Alzheimer's disease (AD) is a progressive neurodegenerative disorder and is the most prominent cause of dementia.In 2019,over 57.4million people were living with AD and other dementia subtypes,a number which is ex...Alzheimer's disease (AD) is a progressive neurodegenerative disorder and is the most prominent cause of dementia.In 2019,over 57.4million people were living with AD and other dementia subtypes,a number which is expected to increase to over 152.8 million in the next 25years.This ever-increasing burden has resulted in AD and other neurodegenerative diseases rising to one of the top 10 causes of death globally (O'Connell et al.,2024).展开更多
In Alzheimer’s disease,microglial phagocytosis is engaged in the pathogenesis as it clears abnormal protein accumulations,debris,and apoptotic cells in the early stages of Alzheimer’s disease,but fuels neuroinflamma...In Alzheimer’s disease,microglial phagocytosis is engaged in the pathogenesis as it clears abnormal protein accumulations,debris,and apoptotic cells in the early stages of Alzheimer’s disease,but fuels neuroinflammation and accelerates disease progression in later stages.In vivo parabiosis experiments in aged animals have demonstrated that blood-born factors modulate synaptic plasticity,neurogenesis,and microglial responses.We hypothesize that peripheral factors can modulate microglial function and thereby possibly influence Alzheimer’s disease pathology.The objective of this study is to investigate the effects of Alzheimer’s disease serum on microglial phagocytosis.Here,we use an immortalized human microglial cell line in an in vitro parabiosis assay to investigate the impact of the serum from individuals diagnosed with Alzheimer’s disease(n=30)and age-matched controls(n=30)(PRODEM study)on microglial phagocytosis.Exposure to Alzheimer’s disease serum increased microglial phagocytic uptake of pH-sensitive fluorescent particles and downregulated expression of the lysosomal master regulator transcription factor EB(TFEB)and of ATPase H^(+)transporting lysosomal V1 subunit B2(ATP6V1B2),a component of the vacuolar ATPase.To identify serum components that may relate to changes in phagocytosis,serum samples of the Three-City Study(3C Study)were used.In the 3C Study,blood samples were collected up to 12 years before the onset of cognitive decline or dementia and their serum metabolome is well-defined.Microglia exposed to the serum of future Alzheimer’s disease patients from the 3C Study displayed an increased phagocytic uptake compared with the serum of matched controls,depending on the presence of the apolipoprotein Eε4 allele in the Alzheimer’s disease patients.Furthermore,microglial phagocytosis correlated inversely with serum levels of the omega-3 fatty acid eicosapentaenoic acid.We confirmed this inverse correlation between eicosapentaenoic acid and phagocytosis in the serum samples of the PRODEM cohort.In addition,in vitro testing of eicosapentaenoic acid on microglial phagocytosis showed a concentration-dependent decrease in phagocytic uptake.In conclusion,following incubation with Alzheimer’s disease blood serum,we observed increased microglial phagocytic uptake and the downregulation of TFEB and ATP6V1B2,possibly indicating lysosomal dysfunction.Furthermore,microglial phagocytosis was inversely correlated with serum eicosapentaenoic acid levels,suggesting an important role for dietary eicosapentaenoic acid in microglial function.展开更多
This study aimed to optimization of the in vitro fertilization system in Cỏ goat oocytes to achieve the maximum possible blastocyst development rate. In Experiment 1, we assessed the effects of IVF media on the in vit...This study aimed to optimization of the in vitro fertilization system in Cỏ goat oocytes to achieve the maximum possible blastocyst development rate. In Experiment 1, we assessed the effects of IVF media on the in vitro fertilization of Cỏ goat oocytes. There was no significant difference in the cleavage, blastocyst, or hatching rates between TALP-Fert and BO-IVF media. Experiment 2 was performed to assess the concentration of sperm in the in vitro fertilization of Cỏ goat oocytes. The matured Cỏ goat oocytes were fertilized in BO-IVF for four sperm concentrations: 5 × 105, 1 × 106, 2 × 106 and 3 × 106 sperm/ml. The blastocyst rate of 2 × 106 sperm/ml and 3 × 106 sperm/ml groups was higher than that of 5 × 105 sperm/ml and 1 × 106 sperm/ml groups (P Experiment 3 was performed to assess the IVF duration on the in vitro fertilization of Cỏ goat oocytes. The matured Cỏ goat oocytes were fertilized in BO-IVF with sperm concentration of 3 × 106 sperm/ml for 18, 20, 22 and 24 h. The cleavage, blastocyst, and hatching blastocyst rates of 18 h group were lower than those of 20, 22 and 24 h groups (P 0.05). In conclusion, the matured Cỏ goat oocytes were fertilized in BO-IVF with sperm concentration of 3 × 106 sperm/ml for 20 hours, which is suitable for the in vitro Cỏ goat embryo production.展开更多
Objective:This study aims to investigate the energy conduction mechanism of holmium laser lithotripsy based on the Gemini platform and validate the optimal lithotripsy parameters under this mechanism using an in vitro...Objective:This study aims to investigate the energy conduction mechanism of holmium laser lithotripsy based on the Gemini platform and validate the optimal lithotripsy parameters under this mechanism using an in vitro ureteral model.Methods:The Lumenis VersaPulse PowerSuite dual-wavelength laser system was employed,with a holmium laser wavelength of 2.1μm,power output of 0-80 W,pulse energy ranging from 0.2 to 3.5 J,and pulse frequency from 5 to 40 Hz.An in vitro thermal effect model of the holmium laser was established to simulate temperature changes in urinary stones under varying power levels,energy ranges,pulse frequencies,and irrigation rates.High-speed cameras and thermal imaging devices were used to observe and record the temperature of the stone and surrounding tissues during laser irradiation.Finite element analysis was integrated to simulate the energy conduction pathways of the holmium laser within the stone and ureteral wall,assessing whether localized high-temperature regions exceeded the thermal injury threshold.Results:Temperature in the stone fragmentation mode decreased significantly with increasing distance from the basket and higher irrigation rates,thereby reducing the risk of thermal injury.In fragmentation mode,both 276μm and 500μm core-diameter fibers generated significantly higher temperatures at the basket site compared to the free-running mode(P<0.05).No thermal injury was observed at irrigation rates≥15 ml/min across all parameter combinations;at 10 ml/min,only high-energy settings(3.5 J)approached the thermal injury threshold(50.3℃);at≤5 ml/min,high-energy settings(>1.5 J)consistently exceeded the danger threshold.The safest parameter combination was 0.5 J+5 Hz(at any irrigation rate).Within 5 min,temperatures at the stone surface-below,middle,and above the basket-reached 151.0℃±1.0℃,62.5℃±0.2℃,and 46.8℃±0.2℃,respectively,whereas the maximum temperature on the ureteral wall surface was only 49.5℃.The FEA results closely matched experimental observations,indicating that with 0.5 J+5 Hz(at any flow rate),laser energy is primarily confined within the stone,generating localized high-temperature zones(80℃-90℃)at the stone-ureter wall interface,which promotes stone fragmentation while effectively minimizing collateral tissue damage.Conclusion:The holmium laser energy conduction mechanism based on the Gemini platform is predominantly concentrated within the stone,exhibiting excellent directionality and depth penetration,enabling efficient lithotripsy while minimizing thermal damage to surrounding tissues.展开更多
Infertility,defined as the inability to achieve a clinical pregnancy after 1 year of unprotected and regular sexual intercourse,affects approximately 8%–12%of couples worldwide during their childbearing years.Assiste...Infertility,defined as the inability to achieve a clinical pregnancy after 1 year of unprotected and regular sexual intercourse,affects approximately 8%–12%of couples worldwide during their childbearing years.Assisted reproductive technology(ART)offers interventions to facilitate conception in couples with infertility.Since the inception of traditional in vitro fertilization(IVF)in 1978,ART has facilitated the birth of millions of infants.Intracytoplasmic sperm injection(ICSI)is an effective technique for couples ineligible for traditional IVF.Individual female factors,such as age and endometrial thickness,have been shown to affect embryo development during the IVF process,resulting in unfavorable pregnancy outcomes[1-3].However,the influence of environmental and male factors on early reproductive outcomes cannot be ignored.As a novel form of environmental pollution,light at night(LAN)has intensified with the rapid pace of urbanization,potentially leading to reproductive health problems in both women and men.However,to our knowledge,no study has explored the effects of LAN exposure on the early reproductive outcomes of IVF.Furthermore,the normal development of embryos relies on the contributions of both partners,and the influence of male factors on the early reproductive outcomes of IVF should not be overlooked.展开更多
Correction to:Neurosci.Bull.December,2016,32(6):577–584.https://doi.org/10.1007/s12264-016-0068-z In this article,in Fig 5A,the picture of the Vector+Nogo-66 group was incorrect and should have appeared as shown below.
