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Microbial abundance and community composition in marine sediments have been widely explored. However, high‐resolution vertical changes of benthic microbial diversity and co‐occurrence patterns are poorly described. The ecological contributions of abundant and rare species in sediments also remain largely unknown. Here, by analysing microbial populations at 14 depth layers of 10 subseafloor sediment cores (water depth 1,250–3,530 m) obtained in the South China Sea, we provided the vertical profiles of microbial β‐diversity and co‐occurrence influenced by subcommunities of different abundance. These 134 sediment samples were clustered into four groups according to sediment depth (1–2, 6–10, 30–90 and 190–790 cm) with obvious shifts in microbial community compositions. The vertical succession of microorganisms was consistent with redox zonation and influenced by terrestrial inputs. Partitioning of vertical β‐diversity showed extremely high species replacement between deep layers and the surface layer, indicating selection‐induced loss of rare species and dispersal of dormant cells and spores. By contrast, for horizontal β‐diversity, richness of rare species became increasingly significant in deep sediments. Accompanying this β‐diversity profile were clear changes in the association pattern, with microorganisms being less connected in deeper sediment layers, probably reflecting reduced syntrophic interactions. Rare species accounted for an indispensable proportion in the co‐occurrence network, and tended to form complex “small worlds.” The rare subcommunity also responded differently to various environmental factors compared with the abundant subcommunity. Our findings expand current knowledge on vertical changes of marine benthic microbial diversity and their association patterns, emphasizing the potential roles of rare species.

Puccinia striiformis f. sp. tritici is a responsible pathogen for wheat stripe rust, which can be dispersed for a long distance by upper air flow. Understanding its origin and migration is vital to control the disease. We analyzed the population structures of the pathogen, which include 297 samples from Yunnan, Guizhou, Sichuan, Shaanxi, and Gansu in 2008 and 2011, by using the concatenated gene Cdc2-(Ef-1α)-Mapk1. From which, 18 vs 27 SNP loci, and 26 vs 23 haplotypes were tested in 2008 and 2011, respectively. The ancestral haplotype was detected in Yunnan in both years. Yunnan population had a higher mutation rate than Gansu population, which was 3.46e-03 vs 2.33e-03 in 2008, and 5.74e-03 vs 3.54e-03 in 2011, respectively. While the Gansu population had more recombination events than the Yunnan population, which was 3 vs 2 in 2008, and 5 vs 1 in 2011, respectively. The Gansu population had a higher genetic diversity than the Yunnan population, with the haplotype diversity of 0.7641 vs 0.7076 in 2008, and 0.8103 vs 0.4764 in 2011, respectively. There was a massive gene flow between Yunnan and Gansu, the Nm was 142.60 vs 9.47 in 2008 and 2011, respectively. The trajectories of upper air flow during wheat growing season from 2005 to 2013, directed mainly from Yunnan to Gansu. Based on above and previous researches regarding South Gansu as the crucial over-summering source of the stripe rust pathogen in China, Yunnan is suggested as the probable origin center for the pathogen in China, while Gansu is argued as the secondary center.

Background Maternal stress and nutrition during pregnancy are two of the most commonly studied factors in the context of fetal development. However, few investigators have considered the combined effects of stress and diet in pregnant women and their offspring. Aims To determine the impact of maternal stress on offspring birth weight, and the potential mediating role of maternal dietary patterns. Methods A total of 3542 pregnant women and their singleton live births were recruited from Shanghai Maternal-Child Pairs Cohort (Shanghai MCPC). Maternal stress was assessed using the Life Event Scale for Pregnant Women (LESPW) in early and late pregnancy. Food frequency questionnaire (FFQ) was performed to evaluate maternal diet in late pregnancy. Multiple linear regression was conducted to estimate the associations between maternal stress and child birth weight. Logistic regression was performed to calculate the association between maternal stress and small for gestational age (SGA)/large for gestational age (LGA). The bootstrap method was used to investigate the mediating effects of maternal dietary patterns. Results Maternal subjective events stress (β = 0.367) and total stress (β = 0.079) in early pregnancy, and positive objective life events stress (β = 0.275) in late pregnancy, were positively associated with birth weight. Maternal dietary pattern of “high-fat and sugar” mediated the associations between subjective events stress and total stress during early pregnancy and birth weight. An increased risk for LGA was observed among women exposed to relatively higher stress during early pregnancy (OR, 1.416; 95% CI, 1.035-1.937). No statistically significant associations were found between maternal stress during late pregnancy and child birth weight or SGA/LGA. Conclusions Maternal life events stress during early pregnancy significantly increases neonatal birth weight and risk for LGA, which may mediate by a “high-fat and sugar” dietary pattern. Limitations The lack of biological markers limits the explanation of the mechanism. The assessment of diet used food intake frequency to evaluate dietary characteristics, which may miss information of energy intake.

Metopolophium dirhodum (Walker) (Hemiptera: Aphididae) is one of the most common aphid pests of winter cereals. To facilitate accurate gene expression analyses with qRT-PCR assays, the expression stability of candidate reference genes under specific experimental conditions must be verified before they can be used to normalize target gene expression levels. In this study, 10 candidate reference genes in M. dirhodum were analyzed by qRT-PCR under various experimental conditions. Their expression stability was evaluated with delta Ct, BestKeeper, geNorm, and NormFinder methods, and the final stability ranking was determined with RefFinder. The results indicate that the most appropriate sets of internal controls were SDHB and RPL8 across geographic population; RPL8, Actin, and GAPDH across developmental stage; SDHB and NADH across body part; RPL8 and Actin across wing dimorphism and temperature; RPL4 and EF1A across starvation stress; AK and RPL4 across insecticide treatments; RPL8 and NADH across antibiotic treatments; RPL8, RPL4, Actin, and NADH across all samples. The results of this study provide useful insights for establishing a standardized qRT-PCR procedure for M. dirhodum and may be relevant for identifying appropriate reference genes for molecular analyses of related insects.

Dimethylsulfoniopropionate (DMSP) is an important marine osmolyte. Aphotic environments are only recently being considered as potential contributors to global DMSP production. Here, our Mariana Trench study reveals a typical seawater DMSP/dimethylsulfide (DMS) profile, with highest concentrations in the euphotic zone and decreased but consistent levels below. The genetic potential for bacterial DMSP synthesis via the dsyB gene and its transcription is greater in the deep ocean, and is highest in the sediment.s DMSP catabolic potential is present throughout the trench waters, but is less prominent below 8000 m, perhaps indicating a preference to store DMSP in the deep for stress protection. Deep ocean bacterial isolates show enhanced DMSP production under increased hydrostatic pressure. Furthermore, bacterial dsyB mutants are less tolerant of deep ocean pressures than wild-type strains. Thus, we propose a physiological function for DMSP in hydrostatic pressure protection, and that bacteria are key DMSP producers in deep seawater and sediment., Dimethylsulfoniopropionate (DMSP) is an osmolyte produced by marine microbes that plays an important role in nutrient cycling and atmospheric chemistry. Here the authors go to the Mariana Trench—the deepest point in the ocean—and find bacteria are key DMSP producers, and that DMSP has a role in protection against high pressure.

Purpose To explore the association between maternal physical activity (PA) and sleep quality during pregnancy, and the necessary PA level at different gestational stages to attain improved sleep quality. Methods A total of 2443 participants were recruited from the Shanghai Maternal-Child Pairs Cohort (Shanghai MCPC) study, who had completed questionnaires including the Pittsburgh Sleep Quality Index (PSQI) and the International Physical Activity Questionnaire (IPAQ) at gestational weeks (GW) of 12-16 and 32-36. PSQI scores and their seven components at the two GW were compared, as were PSQI scores at 12-16 and 32-36 GW and the increment in PSQI relative to PA. Regression analysis was conducted to assess the effect of PA and its change on the total PSQI score at different GW. Results The mean PSQI scores increased significantly during pregnancy, from 6.30 ± 3.01 at 12-16 GW to 7.23 ± 3.47 at 32-36 GW. Compared with women in low PA level, moderate levels of PA at both 12-16 GW and 32-36 GW were significantly reduced PSQI scores of 0.42 (95% CI:-0.68,-0.16) and 0.32 (95% CI:-0.63,-0.01), respectively. At 32-36 GW, high PA level also significantly decreased PSQI score, with a greater decline than moderate PA level. (AOR=-0.87,95% CI:-1.57,-0.18). PA increment from 12-16 to 32-36 weeks of pregnancy created a significant decline of 0.54 in PSQI scores. Conclusion The study revealed sleep quality was worse at the third trimester and moderate PA level had the potential for improvement of sleep quality both in the first and the third trimester. High PA level was also beneficial to improve sleep quality of pregnant women in the third trimester.

