Genomics and tumor microenvironment of breast mucoepidermoid carcinoma based on whole-exome and RNA sequencing.

Mammary mucoepidermoid carcinoma (MEC) is a rare entity. The molecular characteristics of breast MEC have not been fully investigated due to its rarity. We performed a retrospective study among 1000 patients with breast carcinomas and identified four cases of breast MEC. Clinical and demographic data were collected. Immunohistochemistry panels which were used to diagnose salivary gland MEC and breast carcinomas were also performed. MAML2 rearrangements were detected by FISH and fusion partners were identified by RNA sequencing. Whole-exome sequencing (WES) was used to reveal the genomes of these four breast MEC. Then, the biological functions and features of breast MEC were further compared with those of invasive breast carcinomas and salivary gland MEC.According to Ellis and Auclair's methods, these four breast MEC could be classified as low-grade breast MEC. All the patients were alive, and disease-free survival (PFS) ranged from 20 months to 67 months. Among these four breast MEC, two cases were triple-negative, and the other two cases were found to be ER positive, with one also showing HER2 equivocal by immunohistochemical staining, but no amplification in FISH. FISH analysis confirmed the presence of the MAML2 translocation in three of four tumors, and CRTC1-MAML2 fusion was confirmed in two of them by RNA-sequencing. The average coverage size of WES for the tumor mutation burden estimation was 32 Mb. MUC4, RP1L1 and QRICH2 mutations were identified in at least three tumors, and these mutation also existed in breast invasive carcinoma databases (TCGA, Cell 2015; TCGA, Nature 2012). The results showed that there were many genes in breast MEC overlapping with the breast invasive carcinoma databases mentioned above, range from 5 to 63 genes (median:21 genes). Next, we assessed immune cell infiltration levels in these tumors. In all these tumors, M2 macrophages and plasma cell were in the high infiltration group. Our breast MEC showed different results from the salivary gland MEC, whose plasma cells were in the low infiltration group. Overall, we first analyzed the genomics and tumor microenvironment of breast mucoepidermoid carcinoma and proposed our hypothesis that although MECs arising in the breast resemble their salivary gland counterparts phenotypically, our findings indicate that breast MECs probably resemble invasive breast carcinomas at the genetic level and immune cell infiltration levels. More cases and in deep research need to be done to further understand this rare carcinoma.

Effects of light intensity and salinity on formation and performance of microalgal-bacterial granular sludge.

Photosynthetic microorganisms in microalgal-bacterial granular sludge offer advantages in wastewater treatment processes. This study examined the effects of light intensity and salinity on microalgal-bacterial granular sludge formation and microbial changes. Activated sludge was inoculated into three bioreactors and operated in batch treatment mode for 100 days under different light intensities (0, 60, and 120 µmol m-2 s-1) and staged increases in salinity concentration (0, 1, 2, and 3%). Results showed that microalgal-bacterial granular sludge was successfully formed within 30 days, and high light exposure increased algal particle stability and inorganic nitrogen removal (63, 66, 71%), while chemical oxygen demand removal (>95%) was similar across groups. High-throughput sequencing results showed that the critical algae were Chlorella and diatoms, while the main bacteria included Paracoccus and Xanthomarina with high extracellular polymeric substance production. This study aims to enhance the comprehension of MBGS processes in saline wastewater treatment under varying light intensities.

Treatment and outcome of a patient with low-energy 'Logsplitter' injury.

The low-energy 'Logsplitter' fracture, caused by a sprain or fall, is characterized by an intact or slightly separated inferior tibiofibular joint. Compared with the high-energy 'Logsplitter' fracture, this atypical subtype is rarely seen and is easily missed. Here, the case of a 33-year-old male patient with a fractured right ankle as a result of a sprain during walking is reported. The patient initially received routine surgical treatment comprising internal fixation of the fibular, medial and posterior malleoli. Unexpectedly, post-surgery imaging examinations revealed that the medial clear space of the right ankle had widened to 6 mm, due to incomplete reduction of the lateral malleolus, shortening and rotation of the fibula, and an unreduced avulsion fracture block of the anterior malleolus. A revision surgery was then performed to anatomically reduce and fix the lateral malleolus, as well as the anterior malleolus avulsion fracture. During 5 months following surgery, the patient achieved good fracture union and functional restoration of the right ankle. For this rare injury, the present case demonstrates that complete restoration of the fracture is required to achieve good clinical efficacy.

