Photobody formation spatially segregates two opposing phytochrome B signaling actions of PIF5 degradation and stabilization.

Photoactivation of the plant photoreceptor and thermosensor phytochrome B (PHYB) triggers its condensation into subnuclear membraneless organelles named photobodies (PBs). However, the function of PBs in PHYB signaling remains frustratingly elusive. Here, we found that PHYB recruits PHYTOCHROME-INTERACTING FACTOR 5 (PIF5) to PBs. Surprisingly, PHYB exerts opposing roles in degrading and stabilizing PIF5. Perturbing PB size by overproducing PHYB provoked a biphasic PIF5 response: while a moderate increase in PHYB enhanced PIF5 degradation, further elevating the PHYB level stabilized PIF5 by retaining more of it in enlarged PBs. Conversely, reducing PB size by dim light, which enhanced PB dynamics and nucleoplasmic PHYB and PIF5, switched the balance towards PIF5 degradation. Together, these results reveal that PB formation spatially segregates two antagonistic PHYB signaling actions - PIF5 stabilization in PBs and PIF5 degradation in the surrounding nucleoplasm - which could enable an environmentally sensitive, counterbalancing mechanism to titrate nucleoplasmic PIF5 and environmental responses.

Oral health self-management barriers among rural older adults in Guangxi, China: A qualitative study.

BACKGROUND: Objective: To understand the barriers associated with self-management of oral health among rural older adults in Guangxi, and to explore the high incidence of oral problems. This information will assist in the formulation of relevant strategies to solve the oral health problems in this population. METHODS: Taking a phenomenological approach, the current status of, and barriers to, oral health self-management in rural older adults from different regions of Guangxi were explored. Participants were purposively selected and interviewed face-to-face. RESULTS: The interviews yielded four overarching themes and six corresponding sub-themes pertaining to barriers in oral health self-management. These included: (1) Older adults' understanding of oral health and disease, perceptions of oral health and their oral health behaviours; (2) Problems in accessing oral health information; (3) Role of family support; and (4) Barriers to healthcare that included access to dental services, oral treatment experience and financial burden of access to dental care. CONCLUSION: Rural older adults in Guangxi face oral health self-management barriers. Improving access to oral healthcare services and changing existing oral health perceptions and habits may assist them in overcoming self-management challenges.

Dissection of Daytime and Nighttime Thermoresponsive Hypocotyl Elongation in Arabidopsis.

Hypocotyl elongation in Arabidopsis is widely utilized as a readout for phytochrome B (phyB) signaling and thermomorphogenesis. Hypocotyl elongation is gated by the circadian clock and, therefore, it occurs at distinct times depending on day length or seasonal cues. In short-day conditions, hypocotyl elongation occurs mainly at the end of nighttime when phyB reverts to the inactive form. In contrast, in long-day conditions, hypocotyl elongation occurs during the daytime when phyB is in the photoactivated form. Warm temperatures can induce hypocotyl growth in both long-day and short-day conditions. However, the corresponding daytime and nighttime temperature responses reflect distinct underpinning mechanisms. Here, we describe assays for dissecting the mechanisms between daytime and nighttime thermoresponsive hypocotyl elongation.

Photobody formation spatially segregates two opposing phytochrome B signaling actions to titrate plant environmental responses.

Photoactivation of the plant photoreceptor and thermosensor phytochrome B (PHYB) triggers its condensation into subnuclear photobodies (PBs). However, the function of PBs remains frustratingly elusive. Here, we found that PHYB recruits PHYTOCHROME-INTERACTING FACTOR5 (PIF5) to PBs. Surprisingly, PHYB exerts opposing roles in degrading and stabilizing PIF5. Perturbing PB size by overproducing PHYB provoked a biphasic PIF5 response: while a moderate increase in PHYB enhanced PIF5 degradation, further elevating the PHYB level stabilized PIF5 by retaining more of it in enlarged PBs. These results reveal a PB-mediated light and temperature sensing mechanism, in which PHYB condensation confers the co-occurrence and competition of two antagonistic phase-separated PHYB signaling actions-PIF5 stabilization in PBs and PIF5 degradation in the surrounding nucleoplasm-thereby enabling an environmentally-sensitive counterbalancing mechanism to titrate nucleoplasmic PIF5 and its transcriptional output. This PB-enabled signaling mechanism provides a framework for regulating a plethora of PHYB-interacting signaling molecules in diverse plant environmental responses.

