Artificial antigen-presenting cells plus IL-15 and IL-21 efficiently induce melanoma-specific cytotoxic CD8^+CD28^+ T lymphocyte responses

Objective:To develop a novel artificial antigen-presenting system for efficiently inducing melanoma-specific CD8+CD28+ cytotoxic T lymphocyte（CTL） responses.Methods:Cell-sized Dynabeads? M-450 Epoxy beads coated with H-2Kb:Ig-TRP2(181111K and anti-CD28 antibody were used as artificial antigen-presenting cells（aAPCs） lo induce melanoma-specific CD8*CD28’ CTL responses with the help of IL-2I and IL-I5.Dimer staining,proliferation,ELISPOT,and cytotoxicity experiments were conducted to evaluate the frequency and activity of induced CTLs.Results:Dimer staining demonstrated that the new artificial antigen-presenting system efficiently induced melanoma TRP2-specific CD8CD28' CTLs.Proliferation and ELISPOT assays indicated that the induced CTLs rapidly proliferate and produce increased IFN- y under the slimulalion of H-2K:Ig-TRP2-aAPCs,TL-15,and IL-21.In addition,cytoloxicily experiments showed lhat induced CTLs have specific killing activity of target cells.Conclusions:The new artificial antigen-presenting system including aAPCs plus IL-21 and IL-15 can induce a large number of antigen-specific CD8+CD28+ CTLs against the melanoma.Our study provides evidence for a novel adoptive immunotherapy against tumors.

Establishment and Characterization of a Cell Based Artificial Antigen-Presenting Cell for Expansion and Activation of CD8^+ T Cells Ex Vivo

Artificial antigen-presenting cells are expected to stimulate the expansion and acquisition of optimal therapeutic features of T cells before infusion. Here CD32 that binds to a crystallizable fragment of IgG monoclonal antibody was genetically expressed on human K562 leukemia cells to provide a ligand for T-cell receptor. CD86 and 4-1BBL, which are ligands of co-stimulating receptors of CD28 and 4-1BB, respectively, were also expressed on K562 cells. Then we accomplished the artificial antigen-presenting cells by coupling K32/CD86/4-1BBL cell with OKT3 monoclonal antibody against CD3, named K32/CD86/4-1BBL/OKT3 cells. These artificial modified cells had the abilities of inducing CD8^＋ T cell activation, promoting CD8^＋ T cell proliferation, division, and long-term growth, inhibiting CD8^＋ T cell apoptosis, and enhancing CD8^＋ T cell secretion of IFN-T and perforin. Furthermore, antigen-specific cytotoxic T lymphocytes could be retained in the culture stimulated with K32/CD86/4-1BBL/OKT3 cells at least within 28 days. This approach was robust, simple, reproducible and economical for expansion and activation of CD8^＋ T cells and may have important therapeutic implications for adoptive immunotherapy. Cellular ＆ Molecular Immunology.

A Novel and Functional Artificial Bruch’s Membrane Made of Poly (ε-Caprolactone)/Gelatin for Retinal Pigment Epithelium Restoration

Purpose: The primary aim of this paper is to design, create, and improve functional and artificial Bruch’s membranes (BM) using bioengineering techniques, which can be applied in the treatment of maculopathies by supporting the growth and maintenance of retinal pigment epithelium (RPE) cells, thereby potentially enabling subretinal implantation in patients. Methods: We fabricated by electrospinning ultrathin 3D nanofibrous membranes from Polycaprolactone (PCL), and different concentrations of gelatin (5%, 15% and 30%). ARPE-19 cells were seeded onto these artificial membranes. MTT assays were performed in order to evaluate ARPE-19 cell viability and cytotoxicity. IF assays were realized to observe the ARPE-19 cells onto each membrane. Ultrastructure of the modified Bruch’s membrane and ARPE-19 morphology after 25 days of culture were studied with transmission and scanning electron microscopy. To evaluate expression changes in markers of ARPE-19 (RPE65 and ZO-1) qRT-PCR assays were realized. Data from three independent experiments were pooled and expressed as the mean SD. A confidence level of P 0.05 was considered to be statistically significant. Results ARPE-19 cells grew on PCL/Gelatin membranes mainly in PCL/30% gelatin combination, which had not cytotoxic effect. RPE65/cytokeratin-18/ and actin-positive ARPE-19 cells formed a correctly orientated monolayer of polygonal cells with morphological polarity. The apical cell surfaces exhibited abundant protruding microfolds. Moreover, zones of polygonal border look as if ARPE-19 cells fused among, suggesting the presence of tight junctions. The expression of RPE65 and ZO-1 was unchanged. Conclusions: PCL/30% Gelatine membranes may imitate the natural BM to such extent that they support RPE-cells and exhibited RPE-like morphology. The engineering of a human RPE monolayer with these artificial BM, emulating the in vivo retina, arouse their potential subretinal implantation in patients with wet age-related macular degeneration (AMD) where there is a rupture of the Bruch’s membrane.

