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About 80% of hepatocellular carcinoma (HCC) patients are in advanced stages and ineligible for curative surgery. Palliative treatments just maintained limited survival, thus an effective downstaging therapy is badly needed. Here we report an initially unresectable patient who underwent radical hepatectomy after successful downstaging with selective internal radiation therapy (SIRT). A 34‐year‐old man was diagnosed with China Liver Cancer Staging (CNLC) IIIa HCC. Due to insufficient future liver remnant and vascular involvement, the patient was suggested to be unresectable. SIRT with yttrium‐90 resin microspheres was given. At three months post‐SIRT, a complete response was achieved. The tumor was downstaged to CNLC Ia stage. The patient underwent anatomical hepatectomy 5 months after SIRT. Histopathological examination of the resected specimen showed 4% viable tumor cells inside a necrotic mass. To our knowledge, this is the first case who underwent SIRT with yttrium‐90 resin microspheres in China mainland. The success of the downstaging in this case renders a possible cure to be achieved in an initially unresectable patient. In addition, the nearly complete tumor necrosis in the resected specimen indicates a good prognosis post‐surgery. This is the first case who underwent SIRT with yttrium‐90 resin microspheres in China mainland. SIRT followed by anatomical hepatectomy is a potentially curative strategy for unresectable HCC, which deserves a confirmative trial in the future. [ABSTRACT FROM AUTHOR]

In the field of image classification, the Convolutional Neural Networks (CNNs) are effective. Most of the work focuses on improving and innovating CNN's network structure. However, using labeled data more effectively for training has also been an essential part of CNN's research. Combining image disturbance and consistency regularization theory, this paper proposes a model training method (PairTraining) that takes image pairs as input and dynamically modify the training difficulty according to the accuracy of the model in the training set. According to the accuracy of the model in the training set, the training process will be divided into three stages: the qualitative stage, the fine learning stage and the strengthening learning stage. Contrastive learning images are formed using a progressively enhanced image disturbance strategy at different training stages. The input image and contrast learning image are combined into image pairs for model training. The experiments are tested on four public datasets using eleven CNN models. These models have different degrees of improvement in accuracy on the four datasets. PairTraining can adapt to a variety of CNN models for image classification training. This method can better improve the effectiveness of training and improve the degree of generalization of classification models after training. The classification model obtained by PairTraining has better performance in practical application. [ABSTRACT FROM AUTHOR]

Keloids are benign dermal tumors that arise from abnormal wound healing processes following skin lesions. Surgical excision followed by radiotherapy plays an important role in the treatment of keloids. Nevertheless, radioresistance remains a serious impediment to treatment efficacy. Investigation of the molecular response of keloids to radiation may contribute to radiosensitizing strategies. Primary keloid fibroblasts from human keloids were isolated and irradiated with X-ray. The expression profiles of messenger RNA (mRNA) in nonradiated and irradiated primary keloid fibroblasts were measured by mRNA sequencing analysis. Then, we identified common motifs and corresponding transcription factors of dysregulated mRNAs by using bioinformatic analysis of the proximal promoters. Whereafter, GO and KEGG were used to analyze the functional enrichment of the differentially expressed genes. We found that radiation not only suppressed proliferation but also increased cell senescence of primary keloid fibroblasts. There were 184 mRNAs and 204 mRNAs that showed significant changes in 4 and 8 Gy irradiated primary keloid fibroblasts, respectively. Among them, 8 upregulated and 30 downregulated mRNAs showed consistent alterations in 4 and 8 Gy irradiated primary keloid fibroblasts. More importantly, the xForkhead box O1 (FOXO1) signaling pathway was involved in the irradiation response. Pretreatment with the FOXO1 signaling inhibitor AS1842856 significantly promoted LDH release, apoptosis and senescence of primary keloid fibroblasts following irradiation. Our findings illustrated the molecular changes in human keloid fibroblasts in response to radiation, and FOXO1 pathway inhibition is expected to provide a novel strategy for the radiosensitization of keloids. [ABSTRACT FROM AUTHOR]

Radon is a naturally occurring radioactive gas and considered as a serious carcinogen to humans. Continuous radioactive decay of this gas emits high-energy alpha particles. Long-term radon exposure induces oxidative stress and inflammatory response, which results in chronic lung diseases. However, biological effects after radon exposure in other organs have been rarely reported. As the outermost organ of the human body, the skin suffers from environmental damage to agents such as air pollution. Epidemiological studies indicated that areas with high level of radon had a high incidence of skin cancer. However, whether radon exposure induces skin damage has not been reported yet. In this study, we established a radon-exposed mouse model and found that radon exposure affected the structure of skin tissues, which was manifested by inflammatory cell infiltration and skin atrophy. Using proteomic approach, we found 45 preferentially expressed proteins in 60 Working Level Months (WLM) group and 314 preferentially expressed proteins in 120 WLM group from radon-exposed skin tissues. Through microRNA (miRNA) sequencing profiling analysis, 57 dysregulated miRNAs were screened between the control and radon-treated mouse skin. By integrating the dysregulated proteins and miRNAs, radon-induced fatty acid synthase (FASN) was investigated in greater detail. Results showed that FASN was regulated by miR-206-3p and miR-378a-3p and involved in the pathogenesis of radon-induced skin damage. Overexpression of FASN inhibited the proliferation, and induced in WS1 cells. Our present findings illustrate the molecular change during radon-induced skin damage and the potential role of FASN during this process. [ABSTRACT FROM AUTHOR]

