Isolation and short term cultivation of swine hepatocytes for bioartificial liver support system

BACKGROUND: The demand for the clinical use of hepa- tocytes is increasing. The aim of this study was to develop a method for procurement of high qualitative pig hepatocytes and to evaluate the state of freshly isolated and cultured hepatocytes. METHODS: The domestic extracorporeal circulating perfu- sion apparatus was used to isolate and harvest swine hepato- cytes by the two-step perfusion method with EDTA and collagenase. The viability, function and morphology of the freshly isolated and cultured cells were evaluated and ob- served by the trypan blue exclusion test, biochemical mea- surements, phase contrast microscopy and transmission electron micrography (TEM). RESULTS: The total yield of isolated hepatocytes reached to 1.5(±0.4)×l010 per liver with a viability of 92(±5)%, and the purity of hepatocytes reached to 98% Immediately after isolation, phase-contrast microscope and TEM showed that undamaged hepatocytes appeared bright, translucent and spherical in shape, with a characteristic well-contrasted border. After 24 hours, the concentrations of alanine aminotransferase (ALT), aspartate aminotrans- ferase ( AST ), lactate dehydrogenase ( LDH ), albumin (ALB), creatinine (Cr) and blood urea nitrogen (BUN) in the fluid of culture were declined significantly. CONCLUSION: This method of procuring swine hepato- cytes could get high quality cells with active metabolic function.

Mediating role of health literacy in the relationship between social isolation and psychological distress among pre-frail older adults

BACKGROUND Frailty has become a significant public health issue.The recent increase in the number of frail older adults has led to increased attention being paid to psycho-logical care services in communities.The social isolation of pre-frail older adults can impact their psychological distress.AIM To explore the mediating effect of health literacy between social isolation and psychological distress among communitydwelling older adults with pre-frailty.METHODS This descriptive cross-sectional study was conducted with 254 pre-frail older individuals aged 60 years and over.Social isolation,health literacy,and psycholo-gical distress were comprehensively measured using the Lubben Social Network Scale-6,12-item Short-Form Health Literacy Questionnaire,and the Kessler Psy-chological Distress Scale-10.Data were evaluated using the SPSS 27.0 package program and the PROCESS macro tool.Descriptive statistical analyses,correlation analyses,and bootstrap mediation tests were used to assess associations between the variables.RESULTS The results showed that social isolation had an effect on health literacy among pre-frail older adults(β=0.240,P<0.001),social isolation impact on psycho-logical distress pre-frail older adults(β=-0.415,P<0.001);health literacy was identified effect on psychological distress among pre-frail older persons(β=-0.307,P<0.001).Health literacy partially mediated the relationship between social isolation and psychological distress among community-dwelling older adults with pre-frailty,with a mediation effect of-0.074,accounting for 17.83%of the total effect.

Seismic Response Analysis of Steel Structure Isolation System Under Long-Period Seismic Motion

To analyze the seismic response of steel structure isolation systems under long-period seismic motion,a 9-story steel frame building was selected as the subject.Five steel structure finite element models were established using SAP2000.Response spectrum analysis was conducted on the seismic motion to determine if it adhered to the characteristics of long-period seismic motion.Modal analysis of each structural model revealed that the isolation structure significantly prolonged the structural natural vibration period and enhanced seismic performance.Base reactions and floor displacements of various structures notably increased under long-period seismic motion compared to regular seismic activity.Placing isolation bearings in the lower part of the structure proved more effective under long-period seismic motion.In seismic design engineering,it is essential to consider the impact of long-period seismic motion on structures and the potential failure of isolation bearings.

