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-st...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.展开更多
Agrobacterium tumefaciens-mediated DNA transiormation method was applied to transform Noclulisporium sylviforme fusant HDF-68, a taxol-produeing fungus. We constructed a binary vector pBI121-43 canting a hygromycin-re...Agrobacterium tumefaciens-mediated DNA transiormation method was applied to transform Noclulisporium sylviforme fusant HDF-68, a taxol-produeing fungus. We constructed a binary vector pBI121-43 canting a hygromycin-resistant gene cassette between the right and left borders of T-DNA, Optimal co-cultivation of N.sylviforrne with A. tumefaciens containing pBI121-43 led to 110- 130 hygromycin-resistant transformants per＂ million eonidia. Putative transformants were found to be mitotically stable. The molecular analysis of transformants demonstrated the random integration of single copy of the T-DNA into the host genome. This transformation system serves as a basic tool for insertional mutagenesis in N. sylviforme fusant HDF-68, and the development of such svstem lays a solid foundation for constructing high-yied gene engineering strain and clarifying taxol biosynthesis pathway in this fungus.展开更多
Comprehensive understanding of mechanisms of epigenetic regulation requires identification of molecules bound to genomic regions of interest in vivo. We have developed a novel method, insertional chromatin immunopreci...Comprehensive understanding of mechanisms of epigenetic regulation requires identification of molecules bound to genomic regions of interest in vivo. We have developed a novel method, insertional chromatin immunoprecipitatin (iChIP), to isolate specific genomic regions retaining molecular interaction in order to perform non-biased identification of interacting molecules in vivo. Here, we developed a second-generation tagged LexA DNA-binding domain, 3xFNLDD, for the iChIP analysis. 3xFNLDD consists of 3 x FLAG tags, a nuclear localization signal (NLS), the DNA-binding domain (DB) and the dimerization domain of the LexA protein. Expression of 3xFNLDD can be detected by immunoblot analysis as well as flowcytometry. We showed that iChIP using 3xFNLDD is able to consistently isolate more than 10% of input genomic DNA, several-fold more efficient compared to the first-generation tagged LexA DB. 3xFNLDD would be a useful tool to perform the iChIP analysis for locus-specific biochemical epigenetics.展开更多
Background: Treatment of insertional achilles tendinopathy is known to be difficult. Eccentric calf muscle exercises and extracorporeal shockwave therapy have been proposed as the primary treatments for this condition...Background: Treatment of insertional achilles tendinopathy is known to be difficult. Eccentric calf muscle exercises and extracorporeal shockwave therapy have been proposed as the primary treatments for this condition, but surgery is indicated after failed conservative management. There is no consensus about the most efficient surgical treatment. Objectives: To evaluate a new ultrasound and color Doppler-guided surgical treatment for insertional achilles tendinopathy. Patients and methods: 24 consecutive patients (13 men and 11 women, mean age 47 years) with a long duration of insertional achilles tendon pain (median 18 months) were included in the study. The surgical procedure consisted of extirpation of the subcutaneous and retrocalcaneal bursa, scraping of the ventral distal achilles tendon and removal of prominent bone at the upper calcaneal tuberosity (Haglund’s deformity). In 13 patients, the plantaris tendon was also cut and excised. VAS for pain during activity, satisfaction with the treatment result and the SF-36 score to measure quality of life were used for evaluation. Results: The mean VAS had decreased from 72 before surgery to 19 after surgery (p 0.001), and 20/24 (83%) patients were satisfied with the result and had returned to full tendon loading activity at the one-year follow-up. The SF-36 score had improved significantly. Conclusion: Ultrasound and color Doppler-guided surgical treatment seems to be a good method for treatment of chronic painful insertional achilles tendinopathy. Longer follow-up studies on larger materials are needed.展开更多
In plants,transposable element(TE)-triggered mutants are important resources for functional genomic studies.However,conventional approaches for genome-wide identification of TE insertion sites are costly and laborious...In plants,transposable element(TE)-triggered mutants are important resources for functional genomic studies.However,conventional approaches for genome-wide identification of TE insertion sites are costly and laborious.This study developed a novel,rapid,and high-throughput TE insertion site identification workflow based on next-generation sequencing and named it Transposable Element Amplicon Sequencing(TEAseq).Using TEAseq,we systemically profiled the Dissociation(Ds)insertion sites in 1606 independent Ds insertional mutants in advanced backcross generation using K17 as background.The Ac-containing individuals were excluded for getting rid of the potential somatic insertions.We characterized 35,696 germinal Ds insertions tagging 10,323 genes,representing approximately 23.3%of the total genes in the maize genome.The insertion sites were presented in chromosomal hotspots around the ancestral Ds loci,and insertions occurred preferentially in gene body regions.Furthermore,we mapped a loss-of-function AGL2 gene using bulked segregant RNA-sequencing assay and proved that AGL2 is essential for seed development.We additionally established an open-access database named MEILAM for easy access to Ds insertional mutations.Overall,our results have provided an efficient workflow for TE insertion identification and rich sequence-indexed mutant resources for maize functional genomic studies.展开更多
