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Hypoxia often occurs in inflammatory tissues, such as tissues affected by periodontitis and apical periodontitis lesions. Mitochondrial biogenesis can be disrupted in hypoxia. Peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) is a core factor required for mitochondrial biogenesis. Cementoblasts are root surface lining cells that play an integral role in cementum formation. There is a dearth of research on the effect of hypoxia on cementoblasts and underlying mechanisms, particularly in relation to mitochondrial biogenesis during the hypoxic process. In this study, we found that the expression of hypoxia inducible factor-1α was elevated in apical periodontitis tissues in vivo. In contrast, periapical lesions exhibited a reduction of PGC-1α expression. For in vitro experiments, cobalt chloride (CoCl

Brassica napus is an important oil crop and an allotetraploid species. However, the detailed analysis of gene function and homoeologous gene expression in all tissues at different developmental stages was not explored. In this study, we performed a global transcriptome analysis of 24 vegetative and reproductive tissues at six developmental stages (totally 111 tissues). These samples were clustered into eight groups. The gene functions of silique pericarp were similar to roots, stems and leaves. In particular, glucosinolate metabolic process was associated with root and silique pericarp. Genes involved in protein phosphorylation were often associated with stamen, anther and the early developmental stage of seeds. Transcription factor (TF) genes were more specific than structural genes. A total of 17 100 genes that were preferentially expressed in one tissue (tissue-preferred genes, TPGs), including 889 TFs (5.2%), were identified in the 24 tissues. Some TPGs were identified as hub genes in the co-expression network analysis, and some TPGs in different tissues were involved in different hormone pathways. About 67.0% of the homoeologs showed balanced expression, whereas biased expression of homoeologs was associated with structural divergence. In addition, the spatiotemporal expression of homoeologs was related to the presence of transposable elements (TEs) and regulatory elements (REs); more TEs and fewer REs in the promoters resulted in divergent expression in different tissues. This study provides a valuable transcriptional map for understanding the growth and development of B. napus, for identifying important genes for future crop improvement, and for exploring gene expression patterns in the B. napus.

Unlike PIWI-interacting RNA (piRNA) in other species that mostly target transposable elements (TEs), >80% of piRNAs in adult mammalian testes lack obvious targets. However, mammalian piRNA sequences and piRNA-producing loci evolve more rapidly than the rest of the genome for unknown reasons. Here, through comparative studies of chickens, ducks, mice, and humans, as well as long-read nanopore sequencing on diverse chicken breeds, we find that piRNA loci across amniotes experience: (1) a high local mutation rate of structural variations (SVs, mutations ≥ 50 bp in size); (2) positive selection to suppress young and actively mobilizing TEs commencing at the pachytene stage of meiosis during germ cell development; and (3) negative selection to purge deleterious SV hotspots. Our results indicate that genetic instability at pachytene piRNA loci, while producing certain pathogenic SVs, also protects genome integrity against TE mobilization by driving the formation of rapid-evolving piRNA sequences.

Cementum regeneration was regarded as the critical goal for periodontal regeneration, and M2 macrophage-based therapy was expected to be a promising strategy. However, little is known about the effects of M2 macrophages on cementoblast mineralization and tropism, especially under inflammation. Here we investigated for the first time the crosstalk between M2 macrophages and Porphyromonas gingivalis (Pg)-stimulated cementoblasts.M2 macrophages were induced with interleukin (IL)-4, and identified. CC-chemokine ligand 2 (CCL2) expression and secretion of inflammatory cementoblasts were detected by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), western blotting (WB), immunohistochemistry for apical periodontitis (AP) mice, and by enzyme-linked immunosorbent assay. Crystal violet staining was used to observe macrophage migration. Conditional medium (CM) and transwell coculture methods were applied to evaluate the effects of M2 macrophages on cementum mineralization with or without Pg, and to explore the mechanism. Mineralization-related markers and pathway-related proteins were measured by RT-qPCR and WB.M2 macrophages were identified successfully. We found an increase of CCL2 in cementoblasts and their supernatant. Also, higher CCL2 in cementoblasts was observed in the AP model. Superior recruitment of M2 macrophages to supernatant from Pg-stimulated cementoblasts or CCL2-containing medium was verified. Moreover, CM2 and Trans-M2 showed better mineralization-accelerating and rescuing effects when compared to their controls, and application of p38 inhibitor partially blocked the promotion.Our study demonstrated the inflammation-targeting and mineralization-promoting effects of M2 macrophages on cementoblasts, which may provide evidence for M2 macrophage-based cementum regeneration.

