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BackgroundLong-standing inflammatory bowel disease predisposes to the development of colorectal cancer (CRC). Interleukin (IL) -6, a pivotal link between chronic inflammation and tumor progression, has recently been recognized as a potential target. The effect of IL-6 on proliferation and metastasis of CRC by activating STAT3 pathway has been widely demonstrated in recent years, but few on mediating tumor immune evasion has been reported. MethodsIn this study, we analyzed IL-6 gene expression in human CRC tissues and carried out survival probability in CRC patients with TCGA Data portal and GEO Database. We studied CRC tumorigenesis in vivo by inoculating MC38 tumors and induced-CRC model via AOM/DSS （azoxymethan / dextran sulfate sodium）in (IL-6 deficient) IL-6-/- and (wild type) WT mice. We measured tumor growth and detected CD8+ T cells via immunofluorescence, we validated the function of IL-6 in tumorigenesis and tumor immune evasion in subcutaneous transplantation models. Using endoscopy monitoring and immunofluorescence, we validated the function of IL-6 in tumor immune evasion in AOM/DSS models. We conducted colitis model with DSS reveal the relationship of IL-6 and MHC-1 molecules by qRT-PCR and bioinformatics analysis. Organoid and CRC cells culture, western blot analysis, and qRT-PCR were used to investigate the expression and significance of MHC-1 molecules in vitro without IL-6 stimulation. MHC-1 molecules levels were also detected in tumors of IL-6-/- and WT mice from MC38-bearing tumor models and AOM/DSS models. Subcutaneous MC38 implantation models in C57BL/6 J mice were established to investigate anti-IL-6 therapy on CRC tumorigenesis. In vivo efficacy were conducted using antibodies blocking IL-6 and PD-1 in mice subcutaneous-bearing different types of tumors. Results Bio-information analysis and results of our center founded that IL-6 was remarkably overexpressed in CRC and its elevation was associated with poor prognosis. We observed that IL-6-/- mice were less susceptible to develop tumors, compared to (wild type) WT mice. Interestingly, infiltrated CD8+ T cells were found to exhibit high expression in tumor of IL-6-/- mice. High level of IL-6 was found in colitis model, with down-regulation of MHC-1molecules. Moreover, in vitro experiments, we found that IL-6 may act as a negative regulator in IFNγ-STAT1-MHC-1 signaling. In addition, vivo trials also confirmed that MHC-1 mRNA levels are negatively related to the existence of IL-6. Furthermore, blockade of IL-6 also activated CD8+ T-cell accumulation and led to the high PDL-1 expression in CRC, which can sensitize animals to anti-PD-1 therapy. Importantly, compared to treatment with anti-IL-6 alone and single treatment of anti-PD-1, combined IL-6 and PD-1 blockade greatly improved the therapeutic effect, not only in CRC but also melanoma and breast cancer.Conclusions Our study demonstrated that IL-6 act as a negative regulator in immunological surveillance to promote tumor immune evasion by inhibiting the response of cell to IFNγ. Blocking IL-6 enhanced tumor response to anti-PD-1 therapy via infiltration of CD8+ T cell and high expression of PDL-1. Based on the findings of our study, IL-6 is a novel target to improve the efficacy of immunotherapy.

Background Highly expressed STOML2 has been reported in a variety of cancers, yet few have detailed its function and regulatory mechanism. This research aims to reveal regulatory mechanism of STOML2 and to provide evidence for clinical therapeutics, via exploration of its role in colorectal cancer, and identification of its interacting protein. Methods Expression level of STOML2 in normal colon and CRC tissue from biobank in Nanfang Hospital was detected by pathologic methods. The malignant proliferation of CRC induced by STOML2 was validated via gain-of-function and loss-of-function experiments, with novel techniques applied, such as organoid culture, orthotopic model and endoscopy monitoring. Yeast two-hybrid assay screened interacting proteins of STOML2, followed by bioinformatics analysis to predict biological function and signaling pathway of candidate proteins. Target protein with most functional similarity to STOML2 was validated with co-immunoprecipitation, and immunofluorescence were conducted to co-localize STOML2 and PHB. Pathway regulated by STOML2 was detected with immunoblotting, and subsequent experimental therapy was conducted with RAF inhibitor Sorafenib. Results STOML2 was significantly overexpressed in colorectal cancer and its elevation was associated with unfavorable prognosis. Knockdown of STOML2 suppressed proliferation of colorectal cancer, thus attenuated subcutaneous and orthotopic tumor growth, while overexpressed STOML2 promoted proliferation in cell lines and organoids. A list of 13 interacting proteins was screened out by yeast two-hybrid assay. DTYMK and PHB were identified to be most similar to STOML2 according to bioinformatics in terms of biological process and signaling pathways; however, co-immunoprecipitation confirmed interaction between STOML2 and PHB, rather than DTYMK, despite its highest rank in previous analysis. Co-localization between STOML2 and PHB was confirmed in cell lines and tissue level. Furthermore, knockdown of STOML2 downregulated phosphorylation of RAF1, MEK1/2, and ERK1/2 on the MAPK signaling pathway, indicating common pathway activated by STOML2 and PHB in colorectal cancer proliferation. Conclusions This study demonstrated that in colorectal cancer, STOML2 expression is elevated and interacts with PHB through activating MAPK signaling pathway, to promote proliferation both in vitro and in vivo. In addition, combination of screening assay and bioinformatics marks great significance in methodology to explore regulatory mechanism of protein of interest.