Three T-DNA insertional embryonic lethal mutants from NASC (The Nottingham Arabidopsis Stock Center)were first checked with their segregation ratio of abortive and normal seeds and the copy number of T-DNA insertion. ...Three T-DNA insertional embryonic lethal mutants from NASC (The Nottingham Arabidopsis Stock Center)were first checked with their segregation ratio of abortive and normal seeds and the copy number of T-DNA insertion. The N4081 mutant has a segregation ratio of 1:3.04in average and one T-DNA insertion site according to our assay It was therefore chosen for further analysis. To isolate the joint fragment of T-DNA and plan DNA, the plasmid rescue technique waJs used. pEL-7, one of plasmids from left border of T-DNA, which contained pBR322 was selected from ampicillin plate. The T-DNA fragment of pEL-7 was checked by restriction enzyme analysis and Southern Blot. Restriction analysis confirmed the presence of known sites of EcoRI, PstI and PvuII on it.For confirming the presence of flanking plant DNA in this plasmid, pEL-7 DNA was labeled and hybridized with wild type and mutant plant DNA. The Southern Blot indicated the hybridization band in both of them. Furthermore, the junction of T-DNA/plant DNA was subcloned into bluescript SK+ and sequenced by Applied Biosystem 373A Sequencer. The results showed the 822 bp fragment contained a 274 bp sequence, which is 99.6%homolog (273bp/274 bp) to Ti plasmid pTi 15955 DNA.Ten bp of left 25 bp border repeat were also found in the juction of T-DNA and Plant DNA.Taken together, pEL-7 should contain a joint fragment of T-DNA and flanking plant DNA. This plasmid DNA could be used for the isolation of plant gene, which will be helpful to elucidate the relationship between gene function and plant embryo development.展开更多
Mutator transposable element （Mu） has been used as an effective tool to clone maize （Zea mays L.） genes. One opaque endosperm mutant （miol6） was identified in a pool of Mu inserted mutants. A modified method, te...Mutator transposable element （Mu） has been used as an effective tool to clone maize （Zea mays L.） genes. One opaque endosperm mutant （miol6） was identified in a pool of Mu inserted mutants. A modified method, termed the double selected amplification of insertion flanking fragments （DSAIFF）, was employed to isolate the Mu flanking fragments （MFFs） of miol6. The target site duplications （TSDs） isolated from the Msp I and Mse I digested MFFs had a same 9-bp sequence and were confirmed to be the flanking sequence of one identically inserted gene. Co-segregation analysis suggested that the MFFs were associated with the mutant opaque endosperm, and miol6 was mapped in silico onto the physical position ranged from 229 965 021 to 229 965 409 bp of the maize chromosome 4.09 bin. The full-length cDNA of the wild-type gene was obtained by an RT-PCR primer-scanning technique, and Mio16 was found to putatively encode a homolog of the Arabidopsis MAP3K delta-1 protein kinase. RT-PCR result the mRNA expression of miol6 region anchored by primers Mu20 and af276 was not interrupted by Mu insertion. Further researches will be done to elucidate how the expression of miol6 is alternated by Mu insertion.展开更多
BACKGROUND Long-term chemotherapy for patients with gastric cancer(GC),facilitated by peripherally inserted central catheter(PICC)catheterization,reduces vascular damage and enhances drug delivery efficiency but carri...BACKGROUND Long-term chemotherapy for patients with gastric cancer(GC),facilitated by peripherally inserted central catheter(PICC)catheterization,reduces vascular damage and enhances drug delivery efficiency but carries risks of catheter-related complications.A combination of group psychological nursing and physical mo-vement care significantly mitigates the risk of venous thrombosis and improves psychological well-being,and enhances motor function,underscoring its clinical importance.AIM To assess group psychological and physical movement nursing in preventing venous thrombosis in patients with PICC GC.METHODS Sixty-five GC patients with PICC,admitted from January 2022 to January 2023,were randomly divided into two groups using the lottery method:A control group(n=35,routine nursing)and an observation group(n=30,routine nursing plus psychological nursing and physical movement nursing).Both groups re-ceived continuous care for 2 weeks.Pre-nursing and post-nursing data on psycho-logical state,physical function,chemotherapy-related thrombosis incidence,and cancer-related fatigue were analyzed using SPSS 26.0 and GraphPad Prism 8.0.RESULTS After nursing,both groups showed reduced Hamilton Anxiety Scale scores and increased General Perceived Self-Efficacy Scale scores,with the observation group performing better(P<0.05).The Functional Comprehensive Assessment score for the observation group after nursing was(65.42±2.35)points,lower than the control group’s(62.19±4.33)points(P<0.05).Although no significant difference was observed in the incidence of venous thrombosis between the two groups(χ2=0.815,P=0.367),the observation group had lower incidence.Both groups showed decreased Revised Piper Fatigue Scale scores,with the observation group scoring lower(P<0.05).CONCLUSION Group psychological and physical movement nursing for patients with PICC reduces venous thrombosis risk,improves psychological well-being,cancer-related fatigue,and physical function,making it highly promotable.展开更多
An efficient TfOH-catalyzed O—H insertion reaction of α-aryl diazoesters with carboxylic acids is reported.This metal-free protocol provides an operationally simple method for a one-pot assembly of diverse α-acylox...An efficient TfOH-catalyzed O—H insertion reaction of α-aryl diazoesters with carboxylic acids is reported.This metal-free protocol provides an operationally simple method for a one-pot assembly of diverse α-acyloxy esters in moderate to high yields with a broad substrate scope.All starting materials are readily available,and the reactions can be conducted in the open air at room temperature.展开更多
