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Introduction TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) can induce apoptosis in a variety of cancer cells. However, drug resistance of tumor and short half-life seriously affects its clinical targeted therapy. Cabazitaxel (CTX) is a taxane drug, which can induce apoptosis or autophagy by inhibiting the phosphorylation of PI3K/Akt/mTOR and sensitive to some drug-resistant tumors. Therefore, we explored the possibility of developing a mesenchymal stem cell-derived exosomes (MSC-EXO) vector for oral squamous cell carcinoma (OSCC) to deliver CTX/TRAIL combinations. Methods After ultracentrifugation and dialysis, CTX/TRAIL loaded exosomes transfected MSC (MSCT)-derived exosome (EXO) (MSCT-EXO/CTX) were isolated and purified. The expression of CD63, CD9 and TRAIL was detected by BCA to confirm the origin of EXO. High-performance liquid chromatography (HPLC) was used to determine the drug loading of VPF and draw the in vitro release profile. MTT assay, flow cytometry and Western blot were used to detect the antitumor effect of MSCT-EXO/CTX in vitro. Subsequently, the antitumor effect of MSCT-EXO/CTX in vivo was verified by mouse model. Results The diameter of the membrane particles was about 60-150 nm. We have proved that the incorporation and release of CTX in MSCT-EXO can inhibit the activation of PI3K, Akt and mTOR, which is a possible synergistic mechanism of CTX. MSCT-EXO and CTX can induce the apoptosis of SCC25 tumor cells in a dose-dependent manner and exert a good synergistic effect in the proportion range of 10:1-5:1. The inherent activity of MSCT-EXO and the direct effect of MSCT-EXO/CTX on OSCC confirm that MSCT-EXO/CTX makes MSCT-EXO and CTX have an efficient synergistic effect and a highly effective pharmacological inhibition on cancer cells, as verified by the subsequent mouse model. MSCT-EXO/CTX showed the lowest relative tumor volume and the highest tumor inhibition rate (P Conclusion An MSCT-EXO-based CTX delivery system might be an effective anticancer method.

Purpose To evaluate the tear film stability after primary pterygium excision combined with Limbal stem cell transplantation (LSCT) or amniotic membrane transplantation (AMT). Methods We searched the PubMed, EMBASE, Web of Science, Cochrane Library, CNKI, Wan Fang and VIP databases for all studies on tear film stability after primary pterygium excision combined with LSCT or AMT. The mean difference (MD) and 95% confidence interval (CI) were calculated for outcomes using the fixed effect or random effect model.Results Seven studies with a total of 531 eyes were enrolled in our meta-analysis, which revealed that comparison between the LSCT group and the AMT group: Ocular Surface Disease Index (OSDI) 3 months postoperatively (MD=-5.16, 95%CI:-6.48,-3.85, P＜0.05), Tear break-up time (BUT) 1 or 3 months postoperatively (1 month: MD=0.30, 95%CI:-0.66,1.26, P=0.54; 3 months: MD=1.30, 95%CI:-0.13,2.72, P=0.07), Schirmer I test 1 or 3 months postoperatively (1 month: MD=0.05, 95%CI:-0.41,0.51, P=0.82; 3 months: MD=1.41, 95%CI:0.81,2.02, P＜0.05), Corneal fluorescein staining (CFS) score 1 month postoperatively (MD=-0.49, 95%CI:-1.29,0.31, P=0.23).Conclusion Primary pterygium excision combined with LSCT is associated with better tear film stability changes than AMT.

Several cryptic avian species have been validated by recent integrative taxonomic efforts in the Sino-Himalayan mountains, indicating that avian diversity in this global biodiversity hotspot may be underestimated. In the present study, we investigated species limits in the genus Tarsiger, the bush robins, a group of montane forest specialists with high species richness in the Sino-Himalayan region. Based on comprehensive sampling of all 11 subspecies of the six currently recognized species, we applied an integrative taxonomic approach by combining multilocus, acoustic, plumage and morphometric analyses. Our results reveal that the isolated north-central Chinese populations of Tarsiger cyanurus, described as the subspecies albocoeruleus but usually considered invalid, is distinctive in genetics and vocalisation, but only marginally differentiated in morphology. We also found the Taiwan endemic T. indicus formosanus to be distinctive in genetics, song and morphology from T. i. indicus and T. i. yunnanensis of the Sino-Himalayan mountains. Moreover, Bayesian species delimitation using BPP suggests that both albocoeruleus and formosanus merit full species status. We propose their treatment as 'Qilian Bluetail' T. albocoeruleus and 'Taiwan Bush Robin' T. formosanus, respectively.