For the different harvest targets, the requirement for the prolificacy trait of maize was also different, so prolificacy is of great significance for modern production. Although some QTLs and genes associated with prolificacy in teosinte have been reported, the genetic mechanism of prolificacy in maize has not been fully elucidated. In this study, two RIL populations and GWAS population were used to genetic research of prolificacy trait in maize, with multi-environment. Combine linkage analysis and Genome-wide association study has identified a total of 13 QTLs and 8 significant SNPs. There were two genes related to tissue differentiation in the stable QTL qP9-2, and two significant SNPs corresponding to three genes were in QTL qP5-1 and QTL qP7-1, respectively. Four candidate genes GRMZM2G317262, GRMZM2G317584, GRMZM5G882364 and GRMZM2G141679 were finally screened out by qRT-PCR analysis. Based on the function of candidate genes, ethylene signaling pathway plays an important role in the formation of prolificacy in maize. It has deepened our understanding of the formation mechanism of prolificacy and laid a foundation for breeding new varieties with various prolificacy in maize.

Scalable and cost-effective fabrication of three-dimensional (3D) boron carbon nitride (BCN) microspheres was first demonstrated by hydrothermal and annealing methods. In particular, the specific surface area of 3D-BCN-4 reached 1390.12 m2 g-1 and had a high hierarchical pore structure. An all-printed solid-state flexible microsupercapacitor (MSC) based on 3D-BCN-4 microspheres as an electrode material was fabricated for the first time by a screen printing method, which also provided efficacious properties. The single MSC areal capacitance reached 41.6 mF cm-2. Furthermore, the remarkable mechanical flexibility was also achieved for the device with evidence that no obvious capacitance loss occurred even upon bending to 180°, and the device had a 93.3% capacitance retention after 1000 cycles. In addition, the maximum energy density reached 0.00832 mW h cm-2, and the highest power density was 2 mW cm-2. These results show the synthesis of 3D-BCN by a facile and effective method with excellent electrochemical performance, which should provide a promising direction to wearable energy storage devices.

BackgroundOverexpression of STOML2 has been widely reported in a variety of cancer, yet few has detailed its function and regulatory mechanism. This study aims to reveal the clinicopathologic significance and oncologic function of STOML2 in colorectal cancer, explore its specific mechanism by means of yeast two-hybrid assay and bioinformatics.MethodsExpression level of STOML2 in normal colon and CRC tissue from biobank in Nanfang Hospital was detected by pathologic methods. The malignant proliferation of CRC induced by STOML2 was validated via gain-of-function and loss-of-function experiments, with novel techniques applied, such as organoid culture, orthotopic model and endoscopy monitoring. Yeast two-hybrid assay was conducted to screen interacting proteins of STOML2, followed by bioinformatics to predict biological process and signaling pathway of candidate proteins. Target protein with most functional similarity to STOML2 was validated with co-immunoprecipitation, and immunofluorescence were conducted to co-localize STOML2 and PHB. Pathway regulated by STOML2 was detected with immunoblotting, and subsequent experimental therapy was conducted with RAF inhibitor Sorafenib.ResultsSTOML2 was significantly overexpressed in colorectal cancer and its elevation was associated with unfavorable prognosis. Knockdown of STOML2 suppressed proliferation of colorectal cancer, thus attenuated subcutaneous and orthotopic tumor growth, while overexpressed STOML2 promoted proliferation in cell lines and organoids. A list of 13 interacting proteins was screened out by yeast two-hybrid assay. DTYMK and PHB were identified to be most similar to STOML2 according to bioinformatics in terms of biological process and signaling pathways; however, co-immunoprecipitation confirmed interaction between STOML2 and PHB, rather than DTYMK, despite its highest rank in previous analysis. Co-localization between STOML2 and PHB was confirmed in cell lines and tissue level. Furthermore, knockdown of STOML2 downregulated phosphorylation of RAF1, MEK1/2, ERK1/2 and ELK1 on the MAPK signaling pathway, indicating common pathway activated by STOML2 and PHB in colorectal cancer proliferation.ConclusionsThis study demonstrated that in colorectal cancer, STOML2 expression is elevated and interacts with PHB through activating MAPK signaling pathway, to promote proliferation both in vitro and in vivo. In addition, combination of screening assay and bioinformatics marks great significance in methodology to explore regulatory mechanism of protein of interest.