Cervical adenofibroma without clinical symptoms: report of a rare case.

Adenofibroma is an extremely rare benign biphasic tumour composed of glandular and fibrous tissues. It occurs more often in the endometrium but it can also occur in the cervix and extrauterine sites. This case report describes a 39-year-old asymptomatic woman with cervical adenofibroma. The patient was treated successfully with surgical removal of the tumour. As adenofibromas are very rare, the report is presented with a brief review of the literature.

Characterization of a new selenoprotein methionine sulfoxide reductase from Haematococcus pluvialis and its antioxidant activity in response to high light intensity, hydrogen peroxide, glyphosate, and cadmium exposure.

Selenium incorporates into selenocysteine (Sec) which is a key component of selenoproteins implicated in antioxidant defense and redox homeostasis. Methionine sulfoxide reductases (Msr) play crucial roles in cellular defense against environmental stress. Whereas mammals have the MsrB selenoprotein form, unicellular organisms have MsrA. The Sec residue at the conserved catalytic sites of selenoprotein MsrA confers a metabolic advantage over the non-selenoprotein type MsrA. In the present study, the novel selenoprotein HpMsrA from Haematococcus pluvialis was cloned by the rapid amplification of cDNA ends and transformed into the model green alga Chlamydomonas reinhardtii. Alignment of homologs revealed the presence of the conserved catalytic domain GUFW and showed that the HpMsrA protein comprises Sec (U) at the N-terminus but no recycled Cys at the C-terminus. We studied the response of HpMsrA expression to selenite, high light intensity, hydrogen peroxide, cadmium nitrate, and glyphosate exposure via real-time quantitative PCR and enzyme activity analysis. The results demonstrated that HpMsrA protects cellular proteins against oxidative and environmental stressors. Compared with wild type C. reinhardtii, the transformant exhibited a superior antioxidant ability. The discoveries made herein shed light on the antioxidant physiology and environmental stress resistance mechanisms of the selenoproteins in microalgae. This information may aid in conducting environmental risk assessments of aquatic ecosystems involving microalgae known to respond rapidly and quantitatively to abiotic stress factors promoting excessive reactive oxygen species generation.

Toxicological Effects of Microplastics and Sulfadiazine on the Microalgae Chlamydomonas reinhardtii.

Despite the fact that microplastics (MPs) facilitate the adsorption of environmental organic pollutants and influence their toxicity for organisms, more study is needed on the combination of MPs and antibiotics pollutant effects. In this study, polystyrene MPs (1 and 5 µm) and sulfadiazine (SDZ) were examined separately and in combination on freshwater microalga, Chlamydomonas reinhardtii. The results suggest that both the MPs and SDZ alone and in combination inhibited the growth of microalgae with an increasing concentration of MPs and SDZ (5-200 mg l-1); however, the inhibition rate was reduced by combination. Upon exposure for 7 days, both the MPs and SDZ inhibited algal growth, reduced chlorophyll content, and enhanced superoxide dismutase (SOD) activities, whereas glutathione peroxidase (GSH-Px) activity was elevated only with the exposure of 1 µm MPs. Fluorescence microscopy and scanning electron microscopy also indicated that particle size contributed to the combined toxicity by aggregating MPs with periphery pollutants. Further, the amount of extracellular secretory protein increased in the presence of MPs and SDZ removal ratio decreased when MPs and SDZ coexisted, suggesting that MPs affected SDZ metabolism by microalgae. The particle size of microplastics affected the toxicity of MPs on microalgae and the combined effect of MPs and SDZ could be mitigated by MPs adsorption. These findings provide insight into microalgae responses to the combination of MPs and antibiotics in water ecosystems.

Efficacy and safety of a live attenuated influenza vaccine in Chinese healthy children aged 3-17 years in one study center of a randomized, double-blind, placebo-controlled phase 3 clinical trial, 2016/17 season.

