Search

A series of novel Ln8 clusters show highly heterogeneous catalytic activity for catalyzing the cycloaddition of CO2 with epoxides under mild conditions. Importantly, the Gd8 cluster exhibits a significant MCE with a larger ΔSm of 46.14 J kg−1 K−1.

The combination of fenretinide and selenite on ovarian cancer cells was investigated to assess its effects on proliferation and ability to induce apoptosis. Our results showed that fenretinide and selenite in combination significantly suppress the proliferation of ovarian cancer cells and induced apoptosis (including reactive oxygen species generation, and the loss of mitochondrial membrane potential) compared with either drug used alone. The caspase3/9-dependent pathway was triggered significantly in combination treatment, and moreover, the AMPK pathway also mediated the apoptosis induction in fenretinide and selenite combination. Fenretinide and selenite combination treatment was demonstrated to suppress tumor growth in vivo, this drug combination has been thus found to have an enhanced anti-tumor effect on ovarian cancers cells.

BACKGROUND: A previous study demonstrated that local application of levonorgestrel-loaded polylactic acid microspheres (LNG microspheres) resulted in significant regression of endometriotic cysts in a rabbit model for 6 months without disturbing the metabolic parameters or ovarian function. In order to investigate the feasibility of local application of LNG microspheres as a long-term maintenance treatment for endometriosis, the suppressive effect of a single intra-cystic injection of LNG microspheres was studied for 1 year in a rat model. METHODS AND RESULTS: Twenty four rats with experimental endometriotic cysts were randomized to be treated with a single intra-cystic injection of LNG microspheres (n = 8); 6-month GnRH agonist (GnRHa, n = 8) or control (n = 8). Intra-cystic injection of LNG microspheres and GnRHa treatment caused comparable regression and atrophy in endometriotic cysts in the first 6 months. Compared with the control, the wet weight of the endometriotic cysts was significantly lower in both groups at Month 6 but by Month 12 only remained low in the LNG microspheres group (P < 0.01). The immunostaining of estrogen receptors (ERs) in both the epithelium and stroma and progesterone receptors (PRs) in the stroma was significantly weakened in the LNG microspheres group at Month 6 and was not fully restored at Month 12 (P < 0.01). Metabolic parameters and estrous cycle were not disturbed by local application of LNG microspheres. CONCLUSIONS: In a rat endometriosis model, the suppressive effect of a single intra-cystic injection of LNG microspheres was comparable to that of GnRHa, and was maintained for 1 year. The down-regulation of ERs and PRs might serve as possible mechanism of long-term effectiveness.

Aurora-A, a novel member of the serine/threonine kinase family, has been reported to be correlated with tumorigenesis. Our aim was to investigate whether Aurora-A expression correlates with clinicopathologic factors and prognosis of cervical carcinoma patients.Retrospective study.Seventy-four cervical carcinoma patients, between 1996 and 2002.Reverse transcription-polymerase chain reaction and Western blot assays were performed to detect the expression of Aurora-A gene in cervical carcinoma cells and paired cancerous and corresponding noncancerous tissues from 74 cervical carcinoma patients. The expression of Aurora-A protein in tissues was also determined by immunohistochemistry. The relationships of Aurora-A expression with clinical factors and prognosis of patients were evaluated by statistical analysis.The expression of Aurora-A mRNA and protein was significantly higher in cervical carcinoma cells than in normal cervical epithelial cell (p0.05). The expression of Aurora-A mRNA in cancerous tissues was significantly higher than that in corresponding noncancerous tissues (p0.001). The expression of Aurora-A protein was also increased in tumor tissues by immunochemistry. Aurora-A transcript expression was correlated with FIGO stage (p=0.018), tumor differentiation (p=0.014), parametrial invasion (p=0.024), lymphnode or hematogenous metastasis (p=0.005 or 0.019), but not other clinicopathological factors. Patients with high Aurora-A expression had a poorer disease-free survival and overall survival rates than patients with low Aurora-A expression. Multivariate analysis showed that high Aurora-A mRNA expression was an independent prognostic factor (risk ratio: 2.88; p=0.005).Aurora-A might be used as a prognostic marker for cervical carcinoma patients.

To investigate the inhibiting mechanism of multi-drug resistance (MDR) using expression vectors of short hairpin RNA (shRNA) in a MDR human ovarian cancer cell line (A2780/cp70).Two shRNA expression vectors were constructed and introduced into A2780/cp70 cells. Expression of MRP2 mRNA was assessed by RT-PCR, and Mrp2 expression was determined by Western blot and immunocytochemistry. Apoptosis and sensitization of the cells to cisplatinum were quantified by flow cytometry and methyl-thiazol-tetrazolium (MTT) assays. Cellular cisplatinum accumulation was assayed by laser scanning confocal microscopy (LSCM).In A2780/cp70 cells transfected with MRP2-A and MRP2-B shRNA expression vectors, RT-PCR showed that MRP2 mRNA expression was reduced by 41.8% (P0.05), 30.9% (P0.01) (transient transfection) and 39.6% (P0.05), 29.4% (P0.01) (stable transfection), respectively. Western blot and immunocytochemistry showed that Mrp2 expression was significantly and specifically inhibited. Resistance against cisplatinum was decreased from 173- to 119-fold (P0.05), 64-fold (P0.01) (transient transfection) and to 117-fold (P0.05), 60-fold (P0.01) (stable transfection). Furthermore, shRNA vectors significantly enhanced the cellular cisplatinum accumulation. The combination of shRNA vectors and cisplatinum significantly induced the apoptosis of cells.shRNA expression vectors effectively reduce MRP2 expression and can restore the sensitivity of drug-resistant cancer cells to conventional chemotherapeutic agents.

