The Targeted Pro-apoptotic Effect of scFv-mediated Delivery of Caspase-6 on HER2-positive Osteosarcoma Cells

Objective To investigate the pro-apoptotic effect of scFv-mediated delivery of caspase-6 fusion protein on osteosarcoma E 10 cells. Methods The expression of human epidermal growth factor receptor-2 （Her-2） in osteosarcoma cell line E10 was examined by flow cytometry analysis. ScFv-mediated delivery of caspase-6 was generated by sequential fusion of the genes of a signal peptide, a single-chain Her-2 antibody （e23sFv）, a PEA translocation domain （PEA aa253-264）, and an recombinant caspase-6. The pCMV-immunocaspase-6 was produced by cloning above mentioned fusion genes into pCMV plasmid, and then pCMV-immunocaspase-6 plasmid mixed cationic liposome was transfected into E10 cells. The expression of fusion gene on transfected E10 cells was detected by immunocytochemical staining, The pro-apoptotic effects of fusion gene expression on morphology and growth status of E10 cells were observed by trypan blue cell staining, electron microscopy and MTT assay. Results The fusion protein was detected in the cytoplasm of the transfected E10 cells. These cells presented typical characteristics of apoptosis as detected by electron microscopy. MTT assay revealed that the proliferation of transfected E10 cells was suppressed than that of non- or mock-transfected cells （P〈0.01）. Conclusion ScFv-mediated delivery of caspase-6 fusion protein can induce apoptosis of Her-2 positive osteosarcoma E10 cells, suggesting the potential of this strategy for the treatment of human high-grade osteosarcoma.     

超临界CO2萃取和水蒸气蒸馏法提取苍耳子挥发油的比较研究

目的比较两种不同方法提取的苍耳子挥发油化学成分。方法采用超临界CO2萃取法、水蒸气蒸馏法进行提取。气相色谱-质谱法进行成分分析。结果共鉴定出74个成分,超临界CO2萃取法提取的挥发油被鉴定的成分有69个,水蒸气蒸馏法提取的挥发油被鉴定的成分有63个,两者共有成分58个。结论两种方法得到的挥发油主要成分大部分一致,但相同成分在两种提取方法中含量也有不同,SEE—CO2方法不能完全替代SD法。     

Effect of Shenfu on the Expression of Bcl-2 and Bax of Intestinal Mucosal Epithelial Cells after Ischemic Reperfusion in Rats

Objective To study the effect of Shenfu on the expression of bax and bcl-2 in small intestinal mucosal epithelial cells after ischemic reperfusion injury and explore the effect of Shenfu on small intestinal mucosal epithelial cells apoptosis. Methods 36 SD rats were randomized into three groups, each consisting of 12 rats： Sham group （S group）, Ischemic reperfusion group （IR group）, Shenfu group （SF group）, Ischemic reperfusion models were made by ligated the superior menseneric artery for 1 hour followed by 2 hrs reperfusion. Histological mucosal damage in each group was graded according to Chiu＇s score. Immunohistochemistry detected the expression of bax and bcl-2, and obtained the optical density （OD） value using a color image pattern analysis system, and then calculated the ratio of bcl-2/bax. TUNEL method measured apoptotic intestinal mucosal epithelial cells, and calculated the apoptotic index. Results There were edema and epithelial impairment in part of villus and the integral was higher in SF group than S group （P〈0.05）, but lower than IR group （P〈0.01）. Compared IR group with S group and SF group, the average OD value of bcl-2 and bax had significant statistic difference （P〈0.01）, and the average OD value of bcl-2 in SF group was higher than in S group （P〈0.05）, meanwhile bcl-2/bax was significantly lower in IR group than in S group and SF group （P〈0.01）, but that was higher in SF group than in S group （P〈0.05）. Apoptotic index in IR group was significantly higher than that in other two groups （P〈0.01）, and that in SF group was higher than in S group （P〈0.05）. Conclusion Shenfu can enhance the expression of bcl-2, decrease the expression of bax, meanwhile increase the ratio of bcl-2/bax, inhibit small intestinal mucosal epithetlial cells apoptosis, and protect small intestine mucosal epithetlial after ischemic reperfusion.     

