E2F7/RAD51AP1 Axis Inhibits Endometrial Cancer Sensitivity to 5-FU via the Fatty Acid Metabolic Pathway.

BACKGROUND/AIM: Endometrial cancer (EC) is a frequent gynecological cancer. Studies have demonstrated that the sensitivity of EC toward 5-fluorouracil (5-FU) chemotherapy has decreased, leading to unsatisfactory treatment effects. There is an urgent need to investigate the reasons for the unsatisfactory treatment of EC with 5-FU. The purpose of the study was to investigate the effect of RAD51AP1 after being transcriptionally activated by E2F7 on the sensitivity of EC cells to 5-FU chemotherapy via the fatty acid metabolic pathway. MATERIALS AND METHODS: mRNA expression data on EC were downloaded from The Cancer Genome Atlas database, subjected to differential expression analysis, and the target genes were determined based on the bioinformatics analysis and literature consulting. The regulatory transcription factor upstream of RAD51AP1 in EC was predicted using the hTFtarget database. The expression of E2F7 and RAD51AP1 was measured by qRT-PCR and western blot. Then, the transcriptional activation relationship between E2F7 and RAD51AP1 was verified by chromatin immunoprecipitation (ChIP) and dual luciferase assays. The IC50 values of EC cells toward 5-FU were determined by the CCK-8 assay, and cell apoptosis was detected by flow cytometry. The expression of apoptosis-related and fatty acid metabolism-related proteins was evaluated by western blot. RESULTS: Bioinformatics analysis showed that both E2F7 and RAD51AP1 were highly expressed in EC, and the possible binding sites between RAD51AP1 promoter and E2F7 were predicted. ChIP assay and dual luciferase assay confirmed the binding of E2F7 to RAD51AP1 promoter region. Cell experiments showed that overexpressing RAD51AP1 could facilitate the growth and fatty acid metabolism of EC cells, and suppress cell sensitivity to 5-FU, while silencing of E2F7 could reduce the effect of RAD51AP1 overexpression on EC cell growth and sensitivity toward 5-FU. CONCLUSION: The E2F7/RAD51AP1 axis can promote the growth of EC cells and inhibit cell sensitivity to 5-FU by regulating fatty acid metabolism, suggesting that E2F7/RAD51AP1 axis may be a novel pathway for EC treatment.

CRABP2 regulates infiltration of cancer-associated fibroblasts and immune response in melanoma.

Finding biomarkers for immunotherapy is an urgent issue in cancer treatment. Cellular retinoic acid-binding protein 2 (CRABP2) is a controversial factor in the occurrence and development of human tumors. However, there is limited research on the relationship between CRABP2 and immunotherapy response. This study found that negative correlations of CRABP2 and immune checkpoint markers (PD-1, PD-L1, and CTLA-4) were observed in breast invasive carcinoma (BRCA), skin cutaneous melanoma (SKCM), stomach adenocarcinoma (STAD) and testicular germ cell tumors (TGCT). In particular, in SKCM patients who were treated with PD-1 inhibitors, high levels of CRABP2 predicted poor prognosis. Additionally, CRABP2 expression was elevated in cancer-associated fibroblasts (CAFs) at the single-cell level. The expression of CRABP2 was positively correlated with markers of CAFs, such as MFAP5, PDPN, ITGA11, PDGFRα/ß and THY1 in SKCM. To validate the tumor-promoting effect of CRABP2 in vivo, SKCM xenograft mice models with CRABP2 overexpression have been constructed. These models showed an increase in tumor weight and volume. Enrichment analysis indicated that CRABP2 may be involved in immune-related pathways of SKCM, such as extracellular matrix (ECM) receptor interaction and epithelial-mesenchymal transition (EMT). The study suggests that CRABP2 may regulate immunotherapy in SKCM patients by influencing infiltration of CAFs. In conclusion, this study provides new insights into the role of CRABP2 in immunotherapy response. The findings suggest that CRABP2 may be a promising biomarker for PD-1 inhibitors in SKCM patients. Further research is needed to confirm these findings and to explore the clinical implications of CRABP2 in immunotherapy.

Effects of oxidative stress caused by iron overload on arachidonic acid metabolites in MES23.5 cells.

