Clinical significance of CTGF and Cry61 protein in extraocular muscles of strabismic patients.

PURPOSE: To investigate the relationship between clinical features and protein amounts of Cysteine-rich 61 (Cyr61/CCN1) and connective tissue growth factor (CTGF/CCN2), which are vital components and regulators of the extracellular matrix in resected muscles from strabismus surgery. METHODS: Strabismus patients who were diagnosed with horizontal concomitant strabismus or inferior oblique overaction (IOOA) and required extraocular muscles (EOMs) resection to correct eye position were included in this study. The protein amounts were measured by enzyme-linked immunosorbent assay (ELISA) in resected EOMs. Multivariable linear regression was used to investigate the associations, adjusting for gender, age (continuous), amblyopia, and disease duration. RESULTS: A total of 141 muscles (including 38 lateral, 81 medial rectus, and 22 inferior oblique muscles) from 128 patients were collected in this study. The amount of Cry61 and CTGF per millimeter was significantly negatively associated with deviation angle in intermittent exotropia patients (Cry61: ß, - 1.44; 95%CI, - 2.79 to - 0.10, p = 0.035; CTGF: ß, - 3.14; 95%CI, - 5.06 to - 1.22, p = 0.002). The same relationship was also detected in the partially accommodative and non-accommodative esotropia patients, although it was not statistically significant (Cry61: ß, - 2.40; 95%CI, - 5.05 to 0.24; p = 0.073; CTGF: ß, - 3.47; 95%CI, - 9.18 to 2.87; p = 0.269). The amount of Cry61 and CTGF per millimeter showed significant associations with the degree of IOOA (p < 0.05). CONCLUSIONS: Taken together, our results demonstrated a significant relationship between deviation angle and protein amount of Cry61 and CTGF and implied that Cry61 and CTGF may play important roles in modulation of EOM contractility, which provide new insights into strabismus pathogenesis.

Rapid Detection of blaKPC in Carbapenem-Resistant Enterobacterales Based on CRISPR/Cas13a.

Klebsiella pneumoniae carbapenemase (KPC) is a crucial enzyme that causes carbapenem resistance in Enterobacterales, and infections by these "superbugs" are extremely challenging to treat. Therefore, there is a pressing need for a rapid and accurate KPC detection test to control the prevalence of carbapenem-resistant Enterobacterales (CREs). In this study, we established a novel method for detection of blaKPC, the gene responsible for encoding KPC, based on a recombinase polymerase amplification (RPA) and a CRISPR/Cas13a reaction coupled to fluorophore activation (termed RPA-Cas13a assay). We carefully selected a pair of optimal amplification primers for blaKPC and achieved a lower limit of detection of approximately 2.5 copies/µL by repeatedly amplifying a recombinant plasmid containing blaKPC. The RPA-Cas13a assay demonstrated a sensitivity of 96.5% and specificity of 100% when tested on 57 blaKPC-positive CRE strains, which were confirmed by DNA sequencing. Moreover, in 311 sputum samples, the theoretical antibiotic resistance characteristics of blaKPC-positive strains obtained by the RPA-Cas13a assay were highly consistent with the results of antibiotic susceptibility test (Kappa = 0.978 > 0.81, P < 0.01). In conclusion, the RPA-Cas13a system is a simple and one-hour efficient technology for the detection of a potentially fatal antibiotic resistance gene.

Iron-Deficiency and Estrogen Are Associated With Ischemic Stroke by Up-Regulating Transferrin to Induce Hypercoagulability.

