Objective Using siRNA to downregulate Her2 expression in Her2 positive breast cancer cell line SK-BR-3,to observe the influence of siRNA of cell proliferation and apoptosis. Methods Two siRNAs against the Her2 gene were designed and transfected into SK-BR-3 cells respectively, Her2 gene expression was measured using RT-PCR, Her2 protein expression was analysed using flow cytometer. Cell proliferation was measured with MTT assay, the apoptotic cells were stained with annexin-V-FITC and analysed by flow cytometry. Results 42.88%,49.27% silencing of Her2 mRNA expression were observed after transfecting with two siRNA respectively. The corresponding decrease of Her2 protein expression was confirmed by flow cytometric analysis. As a consequence, cell growth inhibition was observed,28.51% and 42.81% of cells apoptosis were observed after transfecting with two siRNA respectively.Conclusion These results demonstrated that siRNA against Her2 could effectively downregulate Her2 expression in SK-BR-3 cell line, inhibit cell growth and induce cell apoptosis.

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Objective To investigate the effect of steroid hormones on lymphatic stomata of ovarian bursa and fluid adsorption. Methods The lymphatic adsorption was traced by trypan blue,and the ultrastructure of the lymphatic stomata and estrogen receptor level of mesothelial cells on ovarian bursa was observed by scanning electron microscopy,transmission electron microscopy and immuno-colloid golden technology. Results The absorption and opening area of lymphatic stomata were enhanced in estrogen group mice,and decreased in androgen group as compared to control group mice.Estrogen receptor was first identified in the nuclei of mesothelial cells on the inner layer of mice ovarian bursa and the level was down-regulated by estrogen and slightly up-regulated by androgen.Conclusion The absorption and the opening area of lymphatic stomata can be regulated by steroid hormones in mice,and the machanism may be related to the expression levels of estrogen receptor in the nuclei of mesothelial cells.

Objective To study the ultrastructure of ovary lymphatic stomata in human and compared with the animals′. Methods The morphology of human ovary surface was examined using scanning electron microscopy and the lymphatic stomatas were counted by the ELESCOPE computer image processing system. Results The human ovary epithelium was composed of cuboidal and flat cells. There were lymphatic stomatas not only among the cuboidal cells, but also among the cuboidal and flat cells. The stomata were usually circular in shape and about 1 80?0 82??m in diameter, 6 27?2 65??m in circle. Lymphatic stomata in external surface of ovary mucinous tumor was also found. Conclusion There are ovary lymphatic stomatas in human. These stomatas connect lymphatic vessels in ovary with the peritoneal cavity. They also present morphological basis for early\|metastasis of ovary tumor. [

In fifteen human fetuses spcimens the peritoneal stomata were studied with SEM and TEM, and measured by image processing system. In order to prove that the peritoneal stomata are the passageway of absorbed matter from the peritoneal cavity, animal experiments were made. There are two types of the mesothelial cells on diaphragmatic peritoneum, i. e. the cuboidal cells and the flattened cells. The peritoneal stomata, which arranged in clusters or strips, were only found between the cuboidal cells. The shape and size of the stomata were often irregular. The average area of the stoma on the muscular portion is 10.43?1.61?m~2, while on the tendinous portion is 7.93?1.67?m~2. The connective tissue underlies below the stomata, under which no basement membrane was found. Many lymphatic capillaries were observed in the connective tissue, which may promote absorption of matter from the peritoneal cavity. In animal experiments, some particles of trypan blue were absorbed through the stomata of rabbit diaphragmatic peritoneum. The authors consider that the stomata, are first observed in human, are important pathway for draining matter from the peritoneal cavity.

By transmission electron microscopy and freeze etching technique 15 human fetuses were utilized to study the ultrastructure of the mesothelial cells on the parietal peritoneum. The mesothelial cells of the diaphragmatic peritoneum contained numerous vesicles which were frequently communicated with the free surface, the basement membrane, intercellular space and the peritoneal stomata. Some of the vesicles seemed to fuse each other and form vacuoles. Vacuoles also occurred close to, or communicated with the basement membrane and cell free surface. Sometimes they appeared as secretory particles. The microvilli contained vesicles opened to the free surface. The mesothelial cells on the pelvic wall displayed abundant endoplasmic reticulum and Golgi apparatus, but scanty vesicles. So, the mesothelial cells on parietal peritoneum of human fetuses might be classified into two types, i. e. the vesiclecontaining cells on the diaphragmatic peritoneum and the ER-containing cells on the peritoneum of the pelvic wall. The vesicle-containing cells seemed to uptake material from the peritoneal cavity. Abundant endoplasmic reticulum and Golgi apparatus reflected a high synthetic activity, hence the ER-containing cells might be possibly related to the production of peritoneal fluid.