Introduction: Lymphodepletion is a key factor for responses in both autologous and allogeneic cell therapies. We are currently evaluating in a Phase I trial, the combination of Temozolomide (TMZ) chemotherapy with methylguanine-DNA methyltransferase (MGMT) modified γδ T cells (Drug Resistant Immunotherapy, or DRI) during maintenance TMZ in patients receiving the Stupp protocol. In this report, we examine both DRI graft and circulating lymphocyte phenotype and function during the treatment phase. Methods: Patients meeting enrollment criteria undergo tumor resection, placement of a Rickham catheter in the tumor cavity, and an apheresis collection immediately prior to standard chemoradiotherapy (CRT). DRI follows TMZ on Day 1 of maintenance and consists of 150mg/m2 IV TMZ and concurrent intracranial injection of 1.0 x 107 DRI γδT cells. Peripheral blood (PB) was collected at apheresis and immediately prior to the start of each 28-day maintenance cycle. Flow cytometric (FACS) analysis of PB and DRI grafts using antibodies to CD3, CD4, CD8, CD16/56, CD19, TCR-γδ, Vδ1, Vδ2, CD197, CD27, CD28, CD57, CD45RA, Th1/Th2/Th17 PB serum cytokines, and DRI cytotoxic graft function were obtained. When applicable, FFPE sections of recurrent resected GBM tissues were examined for immune infiltration. Results: DRI grafts contained 74-93% activated γδ T cells with 0.3-0.5 MGMT copies/cell. Cytotoxicity against K562 ranged from 30% to 75% at 40:1 (E:T). Peripheral lymphodepletion was evident throughout TMZ treatment in the Stupp protocol. T cell counts fell precipitously after CRT and cycle 1 (1647+774 vs 609+214 cells/mL) and remained low throughout maintenance phase (range 214-1450 T cells/mL). NK counts remained normal and showed uneven recovery through the first three cycles but failed to recover for the remainder of maintenance phase. γδ T cells also recovered modestly through the first two cycles but failed to recover after additional cycles. Interestingly, CD45RA+CD27- effector T cells showed only slight increases for each patient throughout TMZ maintenance. Cytokine analysis did not show a clear trend except for consistent T cell expression of perforin and IP-10. One patient with recurrent GBM was successfully resected 110d following DRI. Histopathology revealed widespread necrosis with significant infiltration of CD4+ and CD8+ T cells and γδ T cells in the tumor parenchyma. Conclusions: Standard of care TMZ is lymphodepleting and DRI therapy can be conducted in an environment favorable to T cell persistence and sustained immune response. One treated patient demonstrated infiltration of both αβ and γδ T cells 110 days following a single dose of DRI γδ T cells. TMZ maintenance during the Stupp regimen prolongs lymphodepletion, thereby presenting a favorable setting for adoptive cell therapy. Citation Format: Lawrence S. Lamb, Lei Ding, Ryan C. Miller, Mariska ter Haak, Caitlyn Lucas, Becca Weekley, Samantha Youngblood, Cathy Langford, Guoling Chen, Louis B. Nabors. Maintenance-phase Temozolomide as a lymphodepletion platform for intracranialadoptiveγδ T cell-basedtherapy in primary high-grade gliomas [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 1173.

Both anthropogenic impacts and historical climate change could contribute to population decline and species extinction, but their relative importance is still unclear. Emerging approaches based on genomic, climatic and anthropogenic data provide a promising analytical framework to address this question. This study applied such an integrative approach to examine potential drivers for the endangerment of the green peafowl ( Pavo muticus ). Several demographic reconstructions based on population genomes congruently retrieved a drastic population declination since the mid-Holocene. Furthermore, a comparison between historical and modern genomes suggested genetic diversity decrease during the last 50 years. However, climate-based ecological niche models predicted stationary general range during these periods and imply the little impact of climate change. Further analyses suggested that human disturbance intensities were negatively correlated with the green peafowl's effective population sizes and significantly associated with its survival status (extirpation or persistence). Archaeological and historical records corroborate the critical role of humans, leaving the footprint of low genomic diversity and high inbreeding in the survival populations. This study sheds light on the potential deep-time effects of human disturbance on species endangerment and offers a multi-evidential approach in examining underlying forces for population declines.

The ongoing coronavirus disease 2019 (COVID-19) epidemic has made a huge impact on health, economies, and societies all over the world. Although reverse transcription-polymerase chain reaction (RT-PCR)-based nucleic acid detection has been primarily used in the diagnosis of COVID-19, it is time-consuming with limited application scenarios and must be operated by qualified personnel. Antibody test, particularly point-of-care antibody testing, is a suitable complement to nucleic acid test as it provides rapid, portable, and cost-effective detection of infections. In this study, a Rapid Antibody Test Kit was developed based on fluorescence immunochromatography for the sensitive, accurate, and automated detection of immunoglobulin M (IgM) and IgG antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in human serum, plasma, and whole blood samples within 10 min. The sensitivity, specificity, precision, and stability of the test kit were of good performance. No cross-activity and no interference was observed. In the multiple-center parallel study, 223 samples from hospitalized patients were used to evaluate the clinical specificity of the test. Both SARS-CoV-2 IgM and IgG achieved a clinical specificity of 98.21%. The clinical sensitivities of SARS-CoV-2 IgM and IgG were 79.54% and 87.45%, respectively, among 733 reverse transcription-polymerase chain reaction (RT-PCR) confirmed SARS-CoV-2 samples. For the combined IgM and IgG assays, the sensitivity and specificity were 89.22% and 96.86%, respectively. Our results demonstrate that the combined use of IgM and IgG could serve as a more suitable alternative detection method for patients with COVID-19, and the developed kit is of great public health significance for the prevention and control of the COVID-19 pandemic.

