Background Plant tissue culture has emerged as a tool for improving cotton propagation and genetics,but recalcitrance nature of cotton makes it difficult to develop in vitro regeneration.Cotton’s recalcitrance is inf...Background Plant tissue culture has emerged as a tool for improving cotton propagation and genetics,but recalcitrance nature of cotton makes it difficult to develop in vitro regeneration.Cotton’s recalcitrance is influenced by genotype,explant type,and environmental conditions.To overcome these issues,this study uses different machine learning-based predictive models by employing multiple input factors.Cotyledonary node explants of two commercial cotton cultivars(STN-468 and GSN-12)were isolated from 7–8 days old seedlings,preconditioned with 5,10,and 20 mg·L^(-1) kinetin(KIN)for 10 days.Thereafter,explants were postconditioned on full Murashige and Skoog(MS),1/2MS,1/4MS,and full MS+0.05 mg·L^(-1) KIN,cultured in growth room enlightened with red and blue light-emitting diodes(LED)combination.Statistical analysis(analysis of variance,regression analysis)was employed to assess the impact of different treatments on shoot regeneration,with artificial intelligence(AI)models used for confirming the findings.Results GSN-12 exhibited superior shoot regeneration potential compared with STN-468,with an average of 4.99 shoots per explant versus 3.97.Optimal results were achieved with 5 mg·L^(-1) KIN preconditioning,1/4MS postconditioning,and 80%red LED,with maximum of 7.75 shoot count for GSN-12 under these conditions;while STN-468 reached 6.00 shoots under the conditions of 10 mg·L^(-1) KIN preconditioning,MS with 0.05 mg·L^(-1) KIN(postconditioning)and 75.0%red LED.Rooting was successfully achieved with naphthalene acetic acid and activated charcoal.Additionally,three different powerful AI-based models,namely,extreme gradient boost(XGBoost),random forest(RF),and the artificial neural network-based multilayer perceptron(MLP)regression models validated the findings.Conclusion GSN-12 outperformed STN-468 with optimal results from 5 mg·L^(-1) KIN+1/4MS+80%red LED.Application of machine learning-based prediction models to optimize cotton tissue culture protocols for shoot regeneration is helpful to improve cotton regeneration efficiency.展开更多
In this study,Mg-based composites,by the addition of ZnO,Ca_(2)ZnSi_(2)O_(7),Ca_(2)MgSi_(2)O_(7),and CaSiO_(3)as bioactive agents,were fabricated using friction stir processing.The microstructure and in vitro assessme...In this study,Mg-based composites,by the addition of ZnO,Ca_(2)ZnSi_(2)O_(7),Ca_(2)MgSi_(2)O_(7),and CaSiO_(3)as bioactive agents,were fabricated using friction stir processing.The microstructure and in vitro assessment of bioactivity,biodegradation rate,and corrosion behavior of the resultant composites were investigated in simulated body fluid(SBF).The results showed that during the immersion of composites in SBF for 28 d,due to the release of Ca^(2+)and PO_(4)^(3-)ions,hydroxyapatite(HA)crystals with cauliflower shaped morphology were deposited on the surface of composites,confirming good bioactivity of composites.In addition,due to the uniform distribution of bioceramic powders throughout Mg matrix,grain refinement of the Mg matrix,and uniform redistribution of secondary phase particles,the polarization resistance increased,and the biodegradation rate of composites significantly reduced compared to monolithic Mg matrix.The polarization corrosion resistance of Mg-ZnO increased from 0.216 to 2.499 kΩ/cm^(2)compared to monolithic Mg alloy.Additionally,Mg-ZnO composite with the weight loss of 0.0217 g after 28 d immersion showed lower weight loss compared to other samples with increasing immersion time.Moreover,Mg-ZnO composite with the biodegradation rate of 37.71 mm/a exhibited lower biodegradation rate compared to other samples with increasing immersion time.展开更多
Studying the skin care efficacy of recombinant humanized collagen based on in vitro level.The stability of the recombinant humanized collagen was first analyzed by treating at different temperatures,then its skincare ...Studying the skin care efficacy of recombinant humanized collagen based on in vitro level.The stability of the recombinant humanized collagen was first analyzed by treating at different temperatures,then its skincare efficacy based on in vitro level was evaluated by detecting the inhibition rate of elastase,the inhibition rate of collagenase,the protein content of type I collagen in human fibroblasts,the inhibition of reactive oxygen species(ROS)with human keratinocytes,and the effects of the recombinant humanized collagen on the expression of hyaluronic acid(HA),filaggrin(FLG)and transglutaminase 1(TGM1)in keratinocytes.The results showed that recombinant humanized collagen was able to maintain stability at temperatures below 70℃.With regard to its skincare efficacy,recombinant humanized collagen could inhibit elastase and collagenase activities and promote the increase of type I collagen content in human fibroblasts.It also showed good inhibition of ROS in keratinocytes in vitro and could increase the expression of HA,FLG,and TGM1 in keratinocytes.In short,the recombinant humanized collagen exhibited a favourable skin care effect in vitro level.This study proved that it has potential firming,anti-wrinkle,moisturizing,and repairing efficacy,and is a valuable cosmetic raw material.展开更多
As an important wild blueberry resource,Vaccinium uliginosum has attracted more and more attention.At present,the wild resources are under destruction.The conservation of wild Vaccinium uliginosum resources is imminen...As an important wild blueberry resource,Vaccinium uliginosum has attracted more and more attention.At present,the wild resources are under destruction.The conservation of wild Vaccinium uliginosum resources is imminent.However,there are few researches on the protection and preservation of its germplasm resources.In vitro preservation is an important method for germplasm conservation.In this study,one strain of wild Vaccinium uliginosum was used as material.The effects of temperature(25℃,15℃,10℃,or 0℃),media(WPM,1/2WPM or 1/3WPM),medium supplements(sorbitol or mannose),and photoperiod(8,10,12,or 14 h•d^(-1))on the growth,survival rate and rejuvenation rate of the plantlets were studied.The physiological changes of plantlets during preservation were analyzed.Methylation-sensitive amplified polymorphism(MSAP)analysis of genomic DNA methylation of plantlets was carried out to explore the genetic stability of the plantlets after preservation.The research results provided a theoretical basis for the germplasm preservation of Vaccinium uliginosum.展开更多
