Three-dimensional models:from cell culture to Patient-Derived Organoid and its application to future liposarcoma research

Liposarcoma is one of the most common soft tissue sarcomas,however,its occurrence rate is still rare compared to other cancers.Due to its rarity,in vitro experiments are an essential approach to elucidate liposarcoma pathobiology.Conventional cell culture-based research(2D cell culture)is still playing a pivotal role,while several shortcomings have been recently under discussion.In vivo,mouse models are usually adopted for pre-clinical analyses with expectations to overcome the issues of 2D cell culture.However,they do not fully recapitulate human dedifferentiated liposarcoma(DDLPS)characteristics.Therefore,three-dimensional(3D)culture systems have been the recent research focus in the cell biology field with the expectation to overcome at the same time the disadvantages of 2D cell culture and in vivo animal models and fill in the gap between them.Given the liposarcoma rarity,we believe that 3D cell culture techniques,including 3D cell cultures/co-cultures,and Patient-Derived tumor Organoids(PDOs),represent a promising approach to facilitate liposarcoma investigation and elucidate its molecular mechanisms and effective therapy development.In this review,we first provide a general overview of 3D cell cultures compared to 2D cell cultures.We then focus on one of the recent 3D cell culture applications,Patient-Derived Organoids(PDOs),summarizing and discussing several PDO methodologies.Finally,we discuss the current and future applications of PDOs to sarcoma,particularly in the field of liposarcoma.

Extended protective effects of three dimensional cultured human mesenchymal stromal cells in a neuroinflammation model

BACKGROUND Human mesenchymal stromal cells(MSCs)possess regenerative potential due to pluripotency and paracrine functions.However,their stemness and immunomod-ulatory capabilities are sub-optimal in conventional two-dimensional(2D)culture.AIM To enhance the efficiency and therapeutic efficacy of MSCs,an in vivo-like 3D culture condition was applied.METHODS MSCs were cultured on polystyrene(2D)or in a gellan gum-based 3D system.In vitro,prostaglandin-endoperoxide synthase 2,indoleamine-2,3-dioxygenase,heme oxygenase 1,and prostaglandin E synthase gene expression was quantified by quantitative real-time polymerase chain reaction.MSCs were incubated with lipopolysaccharide(LPS)-treated mouse splenocytes,and prostaglandin E2 and tumor necrosis factor-alpha levels were measured by enzyme linked immuno-sorbent assay.In vivo,LPS was injected into the lateral ventricle of mouse brain,and MSCs were administered intravenously the next day.Animals were sacrificed and analyzed on days 2 and 6.RESULTS Gellan gum polymer-based 3D culture significantly increased expression of octamer-binding transcription factor 4 and Nanog homeobox stemness markers in human MSCs compared to 2D culture.This 3D environment also heightened expression of cyclooxygenase-2 and heme-oxygenase 1,enzymes known for immunomodulatory functions,including production of prostaglandins and heme degradation,respectively.MSCs in 3D culture secreted more prostaglandin E2 and effectively suppressed tumor necrosis factor-alpha release from LPS-stimulated splenocytes and surpassed the efficiency of MSCs cultured in 2D.In a murine neuroinflammation model,intravenous injection of 3D-cultured MSCs significantly reduced ionized calcium-binding adaptor molecule 1 and glial fibrillary acidic protein expression,mitigating chronic inflammation more effectively than 2D-cultured MSCs.CONCLUSION The microenvironment established in 3D culture serves as an in vivo mimetic,enhancing the immunomodulatory function of MSCs.This suggests that engineered MSCs hold significant promise a potent tool for cell therapy.

