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Objective To explore the folate receptor mediated (FRD) check and multi-function acetic acid white solution liquid based cervical cytology (TCT) application value in cervical cancer screening.Methods A total of 602 cases of patients was tested with FRD multi-function acetic acid white solution check,and TCT and cervical biopsy pathology examination.With the used of histopathological results as the gold standard,FRD multi-function acetic acid white analysis was compared with the TCT screening inspection results.Results For a total 602 patients with TCT screening,the positive rate was 21.8％ (131/602),including 36 cases of CIN Ⅰ level,41 cases of CIN Ⅱ level,24 cases of CIN Ⅲ level,and 30 cases of cervical invasive carcinoma.For the FRD multifunction white acetate solution screening,its positive rate was 23.8％ (143/602).No statistically significant difference was found between TCT and FRD screening (P ＞ 0.05).The missed diagnosis rate of FRD multi-function white acetate solution screening was 2.6％ in inflammation,and 21.1％ in cervical invasive cancer,and 3.8 ％ in CIN.The missed diagnosis rate of TCT screening was 7.2％ in inflammation,5.3％ in CIN Ⅰ,4.9％ in CIN Ⅱ,and 58.6％ in CIN Ⅲ]; whereas,its detection coincidence rate was 100％ in squamous cells carcinoma (SCC) and adenocarcinoma (AC).FRD multi-function acetic acid white solution screening had a sensitivity 80.92％,specificity 92.14％,positive predictive value 74.13％,and negative predictive value 95.59％.TCT examination had a sensitivity 90.84％,specificity 90.23％,positive predictive value 72.12％,and negative predictive value 97.25％.No significant difference was found between FRD and TCT methods (P ＞ 0.05).Conclusions FRD and TCT methods were both efficient in screening and evaluation for cervical lesions and cervical cancer.Because FRD method is limited in the deep tube for examination of cervical lesions; it cannot completely replace the TCT examination.However,FRD method is reliable,economic,and simple operation; it is suitable for primary hospitals census of cervical cancer
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<p><b>OBJECTIVE</b>To explore the effect of short peptides specifically binding to highly metastatic human ovarian cancer HO8910PM cells and their effect on the biological behavior of ovarian cancer cells.</p><p><b>METHODS</b>The phage-displayed peptide library was used to isolate the peptides binding and internalizing into the HO8910PM cells. Positive phage clones were characterized with DNA sequencing and bioinformatics analysis. The positive phage clones specifically bound to HO8910 cells were validated with immunofluorescence detection and enzyme-linked immunosorbent assay (ELISA). Furthermore, selected peptides were investigated for their cancer-related functions, including cell adhesion, spreading, motility, and invasion in vitro and in nude mice in vivo. The apoptotic index was detected by TUNEL assay, and VEGF expression by immunohistochemistry.</p><p><b>RESULTS</b>After 4 rounds of screening, apparent enrichment of phages was observed on the HO8910PM cells. ELISA assay showed that among the randomly selected 20 phage clones, 12 can specifically bind to HO8910PM cells. Immunofluorescence assay also showed that the selected positive phage clones can specifically bind to HO8910PM cells. The adherence test showed that the adherence rates of HO8910PM-peptide20, HO8910PM-peptide16 and HO8910PM cells were 49.0%, 96.8% and 100.0%, respectively. There was a significant difference between the cell adherence rates of HO8910PM-peptide20 and HO8910PM cells (P < 0.05). The peptide20 read as "THRVHLH" was a positive peptide and showed preferential binding to targeted cells. The peptide20 effectively inhibited tumor growth and metastasis in the nude mice, and the positive rates of VEGF protein in the tumor tissue of experimental, negative control and blank mice were 21.2%, 81.4% and 85.7%, respectively, showing that the positive rate of VEGF protein in the experimental group was significantly lower than that in the negative control and blank groups (P < 0.01), and the apoptotic index (AI) of the experimental group was (18.21 ± 2.49)%, significantly higher than the (3.76 ± 1.77)% in the negative control group and the (4.78 ± 1.57)% in the blank group (P < 0.01).