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B10 cells are the most frequently investigated subset of Breg cells, capable of suppressing immunity through the expression of the immunosuppressive cytokine IL-10. B10 cells are enriched in phenotypically diverse B-cell subsets. Recently, CD9 was identified as a marker of B10 cells in mice (human B10 cells have a separate set of markers that do not overlap with murine B10 cells). Together with a combination of other B10 markers, CD9 can be used to distinguish both mature and immature B10 cells from nonregulatory B cells and support selective purification of B10 cells. Here we provide five methods for the characterization and activity evaluation of CD9 + B cells. The first method is used for the preparation of leukocytes, the second and third are used for the characterization of CD9 + B cells, while the last two methods serve to evaluate CD9 + B-cell activities. Finally, we detail the purification of RNA from B10 cells and the performance of transcriptomic assays.

A number of novel furo[2,3-b]quinoline derivatives were designed and synthesized by introducing different substituted anilines and phenols to C4-position of furo[2,3-b]quinoline. All target compounds were evaluated in vitro against two human breast cancer cell lines (MCF-7 and MDA-MB-231) and one normal breast cell (MCF-10A) by MTT (3-[4,5-dimethylthylthiazol-2-yl]-2,5 diphenyltetrazolium broide, Thiazolyl blue) method. Most derivatives showed significant cytotoxic activity on the two breast cancer cells with IC 50 values in the range of (5.60-26.24 μM) and a certain selectivity, especially in the inhibition of MDA-MB-231. More notably, they were less toxic to normal breast cell (MCF-7-10A). Compound I 7 could be considered as an ideal selective candidate for further study. Mechanism studies showed that I 7 could inhibit the proliferation of cells by arresting MDA-MB-231 cell cycle at G2/M phase. Overall, as a novel furo[2,3-b]quinoline derivative, I 7 exhibited an excellent inhibitory effect in MDA-MB-231 cell and was worthy of in-depth study., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background: Natural products and their molecular frameworks have been explored as invaluable sources of inspiration for drug design by means of structural modification, computer-aided drug design, and so on. Scopoletin extracting from multiple herbs exhibits potential anti-cancer activity in vitro and in vivo without toxicity towards normal cells., Objective: The study aims to obtain new scopoletin derivatives with enhanced anti-cancer activity. We performed chemical structure modification and researched the mechanism of anti-tumor activity., Methods: In this study, we considered scopoletin as a lead compound, designed and synthesized a series of scopoletin derivatives via introducing different heterocyclic fragments, and their chemical structures were characterized by NMR spectra ( 1 H NMR and 13 C NMR) and HRMS(ESI). The antiproliferative activity of target compounds in four cancer cell lines (MDA-MB-231, MCF-7, HepG2, and A549) was determined by the MTT assay. Compound 11b was treated with Ac-cys under different reaction conditions to explore the thiol addition activity of it. The Annexin V/PI and JC-1 staining assay were performed to investigate the anti-tumor mechanism of 11b., Results: Novel compounds 8a-h and 11a-h derivatives of scopoletin were synthesized. Most of the target compounds exhibited enhanced antiproliferative activity against different cancer cells and reduced toxicity towards normal cells. In particular, 11b displayed the optimal antitumor ability against breast cancer MDA-MB- 231 cells with an IC 50 value of 4.46 μM. Compound 11b also cannot react with Ac-cys under the experimental condition. When treated with 11b for 24 h, the total apoptotic cells increased from 10.8% to 79.3%. Besides, 11b induced the depolarization of mitochondrial membrane potential., Conclusion: Compound 11b was more active than other derivatives, indicating that the introduction of thiophene fragment was beneficial for the enhancement of antitumor effect, and it was also not an irreversible inhibitor based on the result that the α, β-unsaturated ketones of 11b cannot undergo Michael addition reactions with Accys. Furthermore, studies on the pharmacological mechanism showed that 11b induced mitochondrial depolarization and apoptosis, which indicated that 11b killed cancer cells via a mitochondrial apoptotic pathway. Therefore, in-depth research and structure optimization of this compound is warranted., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Excessive nitrogen (N) application is widespread in Southern China. The effects of N fertilization on soil properties and crop physiology are poorly understood in tropical red loam soil. We conducted a field experiment to evaluate the effect of nitrogen fertilization rates on physiological attributes (chlorophyll, plant metabolic enzymes, soluble matters) on banana leaves, soil properties (soil enzymes, soil organic matter (SOM), soil available nutrients) as well as banana crop yield in a subtropical region of southern China. The N rates tested were 0 (N 0 ), 145 (N 145 ), 248 (N 248 ), 352 (N 352 ), 414 (N FT ), and 455 (N 455 ) g N per plant. The correlations among soil factors, leaf physiological factors and crop yield were evaluated. The results indiated that the high rates of N fertilization (N FT and N 455 ) significantly decreased soil available potassium (K) content, available phosphorus (P) content, glutamine synthetase (GS) activity, and soluble protein and sugar contents compared with lower N rates. The N 352 treatment had the highest crop yields compared with higher N rates treatments, followed by the N 455 treatment. However, there were no significant differences in crop yields among N fertilization treatments. Factor analysis showed that the N 352 treatment had the highest integrated score for soil and leaf physiological factors among all treatments. Moreover, the N 352 treatment was the most effective in improving carbon and nitrogen metabolism in banana. Crop yield was significantly and positively linearly correlated with the integrated score ( r = 0.823, p < 0.05). Path analysis revealed that invertase, SOM and sucrose synthase (SS) had a strong positive effect on banana yield. Canonical correspondence analysis (CCA) suggested that available K, invertase, acid phosphatase and available P were the most important factors impacting leaf physiological attributes. Cluster analysis demonstrated distinct differences in N application treatment related to variations in soil and leaf factors. This study suggested that excessive N fertilization had a negative effect on soil fertility, crop physiology and yield. The lower N rates were more effective in improving crop yield than higher rates of N fertilization. The N rate of 352 g N per plant (N 352 ) was recommended to reduce excess N input while maintaining the higher yield for local farmers' banana planting., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2020 Sun, Li, Li, Chang, Zhang, Zeng, Zeng and Peng.)