Co-Culture of White Rot Fungi Pleurotus ostreatus P5 and Bacillus amyloliquefaciens B2: A Strategy to Enhance Lipopeptide Production and Suppress of Fusarium Wilt of Cucumber.

Fusarium wilt, caused by Fusarium oxysporum f. sp. cucumerinum (FOC), poses a serious threat to cucumber productivity. Compared to traditional chemical pesticides, biological control strategies have attracted more attention recently owing to their effectiveness against pathogens and their environmental safety. This study investigated the effect of white rot fungi Pleurotus ostreatus P5 on the production of cyclic lipopeptides (CLPs) of Bacillus amyloliquefaciens B2 and the potential co-culture filtrate of strains B2 and P5 to control cucumber Fusarium wilt. A PCR amplification of CLP genes revealed that B. amyloliquefaciens B2 had two antibiotic biosynthesis genes, namely, ituA and srf, which are involved in iturin A and surfactin synthesis. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) revealed that CLPs derived from strain B2 contained two families, iturin A (C14, C15) and surfactin (C12-C17). The co-culture exhibited an enhanced accumulation of iturin A and surfactin compared to the monoculture of strain B2. Furthermore, the gene expressions of ituA and srf were both significantly upregulated when co-cultured with the fungus compared to monocultures. In an in vitro experiment, the co-culture filtrate and monoculture filtrate of B. amyloliquefaciens B2 inhibited mycelial growth by 48.2% and 33.2%, respectively. In a greenhouse experiment, the co-culture filtrate was superior to the monoculture filtrate in controlling cucumber Fusarium wilt disease and in the promotion of plant growth. Co-culture filtrate treatment significantly enhanced the microbial metabolic activity and decreased the abundance of FOC in the rhizosphere soil. These results show that the co-culture of P. ostreatus P5 and B. amyloliquefaciens B2 has great potential in cucumber Fusarium wilt disease prevention by enhancing the production of bacterial CLPs.

The noncoding RNA HIDDEN TREASURE 1 promotes phytochrome B-dependent seed germination by repressing abscisic acid biosynthesis.

Light is a major environmental factor for seed germination. Red light-activated phytochrome B (phyB) promotes seed germination by modulating the dynamic balance of two phytohormones, gibberellic acid (GA) and abscisic acid (ABA). How phyB modulates ABA biosynthesis after perceiving a light signal is not yet well understood. Here, we identified the noncoding RNA HIDDEN TREASURE 1 (HID1) as a repressor of ABA biosynthesis acting downstream of phyB during Arabidopsis thaliana seed germination. Loss of HID1 function led to delayed phyB-dependent seed germination. Photoactivated phyB promoted the accumulation of HID1 in the radicle within 48 h of imbibition. Our transcriptomics analysis showed that HID1 and phyB co-regulate the transcription of a common set of genes involved in ABA and GA metabolism. Through a forward genetic screen, we identified three ABA biosynthesis genes, ABA DEFICIENT 1 (ABA1), ABA2, and ABA3, as suppressors of HID1. We further demonstrated that HID1 directly inhibits the transcription of 9-CIS-EPOXYCAROTENOID DIOXYGENASE (NCED9), a gene encoding a key rate-limiting enzyme of ABA biosynthesis. HID1 interacts with ARABIDOPSIS TRITHORAX-RELATED7 (ATXR7), an H3K4me3 methyltransferase, inhibiting its occupancy and H3K4me3 modification at the NCED9 locus. Our study reveals a nuclear mechanism of phyB signaling transmitted through HID1 to control the internal homeostasis of ABA and GA, which gradually optimizes the transcriptional network during seed germination.

