TGF-ß1 and IL-17A comediate the protumor phenotype of neutrophils to regulate the epithelial-mesenchymal transition in oral squamous cell carcinoma.

BACKGROUND: The role of neutrophils in cancer has been the subject of intense research in recent years. One major theme that has emerged is that not all neutrophils are equal in the field of cancer. However, it remains unclear what induces the protumorigenic or antitumorigenic phenotype predominate in tumor. Therefore, this study aimed to investigate what factors induce which of these two phenotypes of neutrophil predominate in OSCC and to explore the role of neutrophil polarization on tumor. METHODS: Immunofluorescence and immunohistochemistry staining were used to observe neutrophil infiltration and the expression of TGF-ß1 and IL-17A in OSCC tissues. Recombinant human TGF-ß1 and IL-17A were used to modulate neutrophil polarization. OSCC cell (SCC9 and SAS cell lines) migration, proliferation, invasion, stemness, and EMT were analyzed after treatment with conditioned medium from TGF-ß1/IL-17A-activated neutrophils. The levels of neutrophil-associated markers in OSCC tissues and peripheral blood were examined by immunofluorescence staining and quantitative PCR. RESULTS: Our data showed neutrophil infiltration and elevated expression of TGF-ß1 and IL-17A in OSCC tissues. The cooperative effect of TGF-ß1 and IL-17A promoted neutrophils to take on a protumor phenotype in vitro. TGF-ß1/IL-17A-activated neutrophils remarkably induced cell migration, proliferation, invasion, stemness, and EMT in OSCC cells. Additionally, OSCC patients showed increased expression of MMP9 and decreased expression of CCL3 in circulating neutrophils. CONCLUSION: TGF-ß1 and IL-17A cooperated to augment the protumor functions of neutrophils, thereby promoting the progression of OSCC cells. In addition, the combination of neutrophil-associated markers may serve as a predictive method to screen for patients with OSCC.

The large tegument protein pUL36 is essential for formation of the capsid vertex-specific component at the capsid-tegument interface of herpes simplex virus 1.

UNLABELLED: Herpesviruses have a characteristic particle structure comprising an icosahedral capsid, which contains the DNA genome and is, in turn, surrounded by a proteinaceous tegument layer and a lipid envelope. In herpes simplex virus, the interaction between the capsid and tegument is limited to the capsid vertices and involves two minor capsid proteins, pUL17 and pUL25, and the large inner tegument protein pUL36. pUL17 and pUL25 form a heterodimeric structure, the capsid vertex-specific component (CVSC), that lies on top of the peripentonal triplexes, while pUL36 has been reported to connect the CVSC to the penton. In this study, we used virus mutants with deletions in the genes for pUL36 and another inner tegument protein, pUL37, to analyze the contributions of these proteins to CVSC structure. Using electron cryomicroscopy and icosahedral reconstruction of mutants that express pUL17 and pUL25 but not pUL36, we showed that in contrast to accepted models, the CVSC is not formed from pUL17 and pUL25 on their own but requires a contribution from pUL36. In addition, the presence of full-length pUL36 results in weak density that extends the CVSC toward the penton, suggesting either that this extra density is formed directly by pUL36 or that pUL36 stabilizes other components of the vertex-tegument interface. IMPORTANCE: Herpesviruses have complex particles that are formed as a result of a carefully controlled sequence of assembly steps. The nature of the interaction between two of the major particle compartments, the icosahedral capsid and the amorphous tegument, has been extensively studied, but the identity of the interacting proteins and their roles in forming the connections are still unclear. In this study, we used electron microscopy and three-dimensional reconstruction to analyze virus particles formed by mutants that do not express particular interacting proteins. We show that the largest viral protein, pUL36, which occupies the layer of tegument closest to the capsid, is essential for formation of structurally normal connections to the capsid. This demonstrates the importance of pUL36 in the initial stages of tegument addition and provides new insights into the process of virus particle assembly.