The physical properties of two-dimensional (2D) materials depend strongly on the number of layers. Hence, methods for controlling their thickness with atomic layer precision are highly desirable, yet still too rare, and demonstrated for only a limited number of 2D materials. Here, we present a simple and scalable method for the continuous layer-by-layer thinning that works for a large class of 2D materials, notably layered germanium pnictides and chalcogenides. It is based on a simple oxidation/etching process, which selectively occurs on the topmost layers. Through a combination of atomic force microscopy, X-ray photoelectron spectroscopy, Raman spectroscopy, and X-ray diffraction experiments we demonstrate the thinning method on germanium arsenide (GeAs), germanium sulfide (GeS), and germanium disulfide (GeS 2 ). We use first-principles simulation to provide insights into the oxidation mechanism. Our strategy, which could be applied to other classes of 2D materials upon proper choice of the oxidation/etching reagent, supports 2D material-based device applications, e.g. , in electronics or optoelectronics, where a precise control over the number of layers (hence over the material's physical properties) is needed. Finally, we also show that when used in combination with lithography, our method can be used to make precise patterns in the 2D materials.

Aim: This meta-analysis, only including randomized controlled trials (RCTs), was conducted to assess separately and compare the therapeutic efficacy of adipose-derived mesenchymal stem cells (ADMSCs) and bone marrow-derived mesenchymal stem cells (BMSCs) for knee osteoarthritis (OA) at the same follow-up time. Methods: Potential relevant researches were identified from PubMed, Web of Science, Embase, Cochrane Library and clinicaltrials.gov. The data, from clinical trials concentrating on knee OA treated with ADMSCs or BMSCs, were extracted and pooled for meta-analysis to compare the clinical outcomes of patients with knee OA in visual analog scale (VAS), Western Ontario McMaster Universities Osteoarthritis Index (WOMAC), Lysholm knee scale (Lysholm) and Tegner activity scale (Tegner). Results: Nine randomized controlled trials including a total of 377 patients met the inclusion criteria. This meta-analysis obtained the following results. First, the improvement of VAS scores was statistically significant after BMSCs treatment at 6-, 12- and 24-month follow-up compared with control groups (p < 0.01). In contrast, the improvement of WOMAC scores was of no statistical significance, but showed a positive trend with the prolongation of the follow-up time (6 months: mean difference [MD] = 6.51; 95% CI: -2.38 to 15.40; p = 0.15; 12 months: MD = -6.81; 95% CI: -13.94 to 0.33; p = 0.06). Lysholm scores presented a similar pattern (12 months: MD = 1.93; 95% CI: -11.52 to 15.38; p = 0.78; 24 months: MD = 8.94; 95% CI: 1.45 to 16.43; p = 0.02). Second, VAS and WOMAC scores of patients after ADMSCs treatment were significantly improved at any follow-up time (p ≤ 0.05). The improvement of Lysholm scores was of no statistical significance compared with control groups, although treatment outcome at 12-month follow-up was better than that at 24-month follow-up, which was debatable because only data of one clinical trial were pooled in the analysis (12 months: MD = 7.50; 95% CI: -1.94 to 16.94; p = 0.12; 24 months: MD = 5.10; 95% CI: -3.02 to 13.22; p = 0.22). Finally, by comparing the statistical results of VAS and WOMAC scores, it could be concluded that the therapeutic effect of ADMSCs on knee OA was more effective than that of BMSCs. Conclusion: This meta-analysis showed that regeneration with BMSCs or ADMSCs had a great application potential in the treatment of patients with knee OA, and ADMSCs tended to be superior to BMSCs according to the limited clinical evidences available.