RATIONALE: Epidemiological studies have identified an associate between iron deficiency (ID) and the use of oral contraceptives (CC) and ischemic stroke (IS). To date, however, the underlying mechanism remains poorly understood. Both ID and CC have been demonstrated to upregulate the level and iron-binding ability of Tf (transferrin), with our recent study showing that this upregulation can induce hypercoagulability by potentiating FXIIa/thrombin and blocking antithrombin-coagulation proteases interactions. OBJECTIVE: To investigate whether Tf mediates IS associated with ID or CC and the underlying mechanisms. METHODS AND RESULTS: Tf levels were assayed in the plasma of IS patients with a history of ID anemia, ID anemia patients, venous thromboembolism patients using CC, and ID mice, and in the cerebrospinal fluid of some IS patients. Effects of ID and estrogen administration on Tf expression and coagulability and the underlying mechanisms were studied in vivo and in vitro. High levels of Tf and Tf-thrombin/FXIIa complexes were found in patients and ID mice. Both ID and estrogen upregulated Tf through hypoxia and estrogen response elements located in the Tf gene enhancer and promoter regions, respectively. In addition, ID, administration of exogenous Tf or estrogen, and Tf overexpression promoted platelet-based thrombin generation and hypercoagulability and thus aggravated IS. In contrast, anti-Tf antibodies, Tf knockdown, and peptide inhibitors of Tf-thrombin/FXIIa interaction exerted anti-IS effects in vivo. CONCLUSIONS: Our findings revealed that certain factors (ie, ID and CC) upregulating Tf are risk factors of thromboembolic diseases decipher a previously unrecognized mechanistic association among ID, CC, and IS and provide a novel strategy for the development of anti-IS medicine by interfering with Tf-thrombin/FXIIa interactions.

Clinical efficacy of conbercept plus micropulse laser (577 nm) treatment in macular edema secondary to non-ischemic central retinal vein occlusion.

Objectives: To observe the clinical efficacy and complications of conbercept combined with a 577-nm micropulse laser in patients with macular edema (ME) secondary to non-ischemic central retinal vein occlusion (CRVO). Methods: A total of 153 patients who were diagnosed with non-ischemic CRVO-induced ME and treated by Shijiazhuang People's Hospital during January 2019 and January 2021 were included in this study. The patients were divided into a control group and an experimental group by choice of treatment. The control group was treated by conbercept alone, while the experimental group underwent laser treatment on this basis. The best corrected visual acuity (BCVA) was determined by the internationally standardized logarithm of the minimum angle of resolution (logMAR) chart and the central macular thickness (CMT) was measured by optical coherence tomography (OCT) before and at one and three months after treatment. Complications such as conjunctival hemorrhage and elevated intraocular pressure (if any) were recorded during the 3-month follow-up period. Results: Conbercept could improve the BCVA and CMT of patients with non-ischemic CRVO (P <0.05, respectively), and the combined use of conbercept with micropulse laser treatment yielded greater improvements in these measures compared with the pre-treatment condition (P <0.05, respectively); moreover, the differences between the control group and the experimental group were statistically significant (all P <0.05). The two groups had similar complication rates and did not record any serious adverse reactions. Conclusions: Conbercept combined with 577-nm micropulse laser treatment can benefit patients with non-ischemic CRVO-induced ME by improving their visual acuity and relieve ME symptoms. As a regimen of impressive clinical efficacy, it is of great value for wide application in clinical practice.

Diboron(4)-Catalyzed Remote [3+2] Cycloaddition of Cyclopropanes via Dearomative/Rearomative Radical Transmission through Pyridine.

Ring structures such as pyridine, cyclopentane or their combinations are important motifs in bioactive molecules. In contrast to previous cycloaddition reactions that necessitated a directly bonded initiating functional group, this work demonstrated a novel through-(hetero)arene radical transmission concept for selective activation of a remote bond. An efficient, metal-free and atom-economical [3+2] cycloaddition between 4-pyridinyl cyclopropanes and alkenes or alkynes has been developed for modular synthesis of pyridine-substituted cyclopentanes, cyclopentenes and bicyclo[2.1.1]hexanes that are difficult to access using known methods. This complexity-building reaction was catalyzed by a very simple and inexpensive diboron(4) compound and took place via dearomative/rearomative processes. The substrate scope was broad and more than 100 new compounds were prepared in generally high yields. Mechanistic experiments and density function theory (DFT) investigation supported a radical relay catalytic cycle involving alkylidene dihydropyridine radical intermediates and boronyl radical transfer.