A blue light-induced formal insertion reaction ofα-siloxy carbene into the C—H bond of 1,3-diketones has been reported.Under the irradiation of blue light,acylsilane converts toα-siloxy carbene,which then undergoes...A blue light-induced formal insertion reaction ofα-siloxy carbene into the C—H bond of 1,3-diketones has been reported.Under the irradiation of blue light,acylsilane converts toα-siloxy carbene,which then undergoes formal C—H bond insertion reaction with the enol form of 1,3-diketone.This method uses readily available and relative stable acylsilane as car-bene precursor,which features a simple and metal-free approach under mild conditions.Moreover,the synthetic potential of this protocol has been demonstrated by performing the reaction on a gram scale with comparable high yield.展开更多
There is nonradiative recombination in waveguide region owing to severe carrier leakage,which in turn reduces output power and wall-plug efficiency.In this paper,we designed a novel epitaxial structure,which suppresse...There is nonradiative recombination in waveguide region owing to severe carrier leakage,which in turn reduces output power and wall-plug efficiency.In this paper,we designed a novel epitaxial structure,which suppresses carrier leakage by inserting n-Ga_(0.55)In_(0.45)P and p-GaAs_(0.6)P_(0.4) between barriers and waveguide layers,respectively,to modulate the energy band structure and to increase the height of barrier.The results show that the leakage current density reduces by 87.71%,compared to traditional structure.The nonradiative recombination current density of novel structure reduces to 37.411 A/cm^(2),and the output power reaches 12.80 W with wall-plug efficiency of 78.24%at an injection current density 5 A/cm^(2) at room temperature.In addition,the temperature drift coefficient of center wavelength is 0.206 nm/℃at the temperature range from 5℃to 65℃,and the slope of fitted straight line of threshold current with temperature variation is 0.00113.The novel epitaxial structure provides a theoretical basis for achieving high-power laser diode.展开更多
Virus-based vectors are widely used in hematopoietic stem cell(HSC)gene therapy,and have the ability to integrate permanently into genomic DNA,thus driving long-term expression of corrective genes in all hematopoietic...Virus-based vectors are widely used in hematopoietic stem cell(HSC)gene therapy,and have the ability to integrate permanently into genomic DNA,thus driving long-term expression of corrective genes in all hematopoietic lineages.To date,HSC gene therapy has been successfully employed in the clinic for improving clinical outcomes in small numbers of patients with X-linked severe combined immunodeficiency(SCID-X1),adenosine deaminase deficiency(ADA-SCID),adrenoleukodystrophy(ALD),thalassemia,chronic granulomatous disease(CGD),and Wiskott-Aldrich syndrome(WAS).However,adverse events were observed during some of these HSC gene therapy clinical trials,linked to insertional activation of proto-oncogenes by integrated proviral vectors leading to clonal expansion and eventual development of leukemia.Numerous studies have been performed to understand the molecular basis of vector-mediated genotoxicity,with the aim of developing safer vectors and lower-risk gene therapy protocols.This review will summarize current information on the mechanisms of insertional mutagenesis in hematopoietic stem and progenitor cells due to integrating gene transfer vectors,discuss the available assays for predicting genotoxicity and mapping vector integration sites,and introduce newlydeveloped approaches for minimizing genotoxicity as a way to further move HSC gene therapy forward into broader clinical application.展开更多
The switchable cross-coupling of indoles and pyridotriazoles through carbene insertion at C_(2)-or C_(3)-positon has been developed in this paper.This highly site-selective C-H carbenoid functionalization is determine...The switchable cross-coupling of indoles and pyridotriazoles through carbene insertion at C_(2)-or C_(3)-positon has been developed in this paper.This highly site-selective C-H carbenoid functionalization is determined by both the Rh-catalyst species and auxiliary groups.[Cp∗RhCl_(2)]_(2) and coordinating pyrimidyl group direct the C-H carbenoid functionalization to occur at the C_(2)-position,while Rh2OAc4 and noncoordinating benzyl group lead the reaction to occur at the C_(3)-position of the indoles.This regioselective C−H functionalization strategy is of significant importance for the discovery of indole drugs.展开更多
Achieving high-performance aqueous zinc-ion batteries requires addressing the challenges associated with the stability of zinc metal anodes,particularly the formation of inhomogeneous zinc dendrites during cycling and...Achieving high-performance aqueous zinc-ion batteries requires addressing the challenges associated with the stability of zinc metal anodes,particularly the formation of inhomogeneous zinc dendrites during cycling and unstable surface electrochemistry.This study introduces a practical method for scattering untreated bulk hexagonal boron nitride(h-BN)particles onto the zinc anode surface.During cycling,stabilized zinc fills the interstices of scattered h-BN,resulting in a more favorable(002)orientation.Consequently,zinc dendrite formation is effectively suppressed,leading to improved electrochemical stability.The zinc with scattered h-BN in a symmetric cell configuration maintains stability 10 times longer than the bare zinc symmetric cell,lasting 500 hours.Furthermore,in a full cell configuration withα-MnO_(2) cathode,increased H^(+)ion activity can effectively alter the major redox kinetics of cycling due to the presence of scattered h-BN on the zinc anode.This shift in H^(+)ion activity lowers the overall redox potential,resulting in a discharge capacity retention of 96.1%for 300 cycles at a charge/discharge rate of 0.5 Ag^(-1).This study highlights the crucial role of surface modification,and the innovative use of bulk h-BN provides a practical and effective solution for improving the performance and stability.展开更多
