments demonstrated that SIK1 could inhibit proliferation and migration of OS cells. Inhibition of SIK1 largely rescued the altered phenotypes of BMI1-deficient OS cells. Mechanistically, we demonstrated that BMI1 directly binds to the promoter region of SIK1 in a complex with RING1B to promote monoubiquitination of histone H2A at lysine 119 (H2AK119ub) and inhibit H3K4 trimethylation (H3K4me3), resulting in inhibition of SIK1 transcription. We therefore suggest that BMI1 promotes OS cell proliferation and metastasis by inhibiting SIK1. Conclusions Our results reveal a novel molecular mechanism of OS development promoted by BMI1 and provides a new potential target for OS treatment.

Inosine 5′-monophoaphate (IMP) is a food additive that promotes serious lipohyperplasia in the liver of C57/KsJ-db/db (db/db) mice. Thus, IMP taken orally by healthy mice might also damage their health. To date, how IMP affects health after being taken by healthy animals is still unclear. Therefore, we investigated the health of C57BL/6J mice affected by IMP intake. Our data revealed that C57BL/6J mice administered 255 μM IMP daily via oral gavage for 4 months caused hyperlipidemia and an increase in body fat rate. The expressions of acetyl-CoA carboxylase 1 (ACC1) and phosphorylated acetyl-CoA carboxylase 2 (ACC2) in hepatocytes increased though the administration of IMP, promoting the phosphorylation of adenosine 5′-monophosphate-activated protein kinase (AMPK). The conversion of acetyl-CoA into triglycerides (TGs) was promoted by ACC1. These TGs were transported from the hepatocytes to avoid the development of non-alcoholic fatty liver disease (NAFLD), causing a deficiency of acetyl-CoA in the liver, and then, the increased phosphorylated ACC2 promoted the cytoplasm fatty acids entering the mitochondria and conversion into acetyl-CoA through the fatty acid β-oxidation pathway, causing a deficiency in fatty acids. Therefore, the liver showed enhanced absorption of exogenous fatty acids, which were converted into TGs, causing lipohyperplasia. In conclusion, an excessive IMP intake promotes metabolic dysfunction in adipose tissue.

Abstract Microporous structure plays a significant role in bone tissue engineering, to influence inductive bone formation and elevate bone ingrowth inside microporous scaffolds. We hereby fabricated microporous scaffolds by 3D printing with mixture of porogens and photopolymerizing resin. Our method can tune the microporosity of scaffolds from 0% to 36% by varying the porogen concentration in the inks from zero to 60 vol%. The microporosity and microspore size of scaffolds can affect in vitro expansion of preosteoblast cells. We evaluated the attachment, spreading and proliferation of MC3T3‐E1 mouse preosteoblast cells on the printed porous scaffolds. Our studies revealed that preosteoblast cells’ in vitro adhesion and proliferation were significantly mediated by the printed scaffolds. Cells proliferation on scaffolds with 20%–30% microporosity showed much higher rate than on other scaffolds. In this microporosity range, scaffolds also showed much better cell spreading and morphologies. At a low level of microporosity ( 35%) led to very poor cell attachment and is unfavorable for the proliferation of MC3T3‐E1 cells. Furthermore, the printed dual macro‐/micro‐porous scaffolds showed higher proliferation rate of MC3T3‐E1 cells as compared to mono‐pore sized scaffolds, either the macro‐porous or microporous structure.

