Search

Stigmaria asiatica Jongmans et Gothan is a common species from the Permo-Carboniferous of East Asia. It is characterized by a relatively slender rhizomorph and represents the underground rooting system of lowland arborescent lycopsids. This species from the Wuda Coalfield (Asselian, lower Permian) represents the recovery taxon dominating the subsequent peat-forming vegetation after a volcanic eruption destroyed the previous flora, termed the Wuda Tuff Flora (Chinese “vegetational Pompeii”). It is characterized by novel downward axes, probably for access to deeper groundwater. Here, the anatomy of S. asiatica, including that of the axes and rootlets, is reported in detail for the first time. The axis contains stelar tissues, including pith, primary and secondary xylem, and phloem, and both primary and secondary cortical tissues. The primary cortex is tripartite and is divided into inner, middle, and outer zones. Secondary cortex is produced in the interior of the outer cortex splitting the outer cortex into inner and outer portions. The homogeneous pith and mesarch primary xylem maturation of S. asiatica are different from previously recorded species of Stigmaria Brongniart. Rootlets are composed of a central monarch vascular bundle surrounded by inner cortex and a ring of outer cortex. A vacant region is present between the inner and outer cortices. In some cases, connectives between the inner and outer cortices are present. According to the presented statistical analysis, it is proposed that rootlets of S. asiatica were highly branched, with at least for 7 bifurcations. Rootlets, together with the downward penetrating rhizomorph, helped to both anchor the plant and search for and absorb deep groundwater.

Despite the known hazards to wildlife, plastic waste to waterways and oceans continues to increase at alarming rates every year. Nanoplastics are a breakdown product of plastic debris in the environment (0.1–100 nm). Their small size renders them bioavailable, and they are likely to accumulate/adsorb persistent organic pollutants, which could increase their toxicity. In this study, we evaluated the effects of nanoplastics and phenanthrene on the early life stages of zebrafish. Zebrafish embryos were exposed to fluorescent nanoparticles and observed with fluorescence microscopy. Polystyrene nanoplastics (80 nm) attached to the embryonic chorionic membrane and were ingested when zebrafish reached 144 h after fertilization, the point at which they first open their mouth, as observed by the fluorescence in their intestines. Exposure alone or in combination with phenanthrene resulted in zebrafish pericardial edema, spinal curvature, increased embryonic heart rate, shortened body length, and increased mortality. Co-exposure to 5 mg/L nanoplastics and 0.1 mg/L phenanthrene caused more negative effects on length, teratogenicity and mortality of zebrafish embryos than the corresponding concentrations of single exposure. This study provides clear evidence that nanoplastics can be ingested by zebrafish larvae, thus future work to assess the toxicological effects is important to marine wildlife management.

Edible insects, regarded as a potential contributor to food security are currently given wide consideration due to their rich protein and other micronutrients contents. In this study, protease-assisted hydrolysis proposes an economically effective approach to hydrolyse proteins from silkworm (Bombyx mori) pupae to improve its functional properties. The proteolytic activity of a novel neutral protease (265.14 U/ml) with appreciable thermal activities, was identified using 16S rDNA as Stenotrophomonas maltophilia JW20 (SmNP20). The neutral protease with an apparent molecular weight of 28 kDa emerged active at pH 7 and maintained stability in pH range 6.0-8.0. The optimum temperature was 60 °C and stable at 55-60 °C, maintaining over 80% of its initial activity, with a half-life of 78.75, 89, 66.8 and 44 min at 50, 60, 70 and 80 °C. It was purified to 9.98-fold with a specific activity of 455.06 U/mg and 63.73% yield. The Km and Vmax values were 0.70 mg/ml and 9.48 μmol/min/mg, respectively. Enzymolysis with neutral protease enhanced the degree of hydrolysis (97.46±4.87%), increased water solubility over 50%, and a significant protein solubility of 63.44±0.65%. The Km and Vmax of the protein yield were 0.24 mg/ml and 165.63 μmol/min/mg respectively. A total of 17 amino acids have been detected in the hydrolysates obtained from the silkworm pupae protein. In comparison with neutrase and flavorzyme®, the enzyme possesses an elevated hydrolytic and catalytic efficiency. Emulsion activity and foam capacity ranged from 8-48 m2/g and 6-25% respectively. Hence, this study confirms the unique and efficient characteristics of an insect-enzyme correlation that is practically significant with potential improvement in nutritional composition and functional quality of insect proteins.

SummaryGlnR activates nitrogen metabolism genes under nitrogen-limited conditions whereas MtrA represses these genes under nutrient-rich conditions in Streptomyces. In this study, we compared the transcription patterns of nitrogen metabolism genes in a double deletion mutant (ΔmtrA-glnR) lacking both mtrA and glnR and in mutants lacking either mtrA (ΔmtrA) or glnR (ΔglnR). The nitrogen metabolism genes were expressed similarly in ΔmtrA-glnR and ΔglnR under both nitrogen-limited and nutrient-rich conditions, with patterns distinctly different from that of ΔmtrA, suggesting a decisive role for GlnR in the control of nitrogen metabolism genes and further suggesting that regulation of these genes by MtrA is GlnR-dependent. MtrA and GlnR utilize the same binding sites upstream of nitrogen metabolism genes, and we showed stronger in vivo binding of MtrA to these sites under nutrient-rich conditions and of GlnR under nitrogen-limited conditions, consistent with the higher levels of MtrA or GlnR under those respective conditions. In addition, we showed that both mtrA and glnR are auto-regulatory. Our study provides new insights into the regulation of nitrogen metabolism genes in Streptomyces.

We measured the concentrations of Cr, Fe, Ni, Cu, Zn, Cd, Pb and Hg, and the stable isotope ratios (δ13C and δ15N) in 87 fish samples within 12 economic fish species collected from the Dongting Lake, the second largest freshwater lake in China. With few exceptions in concentration of Cr, most of fish species showed lower concentrations of the 8 metals than legislation thresholds. Piscivorous fishes had significantly higher values of δ15N (possessing higher trophic level) and metal concentrations than planktivorous and herbivorous fishes. Moreover, demersal fishes showed higher concentrations of Cu, Zn, Cd and Pb than pelagic and benthopelagic fishes. We found positive correlations between concentrations of Fe, Pb and Hg and δ15N ratio, confirming the biomagnification of the three metals through trophic transfer. In contrast, fishes showed clear growth dilution effect for Ni, Zn, Cu, Pb and Hg, indicated by the negative correlations between their concentrations and fish weight or length. Multiple regression analysis demonstrated that growth dilution and biomagnification effects simultaneously governed the metal concentrations in fish muscle, and the two effects' importance varied among different metals. The human health risk assessment indicated that all 8 metals gave target hazard quotient (THQ) values

Previous study demonstrated that Ganoderma meroterpene derivative (GMD) increased the abundance of butyrate-producing bacteria in gut and subsequently delivered anti-metabolic disorder effect of host. To specify the key commensal bacteria associating with the beneficial effects, we tried to isolate and compare the microbiota from the cecal samples of GMD- and vehicle-treated ob/ob mice, and further identified butyrate-producing bacterial strains. It was found that Faeciroseburia intestinalis was enriched and 11 strains affiliated to F. intestinalis were cultivated from the gut of GMD-treated mice. In vitro assay attested butyrate production by representative strain of F. intestinalis. Oral administration with F. intestinalis further demonstrated its benefits on regulating hyperglycemia and hyperlipidemia, on decreasing plasma lipopolysaccharide (LPS) and inflammation, and on improving hepatic injuries. Treatment with F. intestinalis effectively enhanced the level of gut butyrate, which subsequently ameliorated the intestinal barrier function and activated epithelial PPAR-γ signaling pathway to regulate microbiome homeostasis in gut. Our study demonstrated that the causal relationship between the butyrate-producing bacteria and the GMD's therapeutic effects and confirmed the important function of the butyrate-producing F. intestinalis in maintaining host metabolism homeostasis.