Avian metapneumovirus(aMPV),a paramyxovirus,causes acute respiratory diseases in turkeys and swollen head syndrome in chickens.This study established a reverse genetics system for aMPV subtype B LN16-A strain based on...Avian metapneumovirus(aMPV),a paramyxovirus,causes acute respiratory diseases in turkeys and swollen head syndrome in chickens.This study established a reverse genetics system for aMPV subtype B LN16-A strain based on T7 RNA polymerase.Full-length cDNA of the LN16-A strain was constructed by assembling 5 cDNA fragments between the T7 promoter and hepatitis delta virus ribozyme.Transfection of this plasmid,along with the supporting plasmids encoding the N,P,M2-1,and L proteins of LN16-A into BSR-T7/5 cells,resulted in the recovery of aMPV subtype B.To identify an effective insertion site,the enhanced green fluorescent protein(EGFP)gene was inserted into different sites of the LN16-A genome to generate recombinant LN16-As.The results showed that the expression levels of EGFP at the site between the G and L genes of LN16-A were significantly higher than those at the other two sites(between the leader and N genes or replacing the SH gene).To verify the availability of the site between G and L for foreign gene expression,the VP2 gene of very virulent infectious bursal disease virus(vvIBDV)was inserted into this site,and recombinant LN16-A(rLN16A-vvVP2)was successfully rescued.Single immunization of specificpathogen-free chickens with rLN16A-vvVP2 induced high levels of neutralizing antibodies and provided 100%protection against the virulent aMPV subtype B and vvIBDV.Establishing a reverse genetics system here provides an important foundation for understanding aMPV pathogenesis and developing novel vector vaccines.展开更多
The ThinKing by Russian watchmaker Konstantin Chaykin is only 1.65 millimeters thick,making it the world's thinnest mechanical wristwatch.The“arms race”for the world's thinnest wristwatch has been in full sw...The ThinKing by Russian watchmaker Konstantin Chaykin is only 1.65 millimeters thick,making it the world's thinnest mechanical wristwatch.The“arms race”for the world's thinnest wristwatch has been in full swing for a few years now,with some of the most prestigious(有声望的)watch brands on Earth trying to one‑up each other with ingenious designs.The latest to claim the coveted title is independent watchmaker Konstantin Chaykin with his ThinKing prototype,a 1.65‑millimeter‑thick masterpiece that is so incredibly thin that it needs a special wristband.The ThinKing's case is so delicate that the simple pressure from being worn is enough to damage it,so the special strap made of alligator leather and elastic(有弹性的)inserts with titanium(钛)has to absorb some of the shocks,twisting and bending.展开更多
Intelligent production is an important development direction in intelligent manufacturing,with intelligent factories playing a crucial role in promoting intelligent production.Flexible job shops,as the main form of in...Intelligent production is an important development direction in intelligent manufacturing,with intelligent factories playing a crucial role in promoting intelligent production.Flexible job shops,as the main form of intelligent factories,constantly face dynamic disturbances during the production process,including machine failures and urgent orders.This paper discusses the basic models and research methods of job shop scheduling,emphasizing the important role of dynamic job shop scheduling and its response schemes in future research.A multi-objective flexible job shop dynamic scheduling mathematical model is established,highlighting its complex and multi-constraint characteristics under different interferences.A classification discussion is conducted on the dynamic response methods and optimization objectives under machine failures,emergency orders,fuzzy completion times,and mixed dynamic events.The development process of traditional scheduling rules and intelligent methods in dynamic scheduling are also analyzed.Finally,based on the current development status of job shop scheduling and the requirements of intelligent manufacturing,the future development trends of dynamic scheduling in flexible job shops are proposed.展开更多
Uronic acids are prevalent components of crucial glycoconjugates,pivotal in various biological processes.In nature,NDP-uronic acids,the nucleosides-activated uronic acids,serve as glycosylation donors catalyzed by uro...Uronic acids are prevalent components of crucial glycoconjugates,pivotal in various biological processes.In nature,NDP-uronic acids,the nucleosides-activated uronic acids,serve as glycosylation donors catalyzed by uronosyltransferases(UATs)to construct glycans containing uronic acids.Despite their biological importance,the synthesis of naturally occurring NDP-uronic acids on a large scale remains challenging.Here,we developed an oxidation reaction insertion strategy for the efficient synthesis of NDP-uronic acids,and 11 NDP-uronic acids were successfully prepared in good yield and on a large scale.The prepared NDP-uronic acids can be used to explore new uronosyltransferases and synthesize uronic acids containing carbohydrates for fundamental research.展开更多
Genome rearrangement is an important process that leads to genetic diversity,including mutation-related insertions,deletions,or inversions in the genome[1,2].
