The feasibility of comprehensive staging surgery via laparoscopic operation in patients with ovarian cancer and its clinical significance / 肿瘤

Objective: To explore the feasibility and safety of comprehensive staging surgery via laparoscopic operation and its clinical significance. Methods: A retrospective analysis of medical records was conducted in 35 inpatients diagnosed of ovarian cancer (stage I, n = 29; stage II, n = 4; stage III, n = 2) in Department of Gynecology of Fourth Hospital of Hebei Medical University between January 2010 and January 2013. Of the 35 patients, 15 underwent comprehensive staging operation via laparoscopic operation and 20 via open operation. The general conditions during and after operation and the postoperative survival were observed and compared between the two groups. Results: All operations completed successfully in 15 patients in the laparoscopic operation group, and no perioperative complications were observed. The intraoperative blood loss and the rate of postoperative analgesia in the laparoscopic operation group were both less than those in the open operation group (P 0.05). There was also no significant difference in the cumulative survival rate between the two groups (P > 0.05). Conclusion: Comprehensive staging operation for ovarian cancer especially for early-stage cancer is feasible and safe. It has the merits of smaller wound, less blood loss during the operation and more rapid recovery. Copyright© 2014 by TUMOR.

Inhibitory effects of manumycin combined with cisplatin on the proliferation of human ovarian cancer cell line 3AO in vitro and in vivo / 肿瘤

Objective: To investigate the inhibition effects of manumycin combined with cisplatin (DDP) on the proliferation of human ovarian cancer cell line 3AO and the growth of transplantation tumors in nude mice, and to analyze its possible mechanisms. Methods: MTT colorimetric assay was performed to evaluate the cytotoxic effect of manumycin combined with DDP on 3AO cells. The model of nude mice bearing ovarian transplantation tumor was established. The expressions of survivin, NF-κB, vascular endothelial growth factor (VEGF) and caspase-3 proteins were detected by immunohistochemical method. The cell cycle distribution and apoptosis rate were examined by flow cytometry. Results: The proliferation of 3AO cells was inhibited by manumycin in a time- and dose-dependent manner (P <0.05). The manumycin of 10, 20, 40 or 60 μmol/L could strengthen the cytotoxic effect of DDP on 3AO cells. The tumor size in nude mice of each drug-treated group was significantly smaller than that of the control (untreated) group (P <0.05). Compared with the DDP alone and the manumycin alone group, the expressions of survivin, NF-κB and VEGF proteins were significantly down-regulated while the expression of caspase-3 protein was significantly up-regulated (P <0.05) in the combination (manumycin plus DDP) group (P <0.05). The apoptosis rates were increased gradually in the control group, manumycin group, DDP group and the combination group. In each drug-treated group, the number of cells in G0/ G 1 phase was decreased significantly, while which in G2/M phase was increased significantly (P <0.05), as compared with the control group. The number of cells in S phase did not show any changes in each group. Conclusion: The combination of manumycin and DDP shows an enhanced ability to inhibit the proliferation of human ovarian cancer cells 3AO and increase their apoptosis. This effect may be associated with the down-regulation of survivin, NF-

Effects of geldanamycin on cell proliferation and apoptosis of human ovarian cancer cell line SKOV3 and their mechanism / 肿瘤