MiR-503 regulates cisplatin resistance of human gastric cancer cell lines by targeting IGF1R and BCL2

Background Studies have shown that the drug resistance of gastric cancer cells can be modulated by abnormal expression of microRNAs （miRNAs）.We investigated the role of miR-503 in the development of cisplatin resistance in human gastric cancer cell lines.Methods MiR-503 expression was measured by quantitative real-time PCR.MTT （3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide） and clonogenic assays were used to examine changes in cell viability and the drug resistance phenotype of cancer cells associated with upregulation or downregulation of the miRNA.A dual-luciferase activity assay was used to verify target genes of miR-503.Immunohistochemistry,Western blotting analysis,and a flow cytometric apoptosis assay were used to elucidate the mechanism by which miR-503 modulates drug resistance in cancer cells.Results MiR-503 was significantly downregulated in gastric cancer tissues and several gastric cancer cell lines.Additionally,downregulation of miR-503 in the cisplatin （DDP）-resistant gastric cancer cell line SGC7901/DDP was concurrent with the upregulation of insulin-like growth factor-1 receptor （IGF1R） and B-cell lymphoma 2 （BCL2） expression compared with the parental SGC7901 cell line.An in vitro drug sensitivity assay showed that overexpression of miR-503 sensitized SGC7901/DDP cells to cisplatin.The luciferase activity of reporters driven by IGF1R and BCL2 3＇-untranslated regions in SGC7901/DDP cells suggested that IGF1R and BCL2 were both direct target genes of miR-503.Enforced miR-503 expression in SGC7901/DDP cells reduced expression of the target proteins,inhibited proliferation,and sensitized the cells to DDP-induced apoptosis.Conclusion Our findings suggest that hsa-miR-503 modulates cisplatin resistance of human gastric cancer cells at least in part by targeting IGF1R and BCL2.     

益智健脑颗粒联合针灸对阿尔茨海默病大鼠学习记忆的影响

目的观察益智健脑颗粒联合针灸对阿尔茨海默病（Alzheimers disease,AD）大鼠学习记忆的影响。方法将大鼠随机分为假手术组（A组）、模型组（B组）、针灸组（C组）、益智＋针灸组（D组）各10只,B、C、D 3组分别以海马CA1区注射β淀粉样蛋白25-35（Aβ25-35）造模,A组注射等量的双蒸水,各组分别治疗20 d后行Morris水迷宫试验,观察大鼠学习记忆能力变化。结果B组较A组的平均潜伏期明显延长,差异具有统计学意义（P〈0.05）;与B组比较,C组、D组的平均潜伏期明显缩短,过台次数增多,差异具有统计学意义（P〈0.05,P〈0.01）;与C组比较,D组的潜伏期缩短,过台次数增多,差异具有统计学意义（P〈0.05）。结论益智健脑颗粒联合针灸能够提高Aβ25-35介导的AD模型大鼠的学习记忆能力。     

Interaction of sleep quality and sleep duration on glycemic control in patients with type 2 diabetes mellitus

Background Copious evidence from epidemiological and laboratory studies has revealed that sleep status is associated with glucose intolerance, insulin resistance, thus increasing the risk of developing type 2 diabetes. The aim of this study was to reveal the interaction of sleep quality and sleep quantity on glycemic control in patients with type 2 diabetes mellitus. Methods From May 2013 to May 2014, a total of 551 type 2 diabetes patients in Tianjin Metabolic Diseases Hospital were enrolled. Blood samples were taken to measure glycosylated hemoglobin （HbAlc）, and all the patients completed the Chinese version of the Pittsburgh Sleep Quality Index （PSQI） questionnaire to evaluate their sleep status. ＂Good sleep quality＂ was defined as PQSI 〈5, ＂average sleep quality＂ was defined as PQSI 6-8, and ＂poor sleep quality＂ was defined as PQSI 〉8. Poor glycemic control was defined as HbAlc 〉7%. Sleep quantity was categorized as 〈6, 6-8, and 〉8 hours/ night. Short sleep time was defined as sleep duration 〈6 hours/night. Results In the poor glycemic control group, the rate of patients who had insufficient sleep was much higher than that in the other group （X2=11.16, P=0.037）. The rate of poor sleep quality in poor glycemic control group was much greater than that in the average control group （X2=9.79, P=-0.007）. After adjusted by gender, age, body mass index, and disease duration, the adjusted PSQI score＇s OR was 1.048 （95% CI 1.007-1.092, P=0.023） for HbAlc level. The sleep duration＇s OR was 0.464 （95% CI 0.236-0.912, P=0.026） for HbAlc level. One-way analysis of variance showed that the poor sleep quality group had the highest homeostasis model assessment-insulin resistance （P 〈0.01）. Conclusions Inadequate sleep, in both quality and quantity, should be regarded as a plausible risk factor for glycemic control in type 2 diabetes. Poor sleep might bring much more serious insulin resistance and could be the reason for bad glycemic control. A good night＇s sleep sho     