Survivin is a new member of the inhibitors of apoptosis (IAPs) family. It is upregulated in various malignancies including human cervical carcinomas. Reduction of this molecule has resulted in chemosensitization, but it is uncertain whether it can lead to radiosensitization. We observed the effect of survivin gene RNA interference (RNAi) on the proliferation, apoptosis, and radiosensitivity of the human cervical carcinoma cells HeLa.Human cervical carcinoma cells (HeLa) were transfected with the specific siRNA expression vector (pSilencer2.1-s2) designed to target survivin mRNA. A corresponding site-mutated vector was constructed as a negative control (pSilencer2.1-NC). The expression of survivin mRNA and its protein among the stable transfected cells and the untransfected ones was detected by semi-quantitative RT-PCR and Western blotting respectively. The cell growth was examined by methyl thiazolyl tetrazolium (MTT) assay. The cell cycle distribution and cell apoptosis were measured by flow cytometry. The changes in cell radiosensitivity were observed by clonogenic survival assay.Three stable transfected cell lines: HeLa-s2 (with pSilencer2.1-s2), HeLa-NC (with pSilencer2.1-NC), and HeLa-U6 neo (with empty vector pSilencer2.1-U6 neo) were established. The expression levels of survivin gene mRNA and protein in HeLa-s2 were significantly lower than in HeLa-NC, HeLa-U6 neo, and those untransfected HeLa cells. The expression inhibitory rates were 62.8% and 60.1%. The cell proliferation of HeLa-s2 was inhibited, and the highest inhibitory rate was 57.8+/-2.1%. The changes in cell cycle distribution in HeLa-s2 compared with the other three cell lines were obvious, many cells were blocked in the G(0)/G(1) phase 72.7+/-3.1% (P0.05), reduced sharply in the G(2)/M phase (5.1+/-2.9)% (P0.05), and also the apoptotic rate was 29.2+/-1.4%, obviously increasing (P0.05). At the same dose of radiation, the cloning efficiency of HeLa-s2 declined notably (P0.05); the cell survival curve showed a significant decrease in D(0) and D(q), which were 3.15 and 1.21, respectively (P0.05), and the radiation enhancement ratios were 2.01 (a ratio of D(0)) and 1.77 (a ratio of D(q)).Survivin gene RNAi not only could inhibit the proliferation of human cervical carcinoma cells (HeLa), but also could significantly enhance the radiosensitivity of those cells through the reduction of its mRNA and protein. Therefore, an RNAi-targeted survivin gene strategy would be a potential approach to radiosensitization therapy in human cervical carcinomas.

Objective To explore the expression of urokinase-type plasminogen activator(uPA) in epithelial ovarian carcinoma(EOC) and the relationship of the expression to microvessel density.Methods SP immunohistochemical staining was performed to determine the expression of uPA in ovarian cancer(85 cases),borderline ovarian tumor(16 cases),benign ovarian tumor(39 cases) and normal ovarian tissue(24 cases).Microvessels in ovarian cancer were marked by CD34,and microvessel density(MVD) was determined by direct count.The relationship between uPA and MVD was analyzed.The 85 patients with epithelial ovarian cancer were followed up.Results The highest positive rate of uPA existed in the patients with EOC.There was a correlation between the expression of uPA and MVD and the differentiation of EOC,clinical stage,lymphatic metastasis,omental metastasis,and 5 years survival rate.A significant positive correlation was found between the expression of uPA in EOC and MVD(r=0.56,P=0.02).Conclusion uPA may promote the angiogenesis of EOC,and participate in the occurrence,development,invasion and metastasis of ovarian cancer.The detection of uPA and MVD may be used as an indicator of biological behaviors and prognosis of ovarian cancer.

Summary PTEN (phosphatase and tensin homologue deleted on chromosome ten)/PI3 K (phosphatidylinositol 3-kinase)/Akt/mTOR (mammalian target of rapamycin) signaling pathway, which is commonly dysregulated in a broad array of human malignancies, controls the assembly of eukaryotic translation initiation factor 4F (eIF4F) complex through regulation of eIF4E binding proteins (4E-BPs) phosphorylation. And accumulated data over the past two decades implicated eIF4F complex as one of the promising targets for anticancer therapy. It has been confirmed that the translation initiation of mRNA coding for hypoxia-inducible factor-1α (HIF-1α) and survivin, which had been considered as the two major determinants of tumor radiosensitivity, are both controlled by eIF4F complex. Also, eIF4F complex controls the expression of VEGF and bFGF, the two well-known pro-angiogenic factors involved in developing radioresistance. Therefore eIF4F complex plays a pivotal role in regulation of radiosensitivity. In this article, we postulate that cell-permeable, phosphorylation-defective 4E-BP fusion proteins, which could be prepared by substituting the mTOR recognition motif located in N-terminal of 4E-BPs with protein transduction domain from HIV-1 TAT, HSV-1 VP22 or PTD4, could not only inhibit tumor growth but also enhance tumor response to radiation therapy through disruption of eIF4F complex assembly. In our opinion, the recombinant fusion proteins are superior to mTOR inhibitors for they do not cause immunosuppression, do not lead to Akt activation, and could be easily prepared by prokaryotic expression. If the hypothesis was proved to be practical, the cell-permeable, phosphorylation-defective 4E-BP fusion proteins would be widely used in clinical settings to improve tumor response to radiotherapy in the near future.