Objective: To investigate the cytotoxic effects of geldanamycin on human ovarian cancer SKOV3 cells, and to explore the possible mechanisms of apoptosis and cell proliferation inhibition of SKOV3 cells induced by geldanamycin. Methods: MTT method was performed to evaluate the inhibitory effects of geldanamycin alone or in combination with cisplatin on proliferation of SKOV3 cells. Cell cycle and apoptosis rate of SKOV3 cells treated with geldanamycin were detected by flow cytometry. The expressions of Akt and Raf-1 proteins were examined by immunocytochemistry and flow cytometry. Results: The proliferation of SKOV3 cells was inhibited by different concentrations of geldanamycin in a time- and dose-dependent manners (P<0.05). Different concentrations of geldanamycin strengthened the inhibitory effect of cisplatin on proliferation of SKOV3 cells, and this inhibitory effect was strengthened with the increased concentration of geldanamycin within a certain concentration range. The apoptosis rate of SKOV3 cells was raised up gradually after treatment with 0-2000 nmol/L geldanamycin for 48 h, and the number of cells in G2/M phase was increased significantly (P<0.05). The results of immunocytochemistry and flow cytometry showed that the expressions of Akt and Raf-1 proteins in SKOV3 cells induced by geldanamycin were significantly down-regulated in a dose-dependent manner (P<0.05). Conclusion: Geldanamycin can inhibit the cell proliferation and induce the apoptosis of human ovarian cancer SKOV3 cells in vitro , which may be associated with downregulation of the expressions of Akt and Raf-1 proteins. Copyright© 2011 by TUMOR.

Inhibitory effects of enhanced expression of CD40L in ovarian cancer OVHM cells on the liver metastases in mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To examine the in vivo anti-metastatic effect of enhanced expression of CD40L cDNA in murine ovarian cancer OVHM cells (CD40L-OVHM) injected into the spleen on liver metastasis in mice.</p><p><b>METHODS</b>OVHM cells were inoculated into the spleen of 6 to 8 week-old female B6C3F1 (C57BL/6N x C3H/He) mice. The established liver metastasis was identified by histopathology (HE staining). OVHM cells, DNA-pMKITneo-OVHM cells or CD40L-OVHM cells were inoculated into the spleen of female B6C3F1 mice and the expressions of CD11c in splenic cells were detected by flow cytometry. The specific cytotoxicity of splenic cells was detected by MTT assay, and the serum cytokines of IFN-gamma, TNF-alpha, IL-12, IL-4 and IL-10 of the mice were measured by enzyme linked immunoabsorbent assay. The liver metastases and the survival time of the mice were also recorded.</p><p><b>RESULTS</b>The mouse models with liver metastasis by injecting tumor cells into the spleen of mice were established. The expression of CD11c and the specific killing rate in CD40L-OVHM cells group was significantly higher than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group. The expressions of IFN-gamma, TNF-alpha and IL-12 in the CD40L-OVHM cells group were much more increased than OVHM cells group and DNA-pMKITneo-OVHM cells group, but the expressions of IL-4 and IL-10 in the CD40L-OVHM cells group were decreased significantly (p < 0.05). The average weights of livers and spleens of mice in CD40L-OVHM cells group were significantly lower than those of DNA-pMKITneo-OVHM cells group and OVHM cells group. The survival time of mice in CD40L-OVHM cells group was also significantly longer than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group (P < 0.05).</p><p><b>CONCLUSION</b>The data directly demonstrate that the expression of CD40L in ovarian cancer cells (CD40L-OVHM) can enhance the proliferation and differentiation of dendritic cells in the spleen and induce specific cytotoxic effect of T cells in the spleen, and may regulate the immune function of peripheral blood cells and the immune balance between Th1 cells and Th2 cells, which maybe the possible mechanism induced by CD40L in mice inhibiting the development of liver metastasis.</p>

