Long-term in-vitro culture and subculture of the hemocytes of swimming crab Portunus trituberculatus

Crab cell line,especially continuous crab cell line,can provide us a useful tool for studies on the virology,immunology,and molecular biology of crabs.However,no continuous crab cell line has been available due to the lacking of suitable medium and the occurrence of mitosis-arrest.In this study,long-term in vitro culture conditions for both two-(2D)and three-dimensions(3D)were successfully developed for the circulating hemocytes of swimming crab Portunus trituberculatus,designated as PTH cells.In 2D culture,a novel crab basic medium in osmolarity of 990–1100 mOsm/kg was optimized for the first time,which is different from Leibovitz's L-15 medium in mainly the components of amino acids,containing double strengths of the contents of free amino acid mixture in the crab serum.Then an optimal crab growth medium was developed by supplementing 5%fetal bovine serum,50-g/L yeast extract powder,20-μg/L basic fibroblast growth factor and epidermal growth factor into the optimal crab basic medium,and found that it could support a long-term survival of PTH cells in a healthy monolayer up to 347 days and partially break through the mitosis-arrest of crab cells evidenced by the obvious increase of proliferating potential detected in the 10-d primarily cultured PTH cells.These 2D cultured PTH cells could be successfully sub-cultured for 11 times by physical flushing method and well cryopreserved in liquid nitrogen.In 3D culture,using the same crab growth medium,the PTH cell aggregates could be easily formed and healthily maintained on the surface of solidified Matrigel or in the ultra-low-attachment plate with a survival rate of 50%–60%on Day 103.This work largely improved the primary culture and subculture of crab cells and will facilitate the establishment of continuous crab cell line.

Effects of Four Chemicals on Tissue Differentiation of Lespedeza bicolor in In-vitro Culture

In order to find the optimal bacteriostatic agents and selectors and their concentrations in genetic transformation of Lespedeza bicolor with BADH as a marker gene, an resistance experiment was carried out with four chemicals, NaCl, betaine aldehyde, PEG and cefotaxime （Cef） added into the media, Different concentrations and combinations of different chemicals were added into the media in the stage of adventitious bud regeneration of cotyledonary nodes, Shoot segment multiplication, and rooting. The results showed that NaCI and betaine aldehyde could inhibit tissue differentiation of L. bicolor, and could thus be used as selectors. Furthermore, 0.8 g/L NaCI could completely inhibit the cotytedonary node differentiation and shoot segment multiplication, and 0.5 g/L NaCI was optimal for inhibition of rooting. It was also found that with the concentration increase of betaine aldehyde in media, adventitious buds regenerated from cotyledonary nodes decreased, and 1.5 g/L was confirmed to be the optimum concentration. Moreover, the mixture of 0.7 g/L NaCI and 0.3 g/L betaine aldehyde significantly inhibited shoot segment multiplication and rooting, so this mixture could be used asa selector. The Cef ex- periment showed that 200-300 mg/L C, ef in media did not inhibit significantly the regeneration of adventitious buds from cotyledonary nodes, and 100 mg/L Cef had little effect on the shoot segment multiplication and rooting.

Key Technology of In-vitro Culture for a New Vaccinium uliginosum Cultivar Zishuijing

Objective] This study aimed to discuss the key technology of in-vitro cul-ture for a new Vaccinium uliginosum cultivar Zishuijing. [Method] Through the screen-ing and optimization of sterilization method for explants, sampling time, multiplication, nursing and rooting culture, a matching clone propagation system was established for the new Vaccinium uliginosum cultivar Zishuijing. [Result] The explants were sterilized with 0.1% HgCl2 for 3 min; the differentiation and multiplication medium of Zishuijing was composed of WPM （modified）, 6-BA （1.0 mg/L） and ZT （1.0 MG/L）; the rootless tube seedlings were transplanted in organic matrix （sawdust∶bark∶peat=1∶1∶1） in September and cultured at air relative humidity of 80%-90% and temperature of 20-25 ℃, and after 50 d, the rooting rate reached 72.4%. [Conclusion] The key technol-ogy of in-vitro culture for the new Vaccinium uliginosum cultivar Zishuijing was estab-lished, thereby providing technical support for large-scale industrialized seedling pro-duction of Zishuijing.