Introduction: GARP (Glycoprotein-A repetitions predominant) is a cell surface receptor which is coded by Lrrc32 (Leucine rich repeat containing protein 32) and involved in the activation of TGF-beta. The expression of GARP has been widely found in various cells including Treg, activated B cells, mesenchymal stromal cells and many types of cancer cells. In the tumor microenvironment (TME), GARP from Treg promotes the growth of tumor via secretion of immune suppressive TGF-beta. Result: We established GARP humanized mouse model (hGARP) on C57BL/6 background in which the signal peptide and extracellular domain of mouse GARP (mGARP) was replaced by human GARP (hGARP). Similar expression level of hGARP on Treg cells in splenocytes from C57BL/6-hGARP mice was observed compared with mGARP in wildtype mice. After treating with anti-hGARP antibody, tumor growth was inhibited in the cohort engrafted with MC38 colorectal cancer cell line. Flow cytometry analysis of the tumor infiltrating lymphocytes revealed an abundant accumulation of hGARP on the Treg cells suggesting that blocking of hGARP in Treg cells contributed to the therapeutic efficiency. In addition, hGARP expression was also found on platelet cells from C57BL/6-hGARP mice, which is similar to human. After treating with anti-hGARP, we also found a significant reduction of platelet cells in C57BL/6-hGARP mice. In vivo assay suggested that the reduction of platelet cells from C57BL/6-hGARP mice correlated well with the binding of anti-hGARP. Conclusion: C57BL/6-hGARP is a well-established mouse model for the pre-clinical evaluation of both therapeutic effect and toxicity of anti-hGARP therapy. Citation Format: Xing Liu, Meirong Wu, Huiyi Wang, Weiwei Yu, Jianming Xu, Hongyan Sun, Cunxiang Ju, Hongyu Wang, Santi Suryani Chen, Zhiying Li, Mark Wade Moore, Jing Zhao, Xiang Gao. Humanized mice for preclinical efficacy evaluation of drugs targeting GARP. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5198.

Severe immunodeficient mice engrafted with human hematopoietic stem cells (HSC) have been extensively used in immuno-oncology related studies to evaluate the efficacy of cancer therapies. However, because of species differences, murine cytokines provide limited support to human immune cells, thus immunodeficient mice have limited engraftment of human immune cells. Increasing evidence has shown that myeloid cells, especially macrophages and dendritic cells, are critical for the induction of anti-tumor immunity. We developed a mouse model, NCG-M, that can support human T, B, NK and myeloid cells such that in vivo evaluation of agents that require the interplay between these immune cells can be examined. This model was genetically engineered on the severe immunodeficient strain NCG and can produce human granulocyte/macrophage colony stimulating factor 2 (GM-CSF, also known as CSF2), interleukin-3 (IL-3) and stem cell factor (SCF, also known as KITLG). Upon human CD34+ HSC cell engraftment, increased myeloid lineage cells, such as granulocytes, monocytes, neutrophils, macrophages, and dendritic cells, were observed in the NCG-M cohort compared to NCG mice. The NCG-M mouse also supports the development of human T cells, and preliminary data showed increased B and NK cells. The NCG-M is an appropriate mouse model for studying the efficacy of therapeutic agents that require human T cells and myeloid cells. Citation Format: Huiyi Wang, Jialu Fan, Shiying Guo, Mingkun Zhang, Shuai Li, Santi Suryani Chen, Zhiying Li, Mark Wade Moore, Jing Zhao, Xiang Gao, Cunxiang Ju. NCG-M humanized mice: an excellent model for human immune reconstitution of myeloid lineages. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5202.

