BACKGROUND:We previously demonstrated that maternal exposure to di-n-butyl phthalate (DBP) induces dysplasia of the kidney in newborn male offspring and renal fibrosis in adults. But the underlying mechanisms remain elusive. Fgf10/Fgfr2 and androgen receptor (AR) are known to be important for renal development. We therefore investigated whether these genes are involved in DBP-induced renal fibrosis. MATERIALS AND METHODS:Using Sprague-Dawley rats and rat renal proximal tubular cells (NRK52E), we determined the potential involvement of Fgf10, Fgfr2 and AR in DBP-induced renal fibrosis. RESULTS:We found that maternal exposure to DBP induces renal fibrosis in adult male offspring. A lower serum testosterone concentration and reduced expression of Fgf10, Fgfr2 and AR were detected in these animals. These was a trend toward lower expression of Fgf10, Fgfr2 and AR in NRK52E cells subjected to DBP exposure. Furthermore, higher expression levels of TGF-β and α-SMA were observed in abnormal renal tissue and DBP-treated NRK52E cells. CONCLUSION:Our findings suggest the potential involvement of Fgf10/Fgfr2 and AR in renal fibrosis of adult male rat offspring induced by prenatal exposure to DBP. The anti-androgenic effects of DBP might play an important role in this pathological process.

To investigate the effect of heat shock pretreatment on apoptosis and mitochondrial metallothionein (MT) expression in rat cardiomyocytes. In vitro cultured H9C2 cells were randomly divided into three groups: control, hydrogen peroxide (H2O2) injury, and H2O2 injury after heat shock pretreatment (n = 6 per group). Cardiomyocyte apoptosis and caspase-3 activity were assayed after treatment. Mitochondrial cytochrome (cyt) c and MT expression was assayed by Western blotting. Compared with the control group, the H2O2 injury group had a growing number of apoptotic cardiomyocytes (P < 0.01) and significantly elevated caspase-3 activity (P < 0.01) with markedly increased mitochondrial cyt c and MT expression (P < 0.01). After heat shock pretreatment, the numbers of apoptotic and necrotic cardiomyocytes (P < 0.01) and the caspase-3 activity significantly declined (P < 0.01), while mitochondrial cyt c and MT expression continued to increase (P < 0.01) compared with the H2O2 injury group. Heat shock pretreatment inhibits cardiomyocyte apoptosis, which may have a protective effect on cardiomyocytes by increasing the expression of myocardial protective MT and reducing the release of mitochondrial cyt c.

Objective To observe the changes of lymphocyte subsets of peripheral blood in 2 common oral mucosa diseases,recurrent aphthous ulcer (RAU ) and oral lichen planus (OLP ),and to investigate their immunological pathogenesis.Methods By flow cytometry,the lymphocyte subsets (total T cells,CD4 + T cells,CD8 +T cells, CD4 +/CD8,NK cells and B cells)in the peripheral blood of patients with RAU (105 cases),OLP (70 cases)and healthy controls (65 cases).The difference was analyzed comparatively between the groups.Results Compared with healthy controls,total T cells and CD8 +T cells in RAU group increased (P<0.05,P<0.01),but CD4 +T cells, CD4 +/CD8 +and B cells decreased in RAU group (P<0.01,P<0.001,P<0.05).In addition,CD8 +T cells in OLP group were lower than those in control group (P<0.05).The total T cells and CD8 +T cells in RAU group were higher than those in OLP group (P<0.01,P<0.001),while CD4 +T cells,CD4 +/CD8 +and NK cells in RAU group were significantly lower (P<0.01,P<0.001,P<0.05).Conclusions Owing to the imbalanced lymphocyte subset percentages,the cellular immune function of patients with RAU or OLP is often found to be abnormal,but their immune states are not completely same.So,the application of selective immunologic intervention may be the promising therapeutic measure to those oral mucosa diseases.

