To study biomarker of acrylamide （ACR） induced neuropathy, Wistar rats received 20 or 40 mg/kg of ACR by ip injection and the levels of light neurofilament （NF-L）, middle NF （NF-M）, heavy NF （NF-H）, β-actin, ...To study biomarker of acrylamide （ACR） induced neuropathy, Wistar rats received 20 or 40 mg/kg of ACR by ip injection and the levels of light neurofilament （NF-L）, middle NF （NF-M）, heavy NF （NF-H）, β-actin, α-tubulin, and β-tubulin proteins in serum were evaluated using both SDS-PAGE and Western blotting. Compared to controls, NF-L and NF-M decreased,展开更多
The study was conducted to investigate the effect and mechanism of dietary quercetin supplementation on protein utilization of Arbor Acres(AA)broilers.A total of 2401-day-old AA broilers were randomly allocated to fou...The study was conducted to investigate the effect and mechanism of dietary quercetin supplementation on protein utilization of Arbor Acres(AA)broilers.A total of 2401-day-old AA broilers were randomly allocated to four treatments with six replicates,comprising 10 broilers each replicate(60 broilers per treatment).Birds were fed either a corn-soybean meal basal diet without quercetin(control)or a basal diet supplemented with 0.2,0.4 or 0.6 g of quercetin per kg feed,and the trial lasted 42 days.Dietary quercetin supplementation tended to increase the apparent metabolic rate of protein(p=0.076)and the content of serum albumin(p=0.062)in AA broilers.Compared with the control,dietary quercetin supplementation increased the contents of protein in breast muscle(p<0.05)and in thigh muscle(p=0.053).In addition,quercetin up-regulated mRNA expression of insulin-like growth factor 1(IGF-1),phosphatidylinositol 3-kinase(PI3K),target of rapamycin(TOR),ribosomal protein S6 kinase 1(S6K1),eukaryotic translation initiation factor 4E(eIF4E),eukaryotic translation initiation factor 4G(eIF4G),eukaryotic elongation factor 2(eEF2)and eukaryotic translation initiation factor 4B(eIF4B)genes and down-regulated mRNA expression of eukaryotic elongation factor 2 kinase(eEF2K)and eukaryotic initiation factor 4E binding protein1(4E-BP1)genes in breast muscle,thigh muscle and liver of AA broilers(p<0.05).The present results suggested that dietary quercetin supplementation enhanced protein utilization in broilers by activating TOR signaling pathway.展开更多
Rheumatoid arthritis is a chronic multisystem disease of unknown cause. The characteristic feature of RA is persistent inflammatory synovitis. The natural history of disease is such that the early months of the diseas...Rheumatoid arthritis is a chronic multisystem disease of unknown cause. The characteristic feature of RA is persistent inflammatory synovitis. The natural history of disease is such that the early months of the disease are critical period during which reversible joint damage occurs. So early diagnosis of RA and appropriate drug application is the only way to save a patient from this crippling disease. In India, the cost of investigations is a significant factor for most of the patients. Ultrasonography or Power Doppler Ultra Sound (PDUS) has the advantage of being economic in spite of its sensitivity in assessing both inflammatory and destructive changes. The aim of the present study was to evaluate the diagnostic efficiency of PDUS in early rheumatoid arthritis. The study was performed with the patients attending Rheumatology Clinic. A total number of 106 patients of clinically suspected rheumatoid arthritis were studied as per selection criteria. Radiological examinations of hands were done by digital radiography and PDUS in a group of 53 patients, assessment of foot changes by PDUS and Digital Radiography were done in another similar group of 53 patients. Final diagnosis by ACR EULAR-2010 criteria is done for all the patients. The comparative study reveals that synovial vascularity as demonstrated by PDUS is much more effective in diagnosing early rheumatoid arthritis, both in hand and in feet than digital radiograph. PDUS of feet may yield earlier and better findings than hands, which is conventionally used in patients suffering from early rheumatoid arthritis.展开更多
