Fluorescent Tracer Technique Research Articles

Subserosal injection of indocyanine green fluorescence tracing technique in laparoscopic lymph node dissection for gastric cancer: a preliminary study

Subserosal injection of indocyanine green fluorescence tracing technique in laparoscopic lymph node dissection for gastric cancer: a preliminary study

Therapeutic impacts of GNE‑477‑loaded H2O2 stimulus‑responsive dodecanoic acid‑phenylborate ester‑dextran polymeric micelles on osteosarcoma.

Osteosarcoma (OS) is a highly malignant primary bone neoplasm that is the leading cause of cancer‑associated death in young people. GNE‑477 belongs to the second generation of mTOR inhibitors and possesses promising potential in the treatment of OS but dose tolerance and drug toxicity limit its development and utilization. The present study aimed to prepare a novel H2O2 stimulus‑responsive dodecanoic acid (DA)‑phenylborate ester‑dextran (DA‑B‑DEX) polymeric micelle delivery system for GNE‑477 and evaluate its efficacy. The polymer micelles were characterized by morphology, size and critical micelle concentration. The GNE‑477 loaded DA‑B‑DEX (GNE‑477@DBD) tumor‑targeting drug delivery system was established and the release of GNE‑477 was measured. The cellular uptake of GNE‑477@DBD by three OS cell lines (MG‑63, U2OS and 143B cells) was analyzed utilizing a fluorescent tracer technique. The hydroxylated DA‑B was successfully grafted onto dextran at a grafting rate of 3%, suitable for forming amphiphilic micelles. Following exposure to H2O2, the DA‑B‑DEX micelles ruptured and released the drug rapidly, leading to increased uptake of GNE‑477@DBD by cells with sustained release of GNE‑477. The in vitro experiments, including MTT assay, flow cytometry, western blotting and RT‑qPCR, demonstrated that GNE‑477@DBD inhibited tumor cell viability, arrested cell cycle in G1 phase, induced apoptosis and blocked the PI3K/Akt/mTOR cascade response. In vivo, through the observation of mice tumor growth and the results of H&E staining, the GNE‑477@DBD group exhibited more positive therapeutic outcomes than the free drug group with almost no adverse effects on other organs. In conclusion, H2O2‑responsive DA‑B‑DEX presents a promising delivery system for hydrophobic anti‑tumor drugs for OS therapy.

Fluorescent Tracing of Dialdehyde Sodium Alginate Tanning Agent in Leather Matrix

Dialdehyde polysaccharides possess a sustainable nature and good tanning performance. However, the lack of specific detectable groups in their molecular structure results in the difficulty in the determination of the location of dialdehyde polysaccharides in leather fiber networks. In this study, dialdehyde sodium alginate (DSA) tanning agent was fluorescent labeled by 5-(4,6-dichlorotriazinyl) aminofluorescein (DTAF). The purified DTAF–DSA showed high and stable fluorescent intensity at emission wavelength of 515.2 nm when the pH was over 6.0, and the temperature was in the range of 20°C to 50°C. DTAF–DSA was used in tanning and tracked using fluorescence microscopy. Its penetration in the fiber network could be clearly visualized, and its distribution in leather differed with the molecular weight of DSA. As a result, this fluorescence tracing technique could display the mass transfer behavior of dialdehyde polysaccharide tanning agents in leather matrix, which will provide underlying data for tanning mechanism exploration.

Research progress in the evaluation of glymphatic system function by the DTI-ALPS method.

The glymphatic system can remove metabolic wastes from the brain, which plays a significant role in maintaining the homeostasis of the central nervous system. It is an important basis for advanced cognitive functions such as learning and memory. Studies have analyzed the function of glymphatic system by diffusion tensor imaging analysis along the perivascular space (DTI-ALPS) recently. Compared with other invasive examinations that require fluorescent tracer technique or the injection of contrast agents, DTI-ALPS can evaluate the hydromechanics of the glymphatic system via quantifying the diffusion rate of water molecules in different directions, which turns out to be a non-invasive in vivo neuroimaging method. The ALPS-index calculated by the DTI-ALPS method is significantly correlated with the cognitive function in diseases of central nervous system and other system and can reflect the dynamic changes of diseases. In general, ALPS-index is expected to become a novel neuroimaging biomarker for predicting prognosis and clinical effects.

