Antenatal corticosteroid therapy in late preterm delivery: a nationwide population-based retrospective study in Taiwan.

OBJECTIVE: To investigate whether antenatal corticosteroid therapy improves neonatal and maternal outcomes in late preterm delivery. DESIGN: Population-based retrospective study. SETTING: The linkages of Taiwan's National Health Insurance Research Database, National Birth Reporting Database, and the Taiwan Maternal and Child Health Database. POPULATION: All births at risk for late preterm deliveries in Taiwan between 2004 and 2011. METHODS: For every birth at risk for late preterm delivery, five controls randomly matched by maternal and gestational ages and birthweight were included. A conditional logistic regression analysis was applied for risk estimation, with births without corticosteroids as the reference group. Odds ratios were adjusted for caesarean section, parity, sex, gestational hypertension and gestational diabetes mellitus. MAIN OUTCOME MEASURES: Neonatal outcomes, maternal outcomes and the utilisation of healthcare services. RESULTS: The outcomes of 5745 women treated with corticosteroids between 34+0  weeks and 36+6  weeks of gestation were compared with those of 28 135 untreated controls. Compared with the controls, births from women administered corticosteroids reduced the need for continuous positive airway pressure, the number of neonatal intensive care unit admission, and the need for glucose administration, as well as the risk of neonatal respiratory distress, but increased the risk of neonatal sepsis and the number of outpatient visits. CONCLUSIONS: Antenatal corticosteroid therapy in women at risk of late preterm delivery may significantly reduce the need for respiratory support and glucose supply, and respiratory complication risk in neonates. TWEETABLE ABSTRACT: Antenatal corticosteroids in late preterm delivery reduced the risk of neonatal respiratory complications in Taiwan.

A roadmap to build a phenotypic metric of ageing: insights from the Baltimore Longitudinal Study of Aging.

Over the past three decades, considerable effort has been dedicated to quantifying the pace of ageing yet identifying the most essential metrics of ageing remains challenging due to lack of comprehensive measurements and heterogeneity of the ageing processes. Most of the previously proposed metrics of ageing have been emerged from cross-sectional associations with chronological age and predictive accuracy of mortality, thus lacking a conceptual model of functional or phenotypic domains. Further, such models may be biased by selective attrition and are unable to address underlying biological constructs contributing to functional markers of age-related decline. Using longitudinal data from the Baltimore Longitudinal Study of Aging (BLSA), we propose a conceptual framework to identify metrics of ageing that may capture the hierarchical and temporal relationships between functional ageing, phenotypic ageing and biological ageing based on four hypothesized domains: body composition, energy regulation, homeostatic mechanisms and neurodegeneration/neuroplasticity. We explored the longitudinal trajectories of key variables within these phenotypes using linear mixed-effects models and more than 10 years of data. Understanding the longitudinal trajectories across these domains in the BLSA provides a reference for researchers, informs future refinement of the phenotypic ageing framework and establishes a solid foundation for future models of biological ageing.

Hypoxic lung cancer-secreted exosomal miR-23a increased angiogenesis and vascular permeability by targeting prolyl hydroxylase and tight junction protein ZO-1.

Hypoxia plays a critical role during the evolution of malignant cells and tumour microenvironment (TME).Tumour-derived exosomes contain informative microRNAs involved in the interaction of cancer and stromal cells, thus contributing to tissue remodelling of tumour microenvironment. This study aims to clarify how hypoxia affects tumour angiogenesis through exosomes shed from lung cancer cells. Lung cancer cells produce more exosomes under hypoxic conditions than do parental cells under normoxic conditions. miR-23a was significantly upregulated in exosomes from lung cancer under hypoxic conditions. Exosomal miR-23a directly suppressed its target prolyl hydroxylase 1 and 2 (PHD1 and 2), leading to the accumulation of hypoxia-inducible factor-1 α (HIF-1 α) in endothelial cells. Consequently, hypoxic lung cancer cells enhanced angiogenesis by exosomes derived from hypoxic cancer under both normoxic and hypoxic conditions. In addition, exosomal miR-23a also inhibits tight junction protein ZO-1, thereby increasing vascular permeability and cancer transendothelial migration. Inhibition of miR-23a by inhibitor administration decreased angiogenesis and tumour growth in a mouse model. Furthermore, elevated levels of circulating miR-23a are found in the sera of lung cancer patients, and miR-23a levels are positively correlated with proangiogenic activities. Taken together, our study reveals the clinical relevance and prognostic value of cancer-derived exosomal miR-23a under hypoxic conditions, and investigates a unique intercellular communication, mediated by cancer-derived exosomes, which modulates tumour vasculature.