[Progress in the application of laser ablation ICP-MS to surface microanalysis in material science].

In the present paper, apparatus and theory of surface analysis is introduced, and the progress in the application of laser ablation ICP-MS to microanalysis in ferrous, nonferrous and semiconductor field is reviewed in detail. Compared with traditional surface analytical tools, such as SEM/EDS (scanning electron microscopy/energy dispersive spectrum), EPMA (electron probe microanalysis analysis), AES (auger energy spectrum), etc. the advantage is little or no sample preparation, adjustable spatial resolution according to analytical demand, multi-element analysis and high sensitivity. It is now a powerful complementary method to traditional surface analytical tool. With the development of LA-ICP-MS technology maturing, more and more analytical workers will use this powerful tool in the future, and LA-ICP-MS will be a super star in elemental analysis field just like LIBS (Laser-induced breakdown spectroscopy).

Roles of Optineurin and Extracellular Vesicles in Glaucomatous Retinal Cell Loss.

OBJECTIVE: To explore the role of extracellular vesicles (EVs) in the pathogenesis of glaucoma caused by E50K mutation. METHODS: A photoreceptor cell line, RGC-5, was transfected with empty plasmids and plasmids expressing wild-type (WT) optineurin (OPTN) or E50K OPTN to investigate the effects of OPTN glaucoma as well as to identify the role of EVs in glaucoma pathology. The RGC-5 cells were also stimulated with glutamate, and their viability was evaluated using flow cytometry or CCK-8 assay. EVs were extracted, labeled with PKH-26, and added into the medium for normal RGC-5 culture, and the status of the cells was observed thereafter. RESULTS: WT OPTN overexpression, E50K OPTN, and glutamate stimulation induced apoptosis of RGC-5 cells. However, when glutamate stimulation was used as an add-on treatment, the degree of apoptosis in WT OPTN-overexpressing RGC-5 cells was significantly lower than that in E50K OPTN-expressing and normal RGC-5 cells. The viability of normal RGC-5 cells was reduced when co-cultured with WT OPTN-overexpressing RGC-5 or E50K OPTN-overexpressing RGC-5. EVs released by the latter two transfected lines similarly reduced normal RGC-5 survival. CONCLUSION: Our results indicate that WT OPTN overexpression may lead to photoreceptor apoptosis. However, overexpression also confers a degree of protection against high concentrations of extracellular glutamate. Additionally, EVs released by transfected RGC-5 cells may regulate the cell state. These findings may improve our understanding of the mechanisms of cell-cell interactions in pathological conditions, providing a basis for the use of EVs as novel targets for early diagnosis and treatment of glaucoma.

Pachymic Acid Ameliorates Pulmonary Hypertension by Regulating Nrf2-Keap1-ARE Pathway.

OBJECTIVE: Pulmonary hypertension (PH) is a severe pulmonary vascular disease that eventually leads to right ventricular failure and death. The purpose of this study was to investigate the mechanism by which pachymic acid (PA) pretreatment affects PH and pulmonary vascular remodeling in rats. METHODS: PH was induced via hypoxia exposure and administration of PA (5 mg/kg per day) in male Sprague-Dawley rats. Hemodynamic parameters were measured using a right ventricular floating catheter and pulmonary vascular morphometry was measured by hematoxylin-eosin (HE), α-SMA and Masson staining. MTT assays and EdU staining were used to detect cell proliferation, and apoptosis was analyzed by TUNEL staining. Western blotting and immunohistochemistry were used to detect the expression of proteins related to the Nrf2-Keap1-ARE pathway. RESULTS: PA significantly alleviated hypoxic PH and reversed right ventricular hypertrophy and pulmonary vascular remodeling. In addition, PA effectively inhibited proliferation and promoted apoptosis in hypoxia-induced pulmonary artery smooth muscle cells (PASMCs). Moreover, PA pretreatment inhibited the expression of peroxy-related factor (MDA) and promoted the expression of antioxidant-related factors (GSH-PX and SOD). Furthermore, hypoxia inhibited the Nrf2-Keap1-ARE signaling pathway, while PA effectively activated this pathway. Most importantly, addition of the Nrf2 inhibitor ML385 reversed the inhibitory effects of PA on ROS generation, proliferation, and apoptosis tolerance in hypoxia-induced PASMCs. CONCLUSION: Our study suggests that PA may reverse PH by regulating the Nrf2-Keap1-ARE signaling pathway.

