A series of new O6-BG derivatives （14-21,23-30） were synthesized as inactivators of O6-Alkylguanine-DNA alkyltransferase （AGT）, and their ability to inhibit AGT was preliminary evaluated by MTT with O6-BG as the c...A series of new O6-BG derivatives （14-21,23-30） were synthesized as inactivators of O6-Alkylguanine-DNA alkyltransferase （AGT）, and their ability to inhibit AGT was preliminary evaluated by MTT with O6-BG as the control. The result suggested compound 30 displayed a higher activity than O6-BG.展开更多
The removal of ammonia nitrogen(NH_(4)^(+)-N)and bacteria from aquaculture wastewater holds paramount ecological and production significance.In this study,Pt/RuO_(2)/g-C_(3)N_(4)photocatalysts were prepared by deposit...The removal of ammonia nitrogen(NH_(4)^(+)-N)and bacteria from aquaculture wastewater holds paramount ecological and production significance.In this study,Pt/RuO_(2)/g-C_(3)N_(4)photocatalysts were prepared by depositing Pt and RuO_(2)particles onto g-C_(3)N_(4).The physicochemical properties of photocatalysts were explored by X-ray photoelectron spectroscopy(XPS),scanning electron microscopy(SEM),X-ray diffraction(XRD),and UV–vis diffuse reflectance spectrometer(UV–vis DRS).The photocatalysts were then applied to the removal of both NH_(4)^(+)-N and bacteria from simulated mariculture wastewater.The results clarified that the removals of both NH_(4)^(+)-N and bacteria were in the sequence of g-C_(3)N_(4)<RuO_(2)/g-C_(3)N_(4)<Pt/g-C_(3)N_(4)<Pt/RuO_(2)/g-C_(3)N_(4).This magnificent photocatalytic ability of Pt/RuO_(2)/g-C_(3)N_(4)can be interpreted by the transfer of holes from g-C_(3)N_(4)to RuO_(2)to facilitate the in situ generation of HClO from Cl^(−)in wastewater,while Pt extracts photogenerated electrons for H_(2)formation to enhance the reaction.The removal of NH_(4)^(+)-N and disinfection effect were more pronounced in simulated seawater than in purewater.The removal efficiency ofNH_(4)^(+)-N increases with an increase in pH of wastewater,while the bactericidal effect was more significant under a lower pH in a pH range of 6–9.In actual seawater aquaculture wastewater,Pt/RuO_(2)/g-C_(3)N_(4)still exhibits effective removal efficiency of NH_(4)^(+)-N and bactericidal performance under sunlight.This study provides an alternative avenue for removement of NH_(4)^(+)-N and bacteria from saline waters under sunlight.展开更多
Solar-driven Fenton-like reactions are promising strategies for degrading pharmaceutical wastewater to address environmental challenges and antibiotic pollution.However,its efficacy is limited by suboptimal light abso...Solar-driven Fenton-like reactions are promising strategies for degrading pharmaceutical wastewater to address environmental challenges and antibiotic pollution.However,its efficacy is limited by suboptimal light absorption efficiency,rapid charge recombination,and inadequate interfacial charge transfer.In this study,an inorganic/organic S-scheme photo-Fenton system of pseudobrookite/carbon nitride(FTOCN)was synthesized via a hydrothermally coupled calcination process for the effective purification of tetracycline antibiotics under visible-light irradiation.The optimized FTOCN-2 heterostructure exhibits a significantly enhanced TC degradation capacity of 90%within 60 min.The rate constant of FTOCN-2 is 1.6 and 5.2 times greater than those of FTO and CN,respectively.Furthermore,FTOCN exhibits high antibacterial efficacy,highlighting its potential application in the purification of natural water.Measurements via a range of analytical techniques,including Kelvin probe force microscopy,density functional theory calculations,in situ X-ray photoelectron spectroscopy,and femtosecond transient absorption spectroscopy,corroborate the S-scheme mechanism.This study provides a novel perspective for the development of photo-Fenton systems with S-scheme heterojunctions for water purification.展开更多
Minks are highly susceptible to SARS-CoV-2,and have transmitted SARS-CoV-2 to humans.Oral immunization is one of the most promising strategies to prevent SARS-CoV-2 infection and transmission in minks.Here,we generate...Minks are highly susceptible to SARS-CoV-2,and have transmitted SARS-CoV-2 to humans.Oral immunization is one of the most promising strategies to prevent SARS-CoV-2 infection and transmission in minks.Here,we generated 3 recombinant rabies viruses(RABV),rERAG_(333E)/S6P,rERAG_(333E)/DS6P and rERAG_(333E)/BA2S6P,expressing the prefusion-stabilized SARS-CoV-2 spike protein of wild-type(S6P),δ(DS6P)or BA.2(BA2S6P)strain based on an oral rabies vaccine candidate(rERAG_(333E)).Oral or inactivated immunization of the 3 RABVs monovalent or trivalent were safe,and induced robust RABV neutralizing antibody and cross-antibody responses against the three SARS-CoV-2 in mice and minks.The challenge tests showed that 2 doses of rERAG_(333E)-S6P as an oral or inactivated vaccine completely protected mice against mouse-adapted SARS-CoV-2 infection in the upper and lower respiratory tracts,and largely prevented viral replication and lung damage caused by wild-type SARS-CoV-2infection in minks.Notably,we also confirmed that 2 doses of rERAG_(333E)-S6P as an oral or inactivated vaccine can largely protect minks against wild-type SARS-CoV-2 transmission via respiratory droplets.Our findings suggest that rERAG_(333E)-based COVID-19 vaccines appear to be suitable oral candidates to protect minks from SARS-CoV-2infection and transmission,and may serve as inactivated vaccines for further investigation in humans.展开更多
Dent disease is a rare X-linked recessively inherited renal tubulopathy,caused by variants in the CLCN5(Dent disease type 1,DD1)and OCRL(Dent disease type 2,DD2)genes,and characterized by low molecular weight proteinu...Dent disease is a rare X-linked recessively inherited renal tubulopathy,caused by variants in the CLCN5(Dent disease type 1,DD1)and OCRL(Dent disease type 2,DD2)genes,and characterized by low molecular weight proteinuria,hypercalciuria,microscopic hematuria,or nephrocalcinosis.In the current study,we collected and analyzed clinical data and genetic testing results of 21 children diagnosed with Dent disease,who were hospitalized at the Department of Nephrology,Children's Hospital of Nanjing Medical University between January 2014 and August 2023,aiming to assist in the early diagnosis and treatment of these patients.Among the 21 patients,16(76.19%)had CLCN5 variants,and five(23.81%)had OCRL variants,and four of the variants were novel.All patients presented with low molecular weight proteinuria,14(66.67%)of whom had nephroticrange proteinuria.Eight patients underwent renal biopsies because of diagnostic uncertainty.We transfected the novel CLCN5 missense variant(p.G222R)and OCRL missense variant(p.I371T)plasmids into both HEK293 and HK-2 cells and found a significantly lower expression of the OCRL1 protein in the transfected cells than in the wild-type cells(P<0.05).Moreover,we observed an extremely skewed X-chromosome inactivation pattern in a female patient carrying the same novel CLCN5 variant,as assessed by the human androgen receptor gene assay.These findings provide insight into the clinical characteristics of Dent disease in Chinese patients and may shed light on its pathogenesis.展开更多
