YY1/LncRNA GAS5 complex aggravates cerebral ischemia/reperfusion injury through enhancing neuronal glycolysis.

Yin-Yang 1 (YY1) has been identified as playing critical roles in multiple diseases. However, little is known regarding its roles and mechanisms in cerebral ischemia/reperfusion (I/R) injury. This study is aimed to explore the roles of YY1 in regulating neuronal apoptosis in cerebral I/R injury and its underlying mechanisms. Primary mouse cerebral cortical neurons were isolated and subjected to OGD/R to mimic cerebral I/R injury in vitro. The roles of YY1 on OGD/R-induced neuronal injury were investigated by performing western blotting, quantitative real-time polymerase chain reaction, TUNEL, RNA-binding protein immunoprecipitation, chromatin immunoprecipitation, chromatin isolation by RNA purification assay, glucose uptake assay, lactate production assay, and extracellular acidification rate assay. YY1-binding long non-coding RNAs (LncRNAs) in neurons subjected to OGD/R were identified by RIP and RNA sequencing. The roles of YY1 on cerebral I/R in vivo were detected by assessing neuronbehaviour, infarct size, and neuronal apoptosis. We found that YY1 expression is downregulated, and LncRNA GAS5 is upregulated in neurons subjected to OGD/R. OGD/R treatment promotes YY1 interacting with GAS5 in neurons, and YY1 negatively regulates GAS5 expression by binding to GAS5 promoter to repress its transcription. Besides, YY1 and GAS5 bind to the same region of PFKFB3 promoter to promote PFKFB3 expression and strengthen neuronal glycolysis, resulting in aggravating OGD/R-induced neuronal apoptosis. Knockdown of YY1 or GAS5 protects against I/R-induced ischemic brain damage and improves overall neurological functions in vivo. Overall, YY1 interacts with LncRNA GAS5 to promote PFKFB3 transcription to enhance neuronal glycolysis, resulting in aggravating cerebral I/R injury.

Pterygial body epithelium domination of pterygial proliferation with TCF4 as a potential key factor.

AIM: To characterize the proliferative capacity of pterygial epithelium in different regions (head, neck and body) of pterygium and explore the function of transcription factor 4 (TCF4) in pterygium proliferation. METHODS: Thirty pterygium tissues and 10 normal conjunctival tissues were obtained from Zhongshan Ophthalmic Center (ZOC) and Guangdong Eye Bank, respectively. Proliferative capacity of head, neck and body in pterygial epithelium was measured using clonal analysis, fold growth analysis and expression profile of proliferative markers revealed by immunofluorescent staining and real-time PCR. The expression of TCF4 was highlighted by double immunofluorescent staining with other proliferation related markers such as proliferating cell nuclear antigen (PCNA) and ATP-binding cassette sub-family G member 2 (ABCG2). RESULTS: The proliferative potential of pterygial epithelium was higher than that of normal conjunctival epithelium. High expression levels of proliferative markers (P63α, PCNA and ABCG2) in pterygial body epithelium were observed in immunofluorescent staining and real-time PCR (P<0.05). Also, epithelial cells isolated from pterygial body demonstrated higher proliferative capacity in clonal analysis and fold growth analysis, than those isolated from the head and neck regions. The TCF4 expression in pterygial epithelium was similar to other proliferative markers (P63α, PCNA and ABCG2), as higher in pterygial body than head and neck. Moreover, TCF4 showed coexpression with other proliferation-related markers (PCNA and ABCG2) in the double immunofluorescent staining experiment. CONCLUSION: The proliferative capacity in pterygial body epithelium is prominent than the head and neck regions, and upregulated TCF4 may be associated with enhanced proliferation in the pterygium.

The inflammation influence on corneal surface after frontalis suspension surgery.

