LA-D-B1,a novel Abemaciclib derivative,exerts anti-breast cancer effects through CDK4/6

Background:Regulatory proteins involved in human cellular division and proliferation,cyclin-dependent kinases 4 and 6(CDK4/6)are overexpressed in numerous cancers,including triple-negative breast cancer(TNBC).TNBC is a common pathological subtype of breast cancer that is prone to recurrence and metastasis,and has a single treatment method.As one of the CDK4/6 inhibitors,abemaciclib can effectively inhibit the growth of breast tumors.In this study,we synthesized LA-D-B1,a derivative of Abemaciclib,and investigated its anti-tumor effects in breast cancer.Methods:Cellular viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay.Cell cloning and migration abilities were determined by colony formation assay and wound healing assay.Cell invasion abilities and adhesion were determined by cell invasion assay and cell adhesion assay.The impact of compound LA-D-B1 on cell proliferation and the cell cycle was analyzed through Western blotting,which quantified the levels of proteins associated with the cyclin-dependent kinase(CDK)4/6-cyclin D-Rb-E2F pathway.The in vivo anti-tumor activity of compound LA-D-B1 was investigated using a chick chorioallantoic membrane(CAM)model.Results:The study demonstrated that LA-D-B1 effectively suppressed breast cancer cell proliferation,induced apoptosis,and caused cell cycle arrest.Furthermore,LA-D-B1 reduced the expression of key proteins in the CDK4/6-cyclin D-Rb-E2F pathway,including CDK4,CDK6 and E2F1.The results also indicated significant antitumor activity of LA-D-B1 in a transplanted tumor model.Conclusion:In this study,LA-D-B1 demonstrated a potent anti-tumor effect by effectively suppressing cell proliferation and inhibiting cell cycle progression in breast cancer.These findings highlight the potential of LA-D-B1 as a valuable compound for enhancing therapeutic outcomes and controlling the progression of breast cancer.

Malvidin Mitigates Sepsis-induced Cardiac Injury by Modulating the TLR4-iNOS-COX-2 Inflammatory Pathway and the Bax/Bcl-2/Cyto-C Mitochondrial Apoptosis Pathway in a p38 MAPK-dependent Manner

In critical care medicine,sepsis is a dangerous systemic condition that is highly prevalent and is associated with high morbidity and mortality rates^([1]).The high mortality rate associated with sepsis is closely related to multi-organ dysfunction,with heart injury being particularly critical and considered the starting point of multi-organ injury^([2]).

Insights into microbiota community dynamics and flavor development mechanism during golden pomfret(Trachinotus ovatus)fermentation based on single-molecule real-time sequencing and molecular networking analysis

Popular fermented golden pomfret(Trachinotus ovatus)is prepared via spontaneous fermentation;however,the mechanisms underlying the regulation of its flavor development remain unclear.This study shows the roles of the complex microbiota and the dynamic changes in microbial community and flavor compounds during fish fermentation.Single-molecule real-time sequencing and molecular networking analysis revealed the correlations among different microbial genera and the relationships between microbial taxa and volatile compounds.Mechanisms underlying flavor development were also elucidated via KEGG based functional annotations.Clostridium,Shewanella,and Staphylococcus were the dominant microbial genera.Forty-nine volatile compounds were detected in the fermented fish samples,with thirteen identified as characteristic volatile compounds(ROAV>1).Volatile profiles resulted from the interactions among the microorganisms and derived enzymes,with the main metabolic pathways being amino acid biosynthesis/metabolism,carbon metabolism,and glycolysis/gluconeogenesis.This study demonstrated the approaches for distinguishing key microbiota associated with volatile compounds and monitoring the industrial production of high-quality fermented fish products.