Ionizing radiation presents an important solution for virus inactivation.However,its efficacy for severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)inactivation and the underlying mechanisms remain unclear.Th...Ionizing radiation presents an important solution for virus inactivation.However,its efficacy for severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)inactivation and the underlying mechanisms remain unclear.This study demonstrates radiosensitivity and radiation-induced biological changes in SARS-CoV-2 using 20 wild-type and mutant strains.The results show that 1.2 kGy of electron beam(E-beam)or 0.9 kGy of X-ray irradiation can eliminate 99.99%of SARS-CoV-2 particles.The Delta and various Omicron variants exhibit heightened sensitivity to radiation compared to the wild-type,showing nearly 99.99%inactivation efficiency at 1.0 and 0.8 kGy.The relationship between irradiation dose and the logarithmic reduction in virus load adheres to a dose-response model,characterized by extremely narrow windows.Spike(S)protein disruption,rather than the commonly accepted nucleic acid cleavage,is identified as the primary inactivation mechanism(triggering a conformation transition of S protein from pre-fusion to post-fusion with minimal impact on nucleic acid integrity).This study introduces the concept of targeting critical proteins in coronavirus inactivation,offering valuable insight for infectious coronavirus disease control and vaccine development.展开更多
The cranial base synchondroses,comprised of opposite-facing bidirectional chondrocyte layers,drive anteroposterior cranial base growth.In humans,RUNX2 haploinsufficiency causes cleidocranial dysplasia associated with ...The cranial base synchondroses,comprised of opposite-facing bidirectional chondrocyte layers,drive anteroposterior cranial base growth.In humans,RUNX2 haploinsufficiency causes cleidocranial dysplasia associated with deficient midfacial growth.However,how RUNX2 regulates chondrocytes in the cranial base synchondroses remains unknown.To address this,we inactivated Runx2 in postnatal synchondrosis chondrocytes using a tamoxifen-inducible Fgfr3-creER(Fgfr3-Runx2cKO)mouse model.Fgfr3-Runx2cKO mice displayed skeletal dwarfism and reduced anteroposterior cranial base growth associated with premature synchondrosis ossification due to impaired chondrocyte proliferation,accelerated hypertrophy,apoptosis,and osteoclast-mediated cartilage resorption.Lineage tracing reveals that Runx2-deficient Fgfr3+cells failed to differentiate into osteoblasts.Notably,Runx2-deficient chondrocytes showed an elevated level of FGFR3 and its downstream signaling components,pERK1/2 and SOX9,suggesting that RUNX2 downregulates FGFR3 in the synchondrosis.This study unveils a new role of Runx2 in cranial base chondrocytes,identifying a possible RUNX2-FGFR3-MAPK-SOX9 signaling axis that may control cranial base growth.展开更多
Dear Editor,Viruses of the genus Orthoebolavirus cause sporadic outbreaks of severe haemorrhagic fever,with case fatality rates ranging from 25%to 90%(Mahanty and Bray,2004).Six species of the virus(Orthoebolavirus za...Dear Editor,Viruses of the genus Orthoebolavirus cause sporadic outbreaks of severe haemorrhagic fever,with case fatality rates ranging from 25%to 90%(Mahanty and Bray,2004).Six species of the virus(Orthoebolavirus zairense,sudanense,bundibugyoense,taiense,restonense,and bombaliense)have so far been identified(Biedenkopf et al.,2023).Among these,Orthoebolavirus zairense,commonly known as Ebola virus(EBOV),stands out as the most virulent.Given its high contagiousness and lethality,EBOV must be manipulated under biosafety level 4(BSL-4)conditions,as stipulated by the the People's Republic of China's list of human pathogenic microorganisms(National Health Commission of the People’s Republic of China,2023).Prior to being removed from a BSL-4 laboratory,it is imperative that infectious EBOV undergoes complete inactivation.Here we systematically evaluate viral thermostability under BSL-4 containment conditions,demonstrating EBOV's marked thermotolerance.展开更多
The natural curcumin-mediated photodynamic inactivation(PDI)was developed,and its inactivation potency against Fusarium graminearum in vitro and in vivo was systematically investigated by fluorescence probe assay,tryp...The natural curcumin-mediated photodynamic inactivation(PDI)was developed,and its inactivation potency against Fusarium graminearum in vitro and in vivo was systematically investigated by fluorescence probe assay,trypan blue staining,scanning electron microscope(SEM),confocal laser scanning microscopy(CLSM),etc.Results showed that under the irradiation of blue LED,the photosensitizer of curcumin was excited to generate massive reactive oxygen species(ROS)in the cells of F.graminearum,and the PDI completely inactivated their mycelia and spores under the treatment of 150μM curcumin and 10.8 J/cm^(2)irradiation.Further analysis found that the PDI ruptured the cellular microstructures,damaged the cell membrane by increasing its permeability and oxidizing the lipids,degraded the intracellular DNA and proteins inside the spores of F.graminearum.Meanwhile,the PDI also potently killed>99.99%spores of F.graminearum on maize under the treatment of 200μM curcumin and 10.8 J/cm^(2)irradiation.Moreover,the PDI suppressed the production of zearalenone(ZEN),and residual ZEN could not be detected after the storage of maize for 10 days.Therefore,this study systematically explored the inactivation efficiency of curcumin-mediated PDI against both the mycelia and spores of F.graminearum,which provides a valid and promising method to control the fungal hazards in grains.展开更多
Porcine epidemic diarrhea virus(PEDV)infection causes acute watery diarrhea in neonatal piglets,leading to substantial economic losses within the pig farming industry.This study demonstrates that clofazimine(CFZ)signi...Porcine epidemic diarrhea virus(PEDV)infection causes acute watery diarrhea in neonatal piglets,leading to substantial economic losses within the pig farming industry.This study demonstrates that clofazimine(CFZ)significantly inhibits PEDV replication in a dose-dependent manner in vitro,with negligible cytotoxicity.Findings from our time-of-addition assays indicate that CFZ effectively disrupts multiple stages of the viral infection cycle.Using a CoV-RdRp-Gluc reporter system,we evaluated the potency of CFZ against PEDV RNA-dependent RNA polymerase(RdRp),and determined a low IC50 value of 0.1364μM.Molecular docking studies further confirmed that CFZ has high binding affinity at the active sites of the spike protein and RdRp protein in PEDV.Transcriptome analysis of Vero E6 cells,with and without CFZ treatment,revealed a significant change in transcriptional activity at 8 h postinfection(hpi).Moreover,the simultaneous application of CFZ and nucleoside analogs showed enhanced the anti-PEDV effect of CFZ in vitro.Our study underscores the potential of CFZ as a viable therapeutic agent against PEDV.展开更多
