IL1R2 promotes retinal angiogenesis to participate in retinopathy of prematurity by activating the HIF1α/PFKFB3 pathway.

Retinopathy of prematurity (ROP) is the leading cause of blindness in children, but there is no safe and effective treatment available. Interleukin-1 receptor type 2 (IL1R2) acts as a decoy receptor for IL-1 may affect ROP progression. This study aimed to investigate the role of IL1R2 in ROP. A microglial cell model was established under hypoxia conditions and co-cultured with choroidal endothelial cells, while an oxygen-induced retinopathy (OIR) model was also established. Microglial activation and IL1R2 levels in retinal tissues were analyzed using immunofluorescence assay. Endothelial cell migration was evaluated by Transwell assay and scratch test, angiogenesis was assessed using ELISA and tube formation assay, and proliferation was evaluated by EdU assay. The HIF1α/PFKFB3 pathway was analyzed by western blot. We observed that IL1R2 expression was predicted to be upregulated in ROP and was increased in hypoxia-treated BV2 cells. Additionally, IL1R2 levels were upregulated in the retinal tissues of OIR mice and correlated with microglial activation. In vitro experiments, we found that hypoxia promoted endothelial cell migration, angiogenesis, proliferation, and activated the HIF1α/PFKFB3 pathway, which were rescued by IL1R2 knockdown. Moreover, NHWD-870 (a HIF1α/PFKFB3 pathway inhibitor) suppressed endothelial cell migration, angiogenesis, and proliferation induced by IL1R2 overexpression. In conclusion, IL1R2 facilitates the migration, angiogenesis, and proliferation of choroidal endothelial cells by activating the HIF1α/PFKFB3 pathway to regulate ROP progression.

Aromatic C-C Bond Cleavage in a Curved π-System of Aminocorannulene.

We report a metal- and oxidant-free aromatic C-C bond cleavage in the curved corannulene skeleton. Reaction of 1-aminocorannulene with hydrazonyl chloride generates an amidrazone intermediate that undergoes facile intramolecular proton migrations and ring annulation to give a 1,2,4-triazole derivative of planar benzo[ghi]fluoranthene, in which the release of strain associated with the curved π-surface and the formation of an aromatic triazole moiety are the driving forces. This report provides new insights into the aromatic C-C bond cleavage.

Therapeutic effects of allopurinol on the function of left ventricular and activity of matrix metalloproteinase enzymes (MMPs) in patients with chronic heart failure.

Heart failure is a growing public health problem, especially in the elderly, often occurring due to ischemia and coronary artery disease. Allopurinol can protect against myocardial ischemia and improve myocardial energy utilization during ischemia. On the other hand, matrix metalloproteinase (MMP) enzymes play an essential role in causing atherosclerosis, obstruction, and myocardial infarction. Therefore, in the present study, the effect of allopurinol on the function of the left ventricular and the activity of MMP-1, MMP-2, MMP-3, and MMP-9 were evaluated in heart failure patients. In this clinical trial, 82 patients were randomly assigned to allopurinol or placebo in addition to standard treatment. Echocardiographic evaluations were performed before treatment and six months after treatment. Also, after allopurinol treatment, plasma and peripheral blood mononuclear cells were extracted from control and intervention groups. The active form of MMPs was measured by ELISA and mRNA expression by Real-time PCR. The rate of change in left ventricular ejection fraction in the allopurinol group was significantly higher than patients in the control group. There was also found more improvement in NYHA class in patients receiving allopurinol than in the control group. ELISA results showed that all plasma MMP levels in the control group were significantly higher than those in the allopurinol group (P<0.001). Quantitative determination of mRNA expression in MMPs by Real-time RT-PCR revealed that, except for MMP-9, there was no significant difference in the expression of evaluated MMPs between the treatment and control groups. In general, the results showed that long-term administration of allopurinol improves left ventricular function, and it has beneficial effects on the life quality of patients with heart failure.