Iron overload can induce oxidative stress, thereby inducing cell peroxidation. Arachidonic acid (ARA) is widely expressed in mammalian cells and esterified to membrane phospholipids. To explore the effect of iron overload on the metabolism of membrane phospholipids MES23.5 cells were treated with various concentrations of ferric ammonium citrate (FAC) to induce oxidative stress. Using UHPLC (I-Class LC, Waters) coupled to a QTRAP (AB Sciex 5500) technology, the contents of 13 substances of ARA and its metabolites were detected. When the cells were given two different concentrations of FAC, we found that both high and low concentrations decrease the expression of ARA (p=0.002, p=0.02) compared with the control group. ARA has three metabolic pathways: the COX pathway, LOX pathway and CYP450 pathway. Compared with the control group, the LTB4 content in the LOX pathway was decreased (p=0.10) after treatment with lowconcentration FAC, while the LTB4 content was increased in the high-concentration treatment group (p=0.06). However, the content of 12S-HETE (p=0.23, p=0.05) in the LOX metabolic pathway decreased with increase of FAC concentration. Similarly, the content of 15S-HETE also decreased with increase of FAC concentration (p=0.17, p=0.02). The other downstream metabolites of ARA, 9S-HODE (p=0.54, p=0.18) and 13S-HODE (p=0.81, p=0.13) were not significantly changed. The contents of thromboxane B2 (TXB2), leukotriene D4 (LTD4), prostaglandin E2 (PGE2), 8-iso-prostaglandin F2α (8-iso-PGF2α), prostaglandin F2α (PGF2α), 6-keto-PGF1α, and prostaglandin D2 (PGD2) were too low to be detected in MES23.5 cells. The above results indicate that oxidative stress caused by iron overload reduces the LOX metabolic pathway of ARA.

A new species and additional records of Lobrathium Mulsant amp; Rey (Coleoptera: Staphylinidae: Paederinae) from southern China.

Lobrathium chengzhifeii Lin Peng sp. n. (Yunnan: Mengsong) is described and illustrated. The female sexual characters of L. anatinum W.-R. Li and L.-Z. Li, 2013 are described and illustrated for the first time. Additional records of L. ablectum Assing, 2012, L. biaculeatum Assing, 2012, L. demptum Assing, 2012, L. diaoluoense W.-R. Li and L.-Z. Li, 2013, L. excisissimum Assing, 2012, L. hongkongense Bernhauer, 1931 and L. tortile F.-K. Zheng, 1988 are reported. Including the new taxa, 66 Lobrathium species are currently known from China.

InSexBase: an annotated genomic resource of sex chromosomes and sex-biased genes in insects.

Sex determination and the regulation of sexual dimorphism are among the most fascinating topics in modern biology. As the most species-rich group of sexually reproducing organisms on Earth, insects have multiple sex determination systems. Though sex chromosomes and sex-biased genes are well-studied in dozens of insects, their gene sequences are scattered in various databases. Moreover, a shortage of annotation hinders the deep mining of these data. Here, we collected the chromosome-level sex chromosome data of 49 insect species, including 34 X chromosomes, 15 Z chromosomes, 5 W chromosomes and 2 Y chromosomes. We also obtained Y-linked contigs of four insects species-Anopheles gambiae, Drosophila innubila, Drosophila yakuba and Tribolium castaneum. The unannotated chromosome-level sex chromosomes were annotated using a standard pipeline, yielding a total of 123 030 protein-coding genes, 2 159 427 repeat sequences, 894 miRNAs, 1574 rRNAs, 5105 tRNAs, 395 snoRNAs (small nucleolar RNA), 54 snRNAs (small nuclear RNA) and 5959 other ncRNAs (non-coding RNA). In addition, 36 781 sex-biased genes were identified by analyzing 62 RNA-seq (RNA sequencing) datasets. Together with 5707 sex-biased genes from the Drosophila genus collected from the Sex-Associated Gene Database, we obtained a total of 42 488 sex-biased genes from 13 insect species. All these data were deposited into InSexBase, a new user-friendly database of insect sex chromosomes and sex-biased genes. Database URL: http://www.insect-genome.com/Sexdb/.

Algorithms and Applications to Weighted Rank-one Binary Matrix Factorization.