Prevalence of stroke and metabolic disorders in the middle-aged and elderly Chinese with type 2 diabetes

Background Stroke is now the most prevalent and debilitating disease affecting diabetic population in China. The study aimed to investigate the prevalence of stroke and metabolic disorders in the middle-aged and elderly Chinese with type 2 diabetes. Methods A total of 4 629 subjects with type 2 diabetes （males： 1 917; females： 2 712） aged ≥ 40 years from Shijingshan district, Beijing, China from November 2011 to August 2012 were included in the study. Data on demographic information, lifestyle, history of diabetes mellitus, stroke, coronary heart disease, hypertension, and dyslipidemia were collected. The oral glucose tolerance test or a standard meal test was performed. Non-fatal stroke was reported by the subjects. The 2-tailed test was used, and P 〈0.05 was regarded as statistically significant. Results Prevalence of stroke in the subjects with type 2 diabetes was 5.5%. The prevalence of smoking, overweight or obesity, hypertension, and dyslipidemia was 41.0%, 65.8%, 67.4%, and 52.0% in males, and 2.2%, 65.5%, 69.5%, and 57.6% in females. Multivariate Logistic regression analysis showed that increased age, hypertension, diabetic duration, and overweight or obesity were positively correlated with stroke in the population with type 2 diabetes, whereas high- density lipoprotein cholesterol level was negatively correlated with stroke. After adjustment for age and gender, the odds ratio values of stroke in subjects having 1,2 or ≥3 of 4 risk factors, including smoking, overweight or obesity, hypertension and dyslipidemia, were 2.302 （95% CI： 0.789-6.712）, 4.089 （95% CI： 1.470-11.373）, 6.023 （95% CI： 2.176-16.666）, compared with subjects without any of the above 4 risk factors. Conclusions The prevalence of stroke was higher in middle-aged and eldedy Chinese with type 2 diabetes than that in the general population. With the aggregation of risk factors, the prevalence of stroke increased.     

Prognostic significance of γ-H2AX in laryngeal squamous cell carcinoma after surgery

Background The clinical significance of Y-H2AX in laryngeal squamous cell carcinoma （LSCC） has not yet been established. This study was performed to assess the expression of nuclear y-H2AX in benign and malignant laryngeal lesions and to assess its clinicopathological significance. Methods A total of 70 LSCC tumor-normal tissue paired samples were evaluated for y-H2AX expression using immunohistochemical staining. Their expression was correlated with different clinicopathological parameters. Results Nuclear Y-H2AX expression was frequently detected in LSCC tissues （P 〈0.001）. High nuclear Y-H2AX levels were not associated with any clinicopathological characteristics of LSCC （P 〉0.05）. Univariate Kaplan-Meier analysis showed that positive nuclear Y-H2AX expression was associated with a decreased overall survival （P=0.017）. Multivariate Cox regression analysis showed that nuclear Y-H2AX expression was an independent risk factor for overall survival. Conclusion The expression of nuclear Y-H2AX might be closely related to the prognosis of LSCC. Chin Med J 2014;127 （14）： 2664-2667     

