Splice variant of growth hormone-releasing hormone receptor drives esophageal squamous cell carcinoma conferring a therapeutic target.

The extrahypothalamic growth hormone-releasing hormone (GHRH) and its cognate receptors (GHRH-Rs) and splice variants are expressed in a variety of cancers. It has been shown that the pituitary type of GHRH-R (pGHRH-R) mediates the inhibition of tumor growth induced by GHRH-R antagonists. However, GHRH-R antagonists can also suppress some cancers that do not express pGHRH-R, yet the underlying mechanisms have not been determined. Here, using human esophageal squamous cell carcinoma (ESCC) as a model, we were able to reveal that SV1, a known splice variant of GHRH-R, is responsible for the inhibition induced by GHRH-R antagonist MIA-602. We demonstrated that GHRH-R splice variant 1 (SV1) is a hypoxia-driven promoter of tumor progression. Hypoxia-elevated SV1 activates a key glycolytic enzyme, muscle-type phosphofructokinase (PFKM), through the nuclear factor kappa B (NF-κB) pathway, which enhances glycolytic metabolism and promotes progression of ESCC. The malignant actions induced by the SV1-NF-κB-PFKM pathway could be reversed by MIA-602. Altogether, our studies demonstrate a mechanism by which GHRH-R antagonists target SV1. Our findings suggest that SV1 is a hypoxia-induced oncogenic promoter which can be an alternative target of GHRH-R antagonists.

Modification of pore-wall in direct ink writing wollastonite scaffolds favorable for tuning biodegradation and mechanical stability and enhancing osteogenic capability.

Surface chemistry and mechanical stability determine the osteogenic capability of bone implants. The development of high-strength bioactive scaffolds for in-situ repair of large bone defects is challenging because of the lack of satisfying biomaterials. In this study, highly bioactive Ca-silicate (CSi) bioceramic scaffolds were fabricated by additive manufacturing and then modified for pore-wall reinforcement. Pure CSi scaffolds were fabricated using a direct ink writing technique, and the pore-wall was modified with 0%, 6%, or 10% Mg-doped CSi slurry (CSi, CSi-Mg6, or CSi-Mg10) through electrostatic interaction. Modified CSi@CSi-Mg6 and CSi@CSi-Mg10 scaffolds with over 60% porosity demonstrated an appreciable compressive strength beyond 20 MPa, which was ~2-fold higher than that of pure CSi scaffolds. CSi-Mg6 and CSi-Mg10 coating layers were specifically favorable for retarding bio-dissolution and mechanical decay of scaffolds in vitro. In-vivo investigation of critical-size femoral bone defects repair revealed that CSi@CSi-Mg6 and CSi@CSi-Mg10 scaffolds displayed limited biodegradation, accelerated new bone ingrowth (4-12 weeks), and elicited a suitable mechanical response. In contrast, CSi scaffolds exhibited fast biodegradation and retarded new bone regeneration after 8 weeks. Thus, tailoring of the chemical composition of pore-wall struts of CSi scaffolds is beneficial for enhancing the biomechanical properties and bone repair efficacy.

Concurrent arthroscopic meniscal repair during open-wedge high tibial osteotomy is not clinically beneficial for medial meniscus posterior root tears.

