Progress,implications,and challenges in using humanized immune system mice in CAR-T therapy-Application evaluation and improvement

In recent years,humanized immune system(HIS)mice have been gradually used as models for preclinical research in pharmacotherapies and cell therapies with major breakthroughs in tumor and other fields,better mimicking the human immune system and the tumor immune microenvironment,compared to traditional immunodeficient mice.To better promote the application of HIS mice in preclinical research,we se-lectively summarize the current prevalent and breakthrough research and evaluation of chimeric antigen receptor(CAR)-T cells in various antiviral and antitumor treat-ments.By exploring its application in preclinical research,we find that it can better reflect the actual clinical patient condition,with the advantages of providing high-efficiency detection indicators,even for progressive research and development.We believe that it has better clinical patient simulation and promotion for the updated design of CAR-T cell therapy than directly transplanted immunodeficient mice.The characteristics of the main models are proposed to improve the use defects of the existing models by reducing the limitation of antihost reaction,combining multiple models,and unifying sources and organoid substitution.Strategy study of relapse and toxicity after CAR-T treatment also provides more possibilities for application and development.

Skin care efficacy study of recombinant humanized collagen based on in vitro level

Studying the skin care efficacy of recombinant humanized collagen based on in vitro level.The stability of the recombinant humanized collagen was first analyzed by treating at different temperatures,then its skincare efficacy based on in vitro level was evaluated by detecting the inhibition rate of elastase,the inhibition rate of collagenase,the protein content of type I collagen in human fibroblasts,the inhibition of reactive oxygen species(ROS)with human keratinocytes,and the effects of the recombinant humanized collagen on the expression of hyaluronic acid(HA),filaggrin(FLG)and transglutaminase 1(TGM1)in keratinocytes.The results showed that recombinant humanized collagen was able to maintain stability at temperatures below 70℃.With regard to its skincare efficacy,recombinant humanized collagen could inhibit elastase and collagenase activities and promote the increase of type I collagen content in human fibroblasts.It also showed good inhibition of ROS in keratinocytes in vitro and could increase the expression of HA,FLG,and TGM1 in keratinocytes.In short,the recombinant humanized collagen exhibited a favourable skin care effect in vitro level.This study proved that it has potential firming,anti-wrinkle,moisturizing,and repairing efficacy,and is a valuable cosmetic raw material.

Establishment of a humanized ST6GAL1 mouse model for influenza research

Background:This study aimed to construct and characterize a humanized influenza mouse model expressing hST6GAL1.Methods:Humanized fragments,consisting of the endothelial cell-specific K18 promoter,human ST6GAL1-encoding gene,and luciferase gene,were microinjected into the fertilized eggs of mice.The manipulated embryos were transferred into the oviducts of pseudopregnant female mice.The offspring were identified using PCR.Mice exhibiting elevated expression of the hST6GAL1 gene were selectively bred for propagation,and in vivo analysis was performed for screening.Expression of the humanized gene was tested by performing immunohistochemical(IHC)analysis.Hematologic and biochemical analyses using the whole blood and serum of humanized hST6GAL1 mice were performed.Results:Successful integration of the human ST6GAL1 gene into the mouse genome led to the overexpression of human SiaT ST6GAL1.Seven mice were identified as carrying copies of the humanized gene,and the in vivo analysis indicated that hST6GAL1gene expression in positive mice mirrored influenza virus infection characteristics.The IHC results revealed that hST6GAL1 was expressed in the lungs of humanized mice.Moreover,the hematologic and biochemical parameters of the positive mice were within the normal range.Conclusion:A humanized influenza mouse model expressing the hST6GAL1 gene was successfully established and characterized.

