An efficient method for constructing a random insertional mutant library for forward genetics in Nannochloropsis oceanica

Insertional mutation,phenotypic evaluation,and mutated gene cloning are widely used to clone genes from scratch.Exogenous genes can be integrated into the genome during non-homologous end joining(NHEJ)of the double-strand breaks of DNA,causing insertional mutation.The random insertional mutant library constructed using this method has become a method of forward genetics for gene cloning.However,the establishment of a random insertional mutant library requires a high transformation efficiency of exogenous genes.Many microalgal species show a low transformation efficiency,making constructing random insertional mutant libraries difficult.In this study,we established a highly efficient transformation method for constructing a random insertional mutant library of Nannochloropsis oceanica,and tentatively tried to isolate its genes to prove the feasibility of the method.A gene that may control the growth rate and cell size was identified.This method will facilitate the genetic studies of N.oceanica,which should also be a reference for other microalgal species.

The inhibitory effect of adenosine on tumor adaptive immunity and intervention strategies

Adenosine(Ado)is significantly elevated in the tumor microenvironment(TME)compared to normal tissues.It binds to adenosine receptors(AdoRs),suppressing tumor antigen presentation and immune cell activation,thereby inhibiting tumor adaptive immunity.Ado downregulates major histocompatibility complex II(MHC II)and co-stimulatory factors on dendritic cells(DCs)and macrophages,inhibiting antigen presentation.It suppresses anti-tumor cytokine secretion and T cell activation by disrupting T cell receptor(TCR)binding and signal transduction.Ado also inhibits chemokine secretion and KCa3.1 channel activity,impeding effector T cell trafficking and infiltration into the tumor site.Furthermore,Ado diminishes T cell cytotoxicity against tumor cells by promoting immune-suppressive cytokine secretion,upregulating immune checkpoint proteins,and enhancing immune-suppressive cell activity.Reducing Ado production in the TME can significantly enhance anti-tumor immune responses and improve the efficacy of other immunotherapies.Preclinical and clinical development of inhibitors targeting Ado generation or AdoRs is underway.Therefore,this article will summarize and analyze the inhibitory effects and molecular mechanisms of Ado on tumor adaptive immunity,as well as provide an overview of the latest advancements in targeting Ado pathways in anti-tumor immune responses.

Author correction to‘Mevalonate improves anti-PD-1/PD-L1 efficacy by stabilizing CD274 mRNA'[Acta Pharmaceutica Sinica B 13(2023)2585-2600]

The authors regret that one author was missed in the author list when preparing the manuscript,while the foundation to him was mentioned in the section of Acknowledgments.Dr.Qinjie Weng should be added to the author list because of his contribution in animal study and his foundation help us to complete this study.The authors sincerely apologize for any inconvenience caused to the journal and readers.

Interaction between neutrophil extracellular traps and cardiomyocytes contributes to atrial fibrillation progression

Atrial fibrillation(AF)is a frequent arrhythmia associated with cardiovascular morbidity and mortality.Neutrophil extracellular traps(NETs)are DNA fragments with cytoplasm proteins released from neutrophils,which are involved in various cardiovascular diseases.To elucidate the role of NETs in AF,we investigated the effect of NETs on AF progression and the secretion of NETs in AF.Results showed that:NETs induced the autophagic apoptosis of cardiomyocytes,and NETs also led to mitochondrial injury by promoting mitochondrial depolarization and ROS production.Ongoing tachy-pacing led to the structural loss of cardiomyocytes and provided potent stimuli to induce NETs secretion from neutrophils.In the meanwhile,increased Ang II in AF facilitated NETs formation through the upregulation of AKT phosphorylation,while it could not directly initiate NETosis as the autophagy was not induced.In vivo,DNase I was administrated to abrogate NETs formation,and AF-related fibrosis was ameliorated as expected.Correspondingly,the duration of the induced AF was reduced.Our study addresses the formation mechanism of NETs in AF and demonstrates the lethal effects of NETs on cardiomyocytes through the induction of mitochondrial injury and autophagic cell death,which comprehensively describes the positive feedback comprised of NETs and stimuli secreted by cardiomyocytes that sustains the progression of AF and AF related fibrosis.