Enhanced expression of CD40L cDNA on ovarian cancer cell line OVHM induces the secretion of Th1 cytokines from dendritic cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To examine whether the enhanced expression of CD40L cDNA on murine ovarian cancer (OVHM) cells could induce the secretion of Th1 cytokines from dendritic cells (DC).</p><p><b>METHODS</b>OVHM cells were transfected with the full-length mouse CD40L cDNA by lipofectamine 2000 and then G418 resistant cells as positive cells were selected. They were examined for their expression of CD40L with flow cytometry. Bone marrow cells were firstly depleted of erythrocytes, macrophages, T and B cells with PE-conjugated magnetic beads, and then cultured in 10% FCS RPMI 1640 medium supplemented with recombinant mouse GM-CSF and IL-4 for 10 days. PKH67-labeled tumor cells were cultured with DC, and then the stained cells were analyzed for the expression of MHC-I, MHC-II, CD80, CD86, CCR7 in DC with flow cytometry. The expression of p40, p19, p35, p28, EBI3 subunits, IL-18, IFN-gamma, Mig gene in cocultured DC-tumor cells were detected by RT-PCR.</p><p><b>RESULTS</b>The CD40L cDNA was successfully transfected into OVHM cells. Bone marrow-derived DCs, when cultured with CD40L/OVHM, formed clusters with the tumors and showed an upregulated expression of MHC- I, MHC-II, CD80, CD86, CCR7. Expression of the IL-12, IL-23, IL-27, IL-18, interferon-gamma (IFN-gamma) and Mig (monokine induced by IFN-gamma) genes was induced in the DCs that were cultured with CD40L/OVHM but not with OVHM cells.</p><p><b>CONCLUSION</b>These data directly showed that the expression of CD40L on ovarian cancer cells facilitates the interaction between DCs and tumors, enhances the maturation of DCs, induces secretion of Th1 cytokines, especially for IL-12, IL-23 and IL-27, which maybe one of the possible antitumor mechanism for CD40L-transfected ovarian cancer cell line.</p>

Establishment of an in vivo model for duck hepatitis B virus infection using Hubei duckling / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To develop a standard duck hepatitis B virus (DHBV) animal model using a local Hubei species of duck, Ma Ya, and use it as an in vivo experimental system to study antiviral strategies against hepatitis B.</p><p><b>METHODS</b>Two-day-old Ma Ya ducklings were experimentally infected via intraperitoneal injection with the DHBV inocula which was collected from the transfected culture supernatant of 1.5-fold-overlength genome recombinant plasmid. Blood samples were taken twice or thrice a week during post-inoculation for 50 days. Viremia was quantified by serum real-time PCR to show the peak. Antiviral treatment of the DHBV-infected ducklings was started 3 d post-inoculation. The animals received oral administration of lamivudine (3TC) at a dose of 25 mg/kg/d for 5 d, followed by a maintenance therapy thrice weekly for 3 more weeks. Serum was quantified to show the viremia peak and liver biopsy specimens were analysed by Southern blotting and in-situ hybridization at the end of antiviral drug treatment.</p><p><b>RESULTS</b>The experimental infection rate of 2-day-old ducklings was 87.5%. Viremia started to be detectable on day 7 and reached a peak on day 11 post-inoculation, followed by a decrease and fluctuations. Four weeks of oral administration of 3TC led to a significant decrease in viremia peak during. This effect was not sustained, as a rebound in viremia was observed after drug withdrawal. Similarly, the analysis of liver biopsies at the end of 3TC treatment showed a marked decrease in DHBV DNA. However, after drug withdrawal a rebound of intrahepatic DHBV DNA was observed in duck livers.</p><p><b>CONCLUSION</b>The Hubei duck model with experimental DHBV infection of transfected supernatant is more suitable for the hepadnavirus biologic research due to its stability and practicability.</p>