Elevated hydrostatic pressure enhances the motility and enlarges the size of the lung cancer cells through aquaporin upregulation mediated by caveolin-1 and ERK1/2 signaling.

The mechanical characteristics presented in cancer microenvironment are known to have pivotal roles in cancer metastasis, which accounts for the leading cause of death from malignant tumors. However, while a uniformly distributed high interstitial fluid pressure (IFP) is a common feature in solid tumors, the effects of high IFP on the motility and invasiveness of cancer cells remain obscure. Using cell-culture devices that simulated increased IFP conditions by applying hydrostatic pressure (HP) ranging from 0 to 20 mm Hg to the cells, we found that the elevated HPs increased the migration speeds, invasiveness, cell volume, filopodial number and aquaporin-1 (AQP1), Snail and vinculin expression levels, as well as phosphorylation of caveolin-1 and extracellular signal-regulated kinase1/2 (ERK1/2), in the lung cancer cells CL1-5 and A549. The increases of migration speed and cell volume correlated temporally with the increase of AQP1 expression. The elevated HP-induced migration acceleration was hindered by AQP1 knockdown using small interfering RNA (siRNA) transfection. Inhibition of ERK1/2 phosphorylation using the mitogen-activated protein kinase kinase inhibitor PD98059 abrogated the elevated HP-induced AQP1 upregulation and migration acceleration in the cancer cells. Caveolin-1 knockdown by siRNA transfection attenuated the HP-induced, ERK1/2-depedent AQP1 upregulation and migration acceleration. Further biochemical studies revealed that the caveolin-1 activation-driven ERK1/2 signaling is mediated by Akt1/2 phosphorylation. By contrast, the elevated HPs had negligible effects on the migration speed and volume of normal bronchial epithelial cells. These results disclose a novel mechanism relating high IFP to the invasiveness of cancer cells and highlight potential targets to impede cancer spreading.

Angiomotin decreases lung cancer progression by sequestering oncogenic YAP/TAZ and decreasing Cyr61 expression.

Lung cancer is the leading cause of cancer death worldwide, with metastasis underlying majority of related deaths. Angiomotin (AMOT), a scaffold protein, has been shown to interact with oncogenic Yes-associated protein/transcriptional co-activator with a PDZ-binding motif (YAP/TAZ) proteins, suggesting a potential role in tumor progression. However, the functional role of AMOT in lung cancer remains unknown. This study aimed to identify the patho-physiological characteristics of AMOT in lung cancer progression. Results revealed that AMOT expression was significantly decreased in clinical lung cancer specimens. Knockdown of AMOT in a low metastatic CL1-0 lung cancer cell line initiated cancer proliferation, migration, invasion and epithelial-mesenchymal transition. The trigger of cancer progression caused by AMOT loss was transduced by decreased cytoplasmic sequestration and increased nuclear translocation of oncogenic co-activators YAP/TAZ, leading to increased expression of the growth factor, Cyr61. Tumor promotion by AMOT knockdown was reversed when YAP/TAZ or Cyr61 was absent. Further, AMOT knockdown increased the growth and spread of Lewis lung carcinoma in vivo. These findings suggest that AMOT is a crucial suppressor of lung cancer metastasis and highlight its critical role as a tumor suppressor and its potential as a prognostic biomarker and therapeutic target for lung cancer.

Direct duplication of the Y chromosome with normal phenotype - incidental finding in two cases.

Structural rearrangement in the Y chromosome is closely involved in spermatogenesis. However, several Y chromosome variants may have no deleterious effects on male reproduction. Here, we report two cases of Y chromosomal duplication from incidental findings. Their FISH analysis revealed direct duplication of large segments of short and long arms of the Y chromosome. Nearly two intact Y chromosomes were carried in these two cases with normal phenotype.