Effects of Chlorine-Based Fertilizers on Tomato Growth under Soilless Culture

This study investigated the effects of chlorine-based fertilizers under varying nitrogen solution concentrations in a soilless culture system. The experiment included four nitrogen solution concentration levels, with nitrogen concentrations of 6 mmol/L (C1), 12 mmol/L (C2), 18 mmol/L (C3), and 24 mmol/L (C4). Each nutrient concentration level was further divided into four chloride ion treatments (R1, R2, R3, and R4), where 100%, 60%, 33%, and 0% of the NH4+ and K+ ions were derived from NH4Cl and KCl, respectively. The length, surface area and volume of root were significantly higher by 25.3%~136.9%, 40.1%~173.1%, 27.9%~178.0%, respectively, in the R4 treatment than in the R1 and R2 treatments at flowering stage. The aboveground biomass and yield in the R4 treatment were significantly higher, by approximately 15.6%~43.5% and 16.6%~28.6%, respectively, than in the R1, R2, and R3 treatments at the picking stage. The C3 and C4 treatments significantly decreased biomass and yield by 31.9%~50.2% and 20.7%~50.5%, respectively, compared to the C1 and C2 treatments at the picking stage. Besides, the higher nutrient solution increased the incidence of blossom-end rot. In conclusion, high concentrations of chloride ions in nutrient solutions, especially when the Cl− concentration exceeded 10 mmol/L, have been shown to inhibit tomato growth in soilless culture systems. Therefore, replacing sulfur-based fertilizers by chlorine-based fertilizers is not recommended for tomato production under the studied conditions.

“Culture time”板块教学中跨文化意识培养的策略

在新课程改革的大环境中,文化意识成为英语学科核心素养的重要组成部分。语言是文化的载体,语言与文化是相互依赖、相互影响、相互制约的。学习语言的过程也是学习文化的过程。文章主要结合译林版英语五、六年级教材中的Culture time板块的教学案例,探讨如何在英语课堂教学实践中丰富文化素材、发掘其中的文化元素和领略中西方文化异同,以帮助学生有效学习文化知识,拓宽文化视野,培养文化意识。

Preparation of lactic acid bacteria compound starter cultures based on pasting properties and its improvement of glutinous rice flour and dough

The effects of 5 lactic acid bacteria(LAB)fermentation on the pasting properties of glutinous rice flour were compared,and suitable fermentation strains were selected based on the changes of viscosity,setback value,and breakdown value to prepare LAB compound starter cultures.The results revealed that Latilactobacillus sakei HSD004 and Lacticaseibacillus rhamnosus HSD005 had apparent advantages in increasing the viscosity and reducing the setback and breakdown values of glutinous rice flour.In particular,the compound starter created using the two abovementioned LAB in the ratio of 3:1 had better performance than that using a single LAB in improving the pasting properties and increasing the water and oil absorption capacity of glutinous rice flour.Moreover,the gelatinization enthalpy of the fermented samples increased significantly.For frozen glutinous rice dough stored for 28 days,the viscoelasticity of frozen dough prepared by compound starter was better than that of control dough,and the freezable water content was lower than that of control dough.These results indicate that compound LAB fermentation is a promising technology in the glutinous rice-based food processing industry,which has significance for its application.

Anticancer Activity of Rice Callus Suspension Cultures from Aromatic Varieties and Metabolites Regulated in Treated Cancer Cell Lines

Tissue culture techniques were used to produce large amounts of bioactive compounds with medicinal potential, overcoming space and time constraints for cancer prevention. Rice callus suspension cultures(RCSC) and seed extracts prepared from aromatic rice varieties were used to evaluate the cytotoxic impact on human colon and lung cancer cell lines, as well as a normal control cell line, using Taxol as a positive control. RCSC and seed extracts from two Indian aromatic rice varieties were applied at different concentrations to treat the cancer cell lines and normal lung fibroblasts over varying time intervals. Apoptosis was assessed in 1:5 dilutions of the A549 and HT-29 cell lines treated with RCSC for 72 h, using propidium iodide staining and flow cytometry. RCSC showed a more potent cytotoxic effect than seed extracts with minimal effect on the normal cell line, in contrast to Taxol. Confocal microscopy and flow cytometry further confirmed the apoptotic effect of RCSC. Gas chromatography-mass spectrometry-based metabolic profiling identified metabolites involved in cytotoxicity and highlighted altered pathways. RCSC is proposed as an alternative source for the development of novel anticancer drugs with reduced side effects.