This study employed a microwave-ultrasonic-hydrothermal multifield coupling method to synthesize nanoβ-Tricalcium phosphate(β-TCP)powder,systematically evaluating the impact of various parameters,including reaction ...This study employed a microwave-ultrasonic-hydrothermal multifield coupling method to synthesize nanoβ-Tricalcium phosphate(β-TCP)powder,systematically evaluating the impact of various parameters,including reaction temperature,time,sintering temperature,reactant types and concentrations,and graphene oxide(GO)concentration,on the physicochemical properties of the nanoβ-TCP powder.The synthesized powder was characterized using X-ray diffraction(XRD),Fourier-transform infrared spectroscopy(FTIR),scanning electron microscopy(SEM),high-resolution transmission electron microscopy(HRTEM),inductively coupled plasma optical emission spectroscopy(ICP-OES),and thermogravimetric-differential scanning calorimetry(TG-DSC).The experimental results indicate that the optimal synthesis conditions are achieved with a 0.6 mol/L Ca(NO3)2·4H2O solution and a 0.4 mol/L(NH_(4))_(2)HPO_(4)solution at a reaction temperature of 35℃for 40 minutes,followed by sintering at 720℃for 2 hours with 1×10^(-1)g/L GO.The preparedβ-TCP powder exhibits high crystallinity,a pure phase,good dispersibility,no significant aggregation,and uniform particle size of 59.75±12.84 nm.In vitro cytotoxicity tests show excellent biocompatibility and no cytotoxic effects on bone marrow stromal cells(BMSCs)even at concentrations up to 0.8 mg/mL.Furthermore,results from live-dead staining and nuclear membrane staining of cells co-cultured with the material demonstrate that theβ-TCP can promote the proliferation and differentiation of BMSCs to a certain extent,highlighting its potential as a safe and effective material for bone tissue engineering.展开更多
The global demand for in vitro respiratory airway models has surged due to the coronavirus disease 2019(COVID-19)pandemic.Current state-of-the-art models use polymer membranes to separate epithelial cells from other c...The global demand for in vitro respiratory airway models has surged due to the coronavirus disease 2019(COVID-19)pandemic.Current state-of-the-art models use polymer membranes to separate epithelial cells from other cell types,creating a nonphysiological barrier.In this study,we applied three-dimensional(3D)printing and bioprinting to develop an in vitro model where endothelial and epithelial cells were in direct contact,mimicking their natural arrangement.This proof-ofconcept model includes a culture chamber,with an endothelial bioink printed and perfused through an epithelial channel.In silico simulations of the air velocity within the channel revealed shear stress values ranging from 0.13 to 0.39 Pa,aligning with the desired in vivo shear stress observed in the bronchi regions(0.1–0.4 Pa).Biomechanical movements during resting breathing were mimicked by incorporating a textile mesh positioned away from the cell–cell interface.The epithelial channel demonstrated a capacity for compression and expansion of up to−14.7%and+6.4%,respectively.Microscopic images showed that the epithelial cells formed a uniform monolayer within the lumen of the channel close to the bioprinted endothelial cells.Our novel model offers a valuable tool for future research into respiratory diseases and potential treatments under conditions closely mimicking those in the lung.展开更多
Objective:This review aims to investigate and establish potential in vitro and in vivo models for evaluating the anti-urolithiatic activity of therapeutic agents,exploring experimental approaches that can reliably sim...Objective:This review aims to investigate and establish potential in vitro and in vivo models for evaluating the anti-urolithiatic activity of therapeutic agents,exploring experimental approaches that can reliably simulate human stone formation and effectively assess the efficacy of anti-urolithiatic compounds.Methods:Multiple in vitro and in vivo approaches were explored.In vitro methods included the estimation of calcium oxalate by titrimetry,nucleation assays,aggregation assays,turbidimetric assays,and electron microscopy studies.Artificial stone models such as BegoStone and Ultracal 30 were fabricated to mimic the physicochemical characteristics of urinary calculi.In vivo models included ethylene glycol-induced,calcium oxalate/ammonium oxalate-induced,diet-induced,and infection-related models in rodents.Additionally,genetically modified animal models such as TRPV5 knockout,CLDN14 knockout,AGXT knockout,and URAT1 overexpression mice were discussed to study molecular pathways of urolithiasis.Parameters such as urinary oxalate,calcium levels,and histopathological evaluation of kidney tissues were used to validate stone formation and dissolution processes.Results:In vitro models effectively demonstrate the processes of crystal nucleation,aggregation,and inhibition,allowing quantitative assessment of potential anti-urolithiatic activity.Electron microscopy provides detailed insights into crystal morphology and ultrastructural alterations.Artificial stones fabricated using BegoStone and Ultracal 30 closely replicate natural calculi hardness and composition,making them suitable for lithotripsy and dissolution studies.In vivo models successfully mimic human urolithiasis pathophysiology,particularly the ethylene glycol-induced rat model,which shows reproducible calcium oxalate crystal deposition in renal tissues.The application of genetic models highlights the role of specific transporters and enzymes in calcium and oxalate homeostasis.Conclusion:A combination of in vitro and in vivo experimental models provides a comprehensive platform for evaluating the anti-urolithiatic potential of therapeutic agents.The integration of biochemical,morphological,and genetic analyses enhances the understanding of stone pathogenesis and development of novel anti-urolithiatic therapies.展开更多
As prepubertal boys do not yet produce spermatozoa,they cannot rely on sperm cryopreservation for fertility preservation before gonadotoxic therapy,such as high-dose alkylating agents or radiotherapy in the case of ch...As prepubertal boys do not yet produce spermatozoa,they cannot rely on sperm cryopreservation for fertility preservation before gonadotoxic therapy,such as high-dose alkylating agents or radiotherapy in the case of childhood cancers.According to the current guidelines,cryopreservation of testicular biopsies containing spermatogonial stem cells(SSCs)may be proposed to high-risk patients for potential later therapeutic use to fulfill the patients’wish for a biological child.One promising technique for human in vitro spermatogenesis and in vitro propagation of human SSCs is microfluidic(MF)culture,in which cells or tissues are subjected to a continuous flow of medium.This provides exact control over such parameters as nutrient content and gradients,as well as the removal of waste metabolites.While MF has been shown to maintain tissues and cell populations of organs for longer than conventional in vitro culture techniques,it has not been widely used for testicular in vitro culture.MF could advance human testicular in vitro culture and is also applicable to reprotoxicity studies.This review summarizes the findings and achievements of testis-on-chip(ToC)setups to date and discusses the benefits and limitations of these for spermatogenesis in vitro and toxicity assessment.展开更多