A Study of Differences in Penile Dorsal Nerve Somatosensory Evoked Potential Testing Among Healthy Controls and Patients With Primary and Secondary Premature Ejaculation.

BACKGROUND: Penile dorsal nerve somatosensory evoked potential (DNSEP) is a scientific and objective technique that provides effective and objective data to establish the diagnosis of premature ejaculation (PE). AIM: To explore differences in DNSEP between patients with primary premature ejaculation (PPE) and those with secondary premature ejaculation (SPE), in order to investigate the clinical value of DNSEP in the diagnosis of PE. METHODS: The participants were divided into a PPE group (34 cases), an SPE group (25 cases) and a healthy control group (18 cases). All participants underwent DNSEP testing, and the latencies and amplitudes of DNSEP were recorded. OUTCOMES: Differences in the latencies and amplitudes of DNSEP were compared among the PPE, SPE, and healthy control groups. RESULTS: The latencies of DNSEP in the PPE and SPE groups were shorter than those in the healthy control group, and these differences were statistically significant (P < 0.01). However, there was no statistically significant difference between the PPE and SPE groups (P > 0.05). The amplitudes of DNSEP in the PPE group were significantly higher than those in the healthy control group (P < 0.01). However, the amplitudes of DNSEP in the SPE group were significantly lower than those in the healthy control group (P < 0.05). CLINICAL IMPLICATIONS: PPE and SPE can be differentiated based on differences in the amplitudes of DNSEP, providing an objective basis for treatments and follow-up examinations. STRENGTHS AND LIMITATIONS: We evaluated differences in the amplitudes of DNSEP between PPE and SPE patients, which were rare in the published literature. However, specific causes of these differences are still unclear. SEP only reflects afferent pathways in the ejaculatory reflex arc, and role of the brain as a higher center should not be ignored. CONCLUSION: Both PPE and SPE patients are characterized by an increased excitability of the penile sensory nerves. Sun Z, Liao Z, Zheng Q, et al. A Study of Differences in Penile Dorsal Nerve Somatosensory Evoked Potential Testing Among Healthy Controls and Patients With Primary and Secondary Premature Ejaculation. J Sex Med Rev 2021;18:732-736.

Analysis of choroidal thickness in patients with proliferative diabetic retinopathy by optical coherence tomography angiography.

OBJECTIVES: This study aimed to evaluate the changes in choroidal thickness at different parts in patients with stage IV-V diabetic retinopathy (DR) treated by Panretinal Photocoagulation (PRP) or its combination with anti-vascular endothelial growth factor (VEGF) therapy using optical coherence tomography angiography (OCTA). METHODS: Patients with proliferative DR (stage IV-V) diagnosed in Shijiazhuang People's Hospital between January 2016 to January 2020 were selected and treated with conventional PRP or combined with anti-VEGF therapy. OCTA was performed before treatment and one three and six months after treatment to observe and compare subfoveal choroidal thickness (SFCT), perifoveal choroidal thickness at 500 um (M500) and 1500 um (M1500). RESULTS: A total of 76 patients (133 eyes) were included. Six months later, re-examination showed effective treatment in 122 eyes (91.72%) and ineffective treatment in 11 eyes. Before treatment and one week, three months and six months after treatment, the choroidal thickness was observed and compared by OCTA. SFCT, M500 and M1500 increased one week after treatment, were significantly thinner 3 months after treatment than those before treatment, and further decreased six months after treatment. CONCLUSION: OCTA presents a good evaluation of perifoveal choroidal thickness in patients with proliferative DR. It provides a basis for treatment selection and efficacy determination of proliferative DR.

Altered inhibitory synapses in de novo GABRA5 and GABRA1 mutations associated with early onset epileptic encephalopathies.