Abstract The a priori T cell repertoire and immune response against SARS-CoV-2 viral antigens may explain the varying clinical course and prognosis of patients having a mild COVID-19 infection as opposed to those developing more fulminant multisystem organ failure and associated mortality. Using a novel SARS-Cov-2-specific artificial antigen presenting cell (aAPC), coupled with a rapid expansion protocol (REP) as practiced in tumor infiltrating lymphocytes (TIL) therapy, we generate an immune catalytic quantity of Virus Induced Lymphocytes (VIL). Using T cell receptor (TCR)-specific aAPCs carrying co-stimulatory molecules and major histocompatibility complex (MHC) class-I immunodominant SARS-CoV-2 peptide-pentamer complexes, we expand virus-specific VIL derived from peripheral blood mononuclear cells (PBMC) of convalescent COVID-19 patients up to 1000-fold. This is achieved in a clinically relevant 7-day vein-to-vein time-course as a potential adoptive cell therapy (ACT) for COVID-19. We also evaluate this approach for other viral pathogens using Cytomegalovirus (CMV)-specific VIL from donors as a control. Rapidly expanded VIL are enriched in virus antigen-specificity and show an activated, polyfunctional cytokine profile and T effector memory phenotype which may contribute to a robust immune response. Virus-specific T cells can also be delivered allogeneically via MHC-typing and patient human leukocyte antigen (HLA)-matching to provide pragmatic treatment in a large-scale therapeutic setting. These data suggest that VIL may represent a novel therapeutic option that warrants further clinical investigation in the armamentarium against COVID-19 and other possible future pandemics.

PurposeGross motor skills (GMS) and inhibitory control (IC) which are both development in preschool stage is significant for preschooler to healthy growth. However, the evidence of relationship between them in preschoolers are still insufficient, most of studies only focus on youth. Thus, the aim of this research is to examine the association between GMS and IC in preschool children.MethodsThis cross-sectional study used baseline data from a previous intervention study of preschoolers conducted in 2018. GMS were assessed by using the Test for Gross Motor Development (2nd edition) in preschoolers, which includes two subtests of locomotor and object control skills. Total GMS is calculated from the sum of these two subtests. The Fish Flanker task was used to evaluate both accuracy and reaction time of IC. Multivariate linear regression models were established to analyze the relationships between GMS and IC.ResultsA total of 123 preschool-age children (55 girls, 68 boys) were included in the final analysis. After adjusting for confounders, GMS (β = −8.27 ms, 95%CI: −14.2, −2.34), locomotor (β = −11.2 ms, 95%CI: −21.43, −0.97), and object control skills (β = −12.15 ms, 95%CI: −22.07, −2.23) were all negatively related with reaction time of IC.ConclusionThere was a significant negative correlation between gross motor skills and the reaction time of inhibitory control in preschool children. Further research is needed to verify this finding in prospective and experimental studies.

Abstract Background Tumor necrosis factor-associated apoptosis-inducing ligand (TRAIL) was initially considered an immunity guard; however, its function remains controversial. Besides immune cells, lung and colon cancer cells have also been reported to express TRAIL, which can promote tumor invasion and metastasis. However, the biological function and underlying mechanism of action of TRAIL in esophageal squamous cell carcinoma (ESCC) remain poorly elucidated. Methods The ESCC cells stemness, migration, and proliferation ability was assessed by sphere formation, Transwell, and CCK8 assay. The stemness- and epithelial-mesenchymal transition (EMT)- related genes expression levels were analyzed by Western blot and RT-qPCR. The signal activation was conducted by Western blot. The xenograft mouse experiments and lung metastasis model were performed to confirm our findings in vitro. Results Herein, we found that TRAIL is a negative predictor in patients with ESCC. To further investigate the biological function of TRAIL, we established TRAIL knockdown and overexpression ESCC cell lines and found that TRAIL induced EMT and promoted tumor aggressiveness. Furthermore, we demonstrated that TRAIL- overexpressing cells upregulated PD-L1 expression, which was dependent on the p-ERK/STAT3 signaling pathway. We obtained similar results when using recombinant human TRAIL. Finally, we validated the biological role and mechanism of action of TRAIL in vivo. Conclusions These findings demonstrate that TRAIL promotes ESCC progression by enhancing PD-L1 expression, which induces EMT. This may explain the failure of TRAIL preclinical trials.