Background Recent research suggested that oxytocin might be associated with prenatal anxiety or depression. Also, our previous study indicated that resilience could buffer the effect of stress on prenatal anxiety and depression, which is a kind of psychological adjustment ability to stress in life. The effects of oxytocin and resilience are supposed to be similar but no study was carried out to find the association between them. Current evidence on the effect of oxytocin on prenatal mental health were with small sample size and equivocal results, especially with a lack of studies in Chinese pregnant women. Aims To investigate the relationship between oxytocin and resilience, and its role in regulating prenatal anxiety and depression among Chinese women. Methods The study was based on a sub-sample of the Shanghai Maternal-Child Pairs Cohort (Shanghai MCPC) by stratified random sampling. A total of 632 participants completed blood collection for serum oxytocin measurement and questionnaires pertaining to early pregnancy stress and resilience in the second trimester, which were assessed by the Life Event Scale for Pregnancy Women (LESPW) and the revised Resilience Scale for Adults (RSA), respectively. Prenatal anxiety and depression were assessed by Self-Rating Anxiety Scale (SAS) and the Center for Epidemiological Survey, Depression Scale (CES-D), respectively, at the third trimester. Correlation analysis and stratified linear regression analyses were conducted to understand the role of oxytocin in regulating prenatal anxiety and depression. Results The concentration of serum oxytocin in the second trimester ranged from 98.14 pg⋅ml−1 to 197.28 pg⋅ml−1, and there was no significant difference between different gestational age (r = 0.024, P = 0.579). The association between stress and prenatal anxiety was stronger in high oxytocin group than that in low oxytocin group, with B value of 3.338 and 2.638, respectively, after adjusting for residence, educational level, family income and parity. The same effect was found in the association between stress and prenatal depression, with B value of 2.360 and 2.060, respectively. However, it was not significantly associated with early pregnancy stress (Z = 1.933, P = 0.630), prenatal anxiety (Z = 1.387, P = 0.165), prenatal depression (Z = −0.027, P = 0.978), nor resilience (F = 1.282, P = 0.280). Limitations The existence of recall bias and restricted residential areas of the participants may limit the extrapolation of the study. Also, unsynchronized sampling timing of the oxytocin and psychological resilience measures is another limitation. Conclusions Serum oxytocin levels in the second trimester had an effect of modification between early pregnancy stress and prenatal anxiety and depression. Pregnant women with high oxytocin in the second trimester should receive more attention and further study is needed on the mechanism of oxytocin for the mental health of pregnant women.

Pyruvate kinase (PYK) is a speed-limited enzyme of glycolysis that catalyzes the formation of pyruvate, and plays an important role in acetyl-CoA synthesis. The acetyl-CoA is the precursor of sex pheromone biosynthesis in Helicoverpa armigera. However, the role of PYK in sex pheromone biosynthesis remains elusive. Here, PYK in H. armigera (HaPYK) was found to be highly expressed in the pheromone glands (PGs). The developmental expression profile of HaPYK was consistent with the fluctuation of sex pheromone release. Function analysis revealed that the knockdown of HaPYK led to a decrease in the levels of pyruvic acid and acetyl-CoA in PGs, which in turn caused a significant decrease in cis-11-hexadecenal (Z11-16: Ald) production, female capability to attract males, and mating frequency. Further study demonstrated that sugar feeding (5% sugar) increased the transcription and enzyme activity of HaPYK, thereby facilitating sex pheromone biosynthesis. Moreover, pheromone biosynthesis activating neuropeptide (PBAN) upregulated HaPYK activity through protein kinase C (PKC), as shown by PKC-specific inhibitor analysis. Altogether, our results revealed that PBAN activated HaPYK by Ca2+/PKC, thereby regulating the synthesis of pyruvate and subsequent acetyl-CoA, ensuring the supply of sex pheromone precursor, and finally facilitating sex pheromone biosynthesis and mating behavior.

Metagenomics and metatranscriptomics are powerful methods to uncover key micro-organisms and processes driving biogeochemical cycling in natural ecosystems. Databases dedicated to depicting biogeochemical pathways (for example, metabolism of dimethylsulfoniopropionate (DMSP), which is an abundant organosulfur compound) from metagenomic/metatranscriptomic data are rarely seen. Additionally, a recognized normalization model to estimate the relative abundance and environmental importance of pathways from metagenomic and metatranscriptomic data has not been organized to date. These limitations impact the ability to accurately relate key microbial-driven biogeochemical processes to differences in environmental conditions. Thus, an easy-to-use, specialized tool that infers and visually compares the potential for biogeochemical processes, including DMSP cycling, is urgently required. To solve these issues, we developed DiTing, a tool wrapper to infer and compare biogeochemical pathways among a set of given metagenomic or metatranscriptomic reads in one step, based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) and a manually created DMSP cycling gene database. Accurate and specific formulae for over 100 pathways were developed to calculate their relative abundance. Output reports detail the relative abundance of biogeochemical pathways in both text and graphical format. DiTing was applied to simulated metagenomic data and resulted in consistent genetic features of simulated benchmark genomic data. Subsequently, when applied to natural metagenomic and metatranscriptomic data from hydrothermal vents and the Tara Ocean project, the functional profiles predicted by DiTing were correlated with environmental condition changes. DiTing can now be confidently applied to wider metagenomic and metatranscriptomic datasets, and it is available at https://github.com/xuechunxu/DiTing.

A Gram-stain-negative, rod-shaped, non-motile, strictly aerobic strain, designated as MTEO17T, was isolated from a 1000 m deep seawater sample of the Mariana Trench. Growth was observed at 10–45 °C (optimum, 37 °C), in the presence of 0.0–12.0 % NaCl (w/v; optimum, 3.0 %) and at pH 6.0–10.0 (optimum, pH 7.0–8.0). Phylogenetic analysis, based on the 16S rRNA gene sequence, revealed that strain MTEO17T belonged to the genus Alcanivorax and showed the highest sequence similarity of 97.9 % to Alcanivorax nanhaiticus MCCC 1A05629T. The estimated average nucleotide identity and DNA–DNA hybridization values between strain MTEO17T and A. nanhaiticus MCCC 1A05629T were 78.98 and 23.80 %, respectively. The significant dominant fatty acids were C16 : 0, summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c) and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c). The polar lipids comprised two phosphatidylethanolamines, one phosphatidylglycerol, one unidentified phospholipid and four unidentified polar lipids. The DNA G+C content of strain MTEO17T was 57.5 %. On the basis of the polyphasic evidence, strain MTEO17T is proposed to represent a novel species of the genus Alcanivorax , for which the name Alcanivorax profundi sp. nov. is proposed. The type strain is MTEO17T (=KCTC 52694T=MCCC 1K03252T).

Dimethylsulfoniopropionate (DMSP) is one of Earth’s most abundant organosulfur molecules. Recently, many marine heterotrophic bacteria were shown to produce DMSP, but few studies have combined culture-dependent and independent techniques to study their abundance, distribution, diversity and activity in seawater or sediment environments. Here we investigate bacterial DMSP production potential in East China Sea (ECS) samples. Total DMSP (DMSPt) concentration in ECS seawater was highest in surface waters (SW) where phytoplankton were most abundant, and it decreased with depth to near bottom waters. However, the percentage of DMSPt mainly apportioned to bacteria increased from the surface to the near bottom water. The highest DMSP concentration was detected in ECS oxic surface sediment (OSS) where phytoplankton were not abundant. Bacteria with the genetic potential to produce DMSP and relevant biosynthesis gene transcripts were prominent in all ECS seawater and sediment samples. Their abundance also increased with depth and was highest in the OSS samples. Microbial enrichments for DMSP-producing bacteria from sediment and seawater identified many novel taxonomic groups of DMSP-producing bacteria. Different profiles of DMSP-producing bacteria existed between seawater and sediment samples and there are still novel DMSP-producing bacterial groups to be discovered in these environments. This study shows that heterotrophic bacteria significantly contribute to the marine DMSP pool and that their contribution increases with water depth and is highest in seabed surface sediment where DMSP catabolic potential is lowest. Furthermore, distinct bacterial groups likely produce DMSP in seawater and sediment samples, and many novel producing taxa exist, especially in the sediment.