Membrane anatomy-based radical resection for esophageal cancer

Radical resection of gastrointestinal tumors based on the membrane anatomy theory has significantly reduced the postoperative recurrence rate and improved the surgical efficacy. However, the theory of membrane anatomy has not been widely adopted in radical surgery for esophageal cancer. Our study found that the esophagus also has a membranous anatomical structure. As a foregut organ, the esophagus also has a mesenteric structure, and there is also a fifth metastasis pathway within the esophageal mesentery for esophageal cancers. The leak and metastasis of cancer caused by destruction of the mesenteric integrity may be the fundamental reason for the high postoperative recurrence rate. Using the nano carbon and indocyanine green fluorescence tracing technique, we demonstrated the lymphatic drainage of the upper esophageal segment to the left gastric artery mesenteric lymph nodes. Therefore, in the radical resection of esophageal cancer, we used the membrane anatomy theory for guidance to completely remove the esophageal cancer, esophageal mesentery, left gastric artery and its mesentery, as well as all structures within the mesentery, preventing the spread of cancer cells through the blood vessels, lymphatic system, and mesentery, and improving the efficacy and prognosis. This article elaborates on the theoretical basis of the anatomical structure of the esophageal membrane, embryonic development, imaging, autopsy, and endoscopic observation of the structure, as well as the application effect of the esophageal membrane anatomical theory in esophageal cancer radical surgery. It elucidates the anatomical structure of the esophageal membrane and the lymphatic drainage characteristics of esophageal cancer, reveals the law of lymphatic metastasis in esophageal cancer, optimizes lymphatic dissection strategies, and improves the efficacy of esophageal cancer radical surgery.

Transport of heterometric sediments in wave-dominated flows – Tracer experiments

Sediment dynamics is a complex subject, playing an important role in coastal areas. The interaction between wave action and sediment particles is determinant to understand sediment transport processes. The knowledge of sediment transport comprising sand mixtures is relevant as coastal areas, usually, present heterogeneities of sediment particle sizes in the horizontal and vertical directions, denoting the existence of selective transport processes. This study investigates the effects of non-cohesive size-graded sands on sediment transport fluxes to understand selective transport mechanisms under wave-dominated conditions. A laboratory experiment was carried out at the large wave flume (Großer Wellenkanal, GWK) in Hannover, considering distinct sediment mixtures between two well-sorted sands with median grain sizes of 0.21 mm and 0.58 mm. The experiments were carried out for four sediment bed configurations with two different wave energy conditions. A fluorescent sand tracer technique was implemented to measure total and fractional net sediment transport rates and the outcomes are compared with the ones obtained with the mass conservation method. The experimental results show the occurrence of the interaction between different sand fractions: a decrease in the transport of fine sediments and an enhancement of the transport of the coarse sediments as the percentage of coarse sediments in the bed mixture increases. These effects are larger as the coarser fraction in the sand bed increases and they are noticeable for different sediment transport regimes (sheet flow and rippled beds).

A novel air-assisted rotor spinning technique for ultra-stable antibacterial nanofiber/cotton hybrid yarn