Cryptocaryon irritans, a holotrichous ciliate parasitic protozoan, can trigger marine white spot disease and cause substantial economic losses in mariculture. However, methods of preventing and curing the disease have negatively affect fish, human, other organisms, and the natural environment. The antiparasitic activity of some antimicrobial peptides (AMPs) has garnered extensive attention of scholars. In this study, we identified and characterised a novel antiparasitic peptide, named So-pis, from Sciaenops ocellatus. The sequence analysis, structural features, and tissue distribution suggested that So-pis is genetically related to the piscidins family. However, So-pis showed a relatively low overall conservation compared with other known piscidins. So-pis is abound in glycine residues (22.7 %) and it has a neutral isoelectric point, weak amphipathicity, relatively long α-helix, and high hydrophobicity. These key elements are responsible for its biological activity. Quantitative real-time polymerase chain reaction (qRT-PCR) data indicated that So-pis is a typically gill-expressed peptide. The expression of So-pis in the gill, skin, spleen, and head kidney could be regulated during C. irritans infection, thereby implicating a role of So-pis in immune defence against C. irritans. The synthetic So-pis had limited or no antimicrobial activity against bacterial and yeasts but exhibited potent antiparasitic activity against C. irritans in vitro. The activity of synthetic So-pis against erythrocytes was less potent than its antiparasitic activity against C. irritans. These results indicated that So-pis might be one of the crucial defence cytokines against C. irritans in the red drum. Cumulatively, our data suggested that So-pis might be a potential candidate for developing a novel, effective, and safe therapeutic agent against marine white spot disease.

Micro/mesoporous powder structures and biofouling of marine microorganisms directly limit the potential of zeolite frameworks in uranium recovery from seawater. Herein, an anti-biofouling zeolite framework-8/chitosan/melamine sponge adsorbent with a three-dimensional porous structure was prepared successfully using zinc ions and chitosan as adjuvants to promote the growth of zeolite framework-8 on the surface of melamine sponge. The mechanical and anti-biofouling properties of the sponge were determined by compression and algae tests. Relative to the zeolite framework-8 powders, the zeolite framework-8/chitosan/melamine sponge exhibited superior mechanical and anti-biofouling properties with 0.565 MPa of compressive stress and a 35% algae cell death rate. The adsorption capacities of the zeolite framework-8/chitosan/melamine sponge were 129.90 mg g−1 at pH 8.0 and 95.47 mg g−1 at the fifth adsorption–desorption cycle, which were higher than that of ZIF-8 powder (91.45 mg g−1). Furthermore, the excellent low-concentration U(VI) adsorption performance of zeolite framework-8/chitosan/melamine in natural seawater (Bohai) also confirmed its practical application potentiality.

The influenza A M2 wild-type (WT) proton channel is the target of the anti-influenza drug rimantadine. Rimantadine has two enantiomers, though most investigations into drug binding and inhibition have used a racemic mixture. Solid-state NMR experiments using the full length-M2 WT have shown significant spectral differences that were interpreted to indicate tighter binding for (R)- vs (S)-rimantadine. However, it was unclear if this correlates with a functional difference in drug binding and inhibition. Using X-ray crystallography, we have determined that both (R)- and (S)-rimantadine bind to the M2 WT pore with slight differences in the hydration of each enantiomer. However, this does not result in a difference in potency or binding kinetics, as shown by similar values for kon, koff, and Kd in electrophysiological assays and for EC50 values in cellular assays. We concluded that the slight differences in hydration for the (R)- and (S)-rimantadine enantiomers are not relevant to drug binding or channel inhibition. To further explore the effect of the hydration of the M2 pore on binding affinity, the water structure was evaluated by grand canonical ensemble molecular dynamics simulations as a function of the chemical potential of the water. Initially, the two layers of ordered water molecules between the bound drug and the channel’s gating His37 residues mask the drug’s chirality. As the chemical potential becomes more unfavorable, the drug translocates down to the lower water layer, and the interaction becomes more sensitive to chirality. These studies suggest the feasibility of displacing the upper water layer and specifically recognizing the lower water layers in novel drugs.

Gene expression profiling has long been used in understanding the contribution of genes and related pathways in disease pathogenesis and susceptibility. We have performed whole-blood transcriptomic profiling in a subset of patients with inherited bone marrow failure (IBMF) whose diseases are clinically and genetically characterized as Fanconi anemia (FA), Shwachman-Diamond syndrome (SDS), and dyskeratosis congenita (DC). We hypothesized that annotating whole-blood transcripts genome wide will aid in understanding the complexity of gene regulation across these IBMF subtypes. Initial analysis of these blood-derived transcriptomes revealed significant skewing toward upregulated genes in patients with FA when compared with controls. Patients with SDS or DC also showed similar skewing profiles in their transcriptional status revealing a common pattern across these different IBMF subtypes. Gene set enrichment analysis revealed shared pathways involved in protein translation and elongation (ribosome constituents), RNA metabolism (nonsense-mediated decay), and mitochondrial function (electron transport chain). We further identified a discovery set of 26 upregulated genes at stringent cutoff (false discovery rate < 0.05) that appeared as a unified signature across the IBMF subtypes. Subsequent transcriptomic analysis on genetically uncharacterized patients with BMF revealed a striking overlap of genes, including 22 from the discovery set, which indicates a unified transcriptional drive across the classic (FA, SDS, and DC) and uncharacterized BMF subtypes. This study has relevance in disease pathogenesis, for example, in explaining the features (including the BMF) common to all patients with IBMF and suggests harnessing this transcriptional signature for patient benefit.

The main protease (Mpro) is a validated antiviral drug target of SARS-CoV-2. A number of Mpro inhibitors have now advanced to animal model study and human clinical trials. However, one issue yet to be addressed is the target selectivity over host proteases such as cathepsin L. In this study we describe the rational design of covalent SARS-CoV-2 Mpro inhibitors with novel cysteine reactive warheads including dichloroacetamide, dibromoacetamide, tribromoacetamide, 2-bromo-2,2-dichloroacetamide, and 2-chloro-2,2-dibromoacetamide. The promising lead candidates Jun9-62-2R (dichloroacetamide) and Jun9-88-6R (tribromoacetamide) had not only potent enzymatic inhibition and antiviral activity but also significantly improved target specificity over caplain and cathepsins. Compared to GC-376, these new compounds did not inhibit the host cysteine proteases including calpain I, cathepsin B, cathepsin K, cathepsin L, and caspase-3. To the best of our knowledge, they are among the most selective covalent Mpro inhibitors reported thus far. The cocrystal structures of SARS-CoV-2 Mpro with Jun9-62-2R and Jun9-57-3R reaffirmed our design hypothesis, showing that both compounds form a covalent adduct with the catalytic C145. Overall, these novel compounds represent valuable chemical probes for target validation and drug candidates for further development as SARS-CoV-2 antivirals.

Three undescribed dammarane-type triterpenoids with rare carbon skeletons (1-3), together with one unreported gypenoside (4), were isolated from a acid hydrolysis product of Gynostemma pentaphyllum (Thunb.) Makino. Their molecular structures were elucidated via a combination of spectroscopic and spectrometric data, while the absolute configurations of compounds 1, 2 and 4 were determined by electronic circular dichroism (ECD) analyse. Notably, compounds 1-3 possess a 19(10→1)abeo structural moiety in the A ring. Saponin 4 possesses an unusual cyclopentenone side chain. Compounds 1, 2 and 4 were screened for their antiproliferative effects against HepG2, MCF-7 and DU145 cell lines.