Human immune cell reconstitution (PBMC or CD34+ hematopoietic stem cell; HSC) is commonly performed in immunodeficient mouse strains such as NCG, NSG, or NOG. The use of PBMC is preferred for multiple reasons, including faster engraftment, lower cost, and the presence of mature immune cell populations. Although there are donor-to-donor variations in GvHD occurrence, and specific donors can be screened, the symptoms of GvHD typically occur within 4 weeks. GvHD significantly limits the study window required to evaluate the efficacy of therapeutic agents, such as cancer immunotherapies. GvHD is an immune reaction triggered mainly by donor T cells identifying the mouse MHC class I and class II as foreign and attacking the host cells and organs. Deletion of β2m (NCG- β2m-KO) reduces the occurrence of GvHD; however, β2m is not only in the MHC class I subunit but also in the FcRn subunit, of which deletion shortens the half-life of IgG, making it unsuitable for IgG antibody agent evaluation. In addition, the deletion of MHC class I or class II alone results in an imbalanced CD4/CD8 ratio. To solve these problems, we developed the NCG-MHC-dKO mouse model by knocking out the H2K1, H2D1, and H2Ab1 genes. Compared with the existing NCG- β2m-KO or other β2m null mouse models, the NCG-MHC-dKO significantly prolonged survival, reduced GvHD occurrence, and did not affect the CD4/CD8 ratio when reconstituted with PBMC. Furthermore, treatment with anti-PD-L1 antibodies significantly inhibited MDA-MB-231 tumor cell growth in PBMC-reconstituted NCG-MHC-dKO mice, similar to NCG mice. Based on our preliminary data, the NCG-MHC-dKO mouse is a promising model for antibody and cell therapy agent evaluation, assessing treatment-related cytokine release syndrome, and evaluating long-term toxicities of cell therapies. Citation Format: Huiyi Wang, Jun Xing, Jialu Fan, Huanhuan Hou, Santi Suryani Chen, Zhiying Li, Mark Wade Moore, Jing Zhao, Xiang Gao, Cunxiang Ju. NCG-MHC-dKO mice: an excellent model for PBMC reconstitution and pharmacodynamic evaluation in the absence of GvHD. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5192.

Despite the success of programmed death ligand 1 (PD-L1) and programmed death 1 (PD-1) blockage as cancer therapies, the overall clinical response rates are still unsatisfying, with only a 25% response rate in most cancers. The underlying mechanisms of non-responsiveness towards anti-PD-L1 treatment have been extensively studied and associated with many factors, including PD1/PD-L1 expression level, tumor mutation burden, IFN signaling, loss of MHC-I, tumor microenvironment (immune infiltration, suppressive immune system), and so on. Immunodeficient mice such as the NSG and NCG are commonly used to study the efficacy of cancer immunotherapies. These mice, reconstituted with either human PBMC or HSC, act as an avatar to provide a window of the potential agent efficacy at the preclinical level. However, due to the challenge of obtaining an HSC/PBMC donor, and often cost and time can also be a limiting factor, many studies only use 2-5 human donors. Here we present data demonstrating the significant variation observed between donor-to-donor, highlighting the importance of testing multiple donors. We reconstituted immune system and tumor tissue humanized mouse models using peripheral blood mononuclear cells (PBMCs), and cell line-derived xenografts (CDX) in severe immunodeficiency NCG mice. Anti-tumor efficacy of anti-PD-L1 (Tecentriq) in huPBMC-NCG mice engrafted with several tumor cells including MDA-MB-231, HCC827, NUGC4, and some other CDX cell lines was tested in our system. In the cases of MDA-MB-231, HCC827, and NUGC4, different donors were enrolled in the same study, and tumor growth inhibition measured by tumor volume (TGITV) of 3% to 52.40% (MDA-MB-231), 5% to 33% (HCC827) and 4.75% to 20.74% (NUGC4) were observed. Especially, In the case of MDA-MB-231, 6 donors were enrolled in this study at the same time. About 40% of immune infiltration was first observed in tumors. High expression of PD1 on immune cells and PDL1 on tumor cells were also detected from different donors simultaneously. Although the similar immune infiltration and PD1/PDL1 expression, the TGITV of 6 donors varied between 3% and 52.40%, suggesting the donor-dependent response to anti-PD-L1 treatment, which is similar to the clinical trials. Our data showed diverse in vivo responses when multiple human PBMC donors were used for preclinical agent evaluation. While unsurprising, it showed the critical need to include various human PBMC donors when an agent is in preclinical status because the donors' immune profile may dictate the study's outcome. This can be divided into two parts, on the one hand, donor screening for favorable efficacy response donors in advance is an alternative approach to ensure the positive efficacy of anti-PDL1 blockade relative therapy. On the other hand, the low-efficacy response donors also can be candidates for anti-PD1/PDL1 combination therapy. Citation Format: Tingting Gu, Weiwei Yu, Hongyan Sun, Fang Zhu, Huiyi Wang, Mengting Wang, Shuai Li, Jianming Xu, Santi Chen, Zhiying Li, Mark Wade Moore, Cunxiang Ju, Hongyu Wang, Jing Zhao, Xiang Gao. Analysis of selective response of anti-PD-L1 treatment in huPBMC-NCG mice reconstituted with different donors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6653.

