The Role of Cilostazol, a Phosphodiesterase 3 Inhibitor, on Oocyte Maturation and Subsequent Pregnancy in Mice

It is important to identify effective contraceptive drugs that cause minimal disruption to physiological processes. Phosphodiesterase 3 (PDE3) inhibitors suppress meiosis in oocytes by decreasing the level of cAMP and blocking the extrusion of the first polar body. In this study, we tested the PDE3 inhibitor, cilostazol, as a potential contraceptive agent. The effects of cilostazol treatment in vitro and in vivo on the suppression of oocyte maturation in a mouse model were investigated. The results indicated that treatment with increasing concentrations of cilostazol led to a dose-dependent arrest in meiosis progression. The effective in vitro concentration was 1 µM and was 300 mg/kg in vivo. The effect of cilostazol was reversible. After removal of the drug, meiosis resumed and mouse oocytes matured in vitro, and showed normal chromosome alignment and spindle organization. After fertilization using an ICSI method, the oocytes showed normal morphology, fertilization rate, embryo cleavage, blastocyst formation, and number of viable pups when compared with controls. The offspring showed similar body weight and fertility. In vivo, the mice became infertile if the drug was injected sequentially, and became pregnant following discontinuation of cilostazol. More importantly, no side effects of cilostazol were observed in treated female mice as demonstrated by blood pressure and heart rate monitoring. It is concluded that cilostazol, a drug routinely used for intermittent claudication, can effectively inhibit oocyte maturation in vitro and in vivo, does not affect the developmental potential of oocytes following drug removal and has few side effects in female mice treated with this drug. These findings suggest that cilostazol may be a potential new contraceptive agent that may facilitate an efficacy and safety study of this drug

Antifungal susceptibility profiles and drug resistance mechanisms of clinical Candida duobushaemulonii isolates from China

Candida duobushaemulonii, type II Candida haemulonii complex, is closely related to Candida auris and capable of causing invasive and non-invasive infections in humans. Eleven strains of C. duobushaemulonii were collected from China Hospital Invasive Fungal Surveillance Net (CHIF-NET) and identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF), VITEK 2 Yeast Identification Card (YST), and internal transcribed spacer (ITS) sequencing. Whole genome sequencing of C. duobushaemulonii was done to determine their genotypes. Furthermore, C. duobushaemulonii strains were tested by Sensititre YeastOne™ and Clinical and Laboratory Institute (CLSI) broth microdilution panel for antifungal susceptibility. Three C. duobushaemulonii could not be identified by VITEK 2. All 11 isolates had high minimum inhibitory concentrations (MICs) to amphotericin B more than 2 μg/ml. One isolate showed a high MIC value of ≥64 μg/ml to 5-flucytosine. All isolates were wild type (WT) for triazoles and echinocandins. FUR1 variation may result in C. duobushaemulonii with high MIC to 5-flucytosine. Candida duobushaemulonii mainly infects patients with weakened immunity, and the amphotericin B resistance of these isolates might represent a challenge to clinical treatment

Type 2 Diabetes Variants Disrupt Function of SLC16A11 through Two Distinct Mechanisms

Type 2 diabetes (T2D) affects Latinos at twice the rate seen in populations of European descent. We recently identified a risk haplotype spanning SLC16A11 that explains ∼20% of the increased T2D prevalence in Mexico. Here, through genetic fine-mapping, we define a set of tightly linked variants likely to contain the causal allele(s). We show that variants on the T2D-associated haplotype have two distinct effects: (1) decreasing SLC16A11 expression in liver and (2) disrupting a key interaction with basigin, thereby reducing cell-surface localization. Both independent mechanisms reduce SLC16A11 function and suggest SLC16A11 is the causal gene at this locus. To gain insight into how SLC16A11 disruption impacts T2D risk, we demonstrate that SLC16A11 is a proton-coupled monocarboxylate transporter and that genetic perturbation of SLC16A11 induces changes in fatty acid and lipid metabolism that are associated with increased T2D risk. Our findings suggest that increasing SLC16A11 function could be therapeutically beneficial for T2D. Video Abstract [Figure presented] Keywords: type 2 diabetes (T2D); genetics; disease mechanism; SLC16A11; MCT11; solute carrier (SLC); monocarboxylates; fatty acid metabolism; lipid metabolism; precision medicin