Chronic heart failure (CHF) is the final stage of various heart diseases, and is increasingly recognized as a major health problem in the elderly. Previous studies demonstrated that B‑type‑natriuretic peptide (BNP) is an established biomarker of CHF. Furthermore, BNP also regulates cell proliferation, differentiation and apoptosis. Recent evidence has revealed that BNP affects myocardial cell apoptosis during myocardial ischemia‑reperfusion injury. Long non‑coding RNAs (lncRNAs) are emerging as novel molecular compounds involved in gene regulation, and have important roles in numerous human diseases. However, the mechanism underlying the BNP and lncRNA-induced regulation of myocardial cell apoptosis remains to be elucidated. The present study reported that lncRNA LSINCT5, upregulated by BNP, is able to regulate myocardial cell apoptosis via the activation of the caspase‑1/interleukin (IL)‑1β signaling pathway. BNP-induced apoptosis of HCM cells was observed using flow cytometry, and involved caspase‑1. In addition, expression profiling using a human lncRNA polymerase chain reaction array revealed that LSINCT5 was highly expressed in BNP-treated myocardial cells, as compared with untreated cells. The role of lncRNA LSINCT5 in HCM cell apoptosis was also investigated. The results of the present study indicated that LSINCT5 silencing by small interfering RNA inhibits caspase‑1/IL‑1β signaling, and suppresses apoptosis in BNP-treated HCM cells. Therefore, high expression levels of BNP promote the apoptosis of myocardial cells through the lncRNA LSINCT5 mediator, which activates the caspase‑1/IL‑1β signaling pathway. These findings uncovered a novel pathogenic mechanism, and provided a potential therapeutic target for CHF.

目的:研究腹型肥胖对急性冠脉综合征(ACS)患者冠状动脉病变严重程度的影响.方法:选择2012年4月至2013年4月在我院接受治疗的ACS患者120例,根据腰围身高比(RWH)将患者分为无腹型肥胖者60例(对照组,RWH＜0.5)及腹型肥胖者60例(观察组,RWH＜0.5).测量所有患者的基本参数,计算RWH,利用冠脉造影判断冠脉病变的支数和程度,根据Gensini评分法对冠脉造影结果进行评价,分析冠脉病变范围、Gensini积分和RWH的相关性.结果:观察组的收缩压(SBP)、空腹血糖(FPG)、总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL-C)及Gensini积分水平均显著高于对照组,差异均有统计学意义(均P＜0.05).观察组的高密度脂蛋白(HDL-C)水平显著低于对照组,差异有统计学意义(P＜0.05).双支病变的RWH及Gensini积分水平显著高于单支病变,三支病变的RWH及Gensini积分水平显著高于单支病变及双支病变,差异均有统计学意义(均P＜0.05).根据Spearman法分析相关性可知,冠脉病变范围、Gensini积分和RWH均呈正相关(r=0.635,0.739;P=0.000,0.000).结论:ACS患者RWH水平增高与冠脉病变的严重程度关系密切,有效控制腹型肥胖对于降低心血管类疾病发病率以及降低冠脉病变的程度具有重要意义.

Uveal melanoma is the most serious primary intraocular malignancy in adults. Traditional approaches to treating uveal melanoma, including surgery, radiotherapy and chemotherapy, are insufficient. The oncolytic adenovirus H101 specifically replicates in and kills p53-mutant tumour cells while sparing normal cells and has been approved for use by the China Food and Drug Administration. To investigate the effect of treating uveal melanoma with H101, we infected specific uveal melanoma cell lines in vitro and observed significant cell growth inhibition, apoptosis and cell cycle arrest; the treatment was innocuous in the ARPE-19 normal cell line. In vivo, an intratumoural injection of H101 inhibited tumour growth and prolonged animal survival in the SP6.5 tumour xenograft mouse model. These results indicated that viral therapy using the oncolytic adenovirus H101 is feasible and promising for the treatment of uveal melanoma.

Uveal melanoma (UM) is the most common primary intraocular malignancy and the leading potentially fatal primary intraocular disease in adults. Melanoma antigen recognized by T-cells (MART-1) has been studied extensively as a clinically important diagnostic marker for melanoma, however, its biological function remains unclear. In the present study, the UM cell line SP6.5, which showed a high level of MART-1 expression, was subjected to small interfering RNA-mediated silencing of MART-1. Silencing of MART-1 expression increased the migration ability of SP6.5 cells and down-regulated the expression of the metastasis suppressor NM23. Our results suggest that MART-1 is a candidate target for the development of therapeutic strategies for UM and in particular for the suppression of metastasis associated with this malignancy.