In addition to the typical size, Cryptococcus neoformans can enlarge its size to form titan cells during infection, and its diameter can reach up to 100 μm. Clinical reports about cryptococcal titan cells are rare. Most studies focus on aspects of animal models of infection with titan cells. Herein, we report the clinical and imaging characteristics and histopathologic features of 3 patients with titan cells and 27 patients with pathogens of typical size, and describe the morphological characteristics of titan cells in details. Histologically, 3 patients with titan cells show necrosis, fibrosis and macrophage accumulation. The titan cells appear in necrotic tissue and between macrophages, and have thick wall with unstained halo around them and diameters range from 20 to 80 μm with characteristic of narrow-necked single budding. There are also organisms with typical size. All 27 patients with normal pathogens show epithelioid granulomatous lesions. There is no significantly difference in clinical and imaging feature between the two groups. Cryptococcus neoformans exhibits a striking morphological change for the formation of titan cells during pulmonary infection, which will result in misdiagnosis and under diagnosis. The histopathological changes may be new manifestation, which need to be further confirmed by the study with animal models of infection and the observation of more clinical cases. Careful observation of the tissue sections is necessary.

As a common cause of chronic pelvic pain, endometriosis affects 10% of women of reproductive age. The most popular strategy for treating endometriosis is pseudo-pregnancy therapy. However, the efficacy of common systemic pseudo-pregnancy therapy is significantly attenuated by poor compliance because of so many side effects. While levonorgestrel-releasing intrauterine systems (LNG-IUS) successfully localize the effect of pseudo-pregnancy to the genital tract in treating endometriosis, it seemed to be insufficient to suppress the ovarian and extragenital endometriosis. We postulate that induction of a local pseudo-pregnancy via progestogen-loaded microsphere in the lesions of endometriosis may provide a more effective treatment with fewer side effects.

Bacterial vaginosis (BV) is one of the most common infectious diseases among sexually active women and is associated with the increased acquisition of a variety of sexually transmitted diseases. This study aimed to compare the efficacy of a non-antibiotic sucrose gel against an antibiotic metronidazole gel for the treatment of BV.A randomized, double-blinded, multi-center, parallel-group, placebo-controlled phase III clinical trial was conducted at eight hospitals in China. A total of 560 subjects with clinically diagnosed BV were randomly assigned into three groups for vaginally receiving sucrose, metronidazole, and placebo gels, respectively, twice daily for five consecutive days. The efficacy of therapeutic cure, defined as an achievement of both microbiologic cure (a Nugent score of 3 or less) and clinical cure (a resolution of the clinical findings from the baseline visit), was evaluated at the 1st and 2nd test-of-cure (TOC) visits at 7-10 and 21-35 days after the start of treatment, respectively.Therapeutic cure rates for sucrose, metronidazole, and placebo gel groups were 83.13%, 71.30% and 0.92%, at the 1st TOC, and 61.04%, 66.67% and 7.34%, at the 2nd TOC, respectively. While there was no significant difference between the sucrose and metronidazole gel groups at the 2nd TOC (P = 0.305), and sucrose gel was more effective than metronidazole gel at the 1st TOC (P = 0.009).These findings suggest that sucrose gel restores normal vaginal flora more rapidly than metronidazole gel and can be used as a novel treatment for BV.

Bit1 (Bcl2-inhibitor of transcription 1) is a mitochondrial protein that was found eliciting caspase-independent apoptosis when released into the cytoplasm, where it forms a complex with AES (amino-terminal enhancer of split). In our research, cytosolic fraction of suspended cultured human ovarian cancer cells was taken as immunogen, and recombined protein Bit1-His was used to select hybridomas. Two hybridoma cell lines secreting monoclonal antibodies (MAbs) against Bit1 were obtained by routine murine hybridoma technique. The MAbs were characterized by indirect ELISA, Western blotting, and immunohistochemistry, and it was found that they recognized distinct epitopes. The sandwich method to detect sera Bit1 was established, and a distinct difference of sera Bit1 between the ovarian carcinoma patients and normal controls was found. To conclude, these MAbs against human Bit1 may be useful for exploring the tumor apoptosis mechanism and monitoring patients' clinical data.