Over-expression of programmed death-ligand 1 and programmed death-1 on antigen-presenting cells as a predictor of organ dysfunction and mortality during early sepsis: a prospective cohort study

BACKGROUND:This study aimed to explore the changes of programmed death-ligand 1(PDL1)and programmed death-1(PD-1)expression on antigen-presenting cells(APCs)and evaluate their association with organ failure and mortality during early sepsis.METHODS:In total,40 healthy controls and 198 patients with sepsis were included in this study.Peripheral blood was collected within the first 24 h after the diagnosis of sepsis.The expression of PDL1 and PD-1 was determined on APCs,such as B cells,monocytes,and dendritic cells(DCs),by flow cytometry.Cytokines in plasma,such as interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),interleukin-4(IL-4),IL-6,IL-10,and IL-17A were determined by Luminex assay.RESULTS:PD-1 expression decreased significantly on B cells,monocytes,myeloid DCs(mDCs),and plasmacytoid DCs(pDCs)as the severity of sepsis increased.PD-1 expression was also markedly decreased in non-survivors compared with survivors.In contrast,PD-L1 expression was markedly higher on mDCs,pDCs,and monocytes in patients with sepsis than in healthy controls and in non-survivors than in survivors.The PD-L1 expression on APCs(monocytes and DCs)was weakly related to organ dysfunction and infl ammation.The area under the receiver operating characteristic curve(AUC)of the PD-1 percentage of monocytes(monocyte PD-1%)+APACHE II model(0.823)and monocyte PD-1%+SOFA model(0.816)had higher prognostic value than other parameters alone.Monocyte PD-1%was an independent risk factor for 28-day mortality.CONCLUSION:The severity of sepsis was correlated with PD-L1 or PD-1 over-expression on APCs.PD-L1 in monocytes and DCs was weakly correlated with infl ammation and organ dysfunction during early sepsis.The combination of SOFA or APACHE II scores with monocyte PD-1%could improve the prediction ability for mortality.

Artificial nerve graft constructed by coculture of activated Schwann cells and human hair keratin for repair of peripheral nerve defects

Studies have shown that human hair keratin(HHK) has no antigenicity and excellent mechanical properties. Schwann cells, as unique glial cells in the peripheral nervous system, can be induced by interleukin-1β to secrete nerve growth factor, which promotes neural regeneration. Therefore, HHK with Schwann cells may be a more effective approach to repair nerve defects than HHK without Schwann cells. In this study, we established an artificial nerve graft by loading an HHK skeleton with activated Schwann cells. We found that the longitudinal HHK microfilament structure provided adhesion medium, space and direction for Schwann cells, and promoted Schwann cell growth and nerve fiber regeneration. In addition, interleukin-1β not only activates Schwann cells, but also strengthens their activity and increases the expression of nerve growth factors. Activated Schwann cells activate macrophages, and activated macrophages secrete interleukin-1β, which maintains the activity of Schwann cells. Thus, a beneficial cycle forms and promotes nerve repair. Furthermore, our studies have found that the newly constructed artificial nerve graft promotes the improvements in nerve conduction function and motor function in rats with sciatic nerve injury, and increases the expression of nerve injury repair factors fibroblast growth factor 2 and human transforming growth factor B receptor 2. These findings suggest that this artificial nerve graft effectively repairs peripheral nerve injury.

Construction of Oriented Structure in Inner Surface of Small-Diameter Artificial Blood Vessels:A Review

There is an urgent need for small-diameter artificial blood vessels in clinic.Physical,chemical and biological factors should be integrated to avoid thrombosis and intimal hyperplasia after implantation and to promote successful fabrication of small-diameter artificial blood vessels.From a physical perspective,the internal oriented structures of natural blood vessels plays an important role in guiding the directional growth of cells,improving the blood flow environment,and promoting the regeneration of vascular tissue.In this review,the effects of the oriented structures on cells,including endothelial cells(ECs),smooth muscle cells(SMCs)and stem cells,as well as the effect of the oriented structures on hemodynamics and vascular tissue remodeling and regeneration are introduced.Various forms of oriented structures(fibers,grooves,channels,etc.)and their construction methods are also reviewed.Conclusions and future perspectives are given.It is expected to give some references to relevant researches.