PURPOSE: This prospective study aimed to investigate the clinical benefits of meniscal repair during open-wedge high tibial osteotomies (OWHTOs) in patients with medial meniscus posterior root tears (MMPRTs) and to identify potential risk factors for meniscal healing. METHODS: Ninety patients with degenerative MMPRTs were included in the final cohort and randomized into three groups. The patients in Group A (n = 30) underwent OWHTO and arthroscopic all-inside meniscal repair concurrently, those in Group B (n = 34) underwent OWHTO only, and those in Group C (n = 26) underwent arthroscopic partial meniscectomy. Clinical and radiological outcomes were recorded, and meniscal healing was evaluated during second-look arthroscopy. Logistic regression analysis was performed to identify risk factors for meniscal healing. RESULTS: After a minimum follow-up of 24 months, no significant differences between Groups A and B regarding the final Lysholm (p = 0.689) or Hospital for Special Surgery (HSS) scores (p = 0.256) were observed. There were significant differences among the three groups regarding the hip-knee-ankle angle (HKA), weight-bearing line (WBL) ratio, medial proximal tibial angle (MPTA), and joint line convergence angle (JLCA) (p < 0.001, respectively), but the differences between Groups A and B were not significant. During second-look arthroscopy, the healing rate of the MMPRTs was significantly higher in Group A (63.3%) than in Group B (35.3%). Concurrent meniscal repair and changes in the HKA, and MPTA were risk factors for meniscal healing. CONCLUSION: Concurrent arthroscopic meniscal repair during OWHTO did not lead to significant clinical benefits in the treatment of MMPRTs, except for an increased rate of meniscal healing, which was not associated with clinical outcomes. LEVEL OF EVIDENCE: II, prospective comparative study.

Growth hormone-releasing hormone receptor antagonists inhibit human gastric cancer through downregulation of PAK1-STAT3/NF-κB signaling.

Gastric cancer (GC) ranks as the fourth most frequent in incidence and second in mortality among all cancers worldwide. The development of effective treatment approaches is an urgent requirement. Growth hormone-releasing hormone (GHRH) and GHRH receptor (GHRH-R) have been found to be present in a variety of tumoral tissues and cell lines. Therefore the inhibition of GHRH-R was proposed as a promising approach for the treatment of these cancers. However, little is known about GHRH-R and the relevant therapy in human GC. By survival analyses of multiple cohorts of GC patients, we identified that increased GHRH-R in tumor specimens correlates with poor survival and is an independent predictor of patient prognosis. We next showed that MIA-602, a highly potent GHRH-R antagonist, effectively inhibited GC growth in cultured cells. Further, this inhibitory effect was verified in multiple models of human GC cell lines xenografted into nude mice. Mechanistically, GHRH-R antagonists target GHRH-R and down-regulate the p21-activated kinase 1 (PAK1)-mediated signal transducer and activator of transcription 3 (STAT3)/nuclear factor-κB (NF-κB) inflammatory pathway. Overall, our studies establish GHRH-R as a potential molecular target in human GC and suggest treatment with GHRH-R antagonist as a promising therapeutic intervention for this cancer.

Dysregulation of PAK1 Is Associated with DNA Damage and Is of Prognostic Importance in Primary Esophageal Small Cell Carcinoma.

Primary esophageal small cell carcinoma (PESCC) is a rare, but fatal subtype of esophageal carcinoma. No effective therapeutic regimen for it. P21-activated kinase 1 (PAK1) is known to function as an integrator and an indispensable node of major growth factor signaling and the molecular therapy targeting PAK1 has been clinical in pipeline. We thus set to examine the expression and clinical impact of PAK1 in PESCC. The expression of PAK1 was detected in a semi-quantitative manner by performing immunohistochemistry. PAK1 was overexpressed in 22 of 34 PESCC tumors, but in only 2 of 18 adjacent non-cancerous tissues. Overexpression of PAK1 was significantly associated with tumor location (p = 0.011), lymph node metastasis (p = 0.026) and patient survival (p = 0.032). We also investigated the association of PAK1 with DNA damage, a driven cause for malignancy progression. γH2AX, a DNA damage marker, was detectable in 18 of 24 (75.0%) cases, and PAK1 expression was associated with γH2AX (p = 0.027). Together, PAK1 is important in metastasis and progression of PESCC. The contribution of PAK1 to clinical outcomes may be involved in its regulating DNA damage pathway. Further studies are worth determining the potentials of PAK1 as prognostic indicator and therapeutic target for PESCC.

New insight into biodegradable macropore filler on tuning mechanical properties and bone tissue ingrowth in sparingly dissolvable bioceramic scaffolds.