We report the fabrication and electrical characterization of germanium arsenide (GeAs) field-effect transistors with ultrathin channels. The electrical transport is investigated in the 20-280 K temperature range, revealing that the p-type electrical conductivity and the field-effect mobility are growing functions of temperature. An unexpected peak is observed in the temperature dependence of the carrier density per area at ∼75 K. Such a feature is explained considering that the increased carrier concentration at higher temperatures and the vertical band bending combined with the gate field lead to the formation of a two-dimensional (2D) conducting channel, limited to few interfacial GeAs layers, which dominates the channel conductance. The conductivity follows the variable-range hopping model at low temperatures and becomes the band-type at higher temperatures when the 2D channel is formed. The formation of the 2D channel is validated through a numerical simulation that shows excellent agreement with the experimental data.

Objectives: Hepatitis C virus (HCV) infection is associated with abnormal immune responses. Since regulatory T (Tregs) and B (Bregs) cells modulate the progression of infectious diseases, this study aimed at examining how these cells are involved with the development of HCV infection., Methods: The frequencies of circulating Bregs and Tregs were characterized using flow cytometry. Both the association and dynamic changes of these cells with related clinical parameters were analyzed after Direct-Acting Antiviral (DAA) agent treatments. Additionally, both regulatory B and T and naïve B and T cells were sorted and stimulated with healthy or HCV sera in vitro., Results: Bregs frequency in HCV-infected patients increased significantly and were positively correlated with levels of sera HCV RNA load, Alanine aminotransferase (AST) and total bilirubin (TBILI). Additionally, the increased Bregs returned to normal levels after DAA treatment. However, Tregs increased markedly in patients with HCV-cirrhosis and were significantly associated with Aspartate aminotransferase to Platelet Ratio Index (APRI) and Fibrosis 4 (FIB-4) scores. Furthermore, HCV sera doesn't expand either Tregs or Bregs, however, it does induce the IL-10 expression in B cells although it fails to induce FOXP3 expression in CD4 + T cells., Conclusions: Increased Bregs not only may be associated with poor viral eradication and liver injury but also may provide a predictive marker of HCV disease therapeutic efficacy following DAA-treatment. HCV sera may selectively induce Bregs. Tregs probably do not control disease status in the early stages but may contribute to the progression of liver fibrosis in the late stages of HCV infection., Competing Interests: None., (AJTR Copyright © 2020.)

To develop novel and efficient heat shock protein 90-cell division cycle 37 (Hsp90-Cdc37) interaction disruptors, several lipophilic fragments were introduced into celastrol (CEL) to synthesize 48 new CEL derivatives. Among all the target compounds, 41 was screened with superior antiproliferative activity on related cancer cells (IC 50 : 0.41-0.94 μM) and 41 could decrease the level of the Hsp90-Cdc37 complex in A549 cells. The capability to disrupt the Hsp90-Cdc37 interaction was stronger than that of CEL. Furthermore, pull-down assay, UV assay, and molecular docking analysis all showed that 41 might disrupt the interaction of the Hsp90-Cdc37 complex by preferentially binding to Cdc37 in cancer cells. Further studies showed that 41 could significantly regulate the levels of Hsp90-Cdc37 clients, thereby inducing the apoptosis of cancer cells. Together, 41 is a novel Hsp90-Cdc37 disruptor by binding to Cdc37 (hydrogen bond and/or covalent bond). Our results may provide reference for the discovery of effective Hsp90-Cdc37 disruptors.