Rapid and sensitive detection of targeted biomarkers in trace samples is of great significance for early in vitro diagnosis of diseases.Microfluidic technology has competitive advantages in this field due to its low c...Rapid and sensitive detection of targeted biomarkers in trace samples is of great significance for early in vitro diagnosis of diseases.Microfluidic technology has competitive advantages in this field due to its low cost,high efficiency,and high portability;however,the analysis of results tends to rely on bulky and sophisticated instruments,and this limits its applications.In this work,we developed a Raspberry Pi camera-based biomarker detection device based on microfluidic technology and digital image colorimetry.For highly sensitive biomarker detection on microfluidic chips,we propose a three-step signal-amplification colorimetric detection strategy consisting of:(1)the release of Ag^(+)ions from silver nanoparticles,(2)Ag^(+)-inhibited urea hydrolysis colorimetry,and(3)microscopic lens magnification.For efficient evaluation of results,we employed an RGB image-processing system to quantitatively analyze color images captured by the Raspberry Pi camera.Further,we tested the functionality of the device with procalcitonin(PCT)in phosphate-buffered saline,plasma,and serum to simulate clinical situations.We determined the limit of detection as 1 ng/ml,and a good linear relationship was established between PCT concentration and color intensity within the detection range 1–10 ng/ml.Importantly,only a relatively short detection time(40 min)was required in all three environments.The results demonstrate the great potential of this device for biomarker detection and facilitating biomedical research.展开更多
The olive oil fermentation filtrate was prepared by microbial fermentation technology,and the fermentation process was optimized by orthogonal test to screen out the optimal fermentation conditions.At the same time,th...The olive oil fermentation filtrate was prepared by microbial fermentation technology,and the fermentation process was optimized by orthogonal test to screen out the optimal fermentation conditions.At the same time,the effect of olive oil fermentation filtrate to promote the secretion of type I collagen (COL-I) and type VII collagen (COL-VII) by fibroblasts (HFF-1),the inhibitory effect on the secretion of alpha tumor necrosis factor (TNF-α) and interleukin-6 (IL-6) by lipopolysaccharide induced macrophages (RAW264.7),and the inhibitory effect on the induction of reactive oxygen species (ROS) production by immortalized human keratinocytes (HaCaT) were studied through in vitro cell testing methods.The results showed that the optimal fermentation conditions of olive oil fermentation filtrate were as follows,inoculation amount was 3%,inducer amount was 2%,fermentation time was 48 h.Its efficacy can significantly promote the synthesis of COL-I and COL-VII,and significantly inhibit the content of inflammatory factors TNF-α and IL-6 as well as ROS content,it has multiple effects of firming,anti-wrinkle,soothing and antioxidant,providing scientific and effective data support for the development of cosmetic raw materials.展开更多
Considerable levels of methylmercury(MeHg)have been found rice-based infant cereals as a result of MeHg transfer from the rice as a raw material to the products,hence consumption of rice products may pose a potential ...Considerable levels of methylmercury(MeHg)have been found rice-based infant cereals as a result of MeHg transfer from the rice as a raw material to the products,hence consumption of rice products may pose a potential health risk to infants who may receive cereals as the major diets and are susceptible to toxicity of MeHg.Determination of bioaccessibility of MeHg would provide a more accurate assessment of MeHg exposure through consumption of rice-based cereals,yet this information remains lacking.Further,the re-adsorption of methylmercury on the residual food will affect the accurate assessment of its bioaccessibility.Our goals in this work were 1)to determine the bioaccessibility of MeHg in infant rice cereals commonly available on the market by using a typical in vitro artificial gastrointestinal digestionmodel and 2)to evaluate the effects of MeHg re-adsorption on the in vitro assessment of MeHg bioaccessibility.The determined bioaccessibility of MeHg in the studied rice cereals after the standard dual-step(using gastric followed by intestinal juice)protocol ranged from 25%to 74%,a wide range comparable to that of fish samples observed here and in previous studies.The surprisingly higher bioaccessibility of MeHg in fish and rice cereals after the gastric step only,in comparison to after the complete two-step gastrointestinal digestion,suggests the re-adsorption of MeHg on the residual food.Separate experiments with spiked MeHg standards confirmed that the MeHg released by the acetic gastric juice was re-adsorbed on the residues during the intestinal step at neutral pH.This study provided first-hand data on the bioaccessibility of MeHg in infant rice cereals and methodological implications on using in vitro digestion to evaluate the bioaccessibility of MeHg and metal contaminants in general.展开更多
Edible mushroom polysaccharides demonstrate a range of bioactivities and have been investigated as potential adjuvant therapies for cancer treatment.Our prior research found Lactariks hatsudake Tanaka polysaccharide(L...Edible mushroom polysaccharides demonstrate a range of bioactivities and have been investigated as potential adjuvant therapies for cancer treatment.Our prior research found Lactariks hatsudake Tanaka polysaccharide(LHP)effective against hepatocellular carcinoma,but the mechanisms remain unclear.This study aims to clarify the anti-cancer molecular mechanisms by examining the effects on specific signaling pathways through in vivo and in vitro analyses.Animal studies showed that LHP significantly inhibited the growth of transplanted liver cancer without toxicity.Histological and immunohistochemical analyses revealed that LHP increased nuclear cohesion in tumor cells and altered the expression of key regulatory proteins such as protein kinase B(Akt),nuclear factorκB(NFκB),caspase-3,p21,and p53.These findings suggest that LHP’s anticancer effects involve promoting apoptosis and disrupting the cell cycle.At the cellular level,assays such as 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS),flow cytometry,and transwell invasion confirmed that LHP inhibited cancer cell proliferation and induced apoptosis.Polymerase chain reaction and Western blot assays revealed that LHP intervention reduced cyclin-dependent kinase 4 activity by upregulating p53 and p21,leading to cell cycle arrest and inhibited cell proliferation.LHP also promoted apoptosis by altering Bcl-2-associated X protein,B-cell lymphoma 2,and cleaved caspase-3 levels.Moreover,LHP could affect the Akt,NF-κB,and mitogen-activated protein kinase signaling pathways to exert anticancer effects.These findings underscore the potential of LHP as a novel and multifaceted therapeutic agent.Furthermore,the elucidation of LHP-related mechanisms offers a crucial theoretical foundation for developing innovative and effective treatments for hepatocellular carcinoma.Additionally,this supports its potential development as a functional food or adjuvant therapy for cancer,which holds significant implications and application prospects in both the food and medical fields.展开更多
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
Pimpinella anisum,commonly known as anise,is generally used in both folk medicine and the culinary world.In traditional medicine,it is valued for its digestive,respiratory,and antispasmodic properties.This study aims ...Pimpinella anisum,commonly known as anise,is generally used in both folk medicine and the culinary world.In traditional medicine,it is valued for its digestive,respiratory,and antispasmodic properties.This study aims to examine the volatile compounds and antibacterial effect of P.anisum essential oil(PAEO)as well as for the first time its genotoxicity employing both in vitro and computational approaches.Gas chromatography-mass spectrometry(GC-MS)analysis identified anethole as the principal compound,which comprises 92.47%of PAEO.PAEO was tested for its potential antibacterial properties against Bacillus subtilis ATCC 6633,Listeria innocua ATCC 33090,Staphylococcus aureus ATCC 29213,Klebsiella aerogenes ATCC 13048,and a clinical strain of Salmonella enterica serotype Typhi.PAEO displayed noteworthy antibacterial action toward all tested bacteria,especially Staphylococcus aureus,with an inhibition zone of 21.43±0.87 mm,as determined by the disc-diffusion test.Varied between 0.0625%and 2%v/v,while the MBC values ranged from 0.125%to 8%v/v,reflecting the strength of the tested EO.The MBC/MIC ratios indicated the bactericidal nature of PAEO.The results of molecular docking revealed strong binding interactions between key PAEO molecules and microbial target proteins.ADMET(Absorption,Distribution,Metabolism,Excretion,and Toxicity)analysis confirmed favorable pharmacokinetic properties,indicating its potential as a safe therapeutic agent.Additionally,genotoxicity was assessed using the comet assay,which demonstrated minimal genotoxic risk,affirming the oil’s safety.These results highlight the promising antimicrobial properties of PAEO and its possible use as an active agent in the pharmacy,food,and cosmetic sectors.展开更多