The in vitro maturation, fertilization and development of bovine ovary oocytes in two differentcultural systems A and B were studied under the conditions of 38.5℃,5%CO2,95%air and 100%humidity.The maturation rates we...The in vitro maturation, fertilization and development of bovine ovary oocytes in two differentcultural systems A and B were studied under the conditions of 38.5℃,5%CO2,95%air and 100%humidity.The maturation rates were 94.5%and 91.3%,respectively,and the difference wasextremely significant.Frozen semen were thawed and sperm were capacitated with three kinds ofcapacitation agents for fertilization.The pronucleus rates were 76%,65%-68%and 62%respectively.The rates of embryos developed to morula and blastocyst were 19%,16% and 17%respectively.The developmental rates of embryos cocultured with bovine oviductal epithelium cellsand bovine granulosa cells were 25% and 23.4% respectively,with no significant difference. Freshembryos were transplanted into 15 recipicns,and three of them were pregnant and calves were bornin 1990 and 1991.The pregnant rate was 20%.The emryos developed faster before 8-cell stage andslower after 8-cell stage,in vitro than in vivo.展开更多
The antioxidant effects of quercetin were studied in vitro and in vivo.In vitro,vitamin C was used as a positive control to evaluate the antioxidant capacity of quercetin in three aspects:scavenging free radicals,prot...The antioxidant effects of quercetin were studied in vitro and in vivo.In vitro,vitamin C was used as a positive control to evaluate the antioxidant capacity of quercetin in three aspects:scavenging free radicals,protecting biological macromolecules and the total reducing power.In vivo,a total of 240 AA broilers(1-day age)with similar body weight were randomly divided into four groups with six replicates in each group,and 10 broilers in each replicate.The four groups were fed with corn-soybean basal diet supplemented with 0.00%,0.02%,0.04%and 0.06%quercetin to study its effects on antioxidant indexes of AA broilers,and to explore the optimal dose of quercetin as a dietary additive.The results showed that quercetin scavenged superoxide anion,hydroxyl radical and 1,1-diphenyl-2-picrylhydrazyl(DPPH)in vitro,the scavenging effects of quercetin on O_(2)-and•OH first increased and then decreased with the increase of the concentrations(P<0.01),and its maximum scavenging effect was observed at concentrations of 40 and 300 mg·L^(-1).The scavenging effects of quercetin on DPPH was increased constantly with increasing concentrations.The scavenging effect of quercetin on three free radicals was DPPH>•OH>O_(2)-.The inhibition of vitelline lipoprotein peroxidation by quercetin was increased with increasing concentrations(P<0.01)and the inhibitory effect was higher than that of vitamin C.The inhibition of red blood cell hemolysis by quercetin was increased with increasing concentrations at 0.05-1.25 mg·L^(-1)(P<0.01);however,the inhibition tended to decrease when the concentration was too high(31.25 mg·L^(-1)),and the inhibitory effect was higher than that of vitamin C.The inhibition of mitochondrial expansion by quercetin was increased with increasing concentrations,according to the degree of mitochondrial expansion at 60 min,the integrity of mitochondria in the experimental groups was significantly higher than that in the model group(P<0.01).The total reducing power of quercetin was increased with increasing concentrations(P<0.01);however,the total reducing power was less than that of vitamin C.In vivo,malondialdehyde(MDA)and nitric oxide(NO)were significantly decreased with increasing quercetin(P<0.01).Quercetin supplementation had no effect on the content of lipid peroxidation(LPO)in livers(P>0.05);however,superoxide dismutase(SOD)activity was significantly increased,whereas glutathione peroxidase(GSH-Px)and catalase(CAT)activities were significantly decreased in livers with increasing quercetin(P<0.05).These results suggested that quercetin exhibited strong antioxidant effects in vitro and in vivo.展开更多
Three methods were adopted in culture spermatogoniums of newly born calf in vitro,such as enzymatic digestion and percoll density gradient centrifugation（MethodⅠ）,tubular fragments culture（MethodⅡ）and tissue cul...Three methods were adopted in culture spermatogoniums of newly born calf in vitro,such as enzymatic digestion and percoll density gradient centrifugation（MethodⅠ）,tubular fragments culture（MethodⅡ）and tissue culture（MethodⅢ）,and cultural behaviors of cells were observed.The results showed that typical spermatogonium colonies appeard at 144 h of culture by enzymatic digestion-percoll density gradient centrifugation method and tubular fragments culture method,2.5%FBS kept the characteristics of spermatogonium stem cell better than others,produced more mass clones,and FBS of more than 2.5%concentration benefited spermatogonium differentiation and the number of colonies was significantly affected by FBS concentration.After 1 week of culture in method Ⅲ,the diameter of lumens and quantity of sertoli’s cells in tubal wall increased obviously,lumen of seminiferous tubules appeared.Sertoli’s cells kept constant and the number of spermatogoniums decreased obviously after 2 weeks of culture.展开更多
Mg-6%Zn-10%β-Ca3(PO4)2 composite was prepared through powder metallurgy methods with different chitosan coatings on its surface. The properties of the chitosan coatings on the surface of Mg-6%Zn-10%β-Ca3(PO4)2 compo...Mg-6%Zn-10%β-Ca3(PO4)2 composite was prepared through powder metallurgy methods with different chitosan coatings on its surface. The properties of the chitosan coatings on the surface of Mg-6%Zn-10%β-Ca3(PO4)2 composite, such as the adhesion ability, the corrosion behavior and the cytotoxicity properties, were investigated, and the microstructure of the chitosan coating was observed by scanning electron microscope(SEM). The results show that chitosan coating improves the corrosion resistance of the magnesium composite specimens significantly. Mg-6%Zn-10%β-Ca3(PO4)2 composite specimens exhibit good corrosion resistance and low p H values in simulated body fluid(SBF) at 37 °C in the immersion test with 7-layer chitosan coating whose relative molecular mass is 30×104 Da. The cytotoxicity tests indicate that Mg-6%Zn-10%β-Ca3(PO4)2 with chitosan coating is nontoxic with a cytotoxicity grade of zero against L-929 cells, which is better than that of uncoated composites.展开更多