Antibacterial textiles have attracted much attention in recent years. The stability of the antibacterial effect is one of the most important properties of antibacterial textiles. Integrating antibacterial nanofibers into cotton yarn is a green and efficient method to produce antibacterial textiles. However, due to the loose yarn structure of traditional rotor spinning, the functional nanofibers are easily detached from the blended yarns, resulting in reduced antibacterial effect. Herein, we modified the rotor spinning unit by adding an extra air supply channel for tighter yarn structure. The airflow field of the modified rotor spinning unit was simulated using computational fluid dynamics to determine the best setting angle of the extra air supply channel. Then, the antibacterial blended yarn was produced by trans-scale electrospinning and followed by air-assisted rotor spinning. At the same yarn density, the mean diameter of modified rotor spinning hybrid yarn was smaller than that of conventional rotor spinning hybrid yarn, demonstrating that the structure of modified rotor spinning hybrid yarn was tighter. The overall qualities of modified rotor spinning hybrid yarn were much better than those of conventional rotor spinning hybrid yarn. The fluorescent tracer technique was carried out to show that more nanofibers can be preserved in modified rotor spinning hybrid yarn than in conventional rotor spinning hybrid yarn, especially after 10 washing cycles. The antibacterial properties of modified rotor spinning hybrid yarn-based fabric against Escherichia coli and Staphylococcus aureus can reach as high as 80.5% and 82% even after 50 times of washing, indicating the high antibacterial durability. Our new technology provides a method to prepare super stable antibacterial functional yarn, and is expected to be used to prepare other durable functional textiles.

Effect of plasma treatment on the surface characteristics and adhesive penetration performance of heat-treated wood

Abstract The present study used an atmospheric dielectric barrier discharge (DBD) plasma treatment to modify the interfacial properties of heat-treated wood (HTW). The changes in the wood surface characteristics, such as the microstructure, chemical structure, and wettability, were investigated using scanning electron microscopy (SEM), mercury intrusion porosimetry (MIP), Fourier-transform infrared (FTIR) spectroscopy, and X-ray photoelectron spectroscopy (XPS). Further, the fluorescent tracer technique was used to analyze the effect of plasma treatment on the adhesive penetration inwood. Results showed that the wettability of water and phenol-formaldehyde adhesives on the surface of HTW significantly improved after plasma treatment due to the increase in the hydrophilic and oxygen-containing functional groups. The average penetration (AP) and effective penetration (EP) of the phenolic adhesive into HTW also increased significantly after plasma treatment; the AP value of the E-E (earlywood bound to earlywood) HTW samples increased from 909 to 1674 μm, and the EP value from 60 to 183 μm upon plasma treatment. These observations indicate that the DBD plasma treatment is a promising method to improve the bonding properties of the glued HTW products.

A Fluorescent Tracer Based on Castor Oil for Monitoring the Mass Transfer of Fatliquoring Agent in Leather.

Fatliquoring is one of the most important processes in leather making, in which the collagen fibers are split effectively, while the crust acquires a good softness and mechanical strength. The effectiveness of fatliquoring depends not only on the variety and dosage of fatliquoring agent but also on its distribution in hierarchical collagen fiber layers. Up to now, the research about the mass transfer of fatliquoring agent in leather is very limited because quantifying the distribution of invisible fatliquor in leather is very difficult. In this paper, a fluorescent tracing technique based on modified castor oil was established and send used to investigate the mass transfer of fatliquor in leather. The crucial fluorescent tracer was synthesized favorably by the reaction of castor oil, successively with maleic anhydride and 5-aminofluorescein, which was confirmed by FTIR, 1H NMR and DSC. The fluorescent tracer was pH-sensitive and emitted an intense fluorescent signal at pH 8–10. Then, it was applied to monitor the fatliquoring process in real-time. Compared with Sudan IV or Nile Blue sulphate dyeing tests, this fluorescent tracing technique could conveniently visualize and quantify the penetration and distribution of fatliquor in leather process.

Toxicokinetics of mono-(2-ethylhexyl) phthalate with low-dose exposure applying fluorescence tracing technique