A comprehensive comparison of circulating tumor cells and breast imaging modalities as screening tools for breast cancer in Chinese women.

Background: Circulating tumor cells (CTCs) have been recognized as a sensitive biomarker for breast cancer (BC). This study aimed to comprehensively compare CTC with imaging modalities, including ultrasonography, mammography, and contrast-enhanced magnetic resonance imaging (MRI) in screening for BC in Chinese women. Methods: Three hundred forty-three participants were enrolled in this study, including 102 treatment-naive BC patients, 177 with breast benign diseases (BBD) and 64 healthy female patients. All participants underwent CTC testing and at least one of the following examinations, ultrasonography, mammography, and MRI at the Second Affiliated Hospital of Zhejiang University between December 2017 and November 2020. CTCs were quantitatively assessed using cell counting (CTC detection rate/counts) and categorically examined using a cutoff value (CTC classification). The diagnostic power of CTC tests and imaging modalities, including accuracy and capability to predict clinicopathological characteristics of BC, were evaluated and compared. Results: CTC classification with a cutoff value of 2 showed a "good" diagnostic accuracy of 0.889 for early- to mid-stage BC comparable to breast imaging modalities using Breast Imaging-Reporting and Data System (BI-RADS). MRI demonstrated the highest sensitivity of 0.872 for BC, and CTC classification had the highest specificity of 0.938. A relatively low sensitivity was found for mammography in this cohort of patients. Successful detection of BC by CTC detection rate/counts, but not CTC classification, correlated with two important clinicopathological features, American Joint Committee on Cancer (AJCC) stage and tumor-node-metastasis (TNM) stage. The detection power of certain imaging modalities was also associated with AJCC stage (ultrasonography, p = 0.0438 and MRI, p = 0.0422) and lymph node metastasis (ultrasonography, 0.0157). There were clear correlations between CTC tests (counts or classification) and imaging BI-RADS scoring system in detecting positive BC cases (p < 0.05). Further correlation analysis suggested that CTC quantity, but not CTC classification, had the capability to predict clinicopathological traits of BC that were identified by ultrasonography. Conclusions: CTC tests have a diagnostic potency comparable to breast imaging modalities, and may be used as an alternative screening tool for BC.

The Melastoma dodecandrum genome and the evolution of Myrtales.

Melastomataceae has abundant morphological diversity with high economic and ornamental merit in Myrtales. The phylogenetic position of Myrtales is still contested. Here, we report the chromosome-level genome assembly of Melastoma dodecandrum in Melastomataceae. The assembled genome size is 299.81 Mb with a contig N50 value of 3.00 Mb. Genome evolution analysis indicated that M. dodecandrum, Eucalyptus grandis, and Punica granatum were clustered into a clade of Myrtales and formed a sister group with the ancestor of fabids and malvids. We found that M. dodecandrum experienced four whole-genome polyploidization events: the ancient event was shared with most eudicots, one event was shared with Myrtales, and the other two events were unique to M. dodecandrum. Moreover, we identified MADS-box genes and found that the AP1-like genes expanded, and AP3-like genes might have undergone subfunctionalization. The SUAR63-like genes and AG-like genes showed different expression patterns in stamens, which may be associated with heteranthery. In addition, we found that LAZY1-like genes were involved in the negative regulation of stem branching development, which may be related to its creeping features. Our study sheds new light on the evolution of Melastomataceae and Myrtales, which provides a comprehensive genetic resource for future research.