Qianyu Yue,* Yanyan Xu,* Xiaoli Deng, Shenglan Wang, Jingman Qiu, Baojiang Qian, Yunhui Zhang Department of Pulmonary and Critical Care Medicine, The First People’s Hospital of Yunnan Province (The Affiliated Hospital of Kunming University of Science and Technology), Kunming, 650032, Yunnan, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yunhui Zhang; Xiaoli DengDepartment of Pulmonary and Critical Care Medicine, The First People’s Hospital of Yunnan Province (The Affiliated Hospital of Kunming University of Science and Technology), No. 157, Jinbi Road, Kunming, 650032, Yunnan, People’s Republic of ChinaTel +86 871-63638515Email Yueqianyu2020@163.com; xeygyc@163.comBackground: Circular RNA (circRNA) is a key regulator of cancer, and it has been proved to be involved in the regulation of cancer progression including non-small cell lung cancer (NSCLC). Circ-PITX1 was found to be a significantly upregulated circRNA in NSCLC, and its role and potential mechanism in NSCLC progression deserve further investigation.Methods: The expression levels of circ-PITX1, microRNA (miR)-1248 and cyclin D2 (CCND2) were examined by quantitative real-time PCR (qRT-PCR). Cell proliferation, apoptosis, cell cycle process, migration and invasion were determined using cell counting kit 8 (CCK8) assay, colony formation assay, flow cytometry, wound healing assay and transwell assay. Xenograft models were built to explore the role of circ-PITX1 in NSCLC tumor growth in vivo. The glycolysis and glutamine metabolism of cells were assessed by detecting the consumptions of glucose and glutamine, cell extracellular acidification rate (ECAR), and the productions of lactate, α-ketoglutaric acid (α-KG) and ATP. The protein levels of hexokinase 2 (HK-2), glutaminase 1 (GLS1) and CCND2 were tested by Western blot (WB) analysis. Dual-luciferase reporter assay and RIP assay were employed to verify the interaction between miR-1248 and circ-PITX1 or CCND2.Results: Circ-PITX1 was upregulated in NSCLC and its silencing could inhibit the proliferation, migration, invasion, cell cycle process, glycolysis, glutamine metabolism, and promote the apoptosis of NSCLC cells in vitro, as well as reduced tumor growth in vivo. In the terms of mechanism, we found that circ-PITX1 could act as a sponge of miR-1248, and miR-1248 could target CCND2. In addition, miR-1248 inhibitor reversed the inhibitory effect of circ-PITX1 knockdown on NSCLC progression. Similarly, CCND2 overexpression also reversed the suppressive effect of miR-1248 on NSCLC progression. Moreover, circ-PITX1 positively regulated CCND2 expression by sponging miR-1248.Conclusion: Circ-PITX1 served as a sponge of miR-1248 to promote NSCLC progression by upregulating CCND2.Keywords: non-small cell lung cancer, circ-PITX1, miR-1248, CCND2

Background Lung nodules are a diagnostic challenge. Current clinical management of lung nodule patients is inefficient and therefore causes patient misclassification, which increases healthcare expenses. However, a precise and robust lung nodule classifier to minimize discomfort for patients and healthcare costs is still lacking. The aim of the present protocol is to evaluate the effectiveness of using a liquid biopsy classifier to diagnose nodules compared to physician estimates and whether the classifier can reduce the number of unnecessary biopsies in benign cases. Methods A prospective cohort of 10,560 patients enrolled at 23 clinical centers in China with non-calcified pulmonary nodules, ranging from 0.5 to 3 cm in diameter, indicated by LDCT or CT will be included. After signed consent forms, the participants' pulmonary nodules will be assessed using three evaluation tools: (I) physician cancer probability estimates (II) validated lung nodule risk models, including Mayo Clinic and Veteran's Affairs models (III) ctDNA methylation classifier previously established. Each patient will undergo LDCT/CT follow-ups for 2 to 3 years and their information and one blood sample will be collected at baseline, 3, 6, 12, 24 and 36 months. The primary study outcomes will be the diagnostic accuracy of the methylation classifier in the cohort. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) will be used to compare the diagnostic value of each testing tool in differentiating benign and malignant pulmonary nodules. Discussion We are conducting an observational study to explore the accuracy of using a ctDNA methylation classifier for incidental lung nodules diagnosis. Trial registration Clinicaltrials.gov NCT03651986.

Bing Yuan,1,* Feng Liao,1,* Zhi-Zhou Shi,2 Yuan Ren,1 Xiao-Li Deng,1 Ting-Ting Yang,1 Deng-Yuan Li,1 Ru-Fang Li,1 Dan-Dan Pu,1 Yu-Jue Wang,1 Yan Tan,1 Zhen Yang,1 Yun-Hui Zhang1 1Department of Pulmonary and Critical Care Medicine, The First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology, Kunming, Yunnan, People’s Republic of China; 2Medical School, Kunming University of Science and Technology, Kunming, Yunnan, People’s Republic of China*These authors contributed equally to this work.Correspondence: Zhen YangDepartment of Pulmonary and Critical Care Medicine, The First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology, Kunming, Yunnan, People’s Republic of ChinaEmail 49387127@qq.comYun-Hui ZhangDepartment of Pulmonary and Critical Care Medicine, The First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology, Kunming, Yunnan, People’s Republic of ChinaEmail yunhuizhang3188@126.comObjective: Lung cancer is the first leading cause of cancer-related deaths both worldwide and in China and threatens human health and quality of life. New drugs and therapeutic methods are urgently needed. Our study evaluated the roles of dihydroartemisinin (DHA) in lung cancer and further explored its underlying mechanisms.Methods: CCK-8, colony formation and trypan blue exclusion assays were used to detect the cell viability, colony formation ability and cell death. qRT-PCR and Western blotting assays were applied to analyze the expressions of key molecules.Results: DHA inhibited the proliferation and colony formation abilities and enhanced the cell death and induced ferroptosis of lung NCI-H23 and XWLC-05 cancer cells. DHA reduced PRIM2 expression and silencing PRIM2 mimicked the inhibitory roles on proliferation and colony formation and promotive roles on cell death and ferroptosis of DHA in lung NCI-H23 and XWLC-05 cancer cells. We further found that DHA treatment and loss of PRIM2 reduced the GSH level and increased the cellular lipid ROS and mitochondrial MDA levels, and further downregulated the expressions of SLC7A11 and β-catenin in lung cancer cells, respectively. Exogenetic overexpression of PRIM2 recovered the inhibitory effects of DHA on proliferation and colony formation in lung NCI-H23 cancer cells, meanwhile loss of PRIM2 sensitizes NCI-H23 cells to DHA therapy. In vivo experiment further showed that DHA treatment significantly suppressed the tumor growth and downregulated PRIM2 and SLC7A11.Conclusion: Our study suggested that DHA inhibited the proliferation, colony formation and enhanced cell death and induced ferroptosis of lung cancer cells by inactivating PRIM2/SLC7A11 axis. Loss of PRIM2 induced ferroptosis might developed to be a novel therapeutic method in lung cancer therapy.Keywords: dihydroartemisinin, lung cancer, proliferation, ferroptosis, PRIM2, SLC7A11

Vilaprisan is a novel selective progesterone receptor modulator for the long‐term treatment of uterine fibroids and endometriosis. This study investigated the pharmacokinetics, safety, and tolerability of vilaprisan in healthy Chinese postmenopausal women. Twelve participants received multiple doses of vilaprisan once daily over 14 days as a 2‐mg tablet. Plasma vilaprisan concentrations were determined using liquid chromatography–tandem mass spectrometry. The main pharmacokinetic parameters of vilaprisan were assessed with noncompartmental analysis, including maximum observed concentration (Cmax), systemic exposure (area under the plasma concentration–time curve), time to reach Cmax and terminal half‐life. Safety assessments include the documentation of adverse events, measurement of clinical/anthropometric parameters and vital signs, electrocardiogram, and physical and gynecologic examination. The participants had a mean age of 53.3 (± 4.2) years and a body mass index of 23.8 ± 2.8 kg/m2. Median time to reach Cmax was 1.5 hours after both single and multiple vilaprisan administration. Mean Cmax values obtained after multiple dosing (23.3 μg/L [standard deviation (SD) = 6.73]) were 1.92‐fold (SD = 0.554) higher compared to single dosing (12.5 μg/L [SD = 3.04]). Mean area under the plasma concentration–time curve in the dosing interval increased with an accumulation factor of 2.98 (SD = 0.767) between single (91.3 μg · h/L [SD = 20.4]) and multiple dosing (276 μg · h/L [SD = 109]). The mean terminal half‐life of vilaprisan was 44.5 hours (SD = 10.3) after multiple dosing. Mild to moderate adverse events were observed similar to previous studies. Overall, daily oral administration of the therapeutic dose of 2 mg of vilaprisan over 14 days was safe and well tolerated by all participants.

Recently, several miRNAs have been revealed to play critical roles in oncogenesis and tumor progression of many cancers. Thioredoxin-1 (Trx-1) binding protein-2 (TBP-2) is an internal inhibitor of Trx-1, which plays the role in regulating oxidative stress, inhibiting cell growth, and promoting apoptosis. The expression of TBP-2 is usually decreased in cancer tissues. However, whether the miRNAs regulate the TBP-2 expression in lung cancer is still unclear. In this study, we examined the levels of TBP-2, miR-93, miR-373, and miR-17-5p in lung cancer tissues and their adjacent normal lung tissues of 36 patients. We found that the expressions of miR-93, miR-373, and miR-17-5p were higher, whereas the expression of TBP-2 mRNA and protein was significantly lower in lung cancer tissues compared with adjacent normal lung tissues. After the three miRNA mimics were transfected in the lung cancer cells, NCI-H460, the level of TBP-2 mRNA and TBP-2 protein was decreased. Then, the anti-cancer drug 5-fluorouracil was used to stimulate the NCI-H460 cells; the mRNA levels of miR-93, miR-373, and miR-17-5p were decreased, and the level of TBP-2 mRNA and protein was increased. Collectively, the above results suggest that miR-93, miR-373, and miR-17-5p negatively regulate the TBP-2 expression in lung cancer. This study may provide therapeutic targets with lung cancer.