Phalaenopsis orchids are economically important ornamental crops;however,their commercial micropropagation is often limited by poor rooting efficiency and inconsistent growth.In this study,we investigated the effects ...Phalaenopsis orchids are economically important ornamental crops;however,their commercial micropropagation is often limited by poor rooting efficiency and inconsistent growth.In this study,we investigated the effects of silver nanoparticles(Ag-NPs)on the in vitro regeneration and growth of Phalaenopsis cultivar 611B to determine the optimal concentration of Ag-NPs for improved micropropagation outcomes.Shoot tip explants(2–3 mm)—derived from protocorm-like bodies were cultured on a regeneration medium containing Hyponex(20:20:20 and 6.5:6.5:19),18 g/L sugar,2 g/L peptone,0.8 g/L activated charcoal,12.5 g/L potato extract,50 mL/L apple juice,and 10 mg/L 6-benzylaminopurine(6-BA),with varying concentrations of Ag-NPs(0,0.5,1.0,2.0,and 2.5 mg/L).After 10–12 weeks,shoot and root formation,plant height,fresh weight,leaf number,and chlorophyll contents were evaluated.At 1.0 mg/L Ag-NPs,shoot regeneration(5.4 vs.2.9 shoots per explant),root induction(2.1 vs.1.4 roots per explant),and shoot formation frequency(100%vs.55%)were significantly higher than the control(0 mg/L).Fresh weight(592.4 mg)and leaf number(9.7)also showed notable increases at this concentration.Although chlorophyll a and b levels peaked at 2.0 mg/L,the difference from 1.0 mg/L was not statistically significant.These results suggest that 1.0mg/LAg-NPs is the optimal concentration for enhancing shoot and root development and improving overall plantlet quality in Phalaenopsis.The findings highlight the potential of nanomaterials to improve the efficiency of orchid tissue culture systems.展开更多
The gastrointestinal(GI)tract is essential for digestion,absorption,excretion,and protection,supported by a diverse microbial ecosystem.Traditional in-vitro models often fall short in capturing the physiological compl...The gastrointestinal(GI)tract is essential for digestion,absorption,excretion,and protection,supported by a diverse microbial ecosystem.Traditional in-vitro models often fall short in capturing the physiological complexity of the GI tract,limiting their translational applications.A comprehensive approach is needed to bridge the gap between simple cell cultures and more complex systems used in translational research.This review explores the limitations of conventional twodimensional cell cultures and emphasizes the emerging use of three-dimensional and microfluidic systems that better replicate the GI tract’s structure and functions.It highlights the importance of incorporating patient-derived cells and engineered microenvironments to enhance model relevance and support personalized medicine.The review also discusses advanced fabrication techniques such as micro-extrusion and laser-assisted bioprinting,which enable the creation of sophisticated tissue models capable of simulating critical GI processes,including molecular transport,peristalsis,and liver coupling.Advancing the complexity of in-vitro systems will help replicate the GI tract’s interactions and physiological phenomena,thus improving the translational potential of GI research.This review provides valuable insights into the advancements and challenges in GI modeling,serving as a comprehensive guide for developing models that bridge the gap between basic cell cultures and clinically relevant systems.展开更多
BACKGROUND Cardiovascular diseases are the first cause of death in the world.Ischemic heart disease is the main cause of heart failure.New approaches are continuously sou-ght to identify better therapeutic success.The...BACKGROUND Cardiovascular diseases are the first cause of death in the world.Ischemic heart disease is the main cause of heart failure.New approaches are continuously sou-ght to identify better therapeutic success.Thereby,current research has been drawn to identifying and completing the therapeutic profile of natural sources.Galium species are representatives exhibiting diuretic and antibacterial potential in living organisms and can treat burns,wounds,and skin diseases.Moreover,it was also observed that these plants manifest cardioprotective effects as well as having antihemolytic,antioxidant,antibacterial,anti-inflammatory,immunomod-ulatory,and antiproliferative potential.In ischemic heart disease,Galium verum(G.verum)extract manifested preservative properties in terms of contractility,systolic and diastolic function maintenance,and reduced damage to the heart after ischemia.In addition,G.verum extract upregulated the activity of antioxidant enzymes alleviating the production of pro-oxidants.AIM To test the ethanolic extract of G.verum on the H9C2(2-1)cell line by evaluating the in vitro biosafety profile and in ovo irritative potential.METHODS Cells were tested in vitro for viability(using the MTT test),cellular morphology,cell number,confluence,nuclear morphology(by immunofluorescence staining of cell nuclei and F-actin assay)and in ovo by the hen’s egg chorioal-lantoic membrane(CAM)test and CAM anti-irritant methods to study the irritation potential on the CAM.RESULTS The extract demonstrated a dose-dependent stimulatory activity.The viability increased to 170%for the dose of 55µg/mL and decreased to 135%at 200µg/mL.The results of cell number,confluence,and morphological analysis did not present significant changes compared with control untreated cells.The immunofluorescence assay showed insignificant apoptotic potential,and the hen’s egg CAM test revealed that the extract was in the weak to mode-rately irritating category with an irritation score of 5.3.When applying the sample to the CAM,only slight coagu-lation was observed(128 s).The anti-irritant test revealed the protective potential of the extract in the vascular plexus.CONCLUSION The ethanolic extract of G.verum manifests a stimulating effect on cardiomyocytes,enhancing cell viability,and maintaining a normal elongated shape,cell number,and confluence,without significant signs of apoptosis and with a weak irritative effect in ovo.In addition,the extract demonstrated a protective effect against hemorrhage,lysis,and coagulation of blood vessels induced by sodium dodecyl sulfate on the CAM.展开更多
Macrophages derived from the human THP-1 cell line have been widely used as substitutes for primary macrophages in various macrophage-related studies.However,difficulties still exist in establishing THP-I macrophage m...Macrophages derived from the human THP-1 cell line have been widely used as substitutes for primary macrophages in various macrophage-related studies.However,difficulties still exist in establishing THP-I macrophage models.This research presents techniques for generating different phenotypes of activated macrophages derived from THP-1 cells by introducing specific stimuli and provides some potential markers to confirm each type of activated macrophage.It is hoped to provide novel and useful methods for scientific research and to help researchers explore this field more intuitively and effectively.展开更多
文摘Alzheimer's disease (AD) is a progressive neurodegenerative disorder and is the most prominent cause of dementia.In 2019,over 57.4million people were living with AD and other dementia subtypes,a number which is expected to increase to over 152.8 million in the next 25years.This ever-increasing burden has resulted in AD and other neurodegenerative diseases rising to one of the top 10 causes of death globally (O'Connell et al.,2024).