Operation room nursing based on humanized nursing mode combined with nitric oxide on rehabilitation effect after lung surgery

BACKGROUND With advancements in the diagnosis and treatment of lung diseases,lung segment surgery has become increasingly common.Postoperative rehabilitation is critical for patient recovery,yet challenges such as complications and adverse outcomes persist.Incorporating humanized nursing modes and novel treatments like nitric oxide inhalation may enhance recovery and reduce postoperative complications.AIM To evaluate the effects of a humanized nursing mode combined with nitric oxide inhalation on the rehabilitation outcomes of patients undergoing lung surgery,focusing on pulmonary function,recovery speed,and overall treatment costs.METHODS A total of 79 patients who underwent lung surgery at a tertiary hospital from March 2021 to December 2021 were divided into a control group(n=39)receiving a routine nursing program and an experimental group(n=40)receiving additional humanized nursing interventions and atomized inhalation of nitric oxide.Key indicators were compared between the two groups alongside an analysis of treatment costs.RESULTS The experimental group demonstrated significant improvements in pulmonary function,reduced average recovery time,and lower total treatment costs compared to the control group.Moreover,the quality of life in the experimental group was significantly better in the 3 months post-surgery,indicating a more effective rehabilitation process.CONCLUSION The combination of humanized nursing mode and nitric oxide inhalation in postoperative care for lung surgery patients significantly enhances pulmonary rehabilitation outcomes,accelerates recovery,and reduces economic burden.This approach offers a promising reference for improving patient care and rehabilitation efficiency following lung surgery.

Research on the Application Effect of Humanized Nursing Management in Pediatric Rehabilitation Wards

Objective:To explore the application effect of humanized nursing management in pediatric rehabilitation wards.Methods:82 pediatric patients admitted to our hospital between January 2023 and January 2024 were randomly divided into a control group and an observation group.Among them,the control group was given routine care,while the observation group received humanized nursing management,and the clinical nursing effects and situations of the two groups of children were compared and analyzed.Results:After nursing,the observation group showed significantly better performance in intelligent Developmental Quotient(DQ)values(P<0.05).The clinical nursing satisfaction of the observation group was significantly better than that of the control group(χ^(2)=11.710,P<0.05).Conclusion:Humanized nursing management for patients in pediatric rehabilitation wards not only improves their recovery but also significantly enhances parental satisfaction,which is worth learning from and promoting in clinical settings.

Analysis of the Effect of Implementing Humanized Care Service in Severe ICU Patients

Objective:To analyze the effect of implementing humanized nursing service intervention for severe patients in the intensive care unit(ICU).Methods:A hundred severely ill ICU patients who were treated from January 2021 to December 2022 were selected and grouped into a control group and an observation group.The control group adopted routine nursing services and the observation group adopted humanized nursing services.The nursing outcome of the two groups was analyzed.Results:The nursing risk incidence of the observation group was lower than that of the control group(P<0.05).The scale of comfort and nursing satisfaction in the observation group was higher than those in the control group(P<0.05).Conclusion:The implementation of a humanized care service for ICU patients lowered nursing risk incidences and increased the physical comfort and nursing satisfaction of these patients.

Application and Satisfaction Analysis of Humanized Quality Nursing Intervention in the Care of Patients with Critical Respiratory Failure

Objective:To implement humanized quality care in critical respiratory failure nursing,observe the effect,and analyze the satisfaction.Methods:80 patients with severe respiratory failure were divided into 40 cases per group(admitted from February 2022 to December 2023)by double-blind method,the control group performed routine care,and the observation group was given humanized quality care.Results:After the nursing care,two respiratory function indexes of the observation group were lower than those of the control group,and two pulmonary function indexes were higher than those of the control group(P<0.05);regarding the complication situation,the incidence rate of the observation group was lower(P<0.05);regarding the nursing care satisfaction situation,the observation group had a higher level of total satisfaction(P<0.05).Conclusion:The application of a humanized quality nursing intervention model in the care of critical respiratory failure can actively improve patients’respiratory status and lung function,reduce complications,and satisfy patients.