Di(2-ethylhexyl) phthalate (DEHP) belongs to environmental endocrine disrupting chemicals (EEDCs) and can be rapidly hydrolyzed into the ultimate toxicant mono-2-ethylhexyl phthalate (MEHP). In this study, we used 5-aminofluorescein modified MEHP (MEHP-AF) as a fluorescence tracer to explore the toxicokinetics, including toxicokinetic parameters, absorption and transport across the intestinal mucosal barrier, distribution and pathological changes of organs. While the dose was as lower than 10 mg/kg by intragastric administration, the toxicokinetic parameters obtained by fluorescence microplate method were similar to those with the literatures by chromatography. MEHP-AF can be rapidly absorbed through the intestinal mucosal barrier in rats. In situ organ distribution in mice showed that MEHP-AF was mainly concentrated in the liver, kidney and testis. Our results suggested that the fluorescence tracing technique had the advantages with easy processing, less time-consuming, higher sensitivity for the quantitative determination, In addition, this technology also avoids the interference of exogenous or endogenous DEHP and MEHP in the experimental system. It also can be utilized to the visualization detection of MEHP in situ localization in the absorption organ and the toxic target organ. The results show that this may be a more feasible MEHP toxicological research method.

The Ethyl Acetate Extract From Celastrus orbiculatus Promotes Apoptosis of Gastric Cancer Cells Through Mitochondria Regulation by PHB.

Objective: To investigate the effect of ethyl acetate extract from Celastrus orbiculatus (COE) on gastric cancer cell apoptosis and reveal its underlying molecular mechanism. In addition, it was aimed to stablish a theoretical basis for the clinical application of Celastrus orbiculatus in the gastric cancer treatment. Material and Methods: Western blot and RT-qPCR were used to detect mRNA and protein expression of PHB in gastric cancer and adjacent tissues. MTT method was used to detect the COE effect on the proliferation of AGS cells and to determine the 50% inhibitory concentration COE on these cells. COE effect on AGS apoptosis was evaluated by flow cytometry. Changes in apoptosis-related proteins expression in AGS cells were detected by western blot and changes in mitochondrial membrane potential were detected by JC-1 fluorescence staining. PHB expression was knocked down in AGS cells by lentiviral-mediated RNA interference. The COE antitumor effect was assessed in vivo using a subcutaneous transplantation tumor model in nude mice and in vivo fluorescence tracing technique in small animals. Results: The clinical samples analysis results showed that the PHB expression in gastric cancer samples was significantly higher than in corresponding adjacent tissues. MTT results showed that the AGS cell proliferation was significantly inhibited. RT-qPCR and western blot results showed that COE can significantly inhibit the PHB mRNA and protein expression, respectively. Flow cytometry analysis showed that COE was able to significantly promote AGS cell apoptosis. Western blot results also indicated that apoptosis-related protein expression changed significantly; BCL-2 expression significantly reduced while the Caspase-3 and Bax expression significantly increased after COE treatment. JC-1 fluorescence staining results showed that COE changed the mitochondrial membrane potential and activated the mitochondrial apoptosis pathway. Furthermore, in vivo experiments results demonstrated that the growth of subcutaneous transplanted tumor was significantly inhibited by the PHB knockdown and by the COE intragastric administration. Conclusion: COE can significantly promote apoptosis of human gastric cancer cells, which can be achieved by inhibiting PHB expression, thus altering the structure and function of mitochondria and activating the mitochondria apoptosis pathway. The antitumor effect of COE has also been proved in vivo.

Correlated Sensory and Sympathetic Innervation Between the Acupoint BL23 and Kidney in the Rat.

