Search

The aim of this study is to explore the perceptions of Chinese registered nurses on toxic leadership behaviours of nurse managers and to determine its type, cause and response measures.The nurse manager is the front-line leader of the nurses who provide services directly to patients. Previous evidence suggests that toxic leadership behaviours of nurse managers do exist and it is necessary to understand the specifics of it.We used phenomenological research methods to conduct semi-structured in-depth interviews among 12 nurses at a tertiary hospital in Wuhan over the period from January to March 2022. And the data were analysed using Colaizzi seven-step analysis method.Four themes were discovered: (a) nurses' perceptions of toxic leadership behaviours; (b) toxic leadership behaviours of nurse managers; (c) reasons for toxic leadership behaviours of nurse managers and (d) measures for toxic leadership behaviours of nurse managers.Chinese nurses are exposed to the toxic leadership of nurse managers for multiple reasons and respond differently.This study helps nursing managers identify which behaviours are harmful to the nurse that require special attention in developing strategies to buffer against nurse managers' toxic leadership.

Huijun Yue,1,* Yu Lin,1,* Yitao Wu,2,* Yongquan Wang,1 Yun Li,1 Xueqin Guo,1 Ying Huang,3 Weiping Wen,1 Gansen Zhao,2 Xiongwen Pang,2 Wenbin Lei1 1Otorhinolaryngology Hospital, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, People’s Republic of China; 2School of Computer Science, South China Normal University, Guangzhou, 510631, People’s Republic of China; 3Guangdong Province Traditional Chinese Medical Hospital, Guangzhou, 510000, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wenbin LeiOtorhinolaryngology Hospital, The First Affiliated Hospital, Sun Yat-sen University, 58 Zhongshan Road II, Guangzhou, Guangdong, 510080, People’s Republic of ChinaTel +86-13922113299Email leiwb@mail.sysu.edu.cnXiongwen PangSchool of Computer Science, South China Normal University, 55 West Zhongshan Avenue, Guangzhou, Guangdong, 510631, People’s Republic of ChinaTel +86-18620638848Email augepang@scnu.edu.cnPurpose: This study evaluated a novel approach for diagnosis and classification of obstructive sleep apnea (OSA), called Obstructive Sleep Apnea Smart System (OSASS), using residual networks and single-channel nasal pressure airflow signals.Methods: Data were collected from the sleep center of the First Affiliated Hospital, Sun Yat-sen University, and the Integrative Department of Guangdong Province Traditional Chinese Medical Hospital. We developed a new model called the multi-resolution residual network (Mr-ResNet) based on a residual network to detect nasal pressure airflow signals recorded by polysomnography (PSG) automatically. The performance of the model was assessed by its sensitivity, specificity, accuracy, and F1-score. We built OSASS based on Mr-ResNet to estimate the apnea‒hypopnea index (AHI) and to classify the severity of OSA, and compared the agreement between OSASS output and the registered polysomnographic technologist (RPSGT) score, assessed by two technologists.Results: In the primary test set, the sensitivity, specificity, accuracy, and F1-score of Mr-ResNet were 90.8%, 90.5%, 91.2%, and 90.5%, respectively. In the independent test set, the Spearman correlation for AHI between OSASS and the RPSGT score determined by two technologists was 0.94 (p < 0.001) and 0.96 (p < 0.001), respectively. Cohen’s Kappa scores for classification between OSASS and the two technologists’ scores were 0.81 and 0.84, respectively.Conclusion: Our results indicated that OSASS can automatically diagnose and classify OSA using signals from a single-channel nasal pressure airflow, which is consistent with polysomnographic technologists’ findings. Thus, OSASS holds promise for clinical application.Keywords: deep learning, nasal airflow, obstructive sleep apnea, residual network

The 5-year survival rate of laryngeal cancer continues to decline, and the laryngeal particularity of the anatomy adversely affects the patient's quality of life. Emerging evidence suggests that long noncoding RNAs (lncRNAs) are closely correlated to key steps in the malignant progression of cancer cells. In this study, we report the role of lncRNA SBF2-AS1/miR-302b-3p/TGFBR2 interactions in the metastasis of laryngeal squamous cell carcinoma (LSCC). We verified that SBF2-AS1 was significantly downregulated in LSCC tissues and cell lines using qRT-PCR analysis. Its low expression was correlated to lymph node metastasis and an advanced clinical stage. More importantly, LSCC patients with low expression of SBF2-AS1 tended to have a poor prognosis. Based on this, we performed gain-of-function and loss-of-function experiments in LSCC cell lines. The results confirmed that knocking down SBF2-AS1 can promote the metastasis of LSCC cells and enhance epithelial-mesenchymal transition phenotype, while the upregulation of SBF2-AS1 expression resulted in the opposite. Our in vivo model verified that SBF2-AS1 overexpression could inhibit LSCC cell metastasis. Subsequent mechanistic studies revealed that SBF2-AS1 acted as a competing endogenous RNA that upregulated the expression of TGFBR2 by endogenous sponging for miR-302b-3p in LSCC cell lines. Moreover, miR-302b-3p overexpression reversed the inhibitory effects on LSCC metastasis induced by upregulation of SBF2-AS1 expression, and inhibition of TGFBR2 expression reversed the effect of SBF2-AS1 on metastasis. Our study proposes SBF2-AS1 as a biomarker to predict the prognosis of LSCC patients and a novel potential therapeutic target.