Aerobic exercise regulates FGF21 and NLRP3 inflammasome-mediated pyroptosis and inhibits atherosclerosis in mice.

Fibroblast growth factor 21 (FGF21), a known risk factor for atherosclerosis, is readily regulated by exercise, and it can inhibit NOD-like receptor protein 3 (NLRP3)-mediated pyroptosis. However, it is not clear whether aerobic exercise inhibits atherosclerosis via these pathways. Eight-week-old apolipoprotein E-deficient (ApoE-/-) mice on a high-fat diet were randomly divided into 1-h post-exercise (EX-1h), 24-h post-exercise (EX-24h), and sedentary (SED) groups. C57BL/6J wild-type mice fed normal chow served as controls (WT group). Mice in the EX-1h and EX-24h groups were subjected to treadmill exercise training for 12 weeks. Aerobic exercise reduced body weight; blood glucose, lipid, and inflammation levels; and aortic plaque area proportion. Aerobic exercise increased the sensitivity of FGF21 by upregulating the expression of the downstream receptor adiponectin (ApN); the serum FGF21 level after exercise increased initially, and then decreased. Aerobic exercise downregulated the expression of NLRP3 inflammasome-mediated pyroptosis-related markers in the aorta, and FGF21 may participate in the above process. Meanwhile, the liver may be the tissue source of serum FGF21 during aerobic exercise. In conclusion, aerobic exercise may inhibit atherogenesis by regulating FGF21 and NLRP3 inflammasome-mediated pyroptosis. Our study provides new information on the atherosclerosis-preventing mechanism of aerobic exercise.

The Arabidopsis DREAM complex antagonizes WDR5A to modulate histone H3K4me2/3 deposition for a subset of genome repression.

The master transcriptional repressor DREAM (dimerization partner, RB-like, E2F and multivulval class B) complex regulates the cell cycle in eukaryotes, but much remains unknown about how it transmits repressive signals on chromatin to the primary transcriptional machinery (e.g., RNA polymerase II [Pol II]). Through a forward genetic screen, we identified BTE1 (barrier of transcription elongation 1), a plant-specific component of the DREAM complex. The subsequent characterization demonstrated that DREAM complex containing BTE1 antagonizes the activity of Complex Proteins Associated with Set1 (COMPASS)-like complex to repress H3K4me3 occupancy and inhibits Pol II elongation at DREAM target genes. We showed that BTE1 is recruited to chromatin at the promoter-proximal regions of target genes by E2F transcription factors. DREAM target genes exhibit characteristic enrichment of H2A.Z and H3K4me2 modification on chromatin. We further showed that BTE1 directly interacts with WDR5A, a core component of COMPASS-like complex, repressing WDR5A chromatin binding and the elongation of transcription on DREAM target genes. H3K4me3 is known to correlate with the Pol II transcription activation and promotes efficient elongation. Thus, our study illustrates a transcriptional repression mechanism by which the DREAM complex dampens H3K4me3 deposition at a set of genes through its interaction with WDR5A.

Development and evaluation of a meat mitochondrial metagenomic (3MG) method for composition determination of meat from fifteen mammalian and avian species.