基金part of the EU consortium DCog Plast ‘Diet Cognition and Plasticity” funded by the Joint Programming Initiative “A Health Diet for a Healthy Life”(JPI-HDHL) via the BMWFW (BMWFW-10.420/0009-WF/V/3c/2015 and the Medical Research Council UK:MR/N030087/1)(to LA and ST)supported by the PMU-FFF Research Fund (A-16/01/019-AIG)+9 种基金BA by the PMU-Research and Innovation Fund (PMU-RIF)(project 2023-PRE-008-Altendorfer)supported by the Center for Urban Mental Healthby Alzheimer Nederlandthe Zon MW Program Mechanisms Of DEMentia (MODEM)by the Gravitation program iCNS of the Dutch Research Council (NWO)supported by Grant PID2020-114921RB-C21Maria de Maeztu Unit of Excellence grant CEX2021-001234-M funded by MCIU/AEI/and CIBERFESCB16/10/00269, from the Instituto de Salud Carlos III all of them by “ERDF A way of making Europe”the Generalitat de Catalunya’s Agency AGAUR of 2021SGR00687ICREA Award
文摘In Alzheimer’s disease,microglial phagocytosis is engaged in the pathogenesis as it clears abnormal protein accumulations,debris,and apoptotic cells in the early stages of Alzheimer’s disease,but fuels neuroinflammation and accelerates disease progression in later stages.In vivo parabiosis experiments in aged animals have demonstrated that blood-born factors modulate synaptic plasticity,neurogenesis,and microglial responses.We hypothesize that peripheral factors can modulate microglial function and thereby possibly influence Alzheimer’s disease pathology.The objective of this study is to investigate the effects of Alzheimer’s disease serum on microglial phagocytosis.Here,we use an immortalized human microglial cell line in an in vitro parabiosis assay to investigate the impact of the serum from individuals diagnosed with Alzheimer’s disease(n=30)and age-matched controls(n=30)(PRODEM study)on microglial phagocytosis.Exposure to Alzheimer’s disease serum increased microglial phagocytic uptake of pH-sensitive fluorescent particles and downregulated expression of the lysosomal master regulator transcription factor EB(TFEB)and of ATPase H^(+)transporting lysosomal V1 subunit B2(ATP6V1B2),a component of the vacuolar ATPase.To identify serum components that may relate to changes in phagocytosis,serum samples of the Three-City Study(3C Study)were used.In the 3C Study,blood samples were collected up to 12 years before the onset of cognitive decline or dementia and their serum metabolome is well-defined.Microglia exposed to the serum of future Alzheimer’s disease patients from the 3C Study displayed an increased phagocytic uptake compared with the serum of matched controls,depending on the presence of the apolipoprotein Eε4 allele in the Alzheimer’s disease patients.Furthermore,microglial phagocytosis correlated inversely with serum levels of the omega-3 fatty acid eicosapentaenoic acid.We confirmed this inverse correlation between eicosapentaenoic acid and phagocytosis in the serum samples of the PRODEM cohort.In addition,in vitro testing of eicosapentaenoic acid on microglial phagocytosis showed a concentration-dependent decrease in phagocytic uptake.In conclusion,following incubation with Alzheimer’s disease blood serum,we observed increased microglial phagocytic uptake and the downregulation of TFEB and ATP6V1B2,possibly indicating lysosomal dysfunction.Furthermore,microglial phagocytosis was inversely correlated with serum eicosapentaenoic acid levels,suggesting an important role for dietary eicosapentaenoic acid in microglial function.
文摘This study aimed to optimization of the in vitro fertilization system in Cỏ goat oocytes to achieve the maximum possible blastocyst development rate. In Experiment 1, we assessed the effects of IVF media on the in vitro fertilization of Cỏ goat oocytes. There was no significant difference in the cleavage, blastocyst, or hatching rates between TALP-Fert and BO-IVF media. Experiment 2 was performed to assess the concentration of sperm in the in vitro fertilization of Cỏ goat oocytes. The matured Cỏ goat oocytes were fertilized in BO-IVF for four sperm concentrations: 5 × 105, 1 × 106, 2 × 106 and 3 × 106 sperm/ml. The blastocyst rate of 2 × 106 sperm/ml and 3 × 106 sperm/ml groups was higher than that of 5 × 105 sperm/ml and 1 × 106 sperm/ml groups (P Experiment 3 was performed to assess the IVF duration on the in vitro fertilization of Cỏ goat oocytes. The matured Cỏ goat oocytes were fertilized in BO-IVF with sperm concentration of 3 × 106 sperm/ml for 18, 20, 22 and 24 h. The cleavage, blastocyst, and hatching blastocyst rates of 18 h group were lower than those of 20, 22 and 24 h groups (P 0.05). In conclusion, the matured Cỏ goat oocytes were fertilized in BO-IVF with sperm concentration of 3 × 106 sperm/ml for 20 hours, which is suitable for the in vitro Cỏ goat embryo production.
文摘Objective:This study aims to investigate the energy conduction mechanism of holmium laser lithotripsy based on the Gemini platform and validate the optimal lithotripsy parameters under this mechanism using an in vitro ureteral model.Methods:The Lumenis VersaPulse PowerSuite dual-wavelength laser system was employed,with a holmium laser wavelength of 2.1μm,power output of 0-80 W,pulse energy ranging from 0.2 to 3.5 J,and pulse frequency from 5 to 40 Hz.An in vitro thermal effect model of the holmium laser was established to simulate temperature changes in urinary stones under varying power levels,energy ranges,pulse frequencies,and irrigation rates.High-speed cameras and thermal imaging devices were used to observe and record the temperature of the stone and surrounding tissues during laser irradiation.Finite element analysis was integrated to simulate the energy conduction pathways of the holmium laser within the stone and ureteral wall,assessing whether localized high-temperature regions exceeded the thermal injury threshold.Results:Temperature in the stone fragmentation mode decreased significantly with increasing distance from the basket and higher irrigation rates,thereby reducing the risk of thermal injury.In fragmentation mode,both 276μm and 500μm core-diameter fibers generated significantly higher temperatures at the basket site compared to the free-running mode(P<0.05).No thermal injury was observed at irrigation rates≥15 ml/min across all parameter combinations;at 10 ml/min,only high-energy settings(3.5 J)approached the thermal injury threshold(50.3℃);at≤5 ml/min,high-energy settings(>1.5 J)consistently exceeded the danger threshold.The safest parameter combination was 0.5 J+5 Hz(at any irrigation rate).Within 5 min,temperatures at the stone surface-below,middle,and above the basket-reached 151.0℃±1.0℃,62.5℃±0.2℃,and 46.8℃±0.2℃,respectively,whereas the maximum temperature on the ureteral wall surface was only 49.5℃.The FEA results closely matched experimental observations,indicating that with 0.5 J+5 Hz(at any flow rate),laser energy is primarily confined within the stone,generating localized high-temperature zones(80℃-90℃)at the stone-ureter wall interface,which promotes stone fragmentation while effectively minimizing collateral tissue damage.Conclusion:The holmium laser energy conduction mechanism based on the Gemini platform is predominantly concentrated within the stone,exhibiting excellent directionality and depth penetration,enabling efficient lithotripsy while minimizing thermal damage to surrounding tissues.
基金supported by grants from the National Key Research and Development Program of China(No.2018YFC1004201)National Natural Science Foundation of China(No.82473642)+3 种基金Research Funds of the Center for Big Data and Population Health of IHM(No.JKS2023017)Natural Science Foundation of Anhui Province(No.2408085QH278)Research Fund of the Anhui Institute of Translational Medicine(No.2022zhyx-C05)Anhui Medical University National Undergraduate Innovation and Entrepreneurship Training Program(No.S202410366070).
