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Metabolic programming is deeply intertwined with early embryonic development including zygotic genome activation (ZGA), the polarization of zygotic cells, and cell fate commitment. It is crucial to establish a noninvasive imaging technology that spatiotemporally illuminates the cellular metabolism pathways in embryos to track developmental metabolism in situ. In this study, we used two high‐quality genetically encoded fluorescent biosensors, SoNar for NADH/NAD+ and iNap1 for NADPH, to characterize the dynamic regulation of energy metabolism and redox homeostasis during early zygotic cleavage. Our imaging results showed that NADH/NAD+ levels decreased from the early to the late two‐cell stage, whereas the levels of the reducing equivalent NADPH increased. Mechanistically, transcriptome profiling suggested that during the two‐cell stage, zygotic cells downregulated the expression of genes involved in glucose uptake and glycolysis, and upregulated the expression of genes for pyruvate metabolism in mitochondria and oxidative phosphorylation, with a decline in the expression of two peroxiredoxin genes, Prdx1 and Prdx2. Collectively, with the establishment of in situ metabolic monitoring technology, our study revealed the programming of redox metabolism during ZGA. [ABSTRACT FROM AUTHOR]

Long non-coding RNAs (lncRNAs) regulate a series of physiological processes and play an important role in development, metabolism and disease. Our previous studies showed that lncRNAs involved in skeletal muscle differentiation. Here, we demonstrated that lncRNA Has2os is highly expressed in skeletal muscle and significantly elevated during skeletal cell differentiation. The knockdown of Has2os inhibited myocyte fusion and impeded the expression of the myogenic factors MyHC and Mef2C. Mechanically, Has2os regulates skeletal muscle differentiation by inhibiting the JNK/MAPK signaling pathway. Furthermore, we also revealed that Has2os is involved in the early stage of regeneration after muscle injury, and the JNK/MAPK signaling pathway is activated at both protein and mRNA levels during early repair. Our results demonstrate the new function of lncRNA Has2os, which plays crucial roles during skeletal muscle differentiation and muscle regeneration, providing a basis for the therapy of lncRNA-related muscle diseases. [ABSTRACT FROM AUTHOR]

Breast cancer is the most common cancer diagnosed in women. Breast cancer research is currently based mainly on animal models and traditional cell culture. However, the inherent species gap between humans and animals, as well as differences in organization between organs and cells, limits research advances. The breast cancer organoid can reproduce many of the key features of human breast cancer, thereby providing a new platform for investigating the mechanisms underlying the development, progression, metastasis and drug resistance of breast cancer. The application of organoid technology can also promote drug discovery and the design of individualized treatment strategies. Here, we discuss the latest advances in the use of organoid technology for breast cancer research. [ABSTRACT FROM AUTHOR]