Breast tumor-associated osteoblast-derived CXCL5 increases cancer progression by ERK/MSK1/Elk-1/snail signaling pathway.

The skeleton is the most common metastatic site for breast cancer, with bone metastasis causing pain as well as risk of pathological fractures. Interaction between tumors and the bone microenvironment creates a vicious cycle that accelerates both bone destruction and cancer progression. This study is the first to analyze the soluble factors secreted by breast tumor-associated osteoblasts (TAOBs), which are responsible for promoting cancer progression. The addition of CXCL5 (chemokine (C-X-C motif) ligand 5), present in large amounts in TAOB-condition medium (TAOB-CM), mimicked the inductive effect of TAOB-CM on breast cancer epithelial-mesenchymal transition, migration and invasion. In contrast, inhibition of CXCL5 in OBs decreased TAOB-mediated cancer progression. Inducement of MCF-7 and MDA-MB-231 cancer progression by TAOB-derived CXCL5 is associated with increased Raf/MEK/ERK activation, and mitogen- and stress-activated protein kinase 1 (MSK1) and Elk-1 phosphorylation, as well as Snail upregulation. Activation of Elk-1 facilitates recruitment of phosphorylated MSK1, which in turn enhances histone H3 acetylation and phosphorylation (serine 10) of Snail promoter, resulting in Snail enhancement and E-cadherin downregulation. Moreover, mice treated with anti-CXCL5 antibodies showed decreased metastasis of 4T1 breast cancer cells. Our study suggests that inhibition of CXCL5-mediated ERK/Snail signaling is an attractive therapeutic target for treating metastases in breast cancer patients.

Signalling pathway of isophorone diisocyanate-responsive interleukin-8 in airway smooth muscle cells.

This study is the first to analyse the soluble factors secreted by the bronchial epithelium after exposure to isophorone diisocyanate (IPDI) that are responsible for increasing migration and proliferation of primary normal human bronchial smooth muscle cells (BSMCs). We treated immortalised, nontumorigenic human bronchial epithelial cells (cell line BEAS-2B) and primary normal human bronchial epithelial cells (HBEC) with IPDI, and then collected the conditioned culture media (IPDI-BEAS-2B-CM and IPDI-HBEC-CM, respectively), which was added to BSMCs. Exposure of BEAS-2B cells and HBECs to IPDI increased interleukin (IL)-8 production. Culture of BSMCs with IPDI-BEAS-2B-CM and IPDI-HBEC-CM increased BSMC proliferation and migration, which are major features in asthma-related airway remodelling. Induction of BSMC proliferation and migration by IPDI-BEAS-2B-CM and IPDI-HBEC-CM was associated with increased focal adhesion kinase (FAK), Src, extracellular signal-regulated kinase (ERK)1/2 and AKT activation. Blocking FAK with a specific inhibitor significantly decreased BSMC migration and proliferation by inhibiting ERK1/2 activation. FAK and ERK1/2 inhibitor also decreased IPDI-BEAS-2B-CM-, IPDI-HBEC-CM- and recombinant human IL-8-mediated BSMC proliferation and migration, whereas blocking Rnd3 using small interfering RNA failed to affect BSMC proliferation, suggesting that Rnd3 was only involved in the regulation of BSMC migration. Our study suggests that inhibition of IL-8 or IL-8-mediated FAK/ERK/Rnd3 signalling is an attractive therapeutic target for IPDI-mediated asthma.

Identification and characterization of a novel Rab GTPase-activating protein in spermatids.