BACKGROUND: Bioassessment and biomonitoring of meat products are aimed at identifying and quantifying adulterants and contaminants, such as meat from unexpected sources and microbes. Several methods for determining the biological composition of mixed samples have been used, including metabarcoding, metagenomics and mitochondrial metagenomics. In this study, we aimed to develop a method based on next-generation DNA sequencing to estimate samples that might contain meat from 15 mammalian and avian species that are commonly related to meat bioassessment and biomonitoring. RESULTS: In this project, we found the meat composition from 15 species could not be identified with the metabarcoding approach because of the lack of universal primers or insufficient discrimination power. Consequently, we developed and evaluated a meat mitochondrial metagenomics (3MG) method. The 3MG method has four steps: (1) extraction of sequencing reads from mitochondrial genomes (mitogenomes); (2) assembly of mitogenomes; (3) mapping of mitochondrial reads to the assembled mitogenomes; and (4) biomass estimation based on the number of uniquely mapped reads. The method was implemented in a python script called 3MG. The analysis of simulated datasets showed that the method can determine contaminant composition at a proportion of 2% and the relative error was < 5%. To evaluate the performance of 3MG, we constructed and analysed mixed samples derived from 15 animal species in equal mass. Then, we constructed and analysed mixed samples derived from two animal species (pork and chicken) in different ratios. DNAs were extracted and used in constructing 21 libraries for next-generation sequencing. The analysis of the 15 species mix with the method showed the successful identification of 12 of the 15 (80%) animal species tested. The analysis of the mixed samples of the two species revealed correlation coefficients of 0.98 for pork and 0.98 for chicken between the number of uniquely mapped reads and the mass proportion. CONCLUSION: To the best of our knowledge, this study is the first to demonstrate the potential of the non-targeted 3MG method as a tool for accurately estimating biomass in meat mix samples. The method has potential broad applications in meat product safety.

Identification of an Autophagy-Related Gene Signature for the Prediction of Prognosis in Early-Stage Colorectal Cancer.

Purpose: A certain number of early-stage colorectal cancer (CRC) patients suffer tumor recurrence after initial curative resection. In this context, an effective prognostic biomarker model is constantly in need. Autophagy exhibits a dual role in tumorigenesis. Our study aims to develop an autophagy-related gene (ATG) signature-based on high-throughput data analysis for disease-free survival (DFS) prognosis of patients with stage I/II CRC. Methods: Gene expression profiles and clinical information of CRC patients extracted from four public datasets were distributed to discovery and training cohort (GSE39582), validation cohort (TCGA CRC, n = 624), and meta-validation cohort (GSE37892 and GSE14333, n = 420). Autophagy genes significantly associated with prognosis were identified. Results: Among 655 autophagy-related genes, a 10-gene ATG signature, which was significantly associated with DFS in the training cohort (HR, 2.76[1.56-4.82]; p = 2.06 × 10-4), was constructed. The ATG signature, stratifying patients into high and low autophagy risk groups, was validated in the validation (HR, 2.29[1.15-4.55]; p = 1.5 × 10-2) and meta-validation cohorts (HR, 2.5[1.03-6.06]; p = 3.63 × 10-2) and proved to be prognostic in a multivariate analysis. Functional analysis revealed enrichment of several immune/inflammatory pathways in the high autophagy risk group, where increased infiltration of T regulatory cells (Tregs) and decreased infiltration of M1 macrophages were observed. Conclusion: Our study established a prognostic ATG signature that effectively predicted DFS for early-stage CRC patients. Meanwhile, the study also revealed the possible relationship among autophagy process, immune/inflammatory response, and tumorigenesis.

[Abdominal acupoint thread embedding therapy based on "brain-intestinal connection" for mild-to-moderate Alzheimer's disease and its effects on serum levels of APP and Aß1-42].