Objective To investigate the application of delayed sternal closure in the process of open heart surgery.Methods Delayed sternal closure was performed from Mar.2000 to Jun.2005 in 11 patients(4 males and 7 females,aged 16-67 years with an average of 42 years) after cardiac surgery.The indication included: cardiac dilatation(4 cases),intractable arrhythmia(4 cases),continuous bleeding(2 cases) and severe pulmonary edema(1 case).During the open chest period,the wound was covered with 3-layers of latex temporarily,and the delayed sternal closure was performed when bleeding was controlled,heart size reduced,and hemodynamic condition became stable.Results Except two patients died of acute kidney failure during open chest period,delayed sternal closure was successfully carried out in 9 patients 8-43 hours(23.7?11.0) after cardiac surgery.Except one patient died of cardiac arrest 21 days after operation,the 8 others survived and were discharged from ICU 1-8 days(3.9?2.4) after sternal closure,and the wounds healed well.Follow-up for the 8 survivors revealed an improvement of NYHA class(class Ⅰfor 4 cases,class Ⅱfor 2 cases,and class Ⅲ for 2 cases).Conclusion The delayed sternal closure is an effective method in the treatment of postoperative cardiac compression,severe bleeding and arrhythmia,and it does not increase the incidence of complications such as sternal infection.

AIM: To examine the 2′,5′-oligoadenylate synthetase activity and the peritoneal pathological changes when using commercially dialysates in order to reveal effects of the peritoneal dialysate biocompatibility on the immune function and morphological changes. METHODS: (1) CAPD animal models were made using dialysates. (2) Dynamic measurements of 2′,5′-oligoadenylate synthetase activity were made by paper chromatography. (3)The pathological changes of the peritoneal mesothelial cells were examined by light microscope. RESULTS: 2′,5′-oligoadenylate synthetase activities in both experimental (dialysis solution groups) and control group were increased and the mesothelial cell began to exfoliate and become thicker with the infiltration of inflammatory cells at different degrees in the experimental groups, while in the Baxter group, no active inflammation was observed. When the peritoneal dialysis stopped, activity of 2′,5′-oligoadenylate synthetase began to be normal in control group and continued to decline in experimental groups.CONCLUSION: Prolonged immune suppression existed with long-term peritoneal dialysis, causing pathological changes of the peritoneum at certain degrees. Different dialysis solutions resulted in different degrees of immuno-supression and peritoneal damages, which was relevant to the biological compatibility of the peritoneal dialysate.

AIM: To study the regulatory effect of nitric oxide on the lymphatic stomata and probe into the mechanism of ultrafiltration failure during long-term peritoneal dialysis (PD). METHODS: ①Sodium nitroprusside(SNP) and N G-monomethyl-L-arginine(L-NMMA) (inhibitor of nitric oxide synthase(NOS)) were injected into the peritoneal cavity of the mouse model of PD. ②NO concentration was measured in serum. ③The lymphatic stomata was studied with SEM and computer image processing.RESULTS: During PD, a lot of macrophages wandered out of the lymphatic stomata to form milky spots on the peritoneal mesothelium, and the diameter and density of the stomata were increased with NO concentration raised. After PD cessation, the stomata was normal gradually and numbers of milky spots reduced with NO concentration fall. The diameter and density of the stomata were increased with a rise in NO concentration as SNP was used, oppositely those were decreased with the increase in NO concentration as L-NMMA was injected intraperitoneally. CONCLUSIONS: The lymphatic stomata might be regulated through increasing the endogenous NO concentration. During PD, NO is increased gradually and the ultrafiltration failure would occur when re-absorption of the stomata was increased from the peritoneal cavity.

AIM: To investigate the expression of vascular endothelial growth factor receptor-3 (VEGFR-3) and CD31 in relation to metastasis in ovarian epithelial tumors. METHODS: VEGFR-3 and CD31 expression were examined by immunohistochemical methods, VEGFR-3 positive microlymphatic count (MLC) and microvessel density (MVD) were assessed by the image analysis. RESULTS: Both MLC and MVD in ovarian epithelial carcinomas were higher than those in benign tumors(MLC, P 0 05). CONCLUSIONS: VEGFR-3 and CD31 expression correlate significantly with metastasis, MLC and MVD might predict peritumoral lymphangiogenesis and angiogenesis, which may be as a biologic marker for metastasis in ovarian epithelial tumors. [