Effects of humanized nursing care on negative emotions and complications in patients undergoing hysteromyoma surgery

BACKGROUND Uterine fibroids,are prevalent benign tumors affecting women of reproductive age.However,surgical treatment is often necessary for symptomatic hysteromyoma cases.This study examines the impact of humanized nursing care on reducing negative emotions and postoperative complications in patients receiving hysteromyoma surgery.AIM To investigate the impact of humanized nursing care on patients undergoing hysteromyoma surgery.METHODS Among patients who underwent hysteromyoma surgery at the Fudan University Affiliated Obstetrics and Gynecology Hospital,200 were randomly assigned to either the control group(n=100)or the humanized nursing care group(n=100).The control group received traditional nursing care,while the humanized nursing care group received a comprehensive care plan encompassing psychological support,pain management,and tailored rehabilitation programs.In addition,anxiety and depression levels were assessed using the hospital anxiety and depression scale preoperatively and postoperatively.Postoperative complications were evaluated during follow-up assessments and compared between both groups.RESULTS The humanized nursing care group demonstrated a significant decrease in anxiety and depression levels compared to the control group(P<0.05).The rate of postoperative complications,including infection,bleeding,and deep venous thrombosis,was also markedly lower in the humanized nursing care group(P<0.05).CONCLUSION Humanized nursing care can effectively alleviate negative emotions and reduce the incidence of postoperative complications in patients undergoing hysteromyoma surgery.This approach should be considered a crucial component of perioperative care for these patients.Further research may be needed to explore additional benefits and long-term outcomes of implementing humanized nursing care in this population.

Characterization of genetic humanized mice with transgenic HLA DP401 or DRA but deficient in endogenous murine MHC classⅡgenes upon Staphylococcus aureus pneumonia

Background:Staphylococcus aureus can cause serious infections by secreting many superantigen exotoxins in“carrier”or“pathogenic”states.HLA DQ and HLA DR humanized mice have been used as a small animal model to study the role of two molecules during S.aureus infection.However,the contribution of HLA DP to S.aureus infection is unknown yet.Methods:In this study,we have produced HLA DP401 and HLA DRA0101 humanized mice by microinjection of C57BL/6J zygotes.Neo-floxed IAβ+/-mice were crossbred with Ella-Cre and further crossbred with HLA DP401 or HLA-DRA0101 humanized mice.After several rounds of traditional crossbreeding,we finally obtained HLA DP401-IAβ-/-and HLA DRA-IAβ-/-humanized mice,in which human DP401 or DRA0101 molecule was introduced into IAβ-/-mice deficient in endogenous murine MHC classⅡmolecules.A transnasal infection murine model of S.aureus pneumonia was induced in the humanized mice by administering 2×108CFU of S.aureus Newman dropwise into the nasal cavity.The immune responses and histopathology changes were further assessed in lungs in these infected mice.Results:We evaluated the local and systemic effects of S.aureus delivered intranasally in HLA DP401-IAβ-/-and HLA DRA-IAβ-/-transgenic mice.S.aureus Newman infection significantly increased the m RNA level of IL 12p40 in lungs in humanized mice.An increase in IFN-γand IL-6 protein was observed in HLA DRA-IAβ-/-mice.We observed a declining trend in the percentage of F4/80+macrophages in lungs in HLA DP401-IAβ-/-mice and a decreasing ratio of CD4+to CD8+T cells in lungs in IAβ-/-mice and HLA DP401-IAβ-/-mice.A decreasing ratio of Vβ3+to Vβ8+T cells was also found in the lymph node of IAβ-/-mice and HLA DP401-IAβ-/-mice.S.aureus Newman infection resulted in a weaker pathological injury in lungs in IAβ-/-genetic background mice.Conclusion:These humanized mice will be an invaluable mouse model to resolve the pathological mechanism of S.aureus pneumonia and study what role DP molecule plays in S.aureus infection.