OBJECTIVE To explore mecha⁃nisms of imperatorin on regulating P-glycoprotein(P-gp)in blood-brain barrier(BBB)based on net⁃work pharmacology combined with in vitro experi⁃ment.METHODS Drug targets were predicted using ...OBJECTIVE To explore mecha⁃nisms of imperatorin on regulating P-glycoprotein(P-gp)in blood-brain barrier(BBB)based on net⁃work pharmacology combined with in vitro experi⁃ment.METHODS Drug targets were predicted using the Pharmapper and Swiss targets data⁃bases;disease targets were obtained through the Genecards database;intersections between drugs and disease targets were screened by Cytoscape software;the obtained core targets were used to construct protein-protein interaction(PPI)network,gene ontology(GO)functions,and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.The effects of imperatorin(20,50,100μmol·L^(-1))on P-gp activity were monitored in hCMEC/D3 in vitro BBB model,and the effects of imperatorin on the expression of target proteins were verified using Western blot method.RESULTS 55 drug targets and 3102 disease targets were obtained from the network pharmacology screening,and 37 core targets were obtained after the combination.Enrichment analysis showed that core targets were closely related to chemical synaptic trans⁃mission regulation,neurotransmitter receptor activity,protein kinase regulation activity,G proteincoupled receptor signaling pathway,neural active ligand receptor interaction pathway,PI3K-Akt sig⁃naling pathway,VEGF signaling pathway,etc..In vitro experimental validation suggested that all tested concentration groups of imperatorin signifi⁃cantly reduced the activity and expression of P-gp,which were achieved by significantly downregu⁃lating the phosphorylation levels of PI3K and Akt,and repressing the expression of VEGFR2 pro⁃tein.CONCLUSION Network pharmacology was used to predict the core targets and signaling pathways of imperatorin on regulating P-gp in BBB and relevant validation was conducted through in vitro experiments,providing a refer⁃ence basis for further exploration of the mecha⁃nisms of imperatorin on regulating P-gp in BBB.展开更多
Influenza virus infection is a global public health issue.The effectiveness of antiviral agents for influenza has been limited by the emergence of drugresistant virus strains.Therefore,there is an urgent need to ident...Influenza virus infection is a global public health issue.The effectiveness of antiviral agents for influenza has been limited by the emergence of drugresistant virus strains.Therefore,there is an urgent need to identify novel antiviral therapies.Our previous studies have found that Cryptoporus volvatus extract could potently inhibit influenza virus replication in vitro and in vivo.However,the effective component of Cryptoporus volvatus which mediated the antiviral activity hasn′t been identified.Here,we identified a novel anti-influenza molecule,cryptoporic acid E(CAE),from Cryptoporus volvatus.Our results showed that CAE had broad-spectrum anti-influenza activity against 2009 pandemic strain A/Beijing/07/2009(H1N1/09),seasonal strain A/Jiangxi/262/05(H3N2),mouse adapted strains A/WSN/33(H1N1)and A/PR8/34(H1N1).We further investigated the mode of CAE action,and found that CAE directlyattenuated influenza virus infectivity.Time-course-analysis indicated that CAE exerted its inhibition mainly at middle stage of the replication cycle of influenza virus.Subsequently,we confirmed that CAE blocked virus RNA replication and transcription in MDCK cells and CAE repressed influenza virus RNA polymerase activity.In addition,we found that CAE impaired influenza virus infectivity by directly targeting virus particles.Our data suggest that CAE is a major effective component of Cryptoporus volvatus and might be a potential candidate for the development of a new anti-influenza virus therapy.展开更多
In this paper, several factors that affect the efficiency of in vitro adventitious bud regeneration and Agrobacterium tumefaciens-mediated transformation of F. vesca were studied. The results showed that F. vesca see...In this paper, several factors that affect the efficiency of in vitro adventitious bud regeneration and Agrobacterium tumefaciens-mediated transformation of F. vesca were studied. The results showed that F. vesca seeds germination rate was the highest while seeds were cultured in water, and the germination rate was the lowest while seeds were cultured on MS medium supplemented with hormone; the germination rates that seeds cultured on two and three layers filter paper were higher than that seeds cultured on four and five layers filter paper. In vitro adventitious regeneration efficiency was affected by different explants types. The significant difference was existed between petioles and leaves. When using the same type explants, in vitro adventitious buds regeneration rate and the average number of buds per explant between Ruegen （RE） and Yellow Wonder （YW） had no significant difference. RE to Agrobacterium tumefaciens was more sensitive than YW. Using seedling leaves of RE and YW as materials, an efficient Agrobacterium-mediated transformation system was developed. In this system, the concentration of bacteria was OD600=0.5, the explants were immersed in bacteria broth for 9 min, the co-cultured time was 2 days, and had no pre-cultured time. The percentage of explants with resistant buds of RE and YW was compared. The putative transformed plants were confirmed by PCR.展开更多