Mevalonate improves anti-PD-1/PD-L1 efficacy by stabilizing CD274 mRNA

Mevalonate metabolism plays an important role in regulating tumor growth and progression;however,its role in immune evasion and immune checkpoint modulation remains unclear.Here,we found that non-small cell lung cancer(NSCLC)patients with higher plasma mevalonate response better to antiPD-(L)1 therapy,as indicated by prolonged progression-free survival and overall survival.Plasma mevalonate levels were positively correlated with programmed death ligand-1(PD-L1)expression in tumor tissues.In NSCLC cell lines and patient-derived cells,supplementation of mevalonate significantly upregulated the expression of PD-L1,whereas deprivation of mevalonate reduced PD-L1 expression.Mevalonate increased CD274 mRNA level but did not affect CD274 transcription.Further,we confirmed that mevalonate improved CD274 mRNA stability.Mevalonate promoted the affinity of the AU-rich elementbinding protein HuR to the 3'-UTR regions of CD274 mRNA and thereby stabilized CD274 mRNA.By in vivo study,we further confirmed that mevalonate addition enhanced the anti-tumor effect of anti-PD-L1,increased the infiltration of CD8^(+)T cells,and improved cytotoxic function of T cells.Collectively,our findings discovered plasma mevalonate levels positively correlated with the therapeutic efficacy of anti-PD-(L)1 antibody,and provided the evidence that mevalonate supplementation could be an immunosensitizer in NSCLC.

Rapid fabrication of large-area and uniform surface-enhanced Raman spectroscopy substrate of Au nano-hemisphere array and its application in the detection of Malachite Green in tilapia

Although surface-enhanced Raman spectroscopy(SERS)substrates have achieved high sensitivity,it is still difficult to apply these SERS protocols to the on-site detection of real samples due to the SERS substrate being fabrication-complexed,unstable,reproducible,or unable to be applied in batch production.Here,a large-scale ordered two-dimensional array of Au nano-hemispheres was developed through electron beam vaporization of Au onto the easy-available commercialized anodic aluminum oxide(AAO)template with two-layer nanostructures of different diameters.The uniform Au nano-hemisphere is reliable for SERS detection of malachite green(MG)due to the relative standard deviation(RSD)of the SERS intensities at different locations less than 10%.With the optimized excitation wavelength,solvent effect and pH environment,the linear range of MG detection spans from 10^(-10) to 10^(-6) mol/L with a limit of detection(LOD)of 4×10^(-10) mol/L.The enhancement factor can reach 1.2×10^(6).After extraction with acetonitrile and dichloromethane,MG in the spiked tilapia was detected with satisfactory recovery.The results indicate that the Au nano-hemisphere array can be expected to greatly facilitate SERS practical applications in detecting harmful food additives and chemicals due to the advantages of chemical inertness,physical robustness,simple fabrication,controllability,large-area uniformity,and large-batch production.

Histological analysis of hypoglycemic agents on liver fibrosis in patients with non-alcoholic fatty liver disease:a systematic review

To the Editor:Non-alcoholic fatty liver disease(NAFLD)includes a spectrum of diseases:simple steatosis,nonalcoholic steatohepatitis(NASH),and the related fibrosis and cirrhosis.[1]The treatment of NAFLD should aim to reverse or prevent liver fibrosis,which is associated with liver-specific morbidity or mortality,rather than just to reduce circulating liver enzyme activities and improve NASH.

Epigenetic strategies synergize with PD-L1/PD-1 targeted cancer immunotherapies to enhance antitumor responses

Immunotherapy strategies targeting the programmed cell death ligand 1(PD-L1)/programmed cell death 1(PD-1)pathway in clinical treatments have achieved remarkable success in treating multiple types of cancer.However,owing to the heterogeneity of tumors and individual immune systems,PD-L1/PD-1 blockade still shows slow response rates in controlling malignancies in many patients.Accumulating evidence has shown that an effective response to anti-PD-Ll/anti-PD-1 therapy requires establishing an integrated immune cycle.Damage in any step of the immune cycle is one of the most important causes of immunotherapy failure.Impairments in the immune cycle can be restored by epigenetic modification,including reprogramming the environment of tumor-associated immunity,eliciting an immune response by increasing the presentation of tumor antigens,and by regulating T cell trafficking and reactivation.Thus,a rational combination of PD-L1/PD-1 blockade and epigenetic agents may offer great potential to retrain the immune system and to improve clinical outcomes of checkpoint blockade therapy.