Structural parameters of the implants such as shape, size, and porosity of the pores have been extensively investigated to promote bone tissue repair, however, it is unknown how the pore interconnectivity affects the bone growth behaviors in the scaffolds. Herein we systematically evaluated the effect of biodegradable bioceramics as a secondary phase filler in the macroporous networks on the mechanical and osteogenic behaviors in sparingly dissolvable bioceramic scaffolds. The pure hardystonite (HT) scaffolds with ∼550 & 800 µm in pore sizes were prepared by digital light processing, and then the Sr-doped calcium silicate (SrCSi) bioceramic slurry without and with 30 % organic porogens were intruded into the HT scaffolds with 800 µm pore size and sintered at 1150 °C. It indicated that the organic porogens could endow spherical micropores in the SrCSi filler, and the invasion of the SrCSi component could not only significantly enhance the compressive strength and modulus of the HT-based scaffolds, but also induce osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). The pure HT scaffolds showed extremely slow bio-dissolution in Tris buffer after immersion for 8 weeks (∼1 % mass decay); in contrast, the SrCSi filler would readily dissolve into the aqueous medium and produced a steady mass decay (>6 % mass loss). In vivo experiments in rabbit femoral bone defect models showed that the pure HT scaffolds showed bone tissue ingrowth but the bone growth was impeded in the SrCSi-intruded scaffolds within 4 weeks; however, the group with higher porosity of SrCSi filler showed appreciable osteogenesis after 8 weeks of implantation and the whole scaffold was uniformly covered by new bone tissues after 16 weeks. These findings provide some new insights that the pore interconnectivity is not inevitable to impede bone ingrowth with the prolongation of implantation time, and such a highly biodegradable and bioactive filler intrusion strategy may be beneficial for optimizing the performances of scaffolds in bone regenerative medicine applications.

Application of 3D-bioprinted nanocellulose and cellulose derivative-based bio-inks in bone and cartilage tissue engineering.

212Three-dimensional (3D) printing is a modern, computer-aided, design-based technology that allows the layer-by-layer deposition of 3D structures. Bioprinting, a 3D printing technology, has attracted increasing attention because of its capacity to produce scaffolds for living cells with extreme precision. Along with the rapid development of 3D bioprinting technology, the innovation of bio-inks, which is recognized as the most challenging aspect of this technology, has demonstrated tremendous promise for tissue engineering and regenerative medicine. Cellulose is the most abundant polymer in nature. Various forms of cellulose, nanocellulose, and cellulose derivatives, including cellulose ethers and cellulose esters, are common bioprintable materials used to develop bio-inks in recent years, owing to their biocompatibility, biodegradability, low cost, and printability. Although various cellulose-based bio-inks have been investigated, the potential applications of nanocellulose and cellulose derivative-based bio-inks have not been fully explored. This review focuses on the physicochemical properties of nanocellulose and cellulose derivatives as well as the recent advances in bio-ink design for 3D bioprinting of bone and cartilage. In addition, the current advantages and disadvantages of these bio-inks and their prospects in 3D printing-based tissue engineering are comprehensively discussed. We hope to offer helpful information for the logical design of innovative cellulose-based materials for use in this sector in the future.

The complete plastome sequence of Artocarpus altilis (Parkinson ex F.A. Zorn) Fosberg, (Moraceae).

Artocarpus altilis (Parkinson ex F.A. Zorn) Fosberg is native to the Pacific Islands, India, and the Philippines. It is also cultivated in Taiwan and Hainan. The complete plastome of the species was assembled and annotated in this study. The circular genome was 160,184 bp in size, presenting a typical quadripartite structure including two inverted repeats (IRs) of 25,734 bp, a large single-copy (LSC) of 88,791 bp, and a small single-copy (SSC) of 19,925 bp. The genome contained 132 genes, including 87 protein-coding genes, 37 tRNA genes, and eight rRNA genes. The total G/C content of complete plastome was 36.0%, with the corresponding values of the LSC, SSC, and IR being 33.7%, 28.8%, and 42.7%, respectively. The complete plastome sequence of A. altilis (Parkinson ex F.A. Zorn) Fosberg will make contributions to the conservation genetics of this species as well as to phylogenetic studies of Moraceae.

