Objective To assess the effect of suppression of insulin-like growth factor-1 receptor (IGF1R)in HO8910PM cell line by small interference RNA(siRNA).Methods Transfection of siRNA using liporectamine 2000 was conducted to silence IGF1R gene expression,the expression levels of IGF1R mRNA and protein were evaluated,and the effects on the cell cycles at 48 hours of transfection were assessed by real-time PCR,western blot and flow cytometry(FCM)assay respectively.The cell growth was detected by cell counting kit-8(CCK-8)at 24,48,72,96 hours of transfection.After 24 hours of transfection,the cells were cuhured with difierent concentrations of cisplatin(DDP)for 24 hours,the cell growth inhibition rate Was evaluated by CCK-8.Following incubation with 10μg/ml DDP for 24 hours after 24 hours of transfection,the apoptosis cells and the protein expression level of apoptosis-related gene,B cell leukemia/lymphoma 2(Bcl-2),were identified by FCM and western blot respectively.Resuits (1)Expression levels of IGF1R mRNA and protein were markedly decreased respectively at 48 hours of transfection IGF1R siRNA.(2)Suppression of IGF1R accompanied the reduction of cell growth at 48,72,96 hours of transfection with IGF1R siRNA,absorbance were 1.71±0.13,2.32±0.23,2.79±0.28 respectively (P＜0.01).(3)IGF1R siRNA induces arrest of G2 phase,the G2 phase rate of cells were 24.37%(P＜0.05).(4)Following treatment with 2.5,5,10,20μg/ml DDP for 24 hours after 24 hours of transfection,the cell growth inhibition rates were(25.94±0.08)%,(40.25±0.05)%,(59.48±0.03)%and(74.18±0.08)%respectively(P＜0.01).(5)Treatment with 10μg/ml DDP for 24 hours after 24 hours of transfection,induces 17.95%of cells apoptosis(P＜0.05),and decreases Bcl-2 protein level.Conclusion RNA interference of IGF1R gene induces the IGF1R silence in HO8910PM cell line significantly,inhibits cell growth in vitro, arrests the G2 phase, and enhances the chemosensitization to DDP.

Adult ; Aged ; Carcinoma, Squamous Cell ; surgery ; Esophagus ; surgery ; Female ; Humans ; Laryngeal Neoplasms ; surgery ; Laryngectomy ; Male ; Middle Aged ; Phonation ; Trachea ; surgery

To prepare the RNAse-resistant virus particles containing the partial gene fragments of avian influenza virus H5N1 for use as RNA standard and control in RNA virus detection, the genes coding the coat protein and maturase of E.coli bacteriophage MS2 were amplified by PCR and then cloned into prokaryotic expression vector pET32a to construct the intermediate vector pET32a-MS2. In addition, the gene sequences coding hemagglutinin (HA), neuraminidase(NA) and M protein of the H5N1 virus were also cloned separately to the down-stream of plasmid pET32a-MS2, thus constructing the prokaryotic expression vectors pET32a-NS2-HA, pET32a-MS2-NA and pET32a-MS2-M. These recombinant plasmids were then transformed separately to E.coli BL21(DE3) with induction by IPTG. to express the virus-like particles. The virus-like particles observed under electron microscopy were identified by RT-PCR ,while their stability was confirmed by real-time RT-PCR. In this way, the virus-like particles were successively constructed and identified through PCR amplification, enzymolysis identification and sequencing analysis. These virus-like particles observed under electron microscopy appeared to be circular in shape with a diameter of about 50 nm. Their stability was proved to be rather good. From these observations, it is apparent that these virus-like particles can be used as RNA standard and quality control in the detection of avian influenza virus H5N1.

