Objective To construct a cDNA library of human epithelial ovarian carcinoma tissue for screening ovarian carcinoma specific-antigen.Methods The total RNA was separated from human epithelial ovarian carcinoma tissue.The mRNA from total RNA was isolated to synthesize the first and second strand cDNA.The ds-cDNA termini were blunted with pfu DNA polymerase.The blunted cDNAs were added EcoR Ⅰ adaptor and then digested by XhoⅠ.Small cDNA molecules(less than 400 bp) were removed through size fraction.After the cDNAs were ligated into ZAP expression vector,the ligated products were packaged in vitro and the bacteriophage particles infected the host strains XL1-Blue MRF′.Results The efficiency of the primary library was 5.5?10~(6)pfu/ml and the amplified library was 3.0?10~(11)pfu/ml with 96% clones positive.The average length of the inserted fragment was over 1 kb.Conclusion The quality of the constructed human epithelial ovarian carcinoma tissue cDNA library is excellent and helpful to screen ovarian carcinoma specific-antigen.

Objective:Human capillary lymphatic endothelial cells (LECs) were isolated and cultured to assist further investigation of the function of lymphatic vessels generation during cancer metastasis. Methods:Human skin was digested by type I collagenase. Cells were isolated using magnetic beads which were marked by monoclonal antibodies against the extracellular domain of VEGFR-3, and purified by cloning col-umn. The morphology and structure of cells were observed by microscopy. Immunophenotype was identified by immunofluorescence. Cell growth curve was recorded to measure the effect of VEGF-C protein. Results:LECs exhibited the typical cobblestone morphology as monolayer growth pattem under microscopy. Enlarged nucleus and rich cytoplasm with bubbles were found under electromicroscopy. LECs specific markers inclu-ding VEGFR-3, LYVE-1, Podoplaninand D2-40 all were positive. VIII factor as specific marker of blood ves-sel endothelium cells (BVECs) was negative. VEGF-C induced a marked increase of cell proliferation. Con-clusions:Human dermal LECs could be harvested successfully using collagenase digestion, immunomagnet-ic beads sorting and clonic column purification.

Objective To observe the clinical behaviors of cervical intraepithelial neoplasia (CIN) for early diagnosis to improve the therapeutic efficacy Methods Eighty seven cases with different grades of CIN confirmed with colposcopy and pathological examination were reviewed for the clinical features and the results from colposcopy, pathology, testing for HPV (human papillomavirus) All patient were treated with various methods and followed up Results Most patients with CIN were in childbearing period and sought for medical advices because of increased leucorrhagia and/or bloody leucorrhea, postcoital bleeding for a comparatively long course Chronic cervicitis was often found in these patients Contact bleeding and HPV infection were common in patients with CINⅡand CIN Ⅲ Patients with CIN Ⅰ were regular followed up or treated with electrocautery, those with CIN Ⅱ were treated with conization or hysterectomy, and for those with CIN Ⅲ, hysterectomy was performed The outcomes of all patients were satisfactory after treatment Conclusion The most common sign for the patients with CIN is postcoital bleeding For correct diagnosis of CIN, colposcopy and together pathological examimotion is recommended High risk HPV detection is a good prognostic factor After treatment, the patients with different grades of CIN should be followed up regularly

Objective To evaluate the value of multi-slice CT(MSCT)classification in the assessment of the hilar cholangiocarcinoma resectahility.Methods Thirty patients with surgically and histopathologically proved hilar cholangiocarcinomas who underwent preoperative MSCT and were diagnosed correctly were included in present study.Transverse images and reconstructed MPR images were reviewed for Bismuth-Corlette classification and morphological classification of hilar cholangiocarcinoma.Thcn MSCT classification was compared with findings of surgery and histopathology.Curative resectabilty of different types according to Bismuth-Corlette classification and morphological classification were analyzed with chi-square test.Results In 30 cases,the numbers of Type Ⅰ,Ⅱ,Ⅲa,Ⅲb and Ⅳ according to BismuthCorlette classification were 1,3,4,5 and 17.Seventeen patients underwent curative resections,among which 1,2,1,4 and 9 belonged to Type Ⅰ,Ⅱ,Ⅲa,Ⅲb and Ⅳ respectively.However,there was no significant difference in curative resectability among different types of Bismuth-Corlette classification(X2=0.9875.P＞0.05).In present study,the accuracy of MSCT in Bismuth-Corlette classification reached 86.7%(26/30).The numbers of periducatal infiltrating,mass forming and intraductal growing type were 13,13 and 4,while 6,8 and 3 cases of each type underwent curative resections.There was no significant difference in curative resectability among different types of morphological classification(X2=1.2583,P＞0.05).The accuracy of MSCT in morphological classification was 100%(30/30)in this study group.Conclusion MSCT can make accurate diagnosis of Bismuth-Corlette classification and morphological classification.which is helpful in preoperative respectability assessment of hilar cholangiocarcinoma.

Objective To investigate the imaging and biodistribution of 99Tcm labeled peptide ( phage-SHSWHWLPNLRHYAS) high-binding VEGF receptor 3 in nude mice with ovarian cancer. Methods We used NHS-MAG3 as the bifunctional chelating agent and synthesized the peptide above. It was completed with pre-stannous direct labeling by 99Tcm and then the labeling efficiency was determined by paper chromatography. The tumor bearing mice were injected via tail veins with radiolabeled peptide. The tumors were imaged with single photon emission computed tomography ( SPECT) . We measured and calculated the biodistribution of the radiolabeled peptide. Results The labeled rate was 95. 27% and the radiochemical purity was 96% . Under a SPECT apparatus,we observed tumor location image at 1 h post injection and the tumor images was the clearest at 3 h post injection,with the injected dose per gram of tissue ( % ID/g) of ( 30. 20 ? 6. 89) at the tumor sites. The tumor/muscle ratio was 13. 13. Conclusion Our peptide can accumulate in the tumor masses. It is hopeful to be used as an ovarian cancer targeting vector for diagnosis and therapy of the cancer.

