Investigation of Hepatoprotective Activity of Induced Pluripotent Stem Cells in the Mouse Model of Liver Injury

To date liver transplantation is the only effective treatment for end-stage liver diseases. Considering the potential of pluripotency and differentiation into tridermal lineages, induced pluripotent stem cells (iPSCs) may serve as an alternative of cell-based therapy. Herein, we investigated the effect of iPSC transplantation on thioacetamide- (TAA-) induced acute/fulminant hepatic failure (AHF) in mice. Firstly, we demonstrated that iPSCs had the capacity to differentiate into hepatocyte-like cells (iPSC-Heps) that expressed various hepatic markers, including albumin, α-fetoprotein, and hepatocyte nuclear factor-3β, and exhibited biological functions. Intravenous transplantation of iPSCs effectively reduced the hepatic necrotic area, improved liver functions and motor activity, and rescued TAA-treated mice from lethal AHF. 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate cell labeling revealed that iPSCs potentially mobilized to the damaged liver area. Taken together, iPSCs can effectively rescue experimental AHF and represent a potentially favorable cell source of cell-based therapy

Genome-Wide Gene Expression Analysis Implicates the Immune Response and Lymphangiogenesis in the Pathogenesis of Fetal Chylothorax

Fetal chylothorax (FC) is a rare condition characterized by lymphocyte-rich pleural effusion. Although its pathogenesis remains elusive, it may involve inflammation, since there are increased concentrations of proinflammatory mediators in pleural fluids. Only a few hereditary lymphedema-associated gene loci, e.g. VEGFR3, ITGA9 and PTPN11, were detected in human fetuses with this condition; these cases had a poorer prognosis, due to defective lymphangiogenesis. In the present study, genome-wide gene expression analysis was conducted, comparing pleural and ascitic fluids in three hydropic fetuses, one with and two without the ITGA9 mutation. One fetus (the index case), from a dizygotic pregnancy (the cotwin was unaffected), received antenatal OK-432 pleurodesis and survived beyond the neonatal stage, despite having the ITGA9 mutation. Genes and pathways involved in the immune response were universally up-regulated in fetal pleural fluids compared to those in ascitic fluids. Furthermore, genes involved in the lymphangiogenesis pathway were down-regulated in fetal pleural fluids (compared to ascitic fluid), but following OK-432 pleurodesis, they were up-regulated. Expression of ITGA9 was concordant with overall trends of lymphangiogenesis. In conclusion, we inferred that both the immune response and lymphangiogenesis were implicated in the pathogenesis of fetal chylothorax. Furthermore, genome-wide gene expression microarray analysis may facilitate personalized medicine by selecting the most appropriate treatment, according to the specific circumstances of the patient, for this rare, but heterogeneous disease

Lapatinib Induces Autophagy, Apoptosis and Megakaryocytic Differentiation in Chronic Myelogenous Leukemia K562 Cells

Lapatinib is an oral, small-molecule, dual tyrosine kinase inhibitor of epidermal growth factor receptors (EGFR, or ErbB/Her) in solid tumors. Little is known about the effect of lapatinib on leukemia. Using human chronic myelogenous leukemia (CML) K562 cells as an experimental model, we found that lapatinib simultaneously induced morphological changes resembling apoptosis, autophagy, and megakaryocytic differentiation. Lapatinib-induced apoptosis was accompanied by a decrease in mitochondrial transmembrane potential and was attenuated by the pancaspase inhibitor z-VAD-fmk, indicating a mitochondria-mediated and caspase-dependent pathway. Lapatinib-induced autophagic cell death was verified by LC3-II conversion, and upregulation of Beclin-1. Further, autophagy inhibitor 3-methyladenine as well as autophagy-related proteins Beclin-1 (ATG6), ATG7, and ATG5 shRNA knockdown rescued the cells from lapatinib-induced growth inhibition. A moderate number of lapatinib-treated K562 cells exhibited features of megakaryocytic differentiation. In summary, lapatinib inhibited viability and induced multiple cellular events including apoptosis, autophagic cell death, and megakaryocytic differentiation in human CML K562 cells. This distinct activity of lapatinib against CML cells suggests potential for lapatinib as a therapeutic agent for treatment of CML. Further validation of lapatinib activity in vivo is warranted

Genome-wide association study of lung adenocarcinoma in East Asia and comparison with a European population.