Clinical implication of aquaporin 9 in non-small cell lung cancer patients: its expression and relationship with clinical features and prognosis.

BACKGROUND: Aquaporin 9 (AQP9) is recognized as a key regulator in several cancers, whereas little is known about its clinical implication in non-small cell lung cancer (NSCLC). Thus, we aimed to explore AQP9 expression and its relationship with clinical features, prognosis in NSCLC patients. METHODS: One hundred ninety-eight NSCLC patients who received resection were retrospectively enrolled. This study contained two cohorts: in cohort A, AQP9 protein expression (from formalin fixed paraffin embedded tumor and paired adjacent tissue specimens) in 198 patients was detected by immunohistochemistry (IHC). In cohort B, AQP9 mRNA expression (from fresh-frozen tumor and paired adjacent tissues) in 108 patients (out of 198 patients) was detected by RT-qPCR. RESULTS: In cohort A, increased AQP9 IHC score and greater proportion of AQP9 protein high expression cases were shown in tumor tissue than adjacent tissue (both P < 0.001). Tumor AQP9 protein high expression correlated with lymph node (LYN) metastasis (P = 0.002) and raised TNM stage (P = 0.012). Interestingly, tumor AQP9 protein high expression related to worse disease-free survival (DFS) (P = 0.002) and overall survival (OS) (P = 0.026). In cohort B, AQP9 mRNA expression in tumor tissue was increased than adjacent tissue (P < 0.001), and tumor AQP9 mRNA high expression linked to LYN metastasis (P = 0.024) and increased TNM stage (P = 0.032) as well; tumor AQP9 mRNA high expression was related to shorter DFS (P = 0.009), and it presented with a trend to be correlated with worse OS (P = 0.054), but without statistical significance. CONCLUSION: AQP9 serves as a potential indicator for monitoring disease progression and prognostication in NSCLC patients.

MiR-181b serves as diagnosis and prognosis biomarker in severe community-acquired pneumonia.

Severe community-acquired pneumonia (SCAP) is a common critical disease in the intensive care unit (ICU). This study aims to evaluate the clinical significance of miR-181b in SCAP, which has been revealed to be dysregulated in acute respiratory distress syndrome events due to SCAP. There were 50 SCAP patients and 26 healthy volunteers were recruited in this study. The expression of miR-181b was detected by RT-qPCR and the difference between SCAP and healthy controls was evaluated. The diagnosis and prognosis value of miR-181b was assessed by the receiver operating characteristics (ROC), Kaplan-Meier, and Cox regression analysis. miR-181b was significantly downregulated in SCAP compared with healthy controls. The downregulation of miR-181b showed a significant association with the white blood cell count, absolute neutrophils, and the C-reactive protein of patients. The downregulation of miR-181b could distinguish SCAP patients from healthy controls and predicate the poor prognosis of SCAP patients. Downregulated miR-181b serves as a diagnosis and prognosis biomarker for SCAP, which may be useful biological information for the early detection and risk estimation of SCAP.

KMT2D promotes proliferation of gastric cancer cells: evidence from ctDNA sequencing.