Danjiangkou Reservoir is the water source of the mid-route of the South-to-North Water Transfer Project. The source, distribution and potential risk of antimony in its water and sediments are rarely reported. In this study, symmetrical investigation results demonstrated that the concentration of antimony in the Han sub-reservoir and water in front of the dam fluctuated at about 0.9 mg L−1, while it was relatively higher and increased with the distance from the dam in Dan sub-reservoir water, with an annual average of 0.93~3.15 mg L−1. In recent years, the concentration of antimony in the Danjiangkou Reservoir showed a downward trend, and the difference between the Han and Dan sub-reservoirs decreased significantly. The antimony in the sediments in the reservoir was primarily derived from the inflowing rivers, and it was higher in the Dan sub-reservoir than in the Han sub-reservoir. The concentration of antimony in the water in the reservoir was considerably higher than the background value in the watersheds, indicating that there is an external input with decreasing input intensity. The content of antimony in the sediments in the reservoir and its inflow rivers was substantially higher than the background value of watersheds, indicating that there is a certain degree of enrichment. In addition, the antimony mining industry in the water source area poses a risk to the water safety of the reservoir. Antimony is not a conventional pollutant. Consequently, the collection of antimony monitoring results is a challenging task. Additionally, this study fills the gap in regional antimony research. Furthermore, the ecological risk assessment of antimony in China is still in its infancy. Unquestionably, the study of the temporal and spatial distribution of antimony concentration will be beneficial for the protection of water sources in relevant regions.

To evaluate the performance of polymerase chain reaction (PCR)-free whole genome sequencing (WGS) for clinical diagnosis, and thereby revealing how experimental parameters affect variant detection.Five NA12878 samples were sequenced using MGISEQ-2000. NA12878 samples underwent WGS with differing deoxyribonucleic acid (DNA) input and library preparation protocol (PCR-based vs PCR-free protocols for library preparation). The depth of coverage and genotype quality of each sample were compared. The performance of each sample was measured for sensitivity, coverage of depth and breadth of coverage of disease-related genes, and copy number variants. We also developed a systematic WGS pipeline (PCR-free) for the analysis of 11 clinical cases.In general, NA12878-2 (PCR-free WGS) showed better depth of coverage and genotype quality distribution than NA12878-1 (PCR-based WGS). With a mean depth of ∼40×, the sensitivity of homozygous and heterozygous single nucleotide polymorphisms (SNPs) of NA12878-2 showed higher sensitivity (>99.77% and >99.82%) than NA12878-1, and positive predictive value exceeded 99.98% and 99.07%. The sensitivity and positive predictive value of homozygous and heterozygous indels for NA12878-2 (PCR-free WGS) showed great improvement than NA128878-1. The breadths of coverage for disease-related genes and copy number variants are slightly better for samples with PCR-free library preparation protocol than the sample with PCR-based library preparation protocol. DNA input also influences the performance of variant detection in samples with PCR-free WGS. All the 19 previously confirmed variants in 11 clinical cases were successfully detected by our WGS pipeline (PCR free).Different experimental parameters may affect variant detection for clinical WGS. Clinical scientists should know the range of sensitivity of variants for different methods of WGS, which would be useful when interpreting and delivering clinical reports.

Although it is well known that Bacillus thuringiensis Cry toxins kill insect pest by disrupting midgut cells of susceptible larvae through their pore formation activity, it is not clear what intracellular events are triggered after pore formation on the cell membrane of the target cells. Here we analyzed the role of Cry toxins on autophagy activation using several cell lines as models as well as in Helicoverpa armigera larvae. The selected insect cell lines (Hi5, Sl-HP and Sf9) were susceptible to activated Cry1Ca toxin, but only Sl-HP cells were also susceptible to activated Cry1Ac toxin. In contrast, the mammalian cell line 293 T was not susceptible to Cry1Ac or to Cry1Ca. Results show that Cry toxins induced autophagy only in the susceptible cell lines as shown by the analysis of the changes in the ratio of Atg8-PE to Atg8 and by formation of autophagosome dots containing Atg8-PE. The Cry1Ac enhanced autophagy in the midgut tissue of H. armigera larvae. Silencing expression of specific genes by RNAi assays confirmed that the autophagy induced by activated Cry toxins was dependent on AMPK and JNK pathways. Finally, inhibition of autophagy in the cell lines by specific inhibitors or RNAi assays resulted in delayed cell death triggered by Cry toxins, suggesting that the increased autophagy activity observed after toxin intoxication may contribute to cell death.