Objective GnRH antagonist cetrorelix could reserve the ovarian follicles during chemotherapy, but the mechanism remains unclear. The objectives of this study were to examine the overall effect of cetrorelix against ovarian failure and to define if the apoptotic process was involved. Methods Female SD rats were injected with cetrorelix before and after administration of saline, or cyclophosphamide (Cy), or oral etoposide (VP). Main outcome measures were the number of ovarian follicles, serum hormones, ovary histology and apoptotic markers. Results The females exposed to Cy or VP had reduced body and ovary weights, which could be restored by cetrorelix pretreatment. Single cetrorelix treatment could increase the number of primordial follicles, but reduce the number of growing and mature follicles. As a consequence, the ovaries exposed to cetrorelix prior to Cy or VP showed significantly higher numbers of follicles at all developing stages than those exposed to Cy or VP alone. Meanwhile, the ovarian apoptotic indexes as shown by TUNEL assay were reduced by cetrorelix pretreatment and the ovary expressed less caspases-3 and more Bcl-2 compared with chemotherapy alone. Moreover, the rats regained normal hormonal profile after cetrorelix pretreatment without any alterations in ovarian expression of estrogen receptor (ER)α, ERβ, or progesterone receptor (PR). Conclusion Cetrorelix could reduce the chemotherapy-induced ovarian damage through regulating the expression of Bcl-2 and caspases-3 in the ovary, without any expressional alterations of nuclear receptors, suggesting the apoptosis pathway involved.

OBJECTIVE The purpose of this study was to assess the usefulness of 3- and 4-dimensional ultrasonography (3D/4DUS) for the description of anomalies associated with trisomy 18 and to determine whether 3D/4DUS adds diagnostic information over what is provided by conventional 2-dimensional ultrasonography (2DUS) alone. METHODS Twenty-six fetuses subsequently proven to have trisomy 18 underwent prenatal ultrasonographic evaluations by both 2DUS and 3D/4DUS. Volume data sets were acquired by the same sonographers after the conventional 2DUS examinations and were explored with 4-dimensional imaging software by another independent examiner blinded to the indications of 2DUS. The findings detected by 2DUS and 3D/4DUS were compared with those acquired at autopsy. The sensitivity of each modality for detecting anomalies was calculated and compared by the McNemar test. RESULTS Excluding polyhydramnios, there were 131 anomalies confirmed postnatally in 26 fetuses with trisomy 18. There was a statistically significant difference in the sensitivity for detecting anomalies between 3D/4DUS and 2DUS (93.89% versus 73.28%; McNemar value = 23.31; P < .05), especially in anomalies of the face/neck (96.15% versus 65.38%; McNemar value = 6.13; P < .05) and extremities (96.3% versus 48.15%; McNemar value = 11.07; P < .05). Three- and 4-dimensional ultrasonography provided additional diagnostic information for 83.97% of the anomalies related to trisomy 18 and influenced the obstetric management of 4 fetuses. CONCLUSIONS Three- and 4-dimensional ultrasonography offered diagnostic advantages for many anomalies associated with trisomy 18, especially for anomalies of the extremities and face. This modality could be a powerful adjunct to 2DUS in the prenatal anatomic evaluation of fetuses with trisomy 18.

Objectives Exosomes, a type of membrane vesicles, released from tumor cells have been shown to be capable of transferring tumor antigens to dendritic cells and activating specific cytotoxic T-lymphocytes. Recent work has demonstrated the presence of high numbers of exosomes in malignant effusions. Umbilical cord blood (UCB) is a rich source of hematopoietic stem cells and from which a significant number of dendritic cells can be produced. We hypothesized that the exosomes released from metastatic ovarian carcinoma were able to present tumor specific antigen to dendritic cells derived from unrelated umbilical cord blood, then could stimulate resting T cells to differentiate and induce effective cytotoxicity. Study Design Exosomes were isolated by ultracentrifugation of malignant ascites from ovarian cancer patients (n = 10). Purified exosomes were further characterized by Western blot analyses and immunoelectronic microscopy. Dendritic cells were collected from unrelated umbilical cord blood and cultured in the presence of GM-CSF, IL-4 and TNF-α. Resting T cells were mixed with dentritic cells previously primed with exosomes and the cytotoxicity were measured by MTT method. T cells were activated by DCs presented with exosomes. Results 1) the exosomes isolated from the ascites were membrane vesicles of about 30-90nm in diameter; 2) the exosomes expressed MHC class I molecules, HSP70, HSP90, Her2/Neu, and Mart1; and 3)umbilical cord blood-derived DCs previously exosome-primed stimulated resting T cells to differentiate and produce effective cytotoxicity. Conclusions These results suggested that tumor-specific antigens present on exosomes can be presented by DCs derived from unrelated umbilical cord blood to induce tumor specific cytotoxicity and this may represent as a novel immunotherapy for ovarian cancer.

To construct the TRAIL gene eukaryotic expression vector modulated by the human telomerase reverse transcription gene core promoter and to evaluate the expression of the TRAIL gene in ovarian cancer cell line SKOV3 in vitro.The amplified TRAIL gene fragment was subsequently cloned into hTERT promoter-pIRES2-EGFP vector and CMVpromoter-pIRES2-EGFP vector. The hTERT promoter-pIRES2-EGFP-TRAIL and CMVpromoter-pIRES2-EGFP-TRAIL eukaryotic expression vectors were obtained, respectively. The plasmids were transfected into human ovarian carcinoma SKOV3 cell line by lipofeclin mediation and the positive clones were screened by G418. The expression of TRAIL gene was examined by RT-PCR, Western blot, immunocytochemistry and flow cytometry.All the constructed vectors were verified by enzyme digestion. The TRAIL gene of transfected cells was increased significantly (P0.01).An eukaryotic expression vector of the TRAIL gene modulated by the human telomerase reverse transcription gene core promoter has been constructed successfully and its steady expression in human ovarian cancer cell line SKOV3, which will be beneficial to further research into its role in regulating the biological behavior of ovarian cancer cells.