BACKGROUND Autoimmune encephalitis(AE)is a rare and recently described neuroinflammatory disease associated with specific autoantibodies.Anti-leucine-rich glioma inactivated 1(anti-LGI1)encephalitis is a rare but trea...BACKGROUND Autoimmune encephalitis(AE)is a rare and recently described neuroinflammatory disease associated with specific autoantibodies.Anti-leucine-rich glioma inactivated 1(anti-LGI1)encephalitis is a rare but treatable type of AE discovered in recent years.Alzheimer’s disease(AD)is a degenerative brain disease and the most common cause of dementia.AD may undergo a series of pathological physiological changes in brain tissue 20 years before the onset of typical symptoms.The stage of mild cognitive impairment(MCI)that occurs during this process,known as MCI due to AD,is the earliest stage with clinical symptoms.MCI is typically categorized into two subtypes:Amnestic MCI(aMCI)and non-aMCI.CASE SUMMARY This report describes a patient with rapid cognitive impairment,diagnosed with anti-LGI1 antibody-mediated AE and aMCI,and treated at Peking University Shenzhen Hospital in March 2023.The patient was hospitalized with acute memory decline for more than 3 months.Both the cerebrospinal fluid and serum were positive for anti-LGI1 antibodies,biomarkers of AD coexisting in the patient’s cerebrospinal fluid.Following combination treatment with immunoglobulin therapy and glucocorticoid,plus inhibition of acetylcholinesterase,the patient’s cognitive function significantly improved.Throughout the 3-month follow-up period,a sustained improvement in cognitive function was observed.The results of serum anti-LGI1 antibody were negative.CONCLUSION This case has raised awareness of the possible interaction between AE and early AD(including MCI due to AD),and alerted clinicians to the possibility of concurrent rare and common diseases in patients presenting with cognitive impairment.展开更多
Membrane-initiated estrogen receptorα(mERα)signaling has been shown to affect bone mass in murine models.However,it remains unknown which cell types mediate the mERα-dependent effects on bone.In this study,we gener...Membrane-initiated estrogen receptorα(mERα)signaling has been shown to affect bone mass in murine models.However,it remains unknown which cell types mediate the mERα-dependent effects on bone.In this study,we generated a novel mouse model with a conditional C451A mutation in Esr1,which enables selective knockout of the palmitoylation site essential for the membrane localization of ERα(C451A^(f/f)).First,we used Runx2-Cre mice to generate Runx2-C451A^(f/f)mice with conditional inactivation of mERαsignaling in Runx2-expressing osteoblast lineage cells.No significant changes were observed in body weight,weights of estrogen-responsive organs,or serum concentrations of estradiol between female Runx2-C451A^(f/f)and homozygous C451A^(f/f)littermate controls.High-resolution microcomputed tomography analysis showed a consistent decrease in cortical bone mass in the tibia,femur,and vertebra L5 of Runx2-C451A^(f/f)mice and three-point bending analysis of humerus revealed an impaired mechanical bone strength in Runx2-C451A^(f/f)female mice compared to controls.Additionally,primary osteoblast cultures from mice lacking mERαsignaling showed impaired differentiation compared to controls.展开更多
H_(2)O_(2)is an environmentally friendly oxidizing agent with minimal secondary pollution;however,its application has always been constrained by factors such as storage and transportation.In this study,we propose an i...H_(2)O_(2)is an environmentally friendly oxidizing agent with minimal secondary pollution;however,its application has always been constrained by factors such as storage and transportation.In this study,we propose an innovative method for storing and releasing H_(2)O_(2)using hydrogels.Commercial hydrogels(sodium polyacrylate)can undergo swelling and absorb H_(2)O_(2)in aqueous solutions,and the swollen hydrogel can continuously release H_(2)O_(2)under osmotic pressure.And the characteristics of osmotic pressure drive ensure the recyclability of hydrogel for H_(2)O_(2)storage.Experimental results demonstrate that H_(2)O_(2)can stably exist within the hydrogel for an extended period,and this strategy helps to avoid explosion the risk and potential environmental hazards during the transportation of H_(2)O_(2).Finally,experiments confirm that the hydrogel controlled sustained release of H_(2)O_(2)is effective in both Fenton reactions and the process of bacterial inactivation.This work introduces new ideas for the storage of H_(2)O_(2),and the sustained release of H_(2)O_(2)may have significant implications in the fields of healthcare,environmental science,catalysis,and beyond.展开更多
Coronaviruses are single-stranded,positive-sense RNA enveloped viruses that have posed a significant threat to human health over the past few decades,particularly severe acute respiratory syndrome coronavirus(SARS-CoV...Coronaviruses are single-stranded,positive-sense RNA enveloped viruses that have posed a significant threat to human health over the past few decades,particularly severe acute respiratory syndrome coronavirus(SARS-CoV),Middle East respiratory syndrome coronavirus(MERS-CoV),and SARS-CoV-2.These viruses have caused widespread infections and fatalities,with profound impacts on global economies,social life,and public health systems.Due to their broad host range in natural settings and the consequent high potential for zoonotic spillover events,a thorough investigation of the common viral mechanisms and the identification of druggable targets for pan-coronavirus antiviral development are of utmost importance.展开更多
ZnO/Znml2O4 nanocomposites with heteronanostructures were successfully prepared by co-precipitation method. The as-prepared samples were characterized by HRTEM, TEM, XRD, BET, TG-DTA, and UV-Vis spectra techniques. Th...ZnO/Znml2O4 nanocomposites with heteronanostructures were successfully prepared by co-precipitation method. The as-prepared samples were characterized by HRTEM, TEM, XRD, BET, TG-DTA, and UV-Vis spectra techniques. The photoeatalytic activities of the as-prepared samples were evaluated by the photocatalytic degradation of methyl orange and inactivation of Escherichia coli in suspension under the irradiation of the simulated sunlight. The effects of compositions, calcination temperatures, concentration ofphotocatalysts and light source on the photocatalytic activities were systematically studied. The results show that when the concentration of ZnO/ZnA1204 photocatalyst with the starting Zn to Al molar ratio of 1：1.5 calcined at 600 ℃ is 1.0 g/L, the maximum photocatalytic degradation rate of 98.5% can be obtained in 50 min under the irradiation of the simulated sunlight. Under the same conditions, an inactivation rate of 99.8% for E.coli is achieved in 60 min.展开更多
β-N-Acetyl-D-glucosaminidase （NAGase, EC.3.2.1.52） is chitinolytic enzymes and disintegrate dimmer and trimer a composition of oligomers of N-acetyl-β-D-glucosamine （NAG） into monomer. Prawn （P. vannamei） NAG...β-N-Acetyl-D-glucosaminidase （NAGase, EC.3.2.1.52） is chitinolytic enzymes and disintegrate dimmer and trimer a composition of oligomers of N-acetyl-β-D-glucosamine （NAG） into monomer. Prawn （P. vannamei） NAGase is involved in digestion and molting processes. Some pollutants in seawater affect the enzyme activity causing loss of the biological function of the enzyme, which affects the exuviating shell and threatens the survival of the animal. The effect of formaldehyde on prawn （P. vannamei） β-N-acetyl-D-glucosaminidase activity for the hydrolysis of pNP-NAG has been studied. The results show that formaldehyde, at appropriate concentrations, can lead to reversible inactivation of the enzyme, and the IC50 is estimated to be 1.05mol· L^-1. The inactivation mechanism obtained from Lineweaver-Burk plots shows that the inactivation of the enzyme by formaldehyde belongs to the competitive type. The inactivation kinetics of the enzyme by formaldehyde has been studied using the progress-of-substrate-reaction method described by Tsou, and the rate constants have been determined. The results show that k＋0 is much larger than k-0, indicating the free enzyme molecule is fragile in the formaldehyde solution.展开更多