Endometrial carcinoma is one of the most common malignancies in the female reproductive system. Interleukin-37 (IL-37) is a newly discovered anti-inflammatory factor belonging to the IL-1 family. IL-37 has five different isoforms, and IL-37b is the most biologically functional subtype. In recent years, the protective roles of IL-37 in different cancers, including lung and liver cancers, have been successively reported. IL-37 also plays an important role in some gynecological diseases such as endometriosis, adenomyosis, and cervical cancer. However, the role and mechanism of IL-37b, especially the mature form of IL-37b, in endometrial carcinoma have not been elucidated. The present study demonstrated that IL-37 protein was downregulated in endometrial carcinoma cells compared with the control endometrium. IL-37b did not affect the proliferation and colony-forming ability of endometrial cancer cells. A mature form of IL-37b (IL-37bΔ1-45) effectively suppressed the migration and invasion of endometrial cancer cells by decreasing the expression of matrix metalloproteinase 2 (MMP2) via Rac1/NF-κB signal pathway. However, it did not affect epithelial-mesenchymal transition (EMT) or filamentous actin (F-actin) depolymerization of endometrial cancer cells. IL-37bΔ1-45 attenuated tumor metastasis in a peritoneal metastatic xenograft model of endometrial cancer. To sum up, these results suggested IL-37b could be involved in the pathogenesis of endometrial carcinoma and provide a novel target for the diagnosis and treatment of endometrial carcinoma.

Background The White-browed Shortwing (Brachypteryx montana) is widespread from the central Himalayas to the southeast Chinese mainland and the island of Taiwan, the Philippines and Indonesia. Multiple subspecies are recognised, and several of these have recently been suggested to be treated as separate species based on differences in morphology and songs. Methods We here analyse plumage, morphometrics, songs, two mitochondrial and two nuclear markers, and geographical distributions of the two mainland Asian taxa B. m. cruralis and B. m. sinensis and the Taiwanese B. m. goodfellowi. Results We conclude that these differ congruently in morphology, songs and DNA. Male B. m. goodfellowi is the most divergent in plumage (sexually monomorphic, unlike the two others; male similar to female), and B. m. cruralis and B. m. sinensis differ in male plumage maturation. The song of B. m. cruralis is strongly divergent from the others, whereas the songs of B. m. sinensis and B. m. goodfellowi are more similar to each other. Brachypteryx m. sinensis and B. m. goodfellowi are sisters, with an estimated divergence time 4.1 million years ago (mya; 95% highest posterior distribution [HPD] 2.8–5.5 mya), and B. m. cruralis separated from these two 5.8 mya (95% HPD 4.1–7.5 mya). We also report notable range extensions of B. m. sinensis as well as sympatry between this taxon and B. m. cruralis in Sichuan Province, China. Brachypteryx m. montana from Java is found to be more closely related to Lesser Shortwing (B. leucophris) and Rusty-bellied Shortwing (B. hyperythra) than to the mainland Asian and Taiwanese taxa. Conclusion Our data support a recent proposal to treat the three mainland Asian and Taiwanese taxa as three species, separate from B. montana sensu stricto: B. cruralis (central Himalayas to south central China and south Vietnam), B. sinensis (north central to southeastern part of mainland China) and B. goodfellowi (Taiwan Island).

SummaryThe arms race between pathogens and the immune system of their hosts is a critical evolutionary force that affects the ecology and life history of organisms. An increased infection risk selects for adaptations in immune genes that encode receptors involved in pathogen recognition and the initiation of innate immune responses, including Toll-like receptor (TLR) genes. Although recent studies assessed the evolution of major histocompatibility complex (MHC) genes, the ecological and evolutionary processes that drive the evolution of immune genes across major phylogenetic lineages remain unstudied. Moreover, trade-offs between immune responses and other energy-demanding vital functions may limit the resource allocation into costly immune functions, and therefore affect the evolution of immune genes. Here we assess the evolutionary patterns of six TLR genes across 121 bird species, covering 95% of extant orders that represent diverse ecologies and life histories. Selection analyses revealed that all six TLR genes show strong signs of purifying selection, while few sites underwent episodic positive selection. Comparative phylogenetic analyses showed that the intensity of positive selection of TLR genes is associated with long-distance migration, extensive parental care (i.e., altricality and prolonged parent-offspring association), and a large body size (a proxy of increased longevity). Together, these results suggest that the evolution of immune genes is characterized by episodic positive selection, and is shaped by an increased inflection risk and extensive parental care that buffers the costs of immune functions.