TIGAR (TP53‐induced glycolysis and apoptosis regulator) is a p53‐inducible gene and its expression resulted in controlling metabolism and protection from apoptosis. Furthermore, TIGAR participated in promoting the pentose phosphate pathway and help in lowering intracellular reactive oxygen species. miR‐885‐5p has also been reported to be involved in liver tumorigenesis, but whether miR‐885‐5p has a regulatory effect on TIGAR expression is unknown. In this study, we found that their levels were correlated to each other and positively related to cell malignancy. Exogenous miR‐885‐5p induced TIGAR expression through a p53‐independent pathway. The promoter region of TIGAR harbors two tandem putative miR‐885‐5p target sites. Cotransfection of synthetic miR‐885 with TIGAR promoter reporter constructs significantly enhanced TIGAR promoter activity via binding with target sites. Furthermore, miR‐885‐5p and its precursor pre‐miR‐885 had the same stimulatory impact on TIGAR expression. Chromatin immunoprecipitation analysis further verified that increased miR‐885‐5p potentiated the accessibility of TIGAR promoter chromatin to transcriptional factors and facilitated TIGAR expression. miR‐885‐5p and its precursor both can interact mechanically with TIGAR promoter binding site and alter local chromatin structure, and subsequently upregulate TIGAR expression and participate in liver tumorigenesis. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs/miRs) show great promise as novel cancer biomarkers. Several studies have revealed an association between abnormal miRNA expression and the risk of various cancer types. However, the diagnostic accuracy and reliability of miRNAs remains unclear. The present meta-analysis was performed to summarize the overall diagnostic performance of miR-195 for cancer. The PubMed, Cochrane Library, Wanfang and China National Knowledge Infrastructure databases were searched for associated literature published until December 10, 2017. Eligible studies were selected using multiple search strategies based on study selection criteria. Measures, including sensitivity and specificity, of the performance of miR-195 as a cancer diagnostic tool were pooled using bivariate meta-analysis models. All analyses were performed using Stata 14.0. The pooled analysis included 8 studies comprising 735 cases and 547 controls. The pooled diagnostic results calculated from all studies were as follows: Sensitivity, 0.79 [95% confidence interval (CI), 0.69–0.87]; specificity, 0.84 (95% CI, 0.68–0.93); positive likelihood ratio, 4.9 (95% CI, 2.50–9.50); negative likelihood ratio, 0.25 (95% CI, 0.18–0.35); diagnostic odds ratio, 20 (95% CI, 10.00–38.00); and area under the curve, 0.87 (95% CI, 0.84–0.90). Deeks' funnel plot asymmetry test suggested no potential publication bias (P=0.53). The present meta-analysis indicated that miR-195 could be a reliable non-invasive biomarker for the diagnosis of cancer. Further large-scale prospective studies are necessary to confirm the present findings and the clinical value of miR-195 for future diagnostics. [ABSTRACT FROM AUTHOR]

Purpose: Breast cancer is a heterogeneous disease usually including four molecular subtypes such as luminal A, luminal B, HER2-enriched, and triple-negative breast cancer (TNBC). TNBC is more aggressive than other breast cancer subtypes. Despite major advances in ER-positive or HER2-amplified breast cancer, there is no targeted agent currently available for TNBC, so it is urgent to identify new potential therapeutic targets for TNBC. Methods: We first used microarray analysis to compare gene expression profiling between TNBC and non-TNBC. Furthermore an integrated analysis was conducted based on our own and published data, leading to more robust, reproducible and accurate predictions. Additionally, we performed qRT-PCR in breast cancer cell lines to verify the findings in integrated analysis. Results: After searching Gene Expression Omnibus database (GEO), two microarray studies were obtained according to the inclusion criteria. The integrated analysis was conducted, including 30 samples of TNBC and 77 samples of non-TNBC. 556 genes were found to be consistently differentially expressed (344 up-regulated genes and 212 down-regulated genes in TNBC). Functional annotation for these differentially expressed genes (DEGs) showed that the most significantly enriched Gene Ontology (GO) term for molecular functions was protein binding (GO: 0005515, P = 6.09E-21), while that for biological processes was signal transduction (GO: 0007165, P = 9.46E-08), and that for cellular component was cytoplasm (GO: 0005737, P = 2.09E-21). The most significant pathway was Pathways in cancer (P = 6.54E-05) based on Kyoto Encyclopedia of Genes and Genomes (KEGG). DUSP1 (Degree = 21), MYEOV2 (Degree = 15) and UQCRQ (Degree = 14) were identified as the significant hub proteins in the protein-protein interaction (PPI) network. Five genes were selected to perform qRT-PCR in seven breast cancer cell lines, and qRT-PCR results showed that the expression pattern of selected genes in TNBC lines and non-TNBC lines was nearly consistent with that in the integrated analysis. Conclusion: This study may help to understand the pathogenesis of different breast cancer subtypes, contributing to the successful identification of therapeutic targets for TNBC. [ABSTRACT FROM AUTHOR]