A Novel Bispecific Antibody for EpCAM-Directed Inhibition of the CD73/Adenosine Immune Checkpoint in Ovarian Cancer.

PD-1/PD-L1-inhibiting antibodies have shown disappointing efficacy in patients with refractory ovarian cancer (OC). Apparently, OC cells exploit nonoverlapping immunosuppressive mechanisms to evade the immune system. In this respect, the CD73-adenosine inhibitory immune checkpoint is of particular interest, as it rapidly converts pro-inflammatory ATP released from cancer cells to immunosuppressive adenosine (ADO). Moreover, cancer-cell-produced ADO is known to form a highly immunosuppressive extra-tumoral 'halo' that chronically inhibits the anticancer activity of various immune effector cells. Thus far, conventional CD73-blocking antibodies such as oleclumab show limited clinical efficacy, probably due to the fact that it indiscriminately binds to and blocks CD73 on a massive surplus of normal cells. To address this issue, we constructed a novel bispecific antibody (bsAb) CD73xEpCAM that inhibits CD73 expressed on the OC cell surface in an EpCAM-directed manner. Importantly, bsAb CD73xEpCAM showed potent capacity to inhibit the CD73 enzyme activity in an EpCAM-directed manner and restore the cytotoxic activity of ADO-suppressed anticancer T cells. Additionally, treatment with bsAb CD73xEpCAM potently inhibited the proliferative capacity of OC cells and enhanced their sensitivity to cisplatin, doxorubicin, 5FU, and ionizing radiation. BsAb CD73xEpCAM may be useful in the development of tumor-directed immunotherapeutic approaches to overcome the CD73-mediated immunosuppression in patients with refractory OC.

Novel Fab-peptide-HLA-I fusion proteins for redirecting pre-existing anti-CMV T cell immunity to selective eliminate carcinoma cells.

Typically, anticancer CD8pos T cells occur at low frequencies and become increasingly impaired in the tumor micro environment. In contrast, antiviral CD8pos T cells display a much higher polyclonality, frequency, and functionality. In particular, cytomegalovirus (CMV) infection induces high numbers of 'inflationary' CD8pos T cells that remain lifelong abundantly present in CMV-seropositive subjects. Importantly, these so-called inflationary anti-CMV T cells increase with age, maintain a ready-to-go state, populate tumors, and do not become exhausted or senescent. Given these favorable attributes, we devised a novel series of recombinant Fab-peptide-HLA-I fusion proteins and coined them 'ReTARGs'. A ReTARG fusion protein consists of a high-affinity Fab antibody fragment directed to carcinoma-associated cell surface antigen EpCAM (or EGFR), fused in tandem with soluble HLA-I molecule/ß2-microglobulin, genetically equipped with an immunodominant peptide derived from CMV proteins pp65 (or IE-1). Decoration with EpCAM-ReTARGpp65 rendered EpCAM-expressing primary patient-derived carcinoma cells highly sensitive to selective elimination by cognate anti-CMV CD8pos T cells. Importantly, this treatment did not induce excessive levels of proinflammatory T cell-secreted IFNγ. In contrast, analogous treatment with equimolar amounts of EpCAM/CD3-directed bispecific T-cell engager solitomab resulted in a massive release of IFNγ, a feature commonly associated with adverse cytokine-release syndrome. Combinatorial treatment with EpCAM-ReTARGpp65 and EGFR-ReTARGIE-1 strongly potentiated selective cancer cell elimination owing to the concerted action of the corresponding cognate anti-CMV CD8pos T cell clones. In conclusion, ReTARG fusion proteins may be useful as an alternative or complementary form of targeted cancer immunotherapy for 'cold' solid cancers.