Many applications use data that are better represented in the binary matrix form, such as click-stream data, market basket data, document-term data, user-permission data in access control, and others. Matrix factorization methods have been widely used tools for the analysis of high-dimensional data, as they automatically extract sparse and meaningful features from data vectors. However, existing matrix factorization methods do not work well for the binary data. One crucial limitation is interpretability, as many matrix factorization methods decompose an input matrix into matrices with fractional or even negative components, which are hard to interpret in many real settings. Some matrix factorization methods, like binary matrix factorization, do limit decomposed matrices to binary values. However, these models are not flexible to accommodate some data analysis tasks, like trading off summary size with quality and discriminating different types of approximation errors. To address those issues, this article presents weighted rank-one binary matrix factorization, which is to approximate a binary matrix by the product of two binary vectors, with parameters controlling different types of approximation errors. By systematically running weighted rank-one binary matrix factorization, one can effectively perform various binary data analysis tasks, like compression, clustering, and pattern discovery. Theoretical properties on weighted rank-one binary matrix factorization are investigated and its connection to problems in other research domains are examined. As weighted rank-one binary matrix factorization in general is NP-hard, efficient and effective algorithms are presented. Extensive studies on applications of weighted rank-one binary matrix factorization are also conducted.

Excessive tensile strain induced the change in chondrocyte phenotype.

PURPOSE: Chondrocyte extracellular matrix type II collagen and proteoglycans ensure an important compression-bearing structure in synovial joint. However, much more type I collagen is generated in osteoarthritis, which implies the presence of abnormal tensile strain in cartilage. We hypothesize that tensile stress influences chondrocyte phenotype at the cellular level, leading to potential osteoarthritis. METHODS: Chondrocytes were stimulated with cyclic excessive tensile (10%) or mild tensile or compressive strain (5%) at 0.5 Hz, 3 h per day for 3 days. Chondrocyte morphology and matrix proteoglycans level was separately determined by Rhodamine phalloidin and toluidine blue staining. The expression of cartilage marker molecules was measured using quantitative reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assays. RESULTS: Chondrocytes demonstrated significant fibroblastic morphology, reduced proliferation and increased apoptosis following exposure to 10% tensile strain. The 10% tensile strain group induced the lowest matrix proteoglycans level. It observably reduced the expression of COL2A1, Acan and SOX9, and increased COL1A1 expression level. The 5% tensile (5% compression) group, maintained the chondrocyte phenotype. CONCLUSIONS: The findings identified the effects of different magnitudes of tensile stress on chondrocyte phenotype compared to compressive strain. Further studies on cartilage biomechanical micro-environment might benefit from this study.

Morphological awareness as a function of semantics, phonology, and orthography and as a predictor of reading comprehension in Chinese.

This study investigates the contributions of semantic, phonological, and orthographic factors to morphological awareness of 413 Chinese-speaking students in Grades 2, 4, and 6, and its relationship with reading comprehension. Participants were orally presented with pairs of bimorphemic compounds and asked to judge whether the first morphemes of the words shared a meaning. Morpheme identity (same or different), whole-word semantic relatedness (high or low), orthography (same or different), and phonology (same or different) were manipulated. By Grade 6, children were able to focus on meaning similarities across morphemes while ignoring the distraction of form, but they remained influenced by whole-word semantic relatedness. Children's ability to overcome the distraction of phonology consistently improved with age, but did not reach ceiling, whereas the parallel ability for orthography reached ceiling at Grade 6. Morphological judgment performance was a significant unique predictor of reading comprehension when character naming and vocabulary knowledge were accounted for.

Preparation of a novel composite nanofiber gel-encapsulated human placental extract through layer-by-layer self-assembly.

Aqueous human placenta extract (HPE) has been previously used to treat chronic soft tissue ulcer; however, the optimal dosage of HPE has yet to be elucidated. The present study investigated a novel nanofiber gel composed through layer-by-layer (LbL) self-assembly, in which HPE was encapsulated. IKVAV, RGD, RAD16 and FGL-PA were screened and combined to produce an optimal vehicle nanofiber gel through LbL assembly. Subsequently, the aqueous HPE was encapsulated into this nanofiber at the appropriate concentration, and the morphology, particle size, drug loading efficacy, encapsulation rate, release efficiency and structure validation were detected. The encapsulation efficiency of all three HPE samples was >90%, the nanofiber gel exhibited a slow releasing profile, and the structure of HPE encapsulated in the nanofiber gel was unvaried. In conclusion, this type of novel composite nanocapsules may offer a promising delivery system for HPE.

Attenuation of oxygen fluctuation-induced endoplasmic reticulum stress in human lens epithelial cells.