Comparison on In-Vitro Culture Methods of Yak Endometrial Gland Epithelial Cells

[ Objective] To develop in-vitro culture methods of yak endometrial gland epithelial cells. [Method] The gland epithelial cells were isolated from yak endometrium by explant culture method and digestion culture method, respectively. [ Result] In the first method, the cells isolated from the endometrium explant could merge into monolayer after 8-d culture, and they could be purified by gradation digestion with trypsin. In the second method, the endometrium explant were first digested by collagenase II by incubation at 37℃ for 2.5 h and then further digested in fresh 2 g/L colla- genase II for another 2.5 h. The cell suspension was leached through 74 pm filter and centrifuged at 400 r/min for 5 min. Then the cell pellet was re-suspended, followed by natural sedimentation to collect purified gland epithelial cells. The isolated cells were cytokeratin-positive as detected by immunocytochemical staining, and the positive rate could reach 95%. [Conclusion] The yak endometrial gland epithelial cells can be isolated and purified by both the explant culture method and digestion culture method.

Advances in In-vitro Culture Technology of Human Fetal Hepatic Stellate Cells

Liver is an important organ for human metabolism and biological conversion. Medical research on hepatic disease clinic, drug metabolism and drug effi- cacy evaluation all needs an in-vitro model of liver as a research platform. Hepatic stellate cells are core cells for occurrence and development of liver fibrosis. Stud- ies at home and abroad deemed that human fetal hepatic stellate cells are an ideal material for the construction of an in-vitro research model for liver fibrosis. With clinical and basic research of liver going deeper, the requirements to quantity and quality of in-vitro models of fetal hepatic stellate cells become higher and higher. The advances in isolation, culture and cryopreservation technique of human fetal hepatic stellate cells were reviewed in this paper.

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.

Investigation of the Acceptability of Cultured Meat in University Students

Background: Over the past 20 years, cultured meat has drawn a lot of public attention as a potential solution to issues with animal husbandry, including inadequate use of natural sources, improper animal welfare practices, and possible risks to public health and safety. The novel method of producing meat through culture reduces the need for animals to produce muscle fiber, thereby obviating the necessity for animal slaughter. Apart from its ethical advantages, cultured meat presents a possible way to fulfill the expanding need for food among growing populations. The purpose of this research was to find out whether Turkish students would be willing to pay for and accept cultured meat. Technique: Method: 371 university students who willingly consented to fill out a questionnaire and provide demographic data make up the research sample. Questions from previous studies on the acceptability of cultured meat were compiled to create the survey. The research’s data collection took place in March and April of 2022. The research was completed in June 2022 after the data had been processed and analyzed. Results: The results showed that the majority of participants were female and had omnivorous eating habits. Based on the results of the Bonferroni correction test, students with a higher intention to purchase and consume cultured meat were those who received economics and business education. Students with two years of university education had a higher overall survey score than those with four years of education (p < 0.05). Furthermore, it is discovered that there is a negative correlation between the participants’ ages and their Factor 2 (using cultured meat as an alternative to industrial meat) and Factor 3 (consuming and purchasing it) section points (r = -109, p = 0.036) (r = -0.121, p = 0.019). In conclusion, university students generally have a negative outlook on health-related issues, such as eating cultured meat as an alternative.

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.

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.

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.

Comparative Study of the Widal Test against Stool Culture in the Diagnosis of Suspected Cases of Typhoid Fever in Some Low Income Communities in the Adamawa Region of Cameroon