Insect Odorant-Binding Proteins (OBPs) play crucial roles in the discrimination, binding and transportation of odorants. Herein, the full-length cDNA sequence of Minus-C OBP1 (MaltOBP1) from the Japanese pine sawyer beetle, Monochamus alternatus, was cloned by 3′ and 5′ RACE-PCR and analyzed. The results showed that MaltOBP1 contains a 435 bp open reading frame (ORF) that encodes 144 amino acids, including a 21-amino acid signal peptide at the N-terminus. The matured MaltOBP1 protein possesses a predicted molecular weight of about 14 kDa and consists of six α-helices, creating an open binding pocket, and two disulfide bridges. Immunoblotting results showed that MaltOBP1 was most highly expressed in antennae in both sexes, followed by wings and legs. Fluorescence assays demonstrated that MaltOBP1 protein exhibited high binding affinity with (R)-(+)-α-pinene, (−)-β-pinene, trans-caryophyllene, (R)-(+)-limonene and (–)-verbenone, which are the main volatile compounds of the pine tree. Our combined results suggest that MaltOBP1 plays a role in host seeking behavior in M. alternatus.

The microbial cycling of dimethylsulfoniopropionate (DMSP) and its gaseous catabolites dimethylsulfide (DMS) and methanethiol (MeSH) are important processes in the global sulfur cycle, marine microbial food webs, signaling pathways, atmospheric chemistry, and potentially climate regulation. Many functional genes have been identified and used to study the genetic potential of microbes to produce and catabolize these organosulfur compounds in different marine environments. Here, we sampled seawater, marine sediment and hydrothermal sediment, and polymetallic sulfide in the eastern Chinese marginal seas and analyzed their microbial communities for the genetic potential to cycle DMSP, DMS, and MeSH using metagenomics. DMSP was abundant in all sediment samples, but was fivefold less prominent in those from hydrothermal samples. Indeed, Yellow Sea (YS) sediment samples had DMSP concentrations two orders of magnitude higher than in surface water samples. Bacterial genetic potential to synthesize DMSP (mainly in Rhodobacteraceae bacteria) was far higher than for phytoplankton in all samples, but particularly in the sediment where no algal DMSP synthesis genes were detected. Thus, we propose bacteria as important DMSP producers in these marine sediments. DMSP catabolic pathways mediated by the DMSP lyase DddP (prominent in Pseudomonas and Mesorhizobium bacteria) and DMSP demethylase DmdA enzymes (prominent in Rhodobacteraceae bacteria) and MddA-mediated MeSH S-methylation were very abundant in Bohai Sea and Yellow Sea sediments (BYSS) samples. In contrast, the genetic potential for DMSP degradation was very low in the hydrothermal sediment samples—dddP was the only catabolic gene detected and in only one sample. However, the potential for DMS production from MeSH (mddA) and DMS oxidation (dmoA and ddhA) was relatively abundant. This metagenomics study does not provide conclusive evidence for DMSP cycling; however, it does highlight the potential importance of bacteria in the synthesis and catabolism of DMSP and related compounds in diverse sediment environments.

Peroxisome proliferator-activated receptors (PPARs), members of the nuclear hormone receptor family, attract wide attention as promising therapeutic targets for the treatment of multiple diseases, and their target selective ligands were also intensively developed for pharmacological agents such as the approved drugs fibrates and thiazolidinediones (TZDs). Despite their potent pharmacological activities, PPARs are reported to be involved in agent- and pollutant-induced multiple organ toxicity or protective effects against toxicity. A better understanding of the protective and the detrimental role of PPARs will help to preserve efficacy of the PPAR modulators but diminish adverse effects. The present review summarizes and critiques current findings related to PPAR-mediated types of toxicity and protective effects against toxicity for a systematic understanding of PPARs in toxicology and applied pharmacology.

Molecular mechanisms underlying wing evolution and development have been a point of scientific inquiry for decades. Phloem-feeding aphids are one of the most devastating global insect pests, where dispersal of winged morphs lead to annual movements, migrations, and range expansions. Aphids show a polyphenic wing dimorphism trait, and offer a model to study the role of environment in determining morphological plasticity of a single genotype. Despite recent progresses in the genetic understanding of wing polyphenism, the influence of environmental cues remains unclear. To investigate the involvement of miRNAs in wing development, we sequenced small RNA libraries of the English grain aphid, Sitobion avenae (F.), across six different developmental stages. As a result, we identified 113 conserved and 193 S. avenae-specific miRNAs. Gene Ontology and KEGG pathway analyses of putative target mRNAs for the six differentially expressed miRNAs are enriched for wing development processes. Dietary uptake of miR-263a, miR-316, and miR-184a agomirs and antagomirs led to significantly higher mortality (>70%) and a lower proportion of winged morphs (

Objective: Metabolomics may unravel global metabolic changes in response to environmental exposures and identify important biological pathways involved in the pathophysiology of childhood obesity. Phthalate has been considered an obesogen and contributing to overweight and obesity in children. The purpose of this study is to evaluate changes in urine metabolites in response to the environmental phthalate exposure among overweight or obese children, and to investigate the metabolic mechanisms involved in the obesogenic effect of phthalate on children at puberty. Methods: Within the national Puberty Timing and Health Effects in Chinese Children (PTHEC) study, 69 overweight/obese children and 80 normal weight children were selected into the current study according to their puberty timing and WGOC (The Working Group for obesity in China) references. Urinary concentrations of six phthalate monoesters (MMP, MEP, MnBP, MEHP, MEOHP and MEHHP) were measured using API 2000 electrospray triple quadrupole mass spectrometer (ESIMS/MS). Metabolomic profiling of spot urine was performed using gas chromatography-mass spectrometry. Differentially expressed urinary metabolites associated with phthalate monoesters exposure were examined using orthogonal partial least square-discriminant analysis and multiple linear regression models. In addition, the candidate metabolites were regressed to obesity indices with multiple linear regression models and logistic regression models in all subjects. Results: Compared with normal weight children, higher levels of MnBP were detected in urinary samples of children with overweight and obesity. After adjusting for confounders including chronological age, gender, puberty onset, daily energy intake and physical activity and socio-economic level, positive association remained between urinary MnBP concentration and childhood overweight/obesity [OR = 1.586, 95% CI:1.043,2.412]. We observed elevated MnBP concentration was significantly correlated with increased levels of monostearin, 1-monopalmitin, stearic acid, itaconic acid, glycerol 3-phosphate, 5-methoxytryptamine, kyotorphin, 1-methylhydantoin, d-alanyl-d-alanine, pyrrole-2-carboxylic acid, 3,4-Dihydroxyphenylglycol, and butyraldehyde. Meanwhile, increased MnBP concentration was also significantly correlated with decreased levels of lactate, glucose 6-phosphate, d-fructose 6-phosphate, palmitic acid, 4-acetamidobutyric acid, l-glutamic acid, n-acetyl-l-phenylalanine, iminodiacetic acid, hydroxyproline, pipecolinic acid, l-ornithine, n-acetyl-l-glutamic acid, guanosine, cytosin, and (s)-mandelic acid in the normal weight subjects. The observations indicated that MnBP exposure was related to global urine metabolic abnormalities characterized by disrupting arginine and proline metabolism and increasing oxidative stress and fatty acid reesterification. Among the metabolic markers related to MnBP exposure, 1-methylhydantoin, pyrrole-2-carboxylic acid and monostearin were found to be positively correlated with obesity indices, while hydroxyproline, l-ornithine, and lactate were negatively associated with overweight/obesity in children. Conclusions: Our results suggested that the disrupted arginine and proline metabolism associated with phthalate exposure might contribute to the development of overweight and obesity in school-age children, providing insights into the pathophysiological changes and molecular mechanisms involved in childhood obesity. Keywords: Phthalate, Childhood overweight/obesity, Metabolic profiles, Arginine and proline metabolism

Outbreaks of avian influenza virus continue to pose threats to human health. Animal models such as the mouse, ferret, and macaque are used to understand the pathogenesis of avian influenza virus infection in humans. We previously reported that the tree shrew (Tupaia belangeri, family Tupaiidae), which is regarded as a “low-level primate”, has α2,3- and α2,6-linked sialic acid receptor distributions similar to those of humans and is potentially a useful mammalian model for studying mild human influenza (H1N1) virus infection. In this study, we used the tree shrew experimental model to investigate the pathogenesis of avian influenza A (H9N2) virus infection and the effect of the E627K mutation in the PB2 gene, an adaptation to mammalian hosts. Evidence of disease, virus titers in the upper and lower respiratory tract, histopathology and induction of proinflammatory cytokines are described. We also established ex vivo culture models of tree shrew respiratory tissues to study the tropism and replication of the H9N2 virus. Our results demonstrated that the tree shrew is a viable new in vivo experimental model for avian influenza research that provides results comparable to those observed in ferrets. The disease spectrum and pathogenesis in tree shrews correlate well with what is observed in humans.

