Hes1/Hes5 gene inhibits differentiation via down-regulating Hash1 and promotes proliferation in cervical carcinoma cells.

Introduction: Hairy and enhancer of split 1 (Hes1) and Hes5 are target genes for the mammalian Notch pathway, which are highly expressed in epithelia in the process of embryogenesis or in neural stem cells, inhibit cell differentiation via the Notch-Hes-Hash signaling, and promote the survival of stem cells. Either Hes1 or Hes5 overactivation is likely to affect cell differentiation, thereby resulting in carcinogenesis.

Methods: We transfected the diced small interference RNA into SiHa cells and detected cell differentiation and proliferation by immunocytochemistry, Western blot, and methyl thiazolyl tetrazolium assay.

Expression of octamer-4 in serous and mucinous ovarian carcinoma.

Background: Octamer-4 (Oct4) is a well known regulator of self-renewal in embryonic stem cells; it has been detected in several human cancers and may play a critical role in carcinogenesis.

Aims: To assess the expression of Oct4 in epithelial ovarian tumours.

Methods: Expression of Oct4 was evaluated by immunohistochemistry in 460 cases of various epithelial ovarian lesions as well as 35 cases of normal fallopian tube epithelium.

Methotrexate induces apoptosis of human choriocarcinoma cell line JAR via a mitochondrial pathway.

Objective: To investigate methotrexate (MTX)-induced apoptosis and the involved pathways in human choriocarcinoma cells.

Study Design: MTX-induced apoptosis of human choriocarcinoma cell line JAR was examined using a PI/Annexin V stain with flow cytometer (FCM). Mitochondrial apoptosis was detected by fluorescence microscopy, and analyzed by FCM using a MitoCapture mitochondrial apoptosis detection kit.

[Study of apoptosis and Fas expression of peritoneal fluid and peripheral blood T lymphocytes in patients with epithelial ovarian cancer and their relationship with CA125].

Objective: To investigate the apoptosis and Fas (CD95) expression of T lymphocytes from the peripheral blood and peritoneal fluid of the patients with ovarian cancer and their relationship with CA125.

Methods: Apoptosis and Fas expression of peritoneal fluid and peripheral blood T lymphocytes were assessed by flow cytometry. Peripheral blood samples were obtained from the following objects respectively: patients with stage III - IV ovarian cancer (n = 18) before and after treatment, patients with stage I - II ovarian cancer (n = 15), patients with benign ovarian tumor (n = 18), patients with Krukenberg tumor (n = 6) and normal control (n = 20).

Fas-mediated pathway and apoptosis in normal cervix, cervical intraepithelial neoplasia and cervical squamous cancer.

The aim of this study was to examine the expression patterns of apoptosis-related antigens, such as Fas, FasL, and cFLIP, in cervical squamous cells, and investigate the role of Fas-mediated apoptosis in the pathogenesis of cervical neoplasia. Using specific antibodies for Fas, FasL, and cFLIP, we examined protein expression in 19 specimens of normal cervix, 15 mild dysplasia (CIN I), 22 moderate dysplasia (CIN II), 23 severe dysplasia (CIN III), and 34 invasive squamous cell carcinoma (SCC) by immunohistochemistry. We detected the apoptotic indices by TUNEL in the same specimens.

[Deletion of OPCML gene and promoter methylation in ovarian epithelial carcinoma].

Objective: To investigate the expression of OPCML gene in ovarian epithelial carcinoma and determine the relationship between mRNA expression and methylation of their promoters.

Method: Twenty normal ovarian tissues and 89 ovarian epithelial tumor specimens (72 malignant, 17 benign), as well as 3 ovarian carcinoma cell lines (SKOV-3, CAOV3, and 3AO), were collected for detection of OPCML gene expression by reverse transcription-polymerase chain reaction and for detection of promoter methylation by restriction enzyme cut analysis from 7. 1999 to 7.