文摘Infertility,defined as the inability to achieve a clinical pregnancy after 1 year of unprotected and regular sexual intercourse,affects approximately 8%–12%of couples worldwide during their childbearing years.Assisted reproductive technology(ART)offers interventions to facilitate conception in couples with infertility.Since the inception of traditional in vitro fertilization(IVF)in 1978,ART has facilitated the birth of millions of infants.Intracytoplasmic sperm injection(ICSI)is an effective technique for couples ineligible for traditional IVF.Individual female factors,such as age and endometrial thickness,have been shown to affect embryo development during the IVF process,resulting in unfavorable pregnancy outcomes[1-3].However,the influence of environmental and male factors on early reproductive outcomes cannot be ignored.As a novel form of environmental pollution,light at night(LAN)has intensified with the rapid pace of urbanization,potentially leading to reproductive health problems in both women and men.However,to our knowledge,no study has explored the effects of LAN exposure on the early reproductive outcomes of IVF.Furthermore,the normal development of embryos relies on the contributions of both partners,and the influence of male factors on the early reproductive outcomes of IVF should not be overlooked.
文摘Correction to:Neurosci.Bull.December,2016,32(6):577–584.https://doi.org/10.1007/s12264-016-0068-z In this article,in Fig 5A,the picture of the Vector+Nogo-66 group was incorrect and should have appeared as shown below.
基金Funded by the National Key R&D Program of China(No.2023YFC2412300)the Guangdong Basic and Applied Basic Research Foundation(No.2021B1515120091)the Technology Development Project of Shandong Weigao Orthopedic Materials Co.,Ltd.(No.20221h0074)。
文摘This study employed a microwave-ultrasonic-hydrothermal multifield coupling method to synthesize nanoβ-Tricalcium phosphate(β-TCP)powder,systematically evaluating the impact of various parameters,including reaction temperature,time,sintering temperature,reactant types and concentrations,and graphene oxide(GO)concentration,on the physicochemical properties of the nanoβ-TCP powder.The synthesized powder was characterized using X-ray diffraction(XRD),Fourier-transform infrared spectroscopy(FTIR),scanning electron microscopy(SEM),high-resolution transmission electron microscopy(HRTEM),inductively coupled plasma optical emission spectroscopy(ICP-OES),and thermogravimetric-differential scanning calorimetry(TG-DSC).The experimental results indicate that the optimal synthesis conditions are achieved with a 0.6 mol/L Ca(NO3)2·4H2O solution and a 0.4 mol/L(NH_(4))_(2)HPO_(4)solution at a reaction temperature of 35℃for 40 minutes,followed by sintering at 720℃for 2 hours with 1×10^(-1)g/L GO.The preparedβ-TCP powder exhibits high crystallinity,a pure phase,good dispersibility,no significant aggregation,and uniform particle size of 59.75±12.84 nm.In vitro cytotoxicity tests show excellent biocompatibility and no cytotoxic effects on bone marrow stromal cells(BMSCs)even at concentrations up to 0.8 mg/mL.Furthermore,results from live-dead staining and nuclear membrane staining of cells co-cultured with the material demonstrate that theβ-TCP can promote the proliferation and differentiation of BMSCs to a certain extent,highlighting its potential as a safe and effective material for bone tissue engineering.
基金supported by the Volkswagen Foundation(Grant No.Az 99078 to DDC,ALT,and MT)funded by the Deutsche Forschungsgemeinschaft(DFG,German Research Foundation)under Germany’s Excellence Strategy–2082/1–390761711(to DDC)part of the research training group GRK 2415–Mechanobiology in Epithelial 3D Tissue Constructs(project number 363055819,to ALT and SJ).
文摘The global demand for in vitro respiratory airway models has surged due to the coronavirus disease 2019(COVID-19)pandemic.Current state-of-the-art models use polymer membranes to separate epithelial cells from other cell types,creating a nonphysiological barrier.In this study,we applied three-dimensional(3D)printing and bioprinting to develop an in vitro model where endothelial and epithelial cells were in direct contact,mimicking their natural arrangement.This proof-ofconcept model includes a culture chamber,with an endothelial bioink printed and perfused through an epithelial channel.In silico simulations of the air velocity within the channel revealed shear stress values ranging from 0.13 to 0.39 Pa,aligning with the desired in vivo shear stress observed in the bronchi regions(0.1–0.4 Pa).Biomechanical movements during resting breathing were mimicked by incorporating a textile mesh positioned away from the cell–cell interface.The epithelial channel demonstrated a capacity for compression and expansion of up to−14.7%and+6.4%,respectively.Microscopic images showed that the epithelial cells formed a uniform monolayer within the lumen of the channel close to the bioprinted endothelial cells.Our novel model offers a valuable tool for future research into respiratory diseases and potential treatments under conditions closely mimicking those in the lung.
文摘Objective:This review aims to investigate and establish potential in vitro and in vivo models for evaluating the anti-urolithiatic activity of therapeutic agents,exploring experimental approaches that can reliably simulate human stone formation and effectively assess the efficacy of anti-urolithiatic compounds.Methods:Multiple in vitro and in vivo approaches were explored.In vitro methods included the estimation of calcium oxalate by titrimetry,nucleation assays,aggregation assays,turbidimetric assays,and electron microscopy studies.Artificial stone models such as BegoStone and Ultracal 30 were fabricated to mimic the physicochemical characteristics of urinary calculi.In vivo models included ethylene glycol-induced,calcium oxalate/ammonium oxalate-induced,diet-induced,and infection-related models in rodents.Additionally,genetically modified animal models such as TRPV5 knockout,CLDN14 knockout,AGXT knockout,and URAT1 overexpression mice were discussed to study molecular pathways of urolithiasis.Parameters such as urinary oxalate,calcium levels,and histopathological evaluation of kidney tissues were used to validate stone formation and dissolution processes.Results:In vitro models effectively demonstrate the processes of crystal nucleation,aggregation,and inhibition,allowing quantitative assessment of potential anti-urolithiatic activity.Electron microscopy provides detailed insights into crystal morphology and ultrastructural alterations.Artificial stones fabricated using BegoStone and Ultracal 30 closely replicate natural calculi hardness and composition,making them suitable for lithotripsy and dissolution studies.In vivo models successfully mimic human urolithiasis pathophysiology,particularly the ethylene glycol-induced rat model,which shows reproducible calcium oxalate crystal deposition in renal tissues.The application of genetic models highlights the role of specific transporters and enzymes in calcium and oxalate homeostasis.Conclusion:A combination of in vitro and in vivo experimental models provides a comprehensive platform for evaluating the anti-urolithiatic potential of therapeutic agents.The integration of biochemical,morphological,and genetic analyses enhances the understanding of stone pathogenesis and development of novel anti-urolithiatic therapies.
文摘As prepubertal boys do not yet produce spermatozoa,they cannot rely on sperm cryopreservation for fertility preservation before gonadotoxic therapy,such as high-dose alkylating agents or radiotherapy in the case of childhood cancers.According to the current guidelines,cryopreservation of testicular biopsies containing spermatogonial stem cells(SSCs)may be proposed to high-risk patients for potential later therapeutic use to fulfill the patients’wish for a biological child.One promising technique for human in vitro spermatogenesis and in vitro propagation of human SSCs is microfluidic(MF)culture,in which cells or tissues are subjected to a continuous flow of medium.This provides exact control over such parameters as nutrient content and gradients,as well as the removal of waste metabolites.While MF has been shown to maintain tissues and cell populations of organs for longer than conventional in vitro culture techniques,it has not been widely used for testicular in vitro culture.MF could advance human testicular in vitro culture and is also applicable to reprotoxicity studies.This review summarizes the findings and achievements of testis-on-chip(ToC)setups to date and discusses the benefits and limitations of these for spermatogenesis in vitro and toxicity assessment.