To investigate the effects and mechanisms of the benzimidazole carbamate and benmidine drugs on Cysticerci cellulosae and choose effective drugs on Cysticerci cellulosae, the membrane metabolism of Cysticerci cellulos...To investigate the effects and mechanisms of the benzimidazole carbamate and benmidine drugs on Cysticerci cellulosae and choose effective drugs on Cysticerci cellulosae, the membrane metabolism of Cysticerci cellulosae in vitro was tested after three kinds of drugs which were used respectively. The indexes included the contents of lipids, the contents of SA and the changes of the membrane fluidity. The results showed that oxfendazole could inhibit the membrane metabolism of immature and mature Cysticerci cellulosae in vitro, and albendazole only inhibited the mature one, while thibendimidine neither acted on the immature nor mature one.展开更多
Collected from the broiler cecal contents, bacteria were isolated and purified. These isolated and purified Lactobacilli were detected by acidity and bile salt tolerant test, then three better tolerant strains were sc...Collected from the broiler cecal contents, bacteria were isolated and purified. These isolated and purified Lactobacilli were detected by acidity and bile salt tolerant test, then three better tolerant strains were screened out. These three strains were conducted to observe the morphous through microscope, perform Gram stain, and to identify their physiological and biochemical characteristics. The results showed that L1 was L.casei, L2 was L.gasseri and L3 was L.graminis. With Oxford Cup method, these three Lactobacilli were detected for their bacteriostasis. Results showed that L1, L2 and L3 strains had strong antibacterial activities to E. coli and Salmonella.展开更多
The histological observation was experimentally conducted on in vitro cultured mouse embryonic myocardium cells and myocardiumoid cell mass. The mouse embryo tissue were cultured and regular pulsatile myocardiumoid ti...The histological observation was experimentally conducted on in vitro cultured mouse embryonic myocardium cells and myocardiumoid cell mass. The mouse embryo tissue were cultured and regular pulsatile myocardiumoid tissue could be found. During in vitro culture, the myofilament bundles in the cell were gradually increasing and strongly connectted each other with embryonic age and there were loose muscle fibers initially and intercalated discs were close to each other. The lose myofilament bundles were developed in muscle fibers with age and the distance between intercalated discs was enlarged. There were myofilamentoid structure in inactive cells and filament peripherily.展开更多
Cyclamen leaves and petioles explants were cultured on MS media supplemented with different concentrations of 2, 4-dichlorophenoxyacetic acid (2, 4-D) or 1-naphthaleneacetic acid (NAA) to induce callus. The effect of ...Cyclamen leaves and petioles explants were cultured on MS media supplemented with different concentrations of 2, 4-dichlorophenoxyacetic acid (2, 4-D) or 1-naphthaleneacetic acid (NAA) to induce callus. The effect of 2, 4-D on shoot regeneration was also studied. Either in media containing 2, 4-D or in media containing NAA, callus was observed, but the quality or quantity of callus induced by 2, 4-D or NAA were different. The callus induced by 2, 4-D was white, compact and having powerful multiplication capacity. The callus was inclined to browning then was poorly organogenetic. While the callus induced by NAA was yellowish in appearance. It was pultaceous and proliferated bradytelicly. The callus usually can give rise to many shoots. But the frequency of inducing callus of 2, 4-D is higher than that of NAA. The regenerative plantlets derived from the callus respectively induced by 2, 4-D or NAA were transferred into rooting medium. The frequency of rooting were no difference.展开更多
Batch cultures of mixed rumen micro-organisms were conducted to evaluate the effects of encapsulated yeast(+EY)and encapsulated enzyme(+EE)using plant proteins(barley and oats grain)on rumen fermentation in vitro,inve...Batch cultures of mixed rumen micro-organisms were conducted to evaluate the effects of encapsulated yeast(+EY)and encapsulated enzyme(+EE)using plant proteins(barley and oats grain)on rumen fermentation in vitro,investigate the abilities of encapsulated yeast and encapsulated enzyme to prevent rumen digestion in vitro.Treatments of the study were the control,+EY,+EE products(3.33 mg·mL^(-1) of the incubation medium),unencapsulated yeast(-EY)and enzyme(-EE)products(0.17 and 0.17μL·mL^(-1) of the incubation medium,respectively).+EY group increased dry matter disappearance(DMD,P<0.01)and the total volatile fatty acids(TVFA,P<0.01)at 3 h of the incubation compared with the control,regardless of encapsulation of yeast.Gas production(GP)of+EY group was higher(P=0.05,29.94 mL·mL^(-1) organic matter,OM)than that of the control(25.08 mL·g^(-1) OM)at 3 h of the incubation.Supplementation+EY increased DMD(P=0.04,0.394 vs 0.352,respectively)and acetic proportion(P=0.04,52.6 vs 49.8 mol•100 mL^(-1),respectively)at 6 h of the incubation and increased A:P ratio(P<0.01,3.11 and 2.86,respectively)at 24 h of the incubation,as compared to unencapsulation of yeast.Supplementation of enzyme had higher(P≤0.04)GP,DMD and TVFA at 3 and 6 h of the incubation compared with the control,regardless of encapsulation.Moreover,the addition of+EE produced greater GP at 6(P<0.01,92.35 vs 78.21 mL·g^(-1) OM,respectively),12(218.47 vs 159.18 mL·g^(-1) OM)and 24 h(380.97 vs 297.78 mL·g^(-1) OM,respectively)of the incubation,higher DMD(0.347 vs 0.313,respectively)at 3 h of the incubation as compared to-EE group.The study showed that the encapsulation might protect part of yeast and enzyme from releasing to the rumen throughout the digestion in vitro,resulting in higher or no difference of rumen fermentation parameters compared with unencapsulated groups at any incubation times.In comparison with-EY and-EE,the higher rumen fermentation parameters at the early incubation time were observed,which could be attributed to the higher concentration of yeast or enzyme.However,regardless of the encapsulation,the results indicated that both yeast and enzyme only improved the speed rather than the extent of rumen fermentation in vitro.展开更多
Introduction Cancer is an attractive target of gene therapy and currently represents the disease in most clinical trials[1]. Strategies for cancer gene therapy include: (1) stimulation of immune responses to tumor cel...Introduction Cancer is an attractive target of gene therapy and currently represents the disease in most clinical trials[1]. Strategies for cancer gene therapy include: (1) stimulation of immune responses to tumor cells,(2) delivery of specific enzymes展开更多