MotivationThe Coronavirus Disease 2019 (COVID-19) pandemic poses a huge threat to human public health. Viral sequence data plays an important role in the scientific prevention and control of epidemics. A comprehensive virus database will be vital useful for virus data retrieval and deep analysis. To promote sharing of virus data, several virus databases and related analyzing tools have been created.ResultsTo facilitate virus research and promote the global sharing of virus data, we present here VirusDIP, a one-stop service platform for archive, integration, access, analysis of virus data. It accepts the submission of viral sequence data from all over the world and currently integrates data resources from the National GeneBank Database (CNGBdb), Global initiative on sharing all influenza data (GISAID), and National Center for Biotechnology Information (NCBI). Moreover, based on the comprehensive data resources, BLAST sequence alignment tool and multi-party security computing tools are deployed for multi-sequence alignment, phylogenetic tree building and global trusted sharing. VirusDIP is gradually establishing cooperation with more databases, and paving the way for the analysis of virus origin and evolution. All public data in VirusDIP are freely available for all researchers worldwide.Availabilityhttps://db.cngb.org/virus/Contactweixiaofeng@cngb.org

The authors have withdrawn their manuscript whilst they perform additional experiments to test some of their conclusions further. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author.

With the application and development of high-throughput sequencing technology in life and health sciences, massive multi-omics data brings the problem of efficient management and utilization. Database development and biocuration are the prerequisites for the reuse of these big data. Here, relying on China National GeneBank (CNGB), we present CNGB Sequence Archive (CNSA) for archiving omics data, including raw sequencing data and its further analyzed results which are organized into six objects, namely Project, Sample, Experiment, Run, Assembly and Variation at present. Moreover, CNSA has created a correlation model of living samples, sample information and analytical data on some projects. Both living samples and analytical data are directly correlated with the sample information. From either one, information or data of the other two can be obtained, so that all data can be traced throughout the life cycle from the living sample to the sample information to the analytical data. Complying with the data standards commonly used in the life sciences, CNSA is committed to building a comprehensive and curated data repository for storing, managing and sharing of omics data. We will continue to improve the data standards and provide free access to open-data resources for worldwide scientific communities to support academic research and the bio-industry. Database URL: https://db.cngb.org/cnsa/.

Most of the variation in the human genome is a single nucleotide variation (SNV) based on a single base or small fragment insertions and deletions and genomic copy number variation (CNV). Both types of mutations are involved in many human diseases. Such diseases often have complex clinical symptoms and difficult clinical diagnosis, so an effective detection method is needed to help clinical diagnosis and prevent birth defects. With the development of sequencing technology, the method of chip capture combined with high-throughput sequencing has been extensively used because of its high throughput, high accuracy, high speed and low cost. This study designed a chip that captures the coding region of 3043 genes associated with 4013 monogenic diseases. In addition, 148 chromosomal abnormalities can be identified by setting targets in specific regions. Compared with the whole exon chip, the chip can detect 4013 monogenic diseases and 148 chromosomal abnormalities at a lower cost, including SNV, intra-gene CNV and genomic copy number variation. This study utilized a strategy of combining the BGISEQ500 sequencing platform with the chip to identify 102 disease-associated mutations in 63 patients, 69 of which were new mutations. The evaluation test results also show that this combination complies with the requirements of clinical testing and has good clinical application value.

Circulating tumor DNA (ctDNA) has been frequently investigated to monitor tumor dynamics and measure tumor burden. This non-invasive method concerning ctDNA has been recognized as a promising biomarker. Recently, next generation sequencing has been used in ctDNA detection by researchers. However, those reports have been limited by modest sensitivity, and only a minority of patients with cancer were applicable. Additionally, a limited number of cases of liver cancer have been analyzed. A more precise method is required to be established to evaluate ctDNA noninvasively. In the present study, a novel method to design a liver cancer-associated chip region (spanning 211 kb, containing 159 genes) was performed with high specificity using International Cancer Genome Consortium datasets. Following evaluation with datasets from The Cancer Genome Atlas and data from 3 patients with liver cancer, the selected regions were demonstrated to be beneficial to locate specific somatic mutations associated with liver cancer therapy and to monitor cancer dynamics in the plasma samples of the patients. In addition to establishing performance benchmarks supporting direct clinical use, the chip designed and the high-resolution sequencing analyses pipeline would allow the development a set of patient specific markers that could monitor the process of cancer with high accuracy and low cost. Furthermore, the present study is essential to understanding the dynamics and providing insight into the basic mechanisms of liver cancer.