BRAF mutation is a representative oncogenic mutation, with a frequency of 60% in papillary thyroid carcinoma (PTC), but the reasons for the poor prognosis and more aggressive course of BRAF-mutated PTC are controversial. Tumor immune microenvironment (TIME) is an essential factor permitting the development and progression of malignancy, but whether TIME participates in the prognosis of BRAF-mutated PTC has not yet been reported. The primary goal of the present study was to provide a comprehensive TIME-related prognostic model to increase the predictive accuracy of progression-free survival (PFS) in patients with BRAF-mutated PTC. In this study, we analyzed the mRNA-seq data and corresponding clinical data of PTC patients obtained from the TCGA database. By calculating the TIME scores (immune score, stromal score and ESTIMATE score), the BRAF mutation group (n=237) was dichotomized into the high- and low-score groups. By functional analysis of differentially expressed genes (DEGs) in different high/low score groups, we identified 2 key TIME-related genes, HTR3A and NIPAL4 , which affected PFS in BRAF-mutated PTC. A risk scoring system was developed by multivariate Cox analysis based on the abovementioned 2 TIME-related genes. Then, the BRAF-mutated cohort was divided into the high- and low-risk groups using the median risk score as a cutoff. A high risk score correlated positively with a higher HTR3A/NIPAL4 expression level but negatively with PFS in BRAF-mutated PTC. Ultimately, a nomogram was constructed by combining risk score with clinical parameter (Tumor stage), and the areas under the ROC curve (AUCs) of the nomogram for predicting 1-, 3- and 5-year PFS were then calculated and found to be 0.694, 0.707 and 0.738, respectively, indicating the improved accuracy and clinical utility of the nomogram versus the risk score model in the BRAF-mutated PTC cohort. Moreover, we determined the associations between prognostic genes or risk score and immune cell infiltration by two-way ANOVA. In the high-risk score, high HTR3A expression, and high NIPAL4 expression groups, higher infiltration of immune cells was found. Collectively, these findings confirm that the nomogram is effective in predicting the outcome of BRAF-mutated PTC and will add a spatial dimension to the developing risk stratification system. [ABSTRACT FROM AUTHOR]

Papillary thyroid cancer (PTC) accounts for about 90% of thyroid cancer. There are approximately 20%–30% of PTC patients showing disease persistence/recurrence and resistance to radioactive iodine (RAI) treatment. For these PTC patients with RAI refractoriness, the prognosis is poor. In this study, we aimed to establish a comprehensive prognostic model covering multiple signatures to increase the predictive accuracy for progression-free survival (PFS) of PTC patients with RAI treatment. The expression profiles of mRNAs and miRNAs as well as the clinical information of PTC patients were extracted from TCGA and GEO databases. A series of bioinformatics methods were successfully applied to filtrate a two-RNA model (IPCEF1 and hsa-mir-486-5p) associated with the prognosis of RAI-therapy. Finally, the RNA-based risk score was calculated based on the Cox coefficient of the individual RNA, which achieved good performances by the time-dependent receiver operating characteristic (tROC) curve and PFS analyses. Furthermore, the predictive power of the nomogram, integrated with the risk score and clinical parameters (age at diagnosis and tumor stage), was assessed by tROC curves. Collectively, our study demonstrated high precision in predicting the RAI response of PTC patients. [ABSTRACT FROM AUTHOR]

Along with the development of sonar technology, the detection accuracy and stability of sonar have been improved. A large amount of seabed sediment information can be obtained through sonar detection. However, this information is often accompanied by noise interference, resulting in poor quality of the generated images. Moreover, sonar images are different from conventional images. There are single-channel images. The model needs to classify the images according to the texture features in the images. Coupled with the scarcity of sonar data, this makes it difficult to accurately classify the seabed sediment. According to the characteristics of sonar images, we propose the ShuffleNet-DSE which is a classification model based on deep learning. The ShuffleNet-DSE network is based on ShuffleNet-V2, while ensuring the lightweight of the network, it incorporates feature dense connection and Squeeze-and-Excitation (SE) structure channel self-attention. And combined with the sonar image's characteristics, the partial activation function of the model is changed to the Swish. The experimental results show that compared with the traditional machine learning classification method, ShuffleNet-DSE has greatly improved the classification accuracy and the computational cost. Compared with excellent neural network models such as AlexNet, MobileNet-V3, GoogLeNet and ResNet, it is more suitable for sonar image processing. [ABSTRACT FROM AUTHOR]