Analysis of quality-related proteins in golden pompano(Trachinotus ovatus)fillets with modified atmosphere packaging under superchilling storage

Here,we aimed to study the changes in proteome of golden pompano fillets during post-mortem storage.Tandem mass tags(TMT)-labeled quantitative proteomic strategy was applied to investigate the relationships between protein changes and quality characteristics of modified atmosphere packaging(MAP)fillets during superchilling(-3°C)storage.Scanning electron microscopy was used to show that the muscle histology microstructure of fillets was damaged to varying degrees,and low-field nuclear magnetic resonance was used to find that the immobilized water and free water in the muscle of fillets changed significantly.Total sulfhydryl content,TCA-soluble peptides and Ca2+-ATPase activity also showed that the fillet protein had a deterioration by oxidation and denaturation.The Fresh(FS),MAP,and air packaging(AP)groups were set.Total of 150 proteins were identified as differential abundant proteins(DAPs)in MAP/FS,while 209 DAPs were in AP/FS group.The KEGG pathway analysis indicated that most DAPs were involved in binding proteins and protein turnover.Correlation analysis found that 52 DAPs were correlated with quality traits.Among them,8 highly correlated DAPs are expected to be used as potential quality markers for protein oxidation and water-holding capacity.These results provide a further understanding of the muscle deterioration mechanism of packaging golden pompano fillets during superchilling.

Identifying and validating MMP family members(MMP2,MMP9,MMP12,and MMP16)as therapeutic targets and biomarkers in kidney renal clear cell carcinoma(KIRC)

Kidney Renal Clear Cell Carcinoma(KIRC)is a malignant tumor that carries a substantial risk of morbidity and mortality.The MMP family assumes a crucial role in tumor invasion and metastasis.This study aimed to uncover the mechanistic relevance of the MMP gene family as a therapeutic target and diagnostic biomarker in Kidney Renal Clear Cell Carcinoma(KIRC)through a comprehensive approach encompassing both computational and molecular analyses.STRING,Cytoscape,UALCAN,GEPIA,OncoDB,HPA,cBioPortal,GSEA,TIMER,ENCORI,DrugBank,targeted bisulfite sequencing(bisulfite-seq),conventional PCR,Sanger sequencing,and RT-qPCR based analyses were used in the present study to analyze MMP gene family members to accurately determine a few hub genes that can be utilized as both therapeutic targets and diagnostic biomarkers for KIRC.By performing STRING and Cytohubba analyses of the 24 MMP gene family members,MMP2(matrix metallopeptidase 2),MMP9(matrix metallopeptidase 9),MMP12(matrix metallopeptidase 12),and MMP16(matrix metallopeptidase 16)genes were denoted as hub genes having highest degree scores.After analyzing MMP2,MMP9,MMP12,and MMP16 via various TCGA databases and RT-qPCR technique across clinical samples and KIRC cell lines,interestingly,all these hub genes were found significantly overexpressed at mRNA and protein levels in KIRC samples relative to controls.The notable effect of the up-regulated MMP2,MMP9,MMP12,and MMP16 was also documented on the overall survival(OS)of the KIRC patients.Moreover,targeted bisulfite-sequencing(bisulfite-seq)analysis revealed that promoter hypomethylation pattern was associated with up-regulation of hub genes(MMP2,MMP9,MMP12,and MMP16).In addition to this,hub genes were involved in various diverse oncogenic pathways.The MMP gene family members(MMP2,MMP9,MMP12,and MMP16)may serve as therapeutic targets and prognostic biomarkers in KIRC.

A Fluorescent Probe based on a Carbazole Derivative for Detecting Brilliant Blue in Food

Here a fluorescent probe based on a carbazole derivative(CNS)was developed to increase the detection range and reduce the detection limit of brilliant blue.Characteristics of CNS are studied.Due to the quenching ability of colorants,CNS shows an excellent current response to brilliant blue(from 1 to 10μM)with a detection limit of 2.7×10^(-8)mol/L(3σ/k)in the conditions of a 1:1 volume ratio of water to tetrahydrofuran.And the stability and reproducibility of CNS in the detection of actual samples indicate great potential for application.