2型糖尿病并发急性心肌梗死的临床特点分析

我国是2型糖尿病的高发国家,随着生活水平的提高,饮食结构的改变,日趋紧张的生活节奏以及诸多不良的生活方式,我国糖尿病、冠心病的发病率增长迅速,糖尿病患者发生心肌梗死的发生率也明显增多。总结归纳其特点,以指导临床。现将我院近年来糖尿病合并急性心梗的患者资料进行总结归纳,临床分析。1资料与方法1.1病例选择选择自2004年1月至2007年12月我院急性心肌梗死住院病例398例,分成两组,A组:T2DM发生AMl98例,其中男63例、女35例;B组:NT2DM发生AMI300例,男180例,女120例,两组年龄无差异。T2DM患者按1997年美国糖尿病协会分类标准为T2DM,AMI患者的论断均有18导联心电图,血清心肌酶改变的支持。1.2方法两组患者入院至入院后2周,第1天、第2天、第3天每日上午8∶00、下午2∶00做心电图及心肌酶学、血糖,以后隔日做心电图,心肌酶学、血糖、动态观察心功能状态。1.3分析处理采用对比分析方法,资料中组间率的显著性检验使用χ2检验。2结果表1两组间比较(例)项目A例组数(n=9%8)B例组数(n=30%0)P无痛性心梗4545.928327.67<0.05非ST段抬高心梗4343....     

Norcantharidin inhibits tumor growth and vasculogenic mimicry of human gallbladder carcinomas by suppression of the PI3-K/MMPs/Ln-5T2 signaling pathway

Background：Vasculogenic mimicry （VM） is a novel tumor blood supply in some highly aggressive malignant tumors. Recently,we reported VM existed in gallbladder carcinomas （GBCs） and the formation of the special passage through the activation of the PI3K/ MMPs/Ln-5γ2 signaling pathway. GBC is a highly aggressive malignant tumor with disappointing treatments and a poor prognosis. Norcantharidin （NCTD） has shown to have multiple antitumor activities against GBCs, etc; however the exact mechanism is not thoroughly elucidated. In this study, we firstly investigated the anti-VM activity of NCTD as a VM inhibitor for GBCs and its underlying mechanisms. Methods ： In vitro and in vivo experiments to determine the effects of NCTD on proliferation, invasion, migration, VM formation, hemodynamic and tumor growth of GBC-SD cells and xenografts were respectively done by proliferation, invasion,migration assays,HE staining and CD31-PAS double staining, optic/electron microscopy, tumor assay, and dynamic micro- MRA. Further, immunohistochemistry, immunofluorescence, Western blotting and RT-PCR were respectively used to examine expression of VM signaling-related markers PI3-K,MMP-2 ,MT1-MMP and Ln-5γ2 in GBC-SD cells and xenografts in vitro and in vivo. Results： After treatment with NCTD, proliferation, invasion, migration of GBC-SD cells were inhibited; GBC-SD ceils and xenografts were unable to form VM-like structures; tumor center-VM region of the xenografts exhibited a decreased signal in intensity; then cell or xenograft growth was inhibited. Whereas all of untreated GBC-SD cells and xenografts formed VM-like structures with the same conditions; the xenograft center-VM region exhibited a gradually increased signal; and facilitated cell or xenograft growth （Figure 1-6 ）. Furthermore, expression of MMP-2 and MT1-MMP products from sections/supemates of 3-D matrices and the xenografts, and expression of PI3-K, MMP-2, MM1-MMP and Ln-5γ2 proteins/mRNAs of the xenografts were all decreased in NCTD or TIMP-2 g     

Understanding the standard of care in the treatment of type 2 diabetes in China: results from a national survey