BACKGROUND Peripherally inserted central catheters(PICCs)are widely used for administering chemotherapy to breast cancer patients due to their long-term indwelling capability,versatility in drug administration,and fle...BACKGROUND Peripherally inserted central catheters(PICCs)are widely used for administering chemotherapy to breast cancer patients due to their long-term indwelling capability,versatility in drug administration,and flexibility.PICCs infection are a relatively common occurrence,yet there were no reported instances that it can metastasise to the lumbar spine.CASE SUMMARY This case report describes a breast cancer patient who developed a methicillinresistant Staphylococcus aureus lumbar vertebral infection secondary to a PICCrelated infection during chemotherapy.Following PICC removal,bacterial culture confirmed the presence of highly virulent methicillin-resistant Staphylococcus aureus.The patient presented with fever and severe lumbar pain.Lumbar magnetic resonance imaging revealed paraspinal muscle edema from L1 to L3 with abnormal signal intensity in the affected regions,suggestive of vertebral osteomyelitis.Prompt initiation of appropriate antibiotic therapy based on the culture results led to significant improvement in the patient’s lumbar pain.CONCLUSION This case highlights the importance of vigilant infection prevention and control measures to minimize the risk of PICC-related complications,such as bloodstream infections and subsequent metastatic infections.展开更多
基金the National Key R&D Program of China(Nos.2018YFD0901506,2018YFD0900305)the Marine S&T Fund of Shandong Province for Pilot National Laboratory for Marine Science and Technology(Qingdao)(No.2018 SDKJ0406-3)。
文摘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.
基金the National Natural Science Foundation of China(No.30570025)the Education Department of Heilongjiang Province(No.10551238)
文摘Agrobacterium tumefaciens-mediated DNA transiormation method was applied to transform Noclulisporium sylviforme fusant HDF-68, a taxol-produeing fungus. We constructed a binary vector pBI121-43 canting a hygromycin-resistant gene cassette between the right and left borders of T-DNA, Optimal co-cultivation of N.sylviforrne with A. tumefaciens containing pBI121-43 led to 110- 130 hygromycin-resistant transformants per＂ million eonidia. Putative transformants were found to be mitotically stable. The molecular analysis of transformants demonstrated the random integration of single copy of the T-DNA into the host genome. This transformation system serves as a basic tool for insertional mutagenesis in N. sylviforme fusant HDF-68, and the development of such svstem lays a solid foundation for constructing high-yied gene engineering strain and clarifying taxol biosynthesis pathway in this fungus.
文摘Comprehensive understanding of mechanisms of epigenetic regulation requires identification of molecules bound to genomic regions of interest in vivo. We have developed a novel method, insertional chromatin immunoprecipitatin (iChIP), to isolate specific genomic regions retaining molecular interaction in order to perform non-biased identification of interacting molecules in vivo. Here, we developed a second-generation tagged LexA DNA-binding domain, 3xFNLDD, for the iChIP analysis. 3xFNLDD consists of 3 x FLAG tags, a nuclear localization signal (NLS), the DNA-binding domain (DB) and the dimerization domain of the LexA protein. Expression of 3xFNLDD can be detected by immunoblot analysis as well as flowcytometry. We showed that iChIP using 3xFNLDD is able to consistently isolate more than 10% of input genomic DNA, several-fold more efficient compared to the first-generation tagged LexA DB. 3xFNLDD would be a useful tool to perform the iChIP analysis for locus-specific biochemical epigenetics.
文摘Background: Treatment of insertional achilles tendinopathy is known to be difficult. Eccentric calf muscle exercises and extracorporeal shockwave therapy have been proposed as the primary treatments for this condition, but surgery is indicated after failed conservative management. There is no consensus about the most efficient surgical treatment. Objectives: To evaluate a new ultrasound and color Doppler-guided surgical treatment for insertional achilles tendinopathy. Patients and methods: 24 consecutive patients (13 men and 11 women, mean age 47 years) with a long duration of insertional achilles tendon pain (median 18 months) were included in the study. The surgical procedure consisted of extirpation of the subcutaneous and retrocalcaneal bursa, scraping of the ventral distal achilles tendon and removal of prominent bone at the upper calcaneal tuberosity (Haglund’s deformity). In 13 patients, the plantaris tendon was also cut and excised. VAS for pain during activity, satisfaction with the treatment result and the SF-36 score to measure quality of life were used for evaluation. Results: The mean VAS had decreased from 72 before surgery to 19 after surgery (p 0.001), and 20/24 (83%) patients were satisfied with the result and had returned to full tendon loading activity at the one-year follow-up. The SF-36 score had improved significantly. Conclusion: Ultrasound and color Doppler-guided surgical treatment seems to be a good method for treatment of chronic painful insertional achilles tendinopathy. Longer follow-up studies on larger materials are needed.
基金the Ministry of Science and Technology of China(2016YFD0101000 and 2016YFD0101001)the Natural Science Foundation of China(31901595).
文摘In plants,transposable element(TE)-triggered mutants are important resources for functional genomic studies.However,conventional approaches for genome-wide identification of TE insertion sites are costly and laborious.This study developed a novel,rapid,and high-throughput TE insertion site identification workflow based on next-generation sequencing and named it Transposable Element Amplicon Sequencing(TEAseq).Using TEAseq,we systemically profiled the Dissociation(Ds)insertion sites in 1606 independent Ds insertional mutants in advanced backcross generation using K17 as background.The Ac-containing individuals were excluded for getting rid of the potential somatic insertions.We characterized 35,696 germinal Ds insertions tagging 10,323 genes,representing approximately 23.3%of the total genes in the maize genome.The insertion sites were presented in chromosomal hotspots around the ancestral Ds loci,and insertions occurred preferentially in gene body regions.Furthermore,we mapped a loss-of-function AGL2 gene using bulked segregant RNA-sequencing assay and proved that AGL2 is essential for seed development.We additionally established an open-access database named MEILAM for easy access to Ds insertional mutations.Overall,our results have provided an efficient workflow for TE insertion identification and rich sequence-indexed mutant resources for maize functional genomic studies.
