Rapid detection of high-risk HPV16 and HPV18 based on microchip electrophoresis

Researches on detection of human papillomavirus(HPV)high-risk samples were carried out by polymerase chain reaction(PCR)coupled with microchip electrophoresis(MCE).Herein,we introduced a simple,rapid,automated method for detecting high-risk samples HPV16 and HPV18.In this research,general primers were initially selected to obtain sufficient detectable yield by PCR to verify feasibility of MCM method for HPV detection,then type-specific primers were further used to evaluate the specificity of MCE method.The results indicated MCE method was capable of specifically detecting high-risk HPV16 and HPV18,and also enabled simultaneous detection of multiplex samples.This MCE method described here has been successfully applied to HPV detection and displayed excellent reliability demonstrating by sequencing results.The inherent capability of MCE facilitated HPV detection conducted in a small chip with automated,high throughput,massive parallelized analysis.We envision that MCE method will definitely pave a way for clinical diagnosis,and even on-site screening of cervical cancer.

Photo‐catalyzed sequential dearomatization/carboxylation of benzyl o‐halogenated aryl ether with CO_(2) leading to spirocyclic carboxylic acids

Photo‐catalyzed tandem dearomatization/carboxylation of benzyl o‐halogenated aryl ether with CO_(2) was achieved,which affords spirocyclic carboxylic acids under mild conditions.The reaction has good functional group tolerance with high yields.Mechanism studies indicate that the transformation was realized via intramolecular radical addition and nucleophilic addition.

Matrix-assisted laser desorption ionization mass spectrometry based quantitative analysis of cordycepin from Cordyceps militaris

Cordycepin,which has great immunomodulatory activities such as anticancer,antifungal,antivirus,antileukemia and lipid-lowering ones,is the secondary metabolite of Cordyceps militaris(C.militaris).Liquid submerged fermentation is the common cultivation process to produce cordycepin.To optimize the fermentation process and improve production,monitoring the cordycepin secretion in the fermentation is essential.The measurement based on chromatography-mass spectrometry methods is generally involved in the complex sample pretreatments and time-consuming separation,so more rapid and convenient methods are required.Matrix-assisted laser desorption ionization mass spectrometry(MALDI-MS)is more attractive for faster and direct detection.Therefore,MALDI-MS detection combined with isotope-labeled internal standard was applied to the measurement of cordycepin content in the fermentation broth and mycelium.This method made accurate quantification of cordycepin in the range of 5-400μg/mL with a relative standard deviation of 5.6%.The recovery rates of fermentation samples after the 1,13,and 25 days were 90.15%,94.27%,and 95.06%,respectively.The contents of cordycepin in the mycelium and fermentation broth were 136 mg/g and 148.39 mg/mL on the 20 th culture day,respectively.The cordycepin secretion curve of the liquid fermentation of C.militaris was real-time traced over 25 days.

Recent progress in MOFs-based nanozymes for biosensing

Nanozymes are nanomaterials with enzyme-mimicking catalytic activity.Compared to natural enzymes,nanozymes show various properties such as easy to manufacture,stable,adjustable,and inexpensive.Nanozymes play key roles in biosensing,biocatalysis,and disease treatment.As an important kind of nanozymes,metal-organic framework(MOF)-based nanozymes are receiving a lot of attention due to their structural properties and composition.Rationally developing MOF with enzymes-like catalytic properties has opened new perspectives in biosensing.This review summarizes the up-to-date developments in synthesizing two-dimensional and three-dimensional MOF-based nanozymes and their applications in biosensing.Firstly,classification of nanozymes obtained by MOFs is categorized,and different properties of MOF-based nanozymes are described.Then,the distinctive applications of MOF-based nanozymes in identifying various analytes are thoroughly summarized.Finally,the recent challenges and progressive directions in this area are highlighted.

Efficient synthesis of cyclic imides by the tandem N-arylation-acylation and rearrangement reaction of cyanoesters with diaryliodonium salts

Available online An efficient method for the synthesis of multi-substituted cyclic imides was developed with cyanoesters and diaryliodonium salts.This method proceeds through a cascade of N-arylationacylation and rearrangement to give target heterocycles in good yields(up to 99%).This method has the major advantages of a broad substrate scope,excellent functional group compatibility.The strategy was also extended to the fused cyclic imides,such as malonimides,succinimides and glutarimides.