Sources and transformations of nitrite in the Amundsen Sea in summer 2019 and 2020 as revealed by nitrogen and oxygen isotopes

In this study,the nitrogen and oxygen isotope compositions of nitrite in the upper 150 m water column of the Amundsen Sea in the summer of 2019 and 2020 were measured to reveal the distribution and transformation of nitrite in the euphotic zone of the Southern Ocean.We found that primary nitrite maxima(PNMs)are widely present in the Amundsen Sea,where the depth of occurrence deepens from east to west and nitrite concentrations increases.Evidence from dual isotopes suggests that the formation of PNMs in all regions of the Amundsen Sea is dominated by ammonia oxidation.More importantly,the nitrogen and oxygen isotope compositions of nitrite in the Amundsen Sea mixed layer are abnormal,and their depth profiles are mirror symmetrical.Isotopic anomalies exhibit spatial variations,with central surface water having the lowest nitrogen isotope composition(−89.9‰±0.2‰)and western surface water having the highest oxygen isotope composition(63.3‰±0.3‰).Isotopic exchange reaction between nitrate and nitrite is responsible for these isotope anomalies,as both nitrogen and oxygen isotopes have large isotopic fractionation and opposite enrichment effects.This proves that isotopic exchange reaction operates extensively in different regions of the Amundsen Sea.Our study highlights the unique role of dual isotopes of nitrite in deepening the understanding of nitrogen cycle.Further studies on ammonia oxidation and isotopic exchange between nitrate and nitrite are warranted in the future to understand their roles in the nitrogen cycle in the Southern Ocean.

Achieving Physical Layer Security Against Location Unknown Eavesdroppers via Friendly Jammer

Dear Editor, This letter is concerning friendly jamming unmanned aerial vehicles(UAVs) to assist in the safe communication of UAV base stations. Due to the openness of UAV wireless communication, it is vulnerable to attacks leading to information disclosure or blockage. To address this issue, friendly jamming UAVs can assist UAV base stations and improve the security of wireless communications.

连续排水边界下成层地基一维非线性固结解析解

为了研究边界排水时间效应对固结过程的影响,引入连续排水边界研究成层地基一维非线性固结问题。通过引入孔隙比与有效应力之间的非线性关系,建立了瞬时荷载作用下成层地基固结问题的控制方程,进而获得按沉降定义和按超静孔隙水压力定义的平均固结度以及成层地基沉降的解析解。通过与有限差分解析解对比以及现有解析解对比验证了所得解析解的正确性。基于所得解,通过详细的参数分析研究了成层地基一维非线性固结特性。结果表明,排水界面参数对成层地基的固结性状有很大影响,固结速率随界面参数的增大而增大。在连续排水边界条件下,按沉降定义的平均固结度随有效应力比的增大而增大,而按超静孔隙水压力定义的平均固结度随有效应力比的增大而减小。

Height connection across sea by using satellite altimetry data sets,ellipsoidal heights,astrogeodetic deflections of the vertical,and an Earth Gravity Model

Islands and the mainland are separated by seas,and the distances between them might be so long that the height on the mainland cannot be exactly translated to the islands,resulting in different height systems on the mainland and the islands.In this study,we used astrogeodetic deflections of the vertical and ellipsoidal heights of points on the mainland and island near their coastlines to implement height connection across sea areas.First,the modeled gravity and modeled astrogeodetic vertical deflections of segmentation points along connecting routes over the sea between the mainland and the island were determined by Earth Gravity Model(EGM),and the ellipsoidal heights of segmentation points were determined by the satellite altimetry data sets.Second,we used a linear interpolation model to increase the precision of the vertical deflections of segmentation points.Third,we computed the geopotential difference of points between the mainland and the island using a method derived from geopotential theory and the astronomical leveling principle.Finally,we estimated the normal height of the point on the island using the geopotential-difference iterative computation approach.Using observed data of normal heights,ellipsoidal heights,and astrogeodetic vertical deflections referring to height sites in Qingdao,Shandong Province,we conducted a numerical experiment involving the normal height connection across sea regions.We determined the data of the ellipsoidal heights and gravity of segmentation points along the connecting route across the water in the numerical experiment using DTU10.The distance of the height connection across the sea was approximately 10.5 km.According to China's official leveling specifications,the experimental results met the criterion of third-class leveling precision.