Experimental study of bioartificial liver with cultured human liver cells

AIM To establish an extracorporeal bioartificial liver support system (EBLSS) using cultured human liver cells and to study its support effect for fulminant hepatic failure (FHF).METHODS The liver support experiment of EBLSS consisting of aggregates cultured human liver cells, hollow fiber bioreactor, and circulation unit was carried out in dizhepatic dogs.RESULTS The viability of isolated hepatocytes and nonparenchymal liver cells reached 96%. These cells were successfully cultured as multicellular spheroids with synthetic technique. The typical morphological appearance was retained up to the end of the artificial liver experiment. Compared with the control dogs treated with EBLSS without liver cells, the survival time of artificial liver support dogs was significantly prolonged. The changes of blood pressure, heart rate and ECG were slow. Both serum ammonia and lactate levels were significantly lowered at the 3rd h and 5th h. In addition, a good viability of human liver cells was noted after 5 h experiment.CONCLUSION EBLSS playing a metabolic role of cultured human hepatocytes, is capable of compensating the function of the liver, and could provide effective artificial liver support and therapy for patients with FHF.

Clinical translation of bioartificial liver support systems with human pluripotent stem cell-derived hepatic cells

There is currently a pressing need for alternative the-rapies to liver transplantation. The number of patients waiting for a liver transplant is substantially higher than the number of transplantable donor livers, resulting in a long waiting time and a high waiting list mortality. An extracorporeal liver support system is one possible approach to overcome this problem. However, the ideal cell source for developing bioartificial liver(BAL) support systems has yet to be determined. Recent advancements in stem cell technology allow researchers to generate highly functional hepatocyte-like cells from human pluripotent stem cells(h PSCs). In this mini-review, we summarize previous clinical trials with different BAL systems, and discuss advantages of and potential obstacles to utilizing h PSC-derived hepatic cells in clinical-scale BAL systems.

Artificial meat? Feasible approach based on the experience from cell culture studies

This short review is to list pros and cons which are based on the literature and personal experience in cell culture studies related to possible commercial production of artificial meat as functional food. The general view of muscle composition and determinants of meat quality are shortly described. Principles of muscle cell propagation in culture and mutual relationships between different cell types present in this organ are briefly discussed. Additionally, the effects of some cytokines and growth factors for muscle cell growth and muscle tissue development are indicated. Finally, conclusion remarks related to detrimental consequences of meat production to natural environment as well as personal opinion of author on the prospects of artificial meat production are declared.

Evaluation of corneal cell growth on tissue engineering materials as artificial cornea scaffolds

The keratoprosthesis(KPro;artificial cornea)is a special refractive device to replace human cornea by using heterogeneous forming materials for the implantation into the damaged eyes in order to obtain a certain vision.The main problems of artificial cornea are the biocompatibility and stability of the tissue particularly in penetrating keratoplasty.The current studies of tissue-engineered scaffold materials through comprising composites of natural and synthetic biopolymers together have developed a new way to artificial cornea.Although a wide agreement that the long-term stability of these devices would be greatly improved by the presence of cornea cells,modification of keratoprosthesis to support cornea cells remains elusive.Most of the studies on corneal substrate materials and surface modification of composites have tried to improve the growth and biocompatibility of cornea cells which can not only reduce the stimulus of heterogeneous materials,but also more importantly continuous and stable cornea cells can prevent the destruction of collagenase.The necrosis of stroma and spontaneous extrusion of the device,allow for maintenance of a precorneal tear layer,and play the role of ensuring a good optical surface and resisting bacterial infection.As a result,improvement in corneal cells has been the main aim of several recent investigations;some effort has focused on biomaterial for its well biological properties such as promoting the growth of cornea cells.The purpose of this review is to summary the growth status of the corneal cells after the implantation of several artificial corneas.