The trans-ancestral genomic architecture of glycemic traits

Glycemic traits are used to diagnose and monitor type 2 diabetes and cardiometabolic health. To date, most genetic studies of glycemic traits have focused on individuals of European ancestry. Here we aggregated genome-wide association studies comprising up to 281,416 individuals without diabetes (30% non-European ancestry) for whom fasting glucose, 2-h glucose after an oral glucose challenge, glycated hemoglobin and fasting insulin data were available. Trans-ancestry and single-ancestry meta-analyses identified 242 loci (99 novel; P < 5 x 10(-8)), 80% of which had no significant evidence of between-ancestry heterogeneity. Analyses restricted to individuals of European ancestry with equivalent sample size would have led to 24 fewer new loci. Compared with single-ancestry analyses, equivalent-sized trans-ancestry fine-mapping reduced the number of estimated variants in 99% credible sets by a median of 37.5%. Genomic-feature, gene-expression and gene-set analyses revealed distinct biological signatures for each trait, highlighting different underlying biological pathways. Our results increase our understanding of diabetes pathophysiology by using trans-ancestry studies for improved power and resolution. A trans-ancestry meta-analysis of GWAS of glycemic traits in up to 281,416 individuals identifies 99 novel loci, of which one quarter was found due to the multi-ancestry approach, which also improves fine-mapping of credible variant sets.Peer reviewe

Genetic Drivers of Heterogeneity in Type 2 Diabetes Pathophysiology

Type 2 diabetes (T2D) is a heterogeneous disease that develops through diverse pathophysiological processes1,2 and molecular mechanisms that are often specific to cell type3,4. Here, to characterize the genetic contribution to these processes across ancestry groups, we aggregate genome-wide association study data from 2,535,601 individuals (39.7% not of European ancestry), including 428,452 cases of T2D. We identify 1,289 independent association signals at genome-wide significance (P \u3c 5 × 10-8) that map to 611 loci, of which 145 loci are, to our knowledge, previously unreported. We define eight non-overlapping clusters of T2D signals that are characterized by distinct profiles of cardiometabolic trait associations. These clusters are differentially enriched for cell-type-specific regions of open chromatin, including pancreatic islets, adipocytes, endothelial cells and enteroendocrine cells. We build cluster-specific partitioned polygenic scores5 in a further 279,552 individuals of diverse ancestry, including 30,288 cases of T2D, and test their association with T2D-related vascular outcomes. Cluster-specific partitioned polygenic scores are associated with coronary artery disease, peripheral artery disease and end-stage diabetic nephropathy across ancestry groups, highlighting the importance of obesity-related processes in the development of vascular outcomes. Our findings show the value of integrating multi-ancestry genome-wide association study data with single-cell epigenomics to disentangle the aetiological heterogeneity that drives the development and progression of T2D. This might offer a route to optimize global access to genetically informed diabetes care

Genetic drivers of heterogeneity in type 2 diabetes pathophysiology

Type 2 diabetes (T2D) is a heterogeneous disease that develops through diverse pathophysiological processes1,2 and molecular mechanisms that are often specific to cell type3,4. Here, to characterize the genetic contribution to these processes across ancestry groups, we aggregate genome-wide association study data from 2,535,601 individuals (39.7% not of European ancestry), including 428,452 cases of T2D. We identify 1,289 independent association signals at genome-wide significance (P &lt; 5 × 10-8) that map to 611 loci, of which 145 loci are, to our knowledge, previously unreported. We define eight non-overlapping clusters of T2D signals that are characterized by distinct profiles of cardiometabolic trait associations. These clusters are differentially enriched for cell-type-specific regions of open chromatin, including pancreatic islets, adipocytes, endothelial cells and enteroendocrine cells. We build cluster-specific partitioned polygenic scores5 in a further 279,552 individuals of diverse ancestry, including 30,288 cases of T2D, and test their association with T2D-related vascular outcomes. Cluster-specific partitioned polygenic scores are associated with coronary artery disease, peripheral artery disease and end-stage diabetic nephropathy across ancestry groups, highlighting the importance of obesity-related processes in the development of vascular outcomes. Our findings show the value of integrating multi-ancestry genome-wide association study data with single-cell epigenomics to disentangle the aetiological heterogeneity that drives the development and progression of T2D. This might offer a route to optimize global access to genetically informed diabetes care.</p

Riociguat treatment in patients with chronic thromboembolic pulmonary hypertension: Final safety data from the EXPERT registry

Objective: The soluble guanylate cyclase stimulator riociguat is approved for the treatment of adult patients with pulmonary arterial hypertension (PAH) and inoperable or persistent/recurrent chronic thromboembolic pulmonary hypertension (CTEPH) following Phase

SV40 T/t-common polypeptide inhibits angiogenesis and enhances the cytotoxic activity of chemotherapeutic agents in human HER2-overexpressing tumors