Radiation-induced esophageal injury (RIEI) is a major dose-limiting complication of radiotherapy, especially for esophageal and thoracic cancers. RIEI is a multi-factorial and multi-step process, which is regulated by a complex network of DNA, RNA, protein and metabolite. However, it is unclear which esophageal metabolites are altered by ionizing radiation and how these changes affect RIEI progression. In this work, we established a rat model of RIEI with 0–40 Gy X-ray irradiation. Esophageal irradiation using ≥25 Gy induced significant changes to rats, such as body weight, food intake, water intake and esophageal structure. The metabolic changes and related pathways of rat esophageal metabolites were investigated by liquid chromatography-mass spectrometry (LC-MS). One hundred eighty metabolites showed an up-regulation in a dose-dependent manner (35 Gy ≥ 25 Gy > controls), and 199 metabolites were downregulated with increasing radiation dose (35 Gy ≤ 25 Gy < controls). The KEGG analysis showed that ionizing radiation seriously disrupted multiple metabolic pathways, and arachidonic acid metabolism was the most significantly enriched pathway. 20 metabolites were dysregulated in arachidonic acid metabolism, including up-regulation of five prostaglandins (PGA2, PGJ2, PGD2, PGH2, and PGI2) in 25 or 35 Gy groups. Cyclooxygenase-2 (COX-2), the key enzyme in catalyzing the biosynthesis of prostaglandins from arachidonic acid, was highly expressed in the esophagus of irradiated rats. Additionally, receiver operating characteristic (ROC) curve analysis revealed that PGJ2 may serve as a promising tissue biomarker for RIEI diagnosis. Taken together, these findings indicate that ionizing radiation induces esophageal metabolic alterations, which advance our understanding of the pathophysiology of RIEI from the perspective of metabolism. [ABSTRACT FROM AUTHOR]

V-Raf murine sarcoma viral oncogene homolog B1 (BRAF) encodes a serine-threonine kinase. The V600E point mutation in the BRAF gene is the most common mutation, predominantly occurring in melanoma, and colorectal, thyroid and non-small cell lung cancer. Particularly in the context of thyroid cancer research, it is routinely employed as a molecular biomarker to assist in diagnosing and predicting the prognosis of papillary thyroid cancer (PTC), and to formulate targeted therapeutic strategies. Currently, several methods are utilized in clinical settings to detect BRAF V600E mutations in patients with PTC. However, the sensitivity and specificity of various detection techniques vary significantly, resulting in diverse detection outcomes. The present review highlights the advantages and disadvantages of the methods currently employed in medical practice, with the aim of guiding clinicians and researchers in selecting the most suitable detection approach for its high sensitivity, reproducibility and potential to develop targeted therapeutic regimens for patients with BRAF gene mutation-associated PTC. [ABSTRACT FROM AUTHOR]

Neurovascular dysfunction is a primary or secondary cause in the pathogenesis of several cerebrovascular and neurodegenerative disorders, including stroke. Therefore, the overall protection of the neurovascular unit (NVU) is a promising therapeutic strategy for various neurovascular diseases. However, the complexity of the NVU limits the study of the pathological mechanisms of neurovascular dysfunction. Reconstituting the in vitro NVU is important for the pathological study and drug screening of neurovascular diseases. In this study, we generated a spontaneously assembled three‐dimensional NVU (3D NVU) by employing the primary neural stem cells and brain microvascular endothelial cells in a Matrigel extracellular matrix platform. This novel model exhibits the fundamental structures and features of the NVU, including neurons, astrocytes, oligodendrocytes, vascular‐like structures, and blood–brain barrier‐like characteristics. Additionally, under oxygen‐glucose deprivation, the 3D NVU exhibits the neurovascular‐ or oxidative stress‐related pathological characteristics of cerebral ischemia and the injuries can be mitigated, respectively, by supplementing with the vascular endothelial growth factor or edaravone, which demonstrated that the availability of 3D NVU in ischemic stroke modeling. Finally, the 3D NVU promoted the angiogenesis and neurogenesis in the brain of cerebral ischemia rats. We expect that the proposed in vitro 3D NVU model will be widely used to investigate the relationships between angiogenesis and neurogenesis and to study the pathology and pharmacology of neurovascular diseases. [ABSTRACT FROM AUTHOR]

