Lentiviral vector-mediated short hairpin RNA targeting survivin inhibits abdominal growth of human endometrium xenograft in nude mice / 南方医科大学学报

<p><b>OBJEVTIVE</b>To investigate the inhibitory effect of lentiviral vector-mediated short hairpin RNA targeting survivin (LV-survivin shRNA) on the growth of human endometrium xenograft in the abdominal cavity of nude mice.</p><p><b>METHODS</b>The endometrium xenografts from 8 women with endometriosis were injected into the peritoneal cavities of 45 nude mice. The mice were then randomly assigned to receive intraperitoneal injection of LV-survivin shRNA, pGCL-NC-GFP (negative control) or PBS (blank control). Two weeks later, the number and morphometry of endometriotic lesions were quantified and the expression of survivin protein were detected by immunohistochemistry.</p><p><b>RESULTS</b>The formation of endometriotic lesions was significantly suppressed in mice receiving LV-survivin shRNA injection as compared with those in the two control groups (P/0.001). The mice in LV-survivin-shRNA group showed significantly down-regulated expression levels of survivin protein compared with those in the negative and blank control groups, presenting also necrosis in the endometriosis-like lesions in microscopic observation.</p><p><b>CONCLUSION</b>Lentiviral vector-mediated shRNA can effectively inhibit the expression of survivin in human endometrium xengrafts and suppress the formation and growth of endometriotic lesions in the abdominal cavities of nude mice.</p>

Expression of microRNA-100 and its correlation with drug resistance in human ovarian cancer SKOV3/DDP cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of microRNA-100(miR-100) and the relationship with cisplatin resistance in human ovarian epithelial cancer SKOV3/DDP cells.</p><p><b>METHODS</b>The SKOV3/DDP cells were transfected with the mimics or inhibitor of miR-100 or negative control RNA (NC) or inhibitor negative control RNA (inhibitor NC) by lipofectamine 2000. The experiment was divided into six groups: SKOV3 group, SKOV3/DDP group, miR-100 mimices group, NC group, miR-100 inhibitor group and inhibitor NC group. The expression of miR-100 and the cisplatin IC50 were measured by real-time PCR and CCK8 assay respectively.</p><p><b>RESULTS</b>(1)The cisplatin resistance index of SKOV3/DDP was 2.23; (2)The express level of miR-100 in SKOV3/DDP cells was significantly lower than that in SKOV3 cells (P<0.001); (3)After transfected with miR-100 mimics, SKOV3/DDP cells showed that the level of miR-100 was 38.29 times higher than that in the NC group(P<0.01). The cisplatin IC50 of miR-100 mimices group was significantly lower than that in the NC group (P<0.001); (4) After transfected with miR-100 inhibitor, the level of miR-100 0f SKOV3/DDP was decreased by 97.7%. The cisplatin IC50 of miR-100 inhibitor group was significantly increased as compared with that in the inhibitor NC group (P<0.001).</p><p><b>CONCLUSION</b>The expression of miR-100 is downregulated in SKOV3/DDP cells. Overexpressing miR-100 may effectively increase the sensitivity to cisplatin of human ovarian epithelial cancer SKOV3/DDP cells and may reverse cisplatin-resistance of EOC (epithelial ovarian cancer).</p>

Protective effect of lentivirus-mediated Bcl-2 gene transfection against phosphoramide mustard-induced apoptosis of human ovarian granulosa cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of lentivirus-mediated Bcl-2 gene transfection in protecting human primary ovarian granulosa cells against phosphoramide mustard (PM)-induced apoptosis.</p><p><b>METHODS</b>Granulosa cells were isolated from the follicle fluid of women undergoing in vitro fertilization and embryo transfer. The lentiviral vectors carrying Bcl-2 gene (pGC-FU-Bcl-2) and enhanced green fluorescence protein (pGC-FU-EGFP) were constructed and packaged into high-titer lentiviruses. The resulting recombinant lentivirus carrying Bcl-2 and EGFP genes or the empty vector were used to infect the primary human ovarian granulosa cells, followed by addition of PM in the cell culture, with untreated granulosa cells as the control. The cell apoptosis was detected by Annexin V and Hochst 33258 staining, and the expression of Bcl-2 protein was assessed using Western blotting.</p><p><b>RESULTS</b>The control granulosa cells showed an apoptotic rate of (1.93±0.28)%. The cells infected with pGC-FU-Bcl-2 prior to PM exposure had a apoptotic rate of (6.99±10.55)%, significantly higher than that of the control cells, but significantly lower than that of the cells with PM exposure only and those infected with the empty vector before PM exposure (P<0.05). The expression of Bcl-2 was the highest in the cells infected with pGC-FU-Bcl-2 prior to PM exposure (P<0.05).</p><p><b>CONCLUSION</b>Lentivirus-mediated Bcl-2 gene transfection can protect human ovarian granulosa cells against PM-induced apoptosis by upregulating Bcl-2 protein expression.</p>