BackgroundTo provide a reference for hospital drug selection and rational clinical drug selection based on the evaluation of the safety, nutritional quality, and economy of 27 manufacturers of five varieties (18AA, 18AA-I, 18AA-II, 18AA-IV, 18AA-V) of balanced compound amino acids for injection and (18AA-IIoriginal research).MethodsThe safety of compound amino acids for injection was evaluated by comparing the antioxidant sulfite contents. Based on the amino acid scoring standard mode and the whole egg protein mode as proposed by the Food and Agriculture Organization of the United Nations/World Health Organization (FAO/WHO) in 1973, we compared the formula. The first limiting amino acid content and the comprehensive quality of the total essential amino acid (EAA) contents of the six formulations were studied. The price/content ratio was used to evaluate their economy.ResultsSimilar variety produced by different manufacturers have the same formula and contents of balanced compound amino acids for injection. Safety: 18AA-IIoriginal research and 18AA-II had the lowest sulfite content. Compared with 18AA-IIoriginal research, the sulfite content of 18AA-I, 18AA, 18AA-V, and 18AA-IV were higher (10 times, 16.67 times, 16.67 times, and 33.33 times, respectively). The lower the sulfite content, the safer the product. Nutritional quality: The proportions of amino acids in the five varieties of compound amino acid injection were all suitable. The order of the first limiting amino acids for the formulations was 18AA-IIoriginal research = 18AA-II>18AA >18AA-I = 18AA-IV>18AA-V. The order of the EAA values for the formulations was 18AA-IIoriginal research = 18AA-II>18AA>18AA-I > 18AA-IV > 18AA-V. The overall effectiveness order was 18AA-IIoriginal research = 18AA-II>18AA > 18AA-I>18AA-IV>18AA-V. Economy: Among the 27 manufacturers, 12 manufacturers had a price/content ratio higher than that of 18AA-II original research manufacturers, and 15 manufacturers had a price/content ratio lower than original research manufacturers.ConclusionThrough its security, effectiveness, and economy of the comprehensive research, we recommended 18AA-II and 18AA-IIoriginal research with high safety, efficacy, and reasonable price as the first choice. 18AA and 18AA-I with better safety and reasonable price, secondary recommendation. 18AA-IV or 18AA-V with poor safety, efficacy, and economy are not recommended.

Despite the rapid creation of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) vaccines, the precise correlates of immunity against severe Coronavirus Disease 2019 (COVID-19) are still unknown. Neutralizing antibodies represent a robust surrogate of protection in early Phase III studies, but vaccines provide protection prior to the evolution of neutralization, vaccines provide protection against variants that evade neutralization, and vaccines continue to provide protection against disease severity in the setting of waning neutralizing titers. Thus, in this study, using an Ad26.CoV2.S dose-down approach in nonhuman primates (NHPs), the role of neutralization, Fc effector function, and T-cell immunity were collectively probed against infection as well as against viral control. While dosing-down minimally impacted neutralizing and binding antibody titers, Fc receptor binding and functional antibody levels were induced in a highly dose-dependent manner. Neutralizing antibody and Fc receptor binding titers, but minimally T cells, were linked to the prevention of transmission. Conversely, Fc receptor binding/function and T cells were linked to antiviral control, with a minimal role for neutralization. These data point to dichotomous roles of neutralization and T-cell function in protection against transmission and disease severity and a continuous role for Fc effector function as a correlate of immunity key to halting and controlling SARS-CoV-2 and emerging variants.

The origin and nature of rebound HIV-1 virus following antiretroviral therapy (ART) discontinuation still remains unclear. Here, Liu et al. suggest that intact proviral DNA in peripheral blood and lymph node mononuclear cells during ART suppression likely is the source of viral rebound following ART discontinuation.