Previous studies have linked prenatal polybrominated diphenyl ether (PBDE) exposure to adverse birth outcomes. Reductions in placental size might mediate those adverse effects. We examined whether umbilical cord blood PBDEs concentrations were associated with placental size at birth. A total of 121 mother-newborn pairs residing in Wenzhou were enrolled in this study. Concentrations of 19 PBDEs in umbilical cord blood were determined by gas chromatography tandem mass spectrometry (GC-MS). Placental length, breadth and surface area was measured by medical professionals after delivery. BDE-206, 207, 208, 209 and the total of 19 PBDE congeners concentrations were negatively associated with placental length, breadth and surface area. Our results suggest that prenatal PBDEs exposure may suppress placental growth, resulting in reduced placental size.

Lepidoptera sex pheromone biosynthesis is regulated by pheromone biosynthesis activating neuropeptide (PBAN). PBAN regulates not only female sex pheromone biosynthesis but also male sex pheromone biosynthesis. Previous research has confirmed that PBAN regulates sex pheromone biosynthesis using Ca2+ as a secondary messenger in all examined species to date. However, the downstream signal of Ca2+ has remained elusive. In the present study, calcineurin A (CNA), a downstream signal of Ca2+ , was discovered in Helicoverpa armigera male hairpencil and named HaCNA. Sequence analysis demonstrated that the open reading frame of HaCNA contains 1488 nucleotides encoding 495 amino acid residues. A homology search revealed that HaCNA shares a high amino acid identity with the CNA of other insects. Developmental and spatial expression analyses revealed that the mRNA levels of HaCNA peaked at 24 h after emergence and that HaCNA expression was ubiquitous in all examined tissues. Activity analysis revealed that PBAN activates HaCNA, and a Ca2+ inhibitor, Lacl3 , attenuated the effect of PBAN by decreasing HaCNA activity. Pharmacological inhibitor and RNA interference-mediated knockdown assays revealed that both activity inhibition and decreased mRNA levels of HaCNA led to a significant decrease in the production of the male sex pheromone components [octadecanol and (Z)-11 hexadecanol)] and in the efficacy of female mating acceptance. Our results demonstrate that HaCNA acts as downstream signal of PBAN/Ca2+ and plays an important role in PBAN-induced male sex pheromone biosynthesis and female mating acceptance.

Insect pests are primarily controlled by insecticides. However, the sensitivity decreases and insecticide resistance is problematic for the effective management of agriculturally important insects, including Metopolophium dirhodum, which is an aphid that commonly feeds on cereals. The insecticide sensitivity status and potential resistance of M. dirhodum field populations remain relatively unknown. In this study, the susceptibility of 19 M. dirhodum populations from seven provinces in Northern China to neonicotinoids, pyrethroids, organophosphates, and a macrolide (abamectin) was determined in 2017–2019. The results indicated that two populations were highly resistant to thiamethoxam, with a relative resistance ratio (RLR) of 134.03 and 103.03, whereas one population was highly resistant to beta-cypermethrin (RLR of 121.42). On the basis of the RLR, the tested M. dirhodum populations ranging from susceptible to showing moderate levels of resistance to imidacloprid (RLR of 1.50 to 57.29), omethoate (RLR of 1.07 to 18.73), and abamectin (RLR of 1.10 to 25.89), but they were ranging from susceptible to showing tolerance or low levels of resistance to bifenthrin (RLR of 1.14 to 6.02) and chlorpyrifos (RLR of 1.11 to 7.59). Furthermore, a pair-wise correlation analysis revealed a significant correlation between the median lethal concentrations (LC50) for beta-cypermethrin and thiamethoxam, reflecting the cross-resistance between these two insecticides. The data obtained in our study provide timely information about aphid insecticide sensitivity, which may be used to delay the evolution of M. dirhodum insecticide resistance in Northern China.

Triptolide (TP), a primary bioactive ingredient isolated from the traditional Chinese herbal medicine Tripterygium wilfordii Hook. F. (TWHF), has attracted great interest for its therapeutic biological activities in inflammation and autoimmune disease. However, its clinical use is limited by severe testicular toxicity, and the underlying mechanism has not been elucidated. Our preliminary evidence demonstrated that TP disrupted glucose metabolism and caused testicular toxicity. During spermatogenesis, Sertoli cells (SCs) provide lactate as an energy source to germ cells by glycolysis. The transcription factors GATA-binding protein 4 (GATA4) and specificity protein 1 (Sp1) can regulate glycolysis. Based on this evidence, we speculate that TP causes abnormal glycolysis in SCs by influencing the expression of the transcription factors GATA4 and Sp1. The mechanism of TP-induced testicular toxicity was investigated in vitro and in vivo. The data indicated that TP decreased glucose consumption, lactate production, and the mRNA levels of glycolysis-related transporters and enzymes. TP also downregulated the protein expression of the transcription factors GATA4 and Sp1, as well as the glycolytic enzyme phosphofructokinase platelet (PFKP). Phosphorylated GATA4 and nuclear GATA4 protein levels were reduced in a dose- and time-dependent manner after TP incubation. Similar effects were observed in shGata4-treated TM4 cells and BALB/c mice administered 0.4 mg/kg TP for 28 days, and glycolysis was also inhibited. Gata4 knockdown downregulated Sp1 and PFKP expression. Furthermore, the Sp1 inhibitor plicamycin inhibited PFKP protein levels in TM4 cells. In conclusion, TP inhibited GATA4-mediated glycolysis by suppressing Sp1-dependent PFKP expression in SCs and caused testicular toxicity.

The aim of the study was to determine whether applying an assistance force to the pelvis and legs during treadmill training can improve walking function in children with cerebral palsy.Twenty-three children with cerebral palsy were randomly assigned to the robotic or treadmill only group. For participants who were assigned to the robotic group, a controlled force was applied to the pelvis and legs during treadmill walking. For participants who were assigned to the treadmill only group, manual assistance was provided as needed. Each participant trained 3 times/wk for 6 wks. Outcome measures included walking speed, 6-min walking distance, and clinical assessment of motor function, which were evaluated before, after training, and 8 wks after the end of training, and were compared between two groups.Significant increases in walking speed and 6-min walking distance were observed after robotic training (P = 0.03), but no significant change was observed after treadmill training only. A greater increase in 6-min walking distance was observed after robotic training than that after treadmill only training (P = 0.01).Applying a controlled force to the pelvis and legs, for facilitating weight-shift and leg swing, respectively, during treadmill training may improve walking speed and endurance in children with cerebral palsy.Complete the self-assessment activity and evaluation online at http://www.physiatry.org/JournalCME CME OBJECTIVES: Upon completion of this article, the reader should be able to: (1) discuss the importance of physical activity at the participation level (sports programs) for children with cerebral palsy; (2) contrast the changes in walking ability and endurance for children in GMFCS level I, II and III following sports programs; and (3) identify the impact of higher frequency of sports program attendance over time on walking ability.Advanced ACCREDITATION: The Association of Academic Physiatrists is accredited by the Accreditation Council for Continuing Medical Education to provide continuing medical education for physicians.The Association of Academic Physiatrists designates this Journal-based CME activity for a maximum of 0.5 AMA PRA Category 1 Credit(s)™. Physicians should only claim credit commensurate with the extent of their participation in the activity.

The early postnatal period is the most dynamic and vulnerable stage in the assembly of intestinal microbiota. Antibiotics are commonly prescribed to newborn preterm babies and are frequently used for a prolonged duration in China. We hypothesized that the prolonged antibiotic therapy would affect the early development of intestinal microbiota and their metabolites. To test this hypothesis, we analyzed the stool microbiota and metabolites in 36 preterm babies with or without antibiotic treatment. These babies were divided into three groups, including two groups treated with the combination of penicillin and moxalactam or piperacillin-tazobactam for 7 days, and the other group was free of antibiotics. Compared to the antibiotic-free group, both antibiotic-treated groups had distinct gut microbial communities and metabolites, including a reduction of bacterial diversity and an enrichment of harmful bacteria such as Streptococcus and Pseudomonas. In addition, there was a significant difference in the composition of gut microbiota and their metabolites between the two antibiotic-treated groups, where the piperacillin-tazobactam treatment group showed an overgrowth of Enterococcus. These findings suggest that prolonged antibiotic therapy affects the early development of gut microbiota in preterm infants, which should be considered when prescribing antibiotics for this population.