Purpose.: Westudied the mechanism by which complete posterior vitreous detachment by enzyme-induced vitreolysis alleviates the progression of diabetic retinopathy. Methods.: We enrolled 50 diabetic rats and 20 normal control rats in this study. The right eyes of these diabetic rats were treated with a hyaluronidase (5 U) plus plasmin (0.25 U) by intravitreous injection (ET group), while the left eyes of diabetic rats (DR group) and eyes of the normal rats (NC group) were injected with balanced saline. Eight months after intravitreous injection, the oxygen concentration in the vitreous was measured, and the expression levels of hypoxia inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), pigment epithelium-derived factor (PEDF), and basic fibroblast growth factor (bFGF) in retinas were determined by real-time PCR and Western blot. Clinical observation, visual electrophysiology tests, and scanning electron microscopy (SEM) also were performed on the rats. Results.: SEM results showed that the surface of retinas in the ET group had little vitreous cortex and the inner limiting membrane could be observed. The oxygen concentration of the midvitreous was higher in the ET group than other groups. A significantly high expression of HIF-1α, VEGF, and bFGF was detected in the retinas of the DR group compared to the ET and NC groups. In visual electrophysiology tests, amplitude decline and peak time latency were found in diabetic rats, and changes in the DR group were more obvious than in the ET group. Conclusions.: Enzyme-induced vitreolysis can increase oxygen concentration in the vitreous, with reduced expression of HIF-1α, VEGF, and bFGF, and increased expression of PEDF in the retinas, thus alleviating the progression of diabetic retinopathy.

Maintaining the functional integrity of mitochondria is crucial for cell function, signal transduction and overall cell activities. Mitochondrial dysfunctions may alter energy metabolism and in many cases are associated with neurological diseases. Recent studies have reported that mutations in dehydrogenase E1 and transketolase domain‐containing 1 ( DHTKD1 ), a mitochondrial protein encoding gene, could cause neurological abnormalities. However, the function of DHTKD1 in mitochondria remains unknown. Here, we report a strong correlation of DHTKD1 expression level with ATP production, revealing the fact that DHTKD1 plays a critical role in energy production in mitochondria. Moreover, suppression of DHTKD1 leads to impaired mitochondrial biogenesis and increased reactive oxygen species (ROS), thus leading to retarded cell growth and increased cell apoptosis. These findings demonstrate that DHTKD1 contributes to mitochondrial biogenesis and function maintenance.

Background Circulating endothelial cells (CECs) are markers of vascular damage that have clinical relevance in many diseases, including acute myocardial infarction (AMI), and may be predictors of treatment responses. Herein, we investigated the diagnostic and prognostic value of CEC monitoring in AMI patients and a murine model. Methodology/Principal Findings CECs were defined as Hoechst 33342+/CD45−/CD31+/CD146+/CD133− in human blood samples and Hoechst 33342+/CD45−/CD31+/KDR+/CD117− in murine samples. To evaluate the validity and variability of our CEC detection system, peripheral blood samples of vascular endothelial growth factor-treated athymic nude mice and AMI patients were collected and subjected to intra-assay analysis. CEC detection by flow cytometry and real-time PCR were compared. Blood samples were obtained from 61 AMI patients, 45 healthy volunteers and 19 samples of the original AMI patients accepted one month treatment, via flow cytometry and expressed as a percentage of peripheral blood mononuclear cells. Results Our CEC detection method was validated and had limited variability. CEC concentrations were higher in AMI patients compared to healthy controls. One month post-treatment, CECs levels decreased significantly. Conclusions/Significance CEC levels may be useful as a diagnostic and prognostic biomarker in AMI patients.

Uveal melanoma (UM) is the most common primary intraocular malignancy and the leading potentially fatal primary intraocular disease in adults. Melanoma antigen recognized by T-cells (MART-1) has been studied extensively as a clinically important diagnostic marker for melanoma, however, its biological function remains unclear. In the present study, the UM cell line SP6.5, which showed a high level of MART-1 expression, was subjected to small interfering RNA-mediated silencing of MART-1. Silencing of MART-1 expression increased the migration ability of SP6.5 cells and down-regulated the expression of the metastasis suppressor NM23. Our results suggest that MART-1 is a candidate target for the development of therapeutic strategies for UM and in particular for the suppression of metastasis associated with this malignancy.

目的 探讨老年慢性心力衰竭患者血清同型半胱氨酸(homocysteine,HCY)的水平及其对心脏结构和功能的影响.方法 研究对象为2010年1月-2011年6月上海交通大学附属第一人民医院老年科住院患者,年龄70～94岁,平均(83.5+5.5)岁.包括慢性心力衰竭患者40例,非心力衰竭患者40例,心功能分级采用NYHA分级.血清HCY的水平采用酶联免疫法测定.采用超声心动图仪测定左室后壁厚度、室间隔厚度、左室舒张末内径和左室射血分数.结果 老年慢性心力衰竭组血清HCY高于非心力衰竭组(P＜0.05),而且慢性心力衰竭组患者心力衰竭程度随HCY的水平升高而加重(P=0.03).舒张末期内径与HCY呈正相关(r=0.394,P＜0.05),而左室射血分数与HCY水平呈负相关(r=-0.53,P＜0.01).结论 老年慢性心力衰竭患者组血清HCY高于非心力衰竭组.血清HCY是慢性心力衰竭的独立危险因素.