A triphasic biomimetic BMSC-loaded scaffold for osteochondral integrated regeneration in rabbits and pigs.

Osteochondral tissue involves cartilage, calcified cartilage and subchondral bone. These tissues differ significantly in chemical compositions, structures, mechanical properties and cellular compositions. Therefore, the repairing materials face different osteochondral tissue regeneration needs and rates. In this study, we fabricated an osteochondral tissue-inspired triphasic material, which was composed of a poly(lactide-co-glycolide) (PLGA) scaffold loaded with fibrin hydrogel, bone marrow stromal cells (BMSCs) and transforming growth factor-ß1 (TGF-ß1) for cartilage tissue, a bilayer poly(L-lactide-co-caprolactone) (PLCL)-fibrous membrane loaded with chondroitin sulfate and bioactive glass, respectively, for calcified cartilage, and a 3D-printed calcium silicate ceramic scaffold for subchondral bone. The triphasic scaffold was press-fitted into the osteochondral defects in rabbit (cylindrical defects with a diameter of 4 mm and a depth of 4 mm) and minipig knee joints (cylindrical defects with a diameter of 10 mm and a depth of 6 mm). The µ-CT and histological analysis showed that the triphasic scaffold was partly degraded, and significantly promoted the regeneration of hyaline cartilage after they were implanted in vivo. The superficial cartilage showed good recovery and uniformity. The calcified cartilage layer (CCL) fibrous membrane was in favor of a better cartilage regeneration morphology, a continuous cartilage structure and less fibrocartilage tissue formation. The bone tissue grew into the material, while the CCL membrane limited bone overgrowth. The newly generated osteochondral tissues were well integrated with the surrounding tissues too.

Comparison of osteogenic capability of 3D-printed bioceramic scaffolds and granules with different porosities for clinical translation.

Pore parameters, structural stability, and filler morphology of artificial implants are key factors influencing the process of bone tissue repair. However, the extent to which each of these factors contributes to bone formation in the preparation of porous bioceramics is currently unclear, with the two often being coupled. Herein, we prepared magnesium-doped wollastonite (Mg-CSi) scaffolds with 57% and 70% porosity (57-S and 70-S) via a 3D printing technique. Meanwhile, the bioceramic granules (57-G and 70-G) with curved pore topography (IWP) were prepared by physically disrupting the 57-S and 70-S scaffolds, respectively, and compared for in vivo osteogenesis at 4, 10, and 16 weeks. The pore parameters and the mechanical and biodegradable properties of different porous bioceramics were characterized systematically. The four groups of porous scaffolds and granules were then implanted into a rabbit femoral defect model to evaluate the osteogenic behavior in vivo. 2D/3D reconstruction and histological analysis showed that significant bone tissue production was visible in the central zone of porous granule groups at the early stage but bone tissue ingrowth was slower in the porous scaffold groups. The bone tissue regeneration and reconstruction capacity were stronger after 10 weeks, and the porous architecture of the 57-S scaffold was maintained stably at 16 weeks. These experimental results demonstrated that the structure-collapsed porous bioceramic is favorable for early-stage osteoconduction and that the 3D topological scaffolds may provide more structural stability for bone tissue growth for a long-term stage. These findings provide new ideas for the selection of different types of porous bioceramics for clinical bone repair.

Bispecific antibody CD73xEGFR more selectively inhibits the CD73/adenosine immune checkpoint on cancer cells and concurrently counteracts pro-oncogenic activities of CD73 and EGFR.