We have previously identified novel testis-specific genes by microarray analysis of human testicular tissues. One of the novel genes is Male Germ Cells Rab GTPase- Activating Proteins (MgcRabGAP), which is characterized by the conserved RabGAP catalytic domain, TBC (Tre2/Bub2/Cdc16). RabGAPs are involved in various physiological processes (e.g. vesicular trafficking, cytoskeletal remodelling, cell migration, etc.) by inactivating Rab proteins. In this study, we found that MgcRabGAP transcripts are mainly expressed in the mouse and human testes. The MgcRabGAP protein is expressed in the elongating and elongated spermatids. Immunofluorescence assay of mouse germ cells showed that the protein expression is enriched at the edge of the acrosomal region, neck and annulus during spermiogenesis. This MgcRabGAP is co-localized with its candidate substrate Rab3A at the acrosome/acroplaxome and neck regions of spermatids. Meanwhile, MgcRabGAP is co-localized and interacts with ß-actin. In humans, the expression of MgcRabGAP is enriched at the stage of elongating spermatids. The amount of MGCRABGAP transcript is reduced in the testicular tissues of men with various types of spermatogenic defects. Considering that MGCRABGAP is exclusively expressed in post-meiotic male germ cells, the decreased transcript amount may be a phenomenon secondary to loss of germ cells in the testicular samples. Our finding strongly suggests that MgcRabGAP is involved in acrosome/acroplaxome formation and cytoskeletal reorganization via Rab activity during mammalian spermiogenesis.

Chalcone inhibits the proliferation of human breast cancer cell by blocking cell cycle progression and inducing apoptosis.

Chalcones are discussed to represent cancer preventive food components in a human diet that is rich in fruits and vegetables. In this study, we examined chalcone (1,3-diphenyl-2-propenone) for its effect on proliferation in human breast cancer cell lines, MCF-7 and MDA-MB-231. The results showed that chalcone inhibited the proliferation of MCF-7 and MDA-MB-231 by inducing apoptosis and blocking cell cycle progression in the G2/M phase. Immunoblot assay showed that chalcone significantly decreased the expression of cyclin B1, cyclin A and Cdc2 protein, as well as increased the expression of p21 and p27 in a p53-independent manner, contributing to cell cycle arrest. An enhancement in Fas/APO-1 and its two form ligands, membrane-bound Fas ligand (mFasL) and soluble Fas ligand (sFasL), was responsible for the apoptotic effect induced by chalcone. In addition, chalcone also triggered the mitochondrial apoptotic signaling by increasing the amount of Bax and Bak and reducing the level of Bcl-2 and Bcl-X(L), and subsequently activated caspase-9 in MCF-7 and MDA-MB-231 cells. Taken together, our study suggests that the blockade of cell cycle progression and initiation of cell apoptotic system may participate in the antiproliferative activity of chalcone in human breast cancer cells.

The antiproliferative activity of prodelphinidin B-2 3'-O-gallate from green tea leaf is through cell cycle arrest and Fas-mediated apoptotic pathway in A549 cells.

Prodelphinidin B-2 3'-O-gallate, a proanthocyanidin gallate isolated from green tea leaf, was investigated for its anti-proliferative activity in human non-small cell lung cancer A549 cells. The results showed that prodelphinidin B-2 3'-O-gallate inhibited the proliferation of A549 cells with no detectable toxic effects on normal WI-38 cells as measured by the XTT assay. Flow cytometric analysis showed that prodelphinidin B-2 3'-O-gallate blocked cell cycle progression in the G0/G1 phase. In addition, prodelphinidin B-2 3'-O-gallate effectively induced A549 cell apoptosis as determined by assessing the nucleosome level in cytoplasm. Enzyme-linked immunosorbent assay showed that the G0/G1 phase arrest is due to p53-independent induction of p21/WAF1. An enhancement in Fas/APO-1 and its two form ligands, membrane-bound Fas ligand (mFasL) and soluble Fas ligand (sFasL), might be responsible for the apoptotic effect induced by prodelphinidin B-2 3'-O-gallate. We suggested that prodelphinidin B-2 3'-O-gallate's activities might be potentially contribute to its overall chemopreventive effects against lung cancer, and can possibly be considered for future therapeutic application.

Green tea constituent (-)-epigallocatechin-3-gallate inhibits Hep G2 cell proliferation and induces apoptosis through p53-dependent and Fas-mediated pathways.