STUDY QUESTION:What are the implications of multiple alterations of the forkhead box L2 (FOXL2) gene in blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) patients?SUMMARY ANSWER:A multi-mutation of FOXL2, consisting of the expansion of the polyalanine tract from 14 to 24 residues (FOXL2-Ala24), an novel Y186C substitution from c.557A>G, and a synonymous variant (c.505G>A), had a cumulative effect on ovarian phenotypes in BPES patients.WHAT IS KNOWN ALREADY:Mutations in FOXL2, a gene encoding a forkhead transcription factor cause BPES. Overall, the expansion of the polyalanine tract of FOXL2 from 14 to 24 residues (FOXL2-Ala24) accounts for 30% of intragenic mutations.STUDY DESIGN, SIZE, DURATION:In this study, patients from seven BPES families and six sporadic cases were included.PARTICIPANTS/MATERIALS, SETTING, METHODS:We conducted an extensive clinical, hormonal and functional study in 20 patients carrying the expansion of the polyalanine tract of FOXL2 associated with BPES. A multi-mutation of FOXL2 was detected in one BPES family that showed more severe BPES symptoms. Subcellular localization and transactivation studies were performed for the constructs of FOXL2-Ala24, Y186C and FOXL2-Ala24-Y186C.MAIN RESULTS:We described the first multi-mutation of FOXL2 (c. [672_701dup30; 557A>G]) that leads to the polyalanine expansion of +10 residues (FOXL2-Ala24) combined with an Y186C substitution and a synonymous variant in a Chinese BPES family. This multi-mutation genotype was associated with more serious BPES clinical manifestations and the development of esotropia in the right eye. In in vitro studies, the multi-mutation affected the function of FOKL2 on the StAR promoter and DK3, and induced more aggressive aggregation and mislocalization of FOXL2 protein. The synonymous variant, while not affecting amino acid coding, causes a change in the RNA stem-loop structure.LIMITATIONS, REASONS FOR CAUTION:The multi-mutation of FOXL2 was detected in one BPES family and it needs to be validated further by more BPES subjects.WIDER IMPLICATIONS OF THE FINDINGS:The results of our study contribute new insights into the research field of BPES caused by the multi-mutation of FOXL2.STUDY FUNDING/COMPETING INTERESTS:This study was supported by Shanghai Leading Academic Discipline Project (Grant number S30205) and Shanghai Jiao Tong University School of Medicine Doctor Innovation Fund (Grant number 201131). The authors have no competing interests to declare.

Notch signaling pathway plays an important role in tumorigenesis by maintaining the activity of self-renewal of cancer stem cells, and therefore, it is hypothesized that interference of Notch signaling may inhibit tumor formation and progression. H101 is a recombinant oncolytic adenovirus that is cytolytic in cells lacking intact p53, but it is unable to eradicate caner stem cells. In this study, we tested a new strategy of tumor gene therapy by combining a Notch1-siRNA with H101 oncolytic adenovirus. In HeLa-S3 tumor cells, the combined therapy blocked the Notch pathway and induced apoptosis in tumors that are p53-inactive. In nude mice bearing xenograft tumors derived from HeLa-S3 cells, the combination of H101/Notch1-siRNA therapies inhibited tumor growth. Moreover, Notch1-siRNA increased Hexon gene expression at both the transcriptional and the translational levels, and promoted H101 replication in tumors, thereby enhancing the oncolytic activity of H101. These data demonstrate the feasibility to combine H101 p53-targted oncolysis and anti-Notch siRNA activities as a novel anti-cancer therapy.

Connexins play important roles in many physiological and pathological processes. Although connexins are considered as tumor suppressors in several types of cancer, their roles in pancreatic cancer are unknown. In this study, we found that connexin 43 (Cx43) caused apparent apoptosis and growth inhibition in pancreatic cancer cells. The tumor-suppressive role of Cx43 was independent of the canonical gap junction pathway. In the context of apoptosis, Cx43 translocated to the mitochondria, where it interacted with Bax to initiate the mitochondrial apoptotic pathway. Following further examination of the Cx43 protein, we found that the 241-382aa region of Cx43 was required for interaction with Bax. Furthermore, this region was responsible for permeabilizing mitochondrial membrane potential. The results from the present study elucidate a novel mechanism of the Cx43-mediated regulation of apoptosis in pancreatic cancer.