Novel insight into the formation and inhibition mechanism of dipeptidyl peptidase-Ⅳ inhibitory peptides from fermented mandarin fish(Chouguiyu)

Fermented foods are a potential source to produce novel dipeptidyl peptidase-IV inhibitory peptides(D4IPs).In this study,the fermented mandarin fish(Chouguiyu)was used to screen D4IPs and their formation mechanism was studied by metagenomics and peptidomics.A total of 400 D4IPs with DPP-IV inhibition structure and high hydrophobicity were identified.The correlation network map showed that Lactococcus,Bacillus,Lysobacter,Pelagivirga,Kocuria,Escherichia,Streptococcus,and Peptostreptococcus were significantly correlated with the most D4IPs.Four stable D4IPs,including KAGARALTDAETAT,GEKVDFDDIQK,VVDADEMYLKGK,and GQKDSYVGDEAQ were respectively from the precursor proteins parvalbumin,troponin,myosin,and actin,and were mainly formed by the hydrolysis of subtilisin(EC 3.4.21.62),aspartic proteinase(EC 3.4.23.1),thermolysin(EC 3.4.24.27),oligopeptidase B(EC 3.4.21.83),and proteinase P1(EC 3.4.21.96)from Bacillus,Kocuria,Lysobacter,Lactococcus,and Peptostreptococcus.The inhibition mainly resulted from the hydrogen bond and salt bridge between D4IPs and DPP-IV enzyme.This study provides important information on the proteases and related microbial strains to directionally prepare D4IPs in Chouguiyu.

Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis in Manila Clam Ruditapes philippinarum Under Hypoxic Stress

Quantitative real-time PCR(qRT-PCR)has been widely used for gene expression analysis,and selection of reference genes is a key point to obtain accurate results.To find out optimal reference genes for qRT-PCR in Manila clam Ruditapes philippinarum in response to hypoxia,different tissues were used and compared to evaluate the stability of candidate reference genes under low oxygen stress(DO 0.5mgL^(−1) and DO 2.0mgL^(−1))and normal condition(DO 7.5mgL^(−1)).Seven candidate reference genes were selected to evaluate the stability of their expression levels.The reference genes were evaluated by Delta Ct,BestKeeper,NormFinder and geNorm,and then screened by RefFinder calculation.Under hypoxic stress of 0.5mgL^(−1),the most suitable reference gene for gill and hepatopancreas was RPL31,and the optimal reference genes for axe foot and adductor muscle were TUB and HIS,respectively.For hypoxic stress of 2.0mgL^(−1),the most stable reference gene for gill and hepatopancreas was RPL31,and the optimal reference genes for axe foot and adductor muscle were RPS23 and EF1A,respectively.At the normal condition,HIS and EF1A were identified as the optimal internal reference genes in gill and hepatopancreas respectively,and GFRP2 was the best internal reference gene for axe foot and adductor muscle.The present findings will provide important basis for the selection of reference genes for qRT-PCR analysis of gene expression level in bivalves under hypoxic stress,which might be helpful for the analysis of other molluscs too.

A Rapid and Accurate CRISPR/Cas12b-Mediated Genotyping Assay for the Methylenetetrahydrofolate Reductase Gene Polymorphism C677T

Human methylenetetrahydrofolate reductase(MTHFR),an essential enzyme in folate metabolism,catalyzes the conversion of5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate.Several polymorphisms in the MTHFR gene have been found to be associated with reduced enzyme activity and related to folic acid intake in relevant populations[1].