[The accuracy of diagnosing cervical intraepithelial neoplasia with colposcopically directed biopsy].

Objective: To evaluate the accuracy of colposcopically directed biopsy in the diagnosis of cervical intraepithelial neoplasia (CIN).

Methods: The clinical data of 153 patients with CIN diagnosed by colposcopically directed biopsy and treated with cervical loop electrosurgical procedure (LEEP) shortly after were retrospectively studied. The consistency of pathological examination between colposcopically directed biopsy and LEEP was evaluated.

[Evaluation of neoadjuvant chemotherapy for locally advanced cervical cancinoma].

Objective: To investigate the effect of neoadjuvant chemotherapy on locally advanced cervical cancer, the factors affecting outcome of chemotherapy and the long-term survival rate in the patients.

Methods: A total of 64 patients with stageIb2-IIb of locally advanced cervical cancers treated with neoadjuvant chemotherapy between June 1999 and October 2004 in Women's Hospital, School of Medicine, Zhejiang University were retrospectively analysed. The effect of chemotherapy, as well as survival rate was evaluated.

Altered expression of cellular membrane molecules of HLA-DR, HLA-G and CD99 in cervical intraepithelial neoplasias and invasive squamous cell carcinoma.

Objective: The aim of this study was to investigate the role of HLA-DR, HLA-G and CD99 during cervical carcinogenesis and to examine the prognostic significance of these protein expressions in invasive squamous cell carcinoma (SCC).

Methods: Using specific antibodies for HLA-DR, HLA-G and CD99, we examined protein expressions in 19 normal cervix, 15 mild dysplasia (CIN I), 22 moderate dysplasia (CIN II), 23 severe dysplasia (CIN III), and 34 invasive squamous cell carcinoma by immunohistochemistry. And we detected the expression of Ki67 in the same specimens.

Proportion of CD4+CD25+ regulatory T cell is increased in the patients with ovarian carcinoma.

Objective: To study the frequency of the CD4+CD25+ regulatory T cells (Tregs) in the patients with ovarian carcinoma and its possible mechanism.

Methods: The percentages of CD4+CD25+ Tregs in the peripheral blood lymphocytes (PBLs), tumor infiltrating lymphocytes (TILs) and tumor associated lymphocytes (TALs) from 13 patients with ovarian carcinoma and in the PBLs from 14 healthy women were determined by flow cytometry. The expression of CD69 on CD4+PBLs from the patients was detected.

Ovarian carcinoma cells effectively inhibit differentiation and maturation of dendritic cells derived from hematopoietic progenitor cells in vitro.

Loco-regional dissemination of ovarian carcinoma is associated with immunosuppression of the peritoneal cavity. One marked characteristic of the peritoneal immunity in this disease is the defective function of dendritic cells (DCs). In this study, the affect of ovarian carcinoma cells on DCs derived from hematopoetic progenitor cells was observed.

[Tumor lymphangiogenesis in cervical squamous cell carcinoma and its clinical significance].

Objective: To investigate the clinical significance of lymphangiogenesis in cervical cancer.

Methods: Monoclonal podoplanin was used to immunostain the lymphatic microvessels in the paraffin sections of cervical squamous cancer tissues at the I(b) and II(a) stages from 35 cases kept in the archives. Computer-assisted morphometric analysis was used to quantitatively analyze the lymphatic vessels in intratumor and peritumor areas.

[Tumor-infiltrating dendritic cells in epithelial ovarian carcinoma and correlation with the expression of vascular endothelial growth factor].

Objective: To investigate the density and activation status of tumor infiltrating dendritic cells (TIDC) in epithelial ovarian carcinoma (EOC) and correlation with the expression of vascular endothelial growth factor (VEGF).

Methods: Streptavidin-peroxidase (SP) and Picture two-step immunohistochemistry methods were used to detect S-100(+), CD(83)(+) TIDC and the expression of VEGF in 57 primary EOCs, 32 benign ovarian tumors (benign control) and 16 normal ovarian tissues (normal control).