We performed next generation sequencing on 1696 patients with epilepsy and intellectual disability using a gene panel with 480 epilepsy-related genes including all GABAA receptor subunit genes (GABRs), and we identified six de novo GABR mutations, two novel GABRA5 mutations (c.880G>T, p.V294F and c.1238C>T, p.S413F), two novel GABRA1 mutations (c.778C>T, p.P260S and c.887T>C, p.L296S/c.944G>T, p.W315L) and two known GABRA1 mutations (c.335G>A, p.R112Q and c.343A>G, p.N115D) in six patients with intractable early onset epileptic encephalopathy. The α5(V294F and S413F) and α1(P260S and L296S/W315L) subunit residue substitutions were all in transmembrane domains, while the α1(R112Q and N115R) subunit residue substitutions were in the N-terminal GABA binding domain. Using multidisciplinary approaches, we compared effects of mutant GABAA receptor α5 and α1 subunits on the properties of recombinant α5ß3γ2 and α1ß3γ2 GABAA receptors in both neuronal and non-neuronal cells and characterized their effects on receptor clustering, biogenesis and channel function. GABAA receptors containing mutant α5 and α1 subunits all had reduced cell surface and total cell expression with altered endoplasmic reticulum processing, impaired synaptic clustering, reduced GABAA receptor function and decreased GABA binding potency. Our study identified GABRA5 as a causative gene for early onset epileptic encephalopathy and expands the mutant GABRA1 phenotypic spectrum, supporting growing evidence that defects in GABAergic neurotransmission contribute to early onset epileptic encephalopathy phenotypes.

Functional acute acquired comitant esotropia: clinical characteristics and efficacy of single Botulinum toxin type A injection.

BACKGROUND: To examine the clinical features of acute acquired comitant esotropia (AACE) and to evaluate the clinical effectiveness of a single injection of botulinum toxin type A (BTXA) on binocular visual function (BVF). METHODS: This retrospective, observational case series study enrolled patients with AACE examined from October 2018-May 2019. BTXA was injected into the both medial rectus muscles. The refractive error, best-corrected visual acuity (BCVA), stereoacuity, vergence, accommodation, the horizontal angle of deviation, and the gradient accommodative convergence/accommodation (AC/A) ratio were measured pre- and post-BTXA injection. Data pre- and postinjection were compared by the Wilcoxon signed-rank test. A Spearman correlation coefficient was calculated to explore the relationships between demographic characteristics and BVF. RESULTS: Twenty-two AACE cases were included. Compared with preinjection deviation, the postinjection deviation in the primary position was smaller for near (p < 0.001) and distance (p < 0.001) fixation at 3 months after injection (BTXA). Furthermore, convergence was better for near (p = 0.003) and distance (p < 0.001) fixation, divergence was better for near (p = 0.021) and distance (p < 0.001) fixation, accommodation was better in the right (p = 0.011) and left (p = 0.004) eyes, and the gradient AC/A ratio was better at the third month after injection (p = 0.001). Stereoacuity was improved in 11 (50%), unchanged in 5 (22.73%) and decreased in 6 (27.27%) patients. The preinjection stereoacuity (p = 0.013, r = 0.522) and preinjection deviation for near (p = 0.015 r, = - 0.512) and distance (p = 0.009, r = - 0.541) were significantly associated with patient age. CONCLUSIONS: AACE is characterized by a high AC/A ratio and low accommodation. A single injection of BTXA is effective for AACE. Deviation, stereoacuity, and the therapeutic effect of BTXA may be correlated with patient age.

The effect of spleen tyrosine kinase inhibitor R406 on diabetic retinopathy in experimental diabetic rats.