Major histocompatibility complex (MHC) genes are the most polymorphic genes in vertebrates and encode molecules that play a crucial role in pathogen resistance. As a result of their diversity, they have received much attention in the fields of evolutionary and conservation biology. Here, we described the genetic variation of MHC class II B (MHCIIB) exon 2 in a wild population of Hume’s pheasant (Syrmaticus humiae), which has suffered a dramatic decline in population over the last three decades across its ranges in the face of heavy exploitation and habitat loss. Twenty-four distinct alleles were found in 73 S. humiae specimens. We found seven shared alleles among four geographical groups as well as six rare MHCIIB alleles. Most individuals displayed between one to five alleles, suggesting that there are at least three MHCIIB loci of the Hume’s pheasant. The dN ⁄ dS ratio at putative antigen-binding sites (ABS) was significantly greater than one, indicating balancing selection is acting on MHCIIB exon 2. Additionally, recombination and gene conversion contributed to generating MHCIIB diversity in the Hume’s pheasant. One to three recombination events and seventy-five significant gene conversion events were observed within the Hume’s pheasant MHCIIB loci. The phylogenetic tree and network analysis revealed that the Hume’s pheasant alleles do not cluster together, but are scattered through the tree or network indicating a trans-species evolutionary mode. These findings revealed the evolution of the Hume’s pheasant MHC after suffering extreme habitat fragmentation. [ABSTRACT FROM AUTHOR]

The black-spotted tokay and the red-spotted tokay are morphologically distinct and have largely allopatric distributions. The black-spotted tokay is characterized by a small body size and dark skin with sundry spots, while the red-spotted tokay has a relatively large body size and red spots. Based on morphological, karyotypic, genetic, and distribution differences, recent studies suggested their species status; however, their classifications remain controversial, and additional data such as ecological niches are necessary to establish firm hypotheses regarding their taxonomic status. We reconstructed their ecological niches models using climatic and geographic data. We then performed niche similarity tests (niche identity and background tests) and point-based analyses to explore whether ecological differentiation has occurred, and whether such differences are sufficient to explain the maintenance of their separate segments of environmental ranges. We found that both niche models of the black- and the red-spotted tokay had a good fit and a robust performance, as indicated by the high area under the curve ( AUC) values ('black' = 0.982, SD = ± 0.002, 'red' = 0.966 ± 0.02). Significant ecological differentiation across the entire geographic range was found, indicating that the involvement of ecological differentiation is important for species differentiation. Divergence along the environmental axes is highly associated with climatic conditions, with isothermality being important for the 'black' form, while temperature seasonality, precipitation of warmest quarter, and annual temperature range together being important for the 'red' form. These factors are likely important factors in niche differentiation between the two forms, which result in morphological replacement. Overall, beside morphological and genetic differentiation information, our results contribute to additional insights into taxonomic distinction and niche differentiation between the black- and the red-spotted tokay. [ABSTRACT FROM AUTHOR]

□ Fertilization plays an important role in soil fertility and rice yield. A long-term (1990–2006) field experiment was conducted at a typical subtropical hilly agriculture region in south central China. The objectives of this study are to investigate the effects of long-term fertilizations on i) accumulations and distributions of soil carbon (C), nitrogen (N) and phosphorus (P) in different sizes of water-stable aggregates (WSA); ii) dynamic change of soil C, N, and P over years; and iii) the rice yield. Application of organic materials (OM), especially in combination with chemical fertilizers, increased soil organic C (SOC), soil total N (NT), and soil total P (PT) concentrations in all sizes of WSA, and they more concentrated in WSA (>1 mm). However, single chemical fertilizers had little impact on SOC and NT. There were significantly positive correlations between SOC and NTin each size of WSA, (r≥0.977, n = 15,P< 0.01). It illuminated that application of organic materials benefited the conservation of soil nitrogen. However, application of organic materials had less impact on PT. Variation of C/P was larger than C/N. PTwas mainly affected by input of phosphorus. Data over years showed that SOC and NTcontinued to rise, whereas PTtended to decline irrespective of treatments. Furthermore, economical chemical fertilizers application combining with green manure and half straw returning (NPKe+ SR) can produce approximate effects on PTas conventional dose of chemical fertilizers (NPKc). Average rice yield of NPKcfor 17 years was 9399 kg ha−1. Compared with NPKc, OM decreased yield by only 4.4%, which indicated that traditional organic fertilization could obtain relatively high yield. NPKe+ SR obtained approximate yield as NPKcdid, which showed that green manure and half amount of straw effectually reduced chemical fertilizers by about one third. [ABSTRACT FROM AUTHOR]