AIM: To study the influence of frontalis muscle flap suspension on ocular surface by analyzing the clinical features and inflammatory cytokines. METHODS: A prospective, observational case series. Thirty-one eyes of 25 patients with severe congenital blepharoptosis who underwent frontalis muscle flap suspension surgery with at least 6mo of follow-up were included in the study. The main outcome measures were margin reflex distance 1 (MRD1), degree of lagophthalmos, ocular surface disease index (OSDI), fluorescein staining (Fl), tear break-up time (BUT), Schirmer I test, and inflammatory cytokine assay. RESULTS: The degrees of lagophthalmos significantly increased after surgery. The OSDI scores significantly increased 1wk postoperatively and then decreased 4wk after operation. The Fl scores reflected corneal epithelial defects in sixteen patients at early stage postoperatively. The BUT and Schirmer I test values remained stable and did not show change compared to those before surgery. The inflammatory cytokines in conjunctival epithelial cells (including IL-1ß, IL-6, IL-8, TNF-α, and IL-17A) significantly increased 1wk after the surgery (P<0.001), then returned to the normal level at 24wk postoperatively. The levels of inflammatory cytokine IL-1ß, IL-6, IL-8, TNF-α, and IL-17A elevated significantly and were positively correlated with OSDI and Fl scores. CONCLUSION: Frontalis muscle flap suspension surgery results in lagophthalmos in early period of post-operation and relieved after months. The elevation of inflammatory cytokines level may participate in the occurrence of corneal epithelial defects at the early postoperative stage.

YY1 promotes IL-6 expression in LPS-stimulated BV2 microglial cells by interacting with p65 to promote transcriptional activation of IL-6.

Neuroinflammation plays a critical role in the process of neurodegenerative disorders, during which microglia, the principal resident immune cells in the central nervous system, are activated and produce proinflammatory mediators. Yin-Yang 1 (YY1), a multi-functional transcription factor, is widely expressed in cells of the immune system and participate in various cellular processes. However, whether YY1 is involved in the process of neuroinflammation is still unknown. In the present study, we found that YY1 was progressively up-regulated in BV2 microglial cells stimulated with lipopolysaccharide (LPS), which was dependent on the transactivation function of nuclear factor kappa B (NF-κB). Furthermore, YY1 knockdown notably inhibited LPS-induced the activation of NF-κB signaling and interleukin-6 (IL-6) expression in BV-2 cells, but not mitogen-activated protein kinase (MAPK) signaling. Moreover, YY1 strengthened p65 binding to IL-6 promoter by interacting with p65 but decreased H3K27ac modification on IL-6 promoter, eventually increasing IL-6 transcription. Taken together, these results for the first time uncover the regulatory mechanism of YY1 on IL-6 expression during neuroinflammation responses and provide new lights into neuroinflammation.

Leptomeningeal carcinomatosis as the initial manifestation of gastric adenocarcinoma: a case report.

Leptomeningeal involvement is usually reported as a secondary event in advanced gastric carcinoma. Leptomeningeal carcinomatosis (LMC), as the initial manifestation of asymptomatic gastric cancer, is exceedingly rare with only a few cases reported in recent years. The presenting neurologic symptoms include headache, vomiting and seizures and are usually clinically atypical. The diagnosis of LMC is made via identification of malignant cells that originate from epithelial cells in the cerebrospinal fluid by cytological examination and provides cues to track the primary tumor. Endoscopic examinations are crucial to confirm the presence of gastric cancer, and imaging studies, especially gadolinium-enhanced magnetic resonance imaging of the brain, are sometimes helpful in diagnosis. Thus far, there is no standard therapy for LMC, and despite all measures, the prognosis of the condition is extremely poor. Here, we report on the clinical features and diagnostic procedures for a patient with occult gastric cancer with Bormann type I macroscopic appearance and poor differentiation in pathology, who presented with LMC-induced neurological symptoms as the initial clinical manifestation. Additionally, we review the similar cases reported over the past years, making comparison among cases in order to provide more information for the future diagnosis.

Molecular characterization of a defense-related AMP-binding protein gene, OsBIABP1, from rice.