Lung adenocarcinoma is the most common type of lung cancer. Known risk variants explain only a small fraction of lung adenocarcinoma heritability. Here, we conducted a two-stage genome-wide association study of lung adenocarcinoma of East Asian ancestry (21,658 cases and 150,676 controls; 54.5% never-smokers) and identified 12 novel susceptibility variants, bringing the total number to 28 at 25 independent loci. Transcriptome-wide association analyses together with colocalization studies using a Taiwanese lung expression quantitative trait loci dataset (n = 115) identified novel candidate genes, including FADS1 at 11q12 and ELF5 at 11p13. In a multi-ancestry meta-analysis of East Asian and European studies, four loci were identified at 2p11, 4q32, 16q23, and 18q12. At the same time, most of our findings in East Asian populations showed no evidence of association in European populations. In our studies drawn from East Asian populations, a polygenic risk score based on the 25 loci had a stronger association in never-smokers vs. individuals with a history of smoking (Pinteraction = 0.0058). These findings provide new insights into the etiology of lung adenocarcinoma in individuals from East Asian populations, which could be important in developing translational applications

Genome-wide association study of lung adenocarcinoma in East Asia and comparison with a European population

Lung adenocarcinoma is the most common type of lung cancer. Known risk variants explain only a small fraction of lung adenocarcinoma heritability. Here, we conducted a two-stage genome-wide association study of lung adenocarcinoma of East Asian ancestry (21,658 cases and 150,676 controls; 54.5% never-smokers) and identified 12 novel susceptibility variants, bringing the total number to 28 at 25 independent loci. Transcriptome-wide association analyses together with colocalization studies using a Taiwanese lung expression quantitative trait loci dataset (n = 115) identified novel candidate genes, including FADS1 at 11q12 and ELF5 at 11p13. In a multi-ancestry meta-analysis of East Asian and European studies, four loci were identified at 2p11, 4q32, 16q23, and 18q12. At the same time, most of our findings in East Asian populations showed no evidence of association in European populations. In our studies drawn from East Asian populations, a polygenic risk score based on the 25 loci had a stronger association in never-smokers vs. individuals with a history of smoking (P interaction  = 0.0058). These findings provide new insights into the etiology of lung adenocarcinoma in individuals from East Asian populations, which could be important in developing translational applications

Exploring the Development of Professional Competence in Nurse Practitioners in Taiwan

Background and purpose: Since the introduction of a certification examination in 2006, 6,414 nurse practitioners have been certified. Few studies of the professional competence of these nurse practitioners have been conducted, and none have been conducted in the past 8 years. The aim of this study was evaluate the development of professional competence of nurse practitioners in Taiwan. Methods: In-depth interviews of 11 nurse practitioners were conducted. Data were collected by purposive sampling. Theoretical sampling and grounded theory techniques were used in this qualitative analysis. Results: The development of professional competence in nurse practitioners included three stages, self-development motivation, professional development working as a nurse practitioner, and experiencing a sense of achievement as a nurse practitioner. The core category was identified as being a mature, competent nurse practitioner. Discussion: During their clinical practice, nurse practitioners should be informed well in advance of inservice training. Development of learning guidelines is recommended to overcome a lack of continuing education in practical nursing areas. Most nurse practitioners stated that they did not receive adequate pre-employment training and that they learned to adapt to practice on their own. Future efforts should include implementation of online courses conducted by experienced nurse practitioners discussing the essential knowledge and skills that individuals should have before beginning work a nurse practitioner or following their transition to practice. The goal would be to help in the development of professional competence

Tantao Shouci Nao Zhongfeng Bingren Chuqi zhi Tiaoshi Guocheng

背景　腦中風是全世界主要死因之一，在台灣位居十大死因之第三位，也是導致成人失能的主因，對初次中風病人的衝擊很大，其影響包含生理、心理及社會層面，目前國內對於探討首次中風病人於初期的調適過程較闕如。目的　本研究欲探索首次腦中風病人初期之調適過程。方法　採質性研究，以立意取樣，隨研究分析過程中，隨後進行理論性抽樣，共訪談十二位初次腦中風病人，並以紮根理論為研究方法及分析的方式，使用譯碼程序進行資料分析及統整。結果　本研究發現，首次中風病人初期的調適過程包含兩個範疇、六個次範疇及一個核心範疇，其分別為「中風的衝擊」：㈠失去身體自主權；㈡喪失自我價值；㈢難以承受，產生社交隔離。以及「中風後的因應」：㈠積極復健，重獲自主權；㈡家人支持，努力不成為負擔；㈢由苦難中昇華，轉趨正向概念。核心範疇為「面對中風，懷抱希望，人生就此改變」。結論／實務應用　研究結果呈現腦中風病人在初期調適過程的原貌，藉此提供臨床護理人員照護此類病人之參考，建議臨床上除了持續關注腦中風病人的初期調適外，更應針對病人在中風後因應過程中給予所需的幫助，並視其需求給予衛教及心理支持