In the ongoing arms race between bacteria and bacteriophages,bacteriophages have evolved anti-CRISPR proteins to counteract bacterial CRISPR-Cas systems.Recently,AcrⅡA25.1 and AcrⅡA32 have been found to effectively ...In the ongoing arms race between bacteria and bacteriophages,bacteriophages have evolved anti-CRISPR proteins to counteract bacterial CRISPR-Cas systems.Recently,AcrⅡA25.1 and AcrⅡA32 have been found to effectively inhibit the activity of Spy Cas9 both in bacterial and human cells.However,their molecular mechanisms remain elusive.Here,we report the cryo-electron microscopy structures of ternary complexes formed by AcrⅡA25.1 and AcrⅡA32 bound to Spy Cas9-sg RNA.Using structural analysis and biochemical experiments,we revealed that AcrⅡA25.1 and AcrⅡA32 recognize a novel,previously-unidentified anti-CRISPR binding site on Spy Cas9.We found that both AcrⅡA25.1 and AcrⅡA32 directly interact with the WED domain,where they spatially obstruct conformational changes of the WED and PI domains,thereby inhibiting Spy Cas9 from recognizing protospacer adjacent motif(PAM)and unwinding double-stranded DNA.In addition,they may inhibit nuclease activity by blocking the dynamic conformational changes of the Spy Cas9 surveillance complex.In summary,our data elucidate the inhibition mechanisms of two new anti-CRISPR proteins,provide new strategies for the modulation of Spy Cas9 activity,and expand our understanding of the diversity of anti-CRISPR protein inhibition mechanisms.展开更多
Gluconobacter oxydans is an important Gram-negative industrial microorganism that produces vitamin C and other products due to its efficient membrane-bound dehydrogenase system.Its incomplete oxidation system has many...Gluconobacter oxydans is an important Gram-negative industrial microorganism that produces vitamin C and other products due to its efficient membrane-bound dehydrogenase system.Its incomplete oxidation system has many crucial industrial applications.However,it also leads to slow growth and low biomass,requiring further metabolic modification for balancing the cell growth and incomplete oxidation process.As a non-model strain,G.oxydans lacks efficient genome editing tools and cannot perform rapid multi-gene editing and complex metabolic network regulation.In the last 15 years,our laboratory attempted to deploy multiple CRISPR/Cas systems in different G.oxydans strains and found none of them as functional.In this study,Cpf1-based or dCpf1-based CRISPRi was constructed to explore the targeted binding ability of Cpf1,while Cpf1–FokI was deployed to study its nuclease activity.A study on Cpf1 found that the CRISPR/Cpf1 system could locate the target genes in G.oxydans but lacked the nuclease cleavage activity.Therefore,the CRISPR/Cpf1–FokI system based on FokI nuclease was constructed.Single-gene knockout with efficiency up to 100%and double-gene iterative editing were achieved in G.oxydans.Using this system,AcrVA6,the anti-CRISPR protein of G.oxydans was discovered for the first time,and efficient genome editing was realized.展开更多
基金supported by the National Natural Science Fund of China (No.30872088)Technology Development Plan of Shandong Province(No.2012GSF11854)
文摘To study biomarker of acrylamide （ACR） induced neuropathy, Wistar rats received 20 or 40 mg/kg of ACR by ip injection and the levels of light neurofilament （NF-L）, middle NF （NF-M）, heavy NF （NF-H）, β-actin, α-tubulin, and β-tubulin proteins in serum were evaluated using both SDS-PAGE and Western blotting. Compared to controls, NF-L and NF-M decreased,
基金Supported by the National Natural Science Foundation of China(31872377)。
文摘The study was conducted to investigate the effect and mechanism of dietary quercetin supplementation on protein utilization of Arbor Acres(AA)broilers.A total of 2401-day-old AA broilers were randomly allocated to four treatments with six replicates,comprising 10 broilers each replicate(60 broilers per treatment).Birds were fed either a corn-soybean meal basal diet without quercetin(control)or a basal diet supplemented with 0.2,0.4 or 0.6 g of quercetin per kg feed,and the trial lasted 42 days.Dietary quercetin supplementation tended to increase the apparent metabolic rate of protein(p=0.076)and the content of serum albumin(p=0.062)in AA broilers.Compared with the control,dietary quercetin supplementation increased the contents of protein in breast muscle(p<0.05)and in thigh muscle(p=0.053).In addition,quercetin up-regulated mRNA expression of insulin-like growth factor 1(IGF-1),phosphatidylinositol 3-kinase(PI3K),target of rapamycin(TOR),ribosomal protein S6 kinase 1(S6K1),eukaryotic translation initiation factor 4E(eIF4E),eukaryotic translation initiation factor 4G(eIF4G),eukaryotic elongation factor 2(eEF2)and eukaryotic translation initiation factor 4B(eIF4B)genes and down-regulated mRNA expression of eukaryotic elongation factor 2 kinase(eEF2K)and eukaryotic initiation factor 4E binding protein1(4E-BP1)genes in breast muscle,thigh muscle and liver of AA broilers(p<0.05).The present results suggested that dietary quercetin supplementation enhanced protein utilization in broilers by activating TOR signaling pathway.