Background Currently, Staphylococcus aureus is one of the most important pathogens worldwide, especially for methicillin-resistant S. aureus (MRSA) infection. However, few reports referred to patients’ MRSA infections in Yunnan province, southwest China. Methods In this study, we selected representative MRSA strains from patients’ systemic surveillance in Yunnan province of China, performed the genomic sequencing and compared their features, together with some food derived strains. Results Among sixty selective isolates, forty strains were isolated from patients, and twenty isolated from food. Among the patients’ strains, sixteen were recognized as community-acquired (CA), compared with 24 for hospital-acquired (HA). ST6-t701, ST59-t437 and ST239-t030 were the three major genotype profiles. ST6-t701 was predominated in food strains, while ST59-t437 and ST239-t030 were the primary clones in patients. The clinical features between CA and HA-MRSA of patients were statistical different. Compared the antibiotic resistant results between patients and food indicated that higher antibiotic resistant rates were found in patients’ strains. Totally, the average genome sizes of 60 isolates were 2.79 ± 0.05 Mbp, with GC content 33% and 84.50 ± 0.20% of coding rate. The core genomes of these isolates were 1593 genes. Phylogenetic analysis based on pan-genome and SNP of strains showed that five clustering groups were generated. Clustering ST239-t030 contained all the HA-MRSA cases in this study; clustering ST6-t701 referred to food and CA-MRSA infections in community; clustering ST59-t437 showed the heterogeneity for provoking different clinical diseases in both community and hospital. Phylogenetic tree, incorporating 24 isolates from different regions, indicated ST239-t030 strains in this study were more closely related to T0131 isolate from Tianjin, China, belonged to ‘Turkish clade’ from Eastern Europe; two groups of ST59-t437 clones of MRSA in Yunnan province were generated, belonged to the ‘Asian-Pacific’ clone (AP) and ‘Taiwan’ clone (TW) respectively. Conclusions ST239-t030, ST59-t437 and ST6-t701 were the three major MRSA clones in Yunnan province of China. ST239-t030 clonal Yunnan isolates demonstrated the local endemic of clone establishment for a number of years, whereas ST59-t437 strains revealed the multi-origins of this clone. In general, genomic study on epidemic clones of MRSA in southwest China provided the features and evolution of this pathogen.

Microbial production and catabolism of dimethylsulfoniopropionate (DMSP), generating the climatically active gases dimethyl sulfide (DMS) and methanethiol (MeSH), have key roles in global carbon and sulfur cycling, chemotaxis, and atmospheric chemistry. Microorganisms in the sea surface microlayer (SML), the interface between seawater and atmosphere, likely play an important role in the generation of DMS and MeSH and their exchange to the atmosphere, but little is known about these SML microorganisms. Here, we investigated the differences between bacterial community structure and the distribution and transcription profiles of the key bacterial DMSP synthesis (dsyB and mmtN) and catabolic (dmdA and dddP) genes in East China Sea SML and subsurface seawater (SSW) samples. Per equivalent volume, bacteria were far more abundant (~ 7.5-fold) in SML than SSW, as were those genera predicted to produce DMSP. Indeed, dsyB (~ 7-fold) and mmtN (~ 4-fold), robust reporters for bacterial DMSP production, were also far more abundant in SML than SSW. In addition, the SML had higher dsyB transcripts (~ 3-fold) than SSW samples, which may contribute to the significantly higher DMSP level observed in SML compared with SSW. Furthermore, the abundance of bacteria with dmdA and their transcription were higher in SML than SSW samples. Bacteria with dddP and transcripts were also prominent, but less than dmdA and presented at similar levels in both layers. These data indicate that the SML might be an important hotspot for bacterial DMSP production as well as generating the climatically active gases DMS and MeSH, a portion of which are likely transferred to the atmosphere.

This meta-analysis evaluates the difference in deep inspiration breath hold (DIBH) versus free breathing (FB) for patients receiving postoperative radiotherapy for left breast cancer and provides a useful reference for clinical practice. The relevant controlled trials of DIBH versus FB in postoperative radiotherapy for left-side breast cancer were retrieved from the databases of PubMed, Science Direct, Cochrane Library, and Web of Science databases. The principal outcome of interest was heart dose, left anterior descending coronary artery (LADCA) dose, and left lung dose and target coverage. We calculated summary standardized mean difference (SMD) and 95% confidence intervals (CI). The meta-analysis was performed using RevMan 5.3 software. The analysis included 1019 patients from 12 observational studies, of which 576 cases were in the DIBH group and 443 cases in the FB group. Compared with the FB group, the DIBH group can have lower heart dose, left anterior descending coronary artery (LADCA) dose, and left lung dose more effectively, and the difference was statistically significant (heart dose, SMD = − 1.36, 95% CI − 1.64 ~ − 1.09, P

MomL is a marine-derived quorum-quenching (QQ) lactonase which can degrade various N-acyl homoserine lactones (AHLs). Intentional modification of MomL may lead to a highly efficient QQ enzyme with broad application potential. In this study, we used a rapid and efficient method combining error-prone polymerase chain reaction (epPCR), high-throughput screening and site-directed mutagenesis to identify highly active MomL mutants. In this way, we obtained two candidate mutants, MomLI144V and MomLV149A. These two mutants exhibited enhanced activities and blocked the production of pathogenic factors of Pectobacterium carotovorum subsp. carotovorum (Pcc). Besides, seven amino acids which are vital for MomL enzyme activity were identified. Substitutions of these amino acids (E238G/K205E/L254R) in MomL led to almost complete loss of its QQ activity. We then tested the effect of MomL and its mutants on Pcc-infected Chinese cabbage. The results indicated that MomL and its mutants (MomLL254R, MomLI144V, MomLV149A) significantly decreased the pathogenicity of Pcc. This study provides an efficient method for QQ enzyme modification and gives us new clues for further investigation on the catalytic mechanism of QQ lactonase.

Triptolide (TP), a principal bioactive component extracted from traditional Chinese medicine Tripterygium wilfordii Hook. F. (TWHF), has attracted wide attention of its therapeutic effects on inflammation and autoimmune diseases. However, the therapeutic application of TP is hindered by severe cardiomyocyte toxicity and narrow therapeutic window. We previously identified that the p53 was an indispensable contributor in TP-induced myocardial injury. p53 has an inhibitory effect on IKKβ-NF-κB pathway that regulates glucose transporters (GLUT) expression. Based on these evidences, we speculate that p53 mediates TP-disturbed glucose uptake by blocking IKKβ-NF-κB signaling. This study focused on the effect of TP on cardiac glucose uptake and the role of p53 in glucose metabolism in cardiomyocytes, and p53 −/− mice. TP treatment depressed glucose consumption and ATP production resulting in myocardial damage. Incubation with ATP (5 mM) remarkably decreased the cellular damage. Immunoblotting and immunofluorescence identified that TP suppressed glucose uptake by restricting IKKβ-NF-κB signaling activation, GLUT1 and GLUT4 expression. p53 inhibition alleviated the cell damage and the compromise of glucose uptake. Mechanistically, p53 antagonist PFTα abolished TP-induced the inhibition of IKKβ, IκBα phosphorylation, p65 nuclear translocation, and GLUT1, GLUT4 expression. Consistently, in acute heart injury models, p53 deficiency upregulated IKKβ-NF-κB activation and GLUT1, GLUT4 protein levels which was also indicated as amelioration of heart histological injury after 1.2 mg kg−1 TP administration. The present findings indicate that TP-induced p53 overactivation suppresses glucose uptake by inhibiting IKKβ–NF-κB pathway and downregulating NF-κB-dependent GLUT1 and GLUT4 expression.

Isovaleryl-CoA dehydrogenase (IVD), a member of the acyl-CoA dehydrogenase (ACAD) family, is a key enzyme catalyzing the conversion of isovaleryl-CoA to β-methylcrotonyl-CoA in the third reaction of the leucine catabolism pathway and simultaneously transferring electrons to the electron-transferring flavoprotein (ETF) for ATP synthesis. We previously identified the ETF ortholog in rice blast fungus Magnaporthe oryzae (MoETF) and showed that MoETF was essential for the fungal growth, conidiation and pathogenicity. To further investigate the biological function of electron-transferring proteins and clarify the role of leucine catabolism in growth and pathogenesis, we characterized MoIVD (M. oryzae isovaleryl-CoA dehydrogenase). MoIvd is highly conserved in fungi and its expression was highly induced by leucine. The Δmoivd mutants showed reduced growth, decreased conidiation and compromised pathogenicity, while the conidial germination and appressorial formation appeared normal. Consistent with a block in leucine degradation, the Δmoivd mutants accumulated isovaleric acid, and grew more slowly, fully lacked pigmentation and completely failed to produce conidia on leucine-rich medium. These defects were largely rescued by raising the extracellular pH, suggesting that the accumulation of isovaleric acid contributes to the growth and conidiation defects. However, the reduced virulence of the mutants was probably due to their inability to overcome oxidative stress, since a large amount of ROS (reactive oxygen species) accumulated in infected host cell. In addition, MoIvd is localized to mitochondria and interacted with its electron receptor MoEtfb, the β subunit of MoEtf. Taken together, our results suggest that MoIVD functions in leucine catabolism and is required for the vegetative growth, conidiation and full virulence of M. oryzae, providing the first evidence for IVD-mediated leucine catabolism in the development and pathogenesis of plant fungal pathogens.