Pigment Epithelium Derived Factor Peptide Protects Murine Hepatocytes from Carbon Tetrachloride-Induced Injury

<div><p>Fibrogenesis is induced by repeated injury to the liver and reactive regeneration and leads eventually to liver cirrhosis. Pigment epithelium derived factor (PEDF) has been shown to prevent liver fibrosis induced by carbon tetrachloride (CCl₄). A 44 amino acid domain of PEDF (44-mer) was found to have a protective effect against various insults to several cell types. In this study, we investigated the capability of synthetic 44-mer to protect against liver injury in mice and in primary cultured hepatocytes. Acute liver injury, induced by CCl₄, was evident from histological changes, such as cell necrosis, inflammation and apoptosis, and a concomitant reduction of glutathione (GSH) and GSH redox enzyme activities in the liver. Intraperitoneal injection of the 44-mer into CCl₄-treated mice abolished the induction of AST and ALT and markedly reduced histological signs of liver injury. The 44-mer treatment can reduce hepatic oxidative stress as evident from lower levels of lipid hydroperoxide, and higher levels of GSH. CCl₄ caused a reduction of Bcl-xL, PEDF and PPARγ, which was markedly restored by the 44-mer treatment. Consequently, the 44-mer suppressed liver fibrosis induced by repeated CCl₄ injury. Furthermore, our observations in primary culture of rat hepatocytes showed that PEDF and the 44-mer protected primary rat hepatocytes against apoptosis induced by serum deprivation and TGF-β1. PEDF/44-mer induced cell protective STAT3 phosphorylation. Pharmacological STAT3 inhibition prevented the antiapoptotic action of PEDF/44-mer. Among several PEDF receptor candidates that may be responsible for hepatocyte protection, we demonstrated that PNPLA2 was essential for PEDF/44-mer-mediated STAT3 phosphorylation and antiapoptotic activity by using siRNA to selectively knockdown PNPLA2. In conclusion, the PEDF 44-mer protects hepatocytes from single and repeated CCl₄ injury. This protective effect may stem from strengthening the counter oxidative stress capacity and induction of hepatoprotective factors.</p></div

Effect of STAT3 inhibitor and PEDF receptor siRNA on hepatocyte apoptosis induced by serum deprivation.

<p>(A) Inhibitor of STAT3 prevents the induction of PEDF by the 44-mer. Hepatocytes were pretreated with STAT3 inhibitor or ERK inhibitor (PD98059) or PPARγ antagonist (GW9662) for 2 h and then treated with the 44-mer for 24 h. To examine the role of PEDF receptor on antiapoptotic effect of the 44-mer, hepatocytes were transfected with the indicated siRNA. Two days later, the hepatocytes were starved of serum and exposed to the 44-mer for further 24 h. Subsequently, apoptosis was determined by TUNEL assay. Graphs represent means ± SE (n = 4). *<i>P</i><0.001 versus 44-mer-treated cell. **<i>P</i> < 0.05 relative to the cells pretreated with control siRNA/44-mer. (B) The 44-mer induces STAT3 phosphorylation. Hepatocytes were treated with the 44-mer for the indicated time periods and phosphorylated STAT3 were detected by western blot analysis. Representative immunoblots and densitometric analysis are shown as the mean ± SE (n = 3). *<i>P</i> < 0.001 versus untreated cells (time 0). (C-E) PNPLA2 siRNA reduces the PEDF/44-mer-induced STAT3 phosphorylation. Hepatocytes were transfected with control siRNA (lanes 2 and 3) or PEDF receptor specific siRNA (PNPLA2, LRP6 or LR; lanes 4 and 5) as indicated. Mock indicates cells treated with transfection reagent. PEDF receptor levels were normalized to the β-actin. Numbers below the blots refer to a densitometric measure expressed as a fold of mock control. To examine p-STAT3 levels, hepatocytes were treated with PEDF or 44-mer for 10 min and then lysed. Total and phosphorylated STAT3 protein levels were examined by western blotting. p-STAT3 was normalized to the STAT3. Graphs represent mean ± SE of 3 different experiments. *<i>P</i> < 0.05 versus Cont siRNA/PEDF-treated cells. **<i>P</i> < 0.05 versus Cont siRNA/44-mer-treated cells.</p