Aim. The complete mitochondrial genome sequence of a newly discovered vole, Proedromys liangshanensis (Rodentia: Cricetidae: Arvicolinae), was determined. Results. The mitogenome of P. liangshanensis is 16,296  bp in length. As with most other mammals, it contains the same gene order and an identical number of genes or regions, including 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and one putative control region. The A++T content of the control region is 58.2%, the lowest proportion detected so far in Myomorpha. To confirm the phylogenetic position of P. liangshanensis, we carried out phylogenetic analyses based on complete mitochondrial genomic data using Bayesian, maximum parsimony, and maximum likelihood methods. Conclusion. All results revealed that P. liangshanenis is sister to Microtus. Although the results do not bear light on the validity of the genus Proedromys, based on the morphological characters, we suggest that Proedromys is an independent genus of equal rank to the genus Microtus. [ABSTRACT FROM AUTHOR]

A vole species, Proedromys liangshanensis Liu, Sun, Zeng and Zhao, 2007 (Rodentia: Arvicolinae), which is endemic to the Liangshan Mountains, Sichuan, China, was reported in 2007. The species is restricted to a narrow range because of its weak dispersal ability and habitat specificity, and only lives in fir and spruce forests with abundant grass. In this study, the genetic diversity and phylogeographical structure of 42 P. liangshanensis were examined based on analysis of a partial mitochondrial DNA control region. The results revealed low genetic diversity in the P. liangshanensis populations. Twelve nucleotide haplotypes were defined in the 42 collected individuals, and were divided into two major phylogenetic clades, which were obviously related to geographic pattern. Estimates of gene flow (Nm = 0.09) also revealed a genetic divergence between the Jinyang and Meigu populations. Species-specific traits, habitat specificity and geographic events contributed to the genetic divergence of the new vole. [ABSTRACT FROM AUTHOR]

Swine finishing produces a large contribution of manure to nearby surface water and ground water in Jiulong upstream watershed, south Fujian Province, China. It is urgent to find a comprehensive, cost-effective way, not only to solve the manure pollution problem, but also to utilize the nutrients of manure for local farming. Gekou, a small hoggery village in the dense swine-farming core area of the upstream watershed, was selected as the study area. Mixed integer programming models based on uncertain interval numbers were employed to generate optimal whole farm plans for specialized hoggery. Emphasis was placed on discerning the most profitable ways to formulate hog feed rations, manure collection, manure storage, and manure application to achieve the maximum profit and the minimum pollution emission. Analyses were concentrated on ammonia emissions, excess total nitrogen and excess total phosphorus from manure operations, not only as nutrients applied to croplands, but also as pollutants to be minimized in compliance with environmental criteria. Three different hog feed rations, two alternative ways of collecting hog manure, and two alternative storage methods were combined with five alternatives for field application of manure to provide different combinations of hog feeding and manure-handling to be evaluated in the mixed integer programming models. Results indicated tradeoffs between economic goals and environmental goals, the latter being achieved only at some expense to farm net returns. The kaleyard and fishpond are recommended alternatives for field application of manure compared to three other alternatives. [ABSTRACT FROM AUTHOR]

The MDM2-p53 pathway has a prominent oncogenic function in the pathogenesis of various cancers. Nutlin-3, a small-molecule antagonist of MDM2-p53 interaction, inhibits proliferation in cancer cells with wild-type p53. Herein, we evaluate the expression of MDM2, both the full length and a splicing variant MDM2-A, and the sensitivity of Nutlin-3 in different cancer cell lines. Included are seven cell lines with wild-type p53 (four mesothelioma, one breast cancer, one chondrosarcoma, and one leiomyosarcoma), two liposarcoma cell lines harboring MDM2 amplification and wild-type p53, and one mesothelioma cell line harboring a p53 point mutation. Nutlin-3 treatment increased expression of cyclin D1, MDM2, and p53 in cell lines with wild-type p53. Additive effects were observed in cells containing wild-type p53 through coordinated attack on MDM2-p53 binding and cyclin D1 by lentivirual shRNA knockdown or small molecule inhibition, as demonstrated by immunoblots and cell viability analyses. Further results demonstrate that MDM2 binds to cyclin D1, and that an increase in cyclin D1 expression after Nutlin-3 treatment is correlated with expression and ubiquitin E3-ligase activity of MDM2. MDM2 and p53 knockdown experiments demonstrated inhibition of cyclin D1 by MDM2 but not p53. These results indicate that combination inhibition of cyclin D1 and MDM2-p53 binding warrants clinical evaluation as a novel therapeutic strategy in cancer cells harboring wild-type p53.