文摘Three T-DNA insertional embryonic lethal mutants from NASC (The Nottingham Arabidopsis Stock Center)were first checked with their segregation ratio of abortive and normal seeds and the copy number of T-DNA insertion. The N4081 mutant has a segregation ratio of 1:3.04in average and one T-DNA insertion site according to our assay It was therefore chosen for further analysis. To isolate the joint fragment of T-DNA and plan DNA, the plasmid rescue technique waJs used. pEL-7, one of plasmids from left border of T-DNA, which contained pBR322 was selected from ampicillin plate. The T-DNA fragment of pEL-7 was checked by restriction enzyme analysis and Southern Blot. Restriction analysis confirmed the presence of known sites of EcoRI, PstI and PvuII on it.For confirming the presence of flanking plant DNA in this plasmid, pEL-7 DNA was labeled and hybridized with wild type and mutant plant DNA. The Southern Blot indicated the hybridization band in both of them. Furthermore, the junction of T-DNA/plant DNA was subcloned into bluescript SK+ and sequenced by Applied Biosystem 373A Sequencer. The results showed the 822 bp fragment contained a 274 bp sequence, which is 99.6%homolog (273bp/274 bp) to Ti plasmid pTi 15955 DNA.Ten bp of left 25 bp border repeat were also found in the juction of T-DNA and Plant DNA.Taken together, pEL-7 should contain a joint fragment of T-DNA and flanking plant DNA. This plasmid DNA could be used for the isolation of plant gene, which will be helpful to elucidate the relationship between gene function and plant embryo development.
基金supported by the High-Tech R&D Program of China(2006AA10A106)the open funds of the National Key Laboratory of Crop Genetic Improvement and China National Fundamental Fund of Personnel Training (J0730649)
文摘Mutator transposable element （Mu） has been used as an effective tool to clone maize （Zea mays L.） genes. One opaque endosperm mutant （miol6） was identified in a pool of Mu inserted mutants. A modified method, termed the double selected amplification of insertion flanking fragments （DSAIFF）, was employed to isolate the Mu flanking fragments （MFFs） of miol6. The target site duplications （TSDs） isolated from the Msp I and Mse I digested MFFs had a same 9-bp sequence and were confirmed to be the flanking sequence of one identically inserted gene. Co-segregation analysis suggested that the MFFs were associated with the mutant opaque endosperm, and miol6 was mapped in silico onto the physical position ranged from 229 965 021 to 229 965 409 bp of the maize chromosome 4.09 bin. The full-length cDNA of the wild-type gene was obtained by an RT-PCR primer-scanning technique, and Mio16 was found to putatively encode a homolog of the Arabidopsis MAP3K delta-1 protein kinase. RT-PCR result the mRNA expression of miol6 region anchored by primers Mu20 and af276 was not interrupted by Mu insertion. Further researches will be done to elucidate how the expression of miol6 is alternated by Mu insertion.
文摘BACKGROUND Long-term chemotherapy for patients with gastric cancer(GC),facilitated by peripherally inserted central catheter(PICC)catheterization,reduces vascular damage and enhances drug delivery efficiency but carries risks of catheter-related complications.A combination of group psychological nursing and physical mo-vement care significantly mitigates the risk of venous thrombosis and improves psychological well-being,and enhances motor function,underscoring its clinical importance.AIM To assess group psychological and physical movement nursing in preventing venous thrombosis in patients with PICC GC.METHODS Sixty-five GC patients with PICC,admitted from January 2022 to January 2023,were randomly divided into two groups using the lottery method:A control group(n=35,routine nursing)and an observation group(n=30,routine nursing plus psychological nursing and physical movement nursing).Both groups re-ceived continuous care for 2 weeks.Pre-nursing and post-nursing data on psycho-logical state,physical function,chemotherapy-related thrombosis incidence,and cancer-related fatigue were analyzed using SPSS 26.0 and GraphPad Prism 8.0.RESULTS After nursing,both groups showed reduced Hamilton Anxiety Scale scores and increased General Perceived Self-Efficacy Scale scores,with the observation group performing better(P<0.05).The Functional Comprehensive Assessment score for the observation group after nursing was(65.42±2.35)points,lower than the control group’s(62.19±4.33)points(P<0.05).Although no significant difference was observed in the incidence of venous thrombosis between the two groups(χ2=0.815,P=0.367),the observation group had lower incidence.Both groups showed decreased Revised Piper Fatigue Scale scores,with the observation group scoring lower(P<0.05).CONCLUSION Group psychological and physical movement nursing for patients with PICC reduces venous thrombosis risk,improves psychological well-being,cancer-related fatigue,and physical function,making it highly promotable.
文摘An efficient TfOH-catalyzed O—H insertion reaction of α-aryl diazoesters with carboxylic acids is reported.This metal-free protocol provides an operationally simple method for a one-pot assembly of diverse α-acyloxy esters in moderate to high yields with a broad substrate scope.All starting materials are readily available,and the reactions can be conducted in the open air at room temperature.