BACKGROUND: ctDNA sequencing could be used for early cancer screening, prognosis prediction, and medication guidance. However, data of its application in gastric cancer are still lacking. In this study, using ctDNA sequencing, we aimed to screen the mutant genes closely associated with gastric cancer and to explore the impact of these genes on gastric cancer development. METHODS: ctDNA for high-throughput sequencing was obtained from gastric cancer patients, and the high-frequency mutant gene KMT2D was identified. Immunohistochemical examination was conducted to assess the expression of KMT2D in gastric cancer tissues. KMT2D knockdown was performed to establish the stably transfected gastric cancer cells. Then, CCK8, plate clone formation assay, and Transwell assay were conducted, and a subcutaneous tumor-bearing model was induced in nude mice to investigate the changes in cell proliferation and invasion capability. Transcriptome sequencing was also performed to investigate the differences in cellular gene expression. RESULTS: Detection of ctDNA found 113 gastric cancer-related mutations, 11 of which are the top 20 high-frequency mutations of gastric cancer recorded by COSMIC (Catalogue of Somatic Mutations in Cancer, COSMIC). They are TP53, ARID1A, CDH1, PIK3CA, KMT2C, KMT2D, APC, SPEN, CTNNB1, SETBP1, and KMT2A. The gene closely related to the clinical characteristics of the patient is KMT2D. The high-frequency mutant gene KMT2D was identified in gastric cancer tissues. The positive rate of KMT2D expression in cancer tissues was 74.3%, which was higher than that in para-carcinoma tissues (56.8%). The knockdown of KMT2D inhibited the proliferation, invasion, and tumor formation capacity of the gastric cancer cells, causing differences in the gene expression profiles, and the expression of different functional gene clusters was up- or downregulated. CONCLUSION: The findings of this study revealed that KMT2D could be an oncogene capable of promoting gastric cancer proliferation.

Down-Regulation of TMPO-AS1 Induces Apoptosis in Lung Carcinoma Cells by Regulating miR-143-3p/CDK1 Axis.

Evidence has shown that long non-coding RNAs (lncRNA) play pivotal roles in cancer promotion as well as suppression. But the molecular mechanism of lncRNA TMPO antisense transcript 1 (TMPO-AS1) in lung cancer (LC) remains unclear. This study mainly investigated the effect of TMPO-AS1 in LC treatment. TMPO-AS1 was tested by Kaplan-Meier method. Quantitative real time polymerase chain reaction (qRT-PCR) was employed to assess the expressions of TMPO-AS1, miR-143-3p, and CDK1 respectively in LC tissues and cell lines. TMPO-AS1, miR-143-3p and CDK1 expressions in LC cells were regulated through cell transfection, followed by MTT for cell viability detection. Besides, dual-luciferase reporter assays were performed to verify the interrelated microRNA of TMPO-AS1 and the target of miR-143-3p. Western blot experiments were used to examine the expressions of apoptosis-related proteins, and flow cytometry tested the cell apoptosis in treated cells. TMPO-AS1 and CDK1 were overexpressed in LC tissues and cells, while miR-143-3p level was suppressed. The decrease of TMPO-AS1 led to the increase of miR-143-3p, which further resulted in the reduction of CDK1. Down-regulating TMPO-AS1 reduced LC cell viability, motivated cell apoptosis, as well as promoted the expressions of Bcl and CCND1 and restrained Caspase-3 level, but all these consequences were abrogated by miR-143-3p inhibitor. Simultaneously, siCDK1 could reverse the anti-apoptosis and pro-activity functions of miR-143-3p inhibitor in LC cells. Down-regulation of TMPO-AS1 has the effects of pro-apoptosis in LC by manipulating miR-143-3p/CDK1, which is hopeful to be a novel therapy for LC patients.

Silencing of BRF2 inhibits the growth and metastasis of lung cancer cells.

Transcription factor II B (TFIIB)­related factor 2 (BRF2) is involved in the development of cancer, but its role in lung cancer is underreported. The present study aimed to explore the role of BRF2 in the regulation of lung cancer cells. Immunofluorescence staining and immunohistochemistry were performed to detect BRF2 protein expression in human lung cancer cells and tissues. Following cell transfection with small interfering RNA for silencing BRF2, the cell proliferation was examined by Cell Counting Kit­8 and MTT assays. Cell apoptosis, migration and invasion were determined by flow cytometry, wound­healing and Transwell assay. The expression levels of Akt, phosphorylated (p)­Akt, Bax, E­cadherin, Bcl­2, N­cadherin, Snail and epidermal growth factor receptor (EGFR) in human lung cancer A549 cells were detected by western blotting. The results demonstrated that BRF2 expression was increased in human lung cancer cells and tissues, and that silencing of BRF2 promoted cell apoptosis but inhibited cell proliferation and migration. The protein expression levels of Akt, E­cadherin, p­Akt, Bcl­2, N­cadherin, Snail and EGFR in A549 cells were inhibited by silencing of BRF2, while expression levels of Bax and E­cadherin were increased by silencing BRF2. In conclusion, BRF2 demonstrates high expression in lung cancer and silencing of BRF2 inhibits the growth and metastasis of lung cancer cells. The current findings provide a novel approach for the treatment of lung cancer.