The response of roots and the rhizosphere environment to integrative cultivation practices in paddy rice

Integrative cultivation practices(ICPs)are essential for enhancing cereal yield and resource use efficiency.However,the effects of ICP on the rhizosphere environment and roots of paddy rice are still poorly understood.In this study,four rice varieties were produced in the field.Each variety was treated with six different cultivation techniques,including zero nitrogen application(0 N),local farmers’practice(LFP),nitrogen reduction(NR),and three progressive ICP techniques comprised of enhanced fertilizer N practice and increased plant density(ICP1),a treatment similar to ICP1 but with alternate wetting and moderate drying instead of continuous flooding(ICP2),and the same practices as ICP2 with the application of organic fertilizer(ICP3).The ICPs had greater grain production and nitrogen use efficiency than the other three methods.Root length,dry weight,root diameter,activity of root oxidation,root bleeding rate,zeatin and zeatin riboside compositions,and total organic acids in root exudates were elevated with the introduction of the successive cultivation practices.ICPs enhanced nitrate nitrogen,the activities of urease and invertase,and the diversity of microbes(bacteria)in rhizosphere and non-rhizosphere soil,while reducing the ammonium nitrogen content.The nutrient contents(ammonium nitrogen,total nitrogen,total potassium,total phosphorus,nitrate,and available phosphorus)and urease activity in rhizosphere soil were reduced in all treatments in comparison with the non-rhizosphere soil,but the invertase activity and bacterial diversity were greater.The main root morphology and physiology,and the ammonium nitrogen contents in rhizosphere soil at the primary stages were closely correlated with grain yield and internal nitrogen use efficiency.These findings suggest that the coordinated enhancement of the root system and the environment of the rhizosphere under integrative cultivation approaches may lead to higher rice production.

Plasma nitrogen fixation system with dual-loop enhancement for improved energy efficiency and its efficacy for lettuce cultivation

Plasma nitrogen fixation(PNF)has been emerging as a promising technology for greenhouse gasfree and renewable energy-based agriculture.Yet,most PNF studies seldom address practical application-specific issues.In this work,we present the development of a compact and automatic PNF system for on-site agricultural applications.The system utilized a gliding-arc discharge as the plasma source and employed a dual-loop design to generate NO_(x)from air and water under atmospheric conditions.Experimental results showed that the system with a dualloop design performs well in terms of energy costs and production rates.Optimal operational parameters for the system were determined through experimentation,resulting in an energy cost of 13.9 MJ mol^(-1)and an energy efficiency of 16 g kWh^(-1)for NO_(3)^(-)production,respectively.Moreover,the concentration of exhausted NO_(x)was below the emission standards.Soilless lettuce cultivation experiments demonstrated that NO_(x)^(-)produced by the PNF system could serve as liquid nitrate nitrogen fertilizer.Overall,our work demonstrates the potential of the developed PNF system for on-site application in the production of green-leaf vegetables.

Female zebra finches are more vulnerable to social isolation stress than males:Involvement of the nonapeptide system

In group-living animals,chronic juvenile social isolation stress(SIS)can profoundly affect behavior and neuroendocrine regulation.However,its impact on social behavior in avian species,particularly regarding sexspecific neural circuit differences,remains underexplored.This study focused on zebra finches,a species known for its social clustering and cognitive abilities,to elucidate these influences.Results indicated that SIS significantly increased plasma corticosterone levels in females but not in males,suggesting a heightened stress response and susceptibility in females.Additionally,SIS disrupted sociality and flocking behavior in both sexes,with more severe impairments in social recognition observed in females.Mesotocin(MT)levels in the lateral septum of both sexes and in the ventromedial hypothalamus of females were found to mediate the SIS effect,while vasotocin(VT)levels within the social behavior network remained unchanged.Pharmacological interventions confirmed the critical role of MT in reversing SIS-induced impairments in sociality,flocking behavior,and social recognition,particularly in females.These findings highlight unique nucleus-and sex-dependent variations in MT and VT regulation,providing novel insights into the mechanisms governing avian social behavior.This study advances our understanding of the independent evolutionary pathways of neural circuits and neuroendocrine systems that modulate social behaviors across different taxonomic groups.