Tumor necrosis factor (TNF) receptor 2 (TNFR2) is mainly expressed on the surface of regulatory T cells (Tregs) and CD8+ T cells, which can stimulate the proliferation of these cells through NF-kB activation. TNFR2 is also highly expressed on the surface of various human tumors, such as colorectal cancer and lung cancer. Therefore, a TNFR2 antagonist blocking the binding of TNFR2 to its ligand that targets TNFR2+ tumor-infiltrating Tregs can directly enhance the killing ability of effector T cells (Teff) against TNFR2-expressing tumors. Many pharmaceutical and biotech companies have lined up TNFR2-targeting drugs, and most TNFR2 antibodies related research is in the early preclinical/clinical stage. We established a humanized TNFR2 (hTNFR2) genetically engineered mouse model on the BALB/c background, in which the extracellular domain of the mouse TNFR2 was replaced with its human counterpart, and the murine transmembrane and cytoplasmic domain kept intact. The hTNFR2 chimeric protein is mainly expressed on Teff and Treg cells in BALB/c-hTNFR2 and BALB/c-hPD1/hPDL1/hTNFR2 mice, which is similar to mouse TNFR2 protein expression profiles in wild-type mice. Our data showed that anti-TNFR2 antibodies could significantly inhibit tumor growth in BALB/c-hTNFR2 mice bearing CT26 tumor cells or BALB/c-hTNFR2 mice bearing CT26-hTNFR2 tumors. Furthermore, within the tumor-infiltrating lymphocytes, there was a reduction in Tregs cells and an increase in cytotoxic CD8 T cells, which accurately mimics the mechanism of action of anti-TNFR2 antibodies. In conclusion, BALB/c-hTNFR2 and BALB/c-hPD1/hPDL1/hTNFR2 mice are ideal models for studying the efficacy and pharmacodynamics of anti-TNFR2 antibodies as a single agent or in combination with anti-PD-1/PD-L1 therapies. Citation Format: Huiyi Wang, Ying Li, Mingkun Zhang, Xu Su, Santi Suryani Chen, Zhiying Li, Mark Wade Moore, Jing Zhao, Xiang Gao, Cunxiang Ju. TNFR2 humanized mice: an ideal model for studying anti-TNFR2 antibodies. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5201.

Introduction: IL-15 is a four α-helix bundle cytokine which is produced by dendritic cells, monocytes, and epithelial cells. IL-15 is necessary for the development, survival, and activation of natural killer (NK) cells. NK cells are unable to persist in immunodeficient NCG mice reconsituted with human NK cells, but after knocking in human IL-15 (hIL-15), NCG-hIL15 mice can support the reconstitution and survival of human NK cells. Results: Compared with NCG mice, the level of hIL-15 was significantly increased in NCG-hIL15 mice. After transplanting human PBMC (huPBMC) or NK (huNK) cells into NCG-hIL15 mice, the peripheral blood level of NK cells in huNK-NCG-hIL15 mice was much higher than in huPBMC-NCG-hIL15 mice. The expression of perforin in peripheral blood of huNK-NCG-hIL15 mice was significantly higher than that in huPBMC-NCG-hIL15 mice. Considering the obvious ADCC of rituximab acting on Raji cells in vitro, we evaluated the efficacy of rituximab in huNK-NCG-hIL15 mice subcutaneously engrafted with Raji cells. Rituximab significantly inhibited the tumor growth in huNK-NCG-hIL15 mice. Conclusion: NCG-hIL15 is a great mouse model to evaluate the anti-tumor efficacy of drugs targeting human NK cells without the interference of T cells. Citation Format: Xing Liu, Meirong Wu, Huiyi Wang, Weiwei Yu, Jianming Xu, Hongyan Sun, Cunxiang Ju, Hongyu Wang, Santi Suryani Chen, Zhiying Li, Mark Wade Moore, Jing Zhao, Xiang Gao. NCG-hIL15 humanized mice: an ideal model for human immune reconstitution of NK cells. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3970.