The first complete mitochondrial genome of the large white-bellied rat ( Niviventer excelsior) was sequenced and annotated in order to provide a source of phylogenetic characters including an assessment of gene order arrangement. The mitochondrial genome sequence (16,298 bp) of N. excelsior contains 13 protein-coding, 2 rRNA and 22 tRNA genes as well as 1 typical control region (D-Loop), which are shared by other mammalian mitochondrial genomes. The codon usage also followed the typical vertebrate pattern except for an unusual CCA stop codon, which was termination codon of Cyt b. The 16 nucleotide insertion was detected in the central conserved domain of d-Loop. The 12 heavy-strand encoded protein-coding genes were used to reconstruct the phylogenetic relationships by the Bayesian inference and maximum likelihood methods. Topologies revealed that N. excelsior clustered together with the species in genus Rattus, indicating their closer phylogenetic relationship. [ABSTRACT FROM AUTHOR]

Autophagy and glycolysis are highly conserved biological processes involved in both physiological and pathological cellular programs, but the interplay between these processes is poorly understood. Here, we show that the glycolytic enzyme lactate dehydrogenase A (LDHA) is activated upon UNC-51-like kinase 1 (ULK1) activation under nutrient deprivation. Specifically, ULK1 directly interacts with LDHA, phosphorylates serine-196 when nutrients are scarce and promotes lactate production. Lactate connects autophagy and glycolysis through Vps34 lactylation (at lysine-356 and lysine-781), which is mediated by the acyltransferase KAT5/TIP60. Vps34 lactylation enhances the association of Vps34 with Beclin1, Atg14L, and UVRAG, and then increases Vps34 lipid kinase activity. Vps34 lactylation promotes autophagic flux and endolysosomal trafficking. Vps34 lactylation in skeletal muscle during intense exercise maintains muscle cell homeostasis and correlates with cancer progress by inducing cell autophagy. Together, our findings describe autophagy regulation mechanism and then integrate cell autophagy and glycolysis.

Rewiring of redox metabolism has a profound impact on tumor development, but how the cellular heterogeneity of redox balance affects leukemogenesis remains unknown. To precisely characterize the dynamic change in redox metabolism in vivo, we developed a bright genetically encoded biosensor for H 2 O 2 (named HyPerion) and tracked the redox state of leukemic cells in situ in a transgenic sensor mouse. A H 2 O 2 -low (HyPerion-low) subset of acute myeloid leukemia (AML) cells was enriched with leukemia-initiating cells, which were endowed with high colony-forming ability, potent drug resistance, endosteal rather than vascular localization, and short survival. Significantly high expression of malic enzymes, including ME1/3, accounted for nicotinamide adenine dinucleotide phosphate (NADPH) production and the subsequent low abundance of H 2 O 2 . Deletion of malic enzymes decreased the population size of leukemia-initiating cells and impaired their leukemogenic capacity and drug resistance. In summary, by establishing an in vivo redox monitoring tool at single-cell resolution, this work reveals a critical role of redox metabolism in leukemogenesis and a potential therapeutic target.