The Response of Bio-Component Production of Nannochloris oculata to the Combinations of Monochromatic Light

Illuminating conditions are crucial factors affecting the microalgal growth and biosynthesis.The effects of illuminating spectral quality on the growth and bio-component production of Nannochloris oculata were investigated.The results indicated that a high proportion of Red-light enhanced the pigments and carbohydrate production but reduced those of the biomass and lipid.Mono-chromatic Blue-light has advantageous effects on lipid production compared with Red-and White-light.The optimal light spectrum for the protein production was the combination of Red-and Blue-light at a ratio of 4:3 or 5:2.Among the seven fatty acids identified from N.oculata,the contents of C16:0,C18:0,and C18:3(n-3)in the lipid were inhibited by the increased proportion of Red-light while those of C18:2(n-9),C16:2(n-6),and C20:0 were inhibited by Blue-light.Monochromatic Red-and Blue-light and their com-binations were proposed as a promising illuminating strategy for the large-scale cultivation aiming for various bio-components.

Free-Fall Penetration Behaviors of A New Dynamically Installed Plate Anchor in Marine Clay

A new dynamically installed plate anchor, the Flying Wing Anchor~?, has been developed as a sustainable anchor concept for deep-water offshore wind turbines. The anchor is firstly installed by free-fall penetration and then followed by drag embedment. If the anchor is subjected to environmental loads, it dives deeper to mobilize a higher capacity. This study presents a series of free-fall penetration tests with model anchors in different weights to assess the anchor behavior during the free-fall penetration performance in one-layer soil with a constant shear strength profile. Anchor velocities and embedment depths were measured by a magnetometer. An energy-based model and a force-based model were calibrated against the test results of model anchors with different weights. Based on the calibrated force-based model, a series of design charts were developed to estimate the embedment depth of anchors in different sizes and with different impact velocities in various marine clays. The framework to plot design charts presented herein can be potentially applied to other dynamically installed anchors to predict embedment depth in engineering practice.

Phytoplankton community dynamics during Alexandrium blooms in 2019 off the Qinhuangdao coast,Bohai Sea,China

Alexandrium blooms in the northwest area of the Bohai Sea(Qinhuangdao coastal area),China,produce large amounts of toxins that could be enriched in shellfish and consequently harm human bodies.To understand the succession of the phytoplankton community structure during Alexandrium bloom events in the northwest area of the Bohai Sea off Qinhuangdao from April 2 to May 7,2019,microscopy observations and high-performance chromatography(HPLC)-pigment analysis were performed.Sixty species of phytoplankton were identified,mainly diatoms and dinofl agellates.The abundance of Alexandrium reached the maximum on April 16(3.3×10^(3) cells/L).HPLC-pigment CHEMTAX analysis showed that the phytoplankton community was composed mainly of diatoms,dinofl agellates,prasinophytes,and cryptophytes.Diatoms were the main contributor to the total Chl-a pool.There was a downward trend for the proportion of diatom biomass to the total Chl-a pool,followed by an upward trend.The proportion of dinoflagellate biomass showed the opposite trend,whereas that of the prasinophyte biomass presented an obvious increasing trend.Temperature,nutrients,and nutrient structures were the main factors on the distribution of different phytoplankton groups in the study area as shown in the redundancy analysis.This work illustrates the succession of phytoplankton community structures during Alexandrium blooms and provided a theoretical basis for studies on the mechanism underlying the outbreak of harmful algal blooms in sea areas.

Duplex Detection of Vibrio Cholerae and Vibrio Parahaemolyticus by Real-time Recombinase Polymerase Amplification

Vibrio cholerae and Vibrio parahaemolyticus are the two most commonly reported pathogens in seafood[1]. Consuming raw or undercooked seafood contaminated with these two Vibrio species can cause food poisoning, posing the risk of severe gastrointestinal illness and death[2-3]. Therefore,precise and reliable methods for detecting V. cholerae and V. parahaemolyticus contamination in seafood are essential for controlling food safety.