Functional mature proteins usually undergo folding, and in vivo folding is often affected by factors such as translation ratio, intracellular environment, and chaperones. d -psicose 3-epimerase (DPEase) derived from Dorea sp. has excellent properties in catalytic production of d -allulose; a rare sugar with unique biological functions. Here, the Dorea sp. DPEase coding gene was fusion-expressed with four native chaperones (DnaK, DnaJ, GrpE and PrsA) in Bacillus subtilis, and resulted in an 4.31- and 6.12-fold increase to 10.81 U/g and 13.34 U/g in recombinant cells of BS-DPE-DnaK and BS-DPE-PrsA, respectively. The recombinant proteins showed increased thermostability without significant change in the optimum pH and Co2+ affinity. Using recombinant cells as catalysts, a 1.12–2.31-fold increase was detected in DPEase activity after treatment with 40 % ethanol. We showed that fusion expression of molecular chaperones to enhance protein-assisted folding could facilitate expression of DPEase. We hope that our results provide a facilitated route for large-scale production of d -allulose in B. subtilis.

PIWI-interacting small RNAs (piRNAs) protect the germline genome and are essential for fertility. piRNAs originate from transposable element (TE) RNAs, long non-coding RNAs, or 3´ untranslated regions (3´UTRs) of protein-coding messenger genes, with the last being the least characterized of the three piRNA classes. Here, we demonstrate that the precursors of 3´UTR piRNAs are full-length mRNAs and that post-termination 80S ribosomes guide piRNA production on 3´UTRs in mice and chickens. At the pachytene stage, when other co-translational RNA surveillance pathways are sequestered, piRNA biogenesis degrades mRNAs right after pioneer rounds of translation and fine-tunes protein production from mRNAs. Although 3´UTR piRNA precursor mRNAs code for distinct proteins in mice and chickens, they all harbor embedded TEs and produce piRNAs that cleave TEs. Altogether, we discover a function of the piRNA pathway in fine-tuning protein production and reveal a conserved piRNA biogenesis mechanism that recognizes translating RNAs in amniotes., Ribosome can mediate piRNA biogenesis from long non-coding RNAs following translation of the short open reading frames. Here the authors show that 80S ribosome also guides piRNA production from 3’ UTR of protein-coding genes after translation of long open reading frames, indicating a general piRNA biogenesis mechanism regardless of their precursor ORF length.

d-allulose, a rare sugar that is scarce in nature, exerts several beneficial effects and has commercial potential. d-allulose 3-epimerase (DAEase) plays a vital role in catalyzing the isomerization from d-fructose to d-allulose. However, the industrial application of DAEase for d-allulose production is hindered by its poor long-term thermostability. In the present research, we introduced a proline residue (i) to restrict its spatial conformation and (ii) to reduce the entropy of the unfolded state of DAEase. The t

This paper presents a reconfigurable leaky wave antenna (LWA) that can alter the main beam angle at a fixed frequency. To further realize the miniaturization of the LWA, a half-mode bent corrugated substrate integrated waveguide (HM-BCSIW) is applied in this design. Equivalent circuit parameters of the antenna are adjusted by loading varactor diodes, which enable beam scanning at a fixed frequency of 5.7GHz. The radiation angle of the design varies from +55° to −27° continuously, along with a biasing voltage from 2 V to 18 V.