To determine whether repeat exposure to force perturbations during treadmill training can induce long-term retention of improved step length and overall improvements in locomotor function in persons with spinal cord injury.Fourteen patients with spinal cord injury were recruited and randomly assigned to swing resistance or swing assistance training groups. A controlled swing resistance or assistance force, for resistance or assistance training groups, respectively, was applied to both legs through a cable-driven robotic system during treadmill training. Each participant trained 3 times per week for 6 weeks. Step length, walking speed, 6-minute walking distance, and other clinical assessments were evaluated before and after 6 weeks of training and 8 weeks after the end of training.A significant increase in step length was observed after 6 weeks of resistance training (P = 0.04). Step length tended to increase after assistance treadmill training, but the change was not significant (P = 0.18). The changes in step length and functional gains had no significant difference between 2 groups.Repeat exposure to swing resistance during treadmill training may induce a prolonged retention of increased step length, although it remains unclear whether swing resistance versus assistance is more effective in inducing increased step length.

This Letter describes an efficient ring-closing metathesis approach to 2-chloro-4-amino-pyrimido[4,5-c]azepines and 2-chloro-4-amino-pyrimido[4,5-c]oxepines. These chlorides were applied to the synthesis of several potent γ-secretase modulators (GSMs).

Background Influenza C virus causes mild respiratory diseases in humans. Previous studies suggested that the predominant hemagglutinin-esterase gene lineage circulating in children might be selected among the adult population, yet the prevalence of influenza C virus in adults has not been described. Objectives To evaluate the frequency of influenza C virus infection in adults. Study design We performed hemagglutination inhibition assays of serum samples collected at periodic occupational medical checkups from employees of a hospital. A total of 679 serum samples were collected from 57 subjects who participated in biannual medical checkups between 2011 and 2016 as part of a longitudinal series. Titers of antibodies against the C/Kanagawa and C/Sao Paulo lineage viruses were detected. Results Ten serum sample pairs from among the 57 subjects showed at least a four-fold increase in influenza C antibody titers. Samples from three subjects exhibited antibody titer increases for both the C/Kanagawa and C/Sao Paulo lineages, four subjects showed an increased titer against the C/Sao Paulo lineage, and three subjects showed an increased titer against the C/Kanagawa lineage. Half of the antibody titer increases for the C/Kanagawa lineage were detected in May 2014, while the increases for the C/Sao Paulo lineage were detected from 2011 to 2016. Conclusion The 5-year influenza C virus infection rate was estimated at 17.5 %. There were antibodies that cross-reacted with the C/Sao Paulo and C/Kanagawa lineages. The results suggest that C/Sao Paulo was the main lineage in the adult population of this area, with cocirculation of the C/Kanagawa lineage.

Enterovirus A71 (EV71) remains the most common causative agent of hand, foot, and mouth disease (HFMD), and the neurological complications induced by EV71 are usually the leading cause of death in children with HFMD. However, the mechanism of nervous system changes caused by EV71 infection is still unclear. Therefore, in the current study, EV71 was inoculated into the human neuroblastoma cell line SH-SY5Y and subsequent transcriptome sequencing was used to examine the alterations of the transcriptome in infected SH-SY5Y cells. It is expected to determine the underlying mechanism of neurological diseases in response to EV71 infection. As a result, a total of 82,406,974, 112,410,808 and 87,780,371 clean reads were found in the control, EV71-12 h and EV71-24 h groups, respectively. Moreover, 160 and 745 differentially expressed genes were identified in the EV71-12 h and EV71-24 h groups, respectively, as compared to the control group. Next, to further explore the pathogenic mechanism triggered by EV71 infection, we mainly focused on the common differentially expressed genes at different time points of EV71 infection. And it was discovered that there were 95 common differentially expressed genes, which were used to conduct GO and pathway analysis. GO enrichment analysis demarcated related biological processes, molecular functions and cellular components, and KEGG pathway analysis enabled annotations of metabolic pathways and revealed interactions among the significantly enriched pathways. The results showed that the enriched GO term “Nervous system development” and enriched pathway “CCKR signaling map” might be important contributors to EV71-induced neuropathological mechanisms. In addition, we also screened 10 up- and down-regulated non-protein coding genes with significantly different expression in our transcriptome profiling, which suggested that these abnormally regulated non-protein-encoding genes might also play important roles in the pathogenesis of EV71 infection. Eventually, RT-qPCR technology was adopted to validate the transcriptome sequencing data and the experiment demonstrated that the RT-qPCR and transcriptome sequencing results were basically consistent. In summary, this is the first transcriptome analysis of SH-SY5Y cells in response to EV71 infection and provides valuable cues for further exploring the mechanism of nervous system changes caused by EV71 infection.

The direct replication of influenza virus is not the only cause of harm to human health; influenza infection leading to a hyper-inflammatory immune response can also result in serious conditions. So, the treatment strategy for influenza needs to keep balance between antivirus and anti-inflammation. Herein, we review the treatment strategies of anti-influenza drugs and traditional Chinese medicines.

Lead in paint is a major public health concern and has drawn much attention from international organizations such as the World Health Organization (WHO) and the United Nations Environmental Programme (UNEP). However, there is limited research on lead-based paint, especially its use in toys, which poses an explicit health risk to vulnerable children. The present study sampled 100 different toys purchased from the three largest online shopping platforms in China: JD, Taobao (TB) and Tmall (TM). These selected toys have been sold nearly 3 million times in total, suggesting that they may have reached the dwellings of millions of children. It was found that the toys sold by JD and TM, which are considered organized sellers, had average lead concentrations of 25 mg/kg and 32 mg/kg, respectively, much lower than that of toys sold by unorganized sellers on the TB platform, at 219 mg/kg. Approximately 12% of the toys purchased from TB contained paint with total lead concentrations exceeding China's regulatory standard for paints in toy manufacturing, and nearly 36% of the toys purchased from TB exceeded the equivalent US regulatory standard and EU standard. These results suggest that further action is needed to verify the health and safety standards of toys sold by such unorganized sellers. Moreover, this study found that China's regulatory standard (90 mg/kg) based on soluble lead may underestimate the risk posed by lead in paints, and it is suggested that future regulatory thresholds for lead levels in paints for toy manufacture be based on total rather than soluble lead concentrations. The present study also explored various influencing factors on lead concentration, and found that lead concentrations were related to toy price, age group, color, and sales volume.

The antidiabetic drug metformin exerts antineoplastic effects against breast cancer and other cancers. One mechanism by which metformin is believed to exert its anticancer effect involves activation of its intracellular target, adenosine monophosphate-activated protein kinase (AMPK), which is also implicated in the antidiabetic effect of metformin. It is proposed that in cancer cells, AMPK activation leads to inhibition of the mammalian target of rapamycin (mTOR) and the downstream pS6K that regulates cell proliferation. Due to its hydrophilic and cationic nature, metformin requires cation-selective transporters to enter cells and activate AMPK. This study demonstrates that expression levels of cation-selective transporters correlate with the antiproliferative and antitumor efficacy of metformin in breast cancer. Metformin uptake and antiproliferative activity were compared between a cation-selective transporter-deficient human breast cancer cell line, BT-20, and a BT-20 cell line that was engineered to overexpress organic cation transporter 3 (OCT3), a representative of cation-selective transporters and a predominant transporter in human breast tumors. Metformin uptake was minimal in BT-20 cells, but increased by >13-fold in OCT3-BT20 cells, and its antiproliferative potency was >4-fold in OCT3-BT20 versus BT-20 cells. This increase in antiproliferative activity was associated with greater AMPK phosphorylation and decreased pS6K phosphorylation in OCT3-BT20 cells. In vitro data were corroborated by in vivo observations of significantly greater antitumor efficacy of metformin in xenograft mice bearing OCT3-overexpressing tumors versus low transporter-expressing wildtype tumors. Collectively, these findings establish a clear relationship between cation-selective transporter expression, the AMPK-mTOR-pS6K signaling cascade, and the antiproliferative activity of metformin in breast cancer.

As one of the major pathogens in wound infections, Pseudomonas aeruginosa produces several virulence factors and forms biofilms; these processes are under the regulation of various quorum sensing (QS) systems. Therefore, QS has been regarded as a promising target to treat P. aeruginosa infections. In the present study, we evaluated the effect of the plant-derived QS inhibitor coumarin on P. aeruginosa biofilms and virulence. Coumarin inhibited QS in the P. aeruginosa QSIS2 biosensor strain, reduced protease and pyocyanin production, and inhibited biofilm formation in microtiter plates in different P. aeruginosa strains. The effects of coumarin in inhibiting biofilm formation in an in vitro wound model and reducing P. aeruginosa virulence in the Lucilia sericata infection model were strain-dependent. Transcriptome analysis revealed that several key genes involved in the las, rhl, Pseudomonas quinolone signal (PQS), and integrated QS (IQS) systems were downregulated in coumarin-treated biofilms of P. aeruginosa PAO1. Coumarin also changed the expression of genes related to type III secretion and cyclic diguanylate (c-di-GMP) metabolism. The cellular c-di-GMP level of P. aeruginosa PAO1 and recent clinical P. aeruginosa strains was significantly reduced by coumarin. These results provide new evidence for the possible application of coumarin as an anti-biofilm and anti-virulence agent against P. aeruginosa in wound infections.