BACKGROUND: No data on the safety and efficacy of a live attenuated influenza vaccine in China have ever been reported. METHODS: At a site of a phase 3 randomized, double-blind, placebo-controlled clinical trial in eastern China, eligible healthy children aged 3-17 years underwent randomization to receive live attenuated vaccine or placebo at a ratio of 1:1. The primary objective of the study was the prevention of laboratory-confirmed influenza illness during the surveillance period, starting on day 15 after vaccination. RESULTS: A total of 2000 participants were enrolled, with 998 receiving the vaccine and 1001 receiving placebo. Sixty-four cases of influenza-like illness were observed, of which, 44 were laboratory-confirmed (12 in vaccine group versus 32 in placebo group). Vaccine efficacy was 62.5% (95%CI: 27.6-80.6) against all types of influenza and 63.3% (95%CI: 27.5-81.5) against influenza H3N2 illness. 11 severe adverse events reported (7 in LAIV group versus 4 in placebo group) were all deemed to be non-vaccine-related. Adverse events occurred in 412 (41.3%) participants in the vaccine group versus 389 (38.9%; p = 0.274) participants in the placebo group. Significant increase incidence of fever was observed in participants in the vaccine group, especially in those aged 3-9 years. CONCLUSIONS: The live attenuated influenza vaccine showed good efficacy and safety among 3- to 17-year-olds children during the 2016-2017 season at a site in eastern China. Clinical Trial Registry Number: NCT02964065.

The EIL transcription factor family in soybean: Genome-wide identification, expression profiling and genetic diversity analysis.

The ETHYLENE INSENSITIVE3-LIKE (EIL) transcription factor family plays a critical role in the ethylene signaling pathway, which regulates a broad spectrum of plant growth and developmental processes, as well as defenses to myriad stresses. Although genome-wide analysis of this family has been carried out for several plant species, no comprehensive analysis of the EIL gene family in soybean has been reported so far. Furthermore, there are few studies on the functions of EIL genes in soybean. In this study, we identified 12 soybean (Gm) EIL genes, which we divided into three groups based on their phylogenetic relationships. We then detected their duplication status and found that most of the GmEIL genes have duplicated copies derived from two whole-genome duplication events. These duplicated genes underwent strong negative selection during evolution. We further analyzed the transcript profiles of GmEIL genes using the transcriptome data and found that their spatio-temporal and stress expression patterns varied considerably. For example, GmEIL1-GmEIL5 were found to be strongly expressed in almost every sample, while GmEIL8-GmEIL12 exhibited low expression, or were not expressed at all. Additionally, these genes showed different responses to dehydration, salinity and phosphate starvation. Finally, we surveyed genetic variations of these genes in 302 resequenced wild soybeans, landraces and improved soybean cultivars. Our data showed that most GmEIL genes are well conserved, and are not modified in domesticated or improved cultivars. Together, these findings provide a potentially valuable resource for characterizing the GmEIL gene family and lay the basis for further elucidation of their molecular mechanisms.

Genome and Transcriptome Sequencing of the Astaxanthin-Producing Green Microalga, Haematococcus pluvialis.

Haematococcus pluvialis is a freshwater species of Chlorophyta, family Haematococcaceae. It is well known for its capacity to synthesize high amounts of astaxanthin, which is a strong antioxidant that has been utilized in aquaculture and cosmetics. To improve astaxanthin yield and to establish genetic resources for H. pluvialis, we performed whole-genome sequencing, assembly, and annotation of this green microalga. A total of 83.1 Gb of raw reads were sequenced. After filtering the raw reads, we subsequently generated a draft assembly with a genome size of 669.0 Mb, a scaffold N50 of 288.6 kb, and predicted 18,545 genes. We also established a robust phylogenetic tree from 14 representative algae species. With additional transcriptome data, we revealed some novel potential genes that are involved in the synthesis, accumulation, and regulation of astaxanthin production. In addition, we generated an isoform-level reference transcriptome set of 18,483 transcripts with high confidence. Alternative splicing analysis demonstrated that intron retention is the most frequent mode. In summary, we report the first draft genome of H. pluvialis. These genomic resources along with transcriptomic data provide a solid foundation for the discovery of the genetic basis for theoretical and commercial astaxanthin enrichment.

Two new species of freshwater Macrostomum (Rhabditophora: Macrostomorpha) found in China.