Background: Anopheles gambiae (An. gambiae) is considered the most effective malaria vector worldwide and is widely distributed in Africa. The purpose of this study is to determine the potential impact of climate change on An. gambiae and predict the present and future potential suitable habitat globally. Methods: In this study, environmental variables, global occurrence data of An. gambiae, and the maximum entropy model (MaxEnt) were used to evaluate the contribution of environmental factors and predict the habitat suitability of An. gambiae under the current and future scenarios. Results: Among all environmental variables, isothermality (Bio3, 34.5%) contributed the most to An. gambiae distribution. Under current climate conditions, the potential suitable areas for An. gambiae are mainly located near the equator (approximately 30°N-30°S), with a total area of 16.58 million km2, including central and northern South America, a fraction of areas near the equator of North America, central and southern Africa, some tropical regions of southern Asia, and small areas of Oceania. The areas of potential suitable habitats would be reduced to varying degrees in future climate scenarios. Conclusions: Potential suitable habitats for An. gambiae may not be limited to Africa. Necessary surveillance and preventive measures should be undertaken in high-risk regions, including those outside Africa, to monitor and control the spread of An. gambiae.

Soil priming is a microbial-driven process, which determines key soil–climate feedbacks in response to fresh carbon inputs. Despite its importance, the microbial traits behind this process are largely undetermined. Knowledge of the role of these traits is integral to advance our understanding of how soil microbes regulate carbon (C) emissions in forests, which support the largest soil carbon stocks globally. Using metagenomic sequencing and C-glucose, we provide unprecedented evidence that microbial traits explain a unique portion of the variation in soil priming across forest biomes from tropical to cold temperature regions. We show that microbial functional profiles associated with the degradation of labile C, especially rapid simple sugar metabolism, drive soil priming in different forests. Genes involved in the degradation of lignin and aromatic compounds were negatively associated with priming effects in temperate forests, whereas the highest level of soil priming was associated with β-glucosidase genes in tropical/subtropical forests. Moreover, we reconstructed, for the first time, 42 whole bacterial genomes associated with the soil priming effect and found that these organisms support important gene machinery involved in priming effect. Collectively, our work demonstrates the importance of microbial traits to explain soil priming across forest biomes and suggests that rapid carbon metabolism is responsible for priming effects in forests. This knowledge is important because it advances our understanding on the microbial mechanisms mediating soil–climate feedbacks at a continental scale., This work were financially supported by the National Natural Science Foundation of China (41907031), the Chinese Academy of Sciences “Light of West China” Program for Introduced Talent in the West, the National Natural Science Foundation of China (31570440, 31270484), the Key International Scientific and Technological Cooperation and Exchange Project of Shaanxi Province, China (2020KWZ-010), the 2021 First Funds for Central Government to Guide Local Science and Technology Development in Qinghai Province (2021ZY002), the i-LINK +2018 (LINKA20069) from CSIC, and a Ramón y Cajal grant from the Spanish Ministry of Science and Innovation (RYC2018-025483-I)

Biocompatible photo-driven producers of singlet oxygen can inhibit the growth of drug-resistant bacteria and tumors. In order to develop bacteria targeting generator of singlet oxygen for tumor and bacterial treatment, a metal porphyrin liposome (Phy–Mn–Ls) was prepared by the metal coordination reaction and self-assembly of porphyrin compounds with bacteria targeting polymer (HS–PEG–chol). The photo-driven production of 1O2, binding with bovine serum protein (BSA) and lipase, toxicity to MCF-7 breast cancer cells and inhibitory effect on the growth of Escherichia coli have been investigated. Fluorescence analysis results show that Phy–Mn–Ls can bind to lipase, and it shows less effect on the conformation of BSA and is low cytotoxicity without irradiation. In particular, the good biocompatibility made Phy–Mn–Ls exhibit good photosensitive antibacterial activity and anti-tumor properties. The results demonstrate that the coordination of HS–PEG–chol with metal-phorphrin coodination is an effective way to develop bacteria targeting nano-complexes (Phy–Mn–Ls) for lipase affinity and photodriven bacteria treatment.

Chronic visceral pain (CVP) is one of the common symptoms of many diseases triggered by underlying diseases of the internal organs of the human body. Its causes include vascular mechanisms, mechanical factors, persistent inflammation, and unexplained functional mechanisms. Although the pathogenesis is unclear, more and more research has begun to shift from the neuronal aspect to the glial cells in recent years. Some data highlight that the spinal glial cells, particularly the microglia and astrocytes, play an essential role in CVP. Based on this, we highlight the mechanisms of microglia and astrocytes in CVP concerning the release of cytokines, chemokines, and neuroactive substances and alterations in intracellular signaling pathways during the process. Finally, because CVP is widespread in various diseases, we present future perspectives targeting microglia and astrocytes for treatment.

Introduction The experimental cerebral malaria (ECM) model in C57BL/6 mice infected with Plasmodium berghei ANKA (PbA) has revealed microglia are involved in the ECM immune microenvironment. However, the regulation of microglia in the ECM immune response is not clear, and there is no safe and efficient treatment clinically for the protection of the nerve cells. Aims To elucidate the negative regulation mechanism in the ECM brain mediated by microglia. Furthermore, to investigate protective effect of the appropriate enhancement of the PD‐1/PD‐L1 pathway in the brain against ECM through the intrathecal injection of the adenovirus expressing PDL1‐IgG1Fc fusion protein. Results The PD‐1/PD‐L1 pathway was induced in the ECM brain and showed an upregulation in the microglia. Deep single‐cell analysis of immune niches in the ECM brainstem indicated that the microglia showed obvious heterogeneity and activation characteristics. Intrathecal injection of recombinant adenovirus expressing PD‐L1 repressed the neuroinflammation and alleviated ECM symptoms. In addition, the synergistic effect of artemisinin and intracranial immunosuppression mediated by PD‐L1 was more efficacious than either treatment alone. Conclusion The appropriate enhancement of the PD‐1/PD‐L1 pathway in the early stage of ECM has an obvious protective effect on the maintenance of immune microenvironment homeostasis in the brain. Regulating microglia and the PD‐1/PD‐L1 pathway could be considered as a promising approach for protection against human cerebral malaria in the future., Microglia activated and expressed PD‐L1 and PD‐1 induced by PbA infection.The PD‐L1 could interacted with PD‐1 on the infiltrated CTL cells and inhibited the killing activity of CTL cells.The intrathecal injection of expression vector PDL1‐IgGFc could inhibit intracerebral inflammation and improved ECM.

Understanding the spatial structure of genetic diversity provides insights into a populations' genetic status and enables assessment of its capacity to counteract the effects of genetic drift. Such knowledge is particularly scarce for the snow leopard, a conservation flagship species of Central Asia mountains. Focusing on a snow leopard population in the Qilian mountains of Gansu Province, China, we characterised the spatial genetic patterns by incorporating spatially explicit indices of diversity and multivariate analyses, based on different inertia levels of Principal Component Analysis (PCA). We compared two datasets differing in the number of loci and individuals. We found that genetic patterns were significantly spatially structured and were characterised by a broad geographical division coupled with a fine-scale cline of differentiation. Genetic admixture was detected in two adjoining core areas characterised by higher effective population size and allelic diversity, compared to peripheral localities. The power to detect significant spatial relationships depended primarily on the number of loci, and secondarily on the number of PCA axes. Spatial patterns and indices of diversity highlighted the cryptic structure of snow leopard genetic diversity, likely driven by its ability to disperse over large distances. In combination, the species' low allelic richness and large dispersal ability result in weak genetic differentiation related to major geographical features and isolation by distance. This study illustrates how cryptic genetic patterns can be investigated and analysed at a fine spatial scale, providing insights into the spatially variable isolation effects of both geographic distance and landscape resistance.