Insecticidal proteins from Bacillus thuringiensis (Bt) are widely used to control insect pests, but their efficacy is reduced when pests evolve resistance. We report on a novel allele (r16) of the cadherin gene (PgCad1) in pink bollworm (Pectinophora gossypiella) associated with resistance to Bt toxin Cry1Ac, which is produced by transgenic cotton. The r16 allele isolated from a field population in China has 1545 base pairs of a degenerate transposon inserted in exon 20 of PgCad1, which generates a mis-spliced transcript containing a premature stop codon. A strain homozygous for r16 had 300-fold resistance to Cry1Ac, 2.6-fold cross-resistance to Cry2Ab, and completed its life cycle on transgenic Bt cotton producing Cry1Ac. Inheritance of Cry1Ac resistance was recessive and tightly linked with r16. Compared with transfected insect cells expressing wild-type PgCad1, cells expressing r16 were less susceptible to Cry1Ac. Recombinant cadherin protein was transported to the cell membrane in cells transfected with the wild-type PgCad1 allele, but not in cells transfected with r16. Cadherin occurred on brush border membrane vesicles (BBMVs) in the midgut of susceptible larvae, but not resistant larvae. These results imply that the r16 allele mediates Cry1Ac resistance in pink bollworm by interfering with the localization of cadherin.

© 2019 American Society of Human Genetics The occurrence of non-epileptic hyperkinetic movements in the context of developmental epileptic encephalopathies is an increasingly recognized phenomenon. Identification of causative mutations provides an important insight into common pathogenic mechanisms that cause both seizures and abnormal motor control. We report bi-allelic loss-of-function CACNA1B variants in six children from three unrelated families whose affected members present with a complex and progressive neurological syndrome. All affected individuals presented with epileptic encephalopathy, severe neurodevelopmental delay (often with regression), and a hyperkinetic movement disorder. Additional neurological features included postnatal microcephaly and hypotonia. Five children died in childhood or adolescence (mean age of death: 9 years), mainly as a result of secondary respiratory complications. CACNA1B encodes the pore-forming subunit of the pre-synaptic neuronal voltage-gated calcium channel Cav2.2/N-type, crucial for SNARE-mediated neurotransmission, particularly in the early postnatal period. Bi-allelic loss-of-function variants in CACNA1B are predicted to cause disruption of Ca2+ influx, leading to impaired synaptic neurotransmission. The resultant effect on neuronal function is likely to be important in the development of involuntary movements and epilepsy. Overall, our findings provide further evidence for the key role of Cav2.2 in normal human neurodevelopment.

In this work, low-cost and non-toxic diatomite was used as the silicon source and aluminum source to synthesize Al doped MCM-41 (Al-MCM-41), and Cs+ adsorbents has been prepared by immobilizing the potassium metal ferrocyanide (KMeFC, Me = Cu2+, Ni2+, Zn2+, Co2+, and Fe3+) on Al-MCM-41. The adsorbents have high surface area from 471.7 to 685.8 m2 g−1, and their Cs+ adsorption performance has been detailed investigated. The max Cs+ adsorption capacity is 1.1602 mmol g−1, 1.092 mmol g−1, 0.8174 mmol g−1, 0.6973 mmol g−1, and 0.4541 mmol g−1 for KZnFC/Al-MCM-41, KNiFC/Al-MCM-41, KCoFC/Al-MCM-41, KCuFC/Al-MCM-41, and KFeFC/Al-MCM-41, respectively. The KZnFC/Al-MCM-41 presents the highest adsorption capacity, and it can reach adsorption equilibrium with 10 min. The pH experiment results show the KZnFC/Al-MCM-41 can keep a higher adsorption capacity in pH range from 2 to 11. What more, the KZnFC/Al-MCM-41 present good adsorption selectivity against to K+, Na+, Ca2+, and Mg2+. The adsorption mechanism was also discussed, and the adsorption results show the adsorption process of Cs+ by KMeFC/Al-MCM-41 conforms to pseudo-second-order kinetics and Langmuir isotherm adsorption model, and the thermodynamic experiments confirmed the adsorption process of Cs+ by KMeFC/Al-MCM-41 are endothermic and spontaneous. The FT-IR, XRD, and XPS of KZnFC/Al-MCM-41 before and after adsorption of Cs+ inferred that the adsorption process of the adsorbents mainly completed by the exchange between Cs+ and K+. This work provide a novel strategy to synthesis of KZnFC/Al-MCM-41 in low-cost and green route, and the adsorbent as-synthesized in this work is an efficient candidate for removing or extracting Cs+ from radioactive wastewater and mineral resources.