ICA512 signaling enhances pancreatic beta-cell proliferation by regulating cyclins D through STATs.

Changes in metabolic demands dynamically regulate the total mass of adult pancreatic beta-cells to adjust insulin secretion and preserve glucose homeostasis. Glucose itself is a major regulator of beta-cell proliferation by inducing insulin secretion and activating beta-cell insulin receptors. Here, we show that islet cell autoantigen 512 (ICA512)/IA-2, an intrinsic tyrosine phosphatase-like protein of the secretory granules, activates a complementary pathway for beta-cell proliferation. On granule exocytosis, the ICA512 cytoplasmic domain is cleaved and the resulting cytosolic fragment (ICA512-CCF) moves into the nucleus where it enhances the levels of phosphorylated STAT5 and STAT3, thereby inducing insulin gene transcription and granule biogenesis. We now show that knockdown of ICA512 decreases cyclin D1 levels and proliferation of insulinoma INS-1 cells, whereas beta-cell regeneration is reduced in partially pancreatectomized ICA512-/- mice. Conversely, overexpression of ICA512-CCF increases both cyclin D1 and D2 levels and INS-1 cell proliferation. Up-regulation of cyclin D1 and D2 by ICA512-CCF is affected by knockdown of STAT3 and STAT5, respectively, whereas it does not require insulin signaling. These results identify ICA512 as a regulator of cyclins D and beta-cell proliferation through STATs and may have implication for diabetes therapy.

Evaluation of circulating tumor cells as a prognostic biomarker for early recurrence in stage II-III breast cancer patients using CytoSorter® system: a retrospective study.

PURPOSE: Circulating tumor cells (CTCs) are known to be associated with late recurrence and poor prognosis in breast cancer (BC). Different CTC enrichment platforms have different CTC cut-off values for poor prognosis. This study aimed to evaluate whether preoperative CTCs could be a prognostic factor for early recurrence of disease in BC patients with resectable tumors, and to ascertain the CTC cut-off value for early recurrence with CytoSorter® CTC system. METHODS: Thirty-six stage II and III BC patients who had preoperative (pre-op) CTC detection and underwent a mastectomy or lumpectomy for curative intent between January and May 2018 were enrolled in this retrospective study. CTC detection was performed using CytoSorter® CTC system. Correlations of patients' demographics, clinicopathological characteristics, adjuvant therapies and CTCs with relapse and survival were evaluated. RESULTS: CTCs were detected in 32 out of 36 patients before surgery. Nine patients developed relapses during follow-up, and seven of them were distant recurrence. Univariate analysis showed that CTCs were correlated with two-year recurrence free survival (RFS) and distant RFS (D-RFS) (P = 0.013 and 0.029, respectively). Two-year RFS and D-RFS were 85.2% and 88.9%, respectively, for patients with <4 CTCs, while 44.4% and 55.6%, respectively, for patients with ≧4 CTCs. In multivariate analysis, only CTC was shown to be correlated with two-year RFS (HR: 0.219, 95% CI: [0.058-0.82], P = 0.024) and D-RFS (HR: 0.218, 95% CI [0.048-0.977], P = 0.047). CONCLUSION: BC patients with pre-op CTCs ≥4 per four mL of blood have significantly reduced two-year RFS and D-RFS. A pre-op CTC cut-off of four per four mL of blood was found for CytoSorter® to identify BC patients with a higher risk for early recurrence.

Comparison of circulating tumor cell (CTC) detection rates with epithelial cell adhesion molecule (EpCAM) and cell surface vimentin (CSV) antibodies in different solid tumors: a retrospective study.