BACKGROUND: CD73 is an ecto-enzyme that is involved in the conversion of pro-inflammatory extracellular ATP (eATP) excreted by cancer cells under stress to anti-inflammatory adenosine (ADO). A broad variety of solid cancer types was shown to exploit CD73 overexpression as a suppressive immune checkpoint. Consequently, CD73-antagonistic antibodies, most notably oleclumab, are currently evaluated in several multicenter trials for clinical applicability. However, the efficacy of conventional monospecific CD73-inhibiting antibodies may be limited due to on-target/off-tumor binding to CD73 on normal cells. Therefore, a novel approach that more selectively directs CD73 immune checkpoint inhibition towards cancer cells is warranted. METHODS: To address this issue, we constructed a novel tetravalent bispecific antibody (bsAb), designated bsAb CD73xEGFR. Subsequently, the anticancer activities of bsAb CD73xEGFR were evaluated using in vitro and in vivo tumor models. RESULTS: In vitro treatment of various carcinoma cell types with bsAb CD73xEGFR potently inhibited the enzyme activity of CD73 (~71%) in an EGFR-directed manner. In this process, bsAb CD73xEGFR induced rapid internalization of antigen/antibody complexes, which resulted in a prolonged concurrent displacement of both CD73 and EGFR from the cancer cell surface. In addition, bsAb CD73xEGFR sensitized cancer to the cytotoxic activity of various chemotherapeutic agents and potently inhibited the proliferative/migratory capacity (~40%) of cancer cells. Unexpectedly, we uncovered that treatment of carcinoma cells with oleclumab appeared to enhance several pro-oncogenic features, including upregulation and phosphorylation of EGFR, tumor cell proliferation (~20%), and resistance towards cytotoxic agents and ionizing radiation (~39%). Importantly, in a tumor model using immunocompetent BALB/c mice inoculated with syngeneic CD73pos/EGFRpos CT26 cancer cells, treatment with bsAb CD73xEGFR outperformed oleclumab (65% vs 31% tumor volume reduction). Compared with oleclumab, treatment with bsAb CD73xEGFR enhanced the intratumoral presence of CD8pos T cells and M1 macrophages. CONCLUSIONS: BsAb CD73xEGFR outperforms oleclumab as it inhibits the CD73/ADO immune checkpoint in an EGFR-directed manner and concurrently counteracts several oncogenic activities of EGFR and CD73. Therefore, bsAb CD73xEGFR may be of significant clinical potential for various forms of difficult-to-treat solid cancer types.

The complete plastome of Garcinia subelliptica, Merr. 1909 (Clusiaceae).

The complete plastome of G. subelliptica, Merr. 1909. The complete length is 158,356 bp, with the typical structure and gene content of angiosperm plastomes, including a large single-copy region (LSC) of 86,220 bp, a repeat region (IRB), and a reverse repeat region (IRA) of 27,399 bp, respectively, and a small single-copy region (SSC) of 17,338 bp. The plastome contains 130 genes, including 85 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The total G/C content of the plastome is 36.1%.

Complete plastome sequence of Reevesia pycnantha Y. Ling (Malvaceae): a rare medicinal tree species in South Asia.

Here, we report and characterize the complete plastome of Reevesia pycnantha, Y. Ling 1951, which is a rare tree in the plant family Malvaceae. It is distributed in central and southern Jiangxi, Fujian, Guangdong, Hunan, and other provinces of China, where it is endemic. It grows in subtropical climates at middle and low altitudes of 200-800 meters within valleys, along mountain foothills, or on hillsides, in evergreen broad-leaved forests or at forest edges. Our results show that the length of the complete plastome is 161,964 bp, including 129 genes consisted of 81 protein-coding genes, 37 tRNA genes and 8 rRNA genes. It exhibits the typical quadripartite structure and gene content of angiosperms plastomes and comprises two inverted repeat (IRS) regions of 2,469 bp, a large single copy (LSC) region of 90,657 bp, and a small single-copy (SSC) region of 20,315 bp. The total G/C content in the plastome of R. pycnantha,Y. Ling 1951 is 36.8%. The complete plastome sequence of R. pycnantha, Y. Ling 1951will make contributions to the conservation genetics of this species as well as to phylogenetic studies in Malvaceae.