Effects of Different Culture Conditions and Growth States on the Structure and Hemostatic Properties of Coscinodiscus sp.

As a renewable marine inorganic material,Coscinodiscus sp.has significant potential in the field of rapid hemostasis.However,the low yield of Coscinodiscus sp.seriously limits the application.In this study,two new culture modes were adopted to increase the production of Coscinodiscus sp.,the effect of changes in culture conditions and growth status on the hemostatic activity of diatoms was detected.To prevent Coscinodiscus sp.from sinking in culture,the suspension culture mode was realized by adding0.5%agar.The semi-continuous high nutrient concentration culture mode increased the cell density of Coscinodiscus sp.to 11000cells mL^(-1)and shorten the culture cycle to 5 d.In terms of coagulation activity,the addition of frustules reduced the in vitro coagulation time by half and the activation time of coagulation by 70%.The hemolysis rate and cytotoxicity of frustules harvested in the two culture modes did not change significantly.The results showed that suspension culture mode and high nutrient concentration culture mode only changed the growth state of Coscinodiscus sp.,while the hemostatic performance remained stable.

Current status and challenges for cell-cultured milk technology: a systematic review

Cellular agriculture is an innovative technology for manufacturing sustainable agricultural products as an alternative to traditional agriculture.While most cellular agriculture is predominantly centered on the production of cultured meat,there is a growing demand for an understanding of the production techniques involved in dairy products within cellular agriculture.This review focuses on the current status of cellular agriculture in the dairy sector and technical challenges for cell-cultured milk production.Cellular agriculture technology in the dairy sector has been classified into fermentation-based and animal cell culture-based cellular agriculture.Currently,various companies synthesize milk components through precision fermentation technology.Nevertheless,several startup companies are pursuing animal cell-based technology,driven by public concerns regarding genetically modified organisms in precision fermentation technology.Hence,this review offers an up-to-date exploration of animal cell-based cellular agriculture to produce milk components,specifically emphasizing the structural,functional,and productive aspects of mammary epithelial cells,providing new information for industry and academia.

Efficacy of serum-free cultured human umbilical cord mesenchymal stem cells in the treatment of knee osteoarthritis in mice

BACKGROUND We investigated the efficacy of intra-articular injection of human umbilical cord mesenchymal stem cells(hUC-MSCs)for the treatment of osteoarthritis(OA)progression in the knee joint.Although many experimental studies of hUC-MSCs have been published,these studies have mainly used fetal bovine serumcontaining cultures of hUC-MSCs;serum-free cultures generally avoid the shortcomings of serum-containing cultures and are not subject to ethical limitations,have a wide range of prospects for clinical application,and provide a basis or animal experimentation for clinical experiments.AIM To study the therapeutic effects of serum-free hUC-MSCs(N-hUCMSCs)in a mouse model of knee OA.METHODS Fifty-five male C57BL/6 mice were randomly divided into six groups:The blank control group,model control group,serum-containing hUC-MSCs(S-hUCMSC)group,N-hUCMSC group and hyaluronic acid(HA)group.After 9 weeks of modeling,the serum levels of interleukin(IL)-1β and IL-1 were determined.Hematoxylin-eosin staining was used to observe the cartilage tissue,and the Mankin score was determined.Immunohistochemistry and western blotting were used to determine the expression of collagen type II,matrix metalloproteinase(MMP)-1 and MMP-13.RESULTS The Mankin score and serum IL-1 and IL-1β and cartilage tissue MMP-1 and MMP-13 expression were significantly greater in the experimental group than in the blank control group(P<0.05).Collagen II expression in the experimental group was significantly lower than that in the blank control group(P<0.05).The Mankin score and serum IL-1 and IL-1β and cartilage tissue MMP-1 and MMP-13 levels the experimental group were lower than those in the model control group(P<0.05).Collagen II expression in the experimental group was significantly greater than that in the model control group(P<0.05).CONCLUSION N-hUCMSC treatment significantly alleviate the pathological damage caused by OA.The treatment effects of the ShUCMSC group and HA group were similar.