Development of a humanized HLA-A30 transgenic mouse model

Background:There are remarkable genetic differences between animal major histocompatibility complex(MHC)systems and the human leukocyte antigen(HLA)system.HLA transgenic humanized mouse model systems offer a much better method to study the HLA-A-related principal mechanisms for vaccine development and HLA-Arestricted responses against infection in human.Methods:A recombinant gene encoding the chimeric HLA-A30 monochain was constructed.This HHD molecule contains the following:α1-α2 domains of HLA-A30,α3 and cytoplasmic domains of H-2D~b,linked at its N-terminus to the C-terminus of humanβ2m by a 15-amino-acid peptide linker.The recombinant gene encoding the chimeric HLA-A30 monochain cassette was introduced into bacterial artificial chromosome(BAC)CH502-67J3 containing the HLA-A01 gene locus by Red-mediated homologous recombination.Modified BAC CH502-67J3 was microinjected into the pronuclei of wild-type mouse oocytes.This humanized mouse model was further used to assess the immune responses against influenza A virus(H1N1)pdm09 clinically isolated from human patients.Immune cell population,cytokine production,and histopathology in the lung were analyzed.Results:We describe a novel humanβ2m-HLA-A30(α1α2)-H-2D~b(α3 transmembrane cytoplasmic)(HHD)monochain transgenic mouse strain,which contains the intact HLA-A01 gene locus including 49 kb 5’-UTR and 74 kb 3’-UTR of HLA-A01*01.Five transgenic lines integrated into the large genomic region of HLA-A gene locus were obtained,and the robust expression of exogenous transgene was detected in various tissues from A30-18#and A30-19#lines encompassing the intact flanking sequences.Flow cytometry revealed that the introduction of a large genomic region in HLA-A gene locus can influence the immune cell constitution in humanized mice.Pdm09 infection caused a similar immune response among HLA-A30 Tg humanized mice and wild-type mice,and induced the rapid increase of cytokines,including IFN-γ,TNF-α,and IL-6,in both HLA-A30 humanized Tg mice and wild-type mice.The expression of HLA-A30 transgene was dramatically promoted in tissues from A30-9#line at 3 days post-infection(dpi).Conclusions:We established a promising preclinical research animal model of HLA-A30 Tg humanized mouse,which could accelerate the identification of novel HLA-A30-restricted epitopes and vaccine development,and support the study of HLA-A-restricted responses against infection in humans.

Generation and expression analysis of BAC humanized mice carrying HLA-DP401 haplotype

Background:Human leukocyte antigen(HLA)-DP is much less studied than other HLA class Ⅱ antigens,that is,HLA-DR and HLA-DQ,etc.However,the accumulating data have suggested the important roles of DP-restricted responses in the context of cancer,allergy,and infectious disease.Lack of animal models expressing these genes as authentic cis-haplotypes blocks our understanding for the role of HLA-DP haplotypes in immunity.Methods:To explore the potential cis-acting control elements involved in the tran-scriptional regulation of the HLA-DPA1/DPB1 gene,we performed the expression analysis using bacterial artificial chromosome(BAC)-based transgenic humanized mice in the C57BL/6 background,which carried the entire HLA-DP401 gene locus.We further developed a mouse model of Staphylococcus aureus pneumonia in HLA-DP401 humanized transgenic mice,and performed the analysis on the expres-sion pattern of HLA-DP401 and immunological responses in the model.Results:In this study,we screened and identified a BAC clone spanning the entire HLA-DP gene locus.DNA from this clone was analyzed for integrity by pulsed-field gel electrophoresis and then microinjected into fertilized mouse oocytes to produce transgenic founder animals.Nine sets of PCR primers for regional markers with an average distance of 15 kb between each primer were used to confirm the integrity of the transgene in the five transgenic lines carrying the HLA-DPA1/DPB1 gene.Transgene copy numbers were determined by real-time PCR analysis.HLA-DP401 gene expression was analyzed at the mRNA and protein level.Although infection with S aureus Newman did not alter the percentage of immune cells in the spleen and thymus from the HLA-DP401-H2-Aβ1 humanized mice.Increased expression of HLA-DP401 was observed in the thymus of the humanized mice infected by S aureus.Conclusions:We generated several BAC transgenic mice,and analyzed the expres-sion of HLA-DPA1/DPB1 in those mice.A model of S aureus-induced pneumonia in the HLA-DP401-H2-Aβ1^(-/-)humanized mice was further developed,and S aureus in-fection upregulated the HLA-DP401 expression in thymus of those humanized mice.These findings demonstrate the potential of those HLA-DPA1/DPB1 transgenic humanized mice for developing animal models of infectious diseases and MHC-associated immunological diseases.