The regulatory and modulatory roles of TRP family channels in malignant tumors and relevant therapeutic strategies

Transient receptor potential(TRP)channels are one primary type of calcium(Ca^(2+))permeable channels,and those relevant transmembrane and intracellular TRP channels were previously thought to be mainly associated with the regulation of cardiovascular and neuronal systems.Nowadays,however,accumulating evidence shows that those TRP channels are also responsible for tumorigenesis and progression,inducing tumor invasion and metastasis.However,the overall underlying mechanisms and possible signaling transduction pathways that TRP channels in malignant tumors might still remain elusive.Therefore,in this review,we focus on the linkage between TRP channels and the significant characteristics of tumors such as multi-drug resistance(MDR),metastasis,apoptosis,proliferation,immune surveillance evasion,and the alterations of relevant tumor micro-environment.Moreover,we also have discussed the expression of relevant TRP channels in various forms of cancer and the relevant inhibitors’efficacy.The chemo-sensitivity of the anti-cancer drugs of various acting mechanisms and the potential clinical applications are also presented.Furthermore,it would be enlightening to provide possible novel therapeutic approaches to counteract malignant tumors regarding the intervention of calcium channels of this type.

cDNA cloning of four Hsp genes from Agarophyton vermiculophyllum and transcription analysis in different phases

Agarophyton vermiculophyllum is an agarophytic red alga originating from Northeast Asia that successfully spread to Europe and North America in the last 20 years.In this study,three hsp70 genes(hsp70-l,hsp70-2,and hsp70-3)and one hsp90 gene were cloned,and real-time reverse transcription polymerase chain reaction(RT-qPCR)was used to measure their transcriptional levels in three kinds of thalli(tetrasporophytes,and male/female gametophytes)belonging to diploid or haploid phases in the life cycle of A.vermiculophyllum.The results show that the three Hsp70s in A.vermiculophyllum clustered into three different groups,and the locations of the putative Hsp70-l,Hsp70-2,and Hsp70-3 were in the cytoplasm,endoplasmic reticulum and chloroplast,respectively,according to their sequences.Beside on the same research,the putative Hsp90 was supposed to have a cytoplasmic location.The RT-qPCR results show that the three hsp70 genes were highly upregulated in gametophytes as compared to tetrasporophytes but the transcriptional level of hsp90 did not show such a significant increase.The chloroplast hsp70-3 exhibited the highest upregulation and the transcriptional level in creased more than 570 fold in female gametophytes,and 17 fold in male gametophytes,compared with tetrasporophytes.Therefore,cpHsp70-3 might act more like a chaperon molecule involved in haploid development under natural condition,while Hsp70-l and Hsp70-2 were more active in stress resistance.

Enhanced modulation of magnetic field on surface plasmon coupled emission(SPCE) by magnetic nanoparticles

The obvious enhancement effect of magnetic nanoparticles(MNPs) introduced in Cr/Co/Cr/Au substrate on the pulsed magnetic field-modulated surface plasmon coupled emission(SPCE) was investigated,and the observed enhancement factor was 4 comparing with the magnetic field modulated SPCE without MNPs.This is the new observation for the magnetic field modulated SPCE,and this method was designed as a biosensor,which to our knowledge,is the first application of magnetic field-modulated SPCE in biosensing and detection field.This strategy is a universal approach to increase the fluorescence signal and helps to build the new SPCE based stimulus-response system.

Clinical characteristics and pre-surgery diagnostic criteria of aldosterone-producing adrenocortical carcinoma

To the Editor:Primary aldosteronism,characterized by hypertension,with or without hypokalemia,is the most frequent form of endocrine hypertension,which is mostly caused by bilateral adrenal hyperplasia or aldosterone-producing adrenocortical adenomas(APAA).However,it can also be induced by aldosterone-producing adrenocortical carcinoma(APAC),which is rare.Because of the rarity and malignancy.