The growth inhibition effects of TSLC1 gene on human hepatocyte carcinoma cell line HepG2 / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To study the effects of tumor suppressor in lung cancer-1 (TSLC1) on human hepatocyte carcinoma cell line HepG2.</p><p><b>METHODS</b>A full length of TSLC1 cDNA was amplified from RNA of normal human liver cells by RT-PCR, and it was cloned into a pCI-neo expression vector and transfected into human hepatocellular carcinoma cell line HepG2. The HepG2 cells transfected with this plasmid (experimental group) and those treated with pCI-neo vector (control group) and without any treatment (blank group) were compared. Cell morphology was studied microscopically and cell growth was analyzed with MTT assay. FACSort flow cytometry analysis was performed to assess the cell cycle distribution and apoptosis.</p><p><b>RESULTS</b>A stable cell line expressing TSLC1 protein was successfully established. Morphologically, cells of the experimental group were tightly aggregated when compared with those of the control and blank groups. The growth of TSLC1-transfected cells was significantly suppressed in vitro compared with those of the control and blank groups. The amount of G0-G1 cells was 63.66%+/-3.83% (P less than 0.01) in the experimental group, while those of the control and blank groups were 47.45%+/-0.91% and 54.47%+/-0.96% respectively. The amount of S phase cells in the experimental group, 22.90%+/-6.04%, was significantly lower (P less than 0.05) than that of the control group (36.58%+/-0.61%) and the blank group (33.61%+/-2.99%), which suggested a G0-G1 cell cycle arresting. The number of cells in early and late phase apoptosis (17.09%+/-0.20% and 16.11%+/-0.40% respectively) were significantly higher than those of the control and blank groups (P less than 0.01).</p><p><b>CONCLUSIONS</b>TSLC1 strongly inhibits the growth of HepG2 cells in vitro and induces apoptosis of the cells, suggesting that TSLC1 may have a tumor suppressor function in HCC.</p>

Antigenicity of major hydrophilic region II of hepatitis B virus surface antigen / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study whether the substitutions at the major hydrophilic region II (MHRII) of hepatitis B surface antigen (HBsAg) will impair the antigenicity of HBsAg.</p><p><b>METHODS</b>Four recombinant plasmids expressing mutant HBsAg (mtHBsAg) P1120T, C121S, K122I and T123N were constructed. HepG2 cells were transfected with the four plasmids and a plasmid expressing G145R HBsAg. The immunoreactivity of the cells expressing mtHBsAg with P1120T, C121S, K122I, T123N and G145R were detected by immunofluorescence (IF) staining and ELISA with 4 antibodies and 7 HBsAg diagnostic kits respectively.</p><p><b>RESULTS</b>mtHBsAg with P120T was recognized by mAb1 and mAb2. mtHBsAg with C121S and K122I was not recognized by any mAbs. mtHBsAg with T123N in lysates was recognized by mAb2, but not recognized in the supernatants.</p><p><b>CONCLUSION</b>Substitutions at amino acid positions 120-123 of HBsAg strongly impaired the antigenicity of HBsAg, a fact that was not appreciated previously.</p>

Inhibition of hepatitis B and duck hepatitis B virus replication by APOBEC3G / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3G (APOBEC3G) mediated antiviral activity against hepatitis B virus (HBV) and duck hepatitis B virus (DHBV).</p><p><b>METHODS</b>Total RNA was extracted from peripheral blood mononuclear cells (PBMCs), RT-PCR product was cloned into the EcoR I/Hind III restriction sites of the CMV-driven expression vector fused with a hemagglutinin fusion epitope tag at its carboxyl terminal. Replication competent 1.3 fold over-length HBV was constructed with full-length HBV of ayw subtype. The mammalian hepatoma cell HepG2 was cotransfected with the replication competent 1.3 fold over-length HBV and various amounts of CMV-driven expression vector encoding APOBEC3G-HA. Levels of HBsAg and HBeAg in the media of the transfected cells were determined by ELISA, HBV DNA. RNA from intracellular core particles was examined using Northern and Southern blot analyses. Chicken hepatoma cell LMH was cotransfected with head-to-tail dimer of an EcoR I monomer of DHBV and various amounts of CMV-driven expression vector encoding APOBEC3G-HA. DHBV DNA from intracellular core particles was examined using Southern blot analysis.</p><p><b>RESULTS</b>CMV-driven expression vector encoding APOBEC3G-HA and replication competent 1.3 fold over-length HBV were constructed. There was a dose dependent decrease in the levels of intracellular core-associated viral (HBV and DHBV) DNA and extracellular production of HBsAg and HBeAg. Levels of intracellular core-associated viral RNA were also decreased, but the expression of HBcAg remained almost unchanged.</p><p><b>CONCLUSION</b>APOBEC3G suppresses HBV and DHBV replication and also suppresses HBsAg and HBeAg expression.</p>