Graves' ophthalmopathy (GO) has become one of the most common orbital diseases. Although some evidences announced the potential mechanism of pathological changes in extraocular muscle and orbital adipose tissue, little is known about that in lacrimal enlargement of GO patients. Thus, gene expression profiles of lacrimal gland derived from GO patients and normal controls were investigated using the microarray datasets of GSE105149 and GSE58331. The raw data and annotation files of GSE105149 and GSE58331 were downloaded from Gene Expression Omnibus (GEO) database. Bioinformatics including differentially expressed genes (DEGs), Gene Ontology, Kyoto Encyclopedia of Gene and Genome (KEGG) pathway, protein-protein interaction (PPI) network construction, hub gene identification, and gene set variation analysis (GSVA) were successively performed. A total of 173 overlapping DEGs in GSE105149 and GSE58331 were screened out, including 20 up-regulated and 153 down-regulated genes. Gene Ontology, KEGG and GSVA analyses of these DEGs showed that the most significant mechanism was closely associated with endoplasmic reticulum (ER). Moreover, we identified 40 module genes and 13 hub genes which were also enriched in the ER-associated terms and pathways. Among the hub genes, five genes including HSP90AA1 , HSP90B1 , DNAJC10 , HSPA5 , and CANX may be involved in the dysfunction of protein processing in ER. Taken together, our observations revealed a dysregulated gene network which is essential for protein processing in ER in GO patients. These findings provided a potential mechanism in the progression of lacrimal enlargement in GO patients, as a new insight into GO pathogenesis. [ABSTRACT FROM AUTHOR]

Serine protease inhibitors (SPIs, serpins) are a protein superfamily involved in almost all physiological processes in all organisms. In this study, a novel serpin was identified from Apostichopus japonicus (Ajserpin) by using high-throughput sequencing and RACE approaches. The full-length cDNA of Ajserpin was 1893 bp with a 5′-untranslated region (UTR) of 130 bp, a 3′-UTR of 587 bp, and an open reading frame of 1176 bp encoding a polypeptide of 391 amino acids with a deduced molecular weight of 43.8 kDa. Ajserpin shares the standard structure of SPI, including three β-sheets and eight α-helices. The deduced amino acid sequences of Ajserpin had no nuclear location signal and signal peptide structure. The phylogenetic tree and immunofluorescence showed that Ajserpin belonged to the clade B subfamily and was mainly located in the cytoplasm and nucleus. Sequence comparison and protein inhibition experiments showed that the active site (P1–P1' site) of Ajserpin was Arginine and Serine, which displayed inhibitory activity toward trypsin in a dose-dependent manner. Tissue distribution analysis showed that Ajserpin transcripts were constitutively expressed in all examined tissues with the peak in the body wall. Ajserpin mRNA transcripts could be induced in Vibrio splendidus -challenged sea cucumber or lipopolysaccharide-exposed coelomocytes. Furthermore, Ajserpin knockdown by small interfering RNAs could inhibit coelomocytes apoptosis. All our results revealed that Ajserpin might serve as an immune regulator in sea cucumber. • Complete cDNA sequence of serine protease inhibitors was identified in Apostichopus japonicus. • Immunofluorescence showed Ajserpin was an intracellular protein belonging to the Clade B sub-family. • Ajserpin mRNA transcripts could be induced by Vibrio splendidus challenge or LPS exposure. • RAjserpin displayed inhibitory activity towards trypsin in a dose-dependent manner. • Ajserpin knockdown by siRNA could accelerate the coelomocytes apoptosis. [ABSTRACT FROM AUTHOR]