PURPOSE: Status of epithelial-mesenchymal transition (EMT) varies from tumors to tumors. Epithelial cell adhesion molecule (EpCAM) and cell surface vimentin (CSV) are the most common used targets for isolating epithelial and mesenchymal CTCs, respectively. This study aimed to identify a suitable CTC capturing antibody for CTC enrichment in each solid tumor by comparing CTC detection rates with EpCAM and CSV antibodies in different solid tumors. METHODS: Treatment-naive patients with confirmed cancer diagnosis and healthy people who have performed CTC detection between April 2017 and May 2018 were included in this study. CTC detection was performed with CytoSorter® CTC system using either EpCAM or CSV antibody. In total, 853 CTC results from 690 cancer patients and 72 healthy people were collected for analysis. The performance of CTC capturing antibody was determined by the CTC detection rate. RESULTS: EpCAM has the highest CTC detection rate of 84.09% in CRC, followed by BCa (78.32%). CTC detection rates with EpCAM antibody are less than 40% in HCC (25%), PDAC (32.5%) and OC (33.33%). CSV has the highest CTC detection rate of 90% in sarcoma, followed by BC (85.71%), UC (84.62%), OC (83.33%) and BCa (81.82%). CTC detection rates with CSV antibody are over 60% in all 14 solid tumors. Except for CRC, CSV has better performances than EpCAM in most solid tumors regarding the CTC detection rates. CONCLUSION: EpCAM can be used as a target to isolate CTCs in CRC, LC, GC, BCa, EC, HNSCC, CC and PCa, especially in CRC, while CSV can be used in most solid tumors for isolating CTCs.

Enhancing the Screening Efficiency of Breast Cancer by Combining Conventional Medical Imaging Examinations With Circulating Tumor Cells.

PURPOSE: Ultrasound (US) and mammogram (MMG) are the two most common breast cancer (BC) screening tools. This study aimed to assess how the combination of circulating tumor cells (CTC) with US and MMG would improve the diagnostic performance. METHODS: CTC detection and imaging examinations, US and MMG, were performed in 238 treatment-naive BC patients, 217 patients with benign breast diseases (BBD), and 20 healthy females. Correlations of CTC, US and MMG with patients' clinicopathological characteristics were evaluated. Diagnostic performances of CTC, US and MMG were estimated by the receiver operating characteristic curves. RESULTS: CTC, US and MMG could all distinguish BC patients from the control (p < 0.0001). Area under curve (AUC) of CTC, US and MMG are 0.855, 0.861 and 0.759, respectively. While US has the highest sensitivity of 0.79, CTC and MMG have the same specificity of 0.92. Notably, CTC has the highest accuracy of 0.83. Combination with CTC increases the AUC of US and MMG to 0.922 and 0.899, respectively. Combining MMG with CTC or US increases the sensitivity of MMG to 0.87, however "CTC + MMG" has a higher specificity of 0.85. "CTC + US" performs the best in BC diagnosis, followed by "CTC + MMG" and then "US + MMG". CONCLUSION: CTC can be used as a diagnostic aid for BC screening. Combination with CTC increases the diagnostic potency of conventional BC screening imaging examinations, US and MMG, in BC diagnosis, especially for MMG.

Enhanced Performance of Fly Ash-Based Supports for Low-Cost Ceramic Membranes with the Addition of Bauxite.

Support is a necessary foundation for ceramic membranes to achieve high performance. Finding the optimum balance between high performance and low cost is still a significant challenge in the fabrication of ceramic supports. In this study, low-cost fly ash-based ceramic supports with enhanced performance were prepared by the addition of bauxite. The pore structure, mechanical strength, and shrinkage of fly ash/bauxite supports could be tuned by optimizing the bauxite content and sintering temperature. When the sintering temperature and bauxite content were controlled at 1300 °C and 40 wt%, respectively, the obtained membrane supports exhibited a high pure water permeance of approximately 5.36 m3·m-2·h-1·bar-1 and a high bending strength of approximately 69.6 MPa. At the same time, the optimized ceramic supports presented a typical mullite phase and excellent resistance to acid and alkali. This work provides a potential route for the preparation of ceramic membrane supports with characteristics of low cost and high performance.