Results: (1) Two types of heterogeneous distribution pattern of TIDC in EOC were observed under the microscope.

[Analysis of prognostic factors in patients with cervical squamous cell carcinoma of stage Ib and IIa].

Objective: To analyze the prognostic factors in patients with cervical squamous cell carcinoma of stage Ib and IIa treated by surgery, and to investigate their guide roles in available post-operation adjuvant therapy.

Methods: The clinicopathologic records of 306 patients with cervical squamous cell carcinoma of stage Ib and IIa who underwent radical hysterectomy and pelvic lymphadenectomy were retrospectively analyzed, and the prognostic factors were explored by univariate and multivariate methods. Independent prognostic factors were identified by COX proportional hazards regression model.

[Expressions of VEGF/VEGFRs and activation of STATs in ovarian carcinoma].

Objective: To study the expressions of VEGF/VEGFRs and activation of STATs in ovarian epithelial carcinoma, and to elucidate direct effect of VEGF on ovarian carcinoma cells.

Methods: Tissue samples from 42 women with primary ovarian epithelial carcinoma (OVCA), 29 with begnin ovarian tumor (OVBT) and 11 with normal ovarian tissue (NOV) were collected. LSAB immunohistochemical staining was used to determine the expression of VEGF, VEGFR1, VEGFR2 and activated STATS (P-STAT1, P-STAT3, P-STAT5, P-STAT6) proteins.

[Establishment of two-dimensional electrophoresis of uterine leiomyoma for the proteome analysis].

Objective: To establish and optimize the two-dimensional electrophoresis (2-DE) of uterine leiomyoma for the proteome analysis.

Methods: Run immobilized pH gradient (IPG)-isoelectric focusing electrophoresis as the first dimension, then vertical SDS-PAGE electrophoresis as the second dimension. A series of important steps,such as sample solubility, volume of loading, electrophoresis parameters and protocol for staining were optimized.

[Effects of vascular endothelial growth factor on differentiation and function of dendritic cells generated from CD34+ hematopoietic progenitor cells in vitro].

Objective: To investigate the effects of vascular endothelial growth factor (VEGF) on differentiation and function of dendritic cells derived from CD34+ hematopoietic progenitor cells.

Methods: After isolation from umbilical cord blood with a high-gradient magnetic cell sorting system (MACS), the CD34+ cells were cultured with a cocktail cytokines for differentiating into dendritic cells (DC). The cells were stimulated by VEGF (25 ng/ml) either at the beginning or at day 9 of culture.

[Development of HPV16 positive cervical cancer model in the hu-PBL-SCID mouse and its immunological features].

Objective: To develop a HPV16 positive cervical cancer model in the hu-PBL-SCID mouse and investigate its immunological features.

Methods: Thirty-two CB17SCID mice were randomly divided into 4 groups: group A (5 mice) subcutaneously injected with phosphate-buffered saline, group B (5 mice) intraperitoneally injected with human peripheral blood lymphocyte (PBL) for immune reconstruction, group C (11 mice) subcutaneously injected with human cervical carcinoma cell line SiHa, and group D (11 mice) intraperitoneally injected with PBL and subcutaneously injected with SiHa cells after 24 hours of PBL transplantation. The tumor growth, behaviors and status of xenogeneic graft versus host disease (XGVHD) were observed.

[Phosphorylation of signal transducer and activators of transcription 3 induced by vascular endothelial growth factor in ovarian carcinoma cell line in vitro].

Objective: To observe phosphorylation of signal transducer and activators of transcription 3 (STAT3) in Caov-3 induced by vascular endothelial growth factor (VEGF), and to investigate molecular mechanisms of the effect of VEGF on ovarian carcinoma cells.

Methods: The expressions of phosphorylated STAT3 in Caov-3 induced by VEGF were detected by immunocytochemistry and Western blot methods. Furthermore, the relationship between STAT3 phosphorylation and VEGF stimulation in ovarian carcinoma cells was investigated using a peptide which could specifically bind VEGFR2 and thus block the binding of VEGF to its receptors.