Introduction: Infectious diseases constitute a major concern of public health in developing countries. Facilities and well trained staff have been shown to be one of the major obstacles in the rapid and quality diagnosis of these diseases. As such, we carried out an analysis to compare the Widal test and stool culture to identify febrile patients with Salmonella infection. Method: A cross sectional study was conducted to diagnose salmonella infection with out-patients who demonstrated signs of salmonella infection. Serum was harvested from blood collected from 368 (Vina = 234, Mayo Banyo 65, and Djerem = 69) patients accompanied by stool, Widal test was conducted on the spot and stool was taken to a reference laboratory for culture using standard microbiological methods, sociological set up was calculated in percentages, prevalence was calculated using excel while statistical difference was calculated using SPSS. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated to compare the Widal test against stool culture. Results: A total of 368 (50.8% females and 49.2% males) participants took part in the survey. Salmonella prevalence (66.24%) in stool culture in the Vina division was significantly different (p 0.05). The sensitivity,specificity, PPV, and NPV of slide agglutination test against stool culture varied from different areas (Vina: 51.6%, 73.62%, 79.21% and 43.61%;Mayo Banyo: 60.53%, 77.78%, 79.31% and 58.33%;Djerem: 53.18%, 83.73% 73.91% and 67.39%) respectively. Slide agglutination test has a fair agreement with the stool culture (kappa, Vina = 0.202;Mayo Banyo = 0.37 and Djerem = 0.38). Conclusion: Generally, in the three areas of study, the Widal test had a fair correlation with the stool culture;This means the Widal test should not be used alone but in combination with stool culture in the detection of salmonella infections.

Historiography and Scientific Research of Religious Buildings of the Cucuteni-Trypillian Culture

The purpose of this article is to reveal the historiography and current state of research related to the cult buildings of the Eneolithic period of the Cucuteni-Trypillian cultural community.The article describes the historical path of archaeological discoveries,from the first Trypillian sanctuaries to the discovery of the largest Nebelivka temple complex in Ukraine in 2012.At the same time,the work partially raises the issue of religious beliefs of the ancient farmers of Central Europe in connection with the discovery of their sacred buildings.Since special scientific works have not comprehensively addressed this issue and are still fragmentary in nature,it is important at the present stage of research to make some coverage and systematization of existing materials on this issue.

AIGC Related Context: A New Communication Culture For Human

The emergence and rapid development of Artificial Intelligence Generated Content(AIGC)technology is revolutionizing human communication and cultural landscapes.This paper explores the impact of AIGC on society,identifying three distinct cultural paradigms:AIGC Blended Culture,AIGC Based Culture,and AIGC Polluted Culture.AIGC Blended Culture represents a synergistic collaboration between AI and human creativity,enhancing content creation efficiency,promoting cross-cultural communication,and enriching human experiences.AIGC Based Culture signifies a shift where AI becomes the core driver of cultural production and dissemination.This leads to innovative cultural industry models,expanded development fields,and improved cultural product quality and diversity.However,AIGC Polluted Culture arises from the misuse of AIGC to produce or disseminate harmful content.The paper proposes countermeasures and recommendations to maximize the benefits of AIGC while mitigating potential risks.This involves fostering collaboration between AI and humans,focusing on talent cultivation,establishing and improving laws and regulations,and developing solutions to detect and mitigate harmful content.Additionally,promoting responsible innovation and ethical AI development is crucial.In conclusion,AIGC represents a transformative force reshaping human communication and culture.While offering immense opportunities for innovation and efficiency,it also presents challenges that require careful consideration and proactive measures to ensure its positive and sustainable impact on society.

Effect of Butternut Squash (Cucurbita moschata) Seed Powder on the Chemical and Rheological Properties of Stirred Cultured Camel Milk and Yoghurt