The present investigation was undertaken to find out the best medium composition for medium-term preservation of disease free potato shoot tips in in vitro system.Thirteen potato genotypes and fourteen treatments were...The present investigation was undertaken to find out the best medium composition for medium-term preservation of disease free potato shoot tips in in vitro system.Thirteen potato genotypes and fourteen treatments were taken under consideration for the present experiment.Among fourteen treatments,mannitol and sorbitol containing media proved to be the best for medium-term preservation of potato shoot tips.High concentration of mannitol delayed root formation.展开更多
Biomaterials for restoration or replacement of diseased tissues may have any origin.The major characteristic for biomaterials is biocompatibility.All biomaterials,used in medicine (dentistry,in particular),interreact ...Biomaterials for restoration or replacement of diseased tissues may have any origin.The major characteristic for biomaterials is biocompatibility.All biomaterials,used in medicine (dentistry,in particular),interreact with the organism tissues.And the changes occur both in the materials and the organism tissues.It is considered that there are no "inert biomaterials." The number of allergic diseases and complications is constantly growing all over the world,taking an important place in the structure of infectious and noninfectious pathology[1].Pollen,household,epidermal,and food-borne allergens,and haptens are the most frequent sources of sensibilization.展开更多
文摘Background Plant tissue culture has emerged as a tool for improving cotton propagation and genetics,but recalcitrance nature of cotton makes it difficult to develop in vitro regeneration.Cotton’s recalcitrance is influenced by genotype,explant type,and environmental conditions.To overcome these issues,this study uses different machine learning-based predictive models by employing multiple input factors.Cotyledonary node explants of two commercial cotton cultivars(STN-468 and GSN-12)were isolated from 7–8 days old seedlings,preconditioned with 5,10,and 20 mg·L^(-1) kinetin(KIN)for 10 days.Thereafter,explants were postconditioned on full Murashige and Skoog(MS),1/2MS,1/4MS,and full MS+0.05 mg·L^(-1) KIN,cultured in growth room enlightened with red and blue light-emitting diodes(LED)combination.Statistical analysis(analysis of variance,regression analysis)was employed to assess the impact of different treatments on shoot regeneration,with artificial intelligence(AI)models used for confirming the findings.Results GSN-12 exhibited superior shoot regeneration potential compared with STN-468,with an average of 4.99 shoots per explant versus 3.97.Optimal results were achieved with 5 mg·L^(-1) KIN preconditioning,1/4MS postconditioning,and 80%red LED,with maximum of 7.75 shoot count for GSN-12 under these conditions;while STN-468 reached 6.00 shoots under the conditions of 10 mg·L^(-1) KIN preconditioning,MS with 0.05 mg·L^(-1) KIN(postconditioning)and 75.0%red LED.Rooting was successfully achieved with naphthalene acetic acid and activated charcoal.Additionally,three different powerful AI-based models,namely,extreme gradient boost(XGBoost),random forest(RF),and the artificial neural network-based multilayer perceptron(MLP)regression models validated the findings.Conclusion GSN-12 outperformed STN-468 with optimal results from 5 mg·L^(-1) KIN+1/4MS+80%red LED.Application of machine learning-based prediction models to optimize cotton tissue culture protocols for shoot regeneration is helpful to improve cotton regeneration efficiency.
文摘In this study,Mg-based composites,by the addition of ZnO,Ca_(2)ZnSi_(2)O_(7),Ca_(2)MgSi_(2)O_(7),and CaSiO_(3)as bioactive agents,were fabricated using friction stir processing.The microstructure and in vitro assessment of bioactivity,biodegradation rate,and corrosion behavior of the resultant composites were investigated in simulated body fluid(SBF).The results showed that during the immersion of composites in SBF for 28 d,due to the release of Ca^(2+)and PO_(4)^(3-)ions,hydroxyapatite(HA)crystals with cauliflower shaped morphology were deposited on the surface of composites,confirming good bioactivity of composites.In addition,due to the uniform distribution of bioceramic powders throughout Mg matrix,grain refinement of the Mg matrix,and uniform redistribution of secondary phase particles,the polarization resistance increased,and the biodegradation rate of composites significantly reduced compared to monolithic Mg matrix.The polarization corrosion resistance of Mg-ZnO increased from 0.216 to 2.499 kΩ/cm^(2)compared to monolithic Mg alloy.Additionally,Mg-ZnO composite with the weight loss of 0.0217 g after 28 d immersion showed lower weight loss compared to other samples with increasing immersion time.Moreover,Mg-ZnO composite with the biodegradation rate of 37.71 mm/a exhibited lower biodegradation rate compared to other samples with increasing immersion time.
文摘Studying the skin care efficacy of recombinant humanized collagen based on in vitro level.The stability of the recombinant humanized collagen was first analyzed by treating at different temperatures,then its skincare efficacy based on in vitro level was evaluated by detecting the inhibition rate of elastase,the inhibition rate of collagenase,the protein content of type I collagen in human fibroblasts,the inhibition of reactive oxygen species(ROS)with human keratinocytes,and the effects of the recombinant humanized collagen on the expression of hyaluronic acid(HA),filaggrin(FLG)and transglutaminase 1(TGM1)in keratinocytes.The results showed that recombinant humanized collagen was able to maintain stability at temperatures below 70℃.With regard to its skincare efficacy,recombinant humanized collagen could inhibit elastase and collagenase activities and promote the increase of type I collagen content in human fibroblasts.It also showed good inhibition of ROS in keratinocytes in vitro and could increase the expression of HA,FLG,and TGM1 in keratinocytes.In short,the recombinant humanized collagen exhibited a favourable skin care effect in vitro level.This study proved that it has potential firming,anti-wrinkle,moisturizing,and repairing efficacy,and is a valuable cosmetic raw material.