Skin penetration of active molecules for treatment of diverse diseases is a major field of research owing to the advantages associated with the skin like easy accessibility, reduced systemic-derived side effects, and increased therapeutic efficacy. Despite these advantages, dermal drug delivery is generally challenging due to the low skin permeability of therapeutics. Although various methods have been developed to improve skin penetration and permeation of therapeutics, they are usually aggressive and could lead to irreversible damage to the stratum corneum. Nanosized carrier systems represent an alternative approach for current technologies, with minimal damage to the natural barrier function of skin. In this Review, the use of nanoparticles to deliver drug molecules, genetic material, and vaccines into the skin is discussed. In addition, nanotoxicology studies and the recent clinical development of nanoparticles are highlighted to shed light on their potential to undergo market translation. We are grateful to Maria Angela Motta and Oliver Etzold for their valuable scientific inputs and corrections in the review. We are thankful to MINECO (RTI2018-098951-B-I00), IKERBASQUE-Basque Foundation for Science, Basque Government (Elkartek projects KK-2019/00086 and KK2020/00010; proyectos de investigacion basica/aplicada PIBA_2020_1_0056), Gipuzkoa Provincial Council (2019CIEN-000075-01), and to Polymat, Basque Centre for Macromolecular Design and Engineering for funding. A.S. acknowledges financial support from CONICET, ANPCyT, Universidad Nacional del Litoral. H.W. is thankful for the financial support received from China Scholarship Council (File no. 201804910606).

Hematopoietic stem cell (HSC) transplantation is an adoptive immunotherapy for treating malignant hematopoietic cancers and autoimmune disorders. Preclinical studies of such novel therapies require animal models that enable and sustain human erythrocyte reconstitution. Existing mouse models typically require irradiation to remove mouse HSCs from the bone marrow prior to human immune system reconstitution. Irradiation adversely affects the health of the animal by promoting necrosis and apoptosis of gastrointestinal, neural and muscle tissues, which can lead to wasting, infection, and even death. An irradiation-free mouse model for human erythroid reconstitution is in urgent need. KIT proto-oncogene receptor tyrosine kinase, also known as c-KIT, is an important cell surface marker expressed in both mouse and human HSCs, multipotent progenitors (MPP), and common myeloid progenitors (CMP). KIT signaling plays a key role in cell survival, proliferation, and differentiation. Alterned forms of this protein have been shown to be associated with some types of cancer. Point mutation of Val831Met is known to cause macrocytic anemia. We speculate that mice with this mutation experience spontaneous anemia and inhibited HSC function, which would support human erythroid reconstitution without irradiation. NCG-X mouse was generated based on this hypothesis, building upon the severe immunodeficient NCG generated by GemPharmatech years ago. This new strain of mice are deficient of T/B/NK cells with functional inhibition of HSCs. Here, we report that NCG-X supports human HSC transplantation without radiation, enabling complete avoidance of adverse effects on animal health. When compared with irradiated NCG mice, NCG-X mice are able to not only rebuild the human immune system, but also enable human erythroid cells to reconstruct the bone marrow for a prolonged period of time after engraftment: 16 and 30 weeks post-engraftment have been assayed. Though reduced red blood cells and hemoglobin have been observed, 30 weeks post-engraftment enables consistent support from the start to the finish of the preclinical phase. In conclusion, NCG-X provides an improved and preclinical relevant mouse model for therapeutic development of leukemia, thalassemia, other types of blood diseases and autoimmune diseases. Citation Format: Tingting Gu, Cunxiang Ju, Hongyan Sun, Xiang Gao, MingKun Zhang, Weiwei Yu, Mengting Wang, Shuai Li, Shuai Li, Huiyi Wang, Jing Zhao. NCG-X mouse: A novel animal model to evaluate preclinical studies of humanized erythroid reconstitution without irradiation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5621.

GITR (glucocorticoid-induced TNFR-related protein) is a member of tumor necrosis factor receptor superfamily, expressed on many kinds of cells including T cells, attracting attention as a co-stimulatory immune checkpoint molecule. GITRL, the ligand of GITR is mainly expressed on antigen presenting cells. GITR/GITRL interaction plays an important role on the development of immune response, which can serve as promising anti-tumor target. While more than twenty anti-GITR antibodies are at clinical stages, combination treatments of GITR agonist and PD-1 inhibitor may be a better choice. Thus, there is a need for pre-clinical mouse model to evaluate the anti-tumor effect of humanized PD1/GITR antibodies. Gempharmatech established PD1/GITR double humanized mouse line on a BALB/c background, in which the extracellular domain of mGITR was replaced with its human counterparts while the trans-membrane and cytoplasmic domain kept intact, as well as the same strategy of PD1. BALB/c-hPD1/hGITR mouse could successfully express similar levels of hGITR and hPD1 compared with BALB/c wild type mice. Moreover, no immune or developmental abnormalities were observed in the humanized mouse. Anti-GITR combination with anti-PD1 presented greater tumor growth inhibition than anti-PD1 or anti-GITR monotherapies in PD1/GITR double humanized mouse bearing CT26 tumors. In summary, BALB/c-hPD1/hGITR are valuable models for the pre-cilincal evaluation of candidate humanized PD1/GITR combination treatment or bispecific antibody therapies. Citation Format: Dongdong Yuan, Dongdong zhang, Hongyan Sun, Mingkun Zhang, Liyou Dong, Cunxiang Ju, Shuai Li, Huiyi Wang, Jing Zhao, Xiang Gao. Preclinical evaluation of humanized PD1/GITR combination therapy using double humanized mouse [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5622.