Magnetotactic bacteria (MTB) are a large, polyphyletic group of aquatic microorganisms capable of absorbing large amounts of iron and synthesizing intercellular nano-scaled nanoparticles termed magnetosomes. In our previous transcriptomic studies, we discovered that a novel gene ( MGMSRv2&#95;2046 , termed as mg2046 ) in Magnetospirillum gryphiswaldense strain MSR-1 was significantly up-regulated during the period of magnetosome synthesis. In the present study, we constructed a MSR-1 mutant strain with deletion of mg2046 (termed Δmg2046 ) in order to evaluate the role of this gene in cell physiological status and magnetosome formation process. In comparison with wild-type MSR-1, Δmg2046 showed similar cell growth, but much lower cell magnetic response, smaller number and size of magnetosomes, and reduced iron absorption ability. mg2046 deletion evidently disrupted iron uptake, and redox equilibrium, and strongly inhibited transcription of dissimilatory denitrification pathway genes. Our experimental findings, taken together with results of gene homology analysis, indicate that Mg2046 acts as a positive regulator in MSR-1 under microaerobic conditions, responding to hypoxia signals and participating in regulation of oxygen metabolism, in part as a co-regulator of dissimilatory denitrification pathway with oxygen sensor MgFnr (MGMSRv2&#95;2946, termed as Mg2946). Mg2046 is clearly involved in coupled regulation of cellular oxygen, iron and nitrogen metabolism under micro-aerobic or anaerobic conditions. Our findings help explain how MSR-1 cells initiate dissimilatory denitrification pathway and overcome energy deficiency under microaerobic conditions, and have broader implications regarding bacterial survival and energy metabolism strategies under hypoxia.

Ischemic stroke is a common life-threatening disease. Epidemiological studies have shown that chronic periodontitis is closely related to ischemic stroke. However, it remains unknown whether periodontitis plays a direct role in the injury of cerebral ischemia. To explore the role of chronic periodontitis in the development process of ischemic stroke, we combined two mouse models: experimental periodontitis induced by a periodontal injection of lipopolysaccharide and ischemic stroke induced by the photothrombotic method. Alveolar bone loss and inflammatory infiltration of the periodontal tissue were found in the mice with experimental periodontitis. Periodontitis significantly increased the infarction volume, and numbers of activated microglia and astrocytes. Furthermore, an increased expression of nod-like receptor protein 3 inflammasome and interleukin-1β was detected in the peri-infarct region. We drew a conclusion that chronic periodontitis exacerbated ischemic stroke by increasing the activation of microglia/astrocytes and the expression of nod-like receptor protein 3 inflammasome and interleukin-1β. This suggested that chronic periodontitis played a role in ischemic brain injury directly through exacerbating the inflammation of the damaged brain.

Head and neck cancer is the 6th most common malignancy worldwide and urgently requires novel therapy methods to change the situation of low 5-years survival rate and poor prognosis. Targeted therapy provides more precision, higher efficiency while lower adverse effects than traditional treatments like surgery, radiotherapy, and chemotherapy. Blockade of PD-1 pathway with antibodies against PD-1 or PD-L1 is such a typical targeted therapy which reconstitutes anti-tumor activity of T cell in treatments of cancers, especially those highly expressing PD-L1, including head and neck cancers. There are many clinical trials all over the world and FDA has approved anti-PD-1/PD-L1 drugs for head and neck cancers. However, with the time going, the dark side of this therapy has emerged, including some serious side effects and drug resistance. Novel materials like nanoparticles and combination therapy have been developed to improve the efficacy. At the same time, standards for evaluation of activity and safety are to be established for this new therapy. Here we provide a systematic review with comprehensive depth on the application of anti-PD1/PD-L1 antibodies in head and neck cancer treatment: mechanism, drugs, clinical studies, influencing factors, adverse effects and managements, and the potential future developments.

Blood is a biological fluid that contains multiple blood fraction and cellular components. High-quality blood specimens are essential prerequisites for various downstream applications such as molecular epidemiology studies, genomics, and proteomics studies. Currently, protocols and research publications concerning the collection, handling, preservation, and stability of blood or blood fractions are constantly emerging. Moreover, standardized guidelines are a requirement for biorepositories to tightly control preanalytical variables originating from these procedures and obtain high-quality blood specimen for downstream analyses. In this review article, we summarize the best practices and fit-for-purpose protocols regarding blood collection, processing, storage, and stability. In addition, we present some typical quality biomarkers, which could be used to evaluate the integrity of blood specimens.

Tetraspanins are transmembrane proteins that modulate multiple diverse biological processes, including signal transduction, cell-cell communication, immunoregulation, tumorigenesis, cell adhesion, migration, and growth and differentiation. Here, we provide a systematic review of the involvement of tetraspanins and their partners in the regulation and function of B cells, including mechanisms associated with antigen presentation, antibody production, cytokine secretion, co-stimulator expression, and immunosuppression. Finally, we direct our focus to the signaling mechanisms, evolutionary conservation aspects, expression, and potential therapeutic strategies that could be based on tetraspanins and their interacting partners.