XIAP is one of the most important members of the inhibitors of apoptosis family. It is upregulated in various malignancies, including human ovarian carcinomas, it promotes invasion, metastasis, growth, and survival of malignant cells, and it also confers resistance to some chemotherapeutic drugs. We observed the effect of XIAP gene RNA interference (RNAi) on the proliferation, apoptosis, tumorigenicity, and chemosensitivity of the human ovarian carcinoma cells A2780/cp70.We used a human U6 promoter-driven DNA template approach to induce short hairpin RNA-triggered RNAi to block XIAP gene expression in the human ovarian cancer cell line A2780/cp70. A corresponding site-mutated vector was constructed as a negative control (pSilencer 2.1-NC). The expression of XIAP mRNA and protein among the stable transfected cells and the untransfected ones was detected by RT-PCR and Western blotting. The cell growth was examined and drug sensitivity was assayed using the MTT assay. Cell apoptosis was measured by flow cytometry and the tumorigenicity by using nude mice.Three stable transfected cell lines: A2780/cp70-s2, A2780/cp70-NC, and A2780/cp70-neo were established. The expression levels of XIAP gene mRNA and protein in A2780/cp70-s2 were 63.3% and 60.8%, lower than in A2780/cp70-NC, A2780/cp70-neo, and untransfected A2780/cp70 cells. The cell proliferation of A2780/cp70-s2 was reduced by up to 62.2+/-1.9%. Compared with the other cell lines, the changes in apoptotic rate in A2780/cp70-s2 was 31.1+/-1.3%, obviously increasing (P0.05). The knockdown of XIAP retards tumorigenicity in nude mice and after 21 days the average tumor weight of the A2780/cp70-s2 group was lower by 1.62+/-0.11g (P0.05). And the survival index in A2780/cp70-s2 cells was markedly reduced with the addition of 1 microM and 10 microM cisplatinum, respectively (P0.05).XIAP gene RNAi inhibited the proliferation of ovarian carcinoma cells and caused cells to be more sensitive to cisplatin through the reduction of its mRNA and protein. These results suggest that XIAP is an ovarian cancer-related gene and that XIAP is a potential target for therapeutic anti-cancer drugs.

Angiogenesis play a key roles in tumor growth, invasion and metastasis, and has become an attractive target for anticancer drug development. Though a number of anti-angiogenic agents had entered clinical trials, few of them could reproduce the spectacular results in cancer patients as that had been seen in pre-clinical tumor models. Therefore, exploring novel anti-angiogenic agents is highly deserved. SOX18, a member of the Sry-related HMG box-containing family of transcription factors, is expressed transiently in endothelial cells during the development of blood vessels. And mutations resulting in expression of dominant negative SOX18 have been shown to severely impair the vascular development. Recent research demonstrated that SOX18 is expressed during the initial steps of tumor vascularization and involved in regulation of the expression of the VEGF receptor Flk-1 and the vascular cell adhesion molecule-1 (VCAM-1). Moreover, allograft tumor growth in mice heterozygous for Ra(Op) (RaOp mice) which express a dominant negative mutant form of SOX18 (SOX18RaOp) that does not interact effectively with the endothelial partner protein MEF2C, was dramatically slower than that of wild-type mice. In this article, we postulate that recombinant cell-permeable dominant negative SOX18 mutants, prepared by fusion with protein transduction domains, would inhibit tumor angiogenesis with high efficiency by impairing endothelial tube formation. If the hypothesis was proved to be practical, the fusion proteins would show promise as single anti-angiogenic agents in cancer therapy.

To investigate the effect of hTERT promoter which regulated tumor targeting TRALL on the cervical cancer cell line HeLa.The mRNA expression of TRAIL on HeLa was examined by RT-PCR. The proliferation, apoptosis, motion of the transfected cervical cancer cell were detected by MTT, flow cytometry, and cell invasion assay respectively. The ultrastructure was observed by electron microscope.Compared with the control group, the expression of TRAILmRNA was significantly upregulated in hTERT-TRAIL (P0.05). The apotosis rate and the inhibitory rate of cell growth of hTERT-TRAIL was significantly high (P0.05). Electron microscope results indicated that hTERT promoter regulated tumor targeting TRALL facilitated the apoptosis of HeLa.The hTERT-TRAIL significantly inhibits the malignant proliferation and invasion ability of the cervical cancer cell line, and facilitates the apoptosisof HeLa, which lays a foundation for the treatment of patients with cervical cancer.