Objective To investigate the potential relationship between initial 131Ⅰ uptake by lung metastases from differentiated thyroid cancer (DTC) and treatment outcomes. Methods From 1997 to 2009, 41 patients with DTC lung metastases were treated in the authors' department. 131Ⅰ whole body scan (WBS), serum Tg levels and other imaging results were analyzed to evaluate the therapeutic effects. Complete response (CR) or partial response (PR) was considered to be effective. The x2 test and correlation analysis were performed using SPSS 11.5 software package. Results 131 Ⅰ treatment was effective in 63% (26/41) patients with DTC lung metastases, CR in 8 patients and PR in 18 patients. In other 37% ( 15/41 ) patients, 131Ⅰ treatment was ineffective, including one case died of distant metastases. Patients with initial presence of 131Ⅰ lung uptake had higher effective rate than those with 131Ⅰ lung uptake during the second or later 131Ⅰ treatment (76% (22/29)vs33% (4/12),x2 =4.911, P=0.027). Also, significantly higher effective rate was found in patients with lung metastases alone than those with extra-pulmonary metastases (75% (24/32) vs 22% (2/9), x2 = 6. 312, P =0.012). However, the effective rate in patients with diffuse metastases was not significantly different from that in patients with focal metastases (67% (12/18)vs 61% ( 14/23), x2 =0. 146, P=0.702). The positive rate of initial 131Ⅰ uptake by lung metastases was higher in patients with total thyroidectomy than those with partial thyroidectomy (83% (24/29) vs 42% (5/12) ). Those positive rates in patients with papilary DTC and patients with follicular DTC were 72% (23/32) and 6/9, respectively. The surgical mode was correlated with the initial 131Ⅰ uptake by lung metastases (r = 0.411, P ＜ 0.05), but no correlation was found between the histological type and the initial 131Ⅰ uptake by lung metastases ( r = 0. 047, P ＞ 0.05 ). Conclusion Initial uptake of 131 Ⅰ by lung metastases alone is a favorable prognostic factor for DTC patients treated by131Ⅰ, and total thyroidectomy may be beneficial for initial 131Ⅰ uptake by lung metastases.

Aged, 80 and over ; Heart Valve Prosthesis Implantation ; methods ; Humans ; Male ; Prosthesis Failure

Objective To explore the feasibility of evaluating the lung function by MSCT in emphysema.Methods The MSCT scan and pulmonary function tests(PFF)were respectively performed in 147 receptors within one week.They were randomly divided into 2 groups:group A(120 receptors), including normal,mild,moderate and severe abnormal pulmonary function based on the PFT,for comparing the correlation between pulmonary quantitative indexes of MSCT pulmonary function and PFT and settingup the primary grade criteria of abnormal pulmonary function in emphysema,group B(27 receptors)for evaluating the diagnostic accuracy in group A.The total lung was respectively scanned at the full inspiration and full expiration with MSCT.The pulmonary quantitative indexes of MSCT were measured with Siemens Pulmo pulmonary quantitative software.Results There was correlation between pulmonary quantitative indexes of MSCT and PFF.The Piex/in_(-910)showed best correlation with FEV_1%(r=-0.905,P

OBJECTIVE:To investigate the differences of the active components of different parts of rhubarb(the head,the body and the end part of the root)from genuine medicinal materials in Gansu province so as to provide theoretical basis for its quality control.METHODS:A comparative study of the fingerprints of different parts of rhubarb extract were conducted by HPLC,in which,the C18 was used as the chromatographic column under room temperature,the mobile phase consisted of menthol-0.1%H3PO4(70∶30)with a flow rate of 1.0ml/min,the column's temperature of room temperature and detection wavel_ ength of 280nm.RESULTS:Significant differences were noted in the HPLC fingerprints of different parts of rhubarb,there were significant differences in the contents of the active components such as emodin,chrysophanol,etc.,the contents decreased progressively from the head of root,the body part to the end part.CONCLUSION:In order to ensure the medical value and the quality of rhubarb medicinal material,different parts of which should be differentiated in the quality control.

OBJECTIVETo explore the location of telocytes in pulmonary tissues.

METHODSElectron microscopy, immunohistochemistry, primary cell culture and vital cell staining were used to identify the distribution of telocytes in mouse bronchial and pulmonary tissues.

RESULTSTelocytes were identified in the interstitial space between bronchial cartilage (cricoid) and smooth muscle by scanning and transmission electron microscope in mouse. By transmission electron micrscope and immunohistochemistry, telocytes were found in the interstitial spaces of lung parenchyma in connection with capillaries and bronchia. Telocytes expressed CD34, c-kit and vimetin by immunohistochemistry. After isolation, cultured telocytes demonstrated typical morphological feature, i.e. cells with telopode, which was seen as luminal structures with alternating thin and thick segments under electron microscope.