Comprehensive Analysis of CDC27 Related to Peritoneal Metastasis by Whole Exome Sequencing in Gastric Cancer.

INTRODUCTION: The peritoneum is the most common metastatic site of gastric cancer and is associated with a dismal prognosis. However, there is no reliable biomarker for predicting peritoneal metastasis (PM). MATERIALS AND METHODS: Whole-exome sequencing (WES) was performed on formalin-fixed, paraffin-embedded (FFPE) samples from 63 patients with stage I-III gastric cancer and circulating tumor DNA (ctDNA) samples from 10 patients with stage IV gastric cancer. Differentially expressed genes (DEGs) were identified between the PM and non-PM groups and analyzed by multiple bioinformatics analyses. Univariate and multivariate Cox regression analyses were used to identify the risk factors for PM and a risk score model was developed. RESULTS: The number of mutant genes and the tumor mutation burden (TMB) in the PM group were higher than those in the non-PM group (p < 0.05). There was a significant positive correlation between the number of mutant genes and the TMB (R2 = 0.9997). The risk of PM was significantly higher in the high TMB group than in the low TMB group (p = 0.045). Forty-nine DEGs were identified as associated with PM in gastric cancer. CDC27 mutations were associated with a higher risk for PM and poor survival. The CDC27 mutations were located in the Apc3 region, the TPR region, and the phosphorylation region, and new mutation sites were not included in the TCGA database. Multivariable Cox regression analysis demonstrated that pathological T stage, poor tumor differentiation, Borrmann type, and CDC27 mutations were independent predictive factors of PM. A risk score model was constructed that demonstrated good performance. CONCLUSION: Through WES, we identified 49 DEGs relevant to PM in gastric cancer. CDC27 mutations were independently associated with PM by statistical and bioinformatics analyses. A risk score model was built and was demonstrated to effectively discriminate gastric cancer patients with and without PM.

Detection of circulating tumor cell DNA for monitoring advanced gastric cancer.

INTRODUCTION: Circulating tumor DNA (ctDNA) for monitoring the effects of chemotherapy and predicting prognosis in advanced gastric cancer have not been thoroughly investigated. METHODS: We performed next-generation sequencing (NGS) of ctDNA from 23 gastric cancer patients. Then the genetic information and clinical information were statistically analyzed. RESULTS: In this study, the frequency of TP53 was significantly different between the effective and ineffective groups (P = 0.040), and the number of TP53 mutations was more frequent in the ineffective group. Missense mutation was a significant difference between the treatment effect groups (P = 0.026). The number of gene mutations and the change in copy number levels were related to therapeutic effect. Among the ineffective group, there was a significant difference in the number of gene mutations (P = 0.0006). We further divided the number of gene mutations into an increase group and a decrease group, and found that there was a significant difference between the effective and ineffective groups (P = 0.038). Finally, it was found that patients with high mutation abundance of gastric cancer had a shorter overall survival than patients with low mutation abundance (P<0.05). CONCLUSION: ctDNA can be used as an effective tool to monitor the efficacy of chemotherapy and predict prognosis in advanced gastric cancer.

Molecular Profiles and Metastasis Markers in Chinese Patients with Gastric Carcinoma.