基金supported by the National Key Research and Development Program of China(Grant Nos.2023YFA0915200 and 2023YFA0915204)Outstanding Young Scientist in Shandong Province(Grant No.ZR2024YQ064)+4 种基金Shandong Province Science and Technology Small and Medium Enterprises Innovation Capacity Improvement Project(Grant No.2024TSGC008)Shandong Provincial Key Research and Development Project(Grant Nos.2020CXGC011304 and 2022CXGC020206)Taishan Scholar Foundation of Shandong Province(Grant No.tsqn202408256)the Major Innovation Project for the Science Education Industry Integration Pilot Project of Qilu University of Technology(Shandong Academy of Sciences)(Grant No.2023JBZ03)Shanghai Science and Technology Development Funds(Grant No.23J21900100).
文摘Rapid and sensitive detection of targeted biomarkers in trace samples is of great significance for early in vitro diagnosis of diseases.Microfluidic technology has competitive advantages in this field due to its low cost,high efficiency,and high portability;however,the analysis of results tends to rely on bulky and sophisticated instruments,and this limits its applications.In this work,we developed a Raspberry Pi camera-based biomarker detection device based on microfluidic technology and digital image colorimetry.For highly sensitive biomarker detection on microfluidic chips,we propose a three-step signal-amplification colorimetric detection strategy consisting of:(1)the release of Ag^(+)ions from silver nanoparticles,(2)Ag^(+)-inhibited urea hydrolysis colorimetry,and(3)microscopic lens magnification.For efficient evaluation of results,we employed an RGB image-processing system to quantitatively analyze color images captured by the Raspberry Pi camera.Further,we tested the functionality of the device with procalcitonin(PCT)in phosphate-buffered saline,plasma,and serum to simulate clinical situations.We determined the limit of detection as 1 ng/ml,and a good linear relationship was established between PCT concentration and color intensity within the detection range 1–10 ng/ml.Importantly,only a relatively short detection time(40 min)was required in all three environments.The results demonstrate the great potential of this device for biomarker detection and facilitating biomedical research.
文摘The olive oil fermentation filtrate was prepared by microbial fermentation technology,and the fermentation process was optimized by orthogonal test to screen out the optimal fermentation conditions.At the same time,the effect of olive oil fermentation filtrate to promote the secretion of type I collagen (COL-I) and type VII collagen (COL-VII) by fibroblasts (HFF-1),the inhibitory effect on the secretion of alpha tumor necrosis factor (TNF-α) and interleukin-6 (IL-6) by lipopolysaccharide induced macrophages (RAW264.7),and the inhibitory effect on the induction of reactive oxygen species (ROS) production by immortalized human keratinocytes (HaCaT) were studied through in vitro cell testing methods.The results showed that the optimal fermentation conditions of olive oil fermentation filtrate were as follows,inoculation amount was 3%,inducer amount was 2%,fermentation time was 48 h.Its efficacy can significantly promote the synthesis of COL-I and COL-VII,and significantly inhibit the content of inflammatory factors TNF-α and IL-6 as well as ROS content,it has multiple effects of firming,anti-wrinkle,soothing and antioxidant,providing scientific and effective data support for the development of cosmetic raw materials.
基金supported by the National Natural Science Foundation of China(No.22306111)Shandong Provincial Natural Science Foundation of China(Nos.ZR2022QB242 and ZR2020ZD20).
文摘Considerable levels of methylmercury(MeHg)have been found rice-based infant cereals as a result of MeHg transfer from the rice as a raw material to the products,hence consumption of rice products may pose a potential health risk to infants who may receive cereals as the major diets and are susceptible to toxicity of MeHg.Determination of bioaccessibility of MeHg would provide a more accurate assessment of MeHg exposure through consumption of rice-based cereals,yet this information remains lacking.Further,the re-adsorption of methylmercury on the residual food will affect the accurate assessment of its bioaccessibility.Our goals in this work were 1)to determine the bioaccessibility of MeHg in infant rice cereals commonly available on the market by using a typical in vitro artificial gastrointestinal digestionmodel and 2)to evaluate the effects of MeHg re-adsorption on the in vitro assessment of MeHg bioaccessibility.The determined bioaccessibility of MeHg in the studied rice cereals after the standard dual-step(using gastric followed by intestinal juice)protocol ranged from 25%to 74%,a wide range comparable to that of fish samples observed here and in previous studies.The surprisingly higher bioaccessibility of MeHg in fish and rice cereals after the gastric step only,in comparison to after the complete two-step gastrointestinal digestion,suggests the re-adsorption of MeHg on the residual food.Separate experiments with spiked MeHg standards confirmed that the MeHg released by the acetic gastric juice was re-adsorbed on the residues during the intestinal step at neutral pH.This study provided first-hand data on the bioaccessibility of MeHg in infant rice cereals and methodological implications on using in vitro digestion to evaluate the bioaccessibility of MeHg and metal contaminants in general.
基金support of this research by the Program for Science&Technology Innovation Platform and Talent of Hunan Province(2021RC40322019TP1029)+2 种基金the Key R&D Plan of Hunan Province(2022SK2100,2023NK2034)the Ministry of Agriculture of the People’s Republic of China(GJFP20230204)the Forestry Science and Technology Innovation Project in Hunan Province(XLK202431).
文摘Edible mushroom polysaccharides demonstrate a range of bioactivities and have been investigated as potential adjuvant therapies for cancer treatment.Our prior research found Lactariks hatsudake Tanaka polysaccharide(LHP)effective against hepatocellular carcinoma,but the mechanisms remain unclear.This study aims to clarify the anti-cancer molecular mechanisms by examining the effects on specific signaling pathways through in vivo and in vitro analyses.Animal studies showed that LHP significantly inhibited the growth of transplanted liver cancer without toxicity.Histological and immunohistochemical analyses revealed that LHP increased nuclear cohesion in tumor cells and altered the expression of key regulatory proteins such as protein kinase B(Akt),nuclear factorκB(NFκB),caspase-3,p21,and p53.These findings suggest that LHP’s anticancer effects involve promoting apoptosis and disrupting the cell cycle.At the cellular level,assays such as 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS),flow cytometry,and transwell invasion confirmed that LHP inhibited cancer cell proliferation and induced apoptosis.Polymerase chain reaction and Western blot assays revealed that LHP intervention reduced cyclin-dependent kinase 4 activity by upregulating p53 and p21,leading to cell cycle arrest and inhibited cell proliferation.LHP also promoted apoptosis by altering Bcl-2-associated X protein,B-cell lymphoma 2,and cleaved caspase-3 levels.Moreover,LHP could affect the Akt,NF-κB,and mitogen-activated protein kinase signaling pathways to exert anticancer effects.These findings underscore the potential of LHP as a novel and multifaceted therapeutic agent.Furthermore,the elucidation of LHP-related mechanisms offers a crucial theoretical foundation for developing innovative and effective treatments for hepatocellular carcinoma.Additionally,this supports its potential development as a functional food or adjuvant therapy for cancer,which holds significant implications and application prospects in both the food and medical fields.