</p><p><b>CONCLUSIONS</b>A novel short peptide able to specifically bind to highly metastatic human ovarian cancer cells is successfully screened. It can effectively inhibit the growth, invasion and metastasis of ovarian cancer cells, and provides an ideal vector in targeted drug therapy for ovarian cancer.</p>
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Animals , Female , Humans , Mice , Base Sequence , Cell Adhesion , Cell Line, Tumor , Cell Movement , Enzyme-Linked Immunosorbent Assay , Genetic Vectors , Immunohistochemistry , Mice, Nude , Molecular Targeted Therapy , Methods , Neoplasms, Glandular and Epithelial , Metabolism , Ovarian Neoplasms , Metabolism , Peptide Library , Peptides , MetabolismABSTRACT
Objective To investigate treatment effects of lentivirus mediated RhoA short hairpin RNA(shRNA) on xenograft tumor of ovarian cancer in nude mice in vivo and the underlying mechanism.Methods Human ovarian cancer cell line HO8910 were inoculated to establish subcutaneous xenograft model of human ovarian cancer.Tumor-bearing nude mice were assigned randomizely to three groups:LentiRhoA-sh group,Lenti-negative control (NC) group and phosphate buffered saline (PBS) group.lentivirus mediated RhoA shRNA,negative control lentivirus and PBS were respectively injected in the three groups.Effects of treatment were observed by tumor growth curve,tumor volume,tumor weight,and tumor inhibition rate.Xenograft tissues and liver,spleen,lung,and renal tissues were examined by hematoxylin and eosin (HE) staining or were detected by streptavidin-perosidase (SP)immunochemical method.The changes of RhoA gene expression in xenograft tissues after lentivirus mediated RhoA shRNA treated were also detected by real-time qPCR,immunochemistry and Western blot assay.Cell apoptosis in xenograft tissues were examined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL) method and apoptotic index (AI) were counted.Results Compared with Lenti-NC group and PBS group,the growth speed of xenograft in Lenti-RhoA-sh group delayed significantly after injection 9 days(P ＜ 0.01).Tumor volume (338 ± 114) mm3 decreased significantly in the Lenti-RhoA-sh group when compared with those in Lenti-NC group(1190 ± 332)mm3 and PBS group (1101 ± 396) mm3 (P ＜ 0.01).Tumor weight (0.23±0.11)g decreased significantly in the Lenti-RhoA-sh group when compared with Lenti-NC group (0.79 ± 0.19)g and PBS group (0.74 ± 0.17)g (P ＜ 0.01).Real-time qPCR result shown that the expression of RhoA mRNA (0.30 ± 0.05) decreased significantly in the Lenti-RhoA-sh group compared with Lenti-NC group (0.95 ±0.06) and PBS group(1.00 ±0.11 ; P ＜0.01).Western blot result showed that the expression level of RhoA protein decreased significantly in the Lenti-RhoA-sh group (0.14 ± 0.06) compared with those in Lenti-NC group(0.78 ± 0.14) and PBS group (0.75 ± 0.13;P ＜ 0.01).TUNEL staining displayed that AI significantly increased in the Lenti-RhoA-sh group (20.9 ± 3.4) ％ compared with those in Lenti-NC group(5.2 ±±2.0)％ and PBS group(6.0 ±2.1)％ (P ＜0.01).Conclusion Lentivirus mediated RhoA shRNA may be effectively down-regulate of the expression of RhoA,inhibit the growth of subcutaneous xenograft tumor of ovarian cancer in nude mice by increasing the cell apoptosis.