基金Supported by the National Natural Science Foundation of China(32172521)。
文摘As an important wild blueberry resource,Vaccinium uliginosum has attracted more and more attention.At present,the wild resources are under destruction.The conservation of wild Vaccinium uliginosum resources is imminent.However,there are few researches on the protection and preservation of its germplasm resources.In vitro preservation is an important method for germplasm conservation.In this study,one strain of wild Vaccinium uliginosum was used as material.The effects of temperature(25℃,15℃,10℃,or 0℃),media(WPM,1/2WPM or 1/3WPM),medium supplements(sorbitol or mannose),and photoperiod(8,10,12,or 14 h•d^(-1))on the growth,survival rate and rejuvenation rate of the plantlets were studied.The physiological changes of plantlets during preservation were analyzed.Methylation-sensitive amplified polymorphism(MSAP)analysis of genomic DNA methylation of plantlets was carried out to explore the genetic stability of the plantlets after preservation.The research results provided a theoretical basis for the germplasm preservation of Vaccinium uliginosum.
文摘The in vitro maturation, fertilization and development of bovine ovary oocytes in two differentcultural systems A and B were studied under the conditions of 38.5℃,5%CO2,95%air and 100%humidity.The maturation rates were 94.5%and 91.3%,respectively,and the difference wasextremely significant.Frozen semen were thawed and sperm were capacitated with three kinds ofcapacitation agents for fertilization.The pronucleus rates were 76%,65%-68%and 62%respectively.The rates of embryos developed to morula and blastocyst were 19%,16% and 17%respectively.The developmental rates of embryos cocultured with bovine oviductal epithelium cellsand bovine granulosa cells were 25% and 23.4% respectively,with no significant difference. Freshembryos were transplanted into 15 recipicns,and three of them were pregnant and calves were bornin 1990 and 1991.The pregnant rate was 20%.The emryos developed faster before 8-cell stage andslower after 8-cell stage,in vitro than in vivo.
基金Supposed by the National Natural Science Foundation of China(32072749)。
文摘The antioxidant effects of quercetin were studied in vitro and in vivo.In vitro,vitamin C was used as a positive control to evaluate the antioxidant capacity of quercetin in three aspects:scavenging free radicals,protecting biological macromolecules and the total reducing power.In vivo,a total of 240 AA broilers(1-day age)with similar body weight were randomly divided into four groups with six replicates in each group,and 10 broilers in each replicate.The four groups were fed with corn-soybean basal diet supplemented with 0.00%,0.02%,0.04%and 0.06%quercetin to study its effects on antioxidant indexes of AA broilers,and to explore the optimal dose of quercetin as a dietary additive.The results showed that quercetin scavenged superoxide anion,hydroxyl radical and 1,1-diphenyl-2-picrylhydrazyl(DPPH)in vitro,the scavenging effects of quercetin on O_(2)-and•OH first increased and then decreased with the increase of the concentrations(P<0.01),and its maximum scavenging effect was observed at concentrations of 40 and 300 mg·L^(-1).The scavenging effects of quercetin on DPPH was increased constantly with increasing concentrations.The scavenging effect of quercetin on three free radicals was DPPH>•OH>O_(2)-.The inhibition of vitelline lipoprotein peroxidation by quercetin was increased with increasing concentrations(P<0.01)and the inhibitory effect was higher than that of vitamin C.The inhibition of red blood cell hemolysis by quercetin was increased with increasing concentrations at 0.05-1.25 mg·L^(-1)(P<0.01);however,the inhibition tended to decrease when the concentration was too high(31.25 mg·L^(-1)),and the inhibitory effect was higher than that of vitamin C.The inhibition of mitochondrial expansion by quercetin was increased with increasing concentrations,according to the degree of mitochondrial expansion at 60 min,the integrity of mitochondria in the experimental groups was significantly higher than that in the model group(P<0.01).The total reducing power of quercetin was increased with increasing concentrations(P<0.01);however,the total reducing power was less than that of vitamin C.In vivo,malondialdehyde(MDA)and nitric oxide(NO)were significantly decreased with increasing quercetin(P<0.01).Quercetin supplementation had no effect on the content of lipid peroxidation(LPO)in livers(P>0.05);however,superoxide dismutase(SOD)activity was significantly increased,whereas glutathione peroxidase(GSH-Px)and catalase(CAT)activities were significantly decreased in livers with increasing quercetin(P<0.05).These results suggested that quercetin exhibited strong antioxidant effects in vitro and in vivo.
基金Supported by Fund of Heilongjiang Acadamy of Agricultural Science
文摘Three methods were adopted in culture spermatogoniums of newly born calf in vitro,such as enzymatic digestion and percoll density gradient centrifugation（MethodⅠ）,tubular fragments culture（MethodⅡ）and tissue culture（MethodⅢ）,and cultural behaviors of cells were observed.The results showed that typical spermatogonium colonies appeard at 144 h of culture by enzymatic digestion-percoll density gradient centrifugation method and tubular fragments culture method,2.5%FBS kept the characteristics of spermatogonium stem cell better than others,produced more mass clones,and FBS of more than 2.5%concentration benefited spermatogonium differentiation and the number of colonies was significantly affected by FBS concentration.After 1 week of culture in method Ⅲ,the diameter of lumens and quantity of sertoli’s cells in tubal wall increased obviously,lumen of seminiferous tubules appeared.Sertoli’s cells kept constant and the number of spermatogoniums decreased obviously after 2 weeks of culture.