Objective To investigate the correlation between the expressions of vascular endothelial growth factor C (VEGF C) and vascular endothelial growth factor D (VEGF D) and the lymph node metastasis of groin and retroperitoneum in human epithelial ovarian carcinoma. Methods The expressions of VEGF C and VEGF D in benign, borderline, and malignant epithelial ovarian tumors were detected by immunohistochemistry and light microscopy. Results The expressions of VEGF C and VEGF D were found in benign, borderline, and malignant epithelial ovarian tumors. The expressions of VEGF C and VEGF D in ovarian carcinomas were significantly higher than those in benign and borderline ovarian tumors. The expression levels of VEGF C and VEGF D were correlated with peritoneal metastasis, lymph node metastasis of groin and retroperitoneum, and clinical stage, but not with distant metastasis, histology type, histological grade, and age. Conclusion Up regulation of VEGF C and VEGF D in epithelial ovarian carcinoma is advantageous to lymph node metastasis. This process may probably be correlated with lymphangiogenesis in tumors.

BACKGROUND:Bone marrow mesenchymal stem cells(BM-MSCs)have been shown to possess the potential to differentiate into oocytes.However,immune rejection and a limited number of donors of BM-MSCs constrain the applications of BM-MSCs.Several studies have demonstrated that human umbilical cord matrix stem cells(UC-MSCs)also have an intrinsic ability to differentiate into oocyte-like cells in vitro.OBJECTIVE:To establish the method for UC-MSCs culture and to investigate the in vitro differentiation potential of UC-MSCs towards germ cells.METHODS:Umbilical cord from full-term normal deliveries was obtained in sterile condition.Collagenase I-digested cells were cultured in DMEM.The immunophenotype of cells was determined by flow cytometry.Lipoblasts,osteoblasts and chondroblasts were induced in different condition cultures.The expression of germ cells specific marker in UC-MSCs was determined by reverse transcdption-polymerase chain reaction.Follicular fluid was employed to induce UC-MSCs differentiation into germ cells.RESULTS AND CONCLUSION:Spindle-like umbilical cord cells were shown and cells in culture were extended to more than 10passages.BM-MSCs-like immunophenotypes were shown:CD29,CD44,CD73(SH3),CD90 and CD105(SH2)were positive;SSEA-4 was weakly positive;CD31,CD34,CD45 and HLA-DR were negative.After UC-MSCs were induced in different condition cultures,lipid droplet-,bone tubercle-,and cartilage tubercle-like structures emerged and the mRNA expressions of specific gene of fat,bone and cartilage were observed.Germ cells markers,OCT4,Stella,Ifitm3,were expressed in UC-MSCs.After induced by 5%,10% or 20% follicular fluid,cells aggregated and oocyte-like structures were observed.Human UC-MSCs could be cultured and amplificated in vitro.UC-MSCs showed immunophenotypes similar to BM-MSCs.UC-MSCs had the potential to differentiate into lipoblasts,osteoblasts,and chondroblasts.Oocyte-like structure was induced in vitro from UC-MSCs with germ cells specific marker.These findings suggest that UC-NSCs have the potential to differentiate into germ cells.

Objective To investigate clinical features, diagnosis, and treatment of refractory Mycoplasma pneumoniae pneumonia (MPP) combined with thrombosis. Methods One case with refractory MPP associated with thrombosis was retrospectively analyzed with literature review. Results The patient presented with respiratory symptom at the onset, while melosalgia and decreased respiratory sound in left lung were occurred during anti-infection therapy. Thereafter, thrombosis of lower extremity veins and pulmonary embolism were confirmed by a series of examinations. Serum anti-phospholipid antibody was positive. Finally, the patient was treated with a combination of anticoagulation and immunosuppressive therapies. Conclusions The mechanism of refractory MPP combined with thrombosis may be associated with excessive inflammatory response and endothelial cells injury. The thrombosis complication should be suspected in patient of Mycoplasma pneumonia infection with positive anti-phospholipid antibody and low concentration of protein C and immunosuppressive therapy should be implemented promptly.

Objective To investigate the ability of electrocardiogram-gated multislice CT(MSCT)in the diagnosis of myocardial bfidging.Methods Fifty-one patients(82 coronary arteries)with suspected coronary artery disease underwent multi-detector row CT,conventional coronary angiography and intravascular ultrasonography as well.The sensitivity,specificity and accuracy of MSCT for the detection of myocardial bridging were determined.The interobserver agreement was calculated by using Cohen's Kappa test.Results A total of 26 tunneled arteries exclusively located near the middle segment of left anterior descending coronary artery were found by coronary angiography and intravascular uhrasonography.Compared to the invasive methods,MSCT correctly detected 23 of 26 myocardial bridges with a sensitivity of 88%(23/26),specificity of 96%(52/54)and accuracy of 94%(75/80).The Kappa value for overall interobserver variation Was 0.62.Two myocardial bridges diagnosed by MSCT were missed with the invasive method.With the results of invasive and non-invasive methods combined as the standard of reference,the overall sensitivity.specificity,and accuracy of MSCT in detecting myocardial bridging were 89%(25/28),91%(21/23),and 90%(46/51),respectively.Conclusion As a non-invasive imaging modality,MSCT is feasible and reliable in the detection of myocardial bridging.