AIM: To develop and optimize cDNA representational difference analysis (cDNA RDA) method and to identify and clone garlic up-regulated genes in human gastric cancer (HGC) cells. METHODS: We performed cDNA RDA method b...AIM: To develop and optimize cDNA representational difference analysis (cDNA RDA) method and to identify and clone garlic up-regulated genes in human gastric cancer (HGC) cells. METHODS: We performed cDNA RDA method by using abundant double-stranded cDNA messages provided by two self-constructed cDNA libraries (Allitridi-treated and paternal HGC cell line BGC823 cells cDNA libraries respectively). Bam H I and Xho I restriction sites harbored in the library vector were used to select representations. Northern and Slot blots analyses were employed to identify the obtained difference products. RESULTS: Fragments released from the cDNA library vector after restriction endonuclease digestion acted as good marker indicating the appropriate digestion degree for library DNA. Two novel expressed sequence tags (ESTs) and a recombinant gene were obtained. Slot blots result showed a 8-fold increase of glia-derived nexin/protease nexin 1 (GDN/PN1) gene expression level and 4-fold increase of hepatitis B virus x-interacting protein (XIP) mRNA level in BGC823 cells after Allitridi treatment for 72h. CONCLUSION: Elevated levels of GDN/PN1 and XIP mRNAs induced by Allitridi provide valuable molecular evidence for elucidating the garlic's efficacies against neurodegenerative and inflammatory diseases. Isolation of a recombinant gene and two novel ESTs further show cDNA RDA based on cDNA libraries to be a powerful method with high specificity and reproducibility in cloning differentially expressed genes.展开更多
By 30% - 60% (NH4)(2)SO4 fractional precipitation, anion-exchange chromatography on DEAE-Sepharose CL-6B, gel filtration on Sephacryl S-200 and anion-exchange chromatography on Waters AP-1 column (Protein(PM)-Pak DEAE...By 30% - 60% (NH4)(2)SO4 fractional precipitation, anion-exchange chromatography on DEAE-Sepharose CL-6B, gel filtration on Sephacryl S-200 and anion-exchange chromatography on Waters AP-1 column (Protein(PM)-Pak DEAE 15HR), a proteinase which can inactivate soybean trypsin inhibitor (STI) was purified from mung bean ( Vigna rabiata (L.) Wilezek) sprouts. Its molecular weight was estimated to be 29.8 kD by SDS-PAGE, and its K-m and V-max for STI were 769.2 N-alpha -benzoyl-L-arginine ethyl ester BAEE/mL and 115.3 BAEE . mL(-1) . min(-1) respectively. This proteinase was stable at temperatures lower than 50 degreesC and pH 6.5 - 8.5, and 90.91% STI activity of defatted soybean powder was inactivated by this preparation, with proteolytic activity 5 000 BAEE/mL at 50 degreesC and pH 8.0 in 4 h.展开更多
OBJECTIVE: To investigate the effect of Danggui Buxue Tang(DBT), a decoction from Traditional Chinese Medicine, on bleomycin-induced pulmonary fibrosis in rats, and to propose the possible underlying mechanism.METHODS...OBJECTIVE: To investigate the effect of Danggui Buxue Tang(DBT), a decoction from Traditional Chinese Medicine, on bleomycin-induced pulmonary fibrosis in rats, and to propose the possible underlying mechanism.METHODS: Forty male Sprague-Dawley rats were randomly divided into sham group, model group,prednisone group and DBT group. Pulmonary fibrosis rat model was established by intratracheal injection with bleomycin. Body weight and lung index were monitored. Histopathologic examination and collagen deposition were determined using Hematoxylin and eosin(HE) and Masson's trichrome staining. Immunohistochemistry staining was applied to observe the expression of alpha-smooth muscle actin(α-SMA). m RNA expression of α-SMA,collagen Ⅰ and collagen Ⅲ were measured by realtime fluorescence quantitative PCR(RT-q PCR). Inflammatory cytokines, including tumor necrosis factor alpha(TNF-α), interleukin-6(IL-6) and IL-1β in serum were detected by Enzyme-linked immunosorbent assay. Alkali hydrolysis method was conducted to investigate the content of hydroxyproline(HYP). Transforming growth factor-β1(TGF-β1),Smad3 and plasminogen activator inhibitor-1(PAI-1) protein level were examined by Western blot assay.RESULTS: DBT significantly reduced the severity of bleomycin-induced pulmonary fibrosis and inflammation as indicated by minimizing the lost of weight, and by lowering the levels of lung index, inflammation score, Ashcroft score, collagen volume fraction(%), HYP, α-SMA, collagen Ⅰ, collagen Ⅲ,TNF-α, IL-6, IL-1β, TGF-β1, Smad3 and PAI-1, consistent with the effect of prednisone.CONCLUSION: Our findings suggest that DBT is able to ameliorate the pulmonary fibrosis, the possible mechanism may involve inhibition of pulmonary inflammation and collagen deposition, possibly via suppressing TGF-β1/Smad3/PAI-1 signaling pathway.展开更多
A multivalent inactivated Escherichia coli vaccine for forest musk deer by using serotypes O4,O26,and O139 with Al（OH）3 adjuvant was prepared.The vaccine did not cause any adverse reactions in forest musk deer.The i...A multivalent inactivated Escherichia coli vaccine for forest musk deer by using serotypes O4,O26,and O139 with Al（OH）3 adjuvant was prepared.The vaccine did not cause any adverse reactions in forest musk deer.The immunogenic effects of the vaccine were experimentally investigated in pregnant and young forest musk deer.The serum antibody titers of pregnant and young forest musk deer were determined by performing the micro-agglutination test.The serum antibody titers of pregnant forest musk deer were more stable from 35th to 68th d after the third vaccination,and the serum antibody titers of four pregnant forest musk deer were maintained 25,25,25,and 24 on 68th d after the third vaccination.Young forest musk deer showed serum antibody titers which were obtained due to nursing.Young forest musk deer were administered the first intramuscular vaccine injection at an age of approximately 60 days due to a fall in maternal antibody titers.The serum antibody titers of young forest musk deer were higher after the third vaccination and maintained at approximately the same level until they were 137 days old.The maternal antibodies and the antibodies produced by young forest musk deer could be helpful for protecting the young musk deer from the infections of pathogenic Escherichia coli strains（serotypes O4,O26,and O139）for 137 days after birth（during the nursing period and the period when the forest musk deer were susceptible to diseases）.展开更多
基金research Grant from Beijing Science and Technology Program (No.00004105040311).
文摘A series of new O6-BG derivatives （14-21,23-30） were synthesized as inactivators of O6-Alkylguanine-DNA alkyltransferase （AGT）, and their ability to inhibit AGT was preliminary evaluated by MTT with O6-BG as the control. The result suggested compound 30 displayed a higher activity than O6-BG.