Effects of water and nitrogen rate on grain-filling characteristics under high-low seedbed cultivation in winter wheat

A high-efficiency mode of high-low seedbed cultivation(HLSC)has been listed as the main agricultural technology to increase land utilization ratio and grain yield in Shandong Province,China.However,limited information is available on the optimized water and nitrogen management for yield formation,especially the grain-filling process,under HLSC mode.A three-year field experiment with four nitrogen rates and three irrigation rates of HLSC was conducted to reveal the response of grain-filling parameters,grain weight percentage of spike weight(GPS),spike moisture content(SMC),and winter wheat yield to water and nitrogen rates.The four nitrogen rates were N1(360 kg ha^(-1) pure N),N2(300 kg ha^(-1) pure N),N3(240 kg ha^(-1) pure N),and N4(180 kg ha^(-1) pure N),respectively,and the three irrigation quotas were W1(120 mm),W2(90 mm),and W3(60 mm),respectively.Results showed that the determinate growth function generally performed well in simulating the temporal dynamics of grain weight(0.989<R^(2)<0.999,where R2 is the determination coefficient).The occurrence time of maximum filling rate(T_(max))and active grain-filling period(AGP)increased with the increase in the water or nitrogen rate,whereas the average grain-filling rate(G_(mean))had a decreasing trend.The final 1,000-grain weight(FTGW)increased and then decreased with the increase in the nitrogen rates and increased with the increase in the irrigation rates.The GPS and SMC had a highly significant quadratic polynomial relationship with grain weight and days after anthesis.Nitrogen,irrigation,and year significantly affected the T_(max),AGP,G_(mean),and FTGW.Particularly,the AGP and FTGW were insignificantly different between high seedbed(HLSC-H)and low seedbed(HLSC-L)across the water and nitrogen levels.Moreover,the moderate water and nitrogen supply was more beneficial for grain yield,as well as for spike number and grain number per hectare.The principal component analysis indicated that combining 240-300 kg N ha^(-1) and 90^(-1)20 mm irrigation quota could improve grain-filling efficiency and yield for the HLSC-cultivated winter wheat.

Strawberry (Fragaria spp.): Cultivation, Production, Consumption, and Marketing in Cameroon

Strawberry (Fragaria spp.) is one of the most important fruits classified as exotic fruits imported into Cameroon. To have an inventory of its cultivation in Cameroon, a survey study was carried out among eight farms of Fragaria spp. from January 2021 to February 2022. The plant was introduced in Cameroon in 2018. There are 13 varieties of Fragaria spp. currently cultivated. Among these 13 varieties, eleven are hybrids of Fragaria x ananassa (“Amiga”, “Amine”, “Camarosa”, “Chandler”, “Charlotte”, “Elsanta”, “Gariguette”, “Madame Moutot”, “Ostara”, “Ruby gem” and “San Andreas”), and two of the hybrids of Fragaria vesca (“Maestro” and “Mara des bois”). The cropping system, irrigation system, and type of fertilizers applied differ from one strawberry farm to another. Biofertilizers (such as mycorrhizal), inorganic and organic fertilizers are actually used to improve production. The potential annual production of strawberries from January 2021 to February 2022, estimated based on the survey data, was 21.216 tons for all growers. Among these eight production farms, the Lolodorf BIO Farm presents 6000 kg (six tons) of strawberries and 100,000 stolons (seedlings) produced, from seven varieties of Fragaria spp. cultivated, with 6 varieties which are hybrids variety Fragaria x ananassa (“Amiga”, “Amine”, “Chandler”, “Gariguette”, “Madame Moutot”, and “Ruby gem”), and one which is a hybrid of Fragaria vesca (“Mara des bois”). Certain diseases were also observed and recorded depending on the growing areas.

Cultivation Path of Nursing Undergraduates’Scientific Research and Innovation Ability

Objective: The cultivation of the innovation ability and scientific research is one of the nursing learning objectives for undergraduate students. To explore the method and effect of training system of scientific research innovation ability of nursing undergraduates based on “3332”. Methods: Three course learning modules are constructed: stage-based course learning module, systematic project practice training module and comprehensive practice training module. A practical training platform for scientific research innovation projects is built, and undergraduate scientific research innovation ability training is carried out from both in-class and out-of-class lines. Results: Since 2017, the students have obtained 7 national innovation and entrepreneurship training programs, 52 university-level undergraduate scientific research projects, published more than 10 academic papers, and obtained 2 patent authorization. Conclusions: The training system of scientific research innovation ability of nursing undergraduates based on “3332” is conducive to the development of scientific research innovation ability of nursing students, and to cultivate nursing talents who can adapt to the development of the new era and have better post competence.

