Therapeutic cancer vaccines require robust cellular immunity for the efficient killing of tumor cells, and recent advances in neoantigen discovery may provide safe and promising targets for cancer vaccines. However, elicitation of T cells with strong antitumor efficacy requires intricate multistep processes that have been difficult to attain with traditional vaccination approaches. Here, a multifunctional nanovaccine platform has been developed for direct delivery of neoantigens and adjuvants to lymph nodes (LNs) and highly efficient induction of neoantigen-specific T cell responses. A PEGylated reduced graphene oxide nanosheet (RGO-PEG, 20−30 nm in diameter) is a highly modular and biodegradable platform for facile preparation of neoantigen vaccines within 2 h. RGO-PEG exhibits rapid, efficient (15−20% ID/g), and sustained (up to 72 h) accumulation in LNs, achieving >100-fold improvement in LN-targeted delivery, compared with soluble vaccines. Moreover, RGO-PEG induces intracellular reactive oxygen species in dendritic cells, guiding antigen processing and presentation to T cells. Importantly, a single injection of RGO-PEG vaccine elicits potent neoantigen-specific T cell responses lasting up to 30 days and eradicates established MC-38 colon carcinoma. Further combination with anti-PD-1 therapy achieved great therapeutic improvements against B16F10 melanoma. RGO-PEG may serve a powerful delivery platform for personalized cancer vaccination.

Osteosarcoma is the most common primary bone malignancy in children and young adults, but the role of adipose-derived mesenchymal stem cells (ADSCs) in the rapid progression of osteosarcoma is still unclear. Here, we found that ADSCs promoted tumour growth and invasion by increasing matrix metalloproteinase 2/9 (MMP2/9) expression in tumour cells. The persistent activation of signal transducer and activator of transcription 3 (STAT3) has been shown to directly promote tumour growth by mediating a wide spectrum of cellular responses, and STAT3 activation was detected in osteosarcoma cells co-cultured with ADSCs or treated with ADSC-conditioned medium. Furthermore, siRNA-mediated STAT3 inhibition in osteosarcoma cells decreased cell proliferation and invasion and down-regulated MMP2/9 expression. In addition, a nude mouse model of osteosarcoma was established by injecting luciferase-labelled MG63 cells into the tibia. As shown in in vivo bioluminescence images, ADSCs promoted tumour cell proliferation, invasion progression and metastasis. STAT3 inhibition attenuated tumour growth and metastasis and prolonged the survival of these mice. After the siRNA treatment, the MMP2, MMP9 and Ki67 levels decreased. Based on these data, stromal ADSCs promote osteosarcoma progression by increasing STAT3 signalling-mediated MMP2/9 expression.

Purpose. Adipose-derived mesenchymal stem cells (ADSCs) are increasingly applied in tendon repair. However, the underlying mechanisms of ADSC-derived extracellular vesicles (EVs) in tendon healing are largely unknown. In this study, we investigated the effect of the EVs secreted by ADSCs on the recovery of tendon injuries and its potential mechanism. Materials and Methods. We injected ADSCs into the injured tendon, followed by the evaluation of the tissue morphology, tenocyte proliferation, and oxidative stress. Then, the injured tenocytes were treated with EVs secreted by ADSCs, and oxidative stress and proliferation of tenocytes in vitro were detected. After the overexpression and knockdown of miR-19a and its target protein IGFBP3, the oxidative stress and proliferation of tenocytes in vitro were assessed. Finally, the injured tendon was treated with EVs, and the tissue morphology and proliferation of the injured tendon in vivo were examined. Results. ADSC-derived EVs were found to inhibit oxidative stress and promote proliferation of tenocytes isolated from an injury model of rats. EVs were shown to carry miR-19a which regulated the expression of IGFBP3 through binding to 3 ′ UTR of IGFBP3 mRNA. In addition, IGFBP3 promotes oxidative stress and inhibits proliferation of tenocytes. Finally, we found that ADSC-derived EVs promoted tendon wound healing in vivo. Conclusions. Our data suggest that treatment with ADSC-derived EVs ameliorates tendon injury by inhibiting oxidative stress and promoting proliferation in tenocytes. miR-19a carried by ADSC-derived EVs regulates IGFBP3 expression through binding to its 3 ′ UTR.