CONCLUSIONTelocytes are present in the interstitial space between cricoid cartilage and smooth muscle.

Animals ; Antigens, CD34 ; metabolism ; Bronchi ; cytology ; metabolism ; ultrastructure ; Cell Culture Techniques ; Immunohistochemistry ; Interstitial Cells of Cajal ; cytology ; metabolism ; ultrastructure ; Lung ; cytology ; metabolism ; ultrastructure ; Mice ; Mice, Inbred BALB C ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Proto-Oncogene Proteins c-kit ; metabolism ; Vimentin ; metabolism

In order to study the roles of integrin beta6 in Foot-and-Mouth Disease Virus infection, pig integrin beta6 was firstly molecularly cloned from RNA of the tongue and lung of recovered pig infected experimentally with foot-and-mouth-disease virus (FMDV), and was compared with the beta6 gene of other animals available in GenBank at nucleotide and amino acid leves. GeneBank association number of the beta6 gene is EF432729. Pig integrin beta6 gene (2367bp) encodes a polypeptide of 788 amino acids consisting of 9 potential N-linked glycosylation sites, 3 Glycosaminoglycan attachment sites, a cGMP-dependent protein kinase phosphorylation site, 10 Protein kinase C phosphorylation sites, 2 EGF-like domains and 2 cysteine-rich regions. Pig integrin beta6 subunit has a 26-residue putative signal peptide, a 681-residue ectodomain, a 29-residue transmembrane domain, and a 52-residue cytoplasmic domain. 11 mutant nucleotides were found in beta6 gene coding region and 9 amino acids were changed. The nucleotide sequence similarity of integrin beta6 gene between rheses monkey, mouse, Norway rat, dog, guinea pig, human, bovine, sheep is 79.5%, 84.9%, 85.4%, 85.2%, 88.7%, 90.1%, 91.9% and 91.9%, and the amino acid sequence similarity is 93.5%, 88.2%, 88.5%, 88.3%, 91.0%, 92.8%, 93.3% and 93.4% respectively. This study will lay a foundation for understanding the interactions of FMDV with receptors.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; Foot-and-Mouth Disease Virus ; pathogenicity ; Integrin beta Chains ; genetics ; metabolism ; Molecular Sequence Data ; Mutation ; Receptors, Virus ; genetics ; metabolism ; Sequence Analysis ; Swine ; genetics

BACKGROUNDTelocytes (TCs) are a novel type of interstitial cells, which have been recently described in a large variety of cavitary and noncavitary organs. TCs have small cell bodies, and remarkably thin, long, and moniliform prolongations called telopodes (Tps). Until now, TCs have been found in various loose connective tissues surrounding the arterioles, venules, and capillaries, but as a histological cellular component, whether TCs exist in large arteries remains unexplored.

METHODSTCs were identified by transmission electron microscope in the aortic arch of male C57BL/6 mice.

RESULTSTCs in aortic arch had small cell bodies (length: 6.06-13.02 μm; width: 1.05-4.25 μm) with characteristics of specific long (7.74-39.05 μm), thin, and moniliform Tps; TCs distributed in the whole connective tissue layer of tunica adventitia: TCs in the innermost layer of tunica adventitia, located at the juncture between media and adventitia, with their long axes oriented parallel to the outer elastic membrane; and TCs in outer layers of tunica adventitia, were embedded among transverse and longitudinal oriented collagen fibers, forming a highly complex three-dimensional meshwork. Moreover, desmosomes were observed, serving as pathways connecting neighboring Tps. In addition, vesicles shed from the surface of TCs into the extracellular matrix, participating in some biological processes.

CONCLUSIONSTCs in aorta arch are a newly recognized complement distinct from other interstitial cells in large arteries, such as fibroblasts. And further biologically functional correlations need to be elucidated.

Adventitia ; cytology ; Animals ; Aorta ; cytology ; Aorta, Thoracic ; cytology ; Cell Communication ; physiology ; Connective Tissue Cells ; cytology ; ultrastructure ; Male ; Mice ; Mice, Inbred C57BL ; Microscopy, Electron, Transmission