OBJECTIVE: To compare the clinical efficacy of abdominal acupoint thread embedding therapy based on "brain-intestinal connection" combined with donepezil hydrochloride tablets and oral donepezil hydrochloride tablets alone for mild-to-moderate Alzheimer's disease (AD) and observe its effects on amyloid precursor protein (APP) and ß-amyloid protein1-42 (Aß1-42). METHODS: Sixty patients with AD were randomly divided into an observation group (30 cases, 3 cases dropped off) and a control group (30 cases, 3 cases dropped off). The patients in the control group were treated with donepezil hydrochloride tablets (5 mg per day); based on the treatment in the control group, the patients in the observation group were treated with abdominal acupoint thread embedding therapy at Zhongwan (CV 12), Xiawan (CV 10), Huaroumen (ST 24), Wailing (ST 26), Daheng (SP 15), etc., once every 10 days. Both groups were treated for 2 months. The mini-mental state examination (MMSE), Alzheimer's disease assessment scale-cognitive subscale (ADAS-Cog), activity of daily living scale (ADL), neuropsychiatric inventory questionnaire (NPI) as well as the serum levels of APP and Aß1-42 were observed before and after treatment in the two groups. RESULTS: After treatment, the MMSE scores in the two groups were higher than those before treatment (P<0.05), and the ADAS-Cog, ADL and NPI scores were lower than those before treatment (P<0.05). After treatment, the MMSE score in the observation group was higher than that in the control group (P<0.05), and the ADAS-Cog, ADL and NPI scores were lower than those in the control group (P<0.05). After treatment, the serum levels of APP and Aß1-42 were lower than those before treatment (P<0.05), and the serum levels of APP and Aß1-42 in the observation group was lower than those in the control group (P<0.05). CONCLUSION: The abdominal acupoint thread embedding therapy based on the theory of "brain-intestinal connection" combined with donepezil hydrochloride tablets can improve cognitive function, self-care ability of daily life and mental behavior, and reduce the serum levels of APP and Aß1-42 in patients with mild-to-moderate AD, which have superior clinical effect to donepezil hydrochloride tablets alone.

[Pollution Level, Distribution Characteristic, and Ecological Risk Assessment of Environmentally Persistent Pharmaceutical Pollutants in Surface Water of Jiangsu Province].

Because Jiangsu is an important economic province of China, it is necessary to examine the pollution characteristics and assess the ecological risk of environmentally persistent pharmaceutical pollutants (EPPPs) in this region. In this study, surface water samples were obtained from grade 1-4 rivers and lakes (with an area of 50 km2 or more) in Jiangsu Province, and then analyzed to determine the pollution level of EPPPs. In total, 35 EPPPs were detected in the surface water of Jiangsu Province, with total concentrations in the samples ranging from 66.74 to 2189.83 ng·L-1. The 17 EPPPs with a detection rate of more than 25% are discussed in this study. The total concentrations of 35 EPPPs were 72.48-1142.79 ng·L-1, and the mean concentration was 345.20 ng·L-1. The total concentration of EPPPs was higher in the north and south than in the central part of Jiangsu. Yangzhou city had the highest concentration of EPPPs in the whole province, and the main sources of this pollution were domestic sewage, shipping sewage discharge, and drug use in fishery breeding. The total concentration of EPPPs decreased on both sides of the region, with the Beijing-Hangzhou Canal and waste from the Yellow River forming the middle line. An ecological risk assessment of 17 EPPPs showed that single target drugs posed a low risk to water ecology in Jiangsu Province. The combined risk quotient of 17 EPPPs in water of Jiangsu Province was 0.03-0.52, indicating that EPPPs posed a low to moderate risk.

In vivo biocompatibility evaluation of Zn-0.05Mg-(0, 0.5, 1wt%)Ag implants in New Zealand rabbits.

Bio-absorbable Zn alloys have been attractive replacements for the traditionally permanent implants due to their reasonable mechanical strength and elongation, degradation rate, and biocompatibility. The hybridization addition of Mg and Ag elements could greatly improve the mechanical properties and antibacterial ability of Zn, respectively. In the present paper, in vivo biocompatibility for the Zn-0.05Mg-(0, 0.5, 1 wt%) Ag implants in New Zealand rabbit was qualitatively evaluated during the implantation periods of 4, 12, and 24 weeks. The blood serum biochemical parameters and in vivo integrity of the implants in the live rabbits were monitored by using clinical chemistry analyzing and X-ray radiographic imaging techniques during the implantation process, respectively. There is no great difference in the serum biochemical indicator between the implanted rabbits and the control group. Especially the levels of serum Zn and serum Mg normalize after implantation of 24 weeks. The interfacial adherence between the implants and newly formed bones, and the histopathological morphology of heart, liver, and kidney were observed morphologically under the microscope. The new bones formed and grew surrounding the implants after 12 weeks' post-operation, which were well joined with the original cortical bones after post-implantation of 24 weeks. The heart, liver and kidney were not negatively influenced as evidenced from the serum biochemical indicators and morphologies of the tissues. Zn-0.05Mg-(0, 0.5, 1 wt%) Ag alloys are proved to be in vivo biocompatible and potential candidates for the biodegradable medical implants.