Objective: To investigate the sensory and sympathetic innervations associated with both acupoint “Shenshu” (BL23) and kidney in the rat for insight into the neuronal correlation between the Back-Shu Point and its corresponding visceral organ.Methods: The BL23 and kidney were selected as the representative acupoint and visceral organ in this study, in which their local nerve fibers were examined by using double fluorescent immunohistochemistry with calcitonin gene-related peptide (CGRP) and tyrosine hydroxylase (TH). Meanwhile, their neuronal correlation in the dorsal root ganglia (DRGs), spinal cord, and sympathetic (paravertebral) chain were investigated using a double fluorescent neural tracing technique with Alexa Fluor 488 and 594 conjugates with cholera toxin subunit B (AF488/594-CTB).Results: The local tissue of acupoint BL23 and the fibrous capsule of kidney distributed abundantly with CGRP- and TH-positive nerve fibers, corresponding to their sensory and sympathetic innervation. On the other hand, the sensory neurons associated with acupoint BL23 and kidney were labeled with AF488/594-CTB and distributed from thoracic (T) 11 to lumbar (L) 3 DRGs and from T10 to L2 DRGs, respectively, in which some of them in T12-T13 DRGs were simultaneously labeled with both AF488/594-CTB. Also, postganglionic neurons associated with both acupoint BL23 and kidney were found in the sympathetic chain at the same spinal segments but separately labeled with AF488-CTB and AF594-CTB.Conclusion: Our study demonstrates the neural characteristics of the acupoint BL23 and kidney in the rat from the perspective of neurochemistry and neural pathways, providing an example for understanding the neuronal correlation between the Back-Shu Points and their corresponding visceral organs. These results suggest that the stimulation of the Back-Shu Points may regulate the activities of the target-organs via the periphery sensory and sympathetic pathways.

Interaction between retanning agents and wet white tanned by a novel bimetal complex tanning agent

A promising and practical chrome-free tanning system has been developed based on a novel Al–Zr bimetal complex tanning agent. However, to achieve satisfactory resultant leather, the retanning process that is compatible with this emerging tannage needs to be investigated systematically. This paper aims to explore the interaction between the bimetal complex tanned wet white and retanning agents. The isoelectric point (pI) of wet white was 7.2, which was nearly the same as wet blue. The electropositivity of wet white was even higher than that of wet blue during post-tanning processes, resulting in higher uptake rate of retanning agents. The distribution of various retanning agents in wet white was analyzed by pI measurement of layered leather and fluorescent tracing technique. The retanning agents were unevenly distributed throughout the cross-section, which might be an important restriction factor in obtaining satisfactory organoleptic properties of the crust leather. This fact is mainly due to the strong electrostatic interaction between anionic retanning agents and wet white. Applying a high dosage of multiple retanning agents in a proper sequence of addition benefited the full penetration of retanning agents in leather matrix and thus improved the organoleptic properties of crust leather. This work provides guidance for optimizing retanning process of the wet white leather.Graphical abstract

An efficient hybrid strategy for composite yarns of micro-/nano-fibers

In contrast to yarns of micro-fibers and those of nano-fibers, composite yarns composed of micro- and nano-fibers exhibit great potential in functional applications by integrating the respective advantages of multiscale fibers. However, the efficient fabrication of composite yarns remains challenging. So far, the existing nano-fiber included yarns, such as nanofiber yarn and core-sheath yarn, exhibit poor durability and low preparation efficiency. Herein, an efficient hybrid strategy for the composite yarns made of multiscale fibers has been developed by integration of electrospun nano-fibers and webbing of micro-fibers. Polyacrylonitrile nano-fiber/viscose micro-fiber composite yarns have been prepared massively. To explore structure of the resulting composite yarns, fluorescent tracer technique was adopted and revealed that nano-fibers are dispersed within composite yarn uniformly, while the structure of composite yarns can be governed easily by adjusting electrospinning parameters. Compared to pure yarns of viscose micro-fibers, the composite yarns possess mechanical properties at the same level, enabling the weaving, knitting and braiding of it for industrial applications. Furthermore, the formation mechanism of the composite yarn has been investigated and the fibrous adhesion among multiscale fibers has been modeled. The efficient approach should be a promising one for the design and scalable fabrication of composite yarns of micro-/nano-fibers.

Experimental Study of the Mixing and Segregation Behavior in Binary Particle Fluidized Bed with Wide Size Distributions

Abstract Detailed understanding the particle mixing and segregation dynamic is essential in successfully designing and reasonably operating multicomponent fluidized bed. In this work, a novel fluorescent tracer technique combining image processing method has been used to investigate the mixing and segregation behavior in a binary fluidized bed with wide size distributions. The particle number percentage in each layer for different gas velocities is obtained by an image processing method. Fluidization, mixing and segregation behavior has been discussed in terms of bed pressure drop, gas velocity and mixing index. Different types of binary particle systems, including the jetsam and the flotsam-rich system, are analyzed and compared. The mixing indexes at different minimum fluidization velocities are also analyzed and compared with other work. The results show that the theoretical minimum fluidization velocity calculated from the bed pressure drop cannot represent the whole fluidization for a wide size distribution binary particle system. The effect of a wide size distribution is an inflection point in the mixing index curve. There is also a dead region in the bottom of the bed that consists of particles with large size and a low degree of sphericity. The particles in the dead region are extraordinarily difficult to fluidize and should be considered in the design of fluidized beds in industrial applications.