Discovery of small molecule degraders for modulating cell cycle

The cell cycle is a complex process that involves DNA replication,protein expression,and cell division.Dysregulation of the cell cycle is associated with various diseases.Cyclin-dependent kinases(CDKs)and their corresponding cyclins are major proteins that regulate the cell cycle.In contrast to inhibition,a new approach called proteolysis-targeting chimeras(PROTACs)and molecular glues can eliminate both enzymatic and scaffold functions of CDKs and cyclins,achieving targeted degradation.The field of PROTACs and molecular glues has developed rapidly in recent years.In this article,we aim to summarize the latest developments of CDKs and cyclin protein degraders.The selectivity,application,validation and the current state of each CDK degrader will be overviewed.Additionally,possible methods are discussed for the development of degraders for CDK members that still lack them.Overall,this article provides a comprehensive summary of the latest advancements in CDK and cyclin protein degraders,which will be helpful for researchers working on this topic.

Determining the contribution of Mo single atoms components in MoO_(2)nanocatalyst in transfer hydrogenation

Nanocatalysts are likely to contain undetected single-atom components,which may have been ignored but have significant effect in catalytic reactions.Herein,we report a catalyst composed of Mo single atoms(SAs)and MoO_(2)nanoparticles(NPs)(MoSAs-MoO_(2)@NC),which is an exact model to understand how the SAs contribute to the nanocatalyst.Both experimental results and the density functional theory calculations reveal that Mo SAs on nitrogen-doped carbon provides the reaction zone for nitro reduction,while MoO_(2)is the active site for decomposing hydrazine hydrate to produce H*.Thanks to the synergy between Mo SAs and MoO_(2)NPs,this catalyst exhibits noble metal-like catalytic activity(100%conversion at 4 min)for the dechlorination-proof transfer hydrogenation.Additionally,the hydrogen migration on the catalyst is verified by the electrochemical tests in the absence of a hydrogen source.This work provides a model for further study on the coexistence of single atoms in nanoparticle catalysts.

靶向难成药蛋白——蛋白质降解剂的力量

Undruggable targets typically refer to a class of therapeutic targets that are difficult to target through conventional methods or have not yet been targeted,but are of great clinical significance.According to statistics,over 80%of disease-related pathogenic proteins cannot be targeted by current conventional treatment methods.In recent years,with the advancement of basic research and new technologies,the development of various new technologies and mechanisms has brought new perspectives to overcome challenging drug targets.Among them,targeted protein degradation technology is a breakthrough drug development strategy for challenging drug targets.This technology can specifically identify target proteins and directly degrade pathogenic target proteins by utilizing the inherent protein degradation pathways within cells.This new form of drug development includes various types such as proteolysis targeting chimera(PROTAC),molecular glue,lysosome-targeting Chimaera(LYTAC),autophagosometethering compound(ATTEC),autophagy-targeting chimera(AUTAC),autophagy-targeting chimera(AUTOTAC),degrader-antibody conjugate(DAC).This article systematically summarizes the application of targeted protein degradation technology in the development of degraders for challenging drug targets.Finally,the article looks forward to the future development direction and application prospects of targeted protein degradation technology.

Distance-basedα-amylase biosensor fabricated with amylopectin-coated mesoporous membrane

Paper-based biosensors are widely employed in point-of-care testing(POCT)due to their convenience,portability,low cost,and ease of use.This study reports an integrated distance-based paper biosensor fabricated with a mesoporous membrane coated with stimuli-responsive polymer.The detection ofα-amylase(AMY)using amylopectin-coated mesoporous membrane is demonstrated as an example.After introducing the AMY solution,it is observed that the aqueous solution flows along the paper strip due to AMY-catalyzed hydrolysis of amylopectin.The flow distance is proportional to the concentration of AMY with a detection limit as low as 4 mU/mL.In addition,the detection of AMY is demonstrated in human serum.Furthermore,the inhibitory effect of acarbose on AMY is evaluated.This reagent-free and disposable biosensor allows single-step rapid detection of the analyte.This approach is very promising for the development of user-friendly,equipment-free,and cost-effective biosensors with remarkable sensitivity and excellent selectivity for disease diagnosis and hypoglycemic drug screening.