Detection of Bisphenol A (BPA) in Plastic Bottles Using Vertical Cultivation at Various Temperatures

Polycarbonate plastics containing bisphenol A (BPA) used to manufacture drinking water bottles. Kurdistan region in northern Iraq is a developed area with increased pollution from plastic bottles. Trace amounts of BPA have been detected in bottled water samples. The absorption of BPA was measured with HPLC using a vertical cultivation system with Bulbs of the Allium Cepa plant planted in these plastic bottles with monitored growth. Vertical cultivation was found to have a low level of BPA in the plant cells, making it a safe cultivation method under specific climate conditions. The mean concentration of BPA in vertical cultivation is 0.19 ug/ml (3.8 ng for a 20 uL injection), and the Limit of Quantification (LOQ) is 0.63 ug/ml (12.7 ng for 20 uL injection). While Scanning Electron Microscope (SEM) shows that the concentrations are relatively low in water samples stored at room temperature compared to those exposed to direct sunlight (40°C) and water bottle samples stored at (-4°C), The correlation coefficients were found to be good (0.9992). SEM is used for plastic bottle samples stored at different temperatures. The images identify compound decay and explore the morphology of BPA in manufactured plastic materials.

Determinants of Tobacco Cultivation and the Associated Impacts of Adoption in Hurungwe District, Zimbabwe

Tobacco is an essential cash crop in Zimbabwe and a strategic livelihood option for hundreds of thousands of rural households. However, the crop is linked to negative environmental, economic, and social impacts. The existing studies on tobacco cultivation in Zimbabwe present contradictory findings on the determinants and impacts of adoption, leaving unanswered questions about the crop’s sustainability impact in the country. This article investigates the determinants of smallholder farmers’ decisions to grow tobacco and the associated impacts of adoption. Random and purposive sampling were used to select 273 household surveys, including tobacco and non-tobacco smallholder farmers, and 56 expert interviews to answer the research questions. We employed regression models alongside expert interviews and document analysis to identify the determinants influencing the decision-making process of smallholder farmers in Zimbabwe regarding tobacco cultivation. Additionally, our investigation aimed to elucidate the perceived impacts associated with the adoption of this agricultural practice. The regression analysis indicated that the farmer’s age, education level, farming experience, family size, household income, and perceived high farm profitability are significant drivers of tobacco adoption. We also discovered divergent and convergent perceptions of the critical impacts of tobacco cultivation. The study highlights the need for proactive multi-stakeholder collaboration and sustainable financial arrangements to address the negative impacts of tobacco production. As the primary stakeholder responsible for regulating and promoting agricultural activities, the Zimbabwean government should provide meaningful financial support, increase access to credit, and ensure better market facilities for alternative crops to reduce the over-dependence on tobacco.

Technique of Earthworms Restoring Soil in Greenhouse Cultivation

The production environment of greenhouse cultivation is relatively closed,the multiple cropping index is high,the management of fertilizationwatering and pesticideapplication isblindtosomeextent,andthe phenomenonofcontinuous cropping isalsocommonSoilquali-ty affects the sustainable development of greenhouse cultivation.Earthworm is a ubiquitous invertebrate organism in soil,an important part of soil system,a link between terrestrial organisms and soil organisms,an important link in the small cycle of soil material organisms,and plays an important role in maintaining the structure and function of soil ecosystem.Different ecotypes of earthworms are closely related to their habi-tats(soil layers)and food resource preferences,and then affect their ecological functions.The principle of earthworm regulating soil function is essentially the close connection and interaction between earthworm and soil microorganism.Using different ecotypes of earthworms and bio-logical agents to carry out combined remediation of greenhouse cultivation soil is a technical model to realize sustainable development of green-house cultivation.