The Philosophy of He(和):Examined from the Perspectives of Technology and Culture

Pre-modern Chinese crafts,such as iron smelting,cookery,medicine,and the production of vehicles,bows,and arrows indicate the traditional Chinese view of technology as being organic,holistic,and comprehensive.This view of technology is guided by the concept of he(和)and employs the means and methods of he,thus achieving the purport of he.In Chinese,the character he(和)possesses positive connotations.It originated from the meaning of"to season;to add flavoring to"(调和)and that of flavors being"perfectly harmonious"(和美).From this sensory experience,he gradually extended to the abstract levels of materiality,humanity,sociality,"order"(wei位),and "power,situation,force"(shi势).Finally,he was elevated to the supreme level of"qi of great harmony"(taihe zhi qi太和之气),which is comparable to the concept of dao(道).The philosophy of he has exerted wide impact on such areas as technology,art,national character,cultural psychology,and behavior patterns,and has become an integral part of China's inherent culture.The paradoxes and deviations of he hold their own profound philosophical implications that merit further exploration.As humanity confronts significant challenges,such as how we can coexist with others,with technology,and with nature,the ancient Eastern wisdom embodied in he continues to possess practical characteristics and value.

The Implantation and Transmission of Plague Culture by the Five Plague Gods in Shuilu Murals

The Five Plague Gods(五瘟神) in the Shuilu(水陆) murals from the Ming and Qing dynasties exemplify a distinctive manifestation of plague gods image developed under the pedigree of Taoist gods.As a significant system for expelling disasters in Shuilu,the depiction of the Five Plague Gods prominently merges human beings and livestock.This shape form to a large extent,draws on the image of Nuo(傩),a figure reflecting an imagination of the host of plague pathogens.In terms of the numbers of gods,it aligns with the conceptual framework of Wu Xing(五行 the five elements),a foundational principle in traditional Chinese medicine.The internal cultural connection highlights the means of plague prevention and control in China,as well as the external reflection of plague culture.Through multi-dimensional interpretations with the double proof method,this article dissects the formation,core elements,and cultural characteristics of the beliefs in the Five Plague Gods reflected in the Shuilu murals,and transcends mere visual semantics,offering a deeper understanding of these complex cultural symbols.

MACS-W:A modified optical clearing agent for imaging 3D cell cultures

Three-dimensional(3D)cell cultures have contributed to a variety of biological research fields by filling the gap between monolayers and animal models.The modern optical sectioning microscopic methods make it possible to probe the complexity of 3D cell cultures but are limited by the inherent opaqueness.While tissue optical clearing methods have emerged as powerful tools for investigating whole-mount tissues in 3D,they often have limitations,such as being too harsh for fragile 3D cell cultures,requiring complex handling protocols,or inducing tissue deformation with shrinkage or expansion.To address this issue,we proposed a modified optical clearing method for 3D cell cultures,called MACS-W,which is simple,highly efficient,and morphology-preserving.In our evaluation of MACS-W,we found that it exhibits excellent clearing capability in just 10 min,with minimal deformation,and helps drug evaluation on tumor spheroids.In summary,MACS-W is a fast,minimally-deformative and fluorescence compatible clearing method that has the potential to be widely used in the studies of 3D cell cultures.

Protective Effect of Dietary Yeast Cultures on Lipopolysaccharide-induced Oxidative Stress,Immune and Inflammatory Responses in Pseudobagrus ussuriensis