文摘Rheumatoid arthritis is a chronic multisystem disease of unknown cause. The characteristic feature of RA is persistent inflammatory synovitis. The natural history of disease is such that the early months of the disease are critical period during which reversible joint damage occurs. So early diagnosis of RA and appropriate drug application is the only way to save a patient from this crippling disease. In India, the cost of investigations is a significant factor for most of the patients. Ultrasonography or Power Doppler Ultra Sound (PDUS) has the advantage of being economic in spite of its sensitivity in assessing both inflammatory and destructive changes. The aim of the present study was to evaluate the diagnostic efficiency of PDUS in early rheumatoid arthritis. The study was performed with the patients attending Rheumatology Clinic. A total number of 106 patients of clinically suspected rheumatoid arthritis were studied as per selection criteria. Radiological examinations of hands were done by digital radiography and PDUS in a group of 53 patients, assessment of foot changes by PDUS and Digital Radiography were done in another similar group of 53 patients. Final diagnosis by ACR EULAR-2010 criteria is done for all the patients. The comparative study reveals that synovial vascularity as demonstrated by PDUS is much more effective in diagnosing early rheumatoid arthritis, both in hand and in feet than digital radiograph. PDUS of feet may yield earlier and better findings than hands, which is conventionally used in patients suffering from early rheumatoid arthritis.
基金supported by the National Key Research and Development Program of China(2023YFF1000200)the National Natural Science Foundation of China(U21A20276)+1 种基金the Tencent Foundation through the XPLORER PRIZE and the New Cornerstone Science Foundation to Z.H.,Heilongjiang Touyan Team(HITTY-20190034 to Z.H.)Natural Science Foundation of Heilongjiang Province of China(YQ2023C032 to Y.Z.)。
文摘In the ongoing arms race between bacteria and bacteriophages,bacteriophages have evolved anti-CRISPR proteins to counteract bacterial CRISPR-Cas systems.Recently,AcrⅡA25.1 and AcrⅡA32 have been found to effectively inhibit the activity of Spy Cas9 both in bacterial and human cells.However,their molecular mechanisms remain elusive.Here,we report the cryo-electron microscopy structures of ternary complexes formed by AcrⅡA25.1 and AcrⅡA32 bound to Spy Cas9-sg RNA.Using structural analysis and biochemical experiments,we revealed that AcrⅡA25.1 and AcrⅡA32 recognize a novel,previously-unidentified anti-CRISPR binding site on Spy Cas9.We found that both AcrⅡA25.1 and AcrⅡA32 directly interact with the WED domain,where they spatially obstruct conformational changes of the WED and PI domains,thereby inhibiting Spy Cas9 from recognizing protospacer adjacent motif(PAM)and unwinding double-stranded DNA.In addition,they may inhibit nuclease activity by blocking the dynamic conformational changes of the Spy Cas9 surveillance complex.In summary,our data elucidate the inhibition mechanisms of two new anti-CRISPR proteins,provide new strategies for the modulation of Spy Cas9 activity,and expand our understanding of the diversity of anti-CRISPR protein inhibition mechanisms.
基金This work was supported by the National Natural Science Foundation of China(Key Program,31830068),China National Postdoctoral Program for Innovative Talents(BX20230147),China Postdoctoral Science Foundation(2023M741406),Jiangsu Funding Program for Excellent Postdoctoral Talent(2023ZB631).
文摘Gluconobacter oxydans is an important Gram-negative industrial microorganism that produces vitamin C and other products due to its efficient membrane-bound dehydrogenase system.Its incomplete oxidation system has many crucial industrial applications.However,it also leads to slow growth and low biomass,requiring further metabolic modification for balancing the cell growth and incomplete oxidation process.As a non-model strain,G.oxydans lacks efficient genome editing tools and cannot perform rapid multi-gene editing and complex metabolic network regulation.In the last 15 years,our laboratory attempted to deploy multiple CRISPR/Cas systems in different G.oxydans strains and found none of them as functional.In this study,Cpf1-based or dCpf1-based CRISPRi was constructed to explore the targeted binding ability of Cpf1,while Cpf1–FokI was deployed to study its nuclease activity.A study on Cpf1 found that the CRISPR/Cpf1 system could locate the target genes in G.oxydans but lacked the nuclease cleavage activity.Therefore,the CRISPR/Cpf1–FokI system based on FokI nuclease was constructed.Single-gene knockout with efficiency up to 100%and double-gene iterative editing were achieved in G.oxydans.Using this system,AcrVA6,the anti-CRISPR protein of G.oxydans was discovered for the first time,and efficient genome editing was realized.