A method to deliver patterned electrical impulses to Schwann cells cultured on an artificial axon

Information from the brain travels back and forth along peripheral nerves in the form of electrical impulses generated by neurons and these impulses have repetitive patterns. Schwann cells in peripheral nerves receive molecular signals from axons to coordinate the process of myelination. There is evidence, however,that non-molecular signals play an important role in myelination in the form of patterned electrical impulses generated by neuronal activity. The role of patterned electrical impulses has been investigated in the literature using co-cultures of neurons and myelinating cells. The co-culturing method, however, prevents the uncoupling of the direct effect of patterned electrical impulses on myelinating cells from the indirect effect mediated by neurons. To uncouple these effects and focus on the direct response of Schwann cells,we developed an in vitro model where an electroconductive carbon fiber acts as an artificial axon. The fiber provides only the biophysical characteristics of an axon but does not contribute any molecular signaling.In our "suspended wire model", the carbon fiber is suspended in a liquid media supported by a 3D printed scaffold. Patterned electrical impulses are generated by an Arduino 101 microcontroller. In this study, we describe the technology needed to set-up and eventually replicate this model. We also report on our initial in vitro tests where we were able to document the adherence and ensheath of human Schwann cells to the carbon fiber in the presence of patterned electrical impulses(hSCs were purchased from ScienCell Research Laboratories, Carlsbad, CA, USA; ScienCell fulfills the ethic requirements, including donor's consent). This technology will likely make feasible to investigate the response of Schwann cells to patterned electrical impulses in the future.

Proton exchange membrane fuel cells modeling based on artificial neural networks

To understand the complexity of the mathematical models of a proton exchange membrane fuel cell (PEMFC) and their shortage of practical PEMFC control, the PEMFC complex mechanism and the existing PEMFC models are analyzed, and artificial neural networks based PEMFC modeling is advanced. The structure, algorithm, training and simulation of PEMFC modeling based on improved BP networks are given out in detail. The computer simulation and conducted experiment verify that this model is fast and accurate, and can be used as a suitable operational model for PEMFC real-time control.

Application of an artificial intelligence system for endoscopic diagnosis of superficial esophageal squamous cell carcinoma

BACKGROUND Upper gastrointestinal endoscopy is critical for esophageal squamous cell carcinoma(ESCC)detection;however,endoscopists require long-term training to avoid missing superficial lesions.AIM To develop a deep learning computer-assisted diagnosis(CAD)system for endoscopic detection of superficial ESCC and investigate its application value.METHODS We configured the CAD system for white-light and narrow-band imaging modes based on the YOLO v5 algorithm.A total of 4447 images from 837 patients and 1695 images from 323 patients were included in the training and testing datasets,respectively.Two experts and two non-expert endoscopists reviewed the testing dataset independently and with computer assistance.The diagnostic performance was evaluated in terms of the area under the receiver operating characteristic curve,accuracy,sensitivity,and specificity.RESULTS The area under the receiver operating characteristics curve,accuracy,sensitivity,and specificity of the CAD system were 0.982[95%confidence interval(CI):0.969-0.994],92.9%(95%CI:89.5%-95.2%),91.9%(95%CI:87.4%-94.9%),and 94.7%(95%CI:89.0%-97.6%),respectively.The accuracy of CAD was significantly higher than that of non-expert endoscopists(78.3%,P<0.001 compared with CAD)and comparable to that of expert endoscopists(91.0%,P=0.129 compared with CAD).After referring to the CAD results,the accuracy of the non-expert endoscopists significantly improved(88.2%vs 78.3%,P<0.001).Lesions with Paris classification type 0-IIb were more likely to be inaccurately identified by the CAD system.CONCLUSION The diagnostic performance of the CAD system is promising and may assist in improving detectability,particularly for inexperienced endoscopists.

Hunting down the dominating subclone of cancer stem cells as a potential new therapeutic target in multiple myeloma: An artificial intelligence perspective

The development of single-cell subclones,which can rapidly switch from dormant to dominant subclones,occur in the natural pathophysiology of multiple myeloma(MM)but is often"pressed"by the standard treatment of MM.These emerging subclones present a challenge,providing reservoirs for chemoresistant mutations.Technological advancement is required to track MM subclonal changes,as understanding MM's mechanism of evolution at the cellular level can prompt the development of new targeted ways of treating this disease.Current methods to study the evolution of subclones in MM rely on technologies capable of phenotypically and genotypically characterizing plasma cells,which include immunohistochemistry,flow cytometry,or cytogenetics.Still,all of these technologies may be limited by the sensitivity for picking up rare events.In contrast,more incisive methods such as RNA sequencing,comparative genomic hybridization,or whole-genome sequencing are not yet commonly used in clinical practice.Here we introduce the epidemiological diagnosis and prognosis of MM and review current methods for evaluating MM subclone evolution,such as minimal residual disease/multiparametric flow cytometry/next-generation sequencing,and their respective advantages and disadvantages.In addition,we propose our new single-cell method of evaluation to understand MM's mechanism of evolution at the molecular and cellular level and to prompt the development of new targeted ways of treating this disease,which has a broad prospect.