Background A large body of evidence has shown that phthalate exposure can lower birth weight in animals and human beings. However, there are only limited data on whether phthalates could affect birth weight directly or indirectly through gestational age and pregnancy syndrome. Objectives To evaluate the effects of prenatal exposure to phthalates on birth weight in neonates and the mediation effects of gestational age and pregnancy syndrome on the association between phthalate exposure and birth weight. Methods In this study, 181 mother-newborn pairs were recruited from Wenzhou city. Maternal urine samples were collected during the third trimester and measured for phthalate metabolites by ESI-MS/MS. Structural equation models (SEMs) were used to evaluate effects of phthalate on birth weight controlling for maternal education, monthly income, nutritional supplements, infant gender, and maternal weight gain per week. The potential mediated effects of phthalate exposure through gestational age and pregnancy syndrome on birth weight were also calculated by structural equation modeling. Results After adjusting for potential confounders, urinary mono-phthalate levels (including MMP, MBP, MEHP, MEOHP, and MEHHP) were negatively associated with birth weight. A ten-fold increase in the concentration of MEOHP and MEHHP would be directly associated with lower birth weights (reduced to 124 g and 107 g, respectively). However, MBP had mediated effects on birth weight through gestational age, which was associated with an 85-g reduction in birth weight for every ten-fold increase in exposure. Both direct and mediated effects on birth weight were found in MMP and MEHP. The indirect effects of MMP and MEHP were mediated through gestational age and pregnancy syndrome. Thus, prenatal MMP and MEHP exposures were associated with decrease in birth weight. Conclusions A negative association exists between prenatal phthalate exposure and birth weight in Chinese neonates. In addition to direct pathway, phthalate exposures could affect birth weight through the mediated effects of gestational age and pregnancy syndrome.

Bacterial communication systems, such as quorum sensing (QS), have provided new insights of alternative approaches in antimicrobial treatment. We recently reported that one QS signal, named as autoinducer-2 (AI-2), can affect the behaviors of Pseudomonas aeruginosa PAO1 in a dose-dependent manner. In this study, we aimed to investigate the effects of AI-2 on P. aeruginosa PAO1 biofilm formation and virulence factors production in vitro, and in vivo using a pulmonary infection mouse model. Exogenous AI-2 resulted in increased biofilms architecture, the number of viable cells, and the yield of pyocyanin and elastase virulence factors in wild type P. aeruginosa PAO1. However, no such effect was observed in P. aeruginosa lasR rhlR mutant strain. In vivo, the use of AI-2 significantly increased the mortality, lung bacterial count and histological lung damage of mice with acute P. aeruginosa PAO1 infection. Our data suggest that AI-2 promotes the formation of P. aeruginosa PAO1 biofilms and the production of virulence factors by interfering with P. aeruginosa QS systems, resulting in decreased host survival. AI-2 may be a therapeutic target for the clinical treatment of a co-infection of P. aeruginosa and AI-2 producing bacteria.

Study objective Nonsteroidal anti-inflammatory drugs (NSAIDs) play a role in pain relief, especially in postoperative pain caused by inflammation. They have demonstrated significant opioid dose-sparing effects, which help in reducing postoperative effects and opioid side effects. The objective of this meta-analysis was to explore the role of NSAIDs in reducing postoperative pain at different time intervals and provide reference for medication after lumbar spine surgery by a meta-analysis of randomized controlled trials (RCT). Design A meta-analysis study of randomized controlled trials. Setting Postoperative recovery area. Patients Adult patients who have undergone lumbar spine surgery. Intervention Patients received NSAIDs for pain control after lumbar spine surgery. Measurements Standardized mean difference (SMD) and 95%CI were used to evaluate the visual analog scale of postoperative pain. Main results Four hundred and eight participants from eight studies were included in this study. The difference between the NSAIDs group and placebo is significant in 0–6, 12, and 24h groups (overall: SMD=−0.72, 95%CI −0.98 to −0.45; 0–6h: SMD=0.50, 95%CI −0.81 to −0.19; 12h: SMD=−1.07, 95%CI −1.45 to −0.70; 24h: SMD=−1.16, 95%CI −1.87 to −0.45). Heterogeneity and publication bias were observed in the 0–6 and 24h groups. Conclusion NSAIDs are effective in postoperative analgesia after lumbar spine surgery. The study type, NSAID dose, different surgery types, and analgesic type might influence the efficacy of NSAIDs.

Introduction The results presented by studies investigating the effect of remifentanil on both parturients and newborns during cesarean section differed significantly. Therefore, we performed a meta-analysis to estimate the effect of remifentanil on these patients. Evidence acquisition Potentially eligible studies published before 15 March 2016 were searched through four databases including PubMed, SCOPUS, ISI web of knowledge and EBSCO. Weighted mean difference (WMD) or odds ratios (ORs) and the corresponding 95% confidence interval (CI) were applied to estimate the strength of relationship. Evidence synthesis A total number of seven randomized-controlled trials were included in this meta-analysis. The results showed that Apgar values at 1 min and 5 min were significantly lower in the infants of remifentanil-treated mothers, with the WMD and corresponding 95% CI of -0.835 (-1.515, -0.154) and -0.296 (-0.570, -0.021), respectively. The pH value of umbilical artery was significantly higher in the remifentanil group (WMD: 0.014, 95% CI: 0.002, 0.025). The highest and lowest systolic blood pressures were significantly lower in remifentanil-treated mothers, with the WMD and corresponding 95% CI of -18.913 (-34.468, -3.359) and -12.982 (-21.479, -4.485), respectively. Conclusions Remifentanil shows potential value of maternal circulation response during general anesthesia, which reduces maternal blood pressure in response to intubation and surgery. However, whether it is beneficial for the neonate is still controversial. More randomized-controlled trials with larger sample size are required to assess the adverse effects of remifentanil.

Maternal exposure to particulate matter with aerodynamic diameter

Background Our previous research on the diversity of microbiota in the endotracheal tubes (ETTs) of neonates in the neonatal intensive care unit found that Pseudomonas aeruginosa (P. aeruginosa) and Streptococcus mitis (S. mitis) were the dominant bacteria on the ETT surface and the existence of S. mitis could promote biofilm formation and pathogenicity of P. aeruginosa. Toll-like receptor 4 (TLR4), which has been widely detected on the surface of airway epithelial cells, is the important component of the innate immune system. Therefore, we hypothesized that the co-existence of these two bacteria might impact the host immune system through TLR4 signaling. Results S. mitis rarely caused inflammation, whereas P. aeruginosa caused the most severe inflammation accompanied by increases in the number of inflammatory cells, interleukin (IL)-6 and tumor necrosis factor (TNF)-α expression, and total cell counts in BALF (p

Falls and their consequences are among the major health care problems affecting functional mobility and quality of life among older people. Even for people living independently in the community, falls are common occurrences (1,2). Although the causes of falls are varied and complex, a critical requirement needed for fall prevention is the ability to respond effectively to loss of balance. Protective stepping, an automatic reaction to instability, is a common strategy for maintaining balance in the everyday environment (3). Stepping is not a strategy of last resort but is often initiated well before the body approaches the limits of stability provided by the base of support (4) especially for older people (5,6). To effectively sustain standing balance, steps must be appropriately directed, timed, and executed to arrest the motion of the body center of mass by altering the base of support. Mounting evidence indicates that age-associated impairments in protective stepping contribute importantly to loss of balance and falls (3,6–8). An impaired ability to maintain lateral balance stability is especially relevant to falls among older people. For example, measures of mediolateral stability are associated with future falls (9) and past falls (10), and impaired control of lateral balance through stepping is particularly apparent for older adults (6,11,12). Furthermore, age-related falls often involve lateral body motion and associated hip fractures (13,14). Therefore, these observations underscore the specific challenges to maintaining balance and preventing falls that are linked with deficits in controlling lateral balance stability. Although there is accumulating support for the existence of a directional vulnerability to lateral instability and falls with aging, comprehensive and systematic investigations of this postulation are currently lacking. The aims of this study were (i) to determine the protective stepping response patterns evoked by different directions of externally applied waist-pull perturbations of standing balance and (ii) to identify the changes in stepping characteristics attributable to aging and in relation to the risk of falling. We hypothesized that the laterally oriented disturbances to balance would exacerbate the changes with age in protective stepping responses due to the increased demands for controlling lateral stability, and that these changes would be greater for those individuals who subsequently experienced falls.