Purpose: To discuss the value and possible causes of γ-aminobutyric acid type A receptor-associated protein (GABARAP) expression and multiple clinicopathological factors in the prognosis of patients after radical pancreatic cancer treatment.Methods: The pancreatic tissues of 76 pancreatic cancer patients after R0 resection were screened according to the criteria, and the expression levels of GABARAP were determined by using immunohistochemistry (MaxVision) to label the pancreatic cancer tissues and normal pancreatic tissues at the peri-cancer level in these two types of specimens, and the relationship between GABARAP and other factors and the disease-free and overall survival of patients after radical pancreatic cancer treatment was evaluated by single factor survival analysis and Cox regression analysis.Results：The expression ratio of GABARAP in pancreatic cancer tissue was 55.26% (42/76), which was drastically higher than that in normal pancreatic tissue adjacent to cancer (21.42% (12/76). In 76 patients, the middle relapse-free survival time after radical operation for pancreatic cancer was 12.8 months, and the middle overall survival time was 17.8 months. Forty-one patients found relapse on CT, and the recurrence site: local recurrence accounted for 17.1% (7/41) Distant metastasis accounted for 78.0% (32/41), and local recurrence with distant metastasis accounted for 4.9% (2/41). Among the three recurrence methods, there was no drastical difference in survival time from relapse to death (P> 0.05). Cox regression model evaluation showed that GABARAP (P = 0.044) and postoperative adjuvant chemotherapy (P = 0.038) were the overall survival of patients after radical pancreatic cancer Period independent prognostic indicators and both are protective factors for the prognosis of pancreatic cancer patients. Further data analysis found that postoperative chemotherapy had no drastical effect on the relapse-free survival and overall survival of 42 GABARAP-positive patients with cancer tissue, while in 34 GABARAP-negative patients, postoperative chemotherapy drastically increased the total patient Survival period (P = 0.041) but had no drastical effect on the patient's relapse-free survival periodConclusion: The expression of GABARAP in pancreatic cancer tissues was drastically up-regulated, and patients with high expression of GABARAP in pancreatic cancer tissues had better prognosis, but had no drastical effect on the relapse-free survival of patients after radical operation of pancreatic cancer. The expression of GABARAP in pancreatic cancer tissues and Postoperative adjuvant chemotherapy is an independent indicator of patients' prognosis after radical pancreatic cancer resection. Both are protective factors. The high expression of GABARAP in pancreatic cancer may indicate that the adjuvant chemotherapy is low benefit.

In this paper, by precisely tuning the structure of composite, a 3D silkworm cocoon-like composite material based on reduced graphene oxide (RGO) and Z-CoO (with a ZIF-67 as precursor) are prepared through a in situ hydrothermal method. This Z-CoO/RGO nanocomposite exhibits a highly opened nanostructure and high conductive capability. The electrochemical performance reveals that an outstanding mass specific capacity (275 F g−1 at current density of 1 A g−1), which is much higher than the pure ZIF-67 with 2.7 F g−1, and excellent resistance characteristic are obtained due to the excellent synergistic effect between the MOFs and RGO. Moreover, a hybridized asymmetric supercapacitor with Z-CoO/RGO as anode and active carbon as cathode is constructed. The device shows ultra-low resistance (the equivalent-series resistance and the charge-transfer resistance are 1.13 Ω and 0.43 Ω, respectively) and stable cycle life about 1000 times is observed. This Z-CoO/RGO electrode also presents high current loading performance for possible high-power density type devices. Our work reveals that the straightforward hydrothermal treatment is a promising method for the preparation of dedicated MOFs complex nanostructure for high performance supercapacitor electrode applications.