We cloned and characterized a rice gene OsBIABP1 encoding an AMP-binding protein. The full-length cDNA of OsBIABP1 is 1912-bp long and is predicted to encode a 558-aa protein. OsBIABP1 contains a typical AMP-binding signature motif and shows high similarity to members of AMP-binding protein family. OsBIABP1 is expressed in stems, leaves and flowers of rice plants, but is not expressed, or expressed at a very low level, in rice roots. The expression of OsBIABP1 was induced by some defense-related signal molecules, e.g., salicylic acid (SA), benzothiadiazole, jasmonic acid (JA), and 1-amino cyclopropane-1-carboxylic acid, which mediate SA- and JA/ethylene (ET)-dependent defense signaling pathways, respectively. Furthermore, the expression of OsBIABP1 is activated by the infection of Magnaporthe oryzae, and the induced expression is quicker and stronger during early stages of pathogenesis in incompatible interaction than that in compatible interaction between rice and M. oryzae. Our results suggest that OsBIABP1 may be a defense-related AMP-binding protein that is involved in the regulation of defense response through SA and/or JA/ET signaling pathways.

[Validation of the non-cellular metastasis hypothesis of malignant tumors in mice].

OBJECTIVE: To test the hypothesis that malignant tumor metastasis is mediated also through a non-cellular, essentially molecular, mechanism in addition to the cellular pathway. METHODS: The sex-determining region on the Y chromosome was detected as the marker of the primary tumors using PCR in Lewis lung carcinoma (LLC) in vitro and in female C57BL/6 mice bearing LLC with spontaneous metastasis. The macroscopic and microscopic metastases in the tumor-bearing mice were examined for SRY expression by PCR and in situ hybridization, using the tissues from male and female mice as the positive or negative controls. RESULTS AND CONCLUSION: Positive SRY gene expression was detected in the metastatic foci in the LLC-bearing female mice, suggesting the origination of these tumor cells from the primary tumor foci. We have failed to verify the non-cellular metastasis hypothesis in this animal experiment, but given the limitations of this experiment, we consider further investigation still necessary for verification of this hypothesis using other methods.

[Dynamic finite element analysis of three ferrule designs in post crown under impact loading].

OBJECTIVE: To investigate the stress distribution in three ferrule designs in a maxillary central incisor restoration using PFM crown with post, and evaluate the biomechanical mechanism of the ferrule effect in the post crown by 3D finite element dynamic analysis. METHODS: The 3D finite element model of a maxillary central incisor restored with post and PFM crown was established. By simulating three types of ferrule effect [crown wrapping dentine (A), core collar wrapping (B), and contrabevel (C)] under dynamic loading, the dentin stress was analyzed. RESULTS: During dynamic loading, the stress distribution tended to increase from the cervical part to root middle and radical part of the tooth, and greater high stress area was found around the apex of the post, where the peak stress was observed value. The stress of the labial dentin of the root inferior segment increased obviously. The high stress areas were invariable at every loading step during dynamic loading. The peak stress was sigma(vonA)

[A three dimensional finite element study on stress distribution in maxillary central incisor restored with fiber post].

OBJECTIVE: To study the stress changes of maxillary central incisor restored with or without fiber post using three dimensional finite element method, and analysis the role of fiber post in determining the stress distribution in dentin. METHODS: Three dimensional finite element models of maxillary central incisor with various remaining tooth structure were established by spiral CT, Mimics software and ANSYS software. Test samples were restored with all-ceramic crown and fiber post all-ceramic crown, respectively. The von Mises stress and maximal tensile stress of dentin were recorded. RESULTS: The stress level in dentin of maxillary central incisor restored with fiber post all-ceramic crown was smaller than that restored with all-ceramic crown, the stress distribution of both were similar. CONCLUSION: The apply of fiber post can reduce the stress level in dentin of maxillary central incisor and decrease the risk of tooth breakage, but not change the stress pattern.

FISSION1A and FISSION1B proteins mediate the fission of peroxisomes and mitochondria in Arabidopsis.