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BACKGROUND: It is difficult to in vitro isolate and culture the ovarian surface epithelium with high purity, strong vitality and stable biological characteristics. Tissue adherence and enzymatic digestion are commonly used for primary culture, but there are certain problems in cel col ection, cel viability and cel purity. OBJECTIVE: To establish a method for primary isolating, culturing and identifying human ovarian epithelium. METHODS: The ovarian surface epithelium was obtained with cel brush innovatively, and then the cells were isolated and purified with erythrocyte spal ation and differential adherence. The epidermal growth factor was added into the serum-free Dulbecco’s modified Eagle’s medium-F12 medium for cel culture. The cel morphology was observed under inverted microscope, and hematoxylin-eosin staining and immunocytochemical staining were used to identify the cells, then the growth curve was draw. RESULTS AND CONCLUSION: The ovarian surface epithelium became adherent after cultured for 24 hours, and reached fusion basical y after cultured for 6-12 days. The cells were polygonal or flat with strong transparency and refraction. The morphological characteristics of the cells were in line with those of the normal epithelial cells, and almost al the isolated cells could express the epithelial cells surface marker CK19. The cells could be passaged for 6-8 generations with wel growth and the cel growth curve was in “S” shape. The purity of the cells was more than 95%. The results suggest that cel brush method is simple to operate and can obtain a large amount of ovarian surface epithelium rapidly. The purity of the isolated cells can reach to 95% after treated with erythrocyte spal ation and differential adherence method and the cells are in stable growth.
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Objective To investigate the applied value of abnormalities of fetus with oligohydramnios in mid-pregnancy by anmioinfusion. Methods Fifty women with oligohydramnios diagnosed by ultrasonograph were analysed retrospectively. During the procedure 60-300 ml saline solution at 37 ℃ were infused into the amniotic cavity,made prenatal diagnosis at the same time necessarily. And examined the women to know whether the fetal anomaly existed or not after the amnioinfusion by the ultrasound. Results There were 15 cases with fetal anomaly, 8 cases with genitotirinary anomaly(2 cases associated with multiple organs anomaly), 1 case with single umbilical artery, 4 cases with chromosomal abnormality (2 cases associated with fetal growth restriction), 2 cases with pulmonary hypolasia. There was no significant difference in dilivery outcome between others and 35 cases with normal pregnant women (P >0.05). Conclusion Transabdominal amnioinfusion has double function on beth diagnosis and therapy for the oligohydramnios in the mid-pregnancy.
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Objective To evaluate the impact of uterine artery embolization (UAE) tor uterine fibroids on ovarian function in young woman. Methods From 50 young patients with symptomatic leiomyomas managed with UAE, completed follow-up 24-36 months of clinical symptomatic, ovarian function continue and pregnancy outcome. All patients participants ranged in age from 24 to 40 years, divided into lower 35 years group (30 cases)and over 35 years group (20 cases). Results There was no significant difference in serum FSH, LH and E2 levels between two groups before UAE and 3-6, 12, 24-36 months after UAE(P > 0.05).The comparison of serum FSH, LH and E2 levels between two groups at the same time points before and after UAE were no significant difference (P> 0.05). Menstrual blood loss reduced and duration of flow shorted after UAE in two groups, the differences were significant (P < 0.05). Cycle interval in over 35 years group was longer in 3-6 months after UAE than that before UAE, the difference was significant (P <0.05). There were 2 patients occurred transient secondary amenorrhea after UAE in lower 35 years group, 3 patients occurred in over 35 years group. One patient had menopausal syndrome in 37 months after UAE, and serum FSH and LH levels were found to heighten obviously, 47% pregnancies and 30% deliveries in lower 35 years group, 45% pregnancies and 20% deliveries in over 35 years group. Conclusion Transient secondary amenorrhea would have been occurred in young patients with uterine fibroid performed UAE, no prematureo varian failure and premature menopause.
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Objective To study the expression of epidermal growth factor receptor (EGFR) in cervical car-cinoma and cervical intraepithelial neoplasia(CIN),and to elucidate its relation with the genesis, infiltration, metas-tasis and prognosis of cervical carcinoma. Methods EGFR was determined by means of S-P immunohistochemistry in tissue of 100 cases of cervical carcinoma,60 cases of CIN and 40 cases of controls. Results The overexpression rates of EGFR were 0% (0/40), 51.67% (31/60),78.00% (78/100), respectively in normal cervical epithelium, CIN and cervical tumor tissues. The overexpression rate of EGFR was significantly higher in cervical tumor tissue than in control group(P<0.01). The overexpression of EGFR didn't demonstrate significant association with clinical staging, tumor size, pathological type, differentiation, cervical invasion depth, cervical canal invasion, lymphnode me-tastasis or the prognosis of cervical neoplasm (P>0.05). Conclusion Overexpression of EGFR is worsened with the severity of cervical lesion, suggesting that overexpression of EGFR is correlated with the genesis of cervical neo-plasms,which may be a valuable biological indicator of cervical carcinoma,but is not correlated with clinical patho-logical features and prognosis of cervical carcinoma.