Effects of Helicobacter pylori on the expression levels of GATA-3 and connexin 32 and the GJIC function in gastric epithelial cells and their association by promoter analysis.

The present study aimed to explore the effects of Helicobacter pylori (H. pylori) infection on the expression of transcription factor GATA binding protein 3 (GATA-3) and connexin 32 (Cx32) in cultured gastric mucosa cells, and their association with each other. GES-1 cells were co-cultured with East Asian type cytotoxin-associated gene A+ H. pylori in the H. pylori group, and without H. pylori culture in the control group. Additionally, Mongolian gerbils were gavaged with H. pylori, and later the gastric antrum tissues were collected. The GATA-3 and Cx32 mRNA and protein expression levels were detected by a reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. The scratch labeling fluorescent dye tracer (SLDT) technique was used to detect the gap junctional intercellular communication (GJIC) function. GATA-3 small interfering RNA (siRNA) was transfected into BGC823 cells and its effect on Cx32 expression levels was detected. The impact of GATA-3 on Cx32 promoter transcriptional activity was detected using a dual luciferase reporter assay. The results revealed that H. pylori infection increased GATA-3 expression and decreased Cx32 expression in GES-1 cells and in animal gastric tissues compared with their respective controls, whilst in BGC823 cells, GATA-3 siRNA increased Cx32 expression compared with the control. In the SLDT experiment of GES-1 cells with H. pylori infection, the fluorescent dye was primarily limited to a single cell row close to the scratch, and only a limited amount of dye passing to the second cell row, indicating that the GJIC function was substantially reduced or absent compared with the control group, where the fluorescence dye transferred to the neighboring cells of 3–4 rows, indicating a stronger GJIC function comparatively. GATA-3 inhibited the expression of the luciferase reporter gene, compared with the controls, suggesting that GATA-3 inhibited the expression of Cx32 by binding to Cx32 promoter sites. These results indicated that H. pylori-increased GATA-3 expression, which downregulated Cx32 expression, may serve an important function in gastric carcinogenesis, and GATA-3 siRNA may serve a function in the prevention and treatment of gastric cancer.

Microbiome Remodeling via the Montmorillonite Adsorption-Excretion Axis Prevents Obesity-related Metabolic Disorders

Obesity and its related metabolic disorders are closely correlated with gut dysbiosis. Montmorillonite is a common medicine used to treat diarrhea. We have previously found that dietary lipid adsorbent-montmorillonite (DLA-M) has an unexpected role in preventing obesity. The aim of this study was to further investigate whether DLA-M regulates intestinal absorption and gut microbiota to prevent obesity-related metabolic disorders. Here, we show that DLA-M absorbs free fatty acids (FFA) and endotoxins in vitro and in vivo. Moreover, the combination of fluorescent tracer technique and polarized light microscopy showed that DLA-M crystals immobilized BODIPY® FL C16 and FITC-LPS, respectively, in the digestive tract in situ. HFD-fed mice treated with DLA-M showed mild changes in the composition of the gut microbiota, particularly increases in short-chain fatty acids (SCFA)-producing Blautia bacteria and decreases in endotoxin-producing Desulfovibrio bacteria, these changes were positively correlated with obesity and inflammation. Our results indicated that DLA-M immobilizes FFA and endotoxins in the digestive tract via the adsorption-excretion axis and DLA-M may potentially be used as a prebiotic to prevent intestinal dysbiosis and obesity-associated metabolic disorders in obese individuals.

Assessment of pesticide application quality with a manual sprayer in spinach

Two experiments were conducted to evaluate the performance of three hollow cone nozzles (TeeJet® TX 800050 VK, TeeJet® TXA 8004 VK and TX-Royal Condor®) and one flat fan nozzle (TeeJet® XR 8004 VS) with two manual application techniques on a crop of spinach (Spinacia oleracea L.). In order to assess the quality ofthe application of pesticides, WSP (water sensitive paper) collectors and fluorescent tracer Tinopal® CBS-X were used. In one ofthe trials, percentage of tracer retained by the leaf surfaces was also determined. In this study, and based on the methodology of collectors, it was observed that the technique of applying two passes with the TX-Royal Condor® nozzle could be recommended for the application of pesticides with a hand-operated sprayer in the spinach crop. However, this was not corroborated by the fluorescent tracer technique.

Preliminary Analysis of Cellular Sociology of Co-Cultured Glioma Initiating Cells and Macrophages in Vitro

Objective Real-time monitoring of cytokine secretion at the single immunocyte level, based on the concept of immune cells, sociology has been recently reported. However, the relationships between glioma-initiating cells (GICs) and host immune cells and their mutual interactions in the tumor microenvironment have not been directly observed and remain unclear. Methods The dual fluorescence tracing technique was applied to label the co-cultured GICs and host macrophages (Mø), and the interactions between the two types of cells were observed using a live cell imaging system. Fusion cells in the co-culture system were monocloned and proliferated in vitro and their social interactions were observed and recorded. Results Using real-time dynamic observation of target cells, 6 types of intercellular conjunction microtubes were found to function in the transfer of intercellular information between GICs and Mo; GICs and host Mø can fuse into hybrid cells after several rounds of mutual interactions, and then these fusion cells fused with each other; Fusion cells generated offspring cells through symmetrical and asymmetrical division or underwent apoptosis. A “cell in cell” phenomenon was observed in the fusion cells, which was often followed by cell release, namely entosis. Conclusions Preliminary studies revealed the patterns of cell conjunction via microtubes between GICs and host Mø and the processes of cell fusion, division, and entosis. The results revealed malignant transformation of host Mø, induced by GICs, suggesting complex social relationships among tumor-immune cells in gliomas.

ATPS-18PRELIMINARY ANALYSIS ON SOCIAL ACTIVITIES OF CO-CULTURED GLIOMA STEM/PROGENITOR CELLS AND MACROPHAGES IN VITRO: CELL CONJUNCTION, CELL FUSION, AND ENTOSIS

BACKGROUND: The roles of real-time monitoring of cytokine secretion at the single immunocyte level, using the concept of immune cells sociology have been reported recently. However, the relationships between glioma stem/progenitors cells (GSPCs) and host immune cells and their interactions in tumor micro-environment have not been fully elucidated. METHODS: The dual fluorescence tracing technique were applied to label the co-cultured GSPCs and host macrophages, and the interactions between the two kinds of cells were observed under live cell imaging system. The yellow cells in the co-culture system were monocloned, and proliferated in vitro and then their social activities were observed and recorded too. RESULTS: Via real-time dynamic observation of the target cells, 1. there were 6 types of intercellular conjunction microtubes between GSPCs and macrophages functioned as transferring intercellular information; 2. GSPCs and host macrophages can fuse into hybrid cell after several rounds of mutual interactions, and one fusion cell could fuse with another fusion cell again; 3. Fusion cell could generate offspring cells through symmetrical division/asymmetrical division, or to undergo apoptosis; 4. One fusion cell in another fusion cell can be found, which can result in releasing out another cell, namely entosis. CONCLUSIONS: The preliminary studies revealed the patterns of cell conjunction between GSPCs and host macrophages, and the processes of cell fusion, division and entosis. These were cell sociological evidences of malignant transformation of host macrophages induced by GSPCs, which offered the basic knowledge exploring the complex tumor-immune cells relationships among the residents in the tumor cell society.