Peroxisomes and mitochondria are metabolically diverse organelles that act in concert in a number of pathways in eukaryotes, including photorespiration and lipid mobilization in plants. The division machineries of these two types of organelles also share several components such as dynamin-related proteins (DRPs) and their organelle anchor, the FISSION1 (FIS1) protein. In Arabidopsis, members of the DRP3 and FIS1 small protein families, namely DRP3A, DRP3B, FIS1A, and FIS1B, are each dual-targeted to peroxisomes and mitochondria and are required for the division of both organelles; DRP3A and DRP3B are partially redundant in function. To further determine the contribution of FIS1A and FIS1B to the division of peroxisomes and mitochondria, we analyzed plants overexpressing FIS1A or FIS1B and mutants in which the functions of both proteins were disrupted. Domains in FIS1A and FIS1B required for peroxisomal targeting were also dissected. Our results demonstrate that FIS1A and FIS1B play rate-limiting and partially overlapping roles in promoting the fission of peroxisomes and mitochondria. Furthermore, although the C-terminus of FIS1 is both necessary and sufficient for targeting to peroxisomes, the role of the short C-terminal segment adjacent to the transmembrane domain may differ among diverse species in peroxisomal targeting.

[Comparison of the failure mode of various types of glass ionomer cements].

OBJECTIVE: To investigate the failure mode of various types of glass ionomer cements by Hertzian indentation test. METHODS: Discs of 10 mm diameter and 2 mm thickness were prepared for six glass ionomer cement products (A-D: conventional type setting through an acid-base chemical reaction, A and B without reinforcement, C with silver reinforcement, D with ceramic reinforcement; E and F: resin-modified type), ten for each. These were tested on top of glass-reinforced polyamide-nylon 6, 6 substrates by a universal testing machine, loading centrally with a 20 mm diameter ball. Load at the first crack was recorded. Failure mode was observed under scanning electron microscope. RESULTS: The former four products presented typical brittle fracture, while the latter two usually fractured incompletely. The failure loads at the first crack of the six glass ionomer cements were (258.86 +/- 10.49), (230.88 +/- 21.66), (281.90 +/- 25.39), (282.11 +/- 9.60), (756.67 +/- 83.50) and (1 148.00 +/- 147.78) N, respectively. Significant difference was found between the former four and the latter two products. CONCLUSIONS: The type (setting mode) of glass ionomer cement controls its failure mode. Inclusion of metallic or ceramic filler has little effect on increasing the load bearing capacity of glass ionomer cement.

[Effect of staining method and sintering temperature on the color of porcelain-fused-to-metal restorations].

PURPOSE: To investigate the effect of staining method and sintering temperature on the color of porcelain-fused-metal restorations. METHODS: 40 cylindrical stained porcelain-metal specimens of 15 mm in diameter and 6 mm in height were fabricated with customized mould, consisting of 2 mm Ni-Cr metal, 1 mm opaque porcelain, 2 mm dentine porcelain and 1 mm enamel porcelain. The specimens were prepared by 5 techniques, 8 for each group. Group A: internal staining, Group B: external staining, 900 degrees centigrade sintering temperature was used in both A and B group; Group C to E: external staining, with the sintering temperature of 880 degrees centigrade, 900 degrees centigrade and 920 degrees centigrade respectively. Sofu A2 porcelain and Sofu 44 stain system were used for the study. Using standard white plate as a reference, colors (L*,a*,b* coordinates) of the specimens were measured with a computerized colorimeter. Student's t test and one way ANOVA were used to analyze the data. RESULTS: DeltaE, b* and Delta C(ab)* of Group B (external staining) and group A (internal staining) were 43.72 +/- 2.99/26.51 +/- 1.64/31.31 +/- 2.48 and 39.71 +/- 1.78/23.69 +/- 0.36/26.55 +/- 2.16, respectively. The values of the former group were significantly higher than that of the latter (P < 0.05); For Group C to E, there were no significant differences in all the color parameters. CONCLUSIONS: Staining method has a significant effect on the color parameters of porcelain-fused-to-metal restorations; For external staining, within the clinically-used range, changing the sintering temperature does not have an obvious effect on the color.