There is limited information about the ecology of freshwater flatworms in China, in particular, the members of the genus Macrostomum. Surveying freshwater bodies to explore the species diversity is the first step to obtain more ecological information about those flatworms. We hereby report the discovery of two new species of freshwater flatworms in Macrostomum. Macrostomum chongqingensis Lin and Wang, n. sp. was discovered from stones in the shallow water of Jialing River bed near Chongqing. Macrostomum zhaoqingensis Lin and Wang, n. sp. was discovered in the water of Xing Lake near Zhaoqing city, Guangdong province. Xing Lake is a part of an ancient river that has changed its course. For each species, we describe the morphology of the whole specimen, penis stylet, and sperm, as well as other internal anatomical features based on serial sections. We determined 18S, 28S rDNA and COI sequences from the two new species, and performed a phylogenetic analysis to confirm the status and evolutionary position of the two new species. We discuss the significance of our discoveries in terms of flatworm biodiversity studies in China and the Oriental realm.

Effects of selenite on green microalga Haematococcus pluvialis: Bioaccumulation of selenium and enhancement of astaxanthin production.

Algae are at a low trophic level and play a crucial role in aquatic food webs. They can uptake and accumulate the trace element selenium (Se), which can be either essential or toxic to algal growth depending on the dosage and species. Se toxicity and algae resistance varied across different organisms. In order to investigate the effects of Se on the unicellular green alga Haematococcus pluvialis, an important industrial resource for natural astaxanthin, the algal growth rate, chlorophyll content, and fluorescence parameters were derived from experimental treatment with different concentrations of selenite. The results showed that the EC50 for the algal growth rate was 24mg/L, and that a low dosage of selenite (3mg/L) may not hinder H. pluvialis cell growth, but selenite at levels higher than 13mg/L do restrain cell growth. Bioaccumulation experiments showed that H. pluvialis accumulated up to 646µg/g total Se and 380µg/g organic Se, dry weight. However, treatment with high concentrations of selenite significantly increased intracellular hydrogen peroxide levels, antioxidant enzyme activity, and the production of astaxanthin, suggesting that Se bioaccumulation might be toxic to H. pluvialis.

Gorgeous mosaic of mitochondrial genes created by horizontal transfer and gene conversion.

The best known outcome of horizontal gene transfer (HGT) is the introduction of novel genes, but other outcomes have been described. When a transferred gene has a homolog in the recipient genome, the native gene may be functionally replaced (and subsequently lost) or partially overwritten by gene conversion with transiently present foreign DNA. Here we report the discovery, in two lineages of plant mitochondrial genes, of novel gene combinations that arose by conversion between coresident native and foreign homologs. These lineages have undergone intricate conversion between native and foreign copies, with conversion occurring repeatedly and differentially over the course of speciation, leading to radiations of mosaic genes involved in respiration and intron splicing. Based on these findings, we develop a model--the duplicative HGT and differential gene conversion model--that integrates HGT and ongoing gene conversion in the context of speciation. Finally, we show that one of these HGT-driven gene-conversional radiations followed two additional types of conversional chimerism, namely, intramitochondrial retroprocessing and interorganellar gene conversion across the 2 billion year divide between mitochondria and chloroplasts. These findings expand our appreciation of HGT and gene conversion as creative evolutionary forces, establish plant mitochondria as a premiere system for studying the evolutionary dynamics of HGT and its genetic reverberations, and recommend careful examination of bacterial and other genomes for similar, likely overlooked phenomena.

Deletion of phospholipase C beta4 in thalamocortical relay nucleus leads to absence seizures.

Absence seizures are characterized by cortical spike-wave discharges (SWDs) on electroencephalography, often accompanied by a shift in the firing pattern of thalamocortical (TC) neurons from tonic to burst firing driven by T-type Ca(2+) currents. We recently demonstrated that the phospholipase C beta4 (PLCbeta4) pathway tunes the firing mode of TC neurons via the simultaneous regulation of T- and L-type Ca(2+) currents, which prompted us to investigate the contribution of TC firing modes to absence seizures. PLCbeta4-deficient TC neurons were readily shifted to the oscillatory burst firing mode after a slight hyperpolarization of membrane potential. TC-limited knockdown as well as whole-animal knockout of PLCbeta4 induced spontaneous SWDs with simultaneous behavioral arrests and increased the susceptibility to drug-induced SWDs, indicating that the deletion of thalamic PLCbeta4 leads to the genesis of absence seizures. The SWDs were effectively suppressed by thalamic infusion of a T-type, but not an L-type, Ca(2+) channel blocker. These results reveal a primary role of TC neurons in the genesis of absence seizures and provide strong evidence that an alteration of the firing property of TC neurons is sufficient to generate absence seizures. Our study presents PLCbeta4-deficient mice as a potential animal model for absence seizures.