We collected multi‐locus data to test whether hybridization has occurred between a critically endangered waterbird, the Chinese Crested Tern Thalasseus bernsteini, and the Greater Crested Tern Thalasseus bergii. Phylogenetic analysis shows these two tern species are sister species, having diverged

The Chinese Crested Tern, Thalasseus bernsteini, is one of the most endangered seabird species in the world, with only small breeding populations of around 100 individuals. To profile the genetic properties of this population, we screened 200 short tandem repeat sequences from a de novo assembled genome of the Great Crested Tern, T. bergii, which is the sister species of T. bernsteini. We designed a panel of 12 novel microsatellites, and genotyped nine and 23 individuals of T. bernsteini and T. bergii, respectively. The results showed that T. bernsteini genetic diversity was lower than that of T. bergii, and no inbreeding was detected. There was no sign of recent bottleneck events in the two species, but T. bernsteini showed signals of a post-bottleneck population expansion. Gene flow analysis indicated introgression between the two species, but the magnitude from T. bernsteini to T. bergii was higher than the opposite. This could be explained by Hubb’s principle, which states that there is a propensity of rarer species to reproduce with abundant congeners. Overall, our efforts provide useful genetic tools for conservation genetic studies of T. bernsteini and potentially other crested terns which face several natural and anthropogenic threats. The findings of this study further highlight that introgression may be a neglected factor in species conservation that needs to be seriously considered for conservation management. Availability of data and materials The dataset of all microsatellite are available from the corresponding author upon request.

Our knowledge of the systematics and taxonomy of Asian birds has improved much in the last two decades, and the number of recognised species has increased significantly as a result of in-depth studies using an integrative taxonomic approach. The Sino-Himalayan mountains harbor a high level of passerine diversity. Several allopatric or parapatric taxa that are currently treated as subspecies of polytypic species within that region are likely to deserve full species status, and thus their taxonomic status needs to be revisited. Based on analyses of multilocus data, vocalizations and morphology, we propose that the white-tailed rubythroat Calliope pectoralis should be treated as two species, the Himalayan rubythroat C. pectoralis sensu stricto in the Tian Shan and Himalayan mountains, and the Chinese rubythroat C. tschebaiewi in the mountains of southwestern and north-central China. According to our dating analyses based on mitochondrial loci, these two species diverged approximately 2.2 million yr ago. We further found that C. tschebaiewi was paraphyletic to C. pectoralis sensu stricto in nuclear data, which demonstrates a state of mitonuclear discordance that warrants further work. Our results suggest that geographic changes and glacial cycles in the Pleistocene may have caused allopatric divergence in the C. pectoralis complex. Our study stresses the importance of applying an integrative taxonomy approach to fully unravel the true avian diversity in the Sino-Himalayan Mountains.

This study investigates the functioning mechanisms of how high performance work systems (HPWS) affect organizational performance. We propose that (HPWS) can positively affect organizational performance through the mediating role of entrepreneurial orientation. An organization with high performance work systems can perform better if it enjoys high level of organizational learning. We design and administer a survey questionnaire to high-level executives or founders of companies from manufacturing and service industries and receive 176 valid responses. The results of the empirical data indicate that the relationship between high performance work systems and corporate performance is more positive when organizational learning is stronger. Entrepreneurial orientation partially mediates the relationship between high performance work systems and organizational performance. This study opens new research avenues by extending and incorporating explanations and predictions of HPWS and entrepreneurial orientation, two areas that largely have been considered independently of each other. Implications for practice and directions for future research are provided.

Diversity and distributions of cryptic species have long been a vexing issue. Identification of species boundaries is made difficult by the lack of obvious morphological differences. Here, we investigate the cryptic diversity and evolutionary history of an underappreciated group of Asian frog species (Megophrys) to explore the pattern and dynamic of amphibian cryptic species. We sequenced four mitochondrial genes and five nuclear genes and delineated species using multiple approaches, combining DNA and mating-call data. A Bayesian species tree was generated to estimate divergence times and to reconstruct ancestral ranges. Macroevolutionary analyses and hybridization tests were conducted to explore the evolutionary dynamics of this cryptic group. Our phylogenies support the current subgenera. We revealed 43 cryptic species, 158% higher than previously thought. The species-delimitation results were further confirmed by mating-call data and morphological divergence. We found that these Asian frogs entered China from the Sunda Shelf 48 Mya, followed by an ancient radiation event during middle Miocene. We confirmed the efficiency of the multispecies coalescent model for delimitation of species with low morphological diversity. Species diversity of Megophrys is severely underappreciated, and species distributions have been misestimated as a result.