Oxoisoaporphine alkaloids are a family of oxoisoquinoline-derived alkaloids that were first isolated from the rhizome of Menispermum dauricum DC. (Menispermaceae). It has been demonstrated that oxoisoaporphine alkaloids possess various biological properties, such as cholinesterase and β-amyloid inhibition, acting as a topoisomerase intercalator, monoamine oxidase A inhibition, and are expected to become anti-Alzheimer's disease, anticancer, and antidepressant drugs. This review provides an overview of natural sources, synthetic routes, bioactivities, structure-function relationship, and modification investigations into oxoisoaporphine alkaloids, with the aim of providing references to the structure-activity relationships for the design and development of oxoisoaporphine derivatives with higher efficacy and therapeutic potential., (© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Protein-mediated endocytosis of membrane is a key event in biological system. The mechanism, however, is still not clear. Using a de novo designed bola-type peptide KKKLLLLLLLLKKK (K 3 L 8 K 3 ) as a protein mimic, we studied how it induced giant unilamellar vesicle (GUV) to form inward buds or endocytosis at varying conditions. Results show that the inward budding is initiated as the charged lipids are neutralized by K 3 L 8 K 3 , which results in a negative spontaneous curvature. If the charged lipids have unsaturated tails, the buddings are slim fibrils, which can further wrap into a spherical structure. In the case of saturated charged lipids, the buddings are rigid tubules, stable in the studied time period. The unsaturated lipid to saturated lipid ratio in the mother membrane is another key parameter governing the shape and dynamics of the buds. A complete endocytosis is observed when K 3 L 8 K 3 is attached with a hydrophobic moiety, suggesting that hydrophobic interaction helps the buds to detach from the mother membrane. The molecules in the surrounding medium, such as negatively charged oligonucleotides, are engulfed into the GUV via endocytosis pathway induced by K 3 L 8 K 3 . Our study provides a novel strategy for illustrating the endocytosis mechanism by using peptides of simple sequence.

Fusarium wilt of banana, which is caused by Fusarium oxysporum f. sp. cubense, is one of the most serious diseases of banana. F. oxysporum f. sp. cubense race 1 (Foc1) and race 4 (Foc4) are the most prevalent pathogens of banana cultivars in the world. To understand the differences in the infection processes between Foc1 and Foc4, green fluorescent protein-tagged strains of F. oxysporum f. sp. cubense tropical race 4 (FocTR4) and Foc1 were used to inoculate 'Brazil Cavendish' banana. At 2 days postinoculation (dpi), it was observed that the spores and hyphae of both Foc1 and Foc4 attached to the root hairs and root epidermis. At 3 dpi, the hyphae of both Foc1 and Foc4 were found in the vascular tissues of roots. However, Foc4 was observed in the parenchymal cells of banana root, whereas Foc1 was not found in parenchymal cells at 7 dpi. Furthermore, few Foc1 hyphae were observed in a few xylems whereas many more Foc4 hyphae were present in many xylems and phloems. Foc4 was observed in the vascular tissues of banana rhizomes, whereas no Foc1 was found in rhizomes 2 months after inoculation. The attachment process in F. oxysporum f. sp. cubense infection was further studied with scanning electron microscopy. Foc4 was observed to penetrate into banana roots from the intercellular space of the epidermis and wounds, whereas Foc1 mainly penetrated from the wounds but not from the intercellular space of the epidermis. Therefore, direct root penetration and rhizome vascular colonization by F. oxysporum f. sp. cubense are the key steps in the successful infection of Brazil Cavendish.

RNA is an essential biological material for research in genomics and translational medicine. As such, its storage for biobanking is an important field of study. Traditionally, long-term storage in the cold (generally freezers or liquid nitrogen) is used to maintain high-quality (in terms of quantity and integrity) RNA. Room temperature (RT) preservation provides an alternative to the cold, which is plagued by serious problems (mainly cost and safety), for RNA long-term storage. In this study, we evaluated the performance of several RT storage procedures, including the RNAshell ® from Imagene, where the RNA is dried and kept protected from the atmosphere, and the vacuum drying of RNA with additives such as the Imagene stabilization solution and a home-made trehalose solution. This evaluation was performed through accelerated (equivalent to 10 years for RNAshell) aging and real-time studies (4 years). To check RNA quality and integrity, we used RNA integrity number values and RNA-seq. Our study shows that isolation from atmosphere offers a superior protective effect for RNA storage compared with vacuum drying alone, and demonstrates that RNAshell permits satisfactory RNA quality for long-term RT storage. Thus, the RNA quality could meet the demand of downstream applications such as RNA-seq.