(-)-Epigallocatechin-3-gallate (EGCG) is a polyphenolic compound found in green tea. It has been reported to possess a wide range of pharmacological properties, and is one of the most promising chemopreventive agents for cancer. To provide a better understanding of the preventive effect of EGCG on liver cancer, we examined EGCG for its effect on proliferation and cell cycle progression in a human liver cancer cell line, Hep G2. The results showed that EGCG inhibited the proliferation of Hep G2 by inducing apoptosis and blocking cell cycle progression in the G1 phase. ELISA showed that EGCG significantly increased the expression of p53 and p21/WAF1 protein, and this contributed to cell cycle arrest. An enhancement in Fas/APO-1 and its two form ligands, membrane-bound Fas ligand (mFasL) and soluble Fas ligand (sFasL), as well as Bax protein, was responsible for the apoptotic effect induced by EGCG. Taken together, our study suggests that the induction of p53 and the activity of the Fas/FasL apoptotic system play major roles in the antiproliferative activity of EGCG in Hep G2 cells.

Molecular characterization of deletional forms of beta-thalassemia in Taiwan.

beta-Thalassemia is one of the most common genetic diseases in Taiwan. The most common mutations of beta-globin are point mutations, and six mutations account for over 90% of cases. Less than 5% of the cases with beta-globin gene deletion result in beta-thalassemia minor. The mutational type of the deletion is not clear in Taiwanese. We used polymerase chain reaction (PCR)-based methods to detect the breakpoint junctions of different deletional types of beta-thalassemia. In total, six cases of clinically suspected deletional type of beta-thalassemia were studied. The results showed that there were three types of deletions in these cases: two cases each for hereditary persistent fetal hemoglobinemia (HPFH) of the Southeast Asian (SEA) type, HPFH of the Yunnanese type, and gamma(G)+(gamma(A)deltabeta)(0)deletions, respectively. The clinical features of these deletional mutations are milder than the beta(o) types of the point mutation. The patients with compound heterozygous mutations of the point mutation and the deletional mutation are always transfusion independent.

Expression of DAZL protein in the human corpus luteum.

The DAZL gene and its homologues are required for the development of male and female germ cells in different species. However, their role in other aspects of human reproduction is not known. We have generated a polyclonal antibody to the DAZL protein and developed a sensitive standard curve quantitative-competitive-RT-PCR assay to characterize the expression of DAZL in the human corpus luteum (CL). DAZL transcripts are expressed in the CL, but the concentrations decreased with advancing luteal phase. In accordance with the mRNA data, DAZL protein was most abundant in the early phase CL. Immunohistochemical staining showed DAZL protein in the cytoplasm of granulosa-luteal cells. The distinct expression pattern of DAZL protein in the human CL may play an important role in the regulation of luteal function.

AZFa candidate gene deletions in Taiwanese patients with spermatogenic failure.

BACKGROUND AND PURPOSE: Deletions of the azoospermia factor subregion a (AZFa) genes in proximal Yq11 are not frequently reported. The majority of AZFa deletions are thought to be associated with more severe testicular phenotypes, such as Sertoli cell-only syndrome. There is a lack of data on AZFa gene deletions in East Asian populations. In this study, we investigated the deletion status of AZFa genes in Taiwanese men with spermatogenic failure. METHODS: One hundred and eighty-three consecutive men with severe oligozoospermia or non-obstructive azoospermia were enrolled in this study. Genomic DNA was extracted from peripheral blood samples and polymerase chain reaction (PCR) was performed using primers specific to four AZFa genes: AZFaT1, DFFRY, DBY, and UTY. Sequence-tagged site markers (sY740, sY630, sY86, sY85, sY87, sY709, and sY88) were used to define the position of deletions. One hundred and twenty fertile men with normal spermatogenesis were enrolled as controls. RESULTS: Of the 183 patients, two showed single AZFa gene deletions, resulting in an overall frequency of 1.1%. One of these two patients had DFFRY deletion and the other had DBY deletion; their testicular phenotypes were Sertoli cell-only syndrome and hypospermatogenesis, respectively. Neither patient had deletions extending from AZFa through AZFb or AZFc. CONCLUSION: Our results suggest that AZFa gene deletion is infrequent in Taiwanese patients with severe oligozoospermia or non-obstructive azoospermia.

Expression patterns and transcript concentrations of the autosomal DAZL gene in testes of azoospermic men.

