The surface and ultrastructure of the interphase and mitotic ECa 109 esophagealcancer epithelial cells were investigated by means of scanning and transmission electronmicroscopes.The electron microscopies revealed that cells grown in culture haddifferent external appearances during the different phases of cell cycle.At interphase,the cells were flattened and irregular in shapes,the surface were covered with microvilli.Centrosome,tonofilaments,mitochondria could be found in the cytoplasm.The largeand irregular shaped nucleus was located at the centre.It had one or more nucleoli.During the mitotic phase the cells were spherical and they became first cylindricaland then dumb-bell in shapes,Finally,they divided into daughter cells.There werespindle area and non-spindle area in the cell.The former was at the center of thecell and the latter occupied the outer part of the cell.In the spindle area,thechromatin clumped and the centrioles were prominent,the chromosomes were close tothe equatorial plate.The spindle tubules were seen radiating from the centriole andattached to chromosome at the kinetochore,and among them there were a fewmitochondria.In the non-spindle area,the cell organelles could be observed but fewGolgi complex present.It was shown that many changes of the cells could be foundduring mitosis,and that the spindle-fibers,filopodia,rod-like elements and the cellmembranes might play an important role during mitosis.

The effect of cytochalasin D on ECa109 esophageal sqamous cancer cells was studied. When (ECa109) cells were treated with 0.2～1?g/ml cytochalasin D, small epithelial cells with a few microvilli could be seen. After incubation with 0.5?g/ml of cytochalasin D for one or three days, irregular buds containing endoplasmic reticulum, mitochondria and Golgi complexes grew out from the surface of the epithelial cells. The target-shaped mitochondria, the "confronting cisterna" of the endoplasmic reticulum, ring-shaped endoplasmic reticulum and disaggregated Golgi complexes were appeared in the cytoplasm. After treatment with 0.2?g/ml of cytochalasin D for one day multinucleated epithelial cells appeared. There were 2～6 nuclei, with different in size and shape. Some nuclei were fully enclosed with neuclear envelops and the others only partically. Mitotic cells could be seen too. The effect of cytochalasin D on the structure and function of the epithelial cells and the different roles played by the actin and tubulin in keeping the normal function of the cytoskeleton system of the ECa109 cells were discussed.

A comparision of the ECa 109 esophageal epithelial carcinoma cells and normal esophageal epithelial cells was made. Electron microscopically these two cells revealed remarkable differences. The normal esophageal epithelial cells had narrow intercellular space, fewer microvilli, more desmosomes and tonofilaments, with small and round nucleus. No polyribosome could be seen. The ECa 109 cells had wider intercellular space, more microvilli, fewer desmosomes and tonofilaments, larger and irregular nucleus, large and distinct nucleolus. Besides, the appearance of polyribosome was characteristic of cancer cells. Thus, the following characteristics were used as makers of cell differentiation: 1. less microvilli; 2. abundant tonofilaments; 3. more desmosomes; 4. monoribosomes; 5. round nucleus; 6. small nucleus/cytoplasma ratio.Upon addition of 2％ DMSO ECa 109 under went striking morphological changes resembling the terminal differentiation stage of normal esophageal epithelial cells.Inaddition, after DMSO treatment, circular Golgi complexes surrounded by small vesicles could be seen. Small vesicles and primary and secondary lysosomesappeared too. Electron-dense particles with halo were easily seen in the nuclei.

Alkaline Phosphatase ; blood ; Biomarkers ; blood ; Child, Preschool ; Female ; Humans ; Infant ; Isoenzymes ; blood ; Phosphorus Metabolism Disorders ; blood ; diagnosis ; Risk Factors

Objective To analyze the differential expression of protein in taxol-resistance from elderly females with ovarian cancer, by compared proteomics analysis and identified the candidated marker in the clinical tissues. Methods Soluble fraction proteins in two SKOV3 taxol-resistant cells, SK-TR30 and SK-TR2500 and one A2780 taxol-resistant cell, A2780-TR paired taxol-sensitive ovarian cancer cells were separated and prepared by two dimensional gel electrophoresis (2-DE). The differentially expressed proteins were selected and identified by matrix-assisted laser desorption/lonization time of flight mass spectrometry(MALDI-TOF-MS) and database search. 2-DE profiles with high resolution and reproducibility were obtained. Then highly expressed protein-cofilin1 which exist in two forms of de-phosphorylate was detected and compared between human ovarian cancer specimens including 22 elderly female with chemosensitive and 21 elderly female with chemoresistant by immunohistochemistry. Results Among the identified proteins, there were sixteen over/under-expressed proteins in taxol-resistant cell lines as compared with taxol-sensitive cell lines. In particular, overexpression of cofilin1 and destrin were displayed in all three taxol-resistant cell lines. The expression of phosphorylation of cofilin (P＜0.05), but not cofilin1, displayed much higher in chemoresistant than in chemosensitive group. Conclusions Cofilin1 may play a role of taxol resistance by phosphorylation of ovarian cancer cell. However, the mechanisms need to be elucidated further.