Rationale: Nonalcoholic fatty liver disease (NAFLD) is a kind of metabolic disease characterized by liver steatosis. Excessive reactive oxygen species (ROS) originating from dysfunctional mitochondria is the major pathophysiological contributor in the development of NAFLD and is thought to be a promising therapeutic target. A few reports demonstrate the antioxidative treatments for NAFLD.Methods: Male C57 mice were fed on a normal chow diet (ND) or high-fat diet (HFD) for 8 weeks. PBS or N-acetyl cysteine (NAC) was gavaged to mice. LO2 human liver cell line treated with palmitic acid (PA) was applied as a cellular model. Western blot, immunofluorescence, biochemistry assay, and pathological staining were used to investigate the mechanism of suppressing lipid accumulation of NAC.Results: NAC treatment was able to prevent HFD-induced NAFLD, as evidenced by less hepatic triglyceride accumulation and lipid droplet formation compared with that of mice in the HFD group. NAC could preserve mitochondrial function by inhibiting excessive mitophagy and promoting mitochondria biogenesis to prevent ROS production. NAC also activated Sirt1 and preserved its protein level and subsequently promoted mitochondria biogenesis via deacetylating PGC1a.Conclusion: We demonstrated that NAC may be an effective drug to treat NAFLD, which was related to its antioxidative and mitochondrial protective effect.

Obesity, a global public health issue, is characterized by excessive adiposity and is strongly related to some chronic diseases including cardiovascular diseases and diabetes. Extra energy intake-induced adipogenesis involves various transcription factors and long noncoding RNAs (lncRNAs) that control lipogenic mRNA expression. Currently, lncRNAs draw much attention for their contribution to adipogenesis and adipose tissue function. Increasing evidence also manifests the pivotal role of lncRNAs in modulating white, brown, and beige adipose tissue development and affecting the progression of the diseases induced by adipose dysfunction. The aim of this review is to summarize the roles of lncRNAs in adipose tissue development and obesity-caused diseases to provide novel drug targets for the treatment of obesity and metabolic diseases.

Abstract Backgroud Horizontal gene transfer and gene duplication are two major mechanisms contributing to the evolutionary adaptation of organisms. Previously, polygalacturonase genes (PGs) were independently horizontally transferred and underwent multiple duplications in insects (e.g., mirid bugs and beetles). Here, we chose three phytozoophagous mirid bugs (Adelphocoris suturalis, A. fasciaticollis, A. lineolatus) and one zoophytophagous mirid bug (Nesidiocoris tenuis) to detect whether the duplication, molecular evolution, and expression levels of PGs were related to host range expansion in mirid bugs. Results By RNA-seq, we reported 30, 20, 19 and 8 PGs in A. suturalis, A. fasciaticollis, A. lineolatus and N. tenuis, respectively. Interestingly, the number of PGs was significantly positive correlation to the number of host plants (P = 0.0339) in mirid bugs. Most PGs (> 17) were highly expressed in the three phytozoophagous mirid bugs, while only one PG was relatively highly expressed in the zoophytophagous mirid bug. Natural selection analysis clearly showed that a significant relaxation of selection pressure acted on the PGs in zoophytophagous mirid bugs (K = 0.546, P = 0.0158) rather than in phytozoophagous mirid bugs (K = 1, P = 0.92), suggesting a function constraint of PGs in phytozoophagous mirid bugs. Conclusion Taken together with gene duplication, molecular evolution, and expression levels, our results suggest that PGs are more strictly required by phytozoophagous than by zoophytophagous mirid bugs and that the duplication of PGs is associated with the expansion of host plant ranges in mirid bugs.