PURPOSE: To investigate the effect of spleen tyrosine kinase (Syk) inhibitor R406 on diabetic retinopathy (DR) in diabetic mellitus (DM) rats. METHODS: Rats were randomized into Normal, DM, DM + 5 mg/kg R406 and DM + 10 mg/kg R406 groups. DM rats were established via injection of streptozotocin (STZ). One week after model establishment, rats in treatment groups received 5 mg/kg or 10 mg/kg R406 by gavage administration for 12 weeks consecutively, followed by the detection with hematoxylin-eosin (HE) staining, Evans blue angiography, retinal trypsin digestion assay, Western blotting, immunohistochemistry, TUNEL assay, immunofluorescence assay and quantitative reverse transcriptase real-time polymerase chain reaction (qRT-PCR). RESULTS: The retina of DM rats presented different degree of edema, disordered and loose structure, swollen cells with enlarged intercellular space, and dilated and congested capillaries. Besides, the retinal vessels of DM rats showed high fluorescence leakage. However, R406 alleviated the above-mentioned conditions, which was much better with high concentration of R406 (10 mg/kg). R406 also reversed the down-regulations of occludin, claudin-5, ZO-1 and the up-regulation of and VEGF in retinal tissues of DM rats; inhibited retinal cell apoptosis; strengthened retinal cell proliferation; and reduced expressions of IL-1ß, IL-6, TNF-α and nuclear p65 NF-κB in retinal tissues. The improvement in all these indexes was much more significant in rats of DM + 10 mg/kg R406 group than in rats of DM + 5 mg/kg R406 group. CONCLUSION: Syk inhibitor R406 could attenuate retinal inflammation in DR rats via the repression of NF-κB activation.

Glutamate Ionotropic Receptor Kainate Type Subunit 3 (GRIK3) promotes epithelial-mesenchymal transition in breast cancer cells by regulating SPDEF/CDH1 signaling.

Glutamate Ionotropic Receptor Kainate Type Subunit 3 (GRIK3) is an important excitatory neurotransmitter receptor that plays a significant role in various neurodegenerative diseases. However, the biological functions of GRIK3 in malignancies are largely unknown because of limited related studies. Here, we primarily reported that the expression of GRIK3 was higher in breast cancer tissues than in adjacent noncancerous tissues. GRIK3 expression was also positively correlated with the prognosis of patients with breast cancer. GRIK3 promoted the proliferation and migration abilities of breast cancer cells and enhanced the growth of orthotopically implanted tumors. Mechanically, GRIK3 influenced a range of signaling pathways and key signal transducers, including two epithelial-mesenchymal transition regulators, SPDEF and CDH1. Heterogenous expression of SPDEF and CDH1 counteracted the migration and invasion abilities, respectively, of breast cancer cells induced by GRIK3. Moreover, overexpression of GRIK3 increased the expression of mesenchymal markers and decreased the expression of epithelial markers, resulting in the translocation of ß-catenin into the nucleus and the increased ß-catenin transcriptional activity. In conclusion, the present study reported a novel oncogenic role of GRIK3. Meanwhile, GRIK3, as a membrane receptor, may also serve as a potential therapeutic target for the treatment of breast cancer.

microRNA-mRNA analysis in pituitary and hypothalamus of sterile Japanese flounder.

Double haploidy is an advantageous situation for genetic mapping and genome sequencing studies. In the present study, the hypothalamus and pituitary gland from sterile and fertile double-haploid (DH) Japanese flounders (aged 5 years) were used as experimental materials for studying the expression of genes in individuals with reproductive disorders, using high-throughput sequencing technology. The results revealed abnormal levels of some hormones in sterile DHs during the breeding season. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that the significantly different microRNAs and messenger RNAs were related to metabolism, signal transduction, and melanogenesis; those related to steroid hormone synthesis and secretion related pathways were not detected. Our results suggest that the key to sterility in DHs was the arrested ovary development. However, the reason for arrested ovary development was mainly related to the lower levels of expression of genes involved in steroid biosynthesis in gonads, and was not related to the pituitary. For maintaining homeostasis, the hypothalamus and pituitary would have large differences in several processes, including signal transduction, metabolism, and immune response. The present study provides primary data for further studies on sterility in fish, and even in other animals.

Glutamate metabotropic receptor 4 (GRM4) inhibits cell proliferation, migration and invasion in breast cancer and is regulated by miR-328-3p and miR-370-3p.