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Pimpinella anisum,commonly known as anise,is generally used in both folk medicine and the culinary world.In traditional medicine,it is valued for its digestive,respiratory,and antispasmodic properties.This study aims to examine the volatile compounds and antibacterial effect of P.anisum essential oil(PAEO)as well as for the first time its genotoxicity employing both in vitro and computational approaches.Gas chromatography-mass spectrometry(GC-MS)analysis identified anethole as the principal compound,which comprises 92.47%of PAEO.PAEO was tested for its potential antibacterial properties against Bacillus subtilis ATCC 6633,Listeria innocua ATCC 33090,Staphylococcus aureus ATCC 29213,Klebsiella aerogenes ATCC 13048,and a clinical strain of Salmonella enterica serotype Typhi.PAEO displayed noteworthy antibacterial action toward all tested bacteria,especially Staphylococcus aureus,with an inhibition zone of 21.43±0.87 mm,as determined by the disc-diffusion test.Varied between 0.0625%and 2%v/v,while the MBC values ranged from 0.125%to 8%v/v,reflecting the strength of the tested EO.The MBC/MIC ratios indicated the bactericidal nature of PAEO.The results of molecular docking revealed strong binding interactions between key PAEO molecules and microbial target proteins.ADMET(Absorption,Distribution,Metabolism,Excretion,and Toxicity)analysis confirmed favorable pharmacokinetic properties,indicating its potential as a safe therapeutic agent.Additionally,genotoxicity was assessed using the comet assay,which demonstrated minimal genotoxic risk,affirming the oil’s safety.These results highlight the promising antimicrobial properties of PAEO and its possible use as an active agent in the pharmacy,food,and cosmetic sectors.
文摘Phalaenopsis orchids are economically important ornamental crops;however,their commercial micropropagation is often limited by poor rooting efficiency and inconsistent growth.In this study,we investigated the effects of silver nanoparticles(Ag-NPs)on the in vitro regeneration and growth of Phalaenopsis cultivar 611B to determine the optimal concentration of Ag-NPs for improved micropropagation outcomes.Shoot tip explants(2–3 mm)—derived from protocorm-like bodies were cultured on a regeneration medium containing Hyponex(20:20:20 and 6.5:6.5:19),18 g/L sugar,2 g/L peptone,0.8 g/L activated charcoal,12.5 g/L potato extract,50 mL/L apple juice,and 10 mg/L 6-benzylaminopurine(6-BA),with varying concentrations of Ag-NPs(0,0.5,1.0,2.0,and 2.5 mg/L).After 10–12 weeks,shoot and root formation,plant height,fresh weight,leaf number,and chlorophyll contents were evaluated.At 1.0 mg/L Ag-NPs,shoot regeneration(5.4 vs.2.9 shoots per explant),root induction(2.1 vs.1.4 roots per explant),and shoot formation frequency(100%vs.55%)were significantly higher than the control(0 mg/L).Fresh weight(592.4 mg)and leaf number(9.7)also showed notable increases at this concentration.Although chlorophyll a and b levels peaked at 2.0 mg/L,the difference from 1.0 mg/L was not statistically significant.These results suggest that 1.0mg/LAg-NPs is the optimal concentration for enhancing shoot and root development and improving overall plantlet quality in Phalaenopsis.The findings highlight the potential of nanomaterials to improve the efficiency of orchid tissue culture systems.
基金Supported by Javna Agencija za Raziskovalno in Inovacijsko Dejavnost RS,No.P3-0036,No.I0-0029,No.J3-50098,and No.N1-0305University medical center Maribor,No.IRP-2021/02-05Swiss National Science Foundation(SNF)Postdoctoral Fellowship,No.P500PM217647/1.
文摘The gastrointestinal(GI)tract is essential for digestion,absorption,excretion,and protection,supported by a diverse microbial ecosystem.Traditional in-vitro models often fall short in capturing the physiological complexity of the GI tract,limiting their translational applications.A comprehensive approach is needed to bridge the gap between simple cell cultures and more complex systems used in translational research.This review explores the limitations of conventional twodimensional cell cultures and emphasizes the emerging use of three-dimensional and microfluidic systems that better replicate the GI tract’s structure and functions.It highlights the importance of incorporating patient-derived cells and engineered microenvironments to enhance model relevance and support personalized medicine.The review also discusses advanced fabrication techniques such as micro-extrusion and laser-assisted bioprinting,which enable the creation of sophisticated tissue models capable of simulating critical GI processes,including molecular transport,peristalsis,and liver coupling.Advancing the complexity of in-vitro systems will help replicate the GI tract’s interactions and physiological phenomena,thus improving the translational potential of GI research.This review provides valuable insights into the advancements and challenges in GI modeling,serving as a comprehensive guide for developing models that bridge the gap between basic cell cultures and clinically relevant systems.
文摘BACKGROUND Cardiovascular diseases are the first cause of death in the world.Ischemic heart disease is the main cause of heart failure.New approaches are continuously sou-ght to identify better therapeutic success.Thereby,current research has been drawn to identifying and completing the therapeutic profile of natural sources.Galium species are representatives exhibiting diuretic and antibacterial potential in living organisms and can treat burns,wounds,and skin diseases.Moreover,it was also observed that these plants manifest cardioprotective effects as well as having antihemolytic,antioxidant,antibacterial,anti-inflammatory,immunomod-ulatory,and antiproliferative potential.In ischemic heart disease,Galium verum(G.verum)extract manifested preservative properties in terms of contractility,systolic and diastolic function maintenance,and reduced damage to the heart after ischemia.In addition,G.verum extract upregulated the activity of antioxidant enzymes alleviating the production of pro-oxidants.AIM To test the ethanolic extract of G.verum on the H9C2(2-1)cell line by evaluating the in vitro biosafety profile and in ovo irritative potential.METHODS Cells were tested in vitro for viability(using the MTT test),cellular morphology,cell number,confluence,nuclear morphology(by immunofluorescence staining of cell nuclei and F-actin assay)and in ovo by the hen’s egg chorioal-lantoic membrane(CAM)test and CAM anti-irritant methods to study the irritation potential on the CAM.RESULTS The extract demonstrated a dose-dependent stimulatory activity.The viability increased to 170%for the dose of 55µg/mL and decreased to 135%at 200µg/mL.The results of cell number,confluence,and morphological analysis did not present significant changes compared with control untreated cells.The immunofluorescence assay showed insignificant apoptotic potential,and the hen’s egg CAM test revealed that the extract was in the weak to mode-rately irritating category with an irritation score of 5.3.When applying the sample to the CAM,only slight coagu-lation was observed(128 s).The anti-irritant test revealed the protective potential of the extract in the vascular plexus.CONCLUSION The ethanolic extract of G.verum manifests a stimulating effect on cardiomyocytes,enhancing cell viability,and maintaining a normal elongated shape,cell number,and confluence,without significant signs of apoptosis and with a weak irritative effect in ovo.In addition,the extract demonstrated a protective effect against hemorrhage,lysis,and coagulation of blood vessels induced by sodium dodecyl sulfate on the CAM.
基金supported by the Shaanxi University of Chinese Medicine-National Natural Science Foundation Cultivation Project(No.2023GP16)the Shaanxi Key Laboratory of Research on TCM Physical Constitution and Diseases Prevention and Treatment(No.KF202311)the Graduate Student Innovative Practical Ability Improvement Project(No.CXSJ202302),China.
文摘Macrophages derived from the human THP-1 cell line have been widely used as substitutes for primary macrophages in various macrophage-related studies.However,difficulties still exist in establishing THP-I macrophage models.This research presents techniques for generating different phenotypes of activated macrophages derived from THP-1 cells by introducing specific stimuli and provides some potential markers to confirm each type of activated macrophage.It is hoped to provide novel and useful methods for scientific research and to help researchers explore this field more intuitively and effectively.