In chimeric antigen receptor (CAR)-T cell therapy, the role and mechanism of indoleamine 2, 3 dioxygenase 1 (IDO1) in enhancing antitumor immunity require further study. IDO1 is one of the most important immunosuppressive proteins in esophageal squamous cell carcinoma (ESCC). However, the IDO1 inhibitor, epacadostat, has failed in phase III clinical trials; its limited capacity to inhibit IDO1 expression at tumor sites was regarded as a key reason for clinical failure. In this study, we innovatively loaded the IDO1 inhibitor into hyaluronic acid-modified nanomaterial graphene oxide (HA-GO) and explored its potential efficacy in combination with CAR-T cell therapy. We found that inhibition of the antitumor effect of CAR-T cells in ESCC was dependent on the IDO1 metabolite kynurenine. Kynurenine could suppress CAR-T cell cytokine secretion and cytotoxic activity. Inhibiting IDO1 activity significantly enhanced the antitumor effect of CAR-T cells in vitro and in vivo. Our findings suggested that IDO1 inhibitor-loaded nanosheets could enhance the antitumor effect of CAR-T cells compared with free IDO1 inhibitor. Nanosheet-loading therefore provides a promising approach for improving CAR-T cell therapeutic efficacy in solid tumors.

Background Tumor relapse due to mutation in CD19 can hinder the efficacy of chimeric antigen receptor (CAR)-T cell therapy. Herein, we focused on lymphoma patients whose B cells exhibited a point mutation in CD19 of B cells after CAR-T cell infusion.Methods The CAR-T and CD19+ B cells from peripheral blood or bone marrow were assessed using flow cytometry. Genome sequencing was conducted to identify the molecular characteristics of CAR-T and CD19+ B cells from pre-relapse and postrelapse samples. CD19 in CARs comprising single chain fragments variable (scFV) antibody with FMC63 or 21D4 was constructed. The cytotoxic efficacy of CAR-T cells was also evaluated via in vitro and in vivo experiments.Results A patient with high-grade B cell lymphoma exhibited complete response, but the lymphoma relapsed in her left breast at 6 months after CD19 CAR (FMC63)-T cell infusion. A mutation was found in exon 3 of CD19 (p.163. R-L) in malignant B cells of the patient. In two lymphoma patients who exhibited resistance to CAR-T cell therapy, a mutation was detected in exon 3 of CD19 (p.174. L-V). Functional analysis revealed that FMC63 CAR-T cells exhibited antitumor ability against wild-type CD19+ cells but were unable to eradicate these two types of mutated CD19+ cells. Interestingly, 21D4 CAR-T cells were potentially capable of eradicating these mutated CD19+ cells and exhibiting high antitumor capacity against CD19+ cells with loss of exon 1, 2, or 3.Conclusions These findings suggest that point mutation can facilitate immune escape from CAR-T cell therapy and that alternative CAR-T cells can effectively eradicate the mutated B cells, providing an individualized therapeutic approach for lymphoma patients showing relapse.

HHLA2, a newly identified B7 family member, regulates T cell functions. However, the expression and prognostic value of HHLA2 in solid tumors is ill defined. This study aimed to reveal the expression landscape of HHLA2 in various solid tumors, and to evaluate its prognostic value in kidney clear cell carcinoma (KIRC). Using The Cancer Genome Atlas (TCGA) database, we investigated the expression pattern of HHLA2 across 22 types of cancer. HHLA2 and CD8 protein expression was determined via immunohistochemistry (IHC). KIRC-specific findings were further analyzed with R software and the prognostic value was validated on tissue microarrays. HHLA2 was widely expressed in cancers at both the mRNA and protein levels. Among all tested tumors, KIRC showed the highest transcript level of HHLA2, and HHLA2 levels were significantly higher in tumor tissues than in matched normal samples, as evidenced by both TCGA and IHC data. HHLA2 was also positively correlated with survival rates in KIRC based on TCGA and clinical data. Receiver operating characteristic curves data showed the prognostic value of HHLA2 for patients with KIRC in TCGA. Moreover, HHLA2 was positively correlated with immune-related genes, while HHLA2 and CD8 expression exhibited a consistent trend in KIRC tumor samples. In conclusion, HHLA2 is highly expressed in KIRC and predicts a favorable survival outcome, highlighting that it may work as a potential target for KIRC therapy.