Numerical Analysis of Oxygen-Related Defects in Amorphous In-W-O Nanosheet Thin-Film Transistor.

The integration of 4 nm thick amorphous indium tungsten oxide (a-IWO) and a hafnium oxide (HfO2) high-κ gate dielectric has been demonstrated previously as one of promising amorphous oxide semiconductor (AOS) thin-film transistors (TFTs). In this study, the more positive threshold voltage shift (∆VTH) and reduced ION were observed when increasing the oxygen ratio during a-IWO deposition. Through simple material measurements and Technology Computer Aided Design (TCAD) analysis, the distinct correlation between different chemical species and the corresponding bulk and interface density of states (DOS) parameters were systematically deduced, validating the proposed physical mechanisms with a quantum model for a-IWO nanosheet TFT. The effects of oxygen flow on oxygen interstitial (Oi) defects were numerically proved for modulating bulk dopant concentration Nd and interface density of Gaussian acceptor trap NGA at the front channel, significantly dominating the transfer characteristics of a-IWO TFT. Furthermore, based on the studies of density functional theory (DFT) for the correlation between formation energy Ef of Oi defect and Fermi level (EF) position, we propose a numerical methodology for monitoring the possible concentration distribution of Oi as a function of a bias condition for AOS TFTs.

Evaluation of Cell Surface Vimentin Positive Circulating Tumor Cells as a Diagnostic Biomarker for Lung Cancer.

BACKGROUND: Circulating tumor cells (CTCs) represent a collection of heterogeneous cells. Studies have shown epithelial CTCs and folate receptor (FR) positive CTCs could be used as diagnostic biomarkers for lung cancer (LC). This study aimed to determine whether cell surface vimentin (CSV) positive CTCs could be used as a biomarker for LC as well. METHODS: 78 treatment-naïve non-small-cell lung cancer (NSCLC) patients, 21 patients with benign lung diseases (BLD) and 9 healthy donors (HD) were enrolled in this study. CTC detection was performed using CytoSorter® mesenchymal CTC kit (CSV). The correlation between CSV positive CTCs (CSV-CTCs) and LC patients' clinicopathological characteristics would be evaluated, and diagnostic performances of CSV-CTCs and serum tumor markers for LC would be compared. RESULTS: CTC detection rates (average CTC count: range) in LC patients, patients with BLD and HD were 83.33% (2.47: 0-8), 47.62% (0.5: 0-3) and 0% (0: 0), respectively. CSV-CTCs could be used to differentiate LC patients from the patients with BLD and HD (P < 0.0001). CSV-CTCs were correlated with cancer stage, lymph node involvement and distant metastasis (P = 0.0062, 0.0014 and 0.0021, respectively). With a CTC cut-off value of 2, CSV-CTCs would have a sensitivity and specificity of 0.67 and 0.87, respectively, for diagnosing LC. CSV-CTC positive rates showed statistical differences among HD, BLD patients and LC patients at different cancer stages (P < 0.0001). Furthermore, CSV-CTC positive rates were positively correlated with tumor size, lymph node involvement and distant metastasis (P = 0.0163, 0.0196 and 0.03, respectively). CSV-CTCs had a better diagnostic performance than serum tumor makers, such as carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), cancer antigen 125 (CA125) and CA153. CONCLUSION: When CTC cut-off is set to 2 CTCs per 7.5 mL of blood, CSV-CTCs can be considered as an acceptable biomarker for diagnosing LC with a sensitivity and specificity of 0.67 and 0.87, respectively.

Screening for malachite green contamination on live fish skin with chewing gum based viscoelastic SERS sensor.