Effect of lentiviral vector-mediated short hairpin RNA targeting survivin against endometriosis-like lesions in chick embryo chorioallantoic membrane / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of lentiviral vector-mediated short hairpin RNA targeting survivin (LV-survivin shRNA) on angiogenesis and growth of endometriosis-like lesions in chick en embryo chorioallantoic membrane.</p><p><b>METHODS</b>Eutopic endometrium from women with endometriosis was transplanted onto the non-vascular region of (CAM), where LV-survivin shRNA was delivered subsequently. The angiogenesis and the growth of endometriosis-like lesions in the CAM model were evaluated.</p><p><b>RESULTS</b>The angiogenesis and formation of endometriosis-like lesions were significantly suppressed in the CAM model by treatment with LV-survivin shRNA in comparison with those in the untreated CAM models and models treated with empty LV or DMEM (P<0.001). LV-survivin shRNA also caused a significantly higher cell apoptotic rate in the endometriosis-like lesions than the other treatments (P<0.001) and induced necrosis in the lesions.</p><p><b>CONCLUSION</b>LV-survivin shRNA can effectively inhibit angiogenesis induced by the eutopic endometrium and markedly suppress the formation of endometriosis-like lesions in the CAM model.</p>

Application of micro-computed tomography-based 3D visualization in diagnosis and treatment of cervical cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To define the optimal development time of micro-computed tomography (micro-CT) venography in cervical cancer patients and establish 3D CT-based digital pelvic model of the patients.</p><p><b>METHODS</b>Thirty patients with cervical cancer stratified by FIGO surgical staging underwent micro-CT scanning of the arterial phase and the venous phase with a delay time of 70, 90 and 120 s. The images were interpreted independently by two experienced radiologists to define the optimal development time in venous phase and establish the imaging diagnosis. Based on the pelvic CT scan data, we segmented the images using the abdominal medical image-3D visualization system followed by 3D image reconstruction to establish the 3D digital pelvic model using FreeForm Modeling System to modify the reconstructed images.</p><p><b>RESULTS</b>The optimal images were obtained by scanning with a 90-sec delay time. Micro-CT was not sensitive to IB1 phase or earlier phases (1/5), but efficient in advanced stages (≥IB2 phase). In our cases, 25 were diagnosed by micro-CT with a diagnostic accuracy of 64%. Based on these CT data, the pelvic 3D model covering the main organs, vessels, cervical neoplasm, and supplying vessels of the tumor in the pelvic cavity were successfully reconstructed to allow spatial observations of the uterus and the neighboring organs.</p><p><b>CONCLUSION</b>The 3D digital pelvic model reconstructed provides a means for staging cervical cancer and facilitates further surgical simulation studies.</p>

Effect of cervical conization through hysteroscopy for cervical intraepithelial neoplasia III / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the therapeutic effect of cervical conization through hysteroscopy in the treatment of cervical intraepithelial neoplasia (CIN) III.</p><p><b>METHODS</b>Seventy-four patients with CIN III underwent cervical conization through hysteroscopy (TCRC group), and 65 received cold knife conization (CKC group). The operating time, volume of blood loss, concordance rate with pathology, recurrence rate, rate of cervix adhesion and pregnancy rate were compared between the two groups.</p><p><b>RESULTS</b>The operating time, mean blood loss, cure rate, and recurrence rate were 15.1∓3.2 min, 12.5∓1.8 ml, 94.6%, and 5.4% in TCRC group, respectively, as compared with those of 25.8∓3.8 min, 21.6∓2.4 ml, 81.5%, and 18.5% in CKC group, all showing significant differences between the two groups (P<0.05).</p><p><b>CONCLUSION</b>Compared with CKC, TCRC has such advantages as less blood loss, shorter operating time, more accurate lesion localization, fewer complications, higher cure rate, and lower recurrence rate without significant adverse effect on pregnancy.</p>

CLINICAL ANALYSIS OF 12 CASES OF DISSEMINATED INTRAVASCULAR COAGULATION IN OBSTETRICS / 解放军医学杂志

To investigate manifestations and treatment of acute disseminated intravascular coagulation (DIC) in obstetrics, the medical records of 12 cases of acute DIC in obstetric patients admitted into our hospital from January 1993 to July 2001 were analyzed retrospectively. The result showed that the major causes of DIC in obstetric patients were amniotic fluid embolism and pregnancy induced hypertension. The main manifestations were bleeding, embolism or impairment of circulation. In all of the 12 patients accurate diagnosis was made, and treatment including elimination of causes, anti shock measures and prompt complement of coagulation factors. Nine patients received heparin early, 8 received antifibrinolytic agents rationally, and in 5 patients hysterectomy was performed. Nine patients were cured, 3 died. The key to decrease mortality of DIC in obstetric patients is to diagnose DIC early, to eliminate the causes promptly, to treat shock actively, and to use heparin and antifibrinolytic agents more rationally.