Background Given the pace with which standards of care have changed, timely assessment of their impact on routine clinical practice and patient outcomes is needed. In coordination with the Chinese Diabetes Society （CDS）, we developed a quantitative survey to explore the implementation of standards of care for type 2 diabetes （T2D） in China. Methods A national online survey of physicians involved in the management of T2D in China was conducted over a 4-week period in 2012. Completed responses were obtained from 1 028 physicians. Participants responded to 52 questions designed to capture information relating to their demographic and clinical practice profiles. The questionnaire was divided into three sections： basic information, diagnosis practices and screening methods on main complications, and treatment and control practices. The questionnaire was developed in conjunction with the CDS. Results Overall, 83% of surveyed physicians were at least ＂aware＂ of the CDS guidelines on standards of care for T2D. Level of awareness was directly related to hospital grade, specialty, geographic location, professional rank and participation in CDS training. The 2-hour oral glucose tolerance test was reported as the most ever-used approach across all three hospital grades and physician specialties, with a usage rate of 97%. Respondents selected their choice of primary treatment for newly diagnosed T2D patients. Just over half （52%） indicated the use of oral anti-diabetic drugs （OAD） monotherapy, in line with CDS recommendations. However, OAD use varied considerably between different regions and city tiers. Despite hemoglobin Alc being defined as the gold standard for glucose control, it was not universally measured, with more physicians indicating routine use of glucose before fasting and glucose non-fasting. Conclusion The standards-of-care analysis has provided important insights into the current management of T2D among physicians in China across different geographical regions, hospital grades, specialti     

WHO西太区与“世界中医药学会联合会”中医名词术语国际标准比较研究：精、神、气、血、津、液部分

精、神、气、血、津、液是中医理论中6个非常重要的概念，有关术语在《WHO西太区传统医学国际标准名词术语》中一共收录了59条，“世界中医药学会联合会”（以下简称世中联）《中医基本名词术语中英对照国际标准》中收录了58条。血、津、液的内涵较为具体，有一定的物质基础，理解并不困难。翻译上虽有差异，但亦不难统一。精、神、     

Expression and Significance of Cox-2 and Livin in Oral Squamous Cell Carcinoma with Tissue Microarray

Objective To detect the expression of Cox-2 and livin in oral squamous cell carcinoma and precancerous lesion with tissue microarray, and discuss their significance and relationship in the occurrence and development of oral squamous cell carcinoma. Methods Immunohistochemical （IHC） staining and tissue microarray technique were used to detect the expression of Cox-2 and livin in noma! oral mucous membrane, precancerous lesion and oral squamous cell carcinomas. Results The expression of Cox-2 was negative in normal oral mucous membrane, and positive in precancerous lesion （81.6%） and squamous carcinoma （85.2%）; while the expression of livin was negative or weakly positive in normal oral mucous membrane, and positive in precancerous lesion （89.8%） and squamous carcinoma （100%）. The positive expression of Cox-2 and livin were both closely related to pathological classifications of oral squamous cell carcinomas. But there was no correlation between them. Conclusion Cox-2 and livin have close relationship with the occurrence and development of oral squamous cell carcinoma, but no correlation with the expression.     

糖络方联合弥可保治疗2型糖尿病周围神经病变的临床观察

目的观察益气养阴活血通络之糖络方联合弥可保治疗2型糖尿病周围神经病变的疗效。方法将65例患者随机分成治疗组（中西药组）和对照组（西药组）,均在西药降血糖基本达标的同时,对照组单纯口服弥可保,治疗组口服弥可保的基础上加用糖络方内服,治疗8周。结果总有效率及治疗后证候积分比较,治疗组优于对照组,差异有统计学意义（P〈0．05）;治疗后肌电图比较,治疗组优于对照组,差异有统计学意义（P〈0．01）。结论益气养阴活血通络之糖络方联合弥可保治疗2型糖尿病周围神经病变疗效优于单用弥可保,是治疗该病的有效方剂之一。     

An update of molecular pathogenesis and diagnosis of myeloproliferative disorders in the JAK2 era

Myeloproliferative disorders （MPD） are clonal haematopoietic stem cell disorders characterized by proliferation of one or more myeloid cell lineages in the bone marrow and increased numbers of mature and immature cells in the peripheral blood. MPDs are classified into five categories： polycythemia vera （PV）, essential thrombocythaemia （ET）, idiopathic myelofibrosis （IMF）, chronic myelogenous leukaemia （CML） and atypical MPD. The atypical MPD includes chronic myelomonocytic leukaemia, juvenile myelomonocytic leukaemia, chronic neutrophilic leukaemia, chronic eosinophilic leukaemia, chronic basophilic leukaemia, hypereosinophilic syndrome, systemic mastocytosis, atypical CML and unclassifiable cases.1-3 Apart from the Philadelphia chromosome and BCR-ABL fusion gene as the characteristic genetic abnormality of CML, the molecular pathogenesis of most MPDs such as PV, ET and IMF has not been described. Nonspecific cytogenetic abnormalities were found at diagnosis in numerous patients including deletions of the long arms of chromosome 20 and chromosome 13,     