Background: Osteosarcoma is a malignant bone tumor that typically occurs in adolescents or children under 20 years of age. Developing efficient clinical prognostic markers is crucial for improving the treatment of osteosarcoma patients., Methods: Three datasets related to osteosarcoma were acquired from the Gene Expression Omnibus (GEO) database. A gene signature model was established using the Limma package in the R software, univariate and multivariate survival analyses, and least absolute shrinkage and selection operator (LASSO) algorithms. The gene signature was then verified using external datasets., Results: From the GEO database, 242 differentially expressed genes were identified. A total of 590 unique genes, including 380 genes from the human protein interaction network, were found to be related to biological processes such as bone development and bone cell development. Univariate Cox survival analyses revealed 43 genes that were associated with the prognosis of osteosarcoma patients. A seven-gene signature [retinitis pigmentosa 2 ( RP2 ), polyhydroxybutyrate ( PHB ), myosin VI ( MYO6 ), mutL homolog 1 ( MLH1 ), Casein kinase 2 beta ( CSNK2B ), ribosomal protein L37A ( RPL37A ), and CCAAT/enhancer binding protein alpha ( CEBPA )] was developed using LASSO regression analysis and multivariate regression analysis. This gene signature could stratify the prognostic risk of sample cases in the training set, the test set, and the external verification set (P<0.01). The area under the receiver operating characteristic curve for the 5-year survival was higher than 0.72 in both the training and verification groups., Conclusions: In this study, a seven-gene signature was developed that is highly efficient at predicting the prognosis of patients with osteosarcoma, and therefore, this signature may be a crucial guide in the treatment of these patients., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://atm.amegroups.com/article/view/10.21037/atm-21-6276/ciof). The authors have no conflicts of interest to declare., (2022 Annals of Translational Medicine. All rights reserved.)

A new species of the frog genus Rana sensu lato from Wuyi Mountain, Fujian Province, China is described. Molecular phylogenetic analyses clustered the new species into the R.johnsi group and indicated that it was genetically divergent from its closely related species. The new species could be distinguished from its congeners by a combination of the following characters: body size medium, SVL 41.4-45.6 mm (42.9 ± 1.9 mm, n = 4) in adult males and 47.6-50.3 mm (n = 2) in adult females; adult male with a pair of internal subgular vocal sacs; lateroventral grooves present on tip of toes; webbing on fourth toes reaching the tip of toe; transverse skin ridges distinctly present on the dorsal surface of thigh and tibia, the number large (mean 26.5 ± 2.7, range 22-29, n = 6); breeding males possess creamy white nuptial pad with tiny velvety spines on the dorsal surface of the first finger, divided into three parts., (Yanqing Wu, Shengchao Shi, Huiguang Zhang, Weicai Chen, Bin Cai, Van Chung Hoang, Jun Wu, Bin Wang.)

We described a new species of the genus Leptobrachella from Southern Guangxi, China, based on morphological characteristics, molecular analyses and bioacoustics. This new species, Leptobrachella shiwandashanensis sp. nov., occurs in sympatry with Leptobrachella shangsiensis. However, phylogenetic analyses based on the mitochondrial 16S gene fragment revealed that the new species was clustered in a monophyletic group and a considerable genetic divergence existed between the new species and L. shangsiensis (p-distance: 7.9%) as well as its congeners (minimum p-distance: 7.0%). The new species also differed from its congeners by a combination of the following characteristics: (1) small size (SVL 26.829.7 mm in males; 33.735.9 mm in females); (2) pale brown dorsal surfaces, with a brown inverse-triangle-shaped marking between the eyes; (3) creamy white ventral surface with brown spots on the lateral margin, with a near immaculate creamy white throat and chest; (4) without webbing and lateral fringes on toes; (5) flanks with irregular black spots; (6) tibia-tarsal articulation reaching the posterior of the eye in males but reaching the shoulder in females; (7) heels that do not meet when the thighs are appressed at right angles to the body; (8) bicolored iris, with the upper half brownish-red, fading to silver in the lower half; and (9) a call consisting of a single note and a dominant frequency of 5.35.7 kHz (recorded at 23C).

This study describes a new member of the genus Leptobrachella, Leptobrachella shangsiensis sp. nov., based on morphological, acoustic and molecular analyses from southern Guangxi, China. L. shangsiensis sp. nov. differs from its congeners by a combination of the following characters: (1) SVL 24.9-29.4 mm in 11 males and 30.8-35.9 mm in 5 females, (2) black supratympanic fold with dark reddish rim, (3) ventrolateral glands distinct, forming a broken line, (4) ventral surface yellowish creamy-white with marble texture, (5) iris copper in the upper and silver in the lower fifth and (6) toe webbing basal and narrow lateral fringes. The male advertisement call of L. shangsiensis sp. nov. consists of one note per call, lacking an introductory note, with a mean dominant frequency of 5.5-6.5 kHz at the ambient temperature of 21.5ºC. Uncorrected sequence divergences between L. shangsiensis sp. nov. and all homologous DNA sequences available for the 16S rRNA gene were found to be > 5.9%. L. shangsiensis sp. nov. occurs in an evergreen forest in Shiwandashan National Nature Reserve located at an elevation of 450-550 m.