The Effects of Humanized Psychological Nursing Model in General Surgery Nursing

Objective:To analyze and comprehensively study the clinical effects of humanized psychological nursing model in general surgery nursing.Methods:The study period was from January 2018 to December 2020.A sample of 200 patients who were admitted to The Second People's Hospital of Taizhou City for general surgery were selected.Random lottery grouping was used to divide the subjects into a study group and a control group.The sample within each group was n=100.The patients in the control group were provided with conventional general surgery nursing plan whereas the patients in the study group received the same nursing plan but with addition of the humanized psychological nursing model.The indicators of the two groups were compared and analyzed.Results:Comparing the scores from Hamilton Anxiety Rating Scale(HAMA)and Hamilton Depression Rating Scale(HAMD)after nursing intervention,postoperative visual analog scale(VAS),hospital stay,and patient satisfaction with the nursing services between the two groups,the study group was better(P<0.05).Conclusion:The implementation of humanized psychological nursing model in general surgery nursing had a significant effect in which there were improvements in regard to the patientsJ mental state and their satisfaction with the nursing services.Hence,it is worthy of promotion.

The Application of Landscape Plants in Humanized Interior Environmental Design

Landscape plants play a role of decorating and beautifying the indoor environment.From the perspective of humanization,the functions and application patterns of landscape plants in the interior environmental design are analyzed in this study,and finally the application rules of landscape plants in the humanized interior environmental design are summarized.

Bioengineered humanized livers as better three-dimensional drug testing model system

AIM To develop appropriate humanized three-dimensional ex-vivo model system for drug testing. METHODS Bioengineered humanized livers were developed in this study using human hepatic stem cells repopulation within the acellularized liver scaffolds which mimics with the natural organ anatomy and physiology. Six cytochrome P-450 probes were used to enable efficient identification of drug metabolism in bioengineered humanized livers. The drug metabolism study in bioengineered livers was evaluated to identify the absorption, distribution, metabolism, excretion and toxicity responses.RESULTS The bioengineered humanized livers showed cellular and molecular characteristics of human livers. The bioengineered liver showed three-dimensional natural architecture with intact vasculature and extra-cellular matrix. Human hepatic cells were engrafted similar to the human liver. Drug metabolism studies provided a suitable platform alternative to available ex-vivo and in vivo models for identifying cellular and molecular dynamics of pharmacological drugs.CONCLUSION The present study paves a way towards the development of suitable humanized preclinical model systems for pharmacological testing. This approach may reduce the cost and time duration of preclinical drug testing and further overcomes on the anatomical and physiological variations in xenogeneic systems.

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.

Feasibility study on construction of humanized mice with antibiotics combined with fecal bacteria transplantation

Objective:To select a common method of humanized mice and verify its feasibility with the aim to provide atheoretical basis for subsequent fecal bacteria transplantation research.Methods:1.Specific Pathogen Free(SPF)mice were fed with antibiotics mixed with sterilized drinking water for 3 weeks,during which the general situation of mice was recorded.After 3 weeks,the number of microbes in mice's feces was evaluated by Gram staining.2.After antibiotic treatment,mice were transplanted fecal bacteria by gavage for a week.Then,the contents of fecal bacteria in the donor and recipient feces was determined by RT-PCR,to evaluate the colonization effect of fecal bacteria.Results:(1)Antibiotic treatment had an effect on weight,feed and drinking water consumption of mice,and the fecal bacteria in mice decreased obviously.(2)After fecal bacteria transplantation,the Firmicutes,Bacteroidetes and Firmicutes/Bacteroidetes of the mice receiving Donor1 fecal bacteria accounted for 15.4%,112%and 13.7%of the mice receiving Donor2 fecal bacteria,respectively.The Firmicutes,Bacteroidetes and Firmicutes/Bacteroidetes of Donor1 fecal bacteria accounted for 33.7%,104%and 32%of Donor2,respectively.The recipient's gut microbes had the same trend as human donors.Conclusions:These results indicate that antibiotic treatment combined with fecal bacterial transplantation is feasible and safe for humanizing SPF mice.