郭振球教授高血压病辨治特色

郭振球教授是湖南中医药大学教授,从事临床、科研、教学工作六十一年,积累了丰富的临床经验,在学术上治学严谨。学验俱丰。1986年人选我国首批中医学博士研究生导师．1990年被评为我国首批全国继承老中医学术经验指导老师,开创了微观证治学,系世界传统医学诊断学学科奠基人。     

Effect of behavior training on learning,memory and the expression of NR2B,GluR1 in hippocampus of rats offspring with fetal growth restriction

Objective： To study effects of behavior training on learning, memory and the expression of NR2B, GluR1 in hippocampus of rat＇ s offspring with fetal growth restriction（FGR）. Methods： The rat model of FGR was established by passive smoking method. The rats offspring were divided into the FGR group and the control group, then randomly divided into the trained and untrained group, respectively. Morris water maze test was proceeded on postnatal month（PM2/4） as a behavior training method, then the learning-memory of rats was detected through dark-avoidance and step-down tests. The expressions of NR2B and GluR1 subunits in hippocampal CA1 and CA3 areas were detected by immunohistochemical method. Results： In the dark-avoidance and step-down tests, the performance record of rats with FGR was worse than that of control rats, and the behavior-trained rats was better than the untrained rats, when the FGR model and training factors were analyzed singly. The model factor and training factor had significant interaction（P 〈 0.05）. The expressions of NR2B and GluR1 subunits in hippocampal CA1 and CA3 areas of rats with FGR reduced. In contrast, the expressions of GluR1 and NR2B subunits in CA1 area of behavior-trained rats increased, when the FGR model and training factors were analyzed singly. Conclusion： These findings suggested that the effect of behavior training on the expressions of NR2B and GluR1 subunits in CA1 area should be the mechanistic basis for the training-induced improvement in learning-memory abilities.     

Comparative study of mutation spectrums of MT-RNR1 m.1555A>G, GJB2, and SLC26A4 between familial and sporadic patients with nonsyndromic sensorineural hearing loss in Chinese Han

Background The mutation frequencies of three common deafness genes （MT-RNR1 m.1555A〉G,GJB2,and SLC26A4） among patients with nonsyndromic sensorineural hearing loss （NSHL） were different in previous studies.Inconsistent selection criteria for recruiting patients could have led to differences in estimating the frequencies of genetic mutations thus resulting in different mutation frequencies among these studies.The aim of this study was to reveal the differences in the mutation spectrums of the three common genes between familial and sporadic Chinese Han patients.Methods Totally,301 familial probands and 703 sporadic patients with NSHL were enrolled in this study.Three genes,MT-RNR1 m.1555A〉G,GJB2,and SLC26A4,were screened for mutation in our study cohort.A X2 test was performed to compare the mutation frequencies between the two groups.Results The study showed that the disease-causing mutation frequencies of MT-RNR1 m.1555A〉G,GJB2,and SLC26A4 were 12.29%,14.62%,and 18.27% in familial probands and 3.56%,18.63%,and 18.92% in sporadic patients,respectively.The mutation frequency of MT-RNR1 m.1555A〉G in familial probands was significantly higher than in sporadic patients （X2 test,P=0.000）,while there were no significant differences in the mutation frequencies of GJB2 and SLC26A4 between the familial and sporadic groups （X2 test,P 〉0.05）.Conclusions It is necessary to reveal the differences in gene mutation frequencies between patients of different sources or characteristics by comparative studies in order to avoid selection bias.The mutations of GJB2,SLC26A4,and MTRNR1 m.1555A〉G are the most important etiological factors in Chinese Han patients,among which SLC26A4 might be the most frequent.