文摘A blue light-induced formal insertion reaction ofα-siloxy carbene into the C—H bond of 1,3-diketones has been reported.Under the irradiation of blue light,acylsilane converts toα-siloxy carbene,which then undergoes formal C—H bond insertion reaction with the enol form of 1,3-diketone.This method uses readily available and relative stable acylsilane as car-bene precursor,which features a simple and metal-free approach under mild conditions.Moreover,the synthetic potential of this protocol has been demonstrated by performing the reaction on a gram scale with comparable high yield.
文摘There is nonradiative recombination in waveguide region owing to severe carrier leakage,which in turn reduces output power and wall-plug efficiency.In this paper,we designed a novel epitaxial structure,which suppresses carrier leakage by inserting n-Ga_(0.55)In_(0.45)P and p-GaAs_(0.6)P_(0.4) between barriers and waveguide layers,respectively,to modulate the energy band structure and to increase the height of barrier.The results show that the leakage current density reduces by 87.71%,compared to traditional structure.The nonradiative recombination current density of novel structure reduces to 37.411 A/cm^(2),and the output power reaches 12.80 W with wall-plug efficiency of 78.24%at an injection current density 5 A/cm^(2) at room temperature.In addition,the temperature drift coefficient of center wavelength is 0.206 nm/℃at the temperature range from 5℃to 65℃,and the slope of fitted straight line of threshold current with temperature variation is 0.00113.The novel epitaxial structure provides a theoretical basis for achieving high-power laser diode.
文摘Virus-based vectors are widely used in hematopoietic stem cell(HSC)gene therapy,and have the ability to integrate permanently into genomic DNA,thus driving long-term expression of corrective genes in all hematopoietic lineages.To date,HSC gene therapy has been successfully employed in the clinic for improving clinical outcomes in small numbers of patients with X-linked severe combined immunodeficiency(SCID-X1),adenosine deaminase deficiency(ADA-SCID),adrenoleukodystrophy(ALD),thalassemia,chronic granulomatous disease(CGD),and Wiskott-Aldrich syndrome(WAS).However,adverse events were observed during some of these HSC gene therapy clinical trials,linked to insertional activation of proto-oncogenes by integrated proviral vectors leading to clonal expansion and eventual development of leukemia.Numerous studies have been performed to understand the molecular basis of vector-mediated genotoxicity,with the aim of developing safer vectors and lower-risk gene therapy protocols.This review will summarize current information on the mechanisms of insertional mutagenesis in hematopoietic stem and progenitor cells due to integrating gene transfer vectors,discuss the available assays for predicting genotoxicity and mapping vector integration sites,and introduce newlydeveloped approaches for minimizing genotoxicity as a way to further move HSC gene therapy forward into broader clinical application.
基金CAMS Innovation Fund for Medical Sciences(CIFMS)(Nos.2022-I2M-1-013,2022-I2M-1-014,2022-I2M-2-002).
文摘The switchable cross-coupling of indoles and pyridotriazoles through carbene insertion at C_(2)-or C_(3)-positon has been developed in this paper.This highly site-selective C-H carbenoid functionalization is determined by both the Rh-catalyst species and auxiliary groups.[Cp∗RhCl_(2)]_(2) and coordinating pyrimidyl group direct the C-H carbenoid functionalization to occur at the C_(2)-position,while Rh2OAc4 and noncoordinating benzyl group lead the reaction to occur at the C_(3)-position of the indoles.This regioselective C−H functionalization strategy is of significant importance for the discovery of indole drugs.
基金supported by the Korean Institute of Energy Technology Evaluation and Planning(KETEP)grant funded by the Korea Government(MOTIE)(RS-2023-00303581,Multiscale Simulation-Driven Development of Cost-Effective and Stable Aqueous Zn Ion Battery with Energy Density of 110 Wh/L for Energy Storage Systems:A Korea-USA Collaboration)。
文摘Achieving high-performance aqueous zinc-ion batteries requires addressing the challenges associated with the stability of zinc metal anodes,particularly the formation of inhomogeneous zinc dendrites during cycling and unstable surface electrochemistry.This study introduces a practical method for scattering untreated bulk hexagonal boron nitride(h-BN)particles onto the zinc anode surface.During cycling,stabilized zinc fills the interstices of scattered h-BN,resulting in a more favorable(002)orientation.Consequently,zinc dendrite formation is effectively suppressed,leading to improved electrochemical stability.The zinc with scattered h-BN in a symmetric cell configuration maintains stability 10 times longer than the bare zinc symmetric cell,lasting 500 hours.Furthermore,in a full cell configuration withα-MnO_(2) cathode,increased H^(+)ion activity can effectively alter the major redox kinetics of cycling due to the presence of scattered h-BN on the zinc anode.This shift in H^(+)ion activity lowers the overall redox potential,resulting in a discharge capacity retention of 96.1%for 300 cycles at a charge/discharge rate of 0.5 Ag^(-1).This study highlights the crucial role of surface modification,and the innovative use of bulk h-BN provides a practical and effective solution for improving the performance and stability.