Objective: To analyze the differential expression of related serum proteins was detected in patients with AR who were sensitized to dust mites pre- and 6-day post-RIT. Methods: Serum samples were collected from 15 AR, 15 6-day post-RIT and 10 healthy control patients. This analysis was performed using iTRAQ technique and was coupled with 2-dimensional gel electrophoresis and mass spectrometry. To determine the intimacy of the screened related differential proteins, a hierarchical analysis was performed with Cluster 3.0 software, and the results were visualized using Java TreeView software. Last, an ELISA was employed to validate the selected CRP, CTHRC1 and WDR89 protein expression data. Results: Compared with the healthy control group, the most upregulated protein in the AR group and the most downregulated proteins in the 6-day post-RIT group were CTHRC1 and WDR89. Compared with the healthy control group, the most downregulated protein in the AR group and the most upregulated protein of the 6-day post-RIT group was CRP. The serum protein levels of CRP, CTHRC1 and WDR89 proteins were determined by ELISA. It was found that the differential expression of CTHRC1 and WDR89 in the AR and 6-day post-RIT groups were statistically significant (P 0.05) was found in the differential expression of serum CRP in the AR and 6-day post-RIT groups. Conclusions: We found that CTHRC1 and WDR89 might be associated with the pathogenesis of AR, and they may play a regulatory role in the RIT.

In this work, we deposited Sb2S3 thin film with a benign molecular precursor ink based on alcohol solvent. Sb2S3 thin films with dense morphology, good crystallinity, and smooth surface were successfully prepared by optimizing the S/Sb ratio in the precursor. As-synthesized films have [hk1] preferred orientation, which is beneficial for the charge transport. The optical band gap of Sb2S3 thin film is inversely proportional to the S/Sb ratio in the precursor. This work provides a new idea and method for the application of Sb2S3 in photovoltaic.

The CC chemokine receptor 3 (CCR3) expressed by eosinophils, mast cells and Th2 cells is closely related to allergic diseases. The objective of this study was to explore whether silencing of CCR3 with short hairpin RNAs (shRNAs) delivered by a lentiviral vector could impact the function of mast cells in a murine model of allergic rhinitis (AR) in vivo. The murine model of allergic rhinitis (AR) inducing by ovalbumin (OVA) was constructed, and the BALB/c mice were divided into normal control group, AR group, controlshRNA treated group and lentiviral CCR3-shRNA treated group. The recombinant lentivirus vectors which express a short hairpin RNA (shRNA) targeting the CCR3 were dropped into the nasal cavity of OVA-sensitized mice before the challenges. Real-time fluorescence quantitative PCR and western blotting were performed to observe inhibitory effect of CCR3 gene. Nasal symptoms of mice and OVA-specific IgE in each group were assessed. Concentrations of histamine, tryptase and Prostaglandin D2 (PGD2) in bone marrow, peripheral blood and nasal mucosa were analyzed. Furthermore, histological analysis and electron microscopy analysis were applied to detect the histology changes of nasal mucosa and the infiltration of mast cells in nasal mucosa. The results showed that administration of CCR3shRNA could effectively inhibit the expression of the CCR3 gene in bone marrow, peripheral blood and nasal mucosa, which reduced the nasal symptoms, the level of OVA-specific IgE, the inflammatory cells and mast cells infiltration into nasal cavity, and relieved the histopathological changes of nasal mucosa. In addition, intervention of CCR3shRNA could reduce the levels of the histamine, tryptase and PGD2 in bone marrow, peripheral blood and nasal mucosa. These results suggest that inhibition of CCR3 gene expression by shRNAs lentiviral vectors can effectively attenuate migration, infiltration and degranulation of mast cells in local tissues and alleviate the inflammation of allergic rhinitis mice.