Magnetic solid phase extraction (MSPE) was combined with hollow fiber liquid liquid liquid microextraction (HF-LLLME) for the extraction of seleno-amino acids. Graphene oxide (GO) modified magnetic nanoparticles (MNPs) were prepared and loaded with Cu2+ for the adsorption of target seleno-amino acids based on the interaction between Cu2+ and seleno-amino acids. Aqueous ethylenediamine was employed as the desorption solvent, and the desorption solution was directly employed as the donor solution in the subsequent HF-LLLME procedure for further preconcentration of target seleno-amino acids. In HF-LLLME, 1-octanol was employed as the extraction solvent, ionic liquid trioctylmethylammonium chloride ([MTOA]+[Cl]−) was employed as the carrier for the extraction of target seleno-amino acids, and aqueous NaNO3 solution was employed as the acceptor solution. The target seleno-amino acids were extracted from the donor solution into the organic phase and then back into the acceptor solution under the driving force of ion-pair formation between target seleno-amino acids and [MTOA]+[Cl]− and the gradient counter ions between the donor phase and the acceptor phase. Various factors influencing the extraction of target seleno-amino acids by MSPE and HF-LLLME were investigated thoroughly. Based on it, a method of MSPE-HF-LLLME coupled with high performance liquid chromatography (HPLC)-inductively coupled plasma mass spectrometry (ICP-MS) was developed and the analytical performance was evaluated under optimized conditions. After preconcentration by MSPE-HF-LLLME, the enrichment factors (EFs) for target seleno-amino acids ranged from 152 to 278-fold, and the LODs for target seleno-amino acids were in the range of 0.0075–0.013 μg L−1. A certified reference material of SELM-1 was used to validate the accuracy of the proposed method, and the determined value of SeMet was in good agreement with the certified values. The proposed method was successfully applied to the speciation of seleno-amino acids in rice and Se-enriched yeast cell samples.

Cotton bollworm (Helicoverpa armigera) is the major insect herbivore of cotton plants. As its larvae feed and grow on cotton, H. armigera can likely tolerate gossypol, the main defense metabolite produced by cotton plants, through detoxification and sequestration mechanisms. Recent reports have shown that various P450 monooxygenases and UDP-glycosyltransferases in H. armigera are involved in gossypol detoxification, while the roles of ABC transporters, another gene family widely associated with metabolite detoxification, remain to be elucidated. Here, we show that ingestion of gossypol-infused artificial diet and cotton leaves significantly induced the expression of HaABCB6 in H. armigera larvae. Knockdown and knockout of HaABCB6 increased sensitivity of H. armigera larvae to gossypol. Moreover, HaABCB6-GFP fusion protein was localized on lysosomes in Hi5 cells and its overexpression significantly enhanced gossypol tolerance in vitro. These experimental results strongly support that HaABCB6 plays an important role in gossypol detoxification by H. armigera.

The genetic basis of congenital mental retardation includes chromosomal anomalies and single gene mutations. In addition to chromosome microarray analysis, next‑generation sequencing (NGS) and Sanger sequencing have additionally been applied to identify single gene mutations. However, no methods exist to identify the cause of an anomaly in one step. The present study applied an improved targeted NGS method to diagnose an 8‑year‑old Chinese Han female with mental retardation in one step. The microdeletion 17p11.2 was successfully detected by the improved targeted NGS and no single gene mutations were identified. The same microdeletion was verified using low coverage whole‑genome sequencing. Fertility guidance was also given to the patient's parents. In the present study, an improved targeted NGS method was applied to diagnose non‑syndromic mental retardation of unknown cause in one step. This improved method has the potential to be developed into a screening panel for the effective diagnosis of genetic abnormalities in non‑syndromic mental retardation and other congenital anomalies.

A novel method by combining hollow fiber-liquid–liquid–liquid microextraction (HF-LLLME) with HPLC-ICP-MS was developed for the speciation of inorganic mercury (Hg2+) and organomercury including methylmercury (MeHg+), ethylmercury (EtHg+) and phenylmercury (PhHg+), in the micro-ecosystem of East Lake (Wuhan, China). The factors affecting HF-LLLME were systematically investigated. Under the optimized conditions, the limits of detection for four mercury species were in the range of 2.9–5.6 ng L−1 with the relative standard deviations (c = 0.5 μg L−1, n = 7) ranging from 5.6 to 10.6%. The developed method was validated by analyzing a certified reference material of DORM-2, and the determined values were in good agreement with the certified values. The method has been applied for the speciation of mercury in five representative samples from the micro-ecosystem of East Lake including atmospheric particles, lake water, fish, plant and sediment.

Purpose To identify disease-causing mutations in a Chinese patient with retinitis pigmentosa (RP). Methods A detailed clinical examination was performed on the proband. Targeted next-generation sequencing (NGS) combined with bioinformatics analysis was performed on the proband to detect candidate disease-causing mutations. Sanger sequencing was performed on all subjects to confirm the candidate mutations and assess cosegregation within the family. Results Clinical examinations of the proband showed typical characteristics of RP. Three candidate heterozygous mutations in 3 genes associated with RP were detected in the proband by targeted NGS. The 3 mutations were confirmed by Sanger sequencing and the deletion (c.357_358delAA) in PRPF31 was shown to cosegregate with RP phenotype in 7 affected family members, but not in 3 unaffected family members. Conclusions The deletion (c.357_358delAA) in PRPF31 was the disease-causing mutation for the proband and his affected family members with RP. To our knowledge, this is the second report of the deletion and the first report of the other 2 mutations in the Chinese population. Targeted NGS combined with bioinformatics analysis proved to be an effective molecular diagnostic tool for RP.