Malachite green (MG), a prohibited but still found antimicrobial in aquafarm and during live fish shipping, is a hot target in food safety screening. Herein, a novel chewing gum based flexible SERS (G-SERS) sensor was proposed for rapid sampling and detection of MG on live fish skin. The whole analysis takes <5 min, while the limit of detection for MG is 0.73 pg. Different from other reports, MG contaminated live fish was monitored daily with the G-SERS sensor, during which the fish was firstly raised in 0.5 ppm MG solution for one day, followed by freshwater for a week. It was found that the SERS signal of residue MG on fish skin could still be seen even on the sixth day, roughly the sale cycle of live fish in a marketplace. Furthermore, the method was also applied for MG screening on the skin of fish purchased from a supermarket and a local street marketplace. MG was found on some fishes from the latter but not from the former, which was cross-validated by LC-MS, suggesting MG risks still exist in smaller marketplaces. This work demonstrated the feasibility of using the flexible SERS sensor for onsite food safety screening.

Downregulation of FOXP3 in neutrophils by IL-8 promotes the progression of oral squamous cell carcinoma.

The aim of the present study was to investigate the effects of the transcription factor forkhead box P3 (FOXP3) in neutrophils on the progression of oral squamous cell carcinoma (OSCC). Cancer tissue samples and paracarcinoma tissues were collected from 23 patients with OSCC for the current study. In addition, SCC-9, a human tongue carcinoma cell line, was co-cultured with primary human neutrophils and treated with recombinant interleukin 8 (IL-8). The effect of FOXP3 on the proliferation of SCC-9 cells was analyzed using a Cell Counting Kit 8 assay. FOXP3 expression in neutrophils was analyzed by quantitative PCR following IL-8 treatment. FOXP3 protein expression in neutrophils and the amount of IL-8 protein in the OSCC tumor microenvironment were determined by immunofluorescence analysis. The present study demonstrated that IL-8 downregulated FOXP3 mRNA expression in neutrophils. Neutrophils and peptide P60, a specific inhibitor of FOXP3, increased proliferation of SCC-9 cells. In patients with OSCC, FOXP3 protein expression in neutrophils of the stage IV group was significantly lower compared with that of the stage II and stage III groups, while IL-8 protein expression was higher in cancer tissues compared with that in paracarcinoma tissues. In summary, IL-8 in the tumor microenvironment may recruit neutrophils, and downregulation of FOXP3 in neutrophils by IL-8 may promote the progression of OSCC.

NS5ATP9, a gene up-regulated by HCV NS5A protein.

Non-structural protein 5A (NS5A) appears to interact with a variety of cellular proteins and play an important role in mediating cell growth, cellular signaling pathways and pathogenesis of hepatitis C virus (HCV). NS5ATP9 was identified as a NS5A trans-activated protein in suppression subtractive hybridization (SSH), and the regulation was confirmed by luciferase reporter assay and quantitative real time PCR (qRT-PCR). A minimal promoter region contained within the 211bp (nucleotides -161 to +50bp) immediately upstream of the transcription initiation site. NS5ATP9 is a NS5A up-regulation gene which may play a role in the pathogenesis of HCV-associated hepatocellular carcinoma.

Expression of matrix metalloproteinase-8 and matrix metalloproteinase-13 in mast cells of human periapical lesions.

OBJECTIVE: This study aimed to detect the expression of matrix metalloproteinase-8 (MMP-8) and MMP-13 in mast cells (MCs) of human periapical lesions and to discuss the pathogenic role of MCs in periapical lesions. METHODS: Ninety samples were divided into three groups: (1) periapical granuloma group (n=30); (2) periapical cyst group (n=30); (3) normal periodontal membrane group (n=30). The samples were fixed in 10% neutral formalin for over 48 h and made into serial sections. After H&E staining, histological changes were observed under the optical microscope. Moreover, double immunofluorescence (DIF) staining was performed to detect expression of MMP-8 and MMP-13 in MCs of periapical lesions under the fluorescence microscope. RESULTS: Compared with the normal control group, the number of MMP-8 and MMP-13 double positive MCs in the periapical lesions increased significantly (P<0.01). There was no significant difference in the density of MMP-8 and MMP-13 double positive MCs in the periapical cyst group and periapical granuloma group (P>0.05). CONCLUSION: The number of MCs increased significantly in periapical lesions and there was a considerable increase in the density of MMP-8 and MMP-13 double positive MCs. These results indicate that MCs positive for MMP-8 and MMP-13 might contribute to the pathogenesis of chronic apical periodontitis.