HER2 致癌基因在許多人類癌細胞 (特別是卵巢癌和乳癌) 中有大量表現的情形。研究發現HER2會促進癌細胞生長、存活、血管新生 (angiogenesis)、癌細胞轉移 (metastasis)、抑制癌細胞對化療藥物的感受性 (chemosensitivity) 和降低生命存活率，若在HER2大量表現的癌細胞中抑制HER2的表現則會降低癌細胞生長、存活、血管新生、增加癌細胞對化療藥物感受性和增加生命存活率。我們先前證明了猴病毒四十型（SV40）的T/t-common polypeptide在HER2大量表現的癌細胞會透過抑制HER2的表現進而抑制癌細胞形成腫瘤的能力，此外也可使HER2大量表現的人類卵巢癌和乳癌細胞進行細胞凋亡(apoptosis)。本論文第一部分探討T/t-common在具有HER2大量表現的人類卵巢癌細胞SK-OV-3中是否會抑制血管新生。利用具有T/t-common 表現的SK-OV-3 細胞培養液(conditioned medium) 發現會抑制內皮細胞的移動 (migration)、血管生成 (tube formation) 和動物體內血管生成 (microvessel formation)，證明了T/t-common 可抑制SK-OV-3誘發血管新生的能力。若將HER2基因重新送入可表現T/t-common的SK-OV-3細胞 (T/t-common stable clones)中並使HER2再大量表現；收集細胞培養液進行實驗，發現HER2會降低T/t-common抑制內皮細胞移動和血管生成的能力，因此T/t-common抑制血管新生能力是透過抑制了HER2的表現。T/t-common在SK-OV-3中會抑制一些血管新生因子 (proangiogenic factors) 例如（VEGF-A、IL-8、bFGF、uPA、MMP-2）的表現和促進一些抑制血管新生因子 (antiangiogenic factors) 例如（TSP-1、TIMP-1）的表現。我們也進一步証明T/t-common在SK-OV-3中是透過抑制 ERK1/2活性進而抑制轉錄因子HIF-1α 和血管新生因子VEGF-A的表現，此外T/t-common可促進p38活性進而增加抗血管新生因子TSP-1的表現，因此T/t-common可透過HER2-ERK1/2-HIF-1α-VEGF-A和HER2-p38-TSP-1訊息傳遞路徑來抑制HER2誘發血管新生能力。將HER2大量表現的卵巢癌細胞SK-OV-3種植在免疫缺陷老鼠NOD/SCID並以T/t-common治療，發現T/t-common在NOD/SCID老鼠可抑制HER2大量表現腫瘤的血管新生和腫瘤生長。因此利用T/t-common做基因治療可能是一種治療HER2大量表現癌症的新方法。 本論文第二部份則探討T/t-common在具抗藥性的HER2大量表現的癌細胞中，是否能增強癌細胞對化療藥物cisplatin (CDDP)及doxorubicin (DXR)的感受性，並進一步探討其增強化療藥物療效的機制。結果顯示T/t-common可特異性地加強CDDP或DXR 對HER2大量表現癌細胞的毒殺能力，並經由增加Caspase 3的活性誘發癌細胞進行細胞凋亡而增加化療藥物療效。當HER2大量表現的癌細胞同時以T/t-common 和CDDP或DXR作用下，發現T/t-common可抑制ERK1/2的活性和促進JNK的活性，並藉此抑制Bcl-2和Bcl-XL的表現，因此T/t-common可能是經由調控上述細胞凋亡因子進而增強CDDP或DXR誘發HER2大量表現癌細胞細胞凋亡。最後將HER2大量表現的卵巢癌細胞SK-OV-3.ip1植入免疫缺陷老鼠NOD/SCID，並以T/t-common和化療藥物DXR一同治療，發現腫瘤體積比任一單獨試劑治療還小。因為T/t-common可增強CDDP或DXR毒殺HER2大量表現癌細胞的能力，所以T/t-common與CDDP或DXR的合同療法也許可克服HER2大量表現癌細胞對化療藥物的抗藥性，因此是一種具有潛力的治療HER2大量表現癌細胞的新方法。Overexpression of HER2 has been frequently detected in many types of human cancer, most notably breast and ovarian cancers. HER2 overexpression is associated with increased angiogenesis, increased metastasis, increased chemoresistance and reduced survival. Inhibition of HER2 in HER2-overexpressing cancers can lead to reduced angiogenesis, reduced chemoresistance and improved survival. Previously, we reported that Simian Virus 40 T/t-common polypeptide can inhibit HER2, suppress the tumorigenic potential of HER2-overexpressing cancer cells and specifically induce apoptosis in HER2-overexpressing human cancer cell lines. In the part I of this thesis, we further explored the effect of T/t-common on tumor angiogenesis. We found that T/t-common could inhibit the ability of HER2-overexpressing cancer cells, but not low HER2-expressing cancer cells, to induce the migration and tube formation of endothelial cells. Reexpression of HER2 could block T/t-common’s activity to inhibit the ability of HER2-overexpressing cancer cells to induce the migration and tube formation of endothelial cells. We further tested whether T/t-common could affect HER2-overexpressing cancer cells to express and secrete angiogenic and anti-angiogenic factors. We found that T/t-common could inhibit the expression of proangiogenic factor VEGF-A, IL-8, bFGF, uPA and MMP-2, and induce the expression of anti-angiogenic factor TSP-1and TIMP-1 in HER2-overexpressing human SK-OV-3 ovarian cancer cells. T/t-common was found to be able to repress the expression of HIF-1α, a transcription factor required for VEGF-A expression, through inhibiting HER2-mediated ERK1/2 activity. Moreover, T/t-common was found to be able to activate p38 pathway, leading to up-regulation of TSP-1. Through regulating HER2-ERK1/2-HIF-1α-VEGF-A and HER2-p38-TSP-1 signaling pathways in HER2-overexpressing cancer cells, T/t-common could inhibit these cancer cells to recruit endothelial cells, leading to inhibition of angiogenesis in HER2-overexpressing tumors. Two experiments described below demonstrated that T/t-common indeed can inhibit tumor angiogenesis. First, in Matrigel plug assays, we demonstrated that conditioned medium from T/t-common-transduced SK-OV-3 cancer cells had lower ability to induce blood vessel formation in Matrigel plugs than that from control SK-OV-3 cells. Second, infection of adenovirus carrying T/t-common gene could lead to inhibition of the growth and microvessel formation of SK-OV-3 tumors in NOD/SCID mice model. Taken together, the above suggest that T/t-common had the potential to be developed as a new antiangiogenic agent specific for treating HER2-overexpressing ovarian cancers. In the part II of this thesis, we investigated whether T/t-common could enhance the sensitivity of HER2-overexpressing human cancer cells to chemotherapeutic agents such as cisplatin (CDDP) and doxorubicin (DXR). We found that T/t-common could specifically enhance the sensitivity of HER2-overexpressing human cancer cells, but not that of HER2 low-expressing human cancer cells, to CDDP and DXR. T/t-common could specifically enhance CDDP- or DXR-induced apoptosis by activating caspase-3 activity in HER2-overexpressing cancer cells. T/t-common expression led to down-regulation of Bcl-2 and Bcl-XL, inhibition of ERK activity, and activation of JNK activity in CDDP- or DXR-treated HER2-overexpressing BT-474 cancer cells. This modulation of apoptosis regulatory molecules by T/t-common may contribute to its ability to enhance CDDP- or DXR-induced apoptosis in HER2-overexpressing cancer cells. Finally, we showed that T/t-common could enhance the antitumor activity of DXR on HER2-overexpressing SK-OV-3.ip1 tumor in NOD/SCID mice. Together, these data demonstrated that T/t-common could sensitize HER2-overexpressing cancer cells to chemotherapeutic agents CDDP or DXR, and suggest that combination therapy using T/t-common gene and chemotherapeutic agents (such as CDDP or DXR) may overcome tumor chemoresistance and thus provide a new approach for treatment of HER2-overexpressing cancers