Ampullofolliculina Hadži, 1951 is a monotypic genus, the only species being A. lageniformis which was first discovered in estuarine habitats in the U.S. and has never been redescribed. In the present study, we investigated the taxonomy and phylogeny of A. lageniformis Hadži, 1951 based on analyses of a population collected from a brackish wetland in Ningbo, China. The main characteristics of this species are as follows: trophont about 450–700 μm long in vivo with two short, rounded peristomial lobes of equal size; lorica flask‐shaped, transparent and smooth with a short, wide neck at the base of which two transparent valves are asymmetrically inserted; cortex with brownish to reddish cortical granules and greenish pigment granules; about 80 somatic kineties evenly arranged; moniliform macronucleus with 4–8 ellipsoidal nodules; swarmer dark green, vermiform in shape, about 200–350 μm long in vivo, with about 60 adoral membranelles and 85 somatic kineties, no mouth nor paroral membrane. Phylogenetic analyses inferred from SSU rDNA sequences show that A. lageniformis is closely related to Folliculina and Eufolliculina which nest within a large clade that comprises five families, i.e. Stentoridae, Blepharismidae, Fabreidae, Maristentoridae, and Folliculinidae. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to assess the differences in infection rates between rituximab (RTX) and non‐RTX treatment in patients with rheumatoid arthritis (RA). Methods: A systematic review and meta‐analysis was conducted by searching databases of PubMed, MEDLINE, EMBASE, Web of Science and Cochrane Library through to June 2018. We included studies that compared RTX and non‐RTX treatment for patients with RA. Outcome measures were overall infections and serious infections between RTX and non‐RTX treatments. Results: A total of 11 articles, including 9502 patients (4595 with RTX treatment and 4907 with non‐RTX treatment) met our inclusion criteria. The results demonstrated that RTX‐related all infections and serious infections in RA patients were 43.8% and 4.4%, respectively. Pooled analysis showed no significant differences between RTX and non‐RTX treatment groups in overall infections rate (43.3% vs 44.9%; odds ratio [OR] = 0.87; 95% CI = 0.70‐1.08) and serious infections rate (4.1% vs 4.6%; OR = 1.05; 95% CI = 0.84‐1.31). Subgroup analysis also showed no significant differences in overall infections between RTX versus placebo (OR = 0.98, 95% CI = 0.71‐1.33); RTX versus tumor necrosis factor inhibitors (TNFi) (OR = 0.47, 95% CI = 0.30‐1.73); RTX plus methotrexate (MTX) versus placebo plus MTX (OR = 0.98, 95% CI = 0.77‐1.24), and in serious infections between RTX versus placebo (OR = 1.06, 95% CI = 0.36‐3.07); RTX versus TNFi (OR = 1.25, 95% CI = 0.96‐1.63); RTX plus MTX versus placebo plus MTX (OR = 0.69, 95% CI = 0.39‐1.20). Conclusion: In patients with RA, RTX treatment has no additional risks for infections over non‐RTX treatment. [ABSTRACT FROM AUTHOR]

Previous experiments showed that high concentrations of ethyl tertiary butyl ether (ETBE) exposure (500–5,000 ppm) significantly resulted in DNA damages in aldehyde dehydrogenase 2 (Aldh2) knockout (KO) mice. This study was aimed to verify the genotoxic effects in three genetic types, Aldh2 KO, heterogeneous (HT), and wild type (WT), of mice exposed to lower concentrations of ETBE (50–500 ppm) by inhalation. Histopathology assessments in the livers, measurements of genotoxic biomarkers in blood and livers, and urinary 8‐hydroxydeoxyguanosion (8‐OH‐dG) for the oxidative DNA damage of whole body were performed. Significant histopathological changes and DNA strand breaks both in hepatocytes and leukocytes were found in HT and KO male mice exposed to ≥200 ppm ETBE, but not in 50 ppm ETBE. 8‐OH‐dG levels either in liver or urine were higher in the HT and KO male mice exposed to ≥200 ppm ETBE. The pathological and genetic effects of ETBE were almost at the same extents for HT and KO mice. Thus, 50 ppm could be the no observed adverse effect level for ETBE in HT and KO male mice, which was far lower than the 500 ppm in WT mice. These results suggested that decrease and deficiency of ALDH2 activity would significantly increase the sensitivity to ETBE‐induced genotoxicity as well as hepatotoxic effects after exposure even to low concentrations of ETBE. Environ. Mol. Mutagen. 60: 145–153, 2019. © 2018 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

Abstract C-type lectins (CTLs) are important pattern recognition molecules that participate in bacterial binding and agglutination by specific recognition of carbohydrates from pathogens. In this study, a full-length cDNA of CTL was cloned from Sinonovacula constricta (designated ScCTL-2). ScCTL- 2 has a length of 981 bp, a 5′-untranslated region (UTR) of 47 bp, a short 3′-UTR of 37 bp, and an open reading frame (ORF) of 894 bp, which encodes a polypeptide of 298 amino acid residues. The deduced amino acid of ScCTL-2 possesses a conserved carbohydrate-recognition domain (CRD) similar to that of C31–E171. Spatial distribution analysis demonstrated that ScCTL-2 was constitutively expressed in all tested tissues, with dominant expression in foot and siphon and weak expression in hepatopancreas. The mRNA expression level of ScCTL-2 in gills and hepatopancreas was significantly upregulated at 6 and 12 h after challenge with the pathogen Vibrio parahaemolyticus. The recombinant ScCTL-2 showed specific binding and agglutinate capacities to all examined Gram-negative bacterial species, namely, Escherichia coli , Vibro anguillarum , and V. parahaemolyticus in a Ca2+-independent manner. However, these binding activities were not detected in Gram-positive Micrococcus luteus. Our results indicated that ScCTL-2 could be a novel pattern recognition receptor that can specifically recognize Gram-negative microorganisms in the innate immunity of S. constricta. Highlights • A novel Ca2+-independent C-type lectin was cloned from Sinonovacula constricta. • ScCTL-2 was constitutively expressed in all of the examined tissues with the highest magnitude in foot and siphon. • Vibrio splenfidus could significantly up-regulate the expressions of ScCTL-2 in hepatopancreas and gill tissues. • Recombinant ScCTL-2 can bind and agglutinate all tested G− bacteria, not G+ bacteria in a Ca2+-independent manner. [ABSTRACT FROM AUTHOR]