The piezoelectric materials with excellent bioactivity have attracted more attentions recently and have broad potential applications in tissue engineering. In this article, the barium titanate (BT) particles were filled into the polycaprolactone (PCL)/calcium sulfate whisker (CSW) (15 wt%) composites to prepare the PCL/CSW/BT ternary composites. Due to the reinforcement synergy between the CSWs and the BT particles, the mechanical properties of the ternary composites were increased by 50% compared with the PCL/BT binary composites. The piezoelectric coefficient of the ternary composites is still in the range of natural bone. The ternary composite can promote the adhesion and proliferation of cells. The composites in this study have potential applications in tissue engineering.

Tumour-associated macrophages (TAMs) display an M2 phenotype that promote tumour immune escape. Here the authors show that Chloroquine (CQ), a lysosome inhibitor used against malaria, inhibits tumour growth by switching TAMs into an M1 tumor-killing phenotype by repolarizing macrophages metabolism.

This study was conducted to evaluate the effects of dietary crude protein (CP) level on faecal CP and amino acid (AA) flow and digestibility, faecal and ileal digesta microbial AA composition. Eighteen Duroc × Landrace × Yorkshire barrows (60 ± 1.43 kg) were randomly divided into 3 groups with 6 barrows in each, and fed a maize–soybean meal diet at the 10% (L-CP), 13% (M-CP) and 16% (H-CP) CP levels. The results indicated the faecal total N, CP, total AA (TAA) flow increased linearly (P

The vectors of pinewood nematode of Bursaphelenchus xylophilus (Steiner & Bührer, 1934) are mainly known as xylophagous beetles. Understanding the composition and distribution of these xylophagous beetles in host pine trees infected by PWN is critical to control the spread of PWN. In this study, we investigated the community structures of the xylophagous beetles in two main host trees in Fujian and Shandong, Pinus massoniana Lamb. and Pinus thunbergia Parl., in different stages of infection. All beetles were collected by dissecting the whole pine trees and then identified by their morphological characteristics and COI genes. The results showed that the diversity of xylophagous beetles was different not only between the two host pine trees but also among the different infection stages. The diversity of P. massoniana xylophagous beetles was significantly higher than that of P. thunbergii, and there were also significant differences in the different stages of PWN infection. In total, Scolytinae was the most common (53.70%), followed by Curculionidae (18.26%), Cerambycidae (16.31%), and Cleridae (6.04%). Monochamus alternatus, the most important vector of PWN, occupied a large niche and showed different aggregation positions during the three infection stages in both host trees. This result might be related to the resistance of bark beetles to host trees and competition with other xylophagous beetles. The community diversity of xylophagous beetles was jointly affected by both the infection stages of PWN and the spatial niche of xylophagous beetles. Knowledge of the diversity and competitive relationships among xylophagous beetles might help regulate the population dynamics of these beetles.

In order to compare the properties of alkali-activated carbon steel slag (CSS) and stainless steel slag (SSS), the effects of sodium hydroxide/sodium silicate solution mass ratio (NH/NS), liquid/solid ratio and blast furnace slag (BFS) dosage on the compressive strength, hydration products and hydration degree of CSS and SSS were studied. Furthermore, a combination of X-ray diffraction (XRD), thermo-gravimetric analysis coupled with differential thermal analysis (TGA-DTA), Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscope-energy dispersive spectrometer (SEM-EDS) were used to characterize the morphology and structure of alkali-activated CSS-BFS and SSS-BFS cementitious materials. As the results revealed, the primary hydrate of alkali-activated CSS and SSS is C-(A)-S-H with Q2 [SiO4] units, which has a low Ca/Si ratio and includes inert phases like a CaO-FeO-MnO-MgO solid solution (RO) in CSS while cuspidine, magnesiochromite etc. in SSS. More active C3S and β-C2S promote the alkali activation of CSS, whereas the less active γ-C2S hinders the depolymerization of SSS. The incorporation of BFS does not change the hydrate, whose seed effect is helpful for accelerating the depolymerization and polycondensation of CSS and SSS, especially for SSS, and makes the hydrate increase significantly. Owing to the high SiO2 and Al2O3 contents of SSS, the C-(A)-S-H chain length is increased, thus facilitating the polycondensation effect. In this study, the optimal NH/NS of CSS and SSS is NH/NS= 1:2, and the optimal liquid/solid ratio is 0.29. Compared to CSS−BFS, the C-(A)-S-H gel produced by SSS−BFS has lower Ca/Si and Al/Si ratios. Unlike CSS, pure SSS is inappropriate as an alkali-activated precursor and needs to be co-activated with BFS.