Microfluidic adhesion analysis of single glioma cells for evaluating the effect of drugs

Cancer metastasis is one of the most serious problems for tumor therapy,which is closely related to cell adhesion and deadhesion process.Better comprehension of cell adhesion ability will benefit drug research.Here,a biomimetic microfluidic enzyme digestion method was proposed to gently measure the influence of drugs on cell-matrix adhesion ability at the single cell level.The method can selectively digest the extracellular matrix(ECM)that linked to a single cell,and the trypsin concentration around the cell is relatively uniform and constant,thus the measured cell adhesion strength should be precise.Commercially available anti-cancer agents including 5-fluorouracil(5-FU),actinomycin D(Act D),temozolomide(TMZ)and allicin were evaluated,and the data showed only TMZ and allicin can inhibit cell adhesion significantly under our experiment conditions.The influence of TMZ became more and more obvious as the increase of duration and the effect became prominent only after 6 h adhesion process,which could provide a quick evaluation of whether the drugs are effective to cancer cell(compared with Calcein-AM/PI cell viability test).The adhesion strength of U87 cells decreased when the concentration of TMZ increased,and the effect of TMZ can be effectively inhibited by adding lactic acid to culture medium,which indicated acidic tumor microenvironment could promote drug resistance of tumor cells.Different from conventional evaluation methods which focus on the drugs’influence on cellular viability or metabolism,this work provides a new perspective to study the effect of drugs,which is helpful to enrich the drug evaluation system.

Correction to: Developing potent PROTACs tools for selective degradation of HDAC6 protein

In the original publication the title of X axis in Fig.1G is in correctly published as"Compound(pmol/L)".The correct title of X axis in Fig.1G should be read as"Compound(nmol/L)M.Figure 1.Development of selective HDAC6-degrading PROTACs.(A)The principle of PROTAC.(B)The structure of PROTAC,as shown in the upper portion.A binding mode of PROTAC(ball stick),HDAC6(PDB 5G0J,purple)and CRL4-CRBN(PDB 2HYE and 4CI3,colored cyan and gray)was simulated by Pymol.

Genome-wide evolution analysis reveals low CpG contents of fast-evolving genes and identifies antiviral microRNAs

Noncoding RNAs(ncRNAs) play important roles in many biological processes and provide materials for evolutionary adaptations beyond protein-coding genes, such as in the arms race between the host and pathogen. However, currently, a comprehensive high-resolution analysis of primate genomes that includes the latest annotated ncRNAs is not available. Here, we developed a computational pipeline to estimate the selections that act on noncoding regions based on comparisons with a large number of reference sequences in introns adjacent to the interested regions. Our method yields result comparable with those of the established codon-based method and phyloP method for coding genes;thus, it provides a holistic framework for estimating the selection on the entire genome. We further showed that fastevolving protein-coding genes and their corresponding 50 UTRs have a significantly lower frequency of the CpG dinucleotides than those evolving at an average pace, and these fast-evolving genes are enriched in the process of immunity and host defense. We also identified fast-evolving miRNAs with antiviral functions in cells. Our results provide a resource for high-resolution evolution analysis of the primate genomes.

Design,synthesis,and biological evaluation of multiple targeting antimalarials

Malaria still threatens global health seriously today.While the current discoveries of antimalarials are almost totally focused on single mode-of-action inhibitors,multi-targeting inhibitors are highly desired to overcome the increasingly serious drug resistance.Here,we performed a structure-based drug design on mitochondrial respiratory chain of Plasmodium falciparum and identified an extremely potent molecule,RYL-581,which binds to multiple protein binding sites of P.falciparum simultaneously(allosteric site of type Ⅱ NADH dehydrogenase,Q_(o) and Q_(i) sites of cytochrome bc_(1)).Antimalarials with such multiple targeting mechanism of action have never been reported before.RYL-581 kills various drug-resistant strains in vitro and shows good solubility as well as in vivo activity.This structurebased strategy for designing RYL-581 from starting compound may be helpful for other medicinal chemistry projects in the future,especially for drug discovery on membrane-associated targets.

Shell microparticles of morphology controlled and inner-modified hole from sequential inkjet-printed double emulsions

This article presents sequential inkjet-based method to produce double emulsions as templates to fabricate morphology-controlled and inner-modified hole-shell microparticles. This sequential printing strategy for producing double emulsions circumvents complex wettability modification of the microchannels in lithography-based microfluidic device and largely saves the reagent in comparison to the coaxial two-phase jet in glass capillary. The formation of hole-shell structures is attributed to the diffusion of solvent out of droplets into butanol at the interface between oil and extract phase. The change of hole size is controlled by different diffusion rate, which is determined by changing the volume ratio of butanol and alcohol in extract phase.This presented flexible method can fabricate some functionalized microparticles in our future work.

MeOTf-catalyzed formal[4+2]annulations of styrene oxides with alkynes leading to polysubstituted naphthalenes through sequential electrophilic cyclization/ring expansion

Me OTf-catalyzed formal[4+2]annulation of styrene oxides with alkynes to afford polysubstituted naphthalenes has been realized,which undergoes sequential electrophilic cyclization/ring expansion.A range of substrates were tolerated in the formation of naphthalene derivatives with high regioselectivity in satisfactory yields.The reaction could also be carried out on gram scale.