[Screening, identification and biocontrol effect of antagonistic actinomycetes against the pathogen of Cytospora sp. for apple tree]. / è‹¹æžœæ ‘è çƒ‚ç— èŒæ‹®æŠ—æ”¾çº¿èŒçš„ç­›é€‰ã€é‰´å®šåŠé˜²æ•ˆ.

The pathogen of Cytospora sp. was considered as the target fungus to evaluate the bio-control potential of antagonistic actinomycetes against Cytospora sp. in the present study, which was isolated from the rhizosphere soil of apple tree. Moreover, the antagonistic effect of actinomycetes against Cytospora sp. growth was determined by the method of confrontation and growth rate, and the screened antagonistic strain was identified by the method of morphology and molecular biology. Also, the inhibitory activity of strain ZZ-9 against the conidium germination and mycelia growth of Cytospora sp. was determined in vivo, and its bio-control effect on Cytospora sp. growth was determined in detached apple twigs. The results showed that fifteen strains of actinomycetes had the inhibitory effect on Cytospora sp. growth, among all the isolated strains, which accounted for 18.8% and especially the inhibitory rate of eight strains was more than 50%. In addition, among all the screened strains, the strain ZZ-9 presented the highest inhibitory rate of 96.4%, which was significantly higher than those of the other isolated strains. The strain ZZ-9 was initially identified as Streptomyces rochei according to cultural characteristics, physiological and biochemical properties and 16S rDNA analysis. The nucleotide sequences in GenBank accession number was registered as KT986228. Different dilution of ZZ-9 fermentation had significant inhibitory effect on Cytospora sp. conidium germination and mycelia growth. The inhibitory rates of the 50 times fermentation on both conidium germination and mycelia growth were more than 80%. The inhibited mycelia's color was deepened, the mycelia branches were increased, and the ends of hyphae were swelled and deformed, even the protoplasm was concentrated and released. The bio-control effect of the ZZ-9 stock solution on Cytospora sp. growth was more than 75% in detached apple twigs. Thus, our results indicated that the strain ZZ-9 could be used for controlling apple tree Valsa canker in vivo and vitro.

Knockdown of FRAT1 inhibits hypoxia-induced epithelial-to-mesenchymal transition via suppression of the Wnt/ß-catenin pathway in hepatocellular carcinoma cells.

Hypoxia-induced epithelial-to-mesenchymal transition (EMT) in hepatocellular carcinoma (HCC) was investigated. Frequently rearranged in advanced T-cell lymphomas-1 (FRAT1) is a positive regulator of the Wnt/ß-catenin signaling pathway and is overexpressed in many human tumors. However, the expression and role of FRAT1 in HCC has not been elucidated. In this study, we investigated the effect of FRAT1 on EMT process in HCC cells induced by hypoxia. Our results showed that FRAT1 is highly expressed in HCC tissues and cell lines. Hypoxia significantly induced FRAT1 expression in HCC cells. FRAT1 knockdown inhibited hypoxia-induced cell migration/invasion, downregulation of epithelial markers and upregulation of mesenchymal markers. Moreover, FRAT1 knockdown suppressed the expression levels of ß-catenin, cyclin D1 and c-myc in HCC cells under the same hypoxic condition. Our results revealed that FRAT1 is a hypoxia factor that is critical for the induction of EMT in HCC cells. These data suggest a potential role for targeting FRAT1 in the prevention of hypoxia-induced HCC cancer progression and metastasis mediated by EMT.