Yin Tang,1,2,&ast; Yu Cui,3– 5,&ast; Lin-lin Li,6,&ast; Ya-ping Guan,3– 5 Dong-feng Feng,3– 5 Bei-bei Yin,3– 5 Xue-feng Liang,2 Jing Yin,2 Rui Jiang,6 Jing Liang,3– 5 Ya-hong Sun,2 Jun Wang3– 5 1Postgraduate Department, Shandong First Medical University & Shandong Academy of Medical Sciences, Jinan, 250117, People’s Republic of China; 2Department of Oncology, The Third Affiliated Hospital of Shandong First Medical University (Affiliated Hospital of Shandong Academy of Medical Sciences), Jinan, 250031, People’s Republic of China; 3Department of Oncology, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Jinan, 250014, People’s Republic of China; 4Shandong Lung Cancer Institute, Jinan, 250014, People’s Republic of China; 5Shandong Key Laboratory of Rheumatic Disease and Translational Medicine, Jinan, 250014, People’s Republic of China; 6Department of Oncology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, 250021, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ya-hong SunDepartment of Oncology, The Third Affiliated Hospital of Shandong First Medical University (Affiliated Hospital of Shandong Academy of Medical Sciences), No. 38, Wu Yingshan Road, Tianqiao District, Jinan, 250031, People’s Republic of ChinaTel +86-531-58628551Email sunyahong@sdfmu.edu.cnJun WangDepartment of Oncology, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, No. 16766, Jingshi Road, Lixia District, Jinan, 250014, People’s Republic of ChinaTel +86-531-89268839Email ggjun2005@126.comPurpose: To evaluate the dynamics of early serum tumour markers (STMs) and the neutrophil-to-lymphocyte ratio (NLR) to predict clinical efficacy and prognosis of advanced non-small-cell lung cancer (NSCLC) patients who received programmed cell death-1 (PD-1)/programmed cell death-ligand 1 (PD-L1) inhibitors.Patients and Methods: We retrospectively reviewed patients with advanced NSCLC treated with PD-1/PD-L1 inhibitors between September 2017 and August 2020. NLR and STMs were routinely measured between immunotherapy initiation and the first radiological evaluation. A combination score based on the leading STM and NLR and their dynamic changes was established. The effects of leading STM change, NLR change, and the combination score on the objective response rate (ORR), durable clinical benefit (DCB), progression-free survival (PFS), and overall survival (OS) were analysed. The accuracy of the combination score was evaluated by receiver operating characteristic (ROC) curve and the area under the curve (AUC).Results: Overall, 124 patients were included in this retrospective cohort study. The ORR was 22.8%, DCB was 54.5%, and the median OS and PFS were 21.6 and 14.9 months, respectively. Patients with low combination scores had a significantly improved ORR and DCB compared with those with intermediate or high scores (P = 0.002 for ORR, P < 0.0001 for DCB). In a multivariate model, the combination score was an independent indicator of PFS (P < 0.0001) and OS (P < 0.0001). The AUC demonstrated that the combination score (AUC = 0.706) has greater predictive power than either the posttreatment NLR (AUC = 0.668) or the leading STM change (AUC = 0.648) alone.Conclusion: An easy, cost-effective, and novel combination score based on the dynamics of an early STM and the NLR can accurately predict the clinical efficacy of PD-1/PD-L1 inhibitors and prognosis in advanced NSCLC patients.Keywords: serum tumour marker, neutrophil-to-lymphocyte ratio, PD-1 inhibitors, non-small-cell lung cancer, prognosis

Fresh Milk Bar (FMB), an emerging dairy retail franchise, is used to instantly produce and sell pasteurized milk and other dairy products in China. However, the quality and safety of pasteurized milk in FMB have received little attention. The objective of this study was to investigate the prevalence, antimicrobial resistance, and virulence genes of Escherichia coli, Staphylococcus aureus, and Streptococcus in 205 pasteurized milk samples collected from FMBs in China. Four (2.0%) isolates of E. coli, seven (3.4%) isolates of S. aureus, and three (1.5%) isolates of Streptococcus agalactiae were isolated and identified. The E. coli isolates were resistant to amikacin (100%), streptomycin (50%), and tetracycline (50%). Their detected resistance genes include aac(3)-III (75%), blaTEM (25%), aadA (25%), aac(3)-II (25%), catI (25%), and qnrB (25%). The S. aureus isolates were mainly resistant to penicillin G (71.4%), trimethoprim-sulfamethoxazole (71.4%), kanamycin (57.1%), gentamicin (57.1%), amikacin (57.1%), and clindamycin (57.1%). blaZ (42.9%), mecA (28.6%), ermB (14.3%), and ermC (14.3%) were detected as their resistance genes. The Streptococcus strains were mainly resistant to tetracycline (66.7%) and contained the resistance genes pbp2b (33.3%) and tetM (33.3%). The virulence genes eae and stx2 were only found in one E. coli strain (25%), sec was detected in two S. aureus strains (28.6%), and bca was detected in one S. agalactiae strain (33.3%). The results of this study indicate that bacteria with drug resistance and virulence genes isolated from the pasteurized milk of FMB are a potential risk to consumers' health.

Most previous studies in the pathophysiology of major depressive disorder (MDD) focused on fecal samples, which limit the identification of the gut mucosal and luminal microbiome in depression. Here, we address this knowledge gap. Male cynomolgus macaques (Macaca fascicularis) were randomly assigned to a chronic unpredictable mild stress (CUMS) group, or to an unstressed control group. Behavioral tests were completed in both groups. At endpoint, microbe composition of paired mucosal and luminal samples from cecum, ascending, transverse, and descending colons were determined by 16S ribosomal RNA gene sequencing. The levels of 34 metabolites involved in carbohydrate or energy metabolism in luminal samples were measured by targeted metabolomics profiling. CUMS macaques demonstrated significantly more depressive-like behaviors than controls. We found differences in mucosal and luminal microbial composition between the two groups, which were characterized by Firmicutes and Bacteriodetes at the phylum level, as well as Prevotellaceae and Lachnospiraceae at the family level. The majority of discriminative microbes correlated with the depressive-like behavioral phenotype. In addition, we found 27 significantly different microbiome community functions between the two groups in mucosa, and one in lumen, which were mainly involved in carbohydrate and energy metabolism. A total of nine metabolites involved in these pathways were depleted in CUMS animals. Together, CUMS macaques with depressive-like behaviors associated with distinct alterations of covarying microbiota, carbohydrate and energy metabolism in mucosa and lumen. Further studies should focus on the mucosal and luminal microbiome to provide a deeper spatiotemporal perspective of microbial alterations in the pathogenesis of MDD.

Long non-coding RNAs (lncRNAs) act as important regulators of tumorigenesis and development in bladder cancer. However, the underlying molecular mechanisms remain elusive. We previously identified a novel lncRNA signature related to immunity and progression in bladder cancer. Here we further explored the function of RP11-89, a lncRNA discovered in the previous signature. Loss- and gain-of function experiments were performed using CCK-8 assay, flow cytometry, Transwell assays, scratch tests and subcutaneous nude mouse models. High-throughput RNA sequencing was conducted to identify dysregulated genes in bladder cancer cells with RP11-89 knockdown or overexpression. Regulation of RP11-89 on miR-129-5p and PROM2 was explored through luciferase reporter assay, RIP assay and RNA pull-down assay. RP11-89 promoted cell proliferation, migration and tumorigenesis and inhibited cell cycle arrest via the miR-129-5p/PROM2 axis. We found that RP11-89 “sponges” miR-129-5p and upregulates PROM2. Elevated PROM2 in cells was associated with attenuated ferroptosis through iron export, formation of multivesicular bodies and less mitochondrial abnormalities. We demonstrated that RP11-89 is a novel tumorigenic regulator that inhibits ferroptosis via PROM2-activated iron export. RP11-89 may serve as a potential biomarker for targeted therapy in bladder cancer.