The goal of this work was to investigate the molecular profiles and metastasis markers in Chinese patients with gastric carcinoma (GC). In total, we performed whole exome sequencing (WES) on 74 GC patients with tumor and adjacent normal formalin-fixed, paraffin-embedded (FFPE) tissue samples. The mutation spectrum of these samples showed a high concordance with TCGA and other studies on GC. PTPRT is significantly associated with metastasis of GC, suggesting its predictive role in metastasis of GC. Patients carrying BRCA2 mutations tend not to metastasize, which may be related to their sensitivity to chemotherapy. Mutations in MACF1, CDC27, HMCN1, CDH1 and PDZD2 were moderately enriched in peritoneal metastasis (PM) samples. Furthermore, we found two genomic regions (1p36.21 and Xq26.3) were associated with PM of GC, and patients with amplification of 1p36.21 and Xq26.3 have a worse prognosis (P = 0.002, 0.01, respectively). Our analysis provides GC patients with potential markers for single and combination therapies.

Publisher Correction: A multistage rotational speed changing molecular rotor regulated by pH and metal cations.

The original version of this Article omitted the Received and Accepted dates; they should have been 22nd January 2018 and 19th April 2018, respectively. This has been corrected in the PDF and HTML versions of the Article.

A multistage rotational speed changing molecular rotor regulated by pH and metal cations.

Despite having significant applications in building nanomachines, molecular rotors with the rotational speed modulations to multiple stages in a wide range of frequency have not yet been well established. Here, we report the discovery of a stimuli-responsive molecular rotor, the rotational speed of which in the slow-to-fast range could be modulated to at least four stages triggered by acid/base and metal cations. The rotor itself rotates rapidly at ambient or elevated temperature but displays a restricted rotation after deprotonation due to the produced intramolecular electrostatic repulsion. Subsequent addition of Li+ or Na+ cations introduces an electrostatic bridge to stabilize the transition state of the deprotonated rotor, thus giving a cation-radius-dependent acceleration of the rotation to render the rotor running at a mid-speed. All the stimuli are highly reversible. Our studies provide a conceptual approach for constructing multistage rotational-speed-changing molecular rotors, and further, the practical nanomachines.

Pulmonary alveolar proteinosis with peripheral adenocarcinom.

Background: Pulmonary alveolar proteinosis (PAP) is a rare interstitial lung disease classified into congenital form, autoimmune form and secondary form. Secondary PAP is caused by underlying conditions including solid malignancies. Few cases reported PAP associated with lung cancers. Objective: To show the clinical features of PAP with adenocarcinom, tried to seek for possible mechanism to explain whole clinical course. Methods: Reported a case of PAP associated with lung adenocarcinom, and also reviewed the relevant literature on PAP. Results: The patient suffered from intermittent cough, fever, shortness of breath, thoracalgia or hemoptysis. Blood gas analysis showed hyoxemia. Spirometric abnormality is mildly restrictive defect. High-resolution computed tomography (HRCT) showed patchy, ground-glass opacities with interlobular septal thickening called as "crazy-paving" pattern. Positron emission tomography/computed tomography (PET/CT) revealed a nodule with characteristics of lobulation and spiculation in the right lung apex section and diffuse consolidation shadow spreading over rest of lung field. Bronchoalveolar lavage fluid (BALF) showed a large amount of amorphous red-dyed materials and a few alveolar macrophages scattered in endoalveolar space with PAS positive. Transbronchial lung biopsy found adenocarcinoma. Wedge resection with mediastinal lymphnode and then 2 cycles of postoperative chemotherapy was carried out. No ground-glass opacities were found in his chest CT pictures in the next nine months. This result may support the theory that lung cancer cells cause quantitative or functional damage to alveolar macrophages, which trend to secondary PAP. Conclusions: The patient had typical clinical features of PAP. PAP may be secondary to lung cancer. (Sarcoidosis Vasc Diffuse Lung Dis 2018; 35: 390-394).

Aromatic triazole foldamers induced by C-H···X (X = F, Cl) intramolecular hydrogen bonding.