Strategic insights into the cultivation of pancreatic cancer organoids from endoscopic ultrasonography-guided biopsy tissue

BACKGROUND The frequent suboptimal efficacy of endoscopic ultrasound-guided fine-needle biopsy(EUS-FNB)to culture pancreatic cancer(PC)organoids(PCOs)poses a major challenge in the advancement of personalized medicine for advanced PC.AIM To explore how to obtain appropriate puncture tissues from EUS-FNB and optimize the strategy for efficiently constructing PCOs,providing an efficient tool for the advancement of personalized medicine.METHODS Patients who underwent EUS-FNB for the diagnosis of PC tissue were prospectively enrolled.We refined the endoscopic biopsy procedures and organoid cultivation techniques.All tissue specimens verified by on-site pathological assessment were cultured in a semi-suspended medium in a microfluidic environment.We assessed differences in PCOs cultured beyond and below five generations examining patient demographics,specimen and organoid attributes,and the sensitivity of organoids to a panel of clinical drugs through cell viability assays.RESULTS In this study,16 patients with PC were recruited,one sample was excluded because onsite cytopathology showed no tumor cells.Successful organoid generation occurred in 93.3%(14 of 15)of the EUS-FNB specimens,with 60%(9 of 15)sustaining over five generations.Among these patients,those with a history of diabetes,familial cancer,or larger tumors exhibited enhanced PCO expandability.The key factors influencing longterm PCOs expansion included initial needle sample quality(P=0.005),rapid initiation of organoid culture postisolation(P≤0.001),and high organoid activity(P=0.031).Drug sensitivity analysis revealed a partial response in two patients following therapeutic intervention and surgery and stable disease in four patients,indicating a moderate correlation between organoid response and clinical outcomes.CONCLUSION Optimal initial needle sampling,rapid and precise biopsy sample processing,process isolated samples as soon as possible,and sufficient cellular material are crucial for successful cultivating PCOs.High organoid activity is an important factor in maintaining their long-term expansion,which is essential for shortening the time of drug sensitivity analysis and is the basis of PC research.

Identification,Isolation and Genomic Characterization of Porcine Astrovirus in Shandong Province,China in 2021-2023

[Objective]The paper was to identify,isolate,and characterize porcine astrovirus in Shandong Province between 2021 and 2023.[Method]A total of 1025 samples of porcine diarrhea samples were collected from various regions of Shandong Province between January 2021 and October 2023.The samples were tested by RT-PCR,followed by sequencing and phylogenetic analyses of the polymerase.[Result]The total positive rate of PAstV was 34.6%(355/1025).The respective proportions of individuals infected with PAstV-1,PAstV-2,PAstV-4 and PAstV-5 were 25.4%(90/355),28.2%(100/355),35.2%(125/355)and 22.5%(80/355),respectively.Additionally,mixed infection was observed.Meanwhile,849 samples of healthy pigs were tested by RT-PCR,and the results demonstrated that the total positive rate of PAstV was 8.13%(69/849).Of these,the proportion of PAstV-1,PAstV-2 and PAstV-4 infection was 27.5%(19/69),37.7%(26/69)and 40.6%(28/69),and a mixed infection also existed.Further sequencing and characterization of some the selected isolates revealed low sequence identities(56.2%)with known PAstV strains,indicating the presence of novel types or genotypes of PAstVs.Furthermore,the isolation conditions of porcine astrovirus were optimized,resulting in the purification of a pure PAstV-4 strain(designated PAstV-4-GRF1).The virus was found to exhibit typical astroviral morphology,with nucleotide identity ranging from 89.9 to 95.4%with previously published PAstV-4 strains.Then,macrovirus transcriptome sequencing showed that 88.30%of the CRF1 samples were mammalian astroviruses.By species classification,PAstV 4 and PAstV 2 accounted for 21.79%and 0.32%,respectively.Phylogenetic tree analysis showed that the c15050 fragment was identical to the GRF-1 sequencing fragment of the isolated strain,and exhibited the highest homology with the Hunan PAstV-4 sequence MK460231 in China.[Conclusion]As the inaugural isolated PAstV-4 strain,it furnishes pivotal materialfor the investigation ofthe biological and pathogenic properties of this virus as well as for the prospectivedevelopment of relevant biological and diagnostic reagents.