Research shows that producing fermented camel milk is hard because of the milk’s inability to form a firm coagulum, attributed to low levels of κ-casein and ꞵ-lactoglobulin and the large casein micelle size, leading to a weak network of casein formation. In an effort to address this issue, researchers turned to corn starch as a thickening agent, discovering that a concentration of 2.0% effectively improved the viscosity and significantly reduced syneresis in stirred camel milk yoghurt and cultured camel milk. This study explores alternatives to corn starch, focusing on butternut squash seeds as a promising substitute due to their hydrocolloid composition. By incorporating butternut squash (Cucurbita moschata) seed powder (BSSP) as a thickening agent, this study aimed at enhancing the chemical and rheological properties of stirred camel milk yoghurt and cultured camel milk. Fermented camel milk was prepared using 4 litres of camel milk, 2% starter cultures (thermophilic culture for yoghurt and mesophilic aromatic culture for stirred cultured camel milk) and BSSP 0.0% (negative control), 0.4%, 0.8%, 1.2%, 1.6%, 2.0% mixed with 0.4% gelatin. 2.0% corn starch mixed with 0.4% gelatin was used as a standard for comparison. Results showed that increasing the BSSP level significantly (p < 0.05) decreased the moisture content while increasing the total solid content of stirred fermented camel milk products. There was an increase in ash content with an increase in BSSP levels. There was a significant (p < 0.05) reduction in the pH, with an increase in BSSP levels in stirred fermented camel milk samples. Increasing the concentration of BSSP from 0.4% to 2.0% resulted in a significant (p < 0.05) increase in viscosity and a reduction in syneresis of stirred camel milk yoghurt and stirred cultured camel milk samples. This study demonstrated that BSSP effectively enhances the viscosity, reduces syneresis and increases acidity in stirred fermented camel milk products during storage.

Tourism-Oriented Cultural Product Design: A Case Study of Yami Culture in Lanyu

Due to the prevalence of modern commerce and mainstream culture, many indigenous cultures around the world are rapidly diminishing. These cultures, with their unique primitiveness distinct from modern culture, often attract tourists. This study integrates the indigenous Yami culture of Lanyu Island, Taiwan Region, into innovative designs from the perspective of tourists’ understanding of the local culture. In this way, tourists can experience the harmonious coexistence of the Yami tribe and nature, while these culturally rich products foster deeper experiential and emotional resonance. This study employs the Mandala thinking method for concept expansion and the Osborn checklist method for the analysis and transformation of design elements. Subsequently, designers were invited to conduct on-site travel experiences, leading to the creation of a series of Yami cultural products. Through the evaluation and validation of cultural product assessment indicators, functional commercialized products were ultimately produced. This study aims to establish a design process for cultural products through this design case. It seeks to help users recall their Lanyu Island travel experiences and appreciate the often-overlooked cultural beauty, thereby raising awareness of cultural preservation.

Integrating Chinese Culture Into Language Curriculum:Teaching Chinese Culture to International Students in China

This paper explores the integration of Chinese culture into language education for German students at the University of Shanghai for Science and Technology(USST).Focusing on USST’s Chinese curriculum and pedagogical strategies,the study emphasizes the importance of cultural immersion,experiential learning,and authentic materials.Drawing on Byram’s Intercultural Communicative Competence(ICC)model,the Cultural Studies Approach,and Task-Based Language Teaching(TBLT),the paper presents a case study on incorporating Chinese calligraphy into regular classes.This hands-on approach not only enriches cultural understanding but also enhances language skills.The findings stress the need for tailored,multifaceted pedagogical approaches to prepare international students for cross-cultural interactions in a globalized context.

The effect of"rural cultural auditorium"on the revitalization of rural culture:A case study of villages A and B in Zhejiang Province

The construction of rural cultural auditorium is conducive to gathering local popularity,excavating history and continuing civilization,and it has become a powerful driving force for cultural revitalization in rural revitalization.However,the development of"Rural Cultural Auditorium"encountered many problems so far.From the perspective of rural cultural revitalization,and under the background of the new socialist rural cultural construction in China,this paper takes the rural cultural auditorium as the topic.The field questionnaire survey and semi-structured interview were constructed in Village A and Village B of Zhejiang Province,.SPSS software is used to analyze the collected questionnaire data.On the basis of combing related concepts and theories,it summs up the problems of"rural cultural auditorium",such as the existence of cultural auditorium,low participation rate of cultural auditorium activities,poor information communication,etc.Through visiting and investigating,going deep into the local area,according solutions are put forward such as increasing content diversity,paying attention to publicity and increasing interaction,so as to promote the healthy and sustainable development of rural cultural auditorium.It is hoped that this paper will explore the significance of the construction of rural cultural auditorium to the revitalization of rural culture in China,and put forward ideas on how to further build rural cultural auditorium,so as to provide reference for rural cultural construction services.

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.