ARTIFICIAL IMMUNE ALGORITHM OF MULTICELLULAR GROUP AND ITS CONVERGENCE

Objective To find out more extrema simultaneously including global optimum and multiple local optima existed in multi-modal functions. Methods Germinal center is the generator and selector of high-affinity B cells, a multicellular group's artificial immune algorithm was proposed based on the germinal center reaction mechanism of natural immune systems. Main steps of the algorithm were given, including hyper-mutation, selection, memory, similarity suppression and recruitment of B cells and the convergence of it was proved. Results The algorithm has been tested to optimize various multi-modal functions, and the simulation results show that the artificial immune algorithm proposed here can find multiple extremum of these functions with lower computational cost. Conclusion The algorithm is valid and can converge on the satisfactory solution set D with probability 1 and approach to global solution and many local optimal solutions existed.

Cultivating human liver cell line (CL-1) on microcarriers as biomaterial of bioartificial liver

objective: To cultivate human liver cell line (CL-1) on microcarriers and study the synthetic and transformational function of this culture system. Methods:CL-1 were cultivated on Cytodex-3 microcarriers. The cell growth was kinetically inspected with light microscope and scanning electronic microscope on the lst, 3rd, 5th, 7th, 9th day, and the amount of diazepam transformation and albumin synthesis were deter mined at the same time. Results:On 7th day after inoculating, the CL-1 cell density could reach 2. 16 ×106/ ml ; the amount of diazepam trans formation was 619 μg and albumin synthesis 78. 23 μg. Conclusion:CL-1 can be cultivated to a high density on microcarriers and has hepatic specific biotransformation and biosynthesis functions. So the culture system may be further studied for being used as the biomaterial of bioartificial liver.

Application of artificial intelligence in clinical non-small cell lung cancer

Lung cancer is the most common cause of cancer death in the world.Early diagnosis,screening and precise individualized treatment can significantly reduce the death rate of lung cancer.Artificial intelligence(AI)has been shown to be able to help clinicians make more accurate judgments and decisions in many ways.It has been involved in the screening of lung cancer,the judgment of benign and malignant degree of pulmonary nodules,the classification of histological cancer,the differentiation of histological subtypes,the identification of genomics,the judgment of the effectiveness of treatment and even the prognosis.AI has shown that it can be an excellent assistant for clinicians.This paper reviews the application of AI in the field of non-small cell lung cancer and describes the relevant progress.Although most of the studies to evaluate the clinical application of AI in non-small cell lung cancer have not been repeatable and generalizable,the research results highlight the efforts to promote the clinical application of AI technology and influence the future treatment direction.

Recent trends in stem cell-based therapies and applications of artificial intelligence in regenerative medicine

Stem cells are undifferentiated cells that can self-renew and differentiate into diverse types of mature and functional cells while maintaining their original identity.This profound potential of stem cells has been thoroughly investigated for its significance in regenerative medicine and has laid the foundation for cellbased therapies.Regenerative medicine is rapidly progressing in healthcare with the prospect of repair and restoration of specific organs or tissue injuries or chronic disease conditions where the body’s regenerative process is not sufficient to heal.In this review,the recent advances in stem cell-based therapies in regenerative medicine are discussed,emphasizing mesenchymal stem cell-based therapies as these cells have been extensively studied for clinical use.Recent applications of artificial intelligence algorithms in stem cell-based therapies,their limitation,and future prospects are highlighted.

A fuzzy logic resource allocation and memory cell pruning based artificial immune recognition system

In order to improve the resource allocation mechanism of artificial immune recognition system(AIRS) and decrease the memory cells,a fuzzy logic resource allocation and memory cell pruning based AIRS(FPAIRS) is proposed.In FPAIRS,the fuzzy logic is determined by a parameter,thus,the optimal fuzzy logics for different problems can be located through changing the parameter value.At the same time,the memory cells of low fitness scores are pruned to improve the classifier.This classifier was compared with other classifiers on six UCI datasets classification performance.The results show that the accuracies reached by FPAIRS are higher than or comparable to the accuracies of other classifiers,and the memory cells decrease when compared with the memory cells of AIRS.The results show that the algorithm is a high-performance classifier.