The DAZ (Deleted in AZoospermia) gene cluster on the Y chromosome is a strong candidate for the azoospermia factor. The DAZ gene was derived from an autosomal homologue, DAZL (DAZ-Like). This study was designed to assess the functional role of DAZL in human spermatogenesis. The expression patterns and mRNA transcript levels of DAZL in the testes of 17 azoospermic men were therefore examined by immunohistochemical staining and quantitative competitive reverse transcription-polymerase chain reaction. DAZL protein was expressed in the cytoplasm of primary spermatocytes and weakly in spermatogonia. It was detected in the testicular tissues of all subjects with germ cells present. The copy number of the DAZL transcript in normal spermatogenesis (n = 4), hypospermatogenesis or maturation arrest (n = 6), and Sertoli cell-only syndrome (n = 7) ranged from 1.22 x 10(6) to 1.63 x 10(6) per ng of RNA, 1.19 x 10(5) to 2.82 x 10(5) per ng of RNA and 2.83 x 10(4) to 1.23 x 10(5) per ng of RNA respectively. DAZL transcripts were lower in men with spermatogenic failure, and a significant difference was found between the three groups (P < 0.0001). This study suggests that DAZL may play an important role in the human spermatogenic processes of both mitosis and meiosis.

Elastic properties of tendon measured by two different approaches.

Elastic properties of tendon were assessed by two different approaches. Six fresh bovine Achilles tendon specimens were used. The first approach directly measured Young's modulus along the transverse direction (E(perpendicular)) and the longitudinal direction (E(parallel)), using a cyclic compression-relaxation method. Young's moduli were derived based on the measured strain and stress values. The ratio of E(parallel): E(perpendicular) at smaller strains was around 4 and decreased to 0.6 approximately 1.1 at larger strains. The second approach assumed that tendons are transversely isotropic. Three observable second-order elastic stiffness constants (c(11), c(13) and c(33)) were obtained by sound speed measurements along various propagation directions. The measured elastic stiffness constants were also correlated with results from the first approach. It was shown that the transverse isotropy assumption was valid at small strains. However, a significant discrepancy existed between the two approaches. The discrepancy was primarily due to viscoelasticity associated with the first approach.

Magnetic-activated cell sorting (MACS) significantly decreases the hybridization efficiency of fluorescence in situ hybridization (FISH).

Fetal cells were enriched from maternal blood using density gradient centrifugation of Histopaque followed by magnetic-activated cell sorting (MACS) to select CD71-positive cells. For each specimen, cells partially purified by Histopaque were split into equal portions, and each portion was subjected to purification by MACS in parallel. Cells before and after MACS were subjected to dual-color fluorescence in situ hybridization (FISH) analysis with X- and Y-chromosome-specific probes. We found that the hybridization rates were decreased by approximately 10% after MACS based on duplicated analysis for each sample.

An effective strategy of using molecular testing to screen mentally retarded individuals for fragile X syndrome.

Fragile X syndrome (FXS) is the most common form of familial mental retardation (MR). It is caused by the expansion of the CGG repeat in the FMR1 gene on the X chromosome. To date, FXS is not treatable, but can be prevented by prenatal genetic examination. Identifying women who carry a full mutation or premutation FMR1 gene is thus very important, and can be done by tracing family members of FXS subjects. However, most of the FXS subjects in Taiwan as well as those in many other countries have not been identified. In this study the authors attempt to develop reliable and inexpensive tests suitable for a large-scale screen of subjects with MR for FXS. Together with their previous study, a total of 311 male and 160 female subjects with MR were screened with nonradioactive Southern blot assay using mixed deoxyribonucleic acid from three subjects of the same sex. From these subjects, nine male subjects and one female FXS subject were diagnosed. All male subjects were also screened with nonradioactive polymerase chain reaction (PCR). These nine male FXS subjects were also detected on the basis of PCR amplification failure. No false-negative results were discerned. The PCR procedure was simplified further by combining it with an analysis of a blood spot on filter paper, which is a much simpler and cheaper method for sample collection and DNA preparation. This method was then used to screen 104 boys with MR. Two of them were suspected, and later confirmed with Southern blot assay, as subjects with FXS. This study suggests that simple PCR combined with blood spot analysis could be a reliable, inexpensive test that is feasible for a large-scale screening of male subjects with MR for FXS. However, Southern blot assay with mixed deoxyribonucleic acid is appropriate for screening female subjects. Based on this strategy, most FXS subjects could be identified easily for further management.