ABSTRACT Abnormal feedback of serum calcium to parathyroid hormone (PTH) secretion is the hallmark of primary hyperparathyroidism (PHPT). Although the molecular pathogenesis of parathyroid neoplasia in PHPT has been linked to abnormal expression of genes involved in cell growth (e.g., cyclin D1, retinoblastoma, and β-catenin), the molecular basis of abnormal calcium sensing by calcium-sensing receptor (CaSR) and PTH hypersecretion in PHPT are incompletely understood. Through gene expression profiling, we discovered that an orphan adhesion G protein-coupled receptor (GPCR), GPR64/ADGRG2, is expressed in human normal parathyroid glands and is overexpressed in parathyroid tumors from patients with PHPT. Using immunohistochemistry, Western blotting, and coimmunoprecipitation, we found that GPR64 is expressed on the cell surface of parathyroid cells, is overexpressed in parathyroid tumors, and physically interacts with the CaSR. By using reporter gene assay and GPCR second messenger readouts we identified Gαs, 3′,5′-cyclic adenosine monophosphate (cAMP), protein kinase A, and cAMP response element binding protein (CREB) as the signaling cascade downstream of GPR64. Furthermore, we found that an N-terminally truncated human GPR64 is constitutively active and a 15-amino acid-long peptide C-terminal to the GPCR proteolysis site (GPS) of GPR64 activates this receptor. Functional characterization of GPR64 demonstrated its ability to increase PTH release from human parathyroid cells at a range of calcium concentrations. We discovered that the truncated constitutively active, but not the full-length GPR64 physically interacts with CaSR and attenuates the CaSR-mediated intracellular Ca2+ signaling and cAMP suppression in HEK293 cells. Our results indicate that GPR64 may be a physiologic regulator of PTH release that is dysregulated in parathyroid tumors, and suggest a role for GPR64 in pathologic calcium sensing in PHPT. © 2016 American Society for Bone and Mineral Research. [ABSTRACT FROM AUTHOR]

Primary hyperparathyroidism ( PHPT) is a common endocrine neoplastic disorder caused by a failure of calcium sensing secondary to tumour development in one or more of the parathyroid glands. Parathyroid adenomas are comprised of distinct cellular subpopulations of variable clonal status that exhibit differing degrees of calcium responsiveness. To gain a clearer understanding of the relationship among cellular identity, tumour composition and clinical biochemistry in PHPT, we developed a novel single cell platform for quantitative evaluation of calcium sensing behaviour in freshly resected human parathyroid tumour cells. Live-cell intracellular calcium flux was visualized through Fluo-4- AM epifluorescence, followed by in situ immunofluorescence detection of the calcium sensing receptor ( CASR), a central component in the extracellular calcium signalling pathway. The reactivity of individual parathyroid tumour cells to extracellular calcium stimulus was highly variable, with discrete kinetic response patterns observed both between and among parathyroid tumour samples. CASR abundance was not an obligate determinant of calcium responsiveness. Calcium EC50 values from a series of parathyroid adenomas revealed that the tumours segregated into two distinct categories. One group manifested a mean EC50 of 2.40 mM (95% CI: 2.37-2.41), closely aligned to the established normal range. The second group was less responsive to calcium stimulus, with a mean EC50 of 3.61 mM (95% CI: 3.45-3.95). This binary distribution indicates the existence of a previously unappreciated biochemical sub-classification of PHPT tumours, possibly reflecting distinct etiological mechanisms. Recognition of quantitative differences in calcium sensing could have important implications for the clinical management of PHPT. [ABSTRACT FROM AUTHOR]

The skin is the largest outermost organ of the human body. It is vulnerable to various damages, such as ionizing radiation. Exploration of proliferation, senescence and radiosensitivity of skin cells contributes to the development of medical and cosmetic countermeasures against skin aging and toward injury protection. Human antigen R (HuR) is one of the most widely studied RNA-binding proteins and serves an important role in stabilization of mRNA and regulation of the expression of the target genes. To investigate the role of HuR in modulating proliferation, senescence and radiosensitivity of skin cells, the present study performed an in vitro study using lentivirus-mediated overexpression or silencing of HuR in human keratinocyte HaCaT cells and human skin fibroblast WS1 cells. The results indicated that overexpression of HuR promoted proliferation, whereas downregulation of HuR inhibited proliferation of HaCaT and WS1 cells. Overexpression of HuR reduced apoptosis and senescence in skin cells. RNA-Seq of skin cells with HuR overexpression or knockdown identified 77 mRNAs positively or negatively correlated with HuR expression levels. In addition, silencing of HuR induced a significant increase in radiogenic reactive oxygen species after irradiation. Overexpression of HuR increased radiotolerance of HaCaT and WS1 cells. RNA immunoprecipitation coupled with RNA-Seq identified 14 mRNAs interacting with HuR upon radiation exposure. Overall, the findings of the present study illustrated the key role of HuR in modulating proliferation, senescence and radiosensitivity of skin cells providing a new therapeutic strategy for cosmetic treatments and to combat skin injury. [ABSTRACT FROM AUTHOR]