This study aimed to evaluate the effects of dietary yeast culture(YC)on lipopolysaccharide(LPS)-induced oxidative stress,immune and inflammatory response in P.ussuriensis.The fish were randomly assigned into three groups as the control group,LPS group and YC+LPS group.The fish in the control were fed diet with no YC supplementation and no LPS challenge,and the fish in the LPS group or YC+LPS group were fed diet supplemented with no YC or 20 g·kg^(-1)YC,and with LPS challenge,respectively.The results showed that compared with the control group,intestinal total antioxidant capacity(T-AOC)level and superoxide dismutase(SOD)activity were significantly decreased,while intestinal malondialdehyde(MDA),plasma aspartate transaminase(AST)and alanine transaminase(ALT)levels were significantly increased in the LPS group(P<0.05).Besides,lower plasma alkaline phosphatase(ALP),alternative complement pathway(ACH50)activity and the albumin(ALB)level,as well as higher lysozyme(LZM)activity,were also found in the LPS group.However,dietary 20 g·kg^(-1)YC supplementation could relieve the above LPS-induced changes in Pseudobagrus ussuriensis.Furtherly,LPS challenge could significantly up-regulate gene expression of interleukin-8(IL-8),heat shock protein(HSP70)and NF-κBp65 except for toll-like receptors 2(TLR2),while dietary 20 g·kg^(-1)YC supplementation suppressed the increased expression of NF-κBp65 and IL-8 induced by LPS in P.ussuriensis.In summary,LPS challenge could induce immune impairment,oxidative stress and hepatic damage,and the protective effect of dietary 20 g·kg^(-1)YC supplementation on LPS-induced immune impairment and oxidative stress was observed in the present study,which was associated with the enhanced levels of antioxidant enzymes and immune parameters.Also,dietary 20 g·kg^(-1)YC supplementation could suppress LPS-induced inflammatory response by down-regulating NF-κBp65,HSP70 and IL-8 gene expression.

Metabolic Characteristics of Taste Differences of Sweet Potato Leaves Grown in Soil and Hydroponic Cultures by Using Non-Targeted Metabolomics

Sweet potato leaves are consumed globally for their nutritional and functional properties,with their taste characteristics significantly influencing their market value and consumer acceptance.However,the metabolic factors determining the taste of sweet potato leaves remain unclear.This study evaluated the taste and metabolic profile of sweet potato leaves cultivated in soil and hydroponic systems,revealing that hydroponic culture improved the taste of the leafy sweet potatoes.Using gas chromatography system(GC)-ToF-MS,200 metabolites were identified,encompassing most plant metabolic pathways.A comparison of good-tasting vs.poor-tasting sweet potato leaves identified 71 metabolites associated with taste quality formation.Poor-tasting leaves exhibited lower levels of amino acid metabolites and higher levels of carbohydrate and secondary metabolites.This research provides novel insights into enhancing the taste of leafy sweet potatoes.

Construction of the Demonstration Area for the Inheritance and Development of Yimeng Red Culture Based on Digital Empowerment

Yimeng red culture is an important part of the excellent culture of the Chinese nation.It is of great practical significance and long-term strategic significance to build the demonstration area for the inheritance and development of Yimeng red culture.Combining digital empowerment with the inheritance and development of Yimeng red culture is a new idea that conforms to the development of the times.In this paper,on the basis of combing Yimeng red cultural resources and combining with the current situation of inheritance and development of Yimeng red culture,the strategic layout and implementation approach of the demonstration area for the inheritance and development of Yimeng red culture were put forward from the perspective of digital empowerment.

Establishment of an in vitro Tissue Culture and Rapid Propagation System using Lonicera hypoglauca and Content Assessment of Total Flavonoids and Chlorogenic Acid

Lonicera hypoglauca is a traditional Chinese medicinal plant.In this study,the tender young leaves of L.hypoglauca were used for the first time as the explants to establish a rapid in vitro propagation and regeneration system.The results revealed that the optimal time for disinfection of the explants was 8 min and the optimal medium for callus induction was MS+2,4-D 4.0 mg·L^(-1)+sucrose 30 g·L^(-1),with an average callus induction rate of 86.67%.The optimal medium to induce differentiation of callus to bud was MS+6-BA 1.0 mg·L^(-1)+NAA 0.10 mg·L^(-1)+sucrose 30 g·L^(-1),with an average germination rate of 83.33%.The optimal medium to induce multiplication was MS+6-BA 1.5 mg·L^(-1)+NAA 0.05 mg·L^(-1)+sucrose 30 g·L^(-1),with a multiplication coefficient of 5.42.The optimal medium for root induction was 1/2 MS+NAA 0.15 mg·L^(-1)+activated carbon 0.3 g·L^(-1)+sucrose 15 g·L^(-1),with an average rooting rate of 91.11%.The survival rate of tissue-cultured seedlings in nutrient soil cultivation medium was as high as 100%.The total flavonoid content and chlorogenic acid content in the explant,callus tissue and regenerated plant were 1.83%,2.27%,1.33%and 2.77%,1.83%,1.74%respectively.This study provides novel insights into the rapid propagation and mass production of L.hypoglauca seedlings at an industrial scale and that it exhibits important application value and future prospects.