Humanized Spatial Layout Design of Cultural Plaza

From the perspective of humanized design concepts, humanized space design and behavioral psychological needs of users were re-interpreted and analyzed, basic elements and design principles of cultural plaza spatial layout were proposed. Particularly, application and role of humanized design in the spatial layout of cultural plaza was analyzed, on the basis of which humanized design objective and orientation for the sustainable development of cultural plaza was defined.

Analysis on Application of Humanized Design Thinking in Ceramic Modeling

With the continuous development of human civilization, modem design has also made great progress. The design thinking tends more to adapting to human self-demand, in order to achieve the purpose of serving for people＇ s daily life. Therefore, the humanized design concept of modem society is proposed, and in many fields, the humanized design thinking has become mature. This paper mainly takes systematic and overall analysis of the application of humanized design thinking in ceramic modeling, in order to provide relevant reference for the majority of peers.

Research on New University Education Management Methods based on the Combination of Humanized and Institutionalized

In this paper, we conduct research on new university education management methods based on the combination of humanized and institutionalized. Modem university education management, system management and the organic integration of humanistic care is under the new period of development, university education management to sustainable development as the main direction of the management. System and manage all exist, man and man is the subject and purpose of system and management. The construction of system, arrangement and operation shall be at the center of the reality of human and highlight the cultural characteristics of follow the characteristic of the knowledge in the differentiation and specialization.

S4B-4 A Novel Allosteric Phosphodiesterase 4D Inhibitor BPN14770 Reverses Cognitive Impairment in Humanized PDE4D Mice

Classical inhibitors of PDE4 lack subtype selectivity due to exact amino acid sequence conservation of the catalytic site,and consequently,development of these drugs has stalled due to dose-limiting side effects of nausea and emesis.While use of subtype-selective inhibitors(i.e.,for PDE4A,B,or D)could overcome this issue,conservation of the catalytic region,to which classical inhibitors bind,limits this approach.The present study examined the effects of BPN14770,an allosteric inhibitor of PDE4D,which binds to a primate-specific,N-terminal region,conferring greater than 260-fold selectivity for PDE4D.BPN14770 was 100-fold more potent for improving memory and cognition in humanized PDE4D(hPDE4D)mice,which expressed the primate-specific binding sequence,compared to wild-type mice;meanwhile,it exhibited low potency in a mouse surrogate model for emesis.The behavioral and matching neurochemical data presented established a relationship between PDE4D target engagement and effects on cognition for BPN14770.Furthermore,BPN14770 reversed memory and cognitive deficits induced byβ-amyloid peptide 1-42(Aβ42)in Morris water maze,Y maze and novel object recognition tests in the humanized PDE4D mice.The morphological analyses suggested that the number of dendrites and the dendritic length in the CA1 of hippocampus were significantly increased after the Aβ42-treated hPDE4D mice were administered of BPN14770 for two weeks.The neurochemical and molecular biological assays suggested that neuroplasticity-related proteins and neurotrophic factor BDNF in the hippocampus of hPDE4D mice were significantly increased after the hPDE4D mice were treated with BPN14770.These findings suggest clinical potential for PDE4D selective inhibitors in disorders with cognitive deficits such as Alzheimer’s disease,which affects approximately 20 million people worldwide and nearly 5 million people in the United States.