AU-rich element RNA-binding factor 1 (AUF1) is a classical RNA-binding protein. AUF1 influences the process of development, apoptosis and tumorigenesis by interacting with adenylate-uridylate rich element-bearing mRNAs. Human skin is the largest organ of the body and acts as a protective barrier against pathogens and injuries. The aim of the present study was to explore the function and potential molecular pathways of AUF1 in human skin cells. AUF1 was overexpressed in human keratinocyte HaCaT cells and human skin fibroblast WS1 cells using adenoviruses and silenced using lentiviruses. AUF1 overexpression facilitated cell proliferation, whereas AUF1 knockdown induced the opposite effect. AUF1 reduced apoptosis but did not affect cell cycle progression. Forced AUF1 expression promoted the migration of human skin cells, as demonstrated by a scratch wound healing assay. Cell senescence was alleviated in AUF1-overexpressing skin cells, while AUF1 knockdown increased cell senescence. WS1 cells with AUF1 overexpression and silencing were used for RNA-sequencing and Kyoto Encyclopedia of Genes and Genomes-based pathway analysis to identify AUF1-affected mRNAs. A total of 18 mRNAs (eight mRNAs with positive associations and 10 mRNAs with negative associations) revealed consistent associations with both AUF1 overexpression and silencing. Enriched pathways associated with AUF1 expression included 'MAPK', 'cell adhesion molecules', 'proteasome', 'cellular senescence' and 'TGF-β signaling', indicating a complex regulatory network. Overall, the results of the present study revealed that AUF1 is involved in the proliferation, migration and senescence of skin cells in vitro and may be a potential target for cosmetic and disease treatment of skin. [ABSTRACT FROM AUTHOR]

ARC5 is a dynamin-related GTPase essential for the division of chloroplasts in plants. The arc5 mutant frequently exhibits enlarged, dumbbell-shaped chloroplasts, indicating a role for ARC5 in the constriction of the chloroplast division site. In a screen for chloroplast division mutants with a phenotype similar to arc5, two mutants, cpd25 and cpd45, were obtained. CPD45 was identified as being the same gene as FHY3, a key regulator of far-red light signaling recently shown to be involved in the regulation of ARC5. CPD25 was previously named FRS4 and is homologous to FHY3. We found that CPD25 is also required for the expression of ARC5, suggesting that its function is not redundant to that of FHY3. Moreover, cpd25 does not have the far-red light-sensing defect present in fhy3 and far1. Both FRS4/ CPD25 and FHY3/ CPD45 could bind to the FBS-like ' ACGCGC' motifs in the promoter region of ARC5, and the binding efficiency of FRS4/ CPD25 was much higher than that of FHY3/ CPD45. Unlike FHY3/ CPD45, FRS4/ CPD25 has no ARC5 activation activity. Our data suggest that FRS4/ CPD25 and FHY3/ CPD45 function as a heterodimer that cooperatively activates ARC5, that FRS4/ CPD25 plays the major role in promoter binding, and that FHY3/ CPD45 is largely responsible for the gene activation. This study not only provides insight into the mechanisms underlying the regulation of chloroplast division in higher plants, but also suggests a model that shows how members of a transcription factor family can evolve to have different DNA-binding and gene activation features. [ABSTRACT FROM AUTHOR]

Abstract: Interactions between all ten mature proteins of the potyviruses Soybean mosaic virus (Pinellia ternata isolate) and Shallot yellow stripe virus were investigated using yeast two-hybrid (Y2H) assays. Consistently strong self-interactions were found between the pairs of HC-Pro, VPg, NIa-Pro, NIb and CP in both viruses. Apart from the NIb, such interactions have been previously reported for some other potyviruses. The 6K1/NIa-Pro combination gave a consistently moderate to strong interaction in both directions for both viruses. This interaction occurred even when the 6K1 of SMV-P was truncated to eliminate the C-terminal motif that acts as a recognition site for cleavage by the NIa-Pro. Many other interactions occurred only in one direction or only for one of the two viruses. When taken together with other published reports, the data suggest that interactions detected by Y2H should be regarded as only preliminary indications. [Copyright &y& Elsevier]