Berberine ameliorates obesity-induced chronic inflammation through suppression of ER stress and promotion of macrophage M2 polarization at least partly via downregulating lncRNA Gomafu.

BACKGROUND: Berberine has been established as a potential drug for inflammation and metabolic disorder. Here, we aimed to explore the effects and the underlying mechanisms of berberine on obesity-induced chronic inflammation. METHODS: Mice were fed with high-fat diet to induce obesity. Inflammation in adipocytes were induced with treatment of free fatty acids. The expression of IL-4, CD206, ARG1 and other markers were used to identify M1 and M2 polarization. The expression of GPR78 and CHOP were used to evaluate endoplasmic reticulum stress. H&E staining was used to reveal the adipose tissue macrophage and adipocytes enlargement. RESULTS: Berberine treatment attenuated endoplasmic reticulum stress and inflammation in obese mice and free fatty acids-treated adipocytes. Overexpression of lncRNA Gomafu partially blocked the protective effects of berberine in free fatty acids-treated adipocytes by increasing endoplasmic reticulum stress. Moreover, Gomafu overexpression partly reversed berberine-induced enhancement of M2 polarization in macrophages. Finally, Gomafu overexpression induced ER stress and inflammation in mice, which were improved by berberine administration. CONCLUSIONS: Berberine improves obesity-induced chronic inflammation by alleviating endoplasmic reticulum stress and consequently promoting macrophage M2 polarization. And these protective effects were mediated at least partly by the suppression of lncRNA Gomafu.

Integrated fate assessment of aromatic amines in aerobic sewage treatment plants.

The fate and exposure of chemicals in sewage treatment plants (STPs) are major considerations in risk assessment and environmental regulation. The biodegradability and removal of seven aromatic amines were systematically evaluated using a three-tiered integrated method: a standard ready biodegradability test, an aerobic sewage treatment simulation method, and model prediction. In tier 1, the seven aromatic amines were not readily biodegraded after 28 days. In adapted aerobic active sludge, 4-isopropyl aniline, 2,4-diaminotoluene, and 4-nitroaniline among them exhibited the degradation half-life time less than 20 h, the other four aromatic amines exhibited persistent with degradation half-life of > 60 h. In tier 2 of the aerobic sewage treatment simulation testing, 2,4-diaminotoluene, 4-nitroaniline, and 4-isopropylaniline demonstrated moderately to high overall removal. Hydraulic retention time (HRT) affects the removal with the optimum HRT was determined to be 12 h to 24. 2,6-Dimethyl aniline, 2-chloro-4-nitroaniline, 2,6-diethylaniline, and 3,4-dichloroaniline were not removed during the test, indicting these four aromatic amines will enter surface water and hence pose a potential risk to aquatic ecology. Considering the lack of an STP model in China for regulation purposes, in tier 3, we developed a Chinese STP (aerobic) (abbreviated as C-STP(O)) model that reflects a universal scenario for China to predict the fate. The predicted degradation, volatilization, and absorption showed a close relationship to the physicochemical properties of the chemicals, and had same tendency with tier 2 simulation test. The prediction showed that biodegradation rather than absorption or volatilization was the main removal process of aromatic amines in aerobic STP. With the combination of modified kinetics test with C-STP (O) model, the chemical fate can be more accurately predicted than using only the readily biodegradation result.

Lysobacter alkalisoli sp. nov., a chitin-degrading strain isolated from saline-alkaline soil.