In vitro Antioxidant Effects of Porphyra haitanensis Peptides on H_2O_2-Induced Damage in HepG2 Cells

In this study,protein from Porphyra haitanensis was used as raw material to prepare an antioxidant peptide,and its antioxidant activity was evaluated in vitro.A model of H_2O_2-induced oxidative damage in Hep G2 cells was established,and the effects of Porphyra haitanensis hydrolysates (PHHs) on superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were detected.Finally,the structure of PHHs was identified by ESI-MS/MS.The results showed that the 1,1-diphenyl-2-pyridylhydrazine(DPPH)-free radical-scavenging ability of PHHs was the strongest (59.28%at 1.0 mg m L~(-1)) when hydrolyzed with an acidic protease for 4 h.PHHs with different concentrations had protective effects on H_2O_2-induced damage to Hep G2 cells,and the protective effect was enhanced with increasing concentrations.When the level was 400μg m L~(-1),the cell survival rate was as high as 88.62%.Moreover,PHHs can significantly reduce oxidative damage to Hep G2 cells by H_2O_2,improve SOD activity,and reduce MDA content.The tetrapeptide Asp-Lys-Ser-Thr,with a molecular weight of 448 Da,was identified as an important fraction of PHHs by high-resolution mass spectrometry.

Effects of Dietary Protein and Lipid Levels on Growth Performance, Muscle Composition, Immunity Index and Biochemical Index of the Greenfin Horse-Faced Filefish (Thamnaconus septentrionalis) Juvenile

The effects of different dietary protein and lipid levels on juvenile greenfin horse-faced filefish(Thamnaconus septentrionalis)was experimentally evaluated in a 56-day study.Fish(initial weight 6.47±0.56 g)were fed with nine different diet combinations containing three varying levels of protein(37%,43%,49%)and lipids(4%,7%,10%)in triplicates,where the protein-to-energy(P/E)ratios of the diets varied in the range of 19.17-26.99mgkJ−1.Results showed that fish fed diets containing 49%protein and 10%lipid exhibited significantly lower feed intake than those fed other diets.The weight gain of fish fed diets with 43%protein was significantly higher than that of fish fed 37%or 49%protein diets.While no significant change in weight gain was observed when lipid content was increased from 4%to 7%,weight gain decreased significantly when lipid increased from 7%to 10%.The protein efficiency ratio and feed efficiency were highest when the fish were fed the diet with 49%protein and 7%lipids.Muscle crude lipid content was positively correlated with dietary protein level regardless of lipid level.The diet of 43%protein or 7%lipids significantly reduced the activity of alanine aminotransferase and aspartate aminotransferase in liver.The contents of aspartate aminotransferase,superoxide dismutase and malondialdehyde were significantly increased from 4%to 10%lipids in plasma.The plasma total cholesterol and triglyceride contents were positively correlated with dietary lipid level at the same protein level.Our comprehensive analysis concluded that a diet with 43%protein and 7%lipid,with a P/E ratio of 23.07mgkJ−1,significantly promote the growth performance,feed utilization and health of juvenile greenfin horse-faced filefish.

Newly recorded bloom-forming dinoflagellate Gymnodinium impudicum in Haizhou Bay,Yellow Sea,China

With the development of industrialization and aquaculture in Jiangsu and Shandong Provinces along the South Yellow Sea coast,China,eutrophication has greatly intensified in the region,resulting in frequent occurrence of diverse harmful algal blooms.An algal bloom formed by a chain-forming dinofl agellate species was recorded in the Haizhou Bay,South Yellow Sea,in September 2020.The causative species was isolated and studied in morphology,molecular phylogeny,pigment profile,presence of paralytic shellfish toxins,and acute toxicity.The loop-shaped apical groove running anticlockwise around the apex,the presence of peridinin as characteristic pigment,as well as a single phylogenic clade of 28S ribosomal DNA(100%posterior probability),defined this species as Gymnodinium impudicum,a non-toxic species that exhibited no obvious biotoxicity to the rotifer Brachionus plicatilis,the copepod Artemia salina,and the shrimp Neomysis awatschensis.Gymnodinium impudicum is typically distributed in coastal waters with high nitrate concentrations,where it reaches a maximum density of 2.6×10~5 cells/L.This is the first report of a G.impudicum bloom in the Yellow Sea;however,G.impudicum blooms may have been misidentified or underreported in Haizhou Bay due to the species morphological similarity with G.catenatum.A combination of multiple methods is recommended to accurately identify new algal bloom species.