To construct an RNA interference vector to down-regulate X-linked inhibitor of apoptosis (XIAP) gene and study the RNA interference effect on the cell cycle and growth of ovarian cancer.Oligonucleotides of 64 base pairs for hairpin RNA targeting XIAP were designed, chemically synthesized, annealed, and cloned into the pSUPER vector. After identification by restriction digestion, the correct vectors were transiently transfected into SKOV3 cells, a human ovarian cancer cell line. The XIAP mRNA was detected by RT-PCR. The proteins were detected by western blot and indirect immunofluorescence staining. Flow cytometry (FCM) analysis and methyl thiazolyl tetrazolium (MTT) assay method were applied to measure cell cycle, cell growth and sensitiveness to cisplatin.SKOV3 cells had a high level expression of XIAP. The vector of RNA interference, which can interfere with XIAP gene was successfully constructed. After transient transfection, the expression of XIAP protein was significantly decreased in SKOV3 cells and the value of relative density was 3584 +/- 124, 2138 +/- 65, 1973 +/- 80 and 110 +/- 12, respectively (P = 0.0334). At the same time, the expression of XIAP mRNA was decreased accordingly and the value of relative density was 6674 +/- 274, 4532 +/- 107, 2322 +/- 57 and 1864 +/- 78, respectively (P = 0.0127). The FCM results showed that, the vector could increase the number of cells in G(1) phase compared with parent cells and compared with the cells transfected with pSUPER (P0.05); the number in S phase decreased compared with parent cells and compared with the cells transfected with pSUPER (P0.05). MTT results showed that pSUPER-siXIAP could inhibit the growth and proliferation of SKOV3 cells in a time-dependent manner, and inhibit the tumor cell growth.We successfully constructed an expressing hairpin RNA against the XIAP vector. The vector can effectively silence XIAP gene, increase the number of cells in G(1) phase, and slow down the growth of tumor cells. This may be a useful therapeutic strategy for XIAP over-expressing ovarian carcinomas.

To observe the effect of survivin gene repression by RNA interference (RNAi) on the radiosensitivity and the chemosensitivity to cisplatin of cervical cancer cell HeLa.The recombined eukaryotic expression plasmid pSilencer2.1-s2 containing human survivin gene small interference RNA (siRNA) was transfected into human cervical cancer cell line HeLa by using lipofectamine 2000. The expression of survivin gene mRNA and its protein was detected by semi-quantitative RT-PCR and western blot respectively. The changes of caspase-3 activity was assessed by kinase activity test. Cell apoptosis was examined by flow cytometry. The changes of cell radiosensitivity was observed by plate clone formation assay. Cell viability was examined by methyl thiazolyl tetrazolium (MTT) assay and 50% inhibitive concentration (IC(50)) of cisplatin was examined also.Compared with the cells which were transfected with negative control plasmid (HeLa-NC), pure plasmid (HeLa-U6 neo) and un-transfected cells (HeLa), the expression level of survivin gene mRNA and protein declined evidently in the cells transfected with pSilencer2.1-s2 plasmid (HeLa-s2), the expression inhibitory rates were (62.8 +/- 0.3)% and (60.1 +/- 0.5)%; the caspase-3 activity was enhanced and A(405) was 1.261 +/- 0.043 (P0.05); the apoptotic rate was increased obviously (29.23 +/- 1.41)% (P0.05). At the same dose of radiation, clone formation rate declined significantly (P0.05); at the same concentration of cisplatin, cell viability declined sharply and the IC(50) of cisplatin was (0.873 +/- 0.021) microg/ml (P0.05).Survivin gene repression by RNAi can enhance caspase-3 activity, induce cell apoptosis, significantly increase the radiosensitivity and chemosensitivity to cisplatin in human cervical cancer cell line HeLa.

To investigate the regulative effect of genistein on the apoptosis of the xenografted tumors of human ovarian carcinoma HO-8910PM cell on the nude mice.Human ovarian carcinoma HO-8910PM were cultured in vitro. The models of xenografted tumor were established by the transplantation of human ovarian carcinoma HO-8910PM on nude mice. The nude mice were randomly divided into four groups of three treatment groups (in which genistein were administered ip at 5, 25 and 50 mg x kg(-1) x d(-1), respectively, for 4 weeks) and one control group, the cell cycle and apoptosis of the xenografted tumors were measured by flow cytometry, the expression of bcl-2, Fas and FasL gene of xenografted tumors were determined by the immunohistochemistry and the morphology of tumor cells was observed by electron microscope.The tumor weights of 50 mg x xkg(-1) genistein group were more lighter (P0.05) compared to those of control group. The tumor cells in Go-G1 phase were increased, at saml time the cells in S-phase were decreased (P0.01) after the treatment of 50 mg x kg(-1) genistein. In addition the apoptosis rate of the cell treated with 50 mg x kg(-1) genistein group was (15.14 +/- 2.27)%, which was significantly higher than that in the control group (3.12 +/- 1.12)% (P0.01). The expression of bcl-2 of the xenografted tumors was decreased and the expression of Fas was increased in 50 mg x kg(-1) genistein group, both showing a significant difference from those in control group (P0.05). The apoptosis of the tumor cells were found more under electron microscopy in 50 mg x kg(-1) genistein group than those in control group.Genistein could significantly inhibite the proliferation and induce the apoptosis of the HO-8910PM cell of xenografted tumors by regulating the cell cycle and apoptoic gene in the nude mice.