Strains of Lysobacter, thought to play vital roles in the environment for their high enzyme production capacity, are ubiquitous in various ecosystems. During an analysis of bacterial diversity in saline soil, a Gram-stain-negative, aerobic, chitin-degrading bacterial strain, designated SJ-36T, was isolated from saline-alkaline soil sampled at Tumd Right Banner, Inner Mongolia, PR China. Strain SJ-36T grew at 4-40 °C (optimum, 30 °C), pH 5.0-10.0 (optimum, pH 7.0-8.0) and 0-6 % NaCl (optimum, 1.0 %). Oxidase and catalase activities were positive. A phylogenetic tree based on 16S rRNA gene sequences and the phylogenomic tree both showed that strain SJ-36T formed a tight clade with Lysobacter maris KMU-14T (sharing 97.6 % 16S rRNA gene similarity) and Lysobacter aestuarii S2-CT (97.8 %). The major polar lipids of strain SJ-36T were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, two unidentified lipids and one unidentified phospholipid. The major fatty acids were iso-C15 : 0 (37.5 %), summed feature 9 (14.0 %; iso-C17 : 1ω9c and/or C16 : 0 10-methyl) and iso-C11 : 0 (10.6 %). Q-8 was the predominant ubiquinone. Its genomic DNA G+C content was 66.6 mol%. The average nucleotide identity values of strain SJ-36T to L. maris KMU-14T, L. aestuarii S2-CT and other type strains were 81.5, 79.1 and <79.0 %, respectively. The results of physiological, phenotypic and phylogenetic characterizations allowed the discrimination of strain SJ-36T from its phylogenetic relatives. Lysobacter alkalisoli sp. nov. is therefore proposed with strain SJ-36T (=CGMCC 1.16756T=KCTC 43039T) as the type strain.

Development of an integral strategy for non-target and target analysis of site-specific potential contaminants in surface water: A case study of Dianshan Lake, China.

Surface water contains a large number of potential pollutants and their transformation products, which cannot be discovered by normal target analysis alone. To detect site-specific and unknown contaminants in the environment, we established an integral analytical strategy based on liquid chromatography-high resolution mass spectrometry (LC-HRMS) combined with data processing using specific software (Compound Discoverer 3.0). In this case study of Dianshan Lake, 95 potential contaminants were tentatively identified and ranked by the scoring system. Then, the 95 compounds were categorized into 4 subgroups: pesticides, drugs, plastic additives and surfactants. To determine the sources and distribution of those pollutants, 4 heat maps were developed based on the sum of peak areas of respective categories. In addition, 19 substances with high exposure risk among the 95 compounds tentatively identified were confirmed and quantified. In the present study, the analytical strategy with non-target screening followed by target analysis demonstrated that pesticides and plastic additives are the two dominant types of contaminants in Dianshan Lake. High accuracy and high-resolution data combined with integrated software provided abundant information for the identification of a wide range of potential contaminants in the environment. This approach can be a useful tool for the simple and rapid screening and tentative detection of site-specific contaminants.

RARRES1 is a novel immune-related biomarker in GBM.

Immunotherapy is a promising route for the treatment of glioblastoma (GBM). Researchers have conducted a large number of studies on the pathogenesis of GBM; however, these studies are not comprehensive. High-throughput sequence analysis allows for insights into the pathogenesis of GBM. In this study, we used The Cancer Genome Atlas dataset to identify the function of RARRES1 enriched in GBM, especially in the WHO grade-IV cases. We discovered that RARRES1 is highly expressed in patients with mesenchymal subtype, unmethylated MGMT, IDH1 wild type, and non-G-CIMP, all of which are molecular characteristics of malignant GBM. Results of the immune microenvironment analysis showed that RARRES1 is strongly correlated with dendritic cells PD1, PDL2, TIM3, and CTLA4, which are the immune checkpoints in GBM. Furthermore, according to the overall survival and status analysis, a high expression of RARRES1 was found to be an unfavorable factor for prognosis. This indicates that RARRES1 may participate in the pathogenesis and immune-related processes in GBM, and may serve as a therapeutic target.