基金supported by the Science and Technology Planning Project of Fujian Province(No.2023Y4015)the Marine and Fishery Development Special Fund of Xiamen(No.23YYST064QCB36)the Natural Science Foundation of Fujian Province(No.2021J011210).
文摘The removal of ammonia nitrogen(NH_(4)^(+)-N)and bacteria from aquaculture wastewater holds paramount ecological and production significance.In this study,Pt/RuO_(2)/g-C_(3)N_(4)photocatalysts were prepared by depositing Pt and RuO_(2)particles onto g-C_(3)N_(4).The physicochemical properties of photocatalysts were explored by X-ray photoelectron spectroscopy(XPS),scanning electron microscopy(SEM),X-ray diffraction(XRD),and UV–vis diffuse reflectance spectrometer(UV–vis DRS).The photocatalysts were then applied to the removal of both NH_(4)^(+)-N and bacteria from simulated mariculture wastewater.The results clarified that the removals of both NH_(4)^(+)-N and bacteria were in the sequence of g-C_(3)N_(4)<RuO_(2)/g-C_(3)N_(4)<Pt/g-C_(3)N_(4)<Pt/RuO_(2)/g-C_(3)N_(4).This magnificent photocatalytic ability of Pt/RuO_(2)/g-C_(3)N_(4)can be interpreted by the transfer of holes from g-C_(3)N_(4)to RuO_(2)to facilitate the in situ generation of HClO from Cl^(−)in wastewater,while Pt extracts photogenerated electrons for H_(2)formation to enhance the reaction.The removal of NH_(4)^(+)-N and disinfection effect were more pronounced in simulated seawater than in purewater.The removal efficiency ofNH_(4)^(+)-N increases with an increase in pH of wastewater,while the bactericidal effect was more significant under a lower pH in a pH range of 6–9.In actual seawater aquaculture wastewater,Pt/RuO_(2)/g-C_(3)N_(4)still exhibits effective removal efficiency of NH_(4)^(+)-N and bactericidal performance under sunlight.This study provides an alternative avenue for removement of NH_(4)^(+)-N and bacteria from saline waters under sunlight.
文摘Solar-driven Fenton-like reactions are promising strategies for degrading pharmaceutical wastewater to address environmental challenges and antibiotic pollution.However,its efficacy is limited by suboptimal light absorption efficiency,rapid charge recombination,and inadequate interfacial charge transfer.In this study,an inorganic/organic S-scheme photo-Fenton system of pseudobrookite/carbon nitride(FTOCN)was synthesized via a hydrothermally coupled calcination process for the effective purification of tetracycline antibiotics under visible-light irradiation.The optimized FTOCN-2 heterostructure exhibits a significantly enhanced TC degradation capacity of 90%within 60 min.The rate constant of FTOCN-2 is 1.6 and 5.2 times greater than those of FTO and CN,respectively.Furthermore,FTOCN exhibits high antibacterial efficacy,highlighting its potential application in the purification of natural water.Measurements via a range of analytical techniques,including Kelvin probe force microscopy,density functional theory calculations,in situ X-ray photoelectron spectroscopy,and femtosecond transient absorption spectroscopy,corroborate the S-scheme mechanism.This study provides a novel perspective for the development of photo-Fenton systems with S-scheme heterojunctions for water purification.
基金supported by the National Key Research and Development Program of China(2021YFC2301700)the Natural Science Foundation of Heilongjiang Province,China(YQ2022C040)。
文摘Minks are highly susceptible to SARS-CoV-2,and have transmitted SARS-CoV-2 to humans.Oral immunization is one of the most promising strategies to prevent SARS-CoV-2 infection and transmission in minks.Here,we generated 3 recombinant rabies viruses(RABV),rERAG_(333E)/S6P,rERAG_(333E)/DS6P and rERAG_(333E)/BA2S6P,expressing the prefusion-stabilized SARS-CoV-2 spike protein of wild-type(S6P),δ(DS6P)or BA.2(BA2S6P)strain based on an oral rabies vaccine candidate(rERAG_(333E)).Oral or inactivated immunization of the 3 RABVs monovalent or trivalent were safe,and induced robust RABV neutralizing antibody and cross-antibody responses against the three SARS-CoV-2 in mice and minks.The challenge tests showed that 2 doses of rERAG_(333E)-S6P as an oral or inactivated vaccine completely protected mice against mouse-adapted SARS-CoV-2 infection in the upper and lower respiratory tracts,and largely prevented viral replication and lung damage caused by wild-type SARS-CoV-2infection in minks.Notably,we also confirmed that 2 doses of rERAG_(333E)-S6P as an oral or inactivated vaccine can largely protect minks against wild-type SARS-CoV-2 transmission via respiratory droplets.Our findings suggest that rERAG_(333E)-based COVID-19 vaccines appear to be suitable oral candidates to protect minks from SARS-CoV-2infection and transmission,and may serve as inactivated vaccines for further investigation in humans.
基金supported by the National Natural Science Foundation of China(Grant No.82200744)the Nanjing Science and Technology Development Funding(Grant No.YKK22159).
文摘Dent disease is a rare X-linked recessively inherited renal tubulopathy,caused by variants in the CLCN5(Dent disease type 1,DD1)and OCRL(Dent disease type 2,DD2)genes,and characterized by low molecular weight proteinuria,hypercalciuria,microscopic hematuria,or nephrocalcinosis.In the current study,we collected and analyzed clinical data and genetic testing results of 21 children diagnosed with Dent disease,who were hospitalized at the Department of Nephrology,Children's Hospital of Nanjing Medical University between January 2014 and August 2023,aiming to assist in the early diagnosis and treatment of these patients.Among the 21 patients,16(76.19%)had CLCN5 variants,and five(23.81%)had OCRL variants,and four of the variants were novel.All patients presented with low molecular weight proteinuria,14(66.67%)of whom had nephroticrange proteinuria.Eight patients underwent renal biopsies because of diagnostic uncertainty.We transfected the novel CLCN5 missense variant(p.G222R)and OCRL missense variant(p.I371T)plasmids into both HEK293 and HK-2 cells and found a significantly lower expression of the OCRL1 protein in the transfected cells than in the wild-type cells(P<0.05).Moreover,we observed an extremely skewed X-chromosome inactivation pattern in a female patient carrying the same novel CLCN5 variant,as assessed by the human androgen receptor gene assay.These findings provide insight into the clinical characteristics of Dent disease in Chinese patients and may shed light on its pathogenesis.
基金supported by the National Key Research and Development Program of China(2024YFC2309900 and 2021YFC2301200)the Zhejiang Plan for the Special Support for Top-notch Talents in China(2022R52029)+1 种基金the Fundamental Research Funds for the Central Universities(2022ZFJH003)the Hangzhou Global Scientific and Technological Innovation Center of Zhejiang University(KC2021ZY0B0002).
