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OBJECTIVES@#Polycystic ovary syndrome (PCOS) is one of the most common endocrine diseases in women with reproductive age, which is associated with hyperandrogenism, insulin resistance, and ovulatory dysfunction. Progesterone receptor membrane component 1 (PGRMC1) can mediate progesterone to inhibit the apoptosis of ovarian granulosa cells and the growth of follicles, and to induce glucolipid metabolism disorder in ovarian granulosa cells, which is closely related to the occurrence and development of PCOS. This study aims to determine the expression of PGRMC1 in serum, ovarian tissue, ovarian granulosa cells, and follicular fluid in PCOS patients and non-PCOS patients, analyze the value of PGRMC1 in diagnosis and prognosis evaluation of PCOS, and investigate its molecular mechanism on ovarian granulosa cell apoptosis and glucolipid metabolism.@*METHODS@#A total of 123 patients were collected from the Department of Obstetrics and Gynecology in Guangdong Women and Children Hospital (hereinafter referred to as "our hospital") from August 2021 to March 2022 and divided into 3 groups: a PCOS pre-treatment group (n=42), a PCOS treatment group (n=36), and a control group (n=45). The level of PGRMC1 in serum was detected by enzyme linked immunosorbent assay (ELISA). The diagnostic and prognostic value of PGRMC1 was evaluated in patients with PCOS by receiver operating characteristic (ROC) curve. Sixty patients who underwent a laparoscopic surgery from the Department of Obstetrics and Gynecology in our hospital from January 2014 to December 2016 were collected and divided into a PCOS group and a control group (n=30). The expression and distribution of PGRMC1 protein in ovarian tissues were detected by immunohistochemical staining. Twenty-two patients were collected from Reproductive Medicine Center in our hospital from December 2020 to March 2021, and they divided into a PCOS group and a control group (n=11). ELISA was used to detect the level of PGRMC1 in follicular fluid; real-time RT-PCR was used to detect the expression level of PGRMC1 mRNA in ovarian granulosa cells. Human ovarian granular cell line KGN cells were divided into a scrambled group which was transfected with small interfering RNA (siRNA) without interference and a siPGRMC1 group which was transfected with specific siRNA targeting PGRMC1. The apoptotic rate of KGN cells was detected by flow cytometry. The mRNA expression levels of PGRMC1, insulin receptor (INSR), glucose transporter 4 (GLUT4), very low density lipoprotein receptor (VLDLR), and low density lipoprotein receptor (LDLR) were determined by real-time RT-PCR.@*RESULTS@#The serum level of PGRMC1 in the PCOS pre-treatment group was significantly higher than that in the control group (P<0.001), and the serum level of PGRMC1 in the PCOS treatment group was significantly lower than that in the PCOS pre-treatment group (P<0.001). The areas under curve (AUC) of PGRMC1 for the diagnosing and prognosis evaluation of PCOS were 0.923 and 0.893, respectively, and the cut-off values were 620.32 and 814.70 pg/mL, respectively. The positive staining was observed on both ovarian granulosa cells and ovarian stroma, which the staining was deepest in the ovarian granulosa cells. The average optical density of PGRMC1 in the PCOS group was significantly increased in ovarian tissue and ovarian granulosa cells than that in the control group (both P<0.05). Compared with the control group, the PGRMC1 expression levels in ovarian granulosa cells and follicular fluid in the PCOS group were significantly up-regulated (P<0.001 and P<0.01, respectively). Compared with the scrambled group, the apoptotic rate of ovarian granulosa cells was significantly increased in the siPGRMC1 group (P<0.01), the mRNA expression levels of PGRMC1 and INSR in the siPGRMC1 group were significantly down-regulated (P<0.001 and P<0.05, respectively), and the mRNA expression levels of GLUT4, VLDLR and LDLR were significantly up-regulated (all P<0.05).@*CONCLUSIONS@#Serum level of PGRMC1 is increased in PCOS patients, and decreased after standard treatment. PGRMC1 could be used as molecular marker for diagnosis and prognosis evaluation of PCOS. PGRMC1 mainly localizes in ovarian granulosa cells and might play a key role in regulating ovarian granulosa cell apoptosis and glycolipid metabolism.