Cataractogenic stresses are associated with the induction of endoplasmic reticulum (ER) stress. However, little is known about oxygen (O2)-induced ER stress in the lens. Cataract research has focused on elevated levels of O2 in lens epithelial cells (LECs). Excessive levels or a lack of O2 are known to induce ER stress whereas chronic ER stress activates the unfolded protein response (UPR). The present study investigated the hypothesis that the fluctuation of O2 levels induces a UPR, and may be controlled by maintaining human LECs (hLECs) in a specific concentration of O2. Human LECs were cultured in different atmospheric levels of O2. Hypoxic conditions were determined by the level of hypoxia-inducible factor (HIF)-1α. 2',7'-Dichlorodihydrofluorescein diacetate and ethidium homodimer-1 staining were conducted to detect reactive oxygen species (ROS) and cell death, respectively. Protein blot analyses were performed with antibodies specific to antioxidant and UPR-specific proteins. Reverse transcription-quantitatative polymerase chain reaction assays were performed to quantify the mRNA levels of activated NF-E2-related factor 2 (Nrf2) and kelch-like ECH-associated protein 1 (Keap1). The treatment of human LECs with 0 and 20% atmospheric O2 activated Nrf2/Keap1. The LECs shifted to 1% atmospheric O2 from 0, 4 or 20% for 24 h showed decreased levels of Keap1. By contrast, hLECs cultured in 1% atmospheric O2 for 24 h and then shifted to 0, 4 or 20% O2 exhibited a significant upregulation of Nrf2. These results suggest that oxidative stress proteins were not expressed in a 1% O2 environment. The O2 levels in the culture medium were equilibrated within 2 h in the cell culture plates. These results showed that an appropriate oxygen environment for the culture of LECs is ~1 % atmospheric O2. Either 0 or 20% of atmospheric O2 activated the UPR and the Nrf2/Keap1-mediated antioxidant system in LECs and chronic exposure to O2 fluctuation led to ROS production and cell death. This study revealed that O2 fluctuation-induced UPR/ER stress could be prevented by maintaining the cells in a 1% O2 environment.

Influence of Fetal Birth Weight on Caesarean Section Rate and Fetal Outcome After Induction of Labor.

AIM: The aim of the present study was to provide information for better obstetric counselling by analyzing the impact of fetal birth weight on the caesarean section rate and fetal outcome after induction of labor. MATERIALS AND METHODS: In this retrospective study from January 2010 to December 2013, 1,474 singleton deliveries with labor induction at or greater than 37 gestational weeks were analyzed for their impact of fetal birth weight on delivery outcome. The normal birth weight group was defined as 2,500 g to less than 4,000 g. For comparison, further birth weight groups were defined as: group 1 <2,500 g, group 2 4,000 to <4,250 g; group 3 ≥4,250 g. The primary outcome was the caesarean section rate; secondary outcome measures were fetal complications monitored by pH and base excess (BE) of the umbilical cord artery, Apgar score after 5 min (Apgar-5) and postpartum transfer to the Neonatal Care Unit. The set of controlling variables included maternal body mass index and age, gestational age, neonatal sex, maternal diabetes, maternal hypertension disorder, parity and method of induction of labor. RESULTS: Second-stage caesarean section is significantly more likely when fetal birth weight is below 2,500 g (42.9% vs. 24.2% in the normal birth weight group, odds ratio=3.11, 95% confidence interval=1.48-6.51, p=0.003). A birth weight of 4,000 g or more did not have a significant influence on the caesarean section rate. Only the mean Apgar-5 for group 1 was significantly lower (p=0.044). The non-parametric tests and regression analyzes of pH and BE of the umbilical cord and of the Apgar-5 for adverse fetal outcome (pH<7.05, BE<-12 or Apgar-5 <7) showed no significant differences in the three birth weight groups when compared to the normal group. Neonates were significantly more often transferred to the Neonatal Care Unit after delivery when birth weight was below 2,500 g (odds ratio=9.68, 95% confidence interval=4.33-21.65, p<0.001) or above 4,250 g (odds ratio=2.68, 95% confidence interval=1.34-5.36, p=0.005). CONCLUSION: Although a fetal birth weight of under 2500 g and a birth weight over 4,250 g are associated with some risks, there is no general contraindication against performing induction of labor in regards to fetal birth weight.

Serum levels of specific IgE to Staphylococcus aureus enterotoxins in patients with chronic rhinosinusitis.