Coupling coordination analysis of Mazu culture,socio-economy and ecological environment of Meizhou Island

Taking the pilgrimage,tourism and cultural island of Meizhou Island as an example,the evaluation index system of the coupling and coordinated development of“Mazu culture,socio-economy,eco-environment”(MSE)compound system was constructed.The index weights were determined by AHP-entropy method,and the coupling degree,coordinated degree,comprehensive evaluation index and grey correlation degree of MSE system of Mazu Island from 2012 to 2022 were measure.The results showed that:(1)the comprehensive evaluation indexes of the three subsystems was on the rise in general,but the evaluation index of the ecological subsystems increased relatively slowly.(2)The coupling degree was only in the running-in stage in 2012,and the other years were in the coordinated coupling stage.(3)The coupling coordination degree increased from 0.35 in 2012 to 0.82 in 2022,the coupling coordination level was changed from mild imbalance to good coordination.(4)Through the comparison of grey correlation degree,the 24 indexes in the evaluation index system had great influence on the coupling coordination degree of MSE system.The coupling coordination degree was closely related to the development of socio-economy and the spread of Mazu culture.With the rapid development of tourism brought about by the spread of Mazu culture,the pressure on the ecological environment will be increasing.Compared with the rapid growth of tourism and economy,it is equally important to strengthen environmental protection and pay attention to the quality of ecological environment development.

Screening of Substrate Formulas for Culture of Auricularia auricula Mycelia with Byproducts

[Objectives]This study was conducted to explore the source of substitutive substrate raw materials for cultivation of edible fungi,reduce the cutting and use of woody materials,and realize harmless treatment and resource utilization of byproducts from Chinese medicine production and bamboo processing.[Methods]Traditional Chinese medicine residue and bamboo shavings were used to partially replace hardwood sawdust in the conventional formula,and the growth of Auricularia auricula mycelia in large test tubes under different substrate formulas was studied.[Results]The results showed that the mycelia of A.auricula could grow normally on substrates with byproducts,and the mycelia grew differently with different formulas,and the performance of different strains of the same species was also different.Compared with the conventional formula,the suitable substitution amount of bamboo shavings for A.auricula strains was 10%-30%,and the substitution amount of Chinese medicine residue was 5%-15%.[Conclusions]This study provides reference for the efficient utilization of byproducts and the expansion of raw material sources for production of edible fungi.

Happiness and harmony through health culture diplomacy as integrative maternal-child health nursing

For this editorial,our colleague Dr.Naeema Hasan Al Qasseer,former World Health Organization(WHO)Senior Scientist of Nursing and Midwifery joins me to add her wisdom and experience on the topic of integrative maternal-child health nursing.

Tourism Mode of Tea Culture Museum in Guizhou:A Case Study of Meitan Modern Tea Culture Museum Settlement

Since the 14 th Five-Year Plan,tea culture has become very popular.However,due to the lack of brand awareness and overall planning,and the mechanical application of the model outside the province and other tourism products,Guizhou Tea Culture Tourism presents a"strong tea,light culture,weak tourism".In view of this,in order to realize the positioning goal of"building a national demonstration zone for the integration of tea culture and tourism"in the 14 th Five-Year Plan of Guizhou tea industry,and explore the development mode of integration of tea culture and tourism with Guizhou characteristics,this study takes museum tourism as the breakthrough point,Meitan Tea Museum settlement as the research object,and collects Meitan Tea Museum tourism resources.This paper uses SWOT analysis to evaluate the Meitan Tea Culture Museum Tourism model,and provides some recommendations for the development of tea culture museum tourism in Guizhou Province.