文摘Ionizing radiation presents an important solution for virus inactivation.However,its efficacy for severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)inactivation and the underlying mechanisms remain unclear.This study demonstrates radiosensitivity and radiation-induced biological changes in SARS-CoV-2 using 20 wild-type and mutant strains.The results show that 1.2 kGy of electron beam(E-beam)or 0.9 kGy of X-ray irradiation can eliminate 99.99%of SARS-CoV-2 particles.The Delta and various Omicron variants exhibit heightened sensitivity to radiation compared to the wild-type,showing nearly 99.99%inactivation efficiency at 1.0 and 0.8 kGy.The relationship between irradiation dose and the logarithmic reduction in virus load adheres to a dose-response model,characterized by extremely narrow windows.Spike(S)protein disruption,rather than the commonly accepted nucleic acid cleavage,is identified as the primary inactivation mechanism(triggering a conformation transition of S protein from pre-fusion to post-fusion with minimal impact on nucleic acid integrity).This study introduces the concept of targeting critical proteins in coronavirus inactivation,offering valuable insight for infectious coronavirus disease control and vaccine development.
基金National Institutes of Health grant F30DE030675(S.A.H.)National Institutes of Health grant T32DE007057(S.A.H.)+7 种基金University of Michigan Rackham Graduate School Pre-Candidate Research Grant(S.A.H.)University of Michigan Rackham Graduate School Candidate Research Grant(S.A.H.)National Institutes of Health grant R35DE034348(N.O.)National Institutes of Health grant R01DE026666(N.O.)National Institutes of Health grant R01DE030630(N.O.)National Institutes of Health grant R01DE029465(R.T.F.)Department of Defense grant W81XWH2010571(R.T.F.)National Institutes of Health grant P30 AR069620(The Michigan Integrative Musculoskeletal.Health Core Center).
文摘The cranial base synchondroses,comprised of opposite-facing bidirectional chondrocyte layers,drive anteroposterior cranial base growth.In humans,RUNX2 haploinsufficiency causes cleidocranial dysplasia associated with deficient midfacial growth.However,how RUNX2 regulates chondrocytes in the cranial base synchondroses remains unknown.To address this,we inactivated Runx2 in postnatal synchondrosis chondrocytes using a tamoxifen-inducible Fgfr3-creER(Fgfr3-Runx2cKO)mouse model.Fgfr3-Runx2cKO mice displayed skeletal dwarfism and reduced anteroposterior cranial base growth associated with premature synchondrosis ossification due to impaired chondrocyte proliferation,accelerated hypertrophy,apoptosis,and osteoclast-mediated cartilage resorption.Lineage tracing reveals that Runx2-deficient Fgfr3+cells failed to differentiate into osteoblasts.Notably,Runx2-deficient chondrocytes showed an elevated level of FGFR3 and its downstream signaling components,pERK1/2 and SOX9,suggesting that RUNX2 downregulates FGFR3 in the synchondrosis.This study unveils a new role of Runx2 in cranial base chondrocytes,identifying a possible RUNX2-FGFR3-MAPK-SOX9 signaling axis that may control cranial base growth.
基金supported by the Youth Innovation Promotion Association of CAS(2023350 to Xiaoxiao Gao).
文摘Dear Editor,Viruses of the genus Orthoebolavirus cause sporadic outbreaks of severe haemorrhagic fever,with case fatality rates ranging from 25%to 90%(Mahanty and Bray,2004).Six species of the virus(Orthoebolavirus zairense,sudanense,bundibugyoense,taiense,restonense,and bombaliense)have so far been identified(Biedenkopf et al.,2023).Among these,Orthoebolavirus zairense,commonly known as Ebola virus(EBOV),stands out as the most virulent.Given its high contagiousness and lethality,EBOV must be manipulated under biosafety level 4(BSL-4)conditions,as stipulated by the the People's Republic of China's list of human pathogenic microorganisms(National Health Commission of the People’s Republic of China,2023).Prior to being removed from a BSL-4 laboratory,it is imperative that infectious EBOV undergoes complete inactivation.Here we systematically evaluate viral thermostability under BSL-4 containment conditions,demonstrating EBOV's marked thermotolerance.
基金supported by the National Key Research and Development Program of China(2022YFE0139500)National Natural Science Foundation of China(32102105)+3 种基金Guangdong Basic and Applied Basic Research Foundation(2024A15150130262023A1515140136)Special Projects in Key Areas of Higher Education Institution in Guangdong Province(2024ZDZX2090)Guangdong Provincial Key Laboratory of Intelligent Food Manufacturing(2022B1212010015)。
文摘The natural curcumin-mediated photodynamic inactivation(PDI)was developed,and its inactivation potency against Fusarium graminearum in vitro and in vivo was systematically investigated by fluorescence probe assay,trypan blue staining,scanning electron microscope(SEM),confocal laser scanning microscopy(CLSM),etc.Results showed that under the irradiation of blue LED,the photosensitizer of curcumin was excited to generate massive reactive oxygen species(ROS)in the cells of F.graminearum,and the PDI completely inactivated their mycelia and spores under the treatment of 150μM curcumin and 10.8 J/cm^(2)irradiation.Further analysis found that the PDI ruptured the cellular microstructures,damaged the cell membrane by increasing its permeability and oxidizing the lipids,degraded the intracellular DNA and proteins inside the spores of F.graminearum.Meanwhile,the PDI also potently killed>99.99%spores of F.graminearum on maize under the treatment of 200μM curcumin and 10.8 J/cm^(2)irradiation.Moreover,the PDI suppressed the production of zearalenone(ZEN),and residual ZEN could not be detected after the storage of maize for 10 days.Therefore,this study systematically explored the inactivation efficiency of curcumin-mediated PDI against both the mycelia and spores of F.graminearum,which provides a valid and promising method to control the fungal hazards in grains.
基金supported by a grant from the National Key Research and Development Plans of China(No.2021YFD1800401)to Zhe Wangthe National Natural Science Foundation of China(No.32070128)to Zhe Wang+5 种基金the Shanghai Biomedical Science and Technology Support Special Project(No.21S11900200)to Zhe Wangthe National Natural Science Foundation of China(No.31472211)to ZhiBiao Yangthe Natural Science Foundation of Shanghai(No.21ZR1433900)to Zhibiao Yangthe National Key Research and Development Program of China(No.2022YFD1800805)to Yanjun Zhouthe Pinduoduo-China Agricultural University Research Fund(PC2024A01003)to Xiangfeng Wangthe Biomedicine and Technology Supporting Project of the Shanghai Science and Technology Innovation Plan(Grant No.22S11902200,China)to Tao Sun.
文摘Porcine epidemic diarrhea virus(PEDV)infection causes acute watery diarrhea in neonatal piglets,leading to substantial economic losses within the pig farming industry.This study demonstrates that clofazimine(CFZ)significantly inhibits PEDV replication in a dose-dependent manner in vitro,with negligible cytotoxicity.Findings from our time-of-addition assays indicate that CFZ effectively disrupts multiple stages of the viral infection cycle.Using a CoV-RdRp-Gluc reporter system,we evaluated the potency of CFZ against PEDV RNA-dependent RNA polymerase(RdRp),and determined a low IC50 value of 0.1364μM.Molecular docking studies further confirmed that CFZ has high binding affinity at the active sites of the spike protein and RdRp protein in PEDV.Transcriptome analysis of Vero E6 cells,with and without CFZ treatment,revealed a significant change in transcriptional activity at 8 h postinfection(hpi).Moreover,the simultaneous application of CFZ and nucleoside analogs showed enhanced the anti-PEDV effect of CFZ in vitro.Our study underscores the potential of CFZ as a viable therapeutic agent against PEDV.