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Criança , Gravidez , Humanos , Feminino , Síndrome do Ovário Policístico , Apoptose , Células da Granulosa , Metabolismo dos Lipídeos , Proteínas de Membrana , Receptores de ProgesteronaRESUMO
Objective To study the clinical and pathological characteristics of HPV-negative cervical cancer patients. Methods Retrospective analysis of 785 cervical cancer patients in Guangdong Women and Children Hospital from Jan. 2005 to Oct. 2015. By detecting high-risk HPV infection by flow-through hybridization genechip technique. Results (1) Among 785 cases of cervical cancer, 71 cases were negative for HPV infection tested by genechip technique, accounting for 9.0%(71/785), and the relative light units/cut off (RLU/CO) ratios of these 71 cases were less than 1 by hybird captureⅡ(HC-Ⅱ) methods. The results showed that the positive coincident rate of genechip technique detecting result with HC-Ⅱmethod was 100.0%(71/71). (2) There was no difference between 43 (60.6%) cases from 41-55 years old of 71 cases of HPV-negative patients and 392 (54.9%) cases from 41-55 years old of 714 cases of HPV infection patients (χ2=15.63, P=0.571). Among 71 cases of HPV-negative patients, 32 cases of patients with doing TCT, 6 (18.8%) cases for normal, 10 (31.2%) cases for atypical squamous cells of undetermined significance (ASCUS), 3 (9.4%) cases for atypical squamous cells cannot exclude high-grade squamous intraepithelial lesion (ASC-H), 3(9.4%) cases for low-grade squamous intraepithelial lesions (LSIL), 8 (25.0%) cases for hight-grade squamous intraepithelial lesions (HSIL), 2 (6.2%) cases for squamous cell carcinoma (SCC). And there were 391 cases of patients with doing thin-prep cytologic test (TCT) of 714 HPV infection patients, 60 (15.3%) cases for normal, 61 (15.6%) cases for ASCUS, 28 (7.2%) cases for ASC-H, 29 (7.4%) cases for LSIL, 164 (41.9%) cases for HSIL, 49 (12.5%) cases for SCC. There was no difference of TCT between HPV infection and HPV-negative patients (P>0.05). Among 70 cases from 71 patients with negative for HPV infection, there were 8(11.4%) cases in stage Ⅰa, 26 (37.1%) cases in stage Ⅰb1, 12 (17.1%) cases in stageⅠb2, 11 (15.7%) cases in stageⅡa, 10 (14.3%) cases in stageⅡb, 3 (4.3%) cases in stageⅢ-Ⅳ. There were 118(16.6%) cases in stageⅠa, 261 (36.8%) cases in stageⅠb1, 72 (10.1%) cases in stageⅠb2, 152 (21.4%) cases in stageⅡa, 87 (12.3%) cases in stageⅡb, 20(2.8%) cases in stageⅢ-Ⅳin 710 cases of HPV infection patients, in which there were no difference of clinical stage between HPV infection and HPV-negative patients (P>0.05). Among 69 cases from 71 patients HPV-negative infection, there were 51 (73.9%) cases for squamous carcinoma, 13 (18.8%) cases for adenocarcinoma, 5(7.2%) cases for adenosquamous carcinoma;and 593 (87.2%) cases for squamous carcinoma, 38 (5.6%) cases for adenocarcinoma, 39 (5.7%) cases for adenosquamous carcinoma, 10 (1.5%) case for others were in 680 patients of HPV infection, in which there was significant difference of adenocarcinoma between HPV infection and HPV-negative patients (χ2=11.96, P=0.001). Conclusions Flow-through hybridization genechip technique is the method of high sensitivity to detect high-risk type HPV, as like HC-Ⅱ method. HPV-negative of cervical cancer occurs mainly in 41-55 years old. Adenocarcinoma incidence is significantly higher in HPV-negative cases than those patients with infection of HPV positive.
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Objective To determine the clinical value of hysteroscopy technology in high risk planned abortion. Methods A retrospective study of clinical manifestation, medical tests, treatment methods and effects of 97 cases (hospitalized from June, 2013 to October 2014) was made in the research. Of the 97 cases, 53 underwent UAE, and 44 did not undergo UAE. The risk factors and treatment features were concluded. Results The hemorrhage risk relates to menopause period. The longer menