Experiment on inducing apoptosis of melanoma cells by micro-plasma jet

As a promising cancer treatment method,cold atmospheric plasma has received widespread attention in recent years.However,previous research has focused more on how to realize and expand the anti-cancer scope of plasma jet.There are also studies on the killing of small-scale cancer cells,but the effects of plasma jet on normal cells and normal cell clusters have been ignored.Therefore,we proposed a 50μm sized micro-plasma jet device,and used the device to treat melanoma cells(A-375)and human glial cells(HA1800)to evaluate their anti-cancer effects and effects on normal cells.The experimental results show that this kind of micro-plasma jet device can effectively inactivate cancer cells in a short period of time,while having little effect on normal cells.This work provides a certain experimental basis for the application offine plasma jet to clinically inactivate cancer cells.

Directional-to-random transition of cell cluster migration

Efficient cell migration is crucial for the functioning of biological processes, e.g., morphogenesis, wound healing, and cancer metastasis. In this study, we monitor the migratory behavior of the 3D fibroblast clusters using live cell microscopy,and find that crowded environment affects cell migration, i.e., crowding leads to directional migration at the cluster’s periphery. The number of cell layers being stacked during seeding determines the directional-to-random transition. Intriguingly,the migratory behavior of cell clusters resembles the dispersion dynamics of clouds of passive particles, indicating that the biological process is driven by physical effects(e.g., entropy) rather than cell communication. Our findings highlight the role of intrinsic physical characteristics, such as crowding, in regulating biological behavior, and suggest new therapeutic approaches targeting at cancer metastasis.

Advanced Coverage Optimization Techniques for Small Cell Clusters

Coverage challenge for small considered to be a optlmlzation is a main cell clusters which are promising solution to provide seamless cellular coverage for large indoor or outdoor areas. This paper focuses on small cell cluster coverage problems and proposes both centralized and distributed self-optimization methods. Modified Particle swarm optimization （MPSO） is introduced to centralized optimization which employs particle swarm optimization （PSO） and introduces a heuristic power control scheme to accelerate the algorithm to search tbr the global optimum solution. Distributed coverage optimization is modeled as a non-cooperative game, with a utility function considering both throughput and interference. An iterative power control algorithm is then proposed using game theory （DGT） which converges to Nash Equilibrium （NE）. Simulation results show that both MPSO and DGT have excellent performance in coverage optimization and outperform optimization using simulated annealing algorithm （SA）, reaching higher coverage ratio and throughput while with less iterations.

Effects of human umbilical cord serum on proliferation and insulin content of human fetal islet-like cell clusters

BACKGROUND: Type 1 diabets is an autoimmune disease caused by the destruction of pancreatic β-cell with an in- creased incidence worldwide in the closing decades of the 20th century. This study was to investigate the effects of human umbilical cord serum (UCS) on the proliferation and function of human fetal islet-like cell clusters (ICCs) in vitro. METHODS: Eight fresh pancreatic glands obtained after in- duction of labor with water bag were mildly exposed to col- lagenase V, and the digested cells were cultured in a RPMI- 1640 medium plus 10% pooled UCS or fetal calf serum (FCS) to permit cells attachment and outgrowth of ICCs. RESULTS: In 8 consecutively explanted glands, develop- ment and proliferation of ICCs were observed. In the pre- sence of FCS, the outgrowth of ICC took place on the top of a flbroblast monocellular layer. UCS affected less growth of fibroblasts and increased the formation of ICCs about four-fold compared with explants from the same glands maintained in FCS. In both UCS and FCS, the insulin con- tent of the medium was variable to a certain extent and progressively declined from day 2 to day 6. Dithizone- stained ICCs in UCS suggested that most cell clusters were islet cells ( β-cells), and the purity of islets was estimated 80%-90%. The ultrastructure of the cultured cells showed a large number of granule-containing cells, most of which were identified as β-cells. CONCLUSION: We conclude that in comparison with ex- plants with FCS, the yield of ICCs and purification of islet cells are markedly increased by UCS and may facilitate the proliferation of pancreatic β-cells intended for islet trans- plantation.

Algorithm to identify circulating tumor cell clusters using in vivo flow cytometer

Recent studies in oncology have addressed the importance of detecting circulating tumor cell clusters because circulating tumor cell clusters might survive and metastasize more easily than single circulating tumor cells.Signals with larger peak widths detected by in vivo flow cytometer(IVFC)have been used to identify cell clusters in previous studies.However,the accuracy of this criterion might be greatly degraded by variance in blood°ow and the rolling behaviors of circulating tumor cells.Here,we propose a criterion and algorithm to distinguish cell clusters from single cells.In this work,we first used area-based and volume-based models for single°uorescent cells.Simulating each model,we analyzed the corresponding morphology of IVFC signals from cell clusters.According to the Rayleigh criterion,the valley between two adjacent peak signals from two distinguishable cells should be lower than 73.5%of the peak values.A novel signal processing algorithm for IVFC was developed based on this criterion.The results showed that cell clusters can be reliably identied using our proposed algorithm.Intravital imaging was also performed to further support our algorithm.With enhanced accuracy,IVFC is a powerful tool to study circulating cell clusters.

Effects of EMS Treatments on Multiplication and Differentiation of Sugarcane Embryonic Cell Clusters

[ Objective] This study aimed to investigate the effects of EMS （ethyl methane sulfonate） treatments on multiplication and differentiation of sugarcane embryonic cell clusters. [ Method] Sugarcane variety Xintaitang 22 （ROC22） was used as the experimental material. After treated with different concentrations of EMS for different time, sugarcane embryonic cell clusters were collected for subculture, differentiation and rooting, to compare and analyze the correlation of differ- ent EMS treatments with the muhiplicafion and differentiation of sugarcane embryonic cell clusters. [ Result] Treating ROC22 embryonic cell clusters with 0. 10% -0.15% EMS for4-6 h led to the best results, which reached the level of semi-lethal dose. Sugarcane embryonic cell clusters treated with EMS were adopted for subculture; results indicated that browning rate of cell clusters was higher than that in control （CK） but embryonic structure proportion was lower than that in control ; in addition, multiplication multiple of sugarcane embryonic cell clusters was also lower than that in control. After treated with EMS, sugarcane em- bryonic cell clusters exhibited significantly lower bud differentiation rate and higher browning rate compared with control. Furthermore, treating sugarcane embryonic cell clusters with 0.15% EMS for 2 h was conducive to plantlet emergence and rooting of sugarcane embryonic cell clusters. [ Conclusion] This study provided the- oretical basis for effective mutagenesis of sugarcane using EMS.

Direct Differentiation of Plants from Small Cell Clusters of Indica Rice in Suspension Culture

The major factors affecting plant regeneration in suspension culture were investigated.The resultsshow that,in order for cell clusters to differentiate directly in liquid medium,it is essential toestablish an embryogenic callus line with high potential for plant regeneration.Embryogenic calliwere suspended in AA basic medium supplemented with 2,4-D 1 mg/L,6-benzylaminopurine 0.5mg/L,CH 300 mg/L,sucrose 3％ and mannitol 3％ and subcultured 7 days for each passage.Aftermore than 6 months of culturing,a fine suspension culture of small cell clusters(SCC)wasestablished.The SCC,80-270μin diameter,were then transferred to a liquid medium(MSsupplemented with NAA 0.01 mg/L and 4-pyridylurae(4-PU)0.5 mg/L)and allowed to grow instationary culture.Finally direct differentiation from SCC was observed.Several factors in suspension-subculture exerted strong after-effects on differentiation ofSCC.Basicmedia,kinds and concentration combinations of auxin and cytokinin in the subculture media,duration of shaking at each subculture passage,and amount of packed cell volume transferred to newmedium at each subculture passage and so on,all these affected the frequency of differentiation ofSCC.Higher concentrations of NAA and kinetin in the differentiation medium inhibited the directdifferentiation of SCC.Low concentrations(0.01-0.1 mg/L)of NAA with 4-PU in thedifferentiation medium were helpful for the direct differentiation of SCC.The differentiated clusterspossessed typical embryogenic structure from which normal plantlets could develop after transferringto agar medium.

A humanized mouse model to study human immune response in xenotransplantation

BACKGROUND: A major barrier to the clinical application of xenotransplantation as a treatment option for patients is T cell-mediated rejection. Studies based on experimental rodent models of xenograft tolerance or rejection in vivo have provided useful information about the role of T cell immune response in xenotransplantation. However not all observations seen in rodents faithfully recapitulate the human situation This study aimed to establish a humanized mouse model of xenotransplantation, which mimics xenograft rejection in the context of the human immune system. METHODS: NOD-SCID IL2rγ -/- mice were transplanted with neonatal porcine islet cell clusters (NICC) followed by reconstitution of human peripheral blood mononuclear cells (PBMC). Human leukocyte engraftment and islet xenograft rejection were confirmed by flow cytometric and histological analyses. RESULTS: In the absence of human PBMC, porcine NICC transplanted into NOD-SCID IL2rγ -/- mice revealed excellent graft integrity and endocrine function. Human PBMC demonstrated a high level of engraftment in NOD-SCID IL2rγ -/- mice. Reconstitution of NICC recipient NOD-SCID IL2rγ -/- mice with human PBMC led to the rapid destruction of NICC xenografts in a PBMC number-dependent manner. CONCLUSIONS: Human PBMC-reconstituted NOD-SCID IL2rγ -/- mice provide an ideal model to study human immune responses in xenotransplantation. Studies based on this humanized mouse model will provide insight for improving the outcomes of clinical xenotransplantation.

The model of electrified cell clusters in biological tissues basing on the resting potential difference

The resting potential is the potential difference that exists between the inner and outer sides of the cell membrane when the cell is not stimulated.The resting membrane potential is a key regulator of phenomena such as cell proliferation,morphogenesis,migration and differentiation.In this paper,we proposed a model of electrified cell clusters that considers the resting potential of cell clusters in the resting state.By measuring the potential difference between the inner and outer sides of biological tissues,it is verified that the cell cluster has a negative potential difference when taking the outer potential as the reference.In the absence of external conductors,the tissue is electrically neutral;while in the presence of external conductors,the positive net charges escape freely under repulsive forces and the biological tissues form a negative electrical equilibrium system.The model proposed in this study explored the potential situation above the cellular level in the resting state,providing a new perspective for the research on resting potential.

Cancer-specific calcium nanoregulator suppressing the generation and circulation of circulating tumor cell clusters for enhanced anti-metastasis combinational chemotherapy

Tumor metastasis is responsible for chemotherapeutic failure and cancer-related death.Moreover,circulating tumor cell(CTC)clusters play a pivotal role in tumor metastasis.Herein,we develop cancer-specific calcium nanoregulators to suppress the generation and circulation of CTC clusters by cancer membrane-coated digoxin(DIG)and doxorubicin(DOX)co-encapsulated PLGA nanoparticles(CPDDs).CPDDs could precisely target the homologous primary tumor cells and CTC clusters in blood and lymphatic circulation.Intriguingly,CPDDs induce the accumulation of intracellular Ca^(2+) by inhibiting Na^(+)/K^(+)-ATPase,which help restrain cellecell junctions to disaggregate CTC clusters.Meanwhile,CPDDs suppress the epithelialemesenchymal transition(EMT)process,resulting in inhibiting tumor cells escape from the primary site.Moreover,the combination of DOX and DIG at a mass ratio of 5:1 synergistically induces the apoptosis of tumor cells.In vitro and in vivo results demonstrate that CPDDs not only effectively inhibit the generation and circulation of CTC clusters,but also precisely target and eliminate primary tumors.Our findings present a novel approach for anti-metastasis combinational chemotherapy.