基金Supported by the Shenzhen Science and Technology Program,No.JCYJ20220818102810022Shenzhen University Teaching Reform fund,No.JG2023166+1 种基金National Natural Science Foundation of China,No.12231018Science and Technology Innovation 2030-Major Project,No.2021ZD0201805.
文摘BACKGROUND Autoimmune encephalitis(AE)is a rare and recently described neuroinflammatory disease associated with specific autoantibodies.Anti-leucine-rich glioma inactivated 1(anti-LGI1)encephalitis is a rare but treatable type of AE discovered in recent years.Alzheimer’s disease(AD)is a degenerative brain disease and the most common cause of dementia.AD may undergo a series of pathological physiological changes in brain tissue 20 years before the onset of typical symptoms.The stage of mild cognitive impairment(MCI)that occurs during this process,known as MCI due to AD,is the earliest stage with clinical symptoms.MCI is typically categorized into two subtypes:Amnestic MCI(aMCI)and non-aMCI.CASE SUMMARY This report describes a patient with rapid cognitive impairment,diagnosed with anti-LGI1 antibody-mediated AE and aMCI,and treated at Peking University Shenzhen Hospital in March 2023.The patient was hospitalized with acute memory decline for more than 3 months.Both the cerebrospinal fluid and serum were positive for anti-LGI1 antibodies,biomarkers of AD coexisting in the patient’s cerebrospinal fluid.Following combination treatment with immunoglobulin therapy and glucocorticoid,plus inhibition of acetylcholinesterase,the patient’s cognitive function significantly improved.Throughout the 3-month follow-up period,a sustained improvement in cognitive function was observed.The results of serum anti-LGI1 antibody were negative.CONCLUSION This case has raised awareness of the possible interaction between AE and early AD(including MCI due to AD),and alerted clinicians to the possibility of concurrent rare and common diseases in patients presenting with cognitive impairment.
基金supported by the Swedish Research Council(2017-01286,2020-01840)the Swedish state under the agreement between the Swedish government and the county councils(ALF-agreement)(ALFGBG721581)+2 种基金the Gustaf V 80-years fund(FAI-2018-0466)the IngaBritt and Arne Lundberg Foundation(LU2017-0076)the Novo Nordisk Foundation(26844).
文摘Membrane-initiated estrogen receptorα(mERα)signaling has been shown to affect bone mass in murine models.However,it remains unknown which cell types mediate the mERα-dependent effects on bone.In this study,we generated a novel mouse model with a conditional C451A mutation in Esr1,which enables selective knockout of the palmitoylation site essential for the membrane localization of ERα(C451A^(f/f)).First,we used Runx2-Cre mice to generate Runx2-C451A^(f/f)mice with conditional inactivation of mERαsignaling in Runx2-expressing osteoblast lineage cells.No significant changes were observed in body weight,weights of estrogen-responsive organs,or serum concentrations of estradiol between female Runx2-C451A^(f/f)and homozygous C451A^(f/f)littermate controls.High-resolution microcomputed tomography analysis showed a consistent decrease in cortical bone mass in the tibia,femur,and vertebra L5 of Runx2-C451A^(f/f)mice and three-point bending analysis of humerus revealed an impaired mechanical bone strength in Runx2-C451A^(f/f)female mice compared to controls.Additionally,primary osteoblast cultures from mice lacking mERαsignaling showed impaired differentiation compared to controls.
基金supported by National Natural Science Foundation of China(Nos.22325602,22176060)Program of Shanghai Academic/Technology Research Leader(No.23XD1421000)+2 种基金Project supported by Shanghai Municipal Science and Technology Major Project(No.2018SHZDZX03)the Program of Introducing Talents of Discipline to Universities(No.B16017)Science and Technology Commission of Shanghai Municipality(No.20DZ2250400)。
文摘H_(2)O_(2)is an environmentally friendly oxidizing agent with minimal secondary pollution;however,its application has always been constrained by factors such as storage and transportation.In this study,we propose an innovative method for storing and releasing H_(2)O_(2)using hydrogels.Commercial hydrogels(sodium polyacrylate)can undergo swelling and absorb H_(2)O_(2)in aqueous solutions,and the swollen hydrogel can continuously release H_(2)O_(2)under osmotic pressure.And the characteristics of osmotic pressure drive ensure the recyclability of hydrogel for H_(2)O_(2)storage.Experimental results demonstrate that H_(2)O_(2)can stably exist within the hydrogel for an extended period,and this strategy helps to avoid explosion the risk and potential environmental hazards during the transportation of H_(2)O_(2).Finally,experiments confirm that the hydrogel controlled sustained release of H_(2)O_(2)is effective in both Fenton reactions and the process of bacterial inactivation.This work introduces new ideas for the storage of H_(2)O_(2),and the sustained release of H_(2)O_(2)may have significant implications in the fields of healthcare,environmental science,catalysis,and beyond.
基金supported by the Key Research and Development Program,Ministry of Science and Technology of the People’s Republic of China(Nos.2023YFC2606500,2023YFE0206500).
文摘Coronaviruses are single-stranded,positive-sense RNA enveloped viruses that have posed a significant threat to human health over the past few decades,particularly severe acute respiratory syndrome coronavirus(SARS-CoV),Middle East respiratory syndrome coronavirus(MERS-CoV),and SARS-CoV-2.These viruses have caused widespread infections and fatalities,with profound impacts on global economies,social life,and public health systems.Due to their broad host range in natural settings and the consequent high potential for zoonotic spillover events,a thorough investigation of the common viral mechanisms and the identification of druggable targets for pan-coronavirus antiviral development are of utmost importance.
基金Project(21271071)supported by the National Natural Science Foundation of ChinaProject(21306041)supported by the National Natural Science Young Foundation of China
文摘ZnO/Znml2O4 nanocomposites with heteronanostructures were successfully prepared by co-precipitation method. The as-prepared samples were characterized by HRTEM, TEM, XRD, BET, TG-DTA, and UV-Vis spectra techniques. The photoeatalytic activities of the as-prepared samples were evaluated by the photocatalytic degradation of methyl orange and inactivation of Escherichia coli in suspension under the irradiation of the simulated sunlight. The effects of compositions, calcination temperatures, concentration ofphotocatalysts and light source on the photocatalytic activities were systematically studied. The results show that when the concentration of ZnO/ZnA1204 photocatalyst with the starting Zn to Al molar ratio of 1：1.5 calcined at 600 ℃ is 1.0 g/L, the maximum photocatalytic degradation rate of 98.5% can be obtained in 50 min under the irradiation of the simulated sunlight. Under the same conditions, an inactivation rate of 99.8% for E.coli is achieved in 60 min.