Heparanase (Hpa) is an endoglycosidase that degrades heparin sulfate--the main polysaccharide constituent of extracellular matrix (ECM) and basement. It can enhance the invasive and metastatic potential of malignant tumors and cell lines by destroying ECM and basement. This study was to explore the effects of heparanase gene on the invasive ability of colorectal cancer cell line HT29.Heparanase gene was transfected into HT29 cells. Cell growth kinetics was assessed by MTT assay, and in vitro invasive ability was assessed with Boyden chamber. Xenograft and orthotopic implantation of histologically intact tumor in nude mice were constructed to observe the effect of exogenous expression of heparanase gene on invasive ability of HT29 cells.After transfection, the growth rate of heparanase-transfected HT29 (HT29-Hpa) cells was obviously higher than those of untransfected HT29 cells and empty vector-transfected HT29 (HT29-KZ) cells; the invasive cell number was significantly larger in HT29-Hpa cells than in untransfected HT29 cells and HT29-KZ cells (45.5+/-0.5 vs. 29.3+/-0.1 and 30.1+/-0.2, P0.01). The entity neoplasm formed by HT29-Hpa cells (12 mm x 9 mm x 10 mm) was bigger than that formed by untransfected HT29 cells (6 mm x 8 mm x 6 mm). The prevalence of liver metastasis caused by orthotopic implantation of xenograft was significantly higher in HT29-Hpa group than in control group (71.43% vs. 14.29%, P0.01).Exogenous heparanase gene can facilitate the growth, invasion, and metastasis of HT29 cells.

The aim of the present study was to determine molecular mechanism in hyperglycemia-induced congenital neural tube defects and the its potential pharmacologic rescuing agents. In order to explore these questions, six study groups of Sprague-Dawley rats were employed: Group 1 was normal control rats with normal diet; group 2 represented streptozotocin (STZ) -induced diabetic rats with congenital neural tube defects in offspring; group 3 included STZ-induced diabetic rats with normal offspring; groups 4,5 and 6 included rats exposed to the same STZ-induced diabetic condition, but receiving daily oral supplementation of 80 microg/mL of the sodium salt of arachidonic acid (AA), 400mg of vitamin E and a cocktail of a polyunsaturated fatty acid (safflower oil) plus an antioxidant ( vitamin E) respectively. Yolk sac cells were harvested at gestational day 12 from each rat group. Changes in MAPK signaling pathways were detected by western blot analysis using special antibodies directed against phosphorylated forms of extracellular signal regulated kinase (ERK), Jun N-terminal/stress-activated protein kinase (JNK/SAPK). Furthermore, activity of RAF-1, an upstream kinase in ERK1/2 signaling cascade, was evaluated by immunoprecipitation assay. The results showed that in yolk sac cells in embryopathic offspring from experimentally-induced diabetic rats, activities of ERK1/2 were dramatically decreased (group 2). Consisted with these observation, reduction in RAF-1 kinase activity could be discerned in these diabetic yolk sac cells. In contrast, activities of JNK1/2 were significantly increased in yolk sac cells of group 2. Under rescuing circumstance,activations of ERK1/2 and RAF-1 were increased, and JNK1/2 were decreased. MAP kinase signal pathway plays a very important role in hyperglycemia induced neural tube defects. The supplementation of polyunsaturated fatty acid arachidonic acid, and antioxidant vitamin E rescued conceptuses from diabetic embryopathy by triggering a restoration of normal membrane signaling pathways.

To investigate the expression of p27 protein and its significance in cervical carcinoma tissues.The p27 protein expression in 26 normal cervical tissues and 48 cervical carcinoma tissues was detected by streptavidin-peroxidase staining(SP method).The positive rate of p27 protein expression in 48 cervical carcinoma tissues was 39.5%, and that in 26 normal cervical tissues was 72.7%. The former was obviously lower than the latter(P0.01). In addition, The positive rate of p27 protein in 48 cervical carcinoma tissue was correlated with the differentiation degree of the carcinoma, clinical phase of the patients and lymph node metastasis. In the poorly-differentiated cancer cells, advanced phase patients and the patients with lymph node metastasis, positive rate of p27 protein expression was the lowest.The expression of p27 protein is related to differentiation degree of cancer cells, clinical phase of the patients and lymph node metastasis. Detection of p27 protein expression may be valuable to assess prognosis of the patients with cervical carcinoma.

Recent studies have shown that overexpression of Fas associated phosphatase-1 (FAP-1) can been detected in human ovarian cancer cell line SKOV3, suggesting that this overexpression may play an important role in the tumorigenesis and drug resistance of ovarian cancer. This study was designed to explore the effects of fas associated phosphatase-1 antisense oligonucleotide (FAP-1 ASODN) combined with carboplatin on the apoptosis of ovarian cancer cell SKOV3.Antisense oligonucleotide technique was used to transfect FAP-1 ASODN into SKOV3 cells. The expression levels of FAP-1 mRNA of SKOV3 cells with or without FAP-1 ASODN transfection were determined by reverse transcription- polymerase chain reaction (RT-PCR). The cell cycle and apoptotic rate were analyzed by flow cytometry (FCM). The growth and proliferation of SKOV3 cells were observed by MTT assay.After transfected FAP-1 ASODN for 24 hours, the expression of FAP-1 mRNA in SKOV3 cells was obviously reduced compared with those of the control and FAP-1 SODN transfection groups. When induced apoptosis with 40 microg/ml carboplatin for 24-72 hours, FCM results showed the apoptotic rate of "carboplatin+FAP-1 ASODN" group was higher than those of "carboplatin" group and "ASODN" group (P0.05), and the cell cycle was blocked in G1 phase. MTT results showed the cell inhibitory rate of "carboplatin+FAP-1 ASODN" group was 1.5-2 times those of "carboplatin" group and "ASODN" group (P0.05); but no significant difference was found between "carboplatin" group and "carboplatin+FAP-1 SODN" group (P0.05).FAP-1 ASODN transfection can suppress the expression of FAP-1 gene in SKOV3 cells and enhance the cell sensitivity to apoptosis induced by carboplatin, which implies that FAP-1 ASODN may reverse the drug resistance in ovarian cancer.