基金Project(2012zzts068) supported by the Fundamental Research Funds for the Central Universities of Central South University,ChinaProject(2010fj3091) supported by the Open Funding of State Key Laboratory of Powder Metallurgy and Science&Technology Foundation,China
文摘Mg-6%Zn-10%β-Ca3(PO4)2 composite was prepared through powder metallurgy methods with different chitosan coatings on its surface. The properties of the chitosan coatings on the surface of Mg-6%Zn-10%β-Ca3(PO4)2 composite, such as the adhesion ability, the corrosion behavior and the cytotoxicity properties, were investigated, and the microstructure of the chitosan coating was observed by scanning electron microscope(SEM). The results show that chitosan coating improves the corrosion resistance of the magnesium composite specimens significantly. Mg-6%Zn-10%β-Ca3(PO4)2 composite specimens exhibit good corrosion resistance and low p H values in simulated body fluid(SBF) at 37 °C in the immersion test with 7-layer chitosan coating whose relative molecular mass is 30×104 Da. The cytotoxicity tests indicate that Mg-6%Zn-10%β-Ca3(PO4)2 with chitosan coating is nontoxic with a cytotoxicity grade of zero against L-929 cells, which is better than that of uncoated composites.
基金Natural Science Foundation of Hebei Province(H2022206456)Natural Sci⁃ence Foundation of Hebei Province(H2021206449)+1 种基金Undergraduate Innovative Experiment Program of Hebei Medical University(USIP2022173)Undergraduate Innovative Experiment Program of Hebei Medical University(USIP2023107)。
文摘OBJECTIVE To explore mecha⁃nisms of imperatorin on regulating P-glycoprotein(P-gp)in blood-brain barrier(BBB)based on net⁃work pharmacology combined with in vitro experi⁃ment.METHODS Drug targets were predicted using the Pharmapper and Swiss targets data⁃bases;disease targets were obtained through the Genecards database;intersections between drugs and disease targets were screened by Cytoscape software;the obtained core targets were used to construct protein-protein interaction(PPI)network,gene ontology(GO)functions,and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.The effects of imperatorin(20,50,100μmol·L^(-1))on P-gp activity were monitored in hCMEC/D3 in vitro BBB model,and the effects of imperatorin on the expression of target proteins were verified using Western blot method.RESULTS 55 drug targets and 3102 disease targets were obtained from the network pharmacology screening,and 37 core targets were obtained after the combination.Enrichment analysis showed that core targets were closely related to chemical synaptic trans⁃mission regulation,neurotransmitter receptor activity,protein kinase regulation activity,G proteincoupled receptor signaling pathway,neural active ligand receptor interaction pathway,PI3K-Akt sig⁃naling pathway,VEGF signaling pathway,etc..In vitro experimental validation suggested that all tested concentration groups of imperatorin signifi⁃cantly reduced the activity and expression of P-gp,which were achieved by significantly downregu⁃lating the phosphorylation levels of PI3K and Akt,and repressing the expression of VEGFR2 pro⁃tein.CONCLUSION Network pharmacology was used to predict the core targets and signaling pathways of imperatorin on regulating P-gp in BBB and relevant validation was conducted through in vitro experiments,providing a refer⁃ence basis for further exploration of the mecha⁃nisms of imperatorin on regulating P-gp in BBB.
基金The project supported by Young Scientist Funding from Beijing Natural Science Foundation(7154225)by Innovative Research Team in IMPLAD
文摘Influenza virus infection is a global public health issue.The effectiveness of antiviral agents for influenza has been limited by the emergence of drugresistant virus strains.Therefore,there is an urgent need to identify novel antiviral therapies.Our previous studies have found that Cryptoporus volvatus extract could potently inhibit influenza virus replication in vitro and in vivo.However,the effective component of Cryptoporus volvatus which mediated the antiviral activity hasn′t been identified.Here,we identified a novel anti-influenza molecule,cryptoporic acid E(CAE),from Cryptoporus volvatus.Our results showed that CAE had broad-spectrum anti-influenza activity against 2009 pandemic strain A/Beijing/07/2009(H1N1/09),seasonal strain A/Jiangxi/262/05(H3N2),mouse adapted strains A/WSN/33(H1N1)and A/PR8/34(H1N1).We further investigated the mode of CAE action,and found that CAE directlyattenuated influenza virus infectivity.Time-course-analysis indicated that CAE exerted its inhibition mainly at middle stage of the replication cycle of influenza virus.Subsequently,we confirmed that CAE blocked virus RNA replication and transcription in MDCK cells and CAE repressed influenza virus RNA polymerase activity.In addition,we found that CAE impaired influenza virus infectivity by directly targeting virus particles.Our data suggest that CAE is a major effective component of Cryptoporus volvatus and might be a potential candidate for the development of a new anti-influenza virus therapy.
基金Supported by the Scientific Research fund of the Guangxi Zhuang Autonomous Region(GXNYRKS201601)
文摘In this paper, several factors that affect the efficiency of in vitro adventitious bud regeneration and Agrobacterium tumefaciens-mediated transformation of F. vesca were studied. The results showed that F. vesca seeds germination rate was the highest while seeds were cultured in water, and the germination rate was the lowest while seeds were cultured on MS medium supplemented with hormone; the germination rates that seeds cultured on two and three layers filter paper were higher than that seeds cultured on four and five layers filter paper. In vitro adventitious regeneration efficiency was affected by different explants types. The significant difference was existed between petioles and leaves. When using the same type explants, in vitro adventitious buds regeneration rate and the average number of buds per explant between Ruegen （RE） and Yellow Wonder （YW） had no significant difference. RE to Agrobacterium tumefaciens was more sensitive than YW. Using seedling leaves of RE and YW as materials, an efficient Agrobacterium-mediated transformation system was developed. In this system, the concentration of bacteria was OD600=0.5, the explants were immersed in bacteria broth for 9 min, the co-cultured time was 2 days, and had no pre-cultured time. The percentage of explants with resistant buds of RE and YW was compared. The putative transformed plants were confirmed by PCR.
基金Supported by the Key Teacher Foundation of Education Offi ce of Heilongjiang Province(2000)
文摘To investigate the effects and mechanisms of the benzimidazole carbamate and benmidine drugs on Cysticerci cellulosae and choose effective drugs on Cysticerci cellulosae, the membrane metabolism of Cysticerci cellulosae in vitro was tested after three kinds of drugs which were used respectively. The indexes included the contents of lipids, the contents of SA and the changes of the membrane fluidity. The results showed that oxfendazole could inhibit the membrane metabolism of immature and mature Cysticerci cellulosae in vitro, and albendazole only inhibited the mature one, while thibendimidine neither acted on the immature nor mature one.