Aryl-triazole oligomers based on isobutyl 4-fluorobenzoate and isobutyl 4-chlorobenzoate were designed and synthesized. Crystal structure and (1)H-(1)H NOESY experiments demonstrate that the oligomers adopt stable helical conformation, which are induced by C(5)-H···X-C (X = F, Cl) intramolecular hydrogen bonding between triazole protons and halogen atoms. The stabilities of the folded conformations are confirmed by DFT calculations, which show that each C(5)-H···F-C planar interaction lowers the energy by ~3 kcal mol(-1) on average, and by ~1 kcal mol(-1) when C(5)-H···Cl-C bridges are formed. The hydrogen-bonding networks are disrupted in competitive hydrogen-bonding media such as DMSO, generating the unfolded oligomers.

[A retrospective study on the progression of myopia in school-age children].

OBJECTIVE: To study the progression of myopia in school-age children over the past 12 years and factors influencing myopia progression. METHODS: A total of 4569 cases of 5 to 12-year-old children who had refractive examinations in the Third Xiangya Hospital, Central South University between January 2000 and December 2011 were enrolled in this study. The children had no family history of congenital high myopia or other eye diseases. Myopia progression was evaluated when the children were re-examined. The refractive state of each child was measured with cyclopiegic retinoscopy. RESULTS: The mean spherical equivalent (SE) myopia was-2.0±1.7 D between January 2000 and December 2011. There was no statistical difference in yearly myopia progression between different years. The average age of the myopic children decreased from 10.1 in 2000 to 8.9 years old in 2011 (P<0.05). Mean myopia progression was -0.6±0.7 D per year from 2000 to 2011. Myopia progression reduced gradually in 5 to 8-year-olds (P<0.05), however, it accelerated between ages 9 and 11 years. Myopia progression in 10- and 11-year-olds was significantly greater than in 7- and 8-year-olds (P<0.01). The multiple linear regression analysis demonstrated that age and baseline myopic refraction were positively related to myopia progression. CONCLUSIONS: There was no obvious change in the yearly myopia progression of the children over the past 12 years. The mean age of myopia occurrence became younger with time. More preventive measures are needed to ward off high myopia in children with moderate myopia, especially those aged over 10 years.

[Significance of the expression of lymphocytes and cytokines infiltrating in HCC].

AIM: To reveal the classification of lymphocytes and expression of cytokines infiltrating in HCC, and investigate the significance of changes of immune microenvironment in HCC. METHODS: 76 tumor and non-tumor tissues of HCC were collected to detect the amount of immune cells in the tissues by FCM and immunohistochemistry (IHC). Th1/Th2 cytokines in tissues were detected by Cytometric bead array (CBA), and TGF-1, VEGF detected by ELISA. RESULTS: There were more CD3(+); T cells, CD4(+); Th cells, Treg cells, and CD45RO(+); memory T cells in tumors than in non-tumor tissues. On the contrary there were less CD8(+); CTLs in tumors. There was negative correlation between Treg and Th or CTL cells. CD69, which is the early activating factor, expressed less on CD3(+); T cells and NK cells in tumors than non-tumor tissues. Whereas HLA-DR, which is the late activating factor, expressed more on CD3(+); T cells in tumors than non-tumor tissues. More IL-10, TGF-1 and VEGF were secreted in tumors than non-tumor tissues. CONCLUSION: Along with the decrease of effective immune cells and increase of suppressor immune cells and cytokines, the lymphocytes infiltrating in the tumor were immune incompitence, which contributed to the tumorigenesis.

Deoxyuridines from the marine sponge associated actinomycete Streptomyces microflavus.

One new nucleoside derivative, named 3-acetyl-5-methyl-2'-deoxyuridine (1), along with two known compounds 3,5-dimethyl-2'-deoxyuridine (2) and 3-methyl-2'-deoxyuridine (3), were isolated from the cultures of Streptomyces microflavus. This strain was an associated actinomycete isolated from the marine sponge Hymeniacidon perlevis collected from the coast of Dalian (China). Their structures were elucidated by detailed NMR and MS spectroscopic analysis as well as comparison with literature data.