The stability of an electrodeposited nanocrystalline Ni-based alloy coating in a H₂S/CO₂ environment was investigated by electrochemical measurements, weight loss method, and surface characterization. The results showed that both the cathodic and anodic processes of the Ni-based alloy coating were simultaneously suppressed, displaying a dramatic decrease of the corrosion current density. The corrosion of the Ni-based alloy coating was controlled by H₂S corrosion and showed general corrosion morphology under the test temperatures. The corrosion products, mainly consisting of Ni₃S₂, NiS, or Ni₃S₄, had excellent stability in acid solution. The corrosion rate decreased with the rise of temperature, while the adhesive force of the corrosion scale increased. With the rise of temperature, the deposited morphology and composition of corrosion products changed, the NiS content in the corrosion scale increased, and the stability and adhesive strength of the corrosion scale improved. The corrosion scale of the Ni-based alloy coating was stable, compact, had strong adhesion, and caused low weight loss, so the corrosion rates calculated by the weight loss method cannot reveal the actual oxidation rate of the coating. As the corrosion time was prolonged, the Ni-based coating was thinned while the corrosion scale thickened. The corrosion scale was closely combined with the coating, but cannot fully prevent the corrosive reactants from reaching the substrate., Competing Interests: The authors declare no conflict of interest.

Stem cells are undifferentiated and pluripotent cells that can differentiate into specialized cells with a more specific function. Stem cell therapies become preferred methods for the treatment of multiple diseases. Oral and maxillofacial defect is one kind of the diseases that could be most possibly cured by stem cell therapies. Here we discussed oral diseases, oral adult stem cells, iPS cells, and the progresses/challenges/perspectives of application of stem cells for oral disease treatment.

Banana (Musa spp.) is an important staple and economic fruit crop, especially in Africa, Southeast Asia, and Latin America. The wilt disease caused by Fusarium oxysporum f. sp. cubense, especially F. oxysporum f. sp. cubense strain TR4, is disastrous for banana production. Banana plants infected by F. oxysporum f. sp. cubense TR4 gradually die from leaf blight or vascular rot. There is no efficient method to control this disease, and the underlying response of banana plants to F. oxysporum f. sp. cubense remains unknown. In this study, the responses of an economically important banana cultivar, the F. oxysporum f. sp. cubense-susceptible 'BX', and a wild banana relative, the F. oxysporum f. sp. cubense-resistant Musa yunnanensis ('YN'), to F. oxysporum f. sp. cubense infection were investigated using metabolomic, biochemical, and molecular biological methods. Numerous metabolomic compounds, including defense-responsive signaling molecules, phytohormones, phenolics, and antioxidants, were identified through metabolomic analysis. Changes in salicylic acid (SA), methyl-jasmonic acid, abscisic acid (ABA), cytokinin, 3-indoleacetic acid, gibberellic acid, and total phenolic levels were detected using liquid chromatography-mass spectrometry and the Folin-Ciocalteu method. The expression levels of genes involved in the biosynthesis of some defense-responsive compounds were studied through quantitative real-time polymerase chain reaction. The results revealed that the resistant YN had a larger change in SA content and a lower ABA level throughout the early infection period, compared with the levels in BX. The susceptible BX had a lower phenolic content. The resistant YN also expressed pathogenesis-related (PR) genes, especially PR1, PR4, PR5-1, and PDF2.2, at higher levels than the susceptible BX. These dynamic metabolic and gene-expression profiles from susceptible and resistant banana during the early stage of F. oxysporum f. sp. cubense infection increase our understanding of the complex interaction response between this crop and its pathogen.