Additional file 6: Fig. S1. Phylogeny of the continental and Taiwanese Brachypteryx montana taxa, with B. m. montana, B. leucophris carolinae and B. hyperythra as outgroups, based on BEAST analysis of the mitochondrial cytochrome b and ND2 and nuclear myoglobin and ODC introns analysed in four partitions. Includes three cruralis samples not included in Fig. 1 due to missing sequences. Fig. S2. Trees inferred by BEAST based on cytb and ND2 (a) and myo and ODC (b), each dataset analysed in two partitions; and single-locus analyses of myo (c) and ODC (d).

A new species, Gracixalus jinggangensis sp. nov., is described based on a series of specimens collected from Mount Jinggang, Jiangxi Province, southeastern China. The new species is distinguished from all other known congeners by the following combination of morphological characters: relatively small body size, SVL 27.9–33.8 mm in nine males and 31.6 mm in a single female; upper eyelid and dorsum lacking spines; skin of dorsal and lateral surface of head, body and limbs rough with sparsely scattered with tubercles; ventral skin granular; tibiotarsal projection absent; finger webbing rudimentary; toes with moderately developed webbing; brown to beige above in life, with an inverse Y-shaped dark brown marking extending from the interorbital region to the middle of dorsum; males with a single, subgular vocal sac, barely visible nuptial pads with minute granules on the dorsal surface of the bases of first and second fingers. The new species is also genetically divergent from all other Gracixalus for which comparable 16S rRNA gene sequences are available. The discovery of Gracixalus jinggangensis sp. nov. represents the twelfth known species in the genus.

Objective The study aimed to investigate whether Foxp3-expressing sensitized Teff cells could inhibit allograft rejection in corneal allograft transplantation mouse model. Methods Foxp3-expressing sensitized Teff cells were constructed by transfection of retroviral expression plasmid expressing Foxp3 into the sensi-Teff cells from a Balb/c mouse immunized by C57BL/6(H2b) mouse splenocytes. Balb/c mice were randomly divided into 5 groups: Four groups received tail vein injection of Foxp3-expressing sensitized Teff cells, or Foxp3-expressing Teff cells, or Treg cells or no intervention 1 day prior to corneal allograft transplantation. C57BL/6(H2b) was the donor mouse. The last group received corneal autograft transplantation. Corneal allograft survival time and percentage of CD4 + T cells were detected. ELISPOT and Footpad swelling test were used to measure IL-2 and IFN-γ, and delayed-type hypersensitivity (DTH) response, respectively. Results Mice that had received an injection of Foxp3-expressing sensitized T cells prior to an allograft corneal transplantation, showed significantly longer survival time of corneal allograft, decreased percentage of CD4 + T cells, IL-2 and IFN-γ, and alleviated footpad swelling than the mice that had received either Foxp3-Teff or Treg cells. Conclusion Foxp3-sensi-Teff cell treatment that prolongs corneal allograft survival in the mouse model, might partly through suppressing CD4 + T cells, IL-2 and IFN-γ.

Species not only responded idiosyncratically to past climate changes, there were also regionally contrasting effects on spatio-temporal diversification patterns. Studies of closely related species appear to be a particularly promising comparative approach to disentangle such regionally differential impacts. In this study, we undertook a comprehensive geographic sampling to investigate the evolutionary history of the Holarctic Sand Martin Riparia riparia and the chiefly Central and East Asian Pale Martin R. diluta. Previous phylogenetic studies using only a limited geographic sampling, particularly for the latter, revealed the two to be genetically distinct, with the former showing only a shallow genetic structure in mitochondrial DNA. Based on one mitochondrial, one autosomal and one Z-linked nuclear marker, we confirmed the shallow genetic structure in R. riparia even when including the morphologically relatively distinct subspecies R. r. shelleyi from the Nile Valley in Egypt and probably the Middle East. On the other hand the different subspecies of R. diluta, i.e. R. d. diluta from Central Asia, R. d. indica from the northwestern Indian Subcontinent, R. d. tibetana from the Tibetan Plateau and R. d. fohkienensis from southeastern China, were found to be genetically distinct. Their diversification started before the Early to Middle Pleistocene Transition, which was followed by a pronounced succession of glacial and interglacial periods. These rather old divergence events contrast with the lack of any strong phylogeographic structure in R. riparia. Strongly structured populations and regional diversification have been reported in different forest passerine families of South-East Asia. Here we demonstrate, however, that species characteristic of open-country habitats such as R. diluta might display a similar pattern. Morphometric analyses of 120 individuals revealed no clear differences between the different subspecies of R. diluta. Given their similarity also in plumage features, we refrain from proposing any splits despite their marked genetic differentiation, pending further studies and particularly the discovery of potential secondary contact zones.

The big-headed turtle (Platysternon megacephalum) is critically endangered because of overharvesting, illegal trade, and habitat destruction. Assessment of genetic variability in existing populations becomes very important to the taxonomy and conservation of this species. Here we describe 14 microsatellite loci isolated from an enriched genomic library of the big-headed turtle, and the polymorphisms of these loci were assessed in 28 individuals from Huizhou, Heyuan, Zhaoqing, and Shaoguan of Guangdong, China. The range of polymorphism information content is 0.305-0.738, and no evidence of significant linkage disequilibrium was found among any pairs of loci. These 14 new polymorphic microsatellite loci can be used in population genetics, taxonomy, phylogeography, behavior ecology, and conservation efforts of Platysternon megacephalum.