文摘β-N-Acetyl-D-glucosaminidase （NAGase, EC.3.2.1.52） is chitinolytic enzymes and disintegrate dimmer and trimer a composition of oligomers of N-acetyl-β-D-glucosamine （NAG） into monomer. Prawn （P. vannamei） NAGase is involved in digestion and molting processes. Some pollutants in seawater affect the enzyme activity causing loss of the biological function of the enzyme, which affects the exuviating shell and threatens the survival of the animal. The effect of formaldehyde on prawn （P. vannamei） β-N-acetyl-D-glucosaminidase activity for the hydrolysis of pNP-NAG has been studied. The results show that formaldehyde, at appropriate concentrations, can lead to reversible inactivation of the enzyme, and the IC50 is estimated to be 1.05mol· L^-1. The inactivation mechanism obtained from Lineweaver-Burk plots shows that the inactivation of the enzyme by formaldehyde belongs to the competitive type. The inactivation kinetics of the enzyme by formaldehyde has been studied using the progress-of-substrate-reaction method described by Tsou, and the rate constants have been determined. The results show that k＋0 is much larger than k-0, indicating the free enzyme molecule is fragile in the formaldehyde solution.
基金the Natural Scientific Foundation of China (NSFC3962526)National High-Technology Project-863 (102-10-01-04)
文摘AIM: To develop and optimize cDNA representational difference analysis (cDNA RDA) method and to identify and clone garlic up-regulated genes in human gastric cancer (HGC) cells. METHODS: We performed cDNA RDA method by using abundant double-stranded cDNA messages provided by two self-constructed cDNA libraries (Allitridi-treated and paternal HGC cell line BGC823 cells cDNA libraries respectively). Bam H I and Xho I restriction sites harbored in the library vector were used to select representations. Northern and Slot blots analyses were employed to identify the obtained difference products. RESULTS: Fragments released from the cDNA library vector after restriction endonuclease digestion acted as good marker indicating the appropriate digestion degree for library DNA. Two novel expressed sequence tags (ESTs) and a recombinant gene were obtained. Slot blots result showed a 8-fold increase of glia-derived nexin/protease nexin 1 (GDN/PN1) gene expression level and 4-fold increase of hepatitis B virus x-interacting protein (XIP) mRNA level in BGC823 cells after Allitridi treatment for 72h. CONCLUSION: Elevated levels of GDN/PN1 and XIP mRNAs induced by Allitridi provide valuable molecular evidence for elucidating the garlic's efficacies against neurodegenerative and inflammatory diseases. Isolation of a recombinant gene and two novel ESTs further show cDNA RDA based on cDNA libraries to be a powerful method with high specificity and reproducibility in cloning differentially expressed genes.
文摘By 30% - 60% (NH4)(2)SO4 fractional precipitation, anion-exchange chromatography on DEAE-Sepharose CL-6B, gel filtration on Sephacryl S-200 and anion-exchange chromatography on Waters AP-1 column (Protein(PM)-Pak DEAE 15HR), a proteinase which can inactivate soybean trypsin inhibitor (STI) was purified from mung bean ( Vigna rabiata (L.) Wilezek) sprouts. Its molecular weight was estimated to be 29.8 kD by SDS-PAGE, and its K-m and V-max for STI were 769.2 N-alpha -benzoyl-L-arginine ethyl ester BAEE/mL and 115.3 BAEE . mL(-1) . min(-1) respectively. This proteinase was stable at temperatures lower than 50 degreesC and pH 6.5 - 8.5, and 90.91% STI activity of defatted soybean powder was inactivated by this preparation, with proteolytic activity 5 000 BAEE/mL at 50 degreesC and pH 8.0 in 4 h.
基金Supported by the Government Funded Clinical Medicine Eexcellent Talent Training and Basic Research Project PlanNatural Science Foundation of Hebei(No.H2019423092)+2 种基金Higher Education Science and Technology Research Project of Hebei(No.ZD2016056)Postgraduate Innovation Ability Development Project of Hebei Education Department(No.CXZZBS2019159)Basic Research Business Expenses of Provincial Universities of Hebei University of Chinese Medicine Project of Excellent Student Research Capacity Improvement(No.YXZ2019001)。
文摘OBJECTIVE: To investigate the effect of Danggui Buxue Tang(DBT), a decoction from Traditional Chinese Medicine, on bleomycin-induced pulmonary fibrosis in rats, and to propose the possible underlying mechanism.METHODS: Forty male Sprague-Dawley rats were randomly divided into sham group, model group,prednisone group and DBT group. Pulmonary fibrosis rat model was established by intratracheal injection with bleomycin. Body weight and lung index were monitored. Histopathologic examination and collagen deposition were determined using Hematoxylin and eosin(HE) and Masson's trichrome staining. Immunohistochemistry staining was applied to observe the expression of alpha-smooth muscle actin(α-SMA). m RNA expression of α-SMA,collagen Ⅰ and collagen Ⅲ were measured by realtime fluorescence quantitative PCR(RT-q PCR). Inflammatory cytokines, including tumor necrosis factor alpha(TNF-α), interleukin-6(IL-6) and IL-1β in serum were detected by Enzyme-linked immunosorbent assay. Alkali hydrolysis method was conducted to investigate the content of hydroxyproline(HYP). Transforming growth factor-β1(TGF-β1),Smad3 and plasminogen activator inhibitor-1(PAI-1) protein level were examined by Western blot assay.RESULTS: DBT significantly reduced the severity of bleomycin-induced pulmonary fibrosis and inflammation as indicated by minimizing the lost of weight, and by lowering the levels of lung index, inflammation score, Ashcroft score, collagen volume fraction(%), HYP, α-SMA, collagen Ⅰ, collagen Ⅲ,TNF-α, IL-6, IL-1β, TGF-β1, Smad3 and PAI-1, consistent with the effect of prednisone.CONCLUSION: Our findings suggest that DBT is able to ameliorate the pulmonary fibrosis, the possible mechanism may involve inhibition of pulmonary inflammation and collagen deposition, possibly via suppressing TGF-β1/Smad3/PAI-1 signaling pathway.
基金Supported by Youth Foundation of Education Department in Sichuan Province(07ZB060)Scientific and Technological Supporting Project in Science and Technology Bureau of Sichuan Province(2009SZ0228)~~
文摘A multivalent inactivated Escherichia coli vaccine for forest musk deer by using serotypes O4,O26,and O139 with Al（OH）3 adjuvant was prepared.The vaccine did not cause any adverse reactions in forest musk deer.The immunogenic effects of the vaccine were experimentally investigated in pregnant and young forest musk deer.The serum antibody titers of pregnant and young forest musk deer were determined by performing the micro-agglutination test.The serum antibody titers of pregnant forest musk deer were more stable from 35th to 68th d after the third vaccination,and the serum antibody titers of four pregnant forest musk deer were maintained 25,25,25,and 24 on 68th d after the third vaccination.Young forest musk deer showed serum antibody titers which were obtained due to nursing.Young forest musk deer were administered the first intramuscular vaccine injection at an age of approximately 60 days due to a fall in maternal antibody titers.The serum antibody titers of young forest musk deer were higher after the third vaccination and maintained at approximately the same level until they were 137 days old.The maternal antibodies and the antibodies produced by young forest musk deer could be helpful for protecting the young musk deer from the infections of pathogenic Escherichia coli strains（serotypes O4,O26,and O139）for 137 days after birth（during the nursing period and the period when the forest musk deer were susceptible to diseases）.