Exploring Role Behavior in Restaurant by Grey Model and Grey Structural Model

Based on GM (0,N) grey model and grey structure model in grey system theory, this study takes Chinese restaurant in tourist hotel as a case to analyze service role behavior. A total of 241 questionnaires were collected to calculate index weighted coefficients, and then 12 experts carried out an investigation to construct clusters. There were 12 dimensions of professional competencies, and a total of 50 indicator factors were analyzed for role behavior in a restaurant. According to the results, there are three role behaviors for service staff in Chinese restaurants: supportive, interactive, and integrative role behaviors. In theory, this reinterprets the meaning of catering service competencies and defines the role types of catering service staff. In practical applications, restaurant managers could apply this result to help service staff to understand their current role, in order to reduce their role pressure and to increase their job satisfaction and performance

Development of a Performance-Based Measure of Executive Functions in Patients with Schizophrenia.

A performance-based measure for assessing executive functions (EF) is useful to understand patients' real life performance of EF. This study aimed to develop a performance-based measure of executive functions (PEF) based on the Lezak model and to examine psychometric properties (i.e., unidimensionality and reliability) of the PEF using Rasch analysis in patients with schizophrenia. We developed the PEF in three phases: (1) designing the preliminary version of PEF; (2) consultation with experts, cognitive interviews with patients, and pilot tests on patients to revise the preliminary PEF; (3) establishment of the final version of the PEF and examination of unidimensionality and Rasch reliability. Two hundred patients were assessed using the revised PEF. After deleting items which did not satisfy the Rasch model's expectations, the final version of the PEF contained 1 practice item and 13 test items for assessing the four domains of EF (i.e., volition, planning, purposive action, and effective performance). For unidimensional and multidimensional Rasch analyses, the 4 domains showed good reliability (i.e., 0.77-0.85 and 0.87-0.90, respectively). Our results showed that the PEF had satisfactory unidimensionality and Rasch reliability. Therefore, clinicians and researchers could use the PEF to assess the four domains of EF in patients with schizophrenia