• Two folliculinids are redescribed based on detailed morphological characters. • The life cycles of two folliculinids are described in detail for the first time. • SSU rDNA sequences of two folliculinid species are determined. • The taxonomic confusion and mistakes of M. producta are corrected and clarified. • Evolutionary relationships of Folliculinidae are revealed by phylogenetic analyses. Most ciliates of the heterotrich family Folliculinidae are flagship periphytic species. They are characterized by their transparent lorica, conspicuous peristomial lobes, and dimorphic life cycle. However, the understanding of their biodiversity and systematics is relatively poor. In the present study, we investigate the morphology and phylogenetic affinities of Ampullofolliculina lageniformis Hadži, 1951 and Metafolliculina producta (Wright, 1859) Dons, 1934, collected from subtropical coastal wetlands in China. An integrative approach combining investigations of their morphology, life cycle and ecology was used. Phylogenetic analyses based on small subunit ribosomal DNA (SSU rDNA) sequences showed that: all folliculinid genera form a monophyletic clade; the two genera with flexible peristomial lobes, Metafolliculina and Eufolliculina , are closely related; the other three genera with inflexible peristomial lobes, Ampullofolliculina , Diafolliculina and Folliculina, group together. [ABSTRACT FROM AUTHOR]

In this study, tea polyphenols (TPs) was first extracted from tea waste by superheated steam (SS) pretreated ultrasonic‐assisted hydrothermal extraction (UAH). The optimized strategy presented extracts with the extraction yield up to 21.19% with a significantly higher antioxidant ability, compared with the one without SS pretreatment. Further investigation proved that the SS suppressed the polyphenol oxidase activity of the TPs extract. The ability to scavenge the free radicals were compared in mouse liver mitochondria. Mitochondrial swelling, mitochondrial membrane potential (MMP), cardiolipin peroxidation, and respiratory chain complex (RCC) I–V activities were also evaluated as the index of the mitochondrial oxidative damage. The study supports evidence that the TPs extract exhibited significant protection against oxidative damage on mitochondrial. Furthermore, the effect of TPs on antioxidant ability in zebrafish embryo was evaluated. After TPs pretreatment for 1 day, zebrafish embryos showed a significantly higher survival rate as well as heart rate when facing the oxidative stress. Practical applications: Polyphenols from tea leaves have been viewed as an antioxidant additive in food, mainly due to the ability of scavenging free radicals and reactive oxygen species. The results of this study suggest that the SS pretreatment could be used as an efficient method to extract TPs from the tea waste for the prevention of oxidative damage in the mouse liver mitochondria and zebrafish embryos. [ABSTRACT FROM AUTHOR]

Abstract P-glycoprotein (P-gp or ABCB1) and multidrug resistance associated proteins (MRPs or ABCC) belonging to the family of ATP-binding cassette (ABC) transporters protect aquatic organisms from various toxicants and pathogen exposure. The function of ABC transporters of Sinonovacula constricta in response to pathogens was explored by cloning the complete cDNA of ABCB1 and ABCC5 of S. constricta through the rapid amplification of cDNA ends. Tissue-specific expression showed that ABCB1 and ABCC5 have the highest expression in hemocytes and gills among eight examined tissues, respectively. The transport activities of ABCB1 and ABCC5 in the gills were severely inhibited after their corresponding inhibitor treatments. The expression levels and transport activities of ABCB1 and ABCB5 were also investigated by using S. constricta samples infected by Vibrio parahaemolyticus at a final concentration of 107 CFU/mL. Results showed that the mRNA expression levels of ABCB1 and ABCC5 were significantly down-regulated when exposed to V. parahaemolyticus at 6 h and 12 h (P < 0.05) and then recovered to normal level at 24 h and 48 h. Consistently, the activities of the two ABC transporters display the same trends as the mRNA expression levels. The results, combined with that of previous investigations, indicated that these activities may be an innate immune response in S. constricta. Immunohistochemical results justified the tissue localization of P-gp in the interface between external environment and tissues. Taken together, our findings demonstrated that ABC transporters form an active, physiological barrier at the tissue–environment interface in S. constricta gills in response to pathogen infection, and they might play a vital role in adaptation and response to environmental stress and the immune defense. Highlights • We firstly identified the full-length transcripts of ABCB1 and long open reading frame of ABCC5 of Sinonovacula constricta. • Hemocytes and gills were the dominant expressed tissues for ABCB1 and ABCC5 respectively. • The mRNA level and activities of MXR proteins in gills were suppressed and then recovered under pathogen infection. • The immunohistochemical results showed that ABCB1 in S. constricta gills was concentrated in tissue-environment interface. [ABSTRACT FROM AUTHOR]