基金supported by the grants from the National Key Research and Development Program of China(2022YFD1800604)the China Agriculture Research System(CARS-41)the Heilongjiang Touyan Innovation Team Program,China。
文摘Avian metapneumovirus(aMPV),a paramyxovirus,causes acute respiratory diseases in turkeys and swollen head syndrome in chickens.This study established a reverse genetics system for aMPV subtype B LN16-A strain based on T7 RNA polymerase.Full-length cDNA of the LN16-A strain was constructed by assembling 5 cDNA fragments between the T7 promoter and hepatitis delta virus ribozyme.Transfection of this plasmid,along with the supporting plasmids encoding the N,P,M2-1,and L proteins of LN16-A into BSR-T7/5 cells,resulted in the recovery of aMPV subtype B.To identify an effective insertion site,the enhanced green fluorescent protein(EGFP)gene was inserted into different sites of the LN16-A genome to generate recombinant LN16-As.The results showed that the expression levels of EGFP at the site between the G and L genes of LN16-A were significantly higher than those at the other two sites(between the leader and N genes or replacing the SH gene).To verify the availability of the site between G and L for foreign gene expression,the VP2 gene of very virulent infectious bursal disease virus(vvIBDV)was inserted into this site,and recombinant LN16-A(rLN16A-vvVP2)was successfully rescued.Single immunization of specificpathogen-free chickens with rLN16A-vvVP2 induced high levels of neutralizing antibodies and provided 100%protection against the virulent aMPV subtype B and vvIBDV.Establishing a reverse genetics system here provides an important foundation for understanding aMPV pathogenesis and developing novel vector vaccines.
文摘The ThinKing by Russian watchmaker Konstantin Chaykin is only 1.65 millimeters thick,making it the world's thinnest mechanical wristwatch.The“arms race”for the world's thinnest wristwatch has been in full swing for a few years now,with some of the most prestigious(有声望的)watch brands on Earth trying to one‑up each other with ingenious designs.The latest to claim the coveted title is independent watchmaker Konstantin Chaykin with his ThinKing prototype,a 1.65‑millimeter‑thick masterpiece that is so incredibly thin that it needs a special wristband.The ThinKing's case is so delicate that the simple pressure from being worn is enough to damage it,so the special strap made of alligator leather and elastic(有弹性的)inserts with titanium(钛)has to absorb some of the shocks,twisting and bending.
基金supported by the National Key Research and Development Program Project(No.2021YFB3301300).
文摘Intelligent production is an important development direction in intelligent manufacturing,with intelligent factories playing a crucial role in promoting intelligent production.Flexible job shops,as the main form of intelligent factories,constantly face dynamic disturbances during the production process,including machine failures and urgent orders.This paper discusses the basic models and research methods of job shop scheduling,emphasizing the important role of dynamic job shop scheduling and its response schemes in future research.A multi-objective flexible job shop dynamic scheduling mathematical model is established,highlighting its complex and multi-constraint characteristics under different interferences.A classification discussion is conducted on the dynamic response methods and optimization objectives under machine failures,emergency orders,fuzzy completion times,and mixed dynamic events.The development process of traditional scheduling rules and intelligent methods in dynamic scheduling are also analyzed.Finally,based on the current development status of job shop scheduling and the requirements of intelligent manufacturing,the future development trends of dynamic scheduling in flexible job shops are proposed.
基金financially supported by National Natural Science Foundation of China(No.22207113 to J.Zhang)Guangdong Basic and Applied Basic Research Foundation(No.2021A1515110588to J.Zhang)Natural Science Foundation of Shanghai Municipality(No.22ZR1474000 to L.Wen)。
文摘Uronic acids are prevalent components of crucial glycoconjugates,pivotal in various biological processes.In nature,NDP-uronic acids,the nucleosides-activated uronic acids,serve as glycosylation donors catalyzed by uronosyltransferases(UATs)to construct glycans containing uronic acids.Despite their biological importance,the synthesis of naturally occurring NDP-uronic acids on a large scale remains challenging.Here,we developed an oxidation reaction insertion strategy for the efficient synthesis of NDP-uronic acids,and 11 NDP-uronic acids were successfully prepared in good yield and on a large scale.The prepared NDP-uronic acids can be used to explore new uronosyltransferases and synthesize uronic acids containing carbohydrates for fundamental research.
基金supported by grants(92168103,32171417,2019CXJQ01)from the National Nature Science Foundation of China,Shanghai Municipal GovernmentPeak Disciplines(Type IV)of Institutions of Higher Learning in Shanghai.
文摘Genome rearrangement is an important process that leads to genetic diversity,including mutation-related insertions,deletions,or inversions in the genome[1,2].
基金Supported by the Shandong Province Medical and Health Technology Development Plan,No.202204011069.
文摘BACKGROUND Peripherally inserted central catheters(PICCs)are widely used for administering chemotherapy to breast cancer patients due to their long-term indwelling capability,versatility in drug administration,and flexibility.PICCs infection are a relatively common occurrence,yet there were no reported instances that it can metastasise to the lumbar spine.CASE SUMMARY This case report describes a breast cancer patient who developed a methicillinresistant Staphylococcus aureus lumbar vertebral infection secondary to a PICCrelated infection during chemotherapy.Following PICC removal,bacterial culture confirmed the presence of highly virulent methicillin-resistant Staphylococcus aureus.The patient presented with fever and severe lumbar pain.Lumbar magnetic resonance imaging revealed paraspinal muscle edema from L1 to L3 with abnormal signal intensity in the affected regions,suggestive of vertebral osteomyelitis.Prompt initiation of appropriate antibiotic therapy based on the culture results led to significant improvement in the patient’s lumbar pain.CONCLUSION This case highlights the importance of vigilant infection prevention and control measures to minimize the risk of PICC-related complications,such as bloodstream infections and subsequent metastatic infections.