Participation of tumor suppressors long non-coding RNA MEG3, microRNA-377 and PTEN in glioma cell invasion and migration.

PURPOSE: Glioma is a common and fatal intracranial tumor. Both miR-377 and lncRNA MEG3 are tumor suppressors. This study was performed to investigate the association between miR-377 and lncRNA MEG3 in glioma cells. METHODS: U118 and U251 cell lines were incubated in Dulbecco's modified Eagle's medium supplemented with miR-377 mimics, MEG3 siRNA (si-MEG3) and/or MEG3 overexpression plasmids (pc-MEG3) for 48 h. Cell migration, invasion, apoptosis, cell cycle distribution and the expression of E26 tansformation-specific-1 (ETS-1), phosphatase and tensin homologue (PTEN), E-cadherin, N-cadherin and ß-catenin were detected. RESULTS: MiR-377 mimics increased MEG3 expression and decreased the number of migrated and invaded U118 and U251 cells, without influence on apoptosis in both cell lines. Si-MEG3 transfection increased U118 cell migration and invasion and rescued miR-377 mimics-induced inhibitory in cell migration and invasion. Si-MEG3 decreased U118 cell apoptosis and induced G0/G1 cell cycle arrest, and pc-MEG3 increased U251 cell apoptosis via arresting cell cycle at G2/M phage. MiR-377 mimics and si-MEG3 increased the relative expression level of N-cadherin mRNA, and both si-MEG3 and pc-MEG3 increased E-cadherin in glioma cells. MiR-377 mimics increased ETS-1 mRNA in U118 cells, but decreased it in U251 cells. PTEN was increased by miR-377 mimics and si-MEG3 and decreased by pc-MEG3 in glioma cells. CONCLUSIONS: These results suggested the link interaction of MEG3 with miR-377 and PTEN, but not functioning as the competing endogenous RNA. MiR-377 mimics and MEG3 were tumor suppressors in glioma cells through regulating PTEN expression.

Real-time loop-mediated isothermal amplification for rapid detection of Enterocytozoon hepatopenaei.

BACKGROUND: Enterocytozoon hepatopenaei (EHP) is a newly emerged microsporidian parasite that causes retarded shrimp growth in many countries. But there are no effective approaches to control this disease to date. The EHP could be an immune risk factor for increased dissemination of other diseases. Further, EHP infection involves the absence of obvious clinical signs and it is difficult to identify the pathogen through visual examination, increasing the risk of disease dissemination. It is urgent and necessary to develop a specific, rapid and sensitive EHP-infected shrimp diagnostic method to detect this parasite. In the present study, we developed and evaluated a rapid real-time loop-mediated isothermal amplification (real-time LAMP) for detection of EHP. METHODS: A rapid and efficient real-time LAMP method for the detection of EHP has been developed. Newly emerged EHP pathogens in China were collected and used as the sample, and three sets of specificity and sensitivity primers were designed. Three other aquatic pathogens were used as templates to test the specificity of the real-time LAMP assay. Also, we compared the real-time LAMP with the conventional LAMP by the serial dilutions of EHP DNA and their amplification curves. Application of real-time LAMP was carried out with clinical samples. RESULTS: Positive products were amplified only from EHP, but not from other tested species, EHP was detected from the clinical samples, suggesting a high specificity of this method. The final results of this assay were available within less than 45 min, and the initial amplification curve was observed at about 6 min. We found that the amplification with an exponential of sixfold dilutions of EHP DNA demonstrated a specific positive signal by the real-time LAMP, but not for the LAMP amplicons from the visual inspection. The real-time LAMP amplification curves demonstrated a higher slope than the conventional LAMP. DISCUSSION: In this study, pathogen virulence impacts have been increased in aquaculture and continuous observation was predominantly focused on EHP. The present study confirmed that the real-time LAMP assay is a promising and convenient method for the rapid identification of EHP in less time and cost. Its application greatly aids in the detection, surveillance, and prevention of EHP.