Quercus litseoides, an endangered montane cloud forest species, is endemic to southern China. To understand the genomic features, phylogenetic relationships, and molecular evolution of Q. litseoides, the complete chloroplast (cp) genome was analyzed and compared in Quercus section Cyclobalanopsis. The cp genome of Q. litseoides was 160,782 bp in length, with an overall guanine and cytosine (GC) content of 36.9%. It contained 131 genes, including 86 protein-coding genes, eight ribosomal RNA genes, and 37 transfer RNA genes. A total of 165 simple sequence repeats (SSRs) and 48 long sequence repeats with A/T bias were identified in the Q. litseoides cp genome, which were mainly distributed in the large single copy region (LSC) and intergenic spacer regions. The Q. litseoides cp genome was similar in size, gene composition, and linearity of the structural region to those of Quercus species. The non-coding regions were more divergent than the coding regions, and the LSC region and small single copy region (SSC) were more divergent than the inverted repeat regions (IRs). Among the 13 divergent regions, 11 were in the LSC region, and only two were in the SSC region. Moreover, the coding sequence (CDS) of the six protein-coding genes (rps12, matK, atpF, rpoC2, rpoC1, and ndhK) were subjected to positive selection pressure when pairwise comparison of 16 species of Quercus section Cyclobalanopsis. A close relationship between Q. litseoides and Quercus edithiae was found in the phylogenetic analysis of cp genomes. Our study provided highly effective molecular markers for subsequent phylogenetic analysis, species identification, and biogeographic analysis of Quercus.

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Soft-tissue allografts are inferior to hamstring tendon autografts with respect to subjective patient evaluation and knee stability but superior in the complication of hypoesthesia for patients undergoing primary ACL reconstruction.

Background: The biomechanical and tendon-bone incorporation properties of allograft-augmented hybrid grafts for anterior cruciate ligament (ACL) reconstruction compared with traditional autografts are unknown. Hypothesis: Using an autograft for ACL reconstruction yields better results on biomechanical testing, radiographic analysis, and histological evaluation versus using a hybrid graft. Study Design: Controlled laboratory study. Methods: A total of 66 adult male Sprague Dawley rats underwent unilateral ACL reconstruction with an autograft (AT group; n = 33) or a hybrid graft (HB group; n = 33). The grafts used in both groups were harvested from the peroneus longus tendon and were fixed by suturing to the surrounding periosteum. Samples were harvested for biomechanical testing, micro–computed tomography (CT), and histological evaluation at 4, 8, and 12 weeks postoperatively. Bone tunnels on the femoral and tibial sides were divided into 3 subregions: intra-articular (IA), midtunnel (MT), and extra-articular (EA). A cylinder-like volume of interest in the bone tunnel and a tubular-like volume of interest around the bone tunnel were used to evaluate new bone formation and bone remodeling, respectively, via micro-CT. Results: In the AT group, there were significantly higher failure loads and stiffness at 8 weeks (failure load: 3.04 ± 0.40 vs 2.09 ± 0.54 N, respectively; P = .006) (stiffness: 3.43 ± 0.56 vs 1.75 ± 0.52 N/mm, respectively; P < .001) and 12 weeks (failure load: 9.10 ± 1.13 vs 7.14 ± 0.94 N, respectively; P = .008) (stiffness: 4.45 ± 0.75 vs 3.36 ± 0.29 N/mm, respectively; P = .008) than in the HB group. With regard to new bone formation in the bone tunnel, in the AT group, the bone volume/total volume (BV/TV) was significantly higher than in the HB group on the tibial side at 8 weeks (IA: 22.21 ± 4.98 vs 5.16 ± 3.98, respectively; P < .001) (EA: 19.66 ± 7.19 vs 10.85 ± 2.16, respectively; P = .030) and 12 weeks (IA: 30.50 ± 5.04 vs 17.11 ± 7.31, respectively; P = .010) (MT: 21.15 ± 2.58 vs 15.55 ± 4.48, respectively; P = .041) (EA: 20.75 ± 3.87 vs 10.64 ± 3.94, respectively; P = .003). With regard to bone remodeling around the tunnel, the BV/TV was also significantly higher on the tibial side at 8 weeks (MT: 33.17 ± 8.05 vs 15.21 ± 7.60, respectively; P = .007) (EA: 25.19 ± 6.38 vs 13.94 ± 7.10, respectively; P = .030) and 12 weeks (IA: 69.46 ± 4.45 vs 47.80 ± 6.16, respectively; P < .001) (MT: 33.15 ± 3.88 vs 13.76 ± 4.07, respectively; P < .001) in the AT group than in the HB group. Sharpey-like fibers had formed at 8 weeks in the AT group. A large number of fibroblasts withdrew at 12 weeks. In the AT group, the width of the interface was significantly narrower at 4 weeks (85.86 ± 17.49 vs 182.97 ± 14.35 μm, respectively; P < .001), 8 weeks (58.86 ± 10.99 vs 90.15 ± 11.53 μm, respectively; P = .002), and 12 weeks (42.70 ± 7.96 vs 67.29 ± 6.55 μm, respectively; P = .001) than in the HB group. Conclusion: Using an autograft for ACL reconstruction may result in improved biomechanical properties and tendon-bone incorporation compared with a hybrid graft. Clinical Relevance: Augmenting small autografts with allograft tissue may result in decreased biomechanical performance and worse tendon-bone incorporation, increasing the risk of graft failure.