BACKGROUND: Glutamate metabotropic receptors (GRM) play a variety of roles in neuronal cells. However, their clinical significance and biological functions in breast cancer remain unknown. METHODS: RNA sequencing data of breast cancer was obtained from the TCGA dataset (v2) and mined for the expression profiles of GRM family according to cancer subtypes. mRNA expression of GRM family in breast cancer tissues and para-cancerous tissue samples as well as breast cancer cell lines were measured by qPCR. The effects of over- and under-expression of GRM4 on cell capabilities to survive, migrate and invade were determined by colony formation, transwell migration and invasion assays. To explore the upstream regulation pattern of GRM4, miRNAs that target GRM4 were predicted and validated by dual luciferase reporter assay. In addition, the mRNA and protein expression of GRM4 regulated by these miRNAs were further measured by qPCR and western blot assay. RESULTS: GRM4 was the only GRM member that expressed in breast cancer tissues. Ectopic expression of GRM4 was correlated with better prognosis of breast cancer patients. Overexpression of GRM4 could significantly inhibit cell proliferation, migration and invasion capacity in MDA-MB-231, while knockdown of GRM4 could promote these processes. miR-328-3p and miR-370-3p were predicted to regulate the expression of GRM4 and dual luciferase reporter assay demonstrated that miR-328-3p and miR-370-3p directly bound to the 3' UTR of GRM4 and mutations on the binding regions on GRM4 significantly decreased the luciferase activity. qPCR demonstrated that expression of miR-328-3p and miR-370-3p was significantly decreased in breast cancer tissues and cells compared with that in control samples. However, there were no correlations between the expression of miR-328-3p and GRM4, as well as the expression of miR-370-3p and GRM4. Moreover, overexpression of miR-328-3p and miR-370-3p counteracted the inhibitory effect of GRM4-induced cell proliferation, migration and invasion. CONCLUSIONS: Our results suggest that GRM4 might be a tumor suppressor gene in breast cancer under the direct regulation of miR-328-3p and miR-370-3p.

Identification of key genes relevant to the prognosis of ER-positive and ER-negative breast cancer based on a prognostic prediction system.

Few prognostic indicators with differential expression have been reported among the differing ER statuses. We aimed to screen important breast cancer prognostic genes related to ER status and to construct an efficient prognostic prediction system. mRNA expression profiles were downloaded from TCGA and GSE70947 dataset. Two hundred seventy-one overlapping differentially expressed genes (DEGs) between the ER- and ER+ breast cancer samples were identified. Among the 271 DEGs, 109 prognostically relevant mRNAs were screened. mRNAs such as RASEF, ITM2C, CPEB2, ESR1, ANXA9, and VASN correlated strongly with breast cancer prognosis. Three modules, which contained 28, 9 and 8 enriched DEGs, were obtained from the network, and the DEGs in these modules were enriched in response to hormone stimulus, epithelial cell development, and host cell entry. Using bayes discriminant analysis, 48 signature genes were screened. We constructed a prognostic prediction system using the 48 signature genes and validated this system as relatively accurate and reliable. The DEGs might be closely associated with the prognosis in patients with breast cancer. We validated the effectiveness of our prognostic prediction system by GEO database. Therefore, this system might be a useful tool for preliminary screening and validation of potential prognosis indicators for ER+ breast cancer derived from mechanistic research.

Long Non-Coding RNA BANCR Is Overexpressed in Patients with Diabetic Retinopathy and Promotes Apoptosis of Retinal Pigment Epithelial Cells.