The aim of this study was to determine the prevalence of Staphylococcus aureus enterotoxins (SEs) in the serum from patients with chronic rhinosinusitis (CRS) and its involvement in the condition. Thirty CRS patients without nasal polyps (CRSsNP), 40 CRS patients with nasal polyps (CRSwNP), and 30 healthy controls were enrolled in this study. Peripheral blood was obtained and analyzed to measure the serum levels of total IgE, specific IgE to SEA, SEB and SEC, and eosinophil cationic protein (ECP) using ImmunoCAP assays. The positive rate and level of serum specific IgE to SEB, but not to SEA or SEC, were significantly higher in CRSwNP patients compared with the controls (P=0.027 and P=0.021, respectively). No significant differences were found between CRSsNP patients and controls, or between CRSsNP and CRSwNP patients. Serum total IgE was significantly elevated and positively correlated with SEB-specific IgE in the CRSsNP (P<0.001; r=0.393, P=0.032) and CRSwNP (P<0.001; r=0.581, P<0.001) groups. ECP was also significantly increased in the CRSsNP (P=0.002) and CRSwNP (P<0.001) groups, but not correlated with specific IgE to SEs in either CRS group. The results suggest that SEB may play a role in the pathogenesis of CRSwNP.

MicroRNA-181a enhances the chemotherapeutic sensitivity of chronic myeloid leukemia to imatinib.

MicroRNA-181 (miR-181) has been recently demonstrated to participate in the differentiation and development of immune cells, including natural killer cells and B and T lymphocytes, and myeloid linages, including erythroid and megakaryocytic cells. The aberrant expression of miR-181, particularly low expression levels, has been observed in a number of leukemia types, including B-cell chronic lymphocytic leukemia and cytogenetically abnormal acute myeloid leukemia. However, the expression and function of miR-181 in chronic myeloid leukemia (CML) remains unknown. In the present study, the aberrant expression of miR-181a was analyzed in a patient with CML and in the CML K562 cell line. In addition, the function and potential mechanisms of miR-181a in K562 cells with regard to their chemotherapeutic sensitivity to imatinib were investigated. The expression levels of miR-181a were significantly reduced in the patient with CML and in the CML K562 cell line. Furthermore, the overexpression of miR-181a in the K562 cells enhanced the chemotherapeutic sensitivity of these cells to imatinib. The potential mechanism mediating these effects may be associated with the capacity of miR-181a to inhibit cell growth and/or to induce cells apoptosis and differentiation in K562 cells. The results of the present study suggested that miR-181a may be a target for the treatment of CML and a useful indicator of the therapeutic sensitivity of CML to imatinib.

Vascular expression of polycystin-2.

The expression of polycystin-1 in the vascular smooth muscle cells (VSMC) of elastic and large distributive arteries suggests that some vascular manifestations of autosomal-dominant polycystic kidney disease (ADPKD) result directly from the genetic defect. Intracranial aneurysms have been reported in PKD2, as well as in PKD1 families. To determine whether the vascular expression of polycystin-2 is similar to that of polycystin-1, the expression of PKD2 mRNA and protein in cultured pig aortic VSMC was studied and immunofluorescence and immunohistochemistry were used to study the localization of polycystin-2 in cultured pig aortic VSMC, pig ascending thoracic aorta, and normal elastic and intracranial arteries and intracranial aneurysms obtained at autopsy from patients without or with ADPKD. Tissues derived from Pkd2 wild-type and Pkd2 null mice were used to confirm the specificity of the immunostaining for polycystin-2. Northern blots of VSMC revealed the expected 5.3-kb band. Western blotting detected a 110-kb band in a 100,000 x g fraction of VSMC homogenates. Cultured VSMC as well as VSMC between the elastic lamellae of pig thoracic aorta were positive for polycystin-2 by immunofluorescence. The staining pattern was cytoplasmic. Treatment of the cells before fixation with Taxol, colchicine, or cytochalasin-D altered the pattern of staining in a way suggesting alignment with the cytoskeleton. The immunohistochemical staining for polycystin-2 was abolished by extraction with 0.5% Triton X-100, indicating that polycystin-2 is not associated with the cytoskeleton. Weak immunoreactivity for polycystin-2, which was markedly enhanced by protease digestion, was detected in formaldehyde-fixed normal human elastic and intracranial arteries. Immunostaining of variable intensity for polycystin-2, which was not consistently enhanced by protease digestion, was seen in the spindle-shaped cells of the wall of the intracranial aneurysms. The similar expression of polycystin-1 and polycystin-2 in the vascular smooth muscle is consistent with the proposed interaction of these proteins in a single pathway. These observations suggest a direct pathogenic role for PKD1 and PKD2 mutations in the vascular complications of ADPKD.