Carotenoids are involved in many essential physiological functions and are produced from geranylgeranyl pyrophosphate through synthase, desaturase, and cyclase activities. In the pea aphid (Acyrthosiphon pisum), the duplication of carotenoid biosynthetic genes, including carotenoid synthases/cyclases (ApCscA-C) and desaturases (ApCdeA-D), through horizontal gene transfer from fungi has been detected, and ApCdeB has known dehydrogenation functions. However, whether other genes contribute to aphid carotenoid biosynthesis, and its specific regulatory pathway, remains unclear. In the current study, functional analyses of seven genes were performed using heterologous complementation and RNA interference assays. The bifunctional enzymes ApCscA-C were responsible for the synthase of phytoene, and ApCscC may also have a cyclase activity. ApCdeA, ApCdeC, and ApCdeD had diverse dehydrogenation functions. ApCdeA catalyzed the enzymatic conversion of phytoene to neurosporene (three-step product), ApCdeC catalyzed the enzymatic conversion of phytoene to ζ-carotene (two-step product), and ApCdeD catalyzed the enzymatic conversion of phytoene to lycopene (four-step product). Silencing of ApCscs reduced the expression levels of ApCdes, and silencing these carotenoid biosynthetic genes reduced the α-, β-, and γ-carotene levels, as well as the total carotenoid level. The results suggest that these genes were activated and led to carotenoid biosynthesis in the pea aphid.

Objectives Nuclear factor I-C (NFIC) plays a critical role in regulating epithelial-mesenchymal crosstalk during tooth development. However, it remains largely unknown about how NFIC functions in dentin and enamel formation. In the present review, we aim to summarize the most recent discoveries in the field and gain a better understanding of the roles NFIC performs during tooth formation. Subjects and methods Nfic-/- mice exhibit human dentin dysplasia type I (DDI)-like phenotypes signified by enlarged pulp chambers, the presence of short-root anomaly, and failure of odontoblast differentiation. Although loss of NFIC has little effect on molar crown morphology, researchers have detected aberrant microstructures of enamel in the incisors. Recently, accumulating evidence has further uncovered the novel function of NFIC in the process of enamel and dentin formation. Results During epithelial-mesenchyme crosstalk, the expression of NFIC is under the control of SHH-PTCH-SMO-GLI1 pathway. NFIC is closely involved in odontoblast lineage cells proliferation and differentiation, and the maintenance of NFIC protein level in cytoplasm is negatively regulated by TGF-β signaling pathway. In addition, NFIC has mild effect on ameloblast differentiation, enamel mineralization and cementum formation. Conclusions NFIC plays an important role in tooth development and is required for the formation of dentin, enamel as well as cementum.

Background Xylanase, an important accessory enzyme that acts in synergy with cellulase, is widely used to degrade lignocellulosic biomass. Thermostable enzymes with good catalytic activity at lower temperatures have great potential for future applications in the feed and fuel industries, which have distinct demands; however, the potential of the enzymes is yet to be researched. Results In this study, a structure-based semi-rational design strategy was applied to enhance the low-temperature catalytic performance of Bispora sp. MEY-1 XYL10C_∆N wild-type (WT). Screening and comparisons were performed for the WT and mutant strains. Compared to the WT, the mutant M53S/F54L/N207G exhibited higher specific activity (2.9-fold; 2090 vs. 710 U/mg) and catalytic efficiency (2.8-fold; 1530 vs. 550 mL/s mg) at 40 °C, and also showed higher thermostability (the melting temperature and temperature of 50% activity loss after 30 min treatment increased by 7.7 °C and 3.5 °C, respectively). Compared with the cellulase-only treatment, combined treatment with M53S/F54L/N207G and cellulase increased the reducing sugar contents from corn stalk, wheat bran, and corn cob by 1.6-, 1.2-, and 1.4-folds, with 1.9, 1.2, and 1.6 as the highest degrees of synergy, respectively. Conclusions This study provides useful insights into the underlying mechanism and methods of xylanase modification for industrial utilization. We identified loop2 as a key functional area affecting the low-temperature catalytic efficiency of GH10 xylanase. The thermostable mutant M53S/F54L/N207G was selected for the highest low-temperature catalytic efficiency and reducing sugar yield in synergy with cellulase in the degradation of different types of lignocellulosic biomass. Graphic Abstract Supplementary Information The online version contains supplementary material available at 10.1186/s13068-021-02044-3., Highlights Key sites were proved to promote low-temperature catalytic efficiency of xylanase.Thermostable mutant with improved catalytic efficiency at 40 °C was obtained.Synergistic hydrolysis was dramatically improved between the mutant and cellulase.Agricultural wastes were converted into fermentable sugars by enzymolysis. Supplementary Information The online version contains supplementary material available at 10.1186/s13068-021-02044-3.

Friedreich’s ataxia (FRDA) is a severe multisystem disease caused by transcriptional repression induced by expanded GAA repeats located in intron 1 of the Frataxin (FXN) gene encoding frataxin. FRDA results from decreased levels of frataxin; thus, stabilization of the FXN mRNA already present in patient cells represents an attractive and unexplored therapeutic avenue. In this work, we pursued a novel approach based on oligonucleotide-mediated targeting of FXN mRNA ends to extend its half-life and availability as a template for translation. We demonstrated that oligonucleotides designed to bind to FXN 5′ or 3′ noncoding regions can increase FXN mRNA and protein levels. Simultaneous delivery of oligonucleotides targeting both ends increases efficacy of the treatment. The approach was confirmed in several FRDA fibroblast and induced pluripotent stem cell-derived neuronal progenitor lines. RNA sequencing and single-cell expression analyses confirmed oligonucleotide-mediated FXN mRNA upregulation. Mechanistically, a significant elongation of the FXN mRNA half-life without any changes in chromatin status at the FXN gene was observed upon treatment with end-targeting oligonucleotides, indicating that transcript stabilization is responsible for frataxin upregulation. These results identify a novel approach toward upregulation of steady-state mRNA levels via oligonucleotide-mediated end targeting that may be of significance to any condition resulting from transcription downregulation.

Purpose Mitochondria are closely correlated with the proliferation and metastasis of tumor for providing suitable micro-environment and energy supply. Herein, we construct a glucose transporter 1 (GLUT1) targeting and hypoxia activating polyprodrug-based micelle (Glu-PEG-Azo-IR808-S-S-PTX) for mitochondria-specific drug delivery and tumor chemo-thermal therapy. Results The micelle was characterized by hypoxia-sensitive PEG outer layer detachment, high photo-thermal conversion efficiency, and glutathione (GSH)-sensitive paclitaxel （PTX） release. It showed GLUT1 specifically cellular uptake and hypoxia-sensitive mitochondria targeting on A549 cell. In vivo fluorescence imaging confirmed the micelle also could selectively accumulate in tumor and its mitochondria on A549 tumor-bearing nude mice. Consequently, it not only exhibited higher cytotoxicity, apoptosis rate, and metastasis inhibition rate on A549 cells, but also better tumor growth and metastasis inhibition rate on tumor-bearing nude mice and lower whole-body toxicity. The mechanism might be caused by destroying mitochondria and down-regulating ATP production. Conclusion This study provided a GLUT1 targeting, hypoxia, and reductive responsive nanomedicine that hold the potential to be exploited for tumor therapy.