Porous anodic alumina (PAA) with highly ordered arrays of nanopores was prepared on Al fabrics or foils by a two-step anodization process. Studies on structural and thermal properties of the prepared PAA membranes were carried out. Scanning electron microscopy and transmission electron microscopy were performed on the prepared PAA membranes at room temperature and 600 °C. Photoluminescence (PL) properties of PAA on Al foils under different annealing temperatures (100–600 °C) and PAA on Al fabrics before-after dyeing by Rhodamine B (RhB) have been investigated. For PAA on Al foils, with the increase of the annealing temperature, the PL intensity increases first, which reaches a maximum value at 500 °C, and then it decreases. For PAA on Al fabrics after dyeing by RhB, the white sample changed to pink and a new peak at 580 nm in the PL curve was found.

The lack of control over microvasculature formation remains a key roadblock to the therapeutic vascularization and regeneration of functional tissues. In the current study, the integration of plasmid DNA (pDNA) condensation and electrospraying technologies was proposed to promote the regeneration of mature blood vessels through injectable or infusible administration of microparticles. Calcium phosphate (CP) nanoparticles with encapsulated plasmids encoding vascular endothelial growth factors (pVEGF) and basic fibroblast growth factor (pbFGF) were synthesized using reverse microemulsions. Electrosprayed microparticles with the loading of CP-pDNA nanoparticles were evaluated on both endothelial cells and smooth muscle cells and after subcutaneous infusion into animals. CP-pDNA nanoparticles was obtained with an average size of around 110 nm and electrosprayed into microparticles, resulting in high loading efficiency and extended protection on pDNA from external DNase environment. The inoculation of poly(ethylene glycol) into microparticle matrices realized a gradual release for 4 weeks of CP-pDNA nanoparticles, leading to an incremental transfection efficiency and strong secretion of extracellular matrices. After subcutaneous infusion of microparticles with encapsulated both CP-pVEGF and CP-pbFGF nanoparticles, significantly higher densities of blood vessels were achieved than those containing individual nanoparticles, and induced a rapid generation of mature blood vessels with few cytotoxicity and inflammation reactions. Electrosprayed microparticle with CP-pDNA nanoparticles encapsulated promoted the formation of vascular networks, providing clinical relevance for therapeutic vascularization and regeneration of functional tissues after injection to ischemic sites or entrapment into tissue engineering scaffolds.

The promotion of blood vessel initiation and growth plays an important role in the realization of therapeutic vascularization and regeneration of functional tissues. Astragalus membranaceus and angelica sinensis are commonly used traditional Chinese medicines for enriching the blood. In the current study astragaloside IV (AT, the main active ingredient of astragalus) and ferulic acid (FA, the main ingredient of angelica) were loaded into electrospun fibrous scaffolds to provide abundant and sustained biological factors required to initiate vascularization and bring it to maturity. The cell viability after AT and FA treatment was dose-dependent with an optimal concentration of around 50 μg/mL, and the most significant synergistic effect was demonstrated for the combined treatment with AT and FA with the ratio of 7/3 on both primary endothelial and smooth muscle cells. The in vitro release study showed that the amount of AT and FA release could be regulated by their loading amount and ratios in electrospun fibers. The localized and sustained codelivery of AT and FA indicated significantly high cell viability and secretion of extracellular matrices for both endothelial and smooth muscle cells, and induced significantly high densities of vascular structures after subcutaneous implantation. The most significant angiogenesis promotion with few inflammatory reactions was demonstrated for electrospun fibers containing AT and FA with the ratio of 7/3. It was suggested that the integration of the synergistic effect of Chinese medicine into electrospun fibrous scaffolds should provide clinical relevance for therapeutic vascularization, full vascularization in engineered tissues, and regeneration of blood vessel substitutes.

A new method of phase transfer hollow fiber liquid phase microextraction (PT-HF-LPME) combined with electrothermal vaporization inductively coupled plasma mass spectrometry (ETV-ICP-MS) has been developed for the determination of trace Co, Pd, Cd and Bi in environmental and biological samples. In PT-HF-LPME, an intermediate solvent (1-butanol) was added into the sample solution to ensure the maximum contact area between the target metal ions and the chelating reagent (8-hydroxyquinoline, 8-HQ), which accelerated the formation of 8-HQ-metal complexes and their subsequent extraction by extraction solvent (toluene). The experimental parameters affecting the extraction efficiency of PT-HF-LPME for the target metals were studied by simplex optimization and orthogonal array design (OAD) experiments. Under the optimized conditions, the enrichment factors for Co, Pd, Cd and Bi were 110, 393, 121 and 111-fold, respectively, the limits of detection (LODs, 3σ) ranged from 3.7 to 8.3 ng L(-1). The relative standard deviations (RSDs, c=0.5 ng mL(-1), n=7) were 8.7, 6.2, 12.4 and 12.9% for Co, Pd, Cd and Bi, respectively. To validate the accuracy of the proposed method, two Certified Reference Materials of GSBZ50009-88 Environment Water and GBW09103 Human Urine were analyzed, and the results obtained for Cd were in good agreement with the certified values. Finally, the developed method was successfully applied to the analysis of Co, Pd, Cd and Bi in lake water and human urine samples.