Vitamin K2 (VK2) exerts cell growth inhibitory effects in various human cancer cells such as SMMC-7721 hepatocellular carcinoma (HCC) cells. BCL-2 is an antiapoptotic protein that is frequently overexpressed in numerous tumors. Modulation of multiple antiapoptotic signaling pathways involving BCL-2, which are related to growth factor-stimulated signal transduction in cell survival, is essential for enhancement of the cytotoxic effect of anticancer drugs. In this study, we tested a new strategy of gene therapy by combining BCL-2 siRNA with VK2. In SMMC-7721 HCC cells, the combined treatment significantly enhanced cytotoxicity compared with treatment with either VK2 or siBCL-2 alone. We found that combined treatment induced a significantly different level of G2 stage inhibition. Furthermore, the p53 protein was overexpressed 24 h subsequent to combination treatment, and p21 was clearly increased at 36 h as a consequence of the increased p53 activity. In conclusion, these data suggest that the antitumor effect of VK2 may be improved by silencing BCL-2 expression in SMMC-7721 HCC cells and provides support for the combined use of VK2 and siBCL-2 as a promising approach in cancer gene therapy.

目的 探讨老年代谢综合征(MS)患者中血清游离脂肪酸(FFA)水平与胰岛素抵抗(IR)的相关性.方法 选取2009年1月至2010年6月上海交通大学附属第一人民医院老年科伟院患者96例,年龄≥65岁.采用2005年国际糖尿病联盟提出的标准,分为MS组38例及非MS组58例,分别检测血清FFA、胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、空腹血糖(FBG)、空腹胰岛素(FINS),用稳态模式(HOMA Model)公式计算HOMA-IR评估胰岛素抵抗.分别测各组体重、腰围、身高,计算体质指数(BMI).结果 1、MS组的身高、体重、腰围、BMI、FBG、FINS及HOMA-IR、TC、HDL-C、TG水平均高于非MS组(P<0.01),MS组的FFA水平高于非MS组(P<0.01);2、直线相关分析显示在MS组中FFA水平与FBG、FINS、HOMA-IR呈正相关(P<0.05);3、多元逐步回归分析结果显示FFA与HOMA-IR独寺相关(P<0.05).结论 老年MS患者FFA升高,FFA水平与IR存在相关性,FFA是老年MS患者IR的独立危险因素.

Oncolytic viruses have been developed as a new approach for the treatment of cancer.Oncolytic viruses in combination with radiotherapy may have an important impact for the treatment of metastatic cancer after the approval of oncolytic adenovirus for clinical tumor treatment.In this review,the killing effects on tumor cells of oncolytic viruses in combination with radiotherapy are discussed.

Uveal melanoma (UM) is one of the most common primary intraocular malignant tumors in adults. Melanoma antigen recognized by T cell-1 (Mart-1), one of the melanosome-specific proteins, has been widely studied as a marker recognized by cytotoxicity T lymphocytes. Mart-1 is considered to play a critical role in the immunotherapy for melanoma. Additionally, as a biomarker, Mart-1 is often used with other tumor-associated antigens for antidiastole in cutaneous melanoma (CM), uveal melanoma (UM) and nevus. In this study, the differential expression of Mart-1 was investigated in four human UM cells (SP6.5, VUP, OCM-1 and OM431) on three levels of analysis: messenger ribonucleic acid (mRNA), protein and, eventually, morphology. The results revealed that SP6.5 cells had high Mart-1 protein expression while VUP cells had almost none. OCM-1 and OM431 cells produced less Mart-1 than SP6.5 cells according to Western blot analysis, although OM431 cells had the highest expression of Mart-1 mRNA according to real-time PCR. The results indicate the potential use of Mart-1 in the development of therapy for UM.

There is a growing evidence that regucalcin (RGN) plays a multifunctional role in liver cancer cells. Previous reports showed that the presence of phorbol 12-myristate 13-acetate (PMA) caused a significant increase in RGN mRNA expression and promoter activity in rat hepatoma cells. In this study, we confirmed that human RGN is also up-regulated by PMA treatment independent of translation, and we identified the mechanism by which PMA up-regulates the expression of human RGN via driving SP1 away from a SP1 motif located within -188/180 of the promoter in HepG2 cells. Overexpression of SP1 dramatically reduces PMA-induced up-regulation of both internal expression of mRNA and promoter activity, whereas knockdown of SP1 has the opposite effect. Therefore, the present study delineates the fundamental elements in the promoter which will be helpful in the future studies on the regulation of RGN expression in liver cancer.