Complete plastome sequence from a cultivar of Diospyros nigra (J.F.Gmel.) M.R. Almeida (Ebenaceae): a nutritious fruit tree with high economic value cultivated in Hainan province, China.

Diospyros nigra (J.F.Gmel.) M.R.Almeida is a rare tree in the family Ebenaceae. The species is native to South America, while having been introduced to Florida and Texas (USA), India, Java and Madagascar. Additionally, this species is distributed in Guangdong Province and the southwest portion of Hainan Province, China. Here, we report and characterize the complete plastome of a cultivar of D. nigra. The length of the complete plastome is 157,168 bp, including 131 genes consisting of 84 protein-coding genes, 37 tRNA genes and 8 rRNA genes. The plastome has the typical structure and gene content of angiosperms, including two inverted repeat (IR) regions of 26,095 bp, a large single copy (LSC) region of 86,610 bp and a small single-copy (SSC) region of 18,386 bp. The total G/C content of the plastome in D. nigra is 37.4%. The complete plastome sequence of D. nigra will make contributions to the conservation genetics of the species, as well as to phylogenetic studies in Ebenaceae.

The complete chloroplast genome sequence of a Citrus australasica cultivar (Rutaceae）.

Citrus australasica (F. Muell.) Swingle belongs to the family Rutaceae. Citrus australasica is native to eastern Australia and southeastern New Guinea, and is mainly concentrated in a small region of northern New South Wales and tropical rainforest areas in southern Queensland. The complete plastome length of C. australasica is 160,335 bp, with the typical structure and gene content of angiosperm plastids, including a 26,592 bp repeat B (IRB) region, 26,952 bp IRA, 87,678 bp large single copy (LSC) region and 18,756 bp small single copy (SSC) region. The plastid contains 135 genes, including 89 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The total G/C content of the C. australasica plastome is 38.4%. The complete plastome sequence of C. australasica will provide useful resources for conservation genetics research of this species and phylogenetic research of Rutaceae.

Integrating pore architectures to evaluate vascularization efficacy in silicate-based bioceramic scaffolds.

Pore architecture in bioceramic scaffolds plays an important role in facilitating vascularization efficiency during bone repair or orbital reconstruction. Many investigations have explored this relationship but lack integrating pore architectural features in a scaffold, hindering optimization of architectural parameters (geometry, size and curvature) to improve vascularization and consequently clinical outcomes. To address this challenge, we have developed an integrating design strategy to fabricate different pore architectures (cube, gyroid and hexagon) with different pore dimensions (∼350, 500 and 650 µm) in the silicate-based bioceramic scaffolds via digital light processing technique. The sintered scaffolds maintained high-fidelity pore architectures similar to the printing model. The hexagon- and gyroid-pore scaffolds exhibited the highest and lowest compressive strength (from 15 to 55 MPa), respectively, but the cube-pore scaffolds showed appreciable elastic modulus. Moreover, the gyroid-pore architecture contributed on a faster ion dissolution and mass decay in vitro. It is interesting that both µCT and histological analyses indicate vascularization efficiency was challenged even in the 650-µm pore region of hexagon-pore scaffolds within 2 weeks in rabbit models, but the gyroid-pore constructs indicated appreciable blood vessel networks even in the 350-µm pore region at 2 weeks and high-density blood vessels were uniformly invaded in the 500- and 650-µm pore at 4 weeks. Angiogenesis was facilitated in the cube-pore scaffolds in comparison with the hexagon-pore ones within 4 weeks. These studies demonstrate that the continuous pore wall curvature feature in gyroid-pore architecture is an important implication for biodegradation, vascular cell migration and vessel ingrowth in porous bioceramic scaffolds.

Neoantigen-based cancer vaccination using chimeric RNA-loaded dendritic cell-derived extracellular vesicles.