Objective To investigate the antitumor effect of endostatin combined with tumor antigen-pulsed dendritic cell (DC)-T cell therapy on lung cancer. Methods Transplanted Lewis lung cancer (LLC) models of C57BL/6 mice were established by subcutaneous injection of LLC cells in left extremity axillary. Tumor antigen-pulsed DC-T cells from spleen cells and bone of mice were cultured in vitro. Tumor-bearing mice were randomly divided into three groups, including DC-T+endostatin group, DC-T group, and phosphate-buffered saline (PBS) control group. Microvessel density (MVD) of tumor tissue in tumor-bearing mice was determined by immunohistochemistry (IHC). The expressions of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α (HIF-1α) were determined by Western blotting and IHC staining. The proportions of CD8+ T cells, mature dendritic cells (mDC), tumor-associated macrophages [TAM (M1/M2)], and myeloid-derived suppressor cells (MDSC) in suspended cells of tumor tissue were determined by flow cytometry. The expressions of interleukin (IL)-6, IL-10, IL-17, transforming growth factor-β (TGF-β) and interferon-γ (IFN-γ) in suspended cells of tumor tissue were detected by enzyme-linked immune sorbent assay (ELISA). Results DC-T cells combined with endostatin remarkably suppressed tumor growth. MVD of mice in DC-T+endostatin group was significantly lower than that of the control group and DC-T monotherapy group. The expressions of VEGF, IL-6 and IL-17 in tumors were markedly decreased, but IFN-γ and HIF-1α increased after treating with DC-T cells combined with endostatin, compared to control group and DC-T group. In the DC-T+endostatin group, the proportions of MDSC and TAM (M2 type) were significantly decreased, mDC and TAM (M1 type) were up-regulated, and CD8+ T cells were recruited to infiltrate tumors, in contrast to PBS control and DC-T monotherapy. DC-T cells combined with endostatin potently reduced the expressions of IL-6, IL-10, TGF-β and IL-17 in tumor tissue, and enhanced the expression of IFN-γ. Conclusions The study indicated the synergic antitumor effects between endostatin and tumor antigen-pulsed DC-T cells, which may be a prospective therapy strategy to achieve potent antitumor effects on lung cancer.

Endostatin has previously been demonstrated to efficiently inhibit the angiogenesis and growth of endothelial cells. However, the role of endostatin in the tumor microenvironment remains to be elucidated. To investigate the antitumor effect of endostatin in lung cancer, the present study was designed to explore the alterations of microvessel density in Lewis lung cancer models and the expression of vascular endothelial growth factor (VEGF), interleukin (IL)-6, IL-17, interferon (IFN)-γ and hypoxia inducible factor (HIF)-1α, following endostatin therapy. It was demonstrated that the growth and angiogenesis of tumors were markedly suppressed by treatment with endostatin, compared with control group. The microvessel density in mice treated with endostatin was significantly inhibited in a dose-dependent manner. The expression levels of VEGF, IL-6 and IL-17 in tumors were decreased, however IFN-γ and HIF-1α expression levels were increased, following treatment with endostatin. In addition, the proportion of myeloid derived suppressor cells and tumor associated macrophages (TAMs; M2 type) were significantly decreased, whereas those of mature dendritic cells and TAMs (M1 type) were increased, and cluster of differentiation (CD)8+ T cells were recruited to infiltrate the tumors following treatment with endostatin. In addition, the expression levels of IL-6, IL-10, tumor growth factor-β and IL-17 in tumor tissue were potently decreased with endostatin therapy. These results indicated that endostatin efficiently inhibited tumor angiogenesis and reversed the immunosuppressive microenvironment associated with the presence of tumors.

Bag-of-features based approaches have become prominent for image retrieval and image classification tasks in the past decade. Such methods represent an image as a collection of local features, such as image patches and key points with scale invariant feature transform (SIFT) descriptors. To improve the bag-of-features methods, we first model the assignments of local descriptors as contribution functions, and then propose a novel multiple assignment strategy. Assuming the local features can be reconstructed by their neighboring visual words in a vocabulary, reconstruction weights can be solved by quadratic programming. The weights are then used to build contribution functions, resulting in a novel assignment method, called quadratic programming (QP) assignment. We further propose a novel visual word weighting method. The discriminative power of each visual word is analyzed by the sub-similarity function in the bin that corresponds to the visual word. Each sub-similarity function is then treated as a weak classifier. A strong classifier is learned by boosting methods that combine those weak classifiers. The weighting factors of the visual words are learned accordingly. We evaluate the proposed methods on medical image retrieval tasks. The methods are tested on three well-known data sets, i.e., the ImageCLEFmed data set, the 304 CT Set, and the basal-cell carcinoma image set. Experimental results demonstrate that the proposed QP assignment outperforms the traditional nearest neighbor assignment, the multiple assignment, and the soft assignment, whereas the proposed boosting based weighting strategy outperforms the state-of-the-art weighting methods, such as the term frequency weights and the term frequency-inverse document frequency weights.