Cylindrocladium leaf blight is one of the most important diseases in Eucalyptus plantations. We investigated the proteome and transcriptome of Eucalyptus infected or not infected with Calonectria pseudoreteaudii. Here we provide the information about the processing of raw data obtained by RNA-seq and iTRAQ technologies. The data are related to [1].

A compound containing an imidazole moiety and a 2,4-dienone motif with significant activity toward several fungi was discovered in a screen for new antifungal compounds. Then, a total of 26 derivatives of this compound were designed, synthesized and evaluated through in vitro and in vivo antifungal activity assays. Several compounds exhibited improved antifungal activities compared to the lead compound. Of the derivatives, compounds 31 and 42 exhibited strong, broad-spectrum inhibitory effects toward Candida spp. In particular, the two derivatives exhibited potent antifungal activities toward the fluconazole-resistant isolate C. albicans 64110, with both having MIC values of 8 µg/mL. In addition, they had significant inhibitory effects toward two Gram-positive bacteria, Staphylococcus aureus UA1758 (compound 31: MIC = 8 µg/mL; compound 42: MIC = 4 µg/mL) and Staphylococcus epidermidis UF843 (compound 31: MIC = 8 µg/mL; compound 42: MIC = 8 µg/mL). The results of an animal experiment indicated that both compounds could improve the survival rate of model mice infected with ATCC 90028 (fluconazole-susceptible isolate). More importantly, the two compounds exhibited notable in vivo effects toward the fluconazole-resistant C. albicans isolate, which is promising with regard to the clinical problem posed by fluconazole-resistant Candida species.

In the originally published version of this Article, images in Fig. 5n were inadvertently replaced with duplicates of images in Fig. 5o during the production process. This has now been corrected in both the PDF and HTML versions of the Article.

Spider venom comprises a mixture of compounds with diverse biological activities, which are used to capture prey and defend against predators. The peptide components bind a broad range of cellular targets with high affinity and selectivity, and appear to have remarkable structural diversity. Although spider venoms have been intensively investigated over the past few decades, venomic strategies to date have generally focused on high-abundance peptides. In addition, the lack of complete spider genomes or representative cDNA libraries has presented significant limitations for researchers interested in molecular diversity and understanding the genetic mechanisms of toxin evolution. In the present study, second-generation sequencing technologies, combined with proteomic analysis, were applied to determine the diverse peptide toxins in venom of the Chinese bird spider Ornithoctonus huwena. In total, 626 toxin precursor sequences were retrieved from transcriptomic data. All toxin precursors clustered into 16 gene superfamilies, which included six novel superfamilies and six novel cysteine patterns. A surprisingly high number of hypermutations and fragment insertions/deletions were detected, which accounted for the majority of toxin gene sequences with low-level expression. These mutations contribute to the formation of diverse cysteine patterns and highly variable isoforms. Furthermore, intraspecific venom variability, in combination with variable transcripts and peptide processing, contributes to the hypervariability of toxins in venoms, and associated rapid and adaptive evolution of toxins for prey capture and defense.