Cancer vaccines critically rely on the availability of targetable immunogenic cancer-specific neoepitopes. However, mutation-based immunogenic neoantigens are rare or even non-existent in subgroups of cancer types. To address this issue, we exploited a cancer-specific aberrant transcription-induced chimeric RNA, designated A-Pas chiRNA, as a possible source of clinically relevant and targetable neoantigens. A-Pas chiRNA encodes a recently discovered cancer-specific chimeric protein that comprises full-length astrotactin-2 (ASTN2) C-terminally fused in-frame to the antisense sequence of the 18th intron of pregnancy-associated plasma protein-A (PAPPA). We used extracellular vesicles (EVs) from A-Pas chiRNA-transfected dendritic cells (DCs) to produce the cell-free anticancer vaccine DEXA-P . Treatment of immunocompetent cancer-bearing mice with DEXA-P inhibited tumour growth and prolonged animal survival. In summary, we demonstrate for the first time that cancer-specific transcription-induced chimeric RNAs can be exploited to produce a cell-free cancer vaccine that induces potent CD8+ T cell-mediated anticancer immunity. Our novel approach may be particularly useful for developing cancer vaccines to treat malignancies with low mutational burden or without mutation-based antigens. Moreover, this cell-free anticancer vaccine approach may offer several practical advantages over cell-based vaccines, such as ease of scalability and genetic modifiability as well as enhanced shelf life.

A prognostic nomogram for women with primary ovarian signet-ring cell carcinoma.

BACKGROUND: Primary ovarian signet-ring cell carcinoma (POSRCC) is a rare subtype of ovarian carcinoma that is characterized by abundant mucin accumulation. POSRCC is aggressive, and the prognostic factors associated with its clinical outcome remain poorly defined. This study aimed to elucidate the clinical characteristics and survival of patients with POSRCC, and to establish an effective prognostic nomogram and risk stratification model to predict the risks associated with patient outcomes. METHODS: Data of patients with POSRCC from the period 1975 to 2016 were collected from the Surveillance, Epidemiology, and End Results (SEER) database. Univariable and multivariable analyses of demographic factors, clinicopathological characteristics, and treatments were conducted to identify significant prognostic parameters. The identified independent variables were integrated to develop a nomogram and risk stratification model. The discrimination and calibration of the nomogram were assessed with the concordance index (C-index), receiver operating characteristic (ROC) curves, and calibration curves. RESULTS: A total of 172 patients were identified as being eligible to participate in this study. The median overall survival (OS) time was 7 months [95% confidence interval (CI), 4.6-9.4 months]. The 1-, 3-, and 5-year OS rates were 35.5%, 15.3%, and 6%, respectively. A multivariable analysis of the primary patients identified the independent predictors for survival as age at diagnosis, race, marital status, T (primary tumor size) stage, and chemotherapy, which were all incorporated into the nomogram. The C-index was 0.70 (95% CI, 0.66-0.75), which was statistically higher than that of the International Federation of Gynecology and Obstetrics (FIGO) staging system (0.58; 95% CI, 0.53-0.63). ROC curve analysis also showed that the nomogram had good discrimination, with an area under the curve (AUC) of 0.74, 0.62, and 0.71 for 1-, 3-, and 5-year survival, respectively. The calibration curves showed good agreement between the prediction by the nomogram and actual observations. A risk stratification model was further used to classify patients into a low-risk or high-risk group. The median OS time for the low- and high-risk groups was 13.0 months (95% CI, 9.33-16.67) and 2.0 months (95% CI, 1.12-2.89), respectively. Surgery did not significantly prolong survival in either group [low-risk group: hazard ratio (HR), 0.69; 95% CI, 0.45-1.07; P=0.09; high-risk group: HR, 0.55; 95% CI, 0.46-0.67; P=0.18]. CONCLUSIONS: The proposed nomogram and risk stratification model showed accurate prognostic prediction for POSRCC. These methods could improve individualized evaluations of survival and therapeutic decisions for patients with POSRCC.