Poly(methyl methacrylate) (PMMA) residue has long been a critical challenge for practical applications of the transferred chemical vapor deposited (CVD) graphene. Thermal annealing is empirically used for the removal of the PMMA residue; however experiments imply that there are still small amounts of residues left after thermal annealing which are hard to remove with conventional methods. In this paper, the thermal degradation of the PMMA residue upon annealing was studied by Raman spectroscopy. The study reveals that post-annealing residues are generated by the elimination of methoxycarbonyl side chains in PMMA and are believed to be absorbed on graphene via the π-π interaction between the conjugated unsaturated carbon segments and graphene. The post-annealing residues are difficult to remove by further annealing in a non-oxidative atmosphere due to their thermal and chemical stability. An electrolytic cleaning method was shown to be effective in removing these post-annealing residues while preserving the underlying graphene lattice based on Raman spectroscopy and atomic force microscopy studies. Additionally, a solution-gated field effect transistor was used to study the transport properties of the transferred CVD graphene before thermal annealing, after thermal annealing, and after electrolytic cleaning, respectively. The results show that the carrier mobility was significantly improved, and that the p-doping was reduced by removing PMMA residues and post-annealing residues. These studies provide a more in-depth understanding on the thermal annealing process for the removal of the PMMA residues from transferred CVD graphene and a new approach to remove the post-annealing residues, resulting in a residue-free graphene.

Lateral flow devices are versatile and serve a wide variety of purposes, including medical, agricultural, environmental, and military applications. Yet, the most promising opportunities of these devices for diagnosis might reside in point-of-care (POC) applications. Disposable paper-based lateral flow strips have been of particular interest, because they utilize low-cost materials and do not require expensive fabrication instruments. However, there are constraints on tuning flow rates and immunoassays functionalization in papers, as well as technical challenges in sensors' integration and concentration units for low-abundant molecular detection. In the present work, we demonstrated an integrated lateral flow device that applied the capillary forces with functionalized polymer-based microfluidics as a strategy to realize a portable, simplified, and self-powered lateral flow device (LFD). The polydimethylsiloxane (PDMS) surface was rendered hydrophilic via functionalization with different concentrations of Pluronic F127. Controlled flow is a key variable for immunoassay-based applications for providing enough time for protein binding to antibodies. The flow rate of the integrated LFD was regulated by the combination of multiple factors, including Pluronic F127 functionalized surface properties and surface treatments of microchannels, resistance of the integrated flow resistor, the dimensions of the microstructures and the spacing between them in the capillary pump, the contact angles, and viscosity of the fluids. Various plasma flow rates were regulated and achieved in the whole device. The LFD combined the ability to separate high quality plasma from human whole blood by using a highly asymmetric plasma separation membrane, and created controlled and steady fluid flow using capillary forces produced by the interfacial tensions. Biomarker immunoglobulin G (IgG) detection from plasma was demonstrated with a graphene nanoelectronic sensor integrated with the LFD. The developed LFD can be used as a flexible and versatile platform, and has the potential for detecting circulating biomarkers from whole blood. Sandwich-immunoassays can be performed directly on the LFD by patterning receptors for analytes on a desired substrate, and detections can be performed using a variety of sensing methods including nanoelectronic, colorimetric, or fluorescence sensors. The described bio-sensing technology presents an alternative for POC testing using small samples of human whole blood. It could benefit regions with limited access to healthcare, where delays in diagnosis can lead to quick deterioration of the quality of life and increase the morbidity and mortality., Competing Interests: Conflicts of Interest: The authors declare no conflict of interest.

A total of 16 targeted NO-releasing betulinic acid (BA) derivatives were designed and synthesized as potential anticancer agents. Most IC&lt;sub&gt;50&lt;/sub&gt; values were under 1.0 μM in vitro test against HepG2 and B16. The result suggested that derivatives of BA with α,β-unsaturated ketone skeleton possessed significant cytotoxic activities than the others, among which derivatives with three carbons in diol linker (15b and 15c) exhibited the highest anti-cancer activity. NO-releasing amount detection of partial target compounds suggested that NO-releasing amount of this series of BA derivatives positively correlates with their cytotoxic activities. The anti-angiogenic activity of partial target compounds on zebrafish embryos in our experiment did not show any effects on the SIVs, however, they exhibited different influence on ISVs, with only 15a and 15d better than the negative control.