Near resonance vibration isolation on a levered-dual response(LEDAR)Coulomb-damped system by differential preloads/offsets in linear springs

The levered-dual response(LEDAR)Coulomb-damped system attains near resonant vibration isolation by differential preloads/offsets in linear springs.It takes the advantages of both the preloads/offsets in linear springs and the guiderail friction for realizing different levels of vibration isolation.The isolation capacities are investigated on the strategies with both the horizontal and vertical guiderails,with the horizontal rail only,and without guiderails.The compressive preloads generally result in the consumption of most of the initial excitation energy so as to overcome the potential threshold.The isolation onsets at the frequency ratio of 1∓0.095 on the left-hand side(LHS)and the right-hand side(RHS)of the lever are relative to the load plate connector.The observed near resonant isolation thus makes the LEDAR system a candidate for the isolation of the mechanical systems about resonance while opening a path for simultaneous harvesterisolation functions and passive functions at extreme frequencies.

High Yield Cultivation and Pest Control Technology of Cashew

In order to enhance the yield and quality of cashew,it is essential to implement high-yield cultivation techniques effectively throughout the production process.Additionally,pest control measures should be employed to provide technical support for the industrialized development of cashew.

Rice Cultivation under Film Mulching Can Improve Soil Environment and Be Beneficial for Rice Production in China

Rice cultivation under film mulching is an integrated management technology that can conserve water, increase soil temperature, improve yield, and enhance water and nitrogen use efficiencies. Despite these advantages, the system does have its drawbacks, such as soil organic matter reduction and microplastic pollution, which impede the widespread adoption of film mulching cultivation in China. Nonetheless, the advent of degradable film, controlled-release fertilizer, organic fertilizer, and film mulching machinery is promoting the development of rice film mulching cultivation. This review outlines the impact of rice cultivation under film mulching on soil moisture, soil temperature, soil fertility, greenhouse gas emissions, weed control, and disease and pest management. It also elucidates the mechanism of changes in rice growth, yield and quality, water use efficiency, and nitrogen use efficiency. This paper incorporates a review of published research articles and discusses some uncertainties and shortcomings associated with rice cultivation under film mulching. Consequently, prospective research directions for the technology of rice film mulching cultivation are outlined, and recommendations for future research into rice cultivation under film mulching are proposed.

Refining adipose-derived stem cell isolation for optimal regenerative therapy

This article highlights the importance of optimizing the techniques used for isolating stromal vascular fraction cells from adipose tissue.Furthermore,by presenting key findings from the literature,it clarifies the effects of refined techniques on regenerative medicine and advocates for ongoing research and innovation to enhance therapeutic outcomes.

Variety Selection and Green Cultivation Technology of Courtyard Ornamental and Edible Peppers

In order to comply with the development trend of the multifunctional use of peppers,we conducted an investigation into the characteristics and features of varieties,potting management techniques,and the methods of extending the fruit ornamental period and other aspects of courtyard ornamental and edible peppers.A set of cultivation techniques suitable for courtyard ornamental and edible peppers has been developed,including timely sowing and seedling,nutrient soil preparation,water and fertilizer management,trimming and pruning,preservation of flowers and fruits,green prevention and control of diseases and pests,harvesting,and so on.

Entrepreneurial Spirit for China in the New Era and Strategies for Cultivation

What type of entrepreneurial spirit is needed for China’s new era,and how can we unlock its innovation potential?Answers to these questions will unleash vital forces to drive China’s high-quality economic development.After reviewing the theories,dimensions,features,and limitations of the entrepreneurial spirit in the Western context,we concluded that the entrepreneurial spirit in China’s new era,which is characterized by patriotism,innovation,trustworthiness,adherence to the law,fulfillment of social responsibilities,and a global horizon,has overcome the limitations that come with self-interested entrepreneurship in the context of Western economics.This is a major step forward in the development of theories on entrepreneurial spirit.In the development context of the new era,this paper argues that the entrepreneurial spirit should become an important force at the individual level to implement the new development concept,foster the new development paradigm,and promote high-quality development in the new stage.This paper provides answers to the question of what type of entrepreneurial spirit China’s new era requires.Finally,this paper presents basic strategies for cultivating entrepreneurial spirit in the context of China’s new era,emphasizing the importance to promote the healthy growth of young entrepreneurs,ensure their business autonomy,develop a multi-tiered capital market,implement the factor market reforms,improve the business climate,and accelerate the digital transition.