The authors report here the clinical, genetic, molecular and biochemical characterisation of a large five-generation Han Chinese pedigree with maternally transmitted non-syndromic hearing loss. 17 of 35 matrilineal relatives exhibited variable severity and age at onset of sensorineural hearing loss. The average age at onset of hearing loss in matrilineal relatives of this family is 29 years, while matrilineal relatives among families carrying other mitochondrial DNA mutations developed hearing loss with congenital conditions or early age at onset. Molecular analysis of their mitochondrial genome identified the novel heteroplasmic T12201C mutation in the transfer RNA (tRNA)(His) gene. The levels of T12201C mutation in matrilineal relatives of this family correlated with the severity and age at onset of non-syndromic hearing loss. By contrast, other heteroplasmic mitochondrial DNA mutations often cause syndromic hearing loss. The T12201C mutation destabilises a highly conservative base-pairing (5A-68U) on the acceptor stem of tRNA(His). tRNA northern analysis revealed that the T12201C mutation caused an ∼75% reduction in the steady-state level of tRNA(His). An in vivo protein labeling analysis showed an ∼47% reduction in the rate of mitochondrial translation in cells carrying the T12201C mutation. Impaired mitochondrial translation is apparently a primary contributor to the marked reduction in the rate of overall respiratory capacity, malate/glutamate-promoted respiration, succinate/glycerol-3-phosphate-promoted respiration or N,N,Ń,Ń-tetramethyl-p-phenylenediamine/ascorbate-promoted respiration. These data provide the first direct evidence that mitochondrial dysfunctions caused by the heteroplasmic tRNA(His) mutation lead to late-onset non-syndromic deafness. Thus, the authors' findings provide new insights into the understanding of pathophysiology and valuable information on the management and treatment of maternally inherited hearing loss.

Objective To investigate the effect of emodin on lipopolysaccharides (LPS)-induced corneal injury in rats. Methods Three parallel incisions on the central surface of corneal epithelium were made and LPS was applied on them to induce corneal injury in Wistar rats. All rats were randomly divided into emodin group ( n =40) and keratitis group ( n =40). Rats in the emodin group received subconjunctival injection of emodin and rats in the keratitis group received its vehicle 30 minutes before LPS exposure. At different time points—1, 3, 6, 12, and 24 hours after LPS exposure, the symptoms of all rats were observed and the severity of their ocular inflammation was examined with a slit lamp microscope, then 8 rats in each group were killed through cervical dislocation and their eyes were enucleated and prepared to observe pathological changes of corneal tissue under a light microscope. The activation of nuclear factor-κB (NF-κB) under different conditions was determined by Western blot. Immunocytochemistry staining with an antibody against intercellular adhesion molecule-1 (ICAM-1) was performed to identify positive cells in corneal tissues. Results The model of acute keratitis was successfully established in Wistar rats. LPS could induce a typical corneal inflammatory response, such as hyperemia, corneal edema and opacity, which were observed in model rats. Compared with keratitis group, both ocular behaviors and damages of the corneal structure were improved in emodin group. Furthermore, the activation of NF-κB and the expression of ICAM-1 induced by LPS were markedly inhibited in emodin group. Conclusion Emodin can inhibit the activation of NF-κB and the expression of ICAM-1 induced by LPS in corneas, protect against acute corneal injury, and improve symptoms in rats.

Although there have been many experimental studies demonstrating that bone marrow-derived mesenchymal stem cells (MSCs) have the potential to differentiate into mesenchymal tissues such as osteocytes, chondrocytes, and adipocytes in vivo and in vitro, little information is available regarding their potential to differentiate into lymphatic endothelial cells. Therefore, we chose to investigate differentiation of MSCs into lymphatic endothelial cells using stimulation with members of the vascular endothelial growth factor (VEGFs) family. Rat MSCs were isolated from bone marrow aspirate of Sprague-Dawley rats as previously described and characterized with flow cytometry for surface markers CD14, CD34, CD29, and CD90. Purified MSCs were plated and cultured in the presence of VEGF-A, VEGF-C, or the combination of both for 10 days. We examined the cells for Prox-1 and LYVE-1 by immunocytochemistry, RT-PCR, and Western blot analysis. Results demonstrated that compared to controls, cell differentiated with VEGF-A, VEGF-C and VEGF-A+VEGF-C expressed Prox-1 and LYVE-1. Our results indicate that MSCs induced by VEGFs are capable of differentiating into lymphatic endothelial-like cells in vitro, and this response has the potential to make them attractive candidates for the development of autologous tissue grafts for future therapy.