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Objective To investigate effect of first, second, and third trimester placental factors (PF) on CD4, CCR5, and CXCR4 expression in human peripheral blood lymphocytes (PBLs), and to explore their influence on human immunodeficiency virus (HIV) vertical transmission.Methods Human peripheral blood mononuclear cells (PBMCs) were treated with first, second,and third trimester PF (concentration 25%) respectively for 24 hours. The expression of CD4, CCR5,and CXCR4 in PBLs, and the percentages of CCR5+, CXCR4+,and CCR5+CXCR4+ cells in peripheral blood CD4+ lymphocytes were determined with flow cytometry.Results All trimester PFs reduced CCR5 expression in PBLs. The efficiency of the first trimester PF was higher than that of the second and third trimester PF. The percentage of CCR5+ cells in peripheral blood CD4+ lymphocytes of PF groups was significantly lower than that of the control group, and the percentage of CCR5+ cells in peripheral blood CD4+ lymphocytes of the first trimester PF group was significantly lower than that of the second and third trimester group. The percentages of CCR5+CXCR4+ cells in peripheral blood CD4+ lymphocytes of PF groups were significantly decreased as compared with the control group, and the percentage of CCR5+CXCR4+ cells in peripheral blood CD4+ lymphocytes of the first trimester PF group was significantly lower than that of the third trimester PF group.Conclusion PF can reduce the expression of CCR5 in human PBLs and peripheral blood CD4+ lymphocytes, indicating that PF might reduce R5 virus infection via preventing HIV entry, and might play an important role in reducing R5 virus intrauterine infection.
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Objective To evaluate the clinical effect of ambulatory labor analgesia used in latent phase of the first stage of labor, which include labor progress, Apgar score after ambulatory labor analgesia begun to use when the cervix was different size dilatated. Methods Seventy-five parturient primiparas who had no complication were randomly divided into three groups: group Ⅰ: ambulatory labor analgesia was begun to use when the cervix was 1.0 cm dilated, group Ⅱ: ambulatory labor analgesia was begun to use when the cervix was 2-3 cm dilated, group Ⅲ: control group without use of ambulatory labor analgesia. Analgesic effects were observed, changes of uterine contraction were recorded by fetal monitor. Meanwhile, total stage of labor ,outcome of delivery and Apgar score were recorded. Results Duration of total stage of labor had no significant difference between group Ⅰ and group Ⅲ.The duration of the first labor stage was significantly longer in group Ⅰ than that in group Ⅱ(P
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Objective To study the effects of an extended-interval dosing regimen of gonadotropin releasing hormone agonist(GnRH-a) in the treatment of adenomyosis. Methods 20 patients with adenomyosis who asked for medicine treatment were divided into two groups randomly(n=10). The patients in the study group were given four doses of triptorelin depot 3. 75mg every 6 weeks in a new extended-interval dosing regimen. The other patients in the control group were given triptorelin 3.75mg in the conventional depot regimen of six doses,each given 4 weeks apart. The changes of degree of dysmenorrhea and the uterine volume were investigated. And the level of sexual hormone was also detected both before and after therapy.Results After six months of therapy,the reliving rate of dysmenorrheal was 100% both in the new regimen and the conventional regimen. The uterine volume was reduced at 46.7 % and 47. 5 % . And the level of LH, FSH and E2 were decreased significantly(P 0.05).Conclusion The use of the extended-interval dosing regimen of triptorelin depot in adenomyosis results in a consistent hypoestrogenised state,which is similar to that achieved by the conventional regimen. The new regimen thus reduces the cost of treatment obviously.