The inverse correlation between high-density lipoprotein (HDL) levels in vivo and the risk of Alzheimer's disease (AD) has become an inspiration for HDL-inspired AD therapy, including plain HDL and various intelligent HDL-based drug delivery systems. In this review, we will focus on the two endogenous HDL subtypes in the central nervous system (CNS), apolipoprotein E-based HDL (apoE-HDL) and apolipoprotein A-I-based HDL (apoA-I-HDL), especially their influence on AD pathophysiology to reveal HDL's potential as biomarkers for risk prediction, and summarize the relevant therapeutic mechanisms to propose possible treatment strategies. We will emphasize the latest advances of HDL as therapeutics (plain HDL and HDL-based drug delivery systems) to discuss the potential for AD therapy and review innovative techniques in the preparation of HDL-based nanoplatforms to provide a basis for the rational design and future development of anti-AD drugs.

Background Tanning is an important physiological process that play the critical roles in affecting cuticle pigmentation and sclerotization. Previous studies have showed that insect cuticle tanning was closely associated with tyrosine metabolism pathway that consisted of a series of enzymes. Results In this study, a total of 24 tyrosine metabolism pathway genes were identified in the oriental fruit fly Bactrocera dorsalis (Hendel) genome. Then, the gene expression profiles throughout 15 developmental stages of B. dorsalis were established on the basis of our previous RNA-seq data, and we found 13 enzyme genes could be involved in the process of pupariation. Accordingly, a predicted tyrosine-mediated tanning pathway during the pupariation of B. dorsalis was obtained, and a critical enzyme 3,4-dihydroxyphenylalanine (DOPA) decarboxylase (DDC) was selected to explore its possible functions in the formation of puparium. Firstly, the RT-qPCR confirmed that BdDDC had epidermis-specific expression pattern, and it was also highly expressed during the larval metamorphosis of B. dorsalis. Subsequently, the dysfunction of BdDDC by feeding 5-day-old larvae with DDC inhibitor (L-α-Methyl-DOPA) could lead to (i) the significant decreasing of BdDDC enzyme activity and dopamine concentration; (ii) the defects in puparium pigmentation; (iii) the impairment of morphology and less thickness of puparium; (iv) the lower pupal weight and obstacle to eclosion. Conclusion The present study provided us a potential tyrosine metabolic pathway that was responsible for insect tanning during the pupariation, and the BdDDC enzyme has been proved to act the crucial roles in larval-pupal tanning of B. dorsalis. This article is protected by copyright. All rights reserved.

As a pancreatic inflammatory marker, regenerating islet-derived protein 3A (Reg3A) plays a key role in inflammation-associated pancreatic carcinogenesis by promoting cell proliferation, inhibiting apoptosis, and regulating cancer cell migration and invasion. This study aimed to reveal a novel immuno-regulatory mechanism by which Reg3A modulates tumour-promoting responses during pancreatic cancer (PC) progression. In an in vitro Transwell system that allowed the direct co-culture of human peripheral blood-derived dendritic cells (DCs) and Reg3A-overexpressing/ silenced human PC cells, PC cell-derived Reg3A was found to downregulate CD80, CD83 and CD86 expression on educated DCs, increase DC endocytic function, inhibit DC-induced T lymphocyte proliferation, reduce IL-12p70 production, and enhance IL-23 production by DCs. The positive effect of tumour-derived Reg3A-educated human DCs on PC progression was demonstrated in vivo by intraperitoneally transferring them into PC-implanted severe combined immunodeficiency (SCID) mice reconstituted with human T cells. A Reg3A-JAK2/STAT3 positive feedback loop was identified in DCs educated with Reg3A. In conclusion, as a tumour-derived factor, Reg3A acted to block the differentiation and maturation of the most important antigen-presenting cells, DCs, causing them to limit their potential anti-tumour responses, thus facilitating PC escape and progression.

Neoantigens play a key role in the recognition of tumour cells by T cells; however, only a small proportion of neoantigens truly elicit T-cell responses, and few clues exist as to which neoantigens are recognized by which T-cell receptors (TCRs). We built a transfer learning-based model named the pMHC–TCR binding prediction network (pMTnet) to predict TCR binding specificities of the neoantigens—and T cell antigens in general—presented by class I major histocompatibility complexes. pMTnet was comprehensively validated by a series of analyses and exhibited great advances over previous works. By applying pMTnet to human tumour genomics data, we discovered that neoantigens were generally more immunogenic than self-antigens, but human endogenous retrovirus E (a special type of self-antigen that is reactivated in kidney cancer) is more immunogenic than neoantigens. We further discovered that patients with more clonally expanded T cells that exhibit better affinity against truncal rather than subclonal neoantigens had more favourable prognosis and treatment response to immunotherapy in melanoma and lung cancer but not in kidney cancer. Predicting TCR–neoantigen/antigen pairing is one of the most daunting challenges in modern immunology; however, we achieved an accurate prediction of the pairing using only the TCR sequence (CDR3β), antigen sequence and class I major histocompatibility complex allele, and our work revealed unique insights into the interactions between TCRs and major histocompatibility complexes in human tumours, using pMTnet as a discovery tool. T-cell immunity is driven by the interaction between peptides presented by major histocompatibility complexes (pMHCs) and T-cell receptors (TCRs). Only a small proportion of neoantigens elicit T-cell responses, and it is not clear which neoantigens are recognized by which TCRs. The authors develop a transfer learning model to predict TCR binding specificity to class-I pMHCs.

Plants of genus Eleutherococcus belong to Araliaceae family, which mainly distributed in Asia. Several Eleutherococcus species have been used as ancient medicinal plants for many years in different traditional medical systems. The secondary metabolites of plants of genus Eleutherococcus have been studied since 1960s. In recent years, with the boom of "return to nature" and "the renewed emphasis on natural medicine", the investigations of genus Eleutherococcus have made new progress. So far, 422 secondary metabolites have been isolated and identified from 17 species. There are various pharmacological properties, such as anti-inflammation, anti-tumor, anti-anxiety, anti-depression, anti-diabetes, anti-fatigue, neuroprotection, hepatoprotection, immunoregulation, anti-Parkinson’s disease, anti-Alzheimer's disease, and anti-cardiovascular and cerebrovascular disease, most of which can be found internal connection with traditional medicine. Moreover, clinical researches have revealed the efficacy of drugs made from Eleutherococcus senticosus (Rupr. & Maxim.) Harms in the treatment of cardiovascular and cerebrovascular diseases, which have been marketed in China for decades. Despite a large number of literature reports regarding to the phytochemistry and biological activity of various Eleutherococcus species have been issued, there is not a complete and comprehensive review. Therefore, we discuss knowledge on ethnopharmacology, secondary metabolites, pharmacological and biological activity, clinical trial, and quality control of drugs made from plants of genus Eleutherococcus. Furthermore, deficiencies on present studies have also been discussed so as to provide reference for further research and promote the contemporary development of Eleutherococcus genus.