The analysis of rich catalogues of genetic variation from population-based sequencing provides an opportunity to screen for functional effects. Here we report a rare variant in APOC3 (rs138326449-A, minor allele frequency ~0.25% (UK)) associated with plasma triglyceride (TG) levels (−1.43 s.d. (s.e.=0.27 per minor allele (P-value=8.0 × 10−8)) discovered in 3,202 individuals with low read-depth, whole-genome sequence. We replicate this in 12,831 participants from five additional samples of Northern and Southern European origin (−1.0 s.d. (s.e.=0.173), P-value=7.32 × 10−9). This is consistent with an effect between 0.5 and 1.5 mmol l−1 dependent on population. We show that a single predicted splice donor variant is responsible for association signals and is independent of known common variants. Analyses suggest an independent relationship between rs138326449 and high-density lipoprotein (HDL) levels. This represents one of the first examples of a rare, large effect variant identified from whole-genome sequencing at a population scale., Population-based genome sequencing provides an increasingly rich resource for the identification of low-frequency, large effect variants associated with clinically important phenotypes. Timpson et al. use UK10K data to identify a variant of the APOC3 gene strongly associated with plasma triglyceride levels.

A new method of solidified floating organic drop microextraction (SFODME) combined with electrothermal vaporization (ETV)-inductively coupled plasma mass spectrometry (ICP-MS) was developed for the determination of trace heavy metals in environmental water samples with sodium diethyldithiocarbamate (DDTC) as both chelating reagent in SFODME and chemical modifier in ETV. The factors affecting the microextraction efficiency were studied in detail and the optimal extraction conditions were established. Under the optimal conditions, the limits of detection (LODs) for SFODME-ETV-ICP-MS determination of Co, Pd, Cd, Hg, Pb and Bi were found to be 0.0060, 0.0091, 0.0020, 0.0041, 0.0170 and 0.0041 ng mL(-1), respectively, with the relative standard deviations (RSDs) of 2.8-10.0% (c=0.5 ng mL(-1), n=7). The developed method was successfully applied to the analysis of six target metals in Yangtze River and East Lake water samples with recoveries ranging from 77.7 to 119.1%. To validate the accuracy of the method, a certified reference material of Environmental Water (GSBZ50009-88) was analyzed and the determined values were in good agreement with the certified values.

Challenges remain in the generation of heterogeneous tissues and the repairing of interfacial tissue between soft and hard tissues. The development of tissue engineering scaffolds with gradients in composition, structure, mechanical and chemical properties is essential to modulate cellular behaviors in a graded way and potentially support the growth of functionally graded tissues. Integrated with the three-dimensional (3-D) nanofibrous skeletal structure of native extracellular matrix, electrospun fibers with gradients in amino groups were generated in the current study through an aminolysis process by using a microinfusion pump. Gelatin grafts were constructed to create fibrous scaffolds with gradients in hydroxyapatite (HA) contents, crystal size and mechanical properties through in situ mineralization. Plasmid DNA (pDNA) was included during the mineralization process, and gradations in pDNA loading contents were created on fibrous scaffolds on the basis of HA gradients. Obvious gradients in cell density, osteoblastic differentiation and collagen deposition were demonstrated along the long axis of fibrous mats after cell seeding. Gradients in the amount of pDNA released and the expression of target proteins were indicated on the fibrous mats, which offered a temporally and spatially controlled delivery of growth factors in scaffolds. The creation of gradient futures on 3-D fibrous scaffolds may provide physical, chemical and biological cues and result in efficient regeneration of tissues with spatial distributions of the cell proliferation, differentiation, and matrix secretion.

Background Targeted next-generation sequencing (NGS) is a cost-effective approach for rapid and accurate detection of genetic mutations in patients with suspected genetic disorders, which can facilitate effective diagnosis. Methodology/Principal Findings We designed a capture array to mainly capture all the coding sequence (CDS) of 2,181 genes associated with 561 Mendelian diseases and conducted NGS to detect mutations. The accuracy of NGS was 99.95%, which was obtained by comparing the genotypes of selected loci between our method and SNP Array in four samples from normal human adults. We also tested the stability of the method using a sample from normal human adults. The results showed that an average of 97.79% and 96.72% of single-nucleotide variants (SNVs) in the sample could be detected stably in a batch and different batches respectively. In addition, the method could detect various types of mutations. Some disease-causing mutations were detected in 69 clinical cases, including 62 SNVs, 14 insertions and deletions (Indels), 1 copy number variant (CNV), 1 microdeletion and 2 microduplications of chromosomes, of which 35 mutations were novel. Mutations were confirmed by Sanger sequencing or real-time polymerase chain reaction (PCR). Conclusions/Significance Results of the evaluation showed that targeted NGS enabled to detect disease-causing mutations with high accuracy, stability, speed and throughput. Thus, the technology can be used for the clinical diagnosis of 561 Mendelian diseases.