Di-4-ANEPPDHQ probes the response of lipid packing to the membrane tension change in living cells

Membrane tension plays a significant role in many cellular processes including cell adhesion, migration and spreading. Despite the importance of membrane tension, it remains difficult to measure in vivo. Recently, the development of non-invasive fluorescent probes have made great progress, especially excitedstate deplanarization in molecular rotors has been applied to image membrane tension in living cells.Nevertheless, an intrinsic limitation of such kind of probe is that they depend on the lipid packing, and how the lipid packing responds to the membrane tension change remains unclear. Therefore, in this work,we used a polarity-sensitive membrane probe to investigate the possible response mechanism of lipid packing to the change of membrane tension that was regulated by osmotic shocks. The results showed that an increase in membrane tension could stretch the lipids apart with large displacements, and this change was not homogeneous on the whole membrane, instead, increase of membrane tension induced phase separation.

Elucidating the selectivity of dyotropic rearrangements of β-lactones:a computational survey

The dyotropic rearrangement of β-lactones is a neglected treasure in the family of multi-bond reactions and pericyclic reactions.Despite its appealing synthetic potential,the complicated migration selectivity greatly limits its widespread application.In this work,we report the first systematic and comprehensive computational study on the dyotropic rearrangements of β-lactones.The use of the double-hybrid functional ensures the accuracy of results.On the basis of the present study and our previous work,five methods to control the reaction selectivity of dyotropic rearrangements of β-lactones have been summarized,providing valuable references for synthetic chemists to design and develop brand-new type dyotropic reactions.

Synthesis of Quaternary Carbon-Centered Benzoindolizidinones via Novel Photoredox-Catalyzed Alkene Aminoarylation: Facile Access to Tylophorine and Analogues

Photoredox-catalyzed aminoarylation and thioami-nation of unactivated alkenes have been developed,providing novel synthetic routes to access synthe-tically challenging quaternary carbon-centered benzoindolizidinones and trifluoromethylthiolated piperidines using readily available starting materials.Notably,these transformations were enabled by merging amidyl radical generation from N-alkyl benzamides with oxidant incorporation.Density functional theory calculations were performed to understand the reaction mechanism and to rationa-lize the regioselectivities.Moreover,the newly deve-loped catalytic aminoarylation provided a convenient synthetic route for natural product tylophorine and its gem-dimethyl analogues with greatly improved drug-like properties such as enhanced solubility and stability.

ASFV transcription reporter screening system identifies ailanthone as a broad antiviral compound

African swine fever(ASF)is an acute,highly contagious and deadly viral disease in swine that jeopardizes the worldwide pig industry.Unfortunately,there are no authoritative vaccine and antiviral drug available for ASF control.African swine fever virus(ASFV)is the etiological agent of ASF.Among the ASFV proteins,p72 is the most abundant component in the virions and thus a potential target for anti-ASFV drug design.Here,we con-structed a luciferase reporter system driven by the promoter of p72,which is transcribed by the co-transfected ASFV RNA polymerase complex.Using this system,we screened over 3200 natural product compounds and obtained three potent candidates against ASFV.We further evaluated the anti-ASFV effects and proved that among the three candidates,ailanthone(AIL)inhibits the replication of ASFV at the nanomolar concentration(IC_(50)=15 nmol/L).Our in vitro experiments indicated that the antiviral effect of AIL is associated with its inhibition of the HSP90-p23 cochaperone.Finally,we showed the antiviral activity of AIL on Zika virus and hepatitis B virus(HBV),which supports that AIL is a potential broad-spectrum antiviral agent.

Developing potent BTK^(C481S)PROTACs for ibrutinib-resistant malignant lymphoma

Ibrutinib is a first-line treatment drug for B-cell malignancies.However,resistance to ibrutinib has been reported due to BTKC481Smutation.Although PROTAC strategy is expected to overcome this clinical resistance,it has limitations such as large molecular weight and moderate bioactivity,which restrict its potential clinical application.Herein,we report a new type of potent BTKC481S-targeting PROTAC degrader.Through design,computer-assisted optimization and SAR studies,we have developed a representative BTKC481Sdegrader L6 with a much smaller molecular weight and improved solubility.Notably,L6 demonstrates better BTK degrading activity and lower IC50value in ibrutinib-resistant cell line than the first-generation BTK degrader P13I.Optimization strategy of L6 provides a general approach in the development of PROTACs targeting BTK and other proteins for future study.