Therapeutic effects of herbal medicines differ greatly due to the use of different anatomical parts or processing methods in traditional Chinese medicine, and Ephedra sinica (ES) is just a case in point. To better understand different traditional uses of the stems (known as Mahuang, MH) and roots (known as Mahuanggen, MHG) of ES, their therapeutic material basis should be investigated. In this study, ephedrine alkaloids were profiled simultaneously with primary metabolites using GC-MS based metabolomics. Ephedrine (E) has been reported to be the major bioactive constituent in MH for the treatment of asthma. The results showed that compared with MH, MHG contained much lower levels of five ephedrine alkaloids, which may well explain that MHG has not been used as an antiasthmatic. Additionally, these pharmacologically important ephedrine alkaloids exhibited strong positive correlation with five primary metabolites. In conclusion, this study facilitates better understanding of different traditional uses of MH and MHG., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Ling-Gui-Zhu-Gan decoction (LGZGD), a well-known traditional Chinese medicine (TCM) formula, has been extensively used for the treatment of cardiovascular disease in clinic. However, the chemical constituents in LGZGD had not been investigated so far. In this study, an ultra performance liquid chromatography-hybrid electrospray ionization linear ion trap-Orbitrap mass spectrometry (UPLC-LTQ-Oribitrap-MS/MS) method was established for rapid separation and structural identification of the constituents in LGZGD. Separation was performed on an ACQUITY(TM) UPLC BEH C18 column (50 × 2.1 mm, 1.7 μm) by gradient elution mode, using acetonitrile-water containing 0.1% formic acid as mobile phase at the flow rate of 0.2 mL/min. Accurate mass measurement for molecular ions and characteristic fragment ions could represent identification criteria for these compounds. As a result, 95 compounds including triterpene acids, triterpene saponins, flavonoids, coumarins, coumestans, benzofurans, phenylpropanoids and sesquiterpenoid lactones were detected, and 90 of them were tentatively identified. All compounds were further assigned in the individual raw material. In conclusion, the UPLC-LTQ-Orbitrap-MS/MS is a highly efficient technique to separate and identify constituents in complex matrices of TCMs. These results obtained in this research will provide a basis for quality control and further in vivo study of LGZGD., (© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Abstract Re-sequencing of target genes is a highly effective approach for identifying mutations in cancers. Mutations, including indels (insertions, deletions, and the combination of the two), play important roles in carcinogenesis. Combining genomic DNA capture using high-density oligonucleotide microarrays (NimbleGen, Inc.) with next-generation high-throughput sequencing, we identified approximately 1600 indels for colorectal cancers in the Chinese population. Among them, 5 indels were localized to exonic regions of genes, including the TFDP1 (transcription factor Dp-1) gene. TFDP1 is an important transcription factor that coordinates with E2F proteins, thereby promoting transcription of E2F target genes and regulating the cell cycle and differentiation. We report here the identification of a recurrent frame-shift indel mutation (named indel84) in the TFDP1 gene in colorectal cancers by next-generation sequencing. We found in a validation set that TFDP1 indel84 is present in 70% of colorectal cancer (CRC) tissues. Wild-type TFDP1 encodes a protein of 410 amino acids with a potential DNA binding site at its N-terminal followed by several functional protein domains. The TFDP1 indel cDNA would generate an alternative TFDP1 protein missing the first 120 amino acids and potentially affecting the DNA binding domain. We further demonstrated that the TFDP1 indel84 mutation generated a gain-of-function phenotype by increasing cell proliferation, migration, and invasion of CRC cells. Our study identified a key molecular event for CRC that might have great diagnostic and therapeutic potentials.

The phosphatase and transactivator EYA family proteins are overexpressed in many cancer cell lines and are abundantly distributed in undifferentiated cells during development. Loss-of-function studies have shown that EYA1 is required for cell proliferation and survival during mammalian organogenesis. However, how EYA1 is regulated during development is unknown. Here, we report that EYA1 is regulated throughout the cell cycle via ubiquitin-mediated proteolysis. The level of EYA1 protein fluctuates in the cell cycle, peaking during mitosis and dropping drastically as cells exit into G(1). We found that EYA1 is efficiently degraded during mitotic exit in a Cdh1-dependent manner and that these two proteins physically interact. Overexpression of Cdh1 reduces the protein levels of ectopically expressed or endogenous EYA1, whereas depletion of Cdh1 by RNA interference stabilizes the EYA1 protein. Together, our results indicate that anaphase-promoting complex/cyclosome (APC/C)-Cdh1 specifically targets EYA1 for degradation during M-to-G(1) transition, failure of which may compromise cell proliferation and survival.

Objective: To prepare and apply monoclonal antibodies (McAb) against recombinant human interferon alpha (rHu IFN-alpha)., Methods: Five cell lines (2E9, 4G1, 2A7, 2C9, 4G10) secreting McAbs against rHu IFN-alpha were established by hybridoma technique., Results: All the cell lines secreted monoclonal antibodies stably. Functions of secreting antibodies of the five cell lines lasted for 6 months in BALB/c mice and 8 months in cell culture. The specificity of antibody was constant. The Ig subclasses of the McAbs were IgG1. Anti-IFN McAb affinity purification column was prepared by coupling the anti IFN-alpha McAb to Sepharose 4B. The combining rate reached was higher than 95%., Conclusions: The highest purification efficiency was obtained by using 4G10 column.