Objective:To investigate the effect and the related mechanism of c-Myc on the proliferation,invasion and migration ability of malignant melanoma B16-F10 cells. Methods:We detected the expression of PIK3R3 in malignant melanoma and normal tis-sues. Efficiency of gene silencing of c-Myc and PIK3R3 was determined by qPCR and Western blot. We detected the proliferate ability of B16-F10 cells after silencing c-Myc and PIK3R3 using EdU assay. We detected the migration and invasion ability of B16-F10 cells after silencing c-Myc and PIK3R3 using wound healing assays and Transwell matrigel invasion assays. The expression of miR-199a after silencing c-Myc and PIK3R3 using qPCR. The expression of c-Myc and PIK3R3 was detected by qPCR after transfecting miR-199a mimics or miR-199a inhibitor. Dual-luciferase reporter assay system was used to detect miR-199a regulating c-Myc and PIK3R3. Results:Compared normal skin tissue,expression of PIK3R3 was significantly increased in malignant melanoma tissue;after silencing c-Myc and PIK3R3 gene,the proliferation,invasion and metastasis of melanoma cell line B16-F10 were significantly reduced;expression of miR-199a upregulated after silencing c-Myc and PIK3R3 genes, expression of c-Myc and PIK3R3 decreased after transfecting miR-199a mimics, expression of c-Myc and PIK3R3 upregulated after transfecting miR-199a inhibitor, dual luciferase reporter system test results revealed miR-199a can directly regulate c-Myc and PIK3R3 transcription activity. Conclusion: miR199a regulated the expression of c-Myc,then promote proliferation,migration and invasion in malignant melanoma cells.

Objective To evaluate the blood-saving effect of tranexamic acid in elderly patients undergoing total hip replacement.Methods One hundred and sixty ASA Ⅱ or Ⅲ patientss of both sexes,aged 65-70 yr,with a body mass index of 16-22kg/m2,undergoing total hip replacement,were randomly divided into 2 groups(n =80,each):control group(group C)and tranexamic acid group(group T).Anesthesia was induced with midazolam,fentanyl,etomidate and atracurium.The patients were tracheal intubated and mechanically ventilated.PEr CO2 was maintained at 35-45 mm Hg.Aneslhesia was maintained with propofol,remifentanil and atracurium.Before the skin incision,tranexamic acid 15 mg/kg was infused over 15 m in in group T,while the equal volume of normal saline was given instead in group C.Hemoglobin(Hb),platelet count(PLT),prothrombin time(PT),and activated partial thromboplastin time(APTT)were monitored during operation to guide blood transfusion.Intraoperative and postoperative blood loss and allogeneic blood transfusion were recorded.Postoperative complications were also recorded.Results There was no significant difference in the amount of intraoperative blood loss between the two groups(P ＞ 0.05).The amount of postoperative blood loss was significantly smaller and less allogeneic red blood cell was transfused in group T than in group C(P ＜ 0.05).No complications occurred after operation in either group.Conchusion Tranexamic acid has blood-saving effect in elderly patients undergoing total hip replacement,but the clinical value is limited.

Objective:To investigated the activity inhibition and inhibitory type of polyphenol oxidase (PPO) induced by galangin and the interaction mechanism of galangin with polyphenol oxidase was preliminarily indicated,and prove the related mechanism of galangin on proliferation of on human melanoma A375 cells.Methods: The activity inhibition and inhibitory type of PPO induced by galangin were investigated by spectrophotometric method,and interation mechanism of galangin with PPO was preliminarily indicated by fluorescence quenching and molecular docking,and chelating copper ions with the inhibitory mechanism of galangin on polyphenol oxidase was measured.Results: Galangin was a competitive inhibitor,the IC50 and Ki on PPO were obtained to be (47.86±3.33) and (24.83±1.45)μmol/L,respectively.Fluorescence spectrum indicated the fluorescence of PPO was quenched effectively by galangin and the binding constant Ka was obtained to be (4.67±0.43)×104 L/mol.Chelating copper ions and molecular simulation further showed that galangin was combined with active center of copper ions,and formed hydrogen bonds with catalytic site His259.Luteolin could induce the apoptosis of A375 cells significantly,and the tyrosinase activity and melanin synthesis were decreased.Conclusion: Galangin as a competitive polyphenol oxidase inhibitor and reduced the activity of polyphenol oxidase.which provides the theoretical basis for the clinical anti skin cancer.

Acupuncture Therapy ; Female ; Humans ; Middle Aged ; Preoperative Care ; Tachycardia ; therapy

Obesity-related diabetes and the metabolic syndrome are significantly related to increased adipose tissue mass.Understanding of the effect of adipose tissue on the metabolic regulation will play a key role in therapeutic strategies aimed to reduce obesity-induced insulin resistance and alleviate symptoms of the metabolic syndrome.Visfatin is a novel adipokine that appears to be preferentially produced by visceral adipose tissue and has insulin-mimetic actions.We discusse the relationship between visfatin and metabolic syndrome as well as future therapeutic strategies to these diseases.