BackgroundThe mesocotyl connects the coleoptilar node and the basal part of the seminal root of maize (Zea mays) seedling. The mesocotyl pushes the shoot of the seedling out of the soil during seed germination; thus, its growth is highly related to deep-sowing tolerance. Although many studies on the maize mesocotyl have been carried out at physiological and molecular levels, the proteomic changes associated with cellular and physiological activities during mesocotyl growth are still unknown.ResultsIn the present study, the maize hybrid Zhengdan 958 was used to study mesocotyl growth and accompanying protein changes.The dark-grown etiolated mesocotyls exhibited a slow-fast-slow feature, with significant changes in the levels of indole-3-acetic acid (IAA) and cellulose and the activity of peroxidase (POD). In particular, POD activity increased with mesocotyl growth, showing higher activity at the mature (lower) end of the mesocotyl. For the proteomic analysis, soluble proteins were extracted from etiolated mesocotyls dark-grown for 48 h, 84 h, and 132 h, corresponding to the initial, rapid, and slow growth periods, respectively, and subjected to separation by two-dimensional gel electrophoresis (2-DE). As a result, 88 differentially abundant proteins (DAPs) were identified using MALDI-TOF-TOF analysis. At 48 h, most DAPs were stress proteins, heat shock proteins and storage proteins; at 84 h, oxidation/reduction proteins, carbohydrate biogenesis-related proteins and cytoskeleton-related proteins were highly accumulated; at 132 h, the most striking DAPs were those involved in the synthesis and modification of the cell wall and the biogenesis of carbohydrates. Gene ontology (GO) analysis showed that changes in the abundance and proportion of DAPs were consistent with cellular and physiological activities and biological processes during mesocotyl growth. The accumulation of nine DAPs of interest was verified by immunoblotting and RT-qPCR.ConclusionsThe present study revealed that the protein patterns in 2-D gels differed greatly with mesocotyl growth. At different growth periods, a specific set of DAPs participate in various biological processes and underlie the cellular and physiological activities of the mesocotyl. These results contributed to the understanding of mesocotyl growth and the cultivation of maize lines with deep-sowing tolerance.

[This corrects the article DOI: 10.1371/journal.pone.0202238.].

Additional file 3: Table S3. Primers used in RT-qPCR.

The cell wall is the most striking feature that distinguishes plant cells from animal cells. It plays an essential role in cell shape, stability, growth and protection. Despite being present in small amounts, cell wall proteins (CWPs) are crucial components of the cell wall. The cell wall proteome generally consists of sensu stricto CWPs, apoplast proteins and extracellular secreted proteins. Currently, there is a need for the bioinformatics analysis of a tremendous number of protein sequences that have been generated from genomic, transcriptomic and proteomics research. Compared with intracellular proteins, the location prediction of CWPs is challenging because many aspects of these proteins have not been experimentally characterized, and there are no CWP-trained, specific predictors available. By introducing the biological relevance (particularly molecular aspects) of the cell wall and CWPs, we critically evaluated the accuracy of 16 state-of-the-art predictors and classical predictors for the prediction of CWPs using an independent database of Arabidopsis and rice proteins. All experimentally verified CWPs and non-CWPs were retrieved from the UniProt Knowledgebase. Based on the evaluation, we currently recommend the predictors mGOASVM, HybridGO-Loc and FUEL-mLoc for CWPs. Furthermore, we outlined the public databases that can be used to cross-reference the subcellular location of CWPs. We illustrate a flowchart of the subcellular location prediction and a cross-reference of possible CWPs. Finally, we discuss challenges and perspectives in the bioinformatics analysis of CWPs. It is hoped that this article will provide practical guidance regarding CWPs for nonspecialists and provide insight for bioinformatics experts to develop computational tools for CWPs.

Protein extracts obtained from cells or tissues often require removal of interfering substances for the preparation of high-quality protein samples in proteomic analysis. A number of protein extraction methods have been applied to various biological samples. TCA/acetone precipitation and phenol extraction, a common method of protein extraction, is thought to minimize protein degradation and activity of proteases as well as reduce contaminants like salts and polyphenols. However, the TCA/acetone precipitation method relies on the complete pulverization and repeated rinsing of tissue powder to remove the interfering substances, which is laborious and time-consuming. In addition, by prolonged incubation in TCA/acetone, the precipitated proteins are more difficult to re-dissolve. We have described a modified method of TCA/acetone precipitation of plant proteins for proteomic analysis. Proteins of cells or tissues were extracted using SDS-containing buffer, precipitated with equal volume of 20% TCA/acetone, and washed with acetone. Compared to classical TCA/acetone precipitation and simple acetone precipitation, this protocol generates comparable yields, spot numbers, and proteome profiling, but takes less time (ca. 45 min), thus avoiding excess protein modification and degradation after extended-period incubation in TCA/acetone or acetone. The modified TCA/acetone precipitation method is simple, fast, and suitable for proteomic analysis of various plant tissues in proteomic analysis.