Cell cycle re-entry by cochlear supporting cells and/or hair cells is considered one of the best approaches for restoring hearing loss as a result of hair cell damage. To identify mechanisms that can be modulated to initiate cell cycle re-entry and hair cell regeneration, we studied the effect of activating the sonic hedgehog (Shh) pathway. We show that Shh signaling in postnatal rat cochleae damaged by neomycin leads to renewed proliferation of supporting cells and hair cells. Further, proliferating supporting cells are likely to transdifferentiate into hair cells. Shh treatment leads to inhibition of retinoblastoma protein (pRb) by increasing phosphorylated pRb and reducing retinoblastoma gene transcription. This results in upregulation of cyclins B1, D2, and D3, and CDK1. These results suggest that Shh signaling induces cell cycle re-entry in cochlear sensory epithelium and the production of new hair cells, in part by attenuating pRb function. This study provides an additional route to modulate pRb function with important implications in mammalian hair cell regeneration.

Bag-of-features based approaches have become prominent for image retrieval and image classification tasks in the past decade. Such methods represent an image as a collection of local features, such as image patches and key points with scale invariant feature transform (SIFT) descriptors. To improve the bag-of-features methods, we first model the assignments of local descriptors as contribution functions, and then propose a novel multiple assignment strategy. Assuming the local features can be reconstructed by their neighboring visual words in a vocabulary, reconstruction weights can be solved by quadratic programming. The weights are then used to build contribution functions, resulting in a novel assignment method, called quadratic programming (QP) assignment. We further propose a novel visual word weighting method. The discriminative power of each visual word is analyzed by the sub-similarity function in the bin that corresponds to the visual word. Each sub-similarity function is then treated as a weak classifier. A strong classifier is learned by boosting methods that combine those weak classifiers. The weighting factors of the visual words are learned accordingly. We evaluate the proposed methods on medical image retrieval tasks. The methods are tested on three well-known data sets, i.e., the ImageCLEFmed data set, the 304 CT Set, and the basal-cell carcinoma image set. Experimental results demonstrate that the proposed QP assignment outperforms the traditional nearest neighbor assignment, the multiple assignment, and the soft assignment, whereas the proposed boosting based weighting strategy outperforms the state-of-the-art weighting methods, such as the term frequency weights and the term frequency-inverse document frequency weights.

Given their recent worldwide declines and extinctions, characterization of species-level diversity is of critical importance for large-scale biodiversity assessments and conservation of amphibians. This task is made difficult by the existence of cryptic species complexes, species groups comprising closely related and morphologically analogous species. The combination of morphology, genetic, and bioacoustic analyses permits robust and accurate species identification. Using these methods, we discovered two undescribed Xenophrys species, namely Xenophrys lini sp. nov. and Xenophrys cheni sp. nov. from the middle range of Luoxiao Mountains, southeast China. These two new species can be reliably distinguished from other known congeners by morphological and morphometric differences, distinctness in male advertisement calls, and substantial genetic distances (>3.6%) based on the mitochondrial 16s and 12s rRNA genes. The two new species, together with X. jinggangensis, are sympatric in the middle range of Luoxiao Mountains but may be isolated altitudinally and ecologically. Our study provides a first step to help resolve previously unrecognized cryptic biodiversity and provides insights into the understanding of Xenophrys diversification in the mountain complexes of southeast China.

Background The 1555A > G mutation is the most common cause of aminoglycoside-induced and non-syndromic deafness. However, the variable clinical phenotype and incomplete penetrance of A1555G-induced hearing loss complicate our understanding of this mutation. Environmental factors, nuclear genes, mitochondrial haplotypes/variants and a possible threshold effect have been reported to may be involved in its manifestation. Methods Here, we performed a clinical, molecular, genetic and phylogenic analysis in a six-generation Chinese family. Results A clinical evaluation revealed that affected individuals without aminoglycoside exposure developed hearing loss extending gradually from 12000 Hz to 8000 Hz and then to 4000 Hz. Using pyrosequencing, we detected an identical homoplasmic 1555A > G mutation in all individuals except one. We did not find any correlation between the mutation load and the severity of hearing loss. T123N coexisted with the 1555A > G mutation in six affected subjects in our pedigree. Analysis of the complete mtDNA genome of this family revealed that this family belonged to haplotype B4C1C and exhibited high penetrance. Upon the inclusion of subjects that had been exposed to aminoglycosides, the penetrance of the hearing loss was 63.6%.; without exposure to aminoglycosides, it was 51.5%. This pedigree and another reported Chinese pedigree share the same haplotype (B4C1C) and lack functionally significant mitochondrial tRNA variants, but nevertheless they exhibit a different penetrance of hearing loss. Conclusions Our results imply that the factors responsible for the higher penetrance and variable expression of the deafness associated with the 1555A > G mutation in this pedigree may not be mtDNA haplotype/variants, but rather nuclear genes and/or aminoglycosides.