BACKGROUND lncRNA BANCR participates in the pathogenesis of various types of human diseases; however, its involvement in diabetic retinopathy is unknown. MATERIAL AND METHODS In this study, the expression of lncRNA BANCR in plasma of patients with diabetic retinopathy, diabetic patients without complications, and healthy controls was analyzed by qRT-PCR. The accuracy plasma BANCR in diagnosing diabetic retinopathy was analyzed by ROC curve analysis. lncRNA BANCR expression vector and siRNA were transfected into the human retinal pigment epithelial cell line ARPE-19, and cell apoptosis was analyzed by cell apoptosis assay. RESULTS We found that lncRNA BANCR was significantly upregulated in patients with diabetic retinopathy compared to diabetic patients without complications and healthy controls. Upregulation of lncRNA BANCR effectively distinguished patients with diabetic retinopathy from diabetic patients without complications and healthy controls. High-glucose treatment led to upregulated expression of BANCR in the human retinal pigment epithelial cell line ARPE-19. Overexpression of BANCR promoted and siRNA silencing inhibited the apoptosis of cells in the human retinal pigment epithelial cell line ARPE-19 in a high-glucose environment. CONCLUSIONS lncRNA BANCR is overexpressed in patients with diabetic retinopathy and can promote apoptosis of retinal pigment epithelial cells.

Identification of methylation sites and signature genes with prognostic value for luminal breast cancer.

BACKGROUND: Robust and precise molecular prognostic predictors for luminal breast cancer are required. This study aimed to identify key methylation sites in luminal breast cancer, as well as precise molecular tools for predicting prognosis. METHODS: We compared methylation levels of normal and luminal breast cancer samples from The Cancer Genome Atlas dataset. The relationships among differentially methylated sites, corresponding mRNA expression levels and prognosis were further analysed. Differentially expressed genes in normal and cancerous samples were analysed, followed by the identification of prognostic signature genes. Samples were divided into low- and high-risk groups based on the signature genes. Prognoses of low- and high-risk groups were compared. The Gene Expression Omnibus dataset were used to validate signature genes for prognosis prediction. Prognosis of low- and high-risk groups in Luminal A and Luminal B samples from the TCGA and the Metabric cohort dataset were analyzed. We also analysed the correlation between clinical features of low- and high- risk groups as well as their differences in gene expression. RESULTS: Fourteen methylation sites were considered to be related to luminal breast cancer prognosis because their methylation levels, mRNA expression and prognoses were closely related to each other. The methylation level of SOSTDC1 was used to divide samples into hypo- and hyper-methylation groups. We also identified an mRNA signature, comprising eight transcripts, ESCO2, PACSIN1, CDCA2, PIGR, PTN, RGMA, KLK4 and CENPA, which was used to divide samples into low- and high-risk groups. The low-risk group showed significantly better prognosis than the high-risk group. A correlation analysis revealed that the risk score was an independent prognostic factor. Low- and high- risk groups significantly correlated with the survival ratio in Luminal A samples, but not in Luminal B samples on the basis of the TCGA and the Metabric cohort dataset. Further functional annotation demonstrated that the differentially expressed genes were mainly involved in cell cycle and cancer progression. CONCLUSIONS: We identified several key methylation sites and an mRNA signature for predicting luminal breast cancer prognosis. The signature exhibited effective and precise prediction of prognosis and may serve as a prognostic and diagnostic marker for luminal breast cancer.

Clinical implications of pretreatment inflammatory biomarkers as independent prognostic indicators in prostate cancer.

OBJECTIVES: Research on the relationship between inflammatory biomarkers and malignant tumors has become a hotspot. Many studies have demonstrated that neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), and red blood cell distribution width (RDW) could act as independent prognostic indicators for several solid tumors. This study aimed to evaluate the clinical implications of pretreatment inflammatory biomarkers, including NLR, PLR, and RDW as independent prognostic indicators in prostate cancer (PCa). METHODS: A total of 226 PCa patients who were diagnosed at our institution from 2011 to 2016 were analyzed retrospectively. We compared the clinicopathological features, survival curves, and prognosis of the PCa patients between the high and low groups according to the cutoffs of NLR, PLR, and RDW. RESULTS: The pretreatment NLR, PLR, and RDW values were significantly higher in the patients with PCa than those in the controls (P<.05). Increased NLR and PLR values were significantly associated with high risk of progression, including higher Gleason scores, cell proliferation antigen 67 (Ki-67) indexes, and prostate-specific antigen (PSA) levels (P<.05), whereas an elevated RDW was only associated with an older age. An increased NLR was correlated with both overall survival (OS) (P=.025) and disease-free survival (DFS) (P=.017). In addition, a higher PLR only showed a significantly worse DFS (P=.040). Pretreatment NLR was an independent prognostic indicator of DFS. CONCLUSIONS: The pretreatment NLR and PLR might be beneficial to predict the progression and prognosis of PCa. Furthermore, NLR was more effective than PLR acting as an independent prognostic indicator for PCa.