Vascular endothelial growth factor (VEGF) and ovarian carcinoma cell supernatant activate signal transducers and activators of transcription (STATs) via VEGF receptor-2 (KDR) in human hemopoietic progenitor cells.

Objective: To investigate the STATs signaling pathway activated by VEGF in human hemopoietic progenitor cells.

Methods: CD34(+) hemopoietic progenitor cells, which isolated from umbilical cord blood, were treated with VEGF or culture supernatant of ovarian carcinoma cell line which could secrete large amount of VEGF, phosphorylation and nuclear translocation of STAT3 and STAT5 were then detected by Western Blot and immunocytochemistry. Expression of VEGFR2/KDR on CD34(+) cells was studied by immunocytochemistry.

[Relationship between interleukin-10 level in ascites of patients with primary ovarian epithelial carcinoma and immune defect in peritoneal cavity].

Background & Objective: As a multifunctional Th2-cytokine, interleukin-10 (IL-10)plays a major role in the immune response. It is well known that IL-10 is an immunosuppressive cytokine, and participates in the development and progression of various tumors. In this study, we investigated the relationship between the IL-10 level in the ascites of the patients with primary ovarian epithelial carcinoma (POEC) and immune defect in the peritoneal cavity.

[Analysis of the risk factors for ovarian metastasis in patients with endometrial carcinoma].

Objective: To study the risk factors for ovarian metastasis in patients with endometrial carcinoma.

Method: The pathological and clinical features and outcomes of endometrial carcinoma patients who were diagnosed and treated in our hospital from Jan 1996 to Dec 2002 were retrospectively reviewed and analyzed.

Results: Of the 321 cases reviewed, 15 (4.

[Establishment of methotrexate-resistant human choriocarcinoma cell line and its biological characteristics].

Objective: To establish a methotrexate (MTX)-resistant choriocarcinoma cell line and to determine its biologic characteristics.

Methods: MTX-resistant cell line (JAR/MTX) was derived from human choriocarcinoma cell line JAR by exposed to intermittently and progressively increasing concentration of MTX. Drug sensitivity was detected by MTT; P-gp GST-Pi and PCNA expressions were detected by immunohistochemistry.

[Activation of signal transducers and activators of transcription induced by vascular endothelial growth factor in CD34+ hematopoietic progenitor cells in vitro].

Objective: To investigate the activation pattern of signal transducers and activators of transcription (STAT) induced by vascular endothelial growth factor (VEGF) in CD34+ hematopoietic progenitor cells, and gain an insight into the molecular mechanism and signal transduction pathway of VEGF that has an effect on CD34+ hematopoietic progenitor cells.

Methods: After isolated from umbilical cord blood by using a high-gradient magnetically activated cell sorting system (MACS), CD34+ cells were stimulated by VEGF (50 ng/ml) for different time (0, 15, 30, 45, 60, 90 min) to detect the tyrosine phosphorylation and nuclear translocation of STAT-3 and STAT-5 with Western blot and immunocytochemistry methods. The expression of VEGF receptor-2 (VEGFR2) on the membrane of CD34+ progenitor cells was examined by immunocytochemistry.

Effect of interleukin-7 gene transfection into ovarian carcinoma cell line SKOV3 in vitro and in vivo.

Objective: To investigate the effect of interleukin-7 (IL-7) gene transfection into an established ovarian carcinoma cell line (SKOV3) in vitro and evaluate the tumorigenicity of SKOV3-IL-7 in severe combined immunodeficient (SCID) mice.

Methods: IL-7 gene was transfected into SKOV3 cells by liposome. IL-7 mRNA and protein of SKOV3-IL-7 and their parental control cells were detected by reverse transcriptive-polymerase chain reaction (RT-PCR) and Western blot, respectively.