Background: Alzheimer’s disease (AD) is a degenerative disorder, accompanied by progressive cognitive decline, for which there is no cure. Recently, the close correlation between AD and type 2 diabetes mellitus (T2DM) has been noted, and a promising anti-AD strategy is the use of anti-T2DM drugs. Objective: To investigate if the novel glucagon-like peptide-1 (GLP-1)/glucose-dependent insulinotropic polypeptide (GIP) receptor agonist DA4-JC shows protective effects in the triple APP/PS1/tau mouse model of AD. Methods: A battery of behavioral tests were followed by in vivo recording of long-term potentiation (LTP) in the hippocampus, quantified synapses using the Golgi method, and biochemical analysis of biomarkers. Results: DA4-JC improved cognitive impairment in a range of tests and relieved pathological features of APP/PS1/tau mice, enhanced LTP in the hippocampus, increased numbers of synapses and dendritic spines, upregulating levels of post-synaptic density protein 95 (PSD95) and synaptophysin (SYP), normalized volume and numbers of mitochondria and improving the phosphatase and tensin homologue induced putative kinase 1 (PINK1) - Parkin mitophagy signaling pathway, while downregulating amyloid, p-tau, and autophagy marker P62 levels. Conclusion: DA4-JC is a promising drug for the treatment of AD.

Enzyme engineering toward catalytic-tetrad residues usually results in activity loss. Unexpectedly, we found that a directed evolution campaign yielded a beneficial residue A100 in KmCR (a carbonyl reductase from Kluyveromyces marxianus ZJB14056), which is a residue of catalytic tetrad and conserved according to multiple sequence alignment. Inspired by this finding, we performed saturation mutagenesis on all the four residues of catalytic tetrad of KmCR. A number of variants with improved enzymatic activities were obtained. Among them, the variant KmCR_A100S exhibited increased catalytic efficiency (kcat /KM = 47.3 s-1 ·mM-1 ), improved stereoselectivity (from moderate selectivity (deP = 66.7%) to strict (S)-selectivity (deP > 99.5%)), and extended substrate scope, compared to those of KmCR_WT. In silico analysis showed that a relay system was rebuilt in KmCR via the beneficial residue S100. Furthermore, comparison of 11 protein engineering campaigns indicated that the beneficial position is easily overlooked due to the long distance (>10 A) from ketone substrates. Since CRs share similar catalytic mechanism, the knowledge gained from this study has universal significance to CR engineering.

Purpose: Considerable variations in methylation profile have been found in various cancers to modulate tumorigenesis and affect prognosis. To provide a theoretical basis for early detection, prognosis evaluation and targeted treatment for patients with pancreatic ductal adenocarcinoma: PDAC, this study identified methylation-driven genes in PDAC and explored their prognostic performance. Methods: The methylation, expression and clinical data of PDAC patients were extracted from TCGA database. Based on the β-mixture model of the MethylMix R package, the differential methylation status and connection between methylation and expression degree were examined to screen out methylation-driven genes in PDAC. COX analyses and lasso regressions were applied to construct a linear risk model based on methylation-driven genes. Univariate and multivariate analyses were performed to ensure the risk model was an independent prognostic factor. Joint survival analyses of methylation and gene expression were conducted to explore the prognostic value of component genes. The methylation sites in the key genes were also investigated. Results: A total of 118 methylation-driven genes in PDAC were identified, and two genes (FOXI2, MYEOV) constituted the risk model whose AUC was 0.722 at one year of overall survival rate, displaying a better performance on survival prediction than other clinical features. Further survival analyses demonstrated that the expression of MYEOV and combined methylation and expression levels of the genes MYEOV and FOXI2 can be potential biomarkers for survival prediction and targets of drug manipulation of PDAC patients. Close relationships were discovered between two sites in MYEOV and one site in FOXI2 and the prognosis of PDAC patients. Conclusion: Concentrating on DNA methylation, our study identified potential biomarkers and developed a reliable short-term predictive model for prognosis of PDAC patients.

Objective To understand the characteristics of DNA methyltransferase 3a (DNMT3a) in thymoma associated Myasthenia Gravis reveal its transcriptional regulator network as while as analyze the effect of DNMT3a on Rel/ nuclear factor-kappaB family (RelA/RelB) and its downstream autoimmune regulatory factor (Aire). Methods Tissues of 30 patients with thymoma, with or without myasthenia gravis (MG), were collected and the DNMT3a protein expression were evaluated through immunohistochemistry. We performed mRNA expression profiling microarray detection and analysis, and integrated the analysis by constructing protein-protein interaction networks and the integration with other database. We identified molecular difference between low and high DNMT3a in the thymoma by heatmap. We also performed PCR validation in thymoma tissues. The DNMT3a-shRNA plasmid was transfected into TEC cells, and these cells were treated with 5-aza-2-deoxycytidine, a blocker of DNMT3a. After the down-regulation of DNMT3a in TEC cells, the transcript and protein levels of RelA, RelB, Aire, and CHRNA3 were evaluated by western blotting. In addition, changes in gene expression profiles were screened through microarray technology. We performed differential gene analysis in the thymoma cohort by heatmap with R (v.4.3.0) software. Results In 30 matched tissue specimens, the expression of DNMT3a protein in thymoma with MG was lower than that in thymoma. Through mRNA expression profiling analysis, we constructed a co-expression network of DNMT3a and found direct interaction between IKZF1 and DNMT3a, and this co-expression relationship was overlappted with Cistrome DB database. We found up-regulation of 149 mRNAs and repression of 177 mRNAs in thymoma with MG compared with thymoma. Gene ontology and pathway analysis show the involvement of a multitude of genes in the mis-regulation of MG-related pathways. RNA interference significantly reduced the level of mRNA of DNMT3a, which proved that plasmid DNMT3a was effective. In comparison to the control group, the levels of DNMT3a, Aire, and CHRNA3 mRNA and protein in TEC cells transfected with DNMT3a-shRNA interference plasmid were significantly decreased, while the expression level of RelA and RelA/RelB was significantly increased. Conclusions Our study reveals the DNMT3a-NF-κB pathway has a major effect on MG, and can be used as a marker for diagnosis as well as a target for MG treatment.

Objective This study aims to determine the mechanism of ISLR on the progression of colon cancer. Methods TCGA database was used to analyze ISLR expression in colon cancer tumor tissues. QRT-PCR and western blotting were used to detect ISLR expression in colon cancer cells. CCK-8, colony formation, EDU, wound healing and transwell assays were used to measure cell viability, proliferation, migration and invasion of colon cancer cells, respectively. The signaling pathway enrichment analysis of ISLR was analyzed on the basis of the KEGG database. The protein expression of genes related to signaling pathway was measured by western blotting. Results Results of TCGA analysis, qRT-PC and western blotting showed that ISLR was upregulated in colon cancer tumor tissues and cells. High level of ISLR was related to low overall survival of patients with colon cancer. ISLR silence significantly inhibited cell viability, proliferation, migration and invasion of colon cancer cells. ISLR overexpression markedly enhanced the cell viability, proliferation, migration and invasion of colon cancer cells. KEGG database analyzed showed that ISLR can activate the EMT signaling pathway. Inhibition of the EMT signaling pathway can suppress the growth, migration, and invasion of colon cancer cells and eliminate the promoted effect of ISLR overexpression on colon cancer progression. Conclusion ISLR promotes the progression of colon cancer by activating the EMT signaling pathway.

The mechanism of color pattern formation in animals is complicated. Oujiang color common carp (Cyprinus carpio var. color) is an excellent fish model for studying the molecular mechanism of pigmentation. Solute carrier family 24 member 5 (SLC24A5) was identified as a key gene in regulating melanophore development and melanin distribution in organisms. In this study, functional experiments were conducted on the SLC24A5 gene in Oujiang color common carp. The full-length of SLC24A5 mRNA was cloned and obtained. In Oujiang color common carp, qRT-PCR and whole-mount in situ hybridization indicated that SLC24A5 was highly expressed in tissues in the presence of melanin and initiated its expression along with the presence of melanin during embryo development. The SLC24A5 knocked-out via CRISPR/cas9 caused an obvious loss of melanophore in larva and adult zebrafish. In addition, the homozygous SLC24A5−/− zebrafish mutant phenotype was rescued by the over expression of the SLC24A5 from Oujiang color common carp. Results suggested that SLC24A5 is essential for regulating melanophore development and that the functional and structures of SLC24A5 are conserved in Cyprinidae fish.