Rhacophorus yaoshanensis Liu Hu, 1962 and Theloderma kwangsiensis Liu Hu, 1962 were described by Liu Hu (1962) based on two specimens and one specimen, respectively, from the Dayaoshan Ranges, Guangxi, China. Since these two species were described, no additional specimens have been collected from their type localities, presenting an issue for phylogenetic studies of the genera. Five decades later, we have rediscovered R. yaoshanensis and T. kwangsiensis from their type localities. In this paper, we re-describe the two species and conduct a preliminary assessment of their phylogenetic relationships using two mitochondrial DNA genes (12S and 16S rRNA). The results indicate with high support that R. yaoshanensis is closely related to Rhacophorus pinglongensis. Theloderma kwangsiensis is nested within Theloderma corticale, with only 0.0-0.6% pairwise divergence, a level typical of intraspecific variation. Based on both molecular and morphological analyses, we further confirm that T. kwangsiensis is a synonym of T. corticale. Shiwandashan National Nature Reserve, Guangxi Province, China, is a new record for T. corticale.

Oncogenic KIT or PDGFRA receptor tyrosine kinase (RTK) mutations are compelling therapeutic targets in gastrointestinal stromal tumors (GIST), and treatment with the KIT/PDGFRA inhibitor imatinib is the standard of care for patients with metastatic GIST. Most GISTs eventually acquire imatinib resistance due to secondary mutations in the KIT kinase domain, but it is unclear whether these genomic resistance mechanisms require other cellular adaptations to create a clinically meaningful imatinib-resistant state. Using phospho-RTK and immunoblot assays, we demonstrate activation of KIT and insulin receptor (IR) in imatinib-resistant GIST cell lines (GIST430 and GIST48) and biopsies with acquisition of KIT secondary mutations, but not in imatinib-sensitive GIST cells (GIST882 and GIST-T1). Treatment with linsitinib, a specific IR inhibitor, inhibited IR and downstream intermediates AKT, MAPK, and S6 in GIST430 and GIST48, but not in GIST882, exerting minimal effect on KIT phosphorylation in these cell lines. Additive effects showing increased apoptosis, antiproliferative effects, cell-cycle arrest, and decreased pAKT and pS6 expression, tumor growth, migration, and invasiveness were observed in imatinib-resistant GIST cells with IR activation after coordinated inhibition of IR and KIT by linsitinib (or IR shRNA) and imatinib, respectively, compared with either intervention alone. IGF2 overexpression was responsible for IR activation in imatinib-resistant GIST cells, whereas IR activation did not result from IR amplification, IR mutation, or KIT phosphorylation. Our findings suggest that combinatorial inhibition of IR and KIT warrants clinical evaluation as a novel therapeutic strategy in imatinib-resistant GISTs. Cancer Res; 77(18); 5107-17. ©2017 AACR ., (©2017 American Association for Cancer Research.)

The expression and functions of microRNA (miR)-411 have been investigated in several types of cancer. However, until now, miR-411 in human breast cancer has not been examined. The present study investigated the expression, biological functions and molecular mechanisms of miR‑411 in human breast cancer, discussing whether it offers potential as a therapeutic biomarker for breast cancer in the future. The expression levels of miR‑411 in human breast cancer tissues and cells were measured using reverse transcription‑quantitative polymerase chain reaction analysis. Following transfection with miR‑411 mimics, an MTT assay, cell migration and invasion assay, western blot analysis and luciferase assay were performed in human breast cancer cell lines. According to the results, it was found that miR‑411 was significantly downregulated in breast cancer, and associated with lymph node metastasis and histological grade. Additionally, it was observed that miR‑411 suppressed cell growth, migration and invasion in the breast cancer cells. The present study also provided the first evidence, to the best of our knowledge, that miR‑411 was likely to directly target specificity protein 1 in breast cancer. These findings indicated that miR‑411 may be used a therapeutic biomarker for the treatment of breast cancer in the future.