Cigarette smoking is one of the most important risk factors for kidney cancer, but the molecular mechanism is poorly understood. To examine the expression change of histone H3 on lysine 27 trimethylase (H3K27me3) demethylases ubiquitously transcribed TPR gene on the X chromosome (UTX) in kidney cancer cell line 786-O after nicotine treatment, quantitative real-time-polymerase chain reaction and western blotting analysis were carried out. These results showed that nicotine can increase UTX messenger RNA and protein levels and also decrease the content of H3K27me3. The decreased content of H3K27me3 may activate specific gene expression and lead to kidney cancer. Future investigation on nicotine induced UTX expression and its epigenetic effect would deepen our understanding on nicotine toxicity and carcinogenicity.

Vascularization is one of the capital challenges in the establishment of tissue engineering constructs and recovery of ischemic and wounded tissues. The aim of this study was to assess electrospun fibers with loadings of multiple pDNA to allow a localized delivery for an efficient regeneration of mature blood vessels. To induce sufficient protein expression, a reverse microemulsion process was adopted to load pDNA into calcium phosphate nanoparticles (CP-pDNA), which were electrospun into fibers to achieve a sustained release for 4 weeks. Compared with pDNA-infiltrated fibers, the localized and gradual release of pDNA facilitated cell proliferation, gene transfection, and extracellular matrix secretion and enhanced the generation of blood vessels after subcutaneous implantation. Compared with commonly used pDNA polyplexes with poly(ethyleneimine), CP-pDNA nanoparticles induced significantly lower cytotoxicity and less inflammation reaction after implantation into animals. Fibers with encapsulated nanoparticles containing plasmids encoding vascular endothelial growth factor (pVEGF) and basic fibroblast growth factors (pbFGF) led to significantly higher density of mature blood vessels than those containing individual plasmid. It is suggested that the integration of CP-pDNA nanoparticles with loadings of multiple plasmids into fibrous scaffolds should provide clinical relevance for therapeutic vascularization, getting fully vascularized in engineered tissues and regeneration of blood vessel substitutes.

A novel method of ionic liquid based carrier mediated hollow fiber liquid liquid liquid microextraction (IL-carrier mediated HF-LLLME) combined with high performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS) has been developed for the speciation of five phenylarsenic compounds and arsenate in chicken and feed samples. The target arsenic species were extracted from 6 mL aqueous samples (pH = 10.2) into an organic phase (20% (v/v) [MTOA]+[Cl]− in toluene) impregnated in the pores of a hollow fiber, then back extracted into a 10 μL acceptor phase of 0.3 mol L−1 NaBr in the lumen of the hollow fiber. The main driving force for the extraction was the gradient concentration of the counter ion from the donor phase to the acceptor phase. Factors affecting the extraction of the target arsenic species were investigated in detail. Under the optimal conditions, the limits of detection (LODs, 3σ) for five target phenylarsenic compounds and arsenate were in the range of 1.4–16.0 ng L−1 with the relative standard deviations (CAs(V),4-OH,3-NHPAA,PA,4-NPAA = 0.05 μg L−1, Cp-ASA = 0.10 μg L−1, n = 7) ranging from 3.6 to 10.0%, and the enrichment factors were varied from 86 to 372-fold. The proposed method of IL-carrier mediated HF-LLLME-HPLC-ICP-MS was successfully applied to the determination of five phenylarsenic compounds and arsenate in chicken and feed samples with the recoveries of 79.2–105.4% and 81.1–117.8% for the spiked chicken and spiked feed samples, respectively.

Imputing genotypes from reference panels created by whole-genome sequencing (WGS) provides a cost-effective strategy for augmenting the single-nucleotide polymorphism (SNP) content of genome-wide arrays. The UK10K Cohorts project has generated a data set of 3,781 whole genomes sequenced at low depth (average 7x), aiming to exhaustively characterize genetic variation down to 0.1% minor allele frequency in the British population. Here we demonstrate the value of this resource for improving imputation accuracy at rare and low-frequency variants in both a UK and an Italian population. We show that large increases in imputation accuracy can be achieved by re-phasing WGS reference panels after initial genotype calling. We also present a method for combining WGS panels to improve variant coverage and downstream imputation accuracy, which we illustrate by integrating 7,562 WGS haplotypes from the UK10K project with 2,184 haplotypes from the 1000 Genomes Project. Finally, we introduce a novel approximation that maintains speed without sacrificing imputation accuracy for rare variants., Imputation uses genotype information from SNP arrays to infer the genotypes of missing markers. Here, the authors show that an imputation reference panel derived from whole-genome sequencing of 3,781 samples from the UK10K project improves the imputation accuracy and coverage of low frequency variants compared to existing methods.