Circular RNA is a newly discovered member of non‐coding RNA (ncRNA) and regulates the target gene by acting as a micro‐RNA sponge. It plays vital roles in various diseases. However, the functions of circular RNA in non‐small cell lung cancer (NSCLC) remain still unclear. Our data showed that circ‐WHSC1 was highly expressed in NSCLC cells and tissues. Both in vitro and in vivo experiments showed that circ‐WHSC1 promoted NSCLC proliferation. circ‐WHSC1 also promoted the migration and invasion of lung cancer cells. Through bioinformatic analysis and functional experiments, we showed that circ‐WHSC1 could act as a sponge for micro‐RNA‐7 (miR‐7) and regulate the expression of TAB2 (TGF‐beta activated kinase one binding protein two). Inhibition of the circ‐WHSC1/miR‐7/TAB2 pathway could effectively attenuate lung cancer progression. In summary, this study confirmed the existence and oncogenic function of circ‐WHSC1 in NSCLC. The research suggests that the circ‐WHSC1/miR‐7/TAB2 axis might be a potential target for NSCLC therapy.

Gut microbial dysbiosis has been linked to many noncommunicable diseases. However, little is known about specific gut microbiota composition and its correlated metabolites associated with molecular signatures underlying host response to infection. Here, we describe the construction of a proteomic risk score based on 20 blood proteomic biomarkers, which have recently been identified as molecular signatures predicting the progression of the COVID-19. We demonstrate that in our cohort of 990 healthy individuals without infection, this proteomic risk score is positively associated with proinflammatory cytokines mainly among older, but not younger, individuals. We further discover that a core set of gut microbiota can accurately predict the above proteomic biomarkers among 301 individuals using a machine learning model and that these gut microbiota features are highly correlated with proinflammatory cytokines in another independent set of 366 individuals. Fecal metabolomics analysis suggests potential amino acid-related pathways linking gut microbiota to host metabolism and inflammation. Overall, our multi-omics analyses suggest that gut microbiota composition and function are closely related to inflammation and molecular signatures of host response to infection among healthy individuals. These results may provide novel insights into the cross-talk between gut microbiota and host immune system.

The significant function of circRNAs in cancer was recognized in recent work, so a well-organized resource is required for characterizing the interactions between circRNAs and other functional molecules (such as microRNA and RNA-binding protein) in cancer. We previously developed cancer-specific circRNA database (CSCD), a comprehensive database for cancer-specific circRNAs, which is widely used in circRNA research. Here, we updated CSCD to CSCD2 (http://geneyun.net/CSCD2 or http://gb.whu.edu.cn/CSCD2), which includes significantly more cancer-specific circRNAs identified from a large number of human cancer and normal tissues/cell lines. CSCD2 contains >1000 samples (825 tissues and 288 cell lines) and identifies a large number of circRNAs: 1 013 461 cancer-specific circRNAs, 1 533 704 circRNAs from only normal samples and 354 422 circRNAs from both cancer and normal samples. In addition, CSCD2 predicts potential miRNA–circRNA and RBP–circRNA interactions using binding motifs from >200 RBPs and 2000 microRNAs. Furthermore, the potential full-length and open reading frame sequence of these circRNAs were also predicted. Collectively, CSCD2 provides a significantly enhanced resource for exploring the function and regulation of circRNAs in cancer.

To clarify the mechanism of semicarbazide-modified α-lactalbumin (SEM-LA)-mediated cytotoxicity, we investigated its effect on human U937 leukemia cells and MCF-7 breast cancer cells in the current study. SEM-LA induced apoptosis in U937 cells, which showed increased NOX4 expression, procaspase-8 degradation, and t-Bid production. FADD depletion inhibited SEM-LA-elicited caspase-8 activation, t-Bid production, and cell death, indicating that SEM-LA activated death receptor-mediated apoptosis in U937 cells. SEM-LA stimulated Ca2+-mediated Akt activation, which in turn increased Sp1- and p300-mediated NOX4 transcription. The upregulation of NOX4 expression promoted ROS-mediated p38 MAPK phosphorylation, leading to protein phosphatase 2A (PP2A)-regulated tristetraprolin (TTP) degradation. Remarkably, TTP downregulation increased the stability of TNF-α mRNA, resulting in the upregulation of TNF-α protein expression. Abolishment of Ca2+-NOX4-ROS axis-mediated p38 MAPK activation attenuated SEM-LA-induced TNF-α upregulation and protected U937 cells from SEM-LA-mediated cytotoxicity. The restoration of TTP expression alleviated the effect of TNF-α upregulation and cell death induced by SEM-LA. Altogether, the data in this study demonstrate that SEM-LA activates TNF-α-mediated apoptosis in U937 cells through the NOX4/p38 MAPK/PP2A axis. We think that a similar pathway can also explain the death of MCF-7 human breast cancer cells after SEM-LA treatment.

Beijing will host the 2022 Winter Olympics, and China strengthens research on various aspects to allow their athletes to compete successfully in winter sport. Simultaneously, Government-directed initiatives aim to increase public participation in recreational winter sport. These parallel developments allow research to advance knowledge and understanding of the physiological determinants of performance and health related to winter sport. Winter sport athletes often conduct a substantial amount of training with high volumes of low-to-moderate exercise intensity and lower volumes of high-intensity work. Moreover, much of the training occur at low ambient temperatures and winter sport athletes have high risk of developing asthma or asthma-related conditions, such as exercise-induced bronchoconstriction. The high training volumes require optimal nutrition with increased energy and dietary protein requirement to stimulate muscle protein synthesis response in the post-exercise period. Whether higher protein intake is required in the cold should be investigated. Cross-country skiing is performed mostly in Northern hemisphere with a strong cultural heritage and sporting tradition. It is expected that innovative initiatives on recruitment and training during the next few years will target to enhance performance of Chinese athletes in classical endurance-based winter sport. The innovation potential coupled with resourcing and population may be substantial with the potential for China to become a significant winter sport nation. This paper discusses the physiological aspects of endurance training and performance in winter sport highlighting areas where innovation may advance in athletic performance in cold environments. In addition, to ensure sustainable development of snow sport, a quality ski patrol and rescue system is recommended for the safety of increasing mass participation.

BACKGROUND: Blueberry is among the fastest growing fruit crops in the world, which is beneficial to human health and attracts extensive interests. In contrast to its rapid development and utilization, availability of molecular and genetic resources for blueberries are still scarce. OBJECTIVE: In present report, transcriptomic profiling of four widely cultivated varieties of Rabbiteye and Southern Highbush blueberries were characterized to assist the breeding programs. METHODS: Both de novo and reference-based assembly were conducted to generate the genetic resources that can be used in the further functional and breeding studies. RESULTS: De novo and reference-based assembly found average 136,350 and 158,123 non-redundant transcripts, respectively. Average 57,668 de novo assembled transcripts can be functionally annotated by homology search with different databases. We further detected 6,268 polymorphic simple sequence repeats, 566,913 high-quality single nucleotide polymorphisms and 88,662 insertion and deletions among the four varieties with comparison to a recently released reference genome of blueberry. Differentially expressed genes analysis showed that varieties of same species show less differences within species but larger differences between species. CONCLUSIONS: These comprehensive and high-quality genetic resources will contribute to a wide range of genetics and molecular breeding studies in blueberries.