To explore the effect of insulin on apoptosis of cultured human trophoblast cells and its possible mechanism.Human trophoblast cells from early pregnancy women were cultured and divided into 3 groups; normal control group; H2O2, treatment group and insulin plus H2O2 treatment group. H2O2 was used to induce apoptosis of trophoblasts cells. Apoptotic rate was detected by flow cytometry. The effects of insulin on Bcl-2 expression and caspase-3 activity were also detected.H2O2 might induce apoptosis of trophoblast cells and typical morphological features of apoptotic cells was observed under electron microscope. Flow cytometry detection exhibited that insulin could reduce markedly H2O2-induced apoptotic rate of trophoblasts cells (P0. 01). Bcl-2 expression rate inH,O, treatment group was significantly lower than that in control group (P0. 01), while caspase-3 activity was distinctly higher than that in control group (P0. 01).Insulin could inhibit apoptosis of human trophoblasts cells induced by H2O2, which be may through decreased caspase-3 activity and increased Bcl-2 protein expression.

To explore the expressions of acetyl-heparanase mRNA, laminin ( LN) and laminin receptor ( LR) in 50 ovarian carcinoma, 33 ovarian carcinoma with lymph node metastasis, and 10 serous ovarian cystadenoma as well as their role in the metastasis of ovarian cancer.The transcription level of acetyl-heparanase mRNA, expressions of LN and LR were detected by in situ hibridization and immunohistochemical staining, respectively.The transcription level of acetyl-heparanase mRNA in ovarian carcinoma tissue and metastatic lymph nodes increased significantly, but its expression in primary focus was notably higher than that in metastatic lymph nodes (P0. 05 ). There was low expression of acetyl-heparanase mRNA in serous ovarian cystadenoma. The expression of acetyl-heparanase mRNA in malignant and benign tumor tissues had markedly difference (P0. 01 ). Expressions LN in both tissues mentioned above decreased while LR expression was high. The expression of acetyl-heparanase mRNA was negative correlation with that of LN, while positive with that of LR.The correlation among expressions of acetyl-heparanase mRNA, LN and LR suggests that heparanase is involved in the growth, invasion and metastasis of ovarian carcinoma.

Clinical studies showed there was no satisfying treatment of the patients with recurrent advanced ovarian cancer by using chemotherapy of combining platinum, and it is very important for oncologist to seek the second-line drugs to treat the patients with recurrent ovarian cancer. The current study was designed to evaluate the efficacy and toxicity of single agent Topotecan hydrochloride in the patients with recurrent advanced ovarian cancer.A total of 31 patients with recurrent ovarian cancer in a multiple centers clinical trial were treated with Topotecan 1.25 mg/m2 qd, i.v. inf. for 5 consecutive days every three weeks.Among 21 evaluable cases, overall response rate was 33.33%. Among 29 ITT population the response rate was 24.14%, including 3 CR and 4 PR. Main side effects were hematologic toxicities, 31.0% patients and 37.5% courses developed Grade III-IV leukocytopenia, 20.7% patients and 14.3% courses developed Grade III-IV thrombocytopemia. Non-hematologic toxicities were mild.Topotecan is effective for treating the patients with recurrent ovarian cancer who failed with platinum-based regimes.

Objective To analyze effects of high mobility group AT-hook 2 (HMGA2) on malignant degree, invasion, metastasis, proliferation and cellular morphology of ovarian cancer cells. Methods Three methods were applied to observe the effect on HMGA2 expression in ovarian cancer cells and ovarian epithelial cells. Results After the application of siRNA-HMGA2, number of T29A2-cell clones was decreased, there was significant difference compared with the negative control Block-iT. After application of let-7c, number of T29A2+ cell clones was decreased significantly, however, after the application of Anti-let-7, the number of clones restored, and there was no significant difference compared with the negative control group. After interference, the number of T29A2- cells which passed through Matrigel polycarbonate membrane were significantly lower than the negative control group. After the treatment of siRNA-HMGA2, let-7c and sh-HMGA2 respectively, growth and proliferation of T29A2-, T29A2+ and SKOV3 were slower, and the phenomenon was most obvious in SKOV3. Stable interference of HMGA2 induced mesenchymal-epithelial changes in the morphology of SKOV3-sh-HMGA2. Conclusions HMGA2 can promote malignant transformation of ovarian cancer cells, enhance cell invasion and metastasis, and promote cell growth and proliferation of ovarian cancer cells, which can cause ovarian cancer to progress rapidly and affect the quality of life.