Synthesis, characterization, and coating performance of castor oil (CO)-based waterborne hyperbranched polyurethane acrylate (WHPUA) emulsions are highlighted in this work. Herein, a second-generation CO-based hyperbranched polyester (C20) was synthesized by introducing the renewable CO as a B3 core and dimethylolpropionic acid as an AB2 monomer via a pseudo-one-pot condensation procedure. Three types of CO-based WHPUA emulsions were prepared through the addition of the hyperbranched C20 with maleic anhydride (MA) and the isocyanate-bearing semiadduct (IPDI-HEA) at different MA/IPDI-IH molar ratios without the addition of any emulsifiers. The prepared emulsions were further formulated into UV-curable coating formulations and exposed to a medium-pressure mercury lamp. Consequently, the obtained UV-cured coatings exhibited outstanding adhesion performance, excellent transparency, and an acceptable degree of final double bond conversion. Owing to a relatively low concentration of hydrophilic units and a large number of acrylate functionalities in the waterborne oligomer, the obtained films were endowed with high crosslinking density and low hydrophilic nature, thus producing enhanced tensile strength, pencil hardness, water resistance, and chemical resistance. Due to their combination of renewable materials, a waterborne system and a UV-curing technique, these CO-based WHPUA emulsions offer promising opportunities to be used as environmentally friendly coatings.

To help curb the spread of the COVID-19 pandemic, governments and public health authorities around the world have launched a number of contact-tracing apps. Although contact tracing apps have received extensive attentions from the research community, no existing work has characterized the users' adoption of contact tracing apps from the app market level. In this work, we perform the first market-level analysis of contact tracing apps. We perform a longitudinal empirical study (over 4 months) of eight government-backed COVID-19 contact tracing apps in iOS app store. We first collect all the daily meta information (e.g., app updates, app rating, app comments, etc.) of these contact tracing apps from their launch to 2020-07-31. Then we characterize them from release practice, app popularity, and mobile users' feedback. Our study reveals various issues related to contact tracing apps from the users' perspective, hoping to help improve the quality of contact tracing apps and thus achieving a high level of adoption in the population.

The keystone pathogen Porphyromonas gingivalis (P. gingivalis) elicits inflammation and autophagy in periodontal tissues. Transcription factor CXXC-type zinc finger protein 5 (CXXC5) and various signals are sensitive to P. gingivalis invasion. Herein, we investigated the P. gingivalis-elicited autophagy activity, the contribution of CXXC5, and the involvement of signals in cementoblasts, tooth root surface cells crucial in periodontal and periapical regions. After coculture with P. gingivalis, cementoblasts exhibited inflammatory cytokine increase, light chain 3(LC3)-I/II conversion, autophagosome activation, and CXXC5 reduction. Cementoblasts with loss and gain of CXXC5 were developed. CXXC5 silencing suppressed autophagy and inflammation, thereby partially compensating for the effects of P. gingivalis, and vice versa. We then screened potential signals and verified the positive participation of Stat3/Akt/Erk networks through specific inhibitor employment. P. gingivalis and CXXC5 induced autophagy through Beclin1 and Atg5 activation. Intriguingly, Annexin V/PI assay and EdU detection revealed that P. gingivalis promoted apoptosis and repressed cell proliferation. In sum, coculture with P. gingivalis enhanced autophagy activity in cementoblasts, which was partially suppressed by CXXC5 downregulation and mediated by Jak/Stat3, PI3K-Akt, and Erk1/2 signaling. This process probably influenced cell apoptosis and proliferation.