Key message Leaf symmetry and leaf size are explained by genetic variation between and within lineages and to a lesser extent by climatic factors, while leaf asymmetry can only be partly explained by geographic factors in Quercus aquifolioides Rehder & E.H. Wilson.Context Leaves are the primary photosynthetic organs of plants, and their morphology affects various crucial physiological processes potentially linked to fitness. Aims We explored the variation in leaf morphology of an alpine oak, Quercus aquifolioides, in order to examine its relationship to genetic, geographic, and climatic factors. Methods We conducted a genetic survey using 25 nuclear microsatellites. Based on Bayesian clustering analysis, 273 sampled trees from 29 populations of Q. aquifolioides were assigned to two lineages that correspond to the Western Sichuan Plateau-Hengduan Mountains (WSP-HDM) and Tibet geographic areas, with some individuals showing mixed ancestry. To undertake morphological analyses, we collected 1435 leaves from these trees and characterized them in terms of 13 landmarks. The metric dimensions of these leaves were digitally captured in the two-dimensional coordinates of these landmarks, then divided into leaf size and symmetric and asymmetric components of leaf shape. To analyze how different components of leaf morphology vary across lineages, we employed Procrustes Analysis of Variance (ANOVA), two-block partial least-square analysis (2B-PLS), and several other multivariate analysis approaches. We also applied distance-based redundancy analysis (dbRDAs) to explore relations between leaf morphology and genetic, geographic, and climatic factors. Results Multivariate analysis indicated significant differentiation in leaf symmetric shape components and leaf size between the WSP-HDM and Tibet lineages, while the mixed individuals were morphologically intermediate. The dbRDA analysis showed that most of the variation in symmetric components and leaf size was explained by genotypic effects, with the symmetric components of leaf shape being also significantly explained by geography and climate; however, variation in asymmetric components is only very weakly explained by geography. Conclusion Our results demonstrated that leaf morphological variation in shape and size across Q. aquifolioides geographic range is related to both its genetic differentiation and to a lesser extent to climatic factors. We discuss how these patterns could be interpreted in terms of both geographical isolations among and within lineages, and possible adaptive responses for particular traits, in contrast to asymmetric variation.