Methyl eugenol induces potent anticancer effects in RB355 human retinoblastoma cells by inducing autophagy, cell cycle arrest and inhibition of PI3K/mTOR/Akt signalling pathway.

PURPOSE: Retinoblastoma is one of the lethal malignancies in children. Approximately half of the children that are diagnosed with retinoblastoma die of this disease. Enucleation is commonly used for the treatment of retinoblastoma but this leads to loss of vision. Therefore there is an urgent need to look for viable chemotherapeutic agents for the treatment of retinoblastoma. Consistent with this, natural products can produce efficient anticancer agents and in the present study a plant-derived phenylpropene methyl eugenol (ME) was evaluated against retinoblastoma RB355 cells. METHODS: The cytotoxic activity of this molecule was evaluated by MTT cell viability assay, while autophagic effects were evaluated by flow cytometry using acridine orange (AO)/monodansylcadaverine (AO/MDC) staining dyes. The effects on the cell cycle progression were analyzed by flow cytometry while the effects on mTOR/PI3K/Akt signalling pathway were assessed by western blot method. RESULTS: The results indicated that ME exhibited an IC50 value of 50 µM and exerted its cytotoxic effects in a dosedependent manner in RB355 cells. Moreover, it was observed that the ME lessens the cell viability and triggers G2/M cell cycle arrest. It was also observed that ME induced autophagy dose-dependently in retinoblastoma RB355 cells. Western blot analysis revealed that ME could modulate the mTOR/ PI3K/Akt signalling pathway in RB355 cells at the IC50 concentration. CONCLUSIONS: The above results clearly indicate that ME is a potent anticancer agent and may be developed further as a possible anticancer lead molecule against retinoblastoma provided further in depth in vivo studies are carried out.

Predictive value of serum pregnancy-associated plasma protein A for patients with ischemic cerebrovascular disease.

BACKGROUND: To investigate the role of pregnancy-associated plasma protein A (PAPP-A) in the outcome of ischemic cerebrovascular disease. METHODS: We analyzed the levels of PAPP-A in the transient ischemic attack (TIA) patients, ischemic stroke (IS) patients and normal control, and followed up the outcome of the patients in the following 2 years. Blood samples were drawn at admission, prior to treatment with heparins. RESULTS: The levels of PAPP-A in TIA patients, IS patients and normal control were 4.91 (2.11, 6.48) mIU/L, 6.77 (3.31, 10.23) mIU/L and 4.25 (1.76, 5.22) mIU/L, respectively. The follow-up results of TIA patients and IS patients indicated the PAPP-A concentration in the poor prognosis group were higher than those in the good prognosis group (5.90 vs 4.46 mIU/L, P<.05, 10.06 vs 5.12 mIU/L, P<.05, respectively). Serum PAPP-A concentration emerged as a predictor of risk stratification with an OR of 1.41 and 1.25 (P<.05, P<.05). CONCLUSIONS: Higher PAPP-A concentration has a forecasting value on prognosis in ischemic cerebrovascular disease.

Early occipital injury affects numerosity counting but not simple arithmetic.

This study investigated the effects of early occipital injury on the development of counting and simple arithmetic abilities in an occipital epileptic patient. This patient had obvious softening lesions in the bilateral occipital regions due to viral encephalitis at the age of 1.5 years. Results showed that she could perform subitizing and simple arithmetic very well, but could not perform numerosity counting tasks. These results suggest that the occipital cortex plays an important role in the development of numerosity counting skills, but not in the development of subitizing and simple arithmetic.