文摘Collected from the broiler cecal contents, bacteria were isolated and purified. These isolated and purified Lactobacilli were detected by acidity and bile salt tolerant test, then three better tolerant strains were screened out. These three strains were conducted to observe the morphous through microscope, perform Gram stain, and to identify their physiological and biochemical characteristics. The results showed that L1 was L.casei, L2 was L.gasseri and L3 was L.graminis. With Oxford Cup method, these three Lactobacilli were detected for their bacteriostasis. Results showed that L1, L2 and L3 strains had strong antibacterial activities to E. coli and Salmonella.
文摘The histological observation was experimentally conducted on in vitro cultured mouse embryonic myocardium cells and myocardiumoid cell mass. The mouse embryo tissue were cultured and regular pulsatile myocardiumoid tissue could be found. During in vitro culture, the myofilament bundles in the cell were gradually increasing and strongly connectted each other with embryonic age and there were loose muscle fibers initially and intercalated discs were close to each other. The lose myofilament bundles were developed in muscle fibers with age and the distance between intercalated discs was enlarged. There were myofilamentoid structure in inactive cells and filament peripherily.
文摘Cyclamen leaves and petioles explants were cultured on MS media supplemented with different concentrations of 2, 4-dichlorophenoxyacetic acid (2, 4-D) or 1-naphthaleneacetic acid (NAA) to induce callus. The effect of 2, 4-D on shoot regeneration was also studied. Either in media containing 2, 4-D or in media containing NAA, callus was observed, but the quality or quantity of callus induced by 2, 4-D or NAA were different. The callus induced by 2, 4-D was white, compact and having powerful multiplication capacity. The callus was inclined to browning then was poorly organogenetic. While the callus induced by NAA was yellowish in appearance. It was pultaceous and proliferated bradytelicly. The callus usually can give rise to many shoots. But the frequency of inducing callus of 2, 4-D is higher than that of NAA. The regenerative plantlets derived from the callus respectively induced by 2, 4-D or NAA were transferred into rooting medium. The frequency of rooting were no difference.
基金Supported by the Special Fund for Agro-scientific Research in the Public Interest(201503134)。
文摘Batch cultures of mixed rumen micro-organisms were conducted to evaluate the effects of encapsulated yeast(+EY)and encapsulated enzyme(+EE)using plant proteins(barley and oats grain)on rumen fermentation in vitro,investigate the abilities of encapsulated yeast and encapsulated enzyme to prevent rumen digestion in vitro.Treatments of the study were the control,+EY,+EE products(3.33 mg·mL^(-1) of the incubation medium),unencapsulated yeast(-EY)and enzyme(-EE)products(0.17 and 0.17μL·mL^(-1) of the incubation medium,respectively).+EY group increased dry matter disappearance(DMD,P<0.01)and the total volatile fatty acids(TVFA,P<0.01)at 3 h of the incubation compared with the control,regardless of encapsulation of yeast.Gas production(GP)of+EY group was higher(P=0.05,29.94 mL·mL^(-1) organic matter,OM)than that of the control(25.08 mL·g^(-1) OM)at 3 h of the incubation.Supplementation+EY increased DMD(P=0.04,0.394 vs 0.352,respectively)and acetic proportion(P=0.04,52.6 vs 49.8 mol•100 mL^(-1),respectively)at 6 h of the incubation and increased A:P ratio(P<0.01,3.11 and 2.86,respectively)at 24 h of the incubation,as compared to unencapsulation of yeast.Supplementation of enzyme had higher(P≤0.04)GP,DMD and TVFA at 3 and 6 h of the incubation compared with the control,regardless of encapsulation.Moreover,the addition of+EE produced greater GP at 6(P<0.01,92.35 vs 78.21 mL·g^(-1) OM,respectively),12(218.47 vs 159.18 mL·g^(-1) OM)and 24 h(380.97 vs 297.78 mL·g^(-1) OM,respectively)of the incubation,higher DMD(0.347 vs 0.313,respectively)at 3 h of the incubation as compared to-EE group.The study showed that the encapsulation might protect part of yeast and enzyme from releasing to the rumen throughout the digestion in vitro,resulting in higher or no difference of rumen fermentation parameters compared with unencapsulated groups at any incubation times.In comparison with-EY and-EE,the higher rumen fermentation parameters at the early incubation time were observed,which could be attributed to the higher concentration of yeast or enzyme.However,regardless of the encapsulation,the results indicated that both yeast and enzyme only improved the speed rather than the extent of rumen fermentation in vitro.
基金supported by a predoctoral fellowship from the National Institutes of Health and a research grant from the National Science Foundation
文摘Introduction Cancer is an attractive target of gene therapy and currently represents the disease in most clinical trials[1]. Strategies for cancer gene therapy include: (1) stimulation of immune responses to tumor cells,(2) delivery of specific enzymes
文摘The present investigation was undertaken to find out the best medium composition for medium-term preservation of disease free potato shoot tips in in vitro system.Thirteen potato genotypes and fourteen treatments were taken under consideration for the present experiment.Among fourteen treatments,mannitol and sorbitol containing media proved to be the best for medium-term preservation of potato shoot tips.High concentration of mannitol delayed root formation.
文摘Biomaterials for restoration or replacement of diseased tissues may have any origin.The major characteristic for biomaterials is biocompatibility.All biomaterials,used in medicine (dentistry,in particular),interreact with the organism tissues.And the changes occur both in the materials and the organism tissues.It is considered that there are no "inert biomaterials." The number of allergic diseases and complications is constantly growing all over the world,taking an important place in the structure of infectious and noninfectious pathology[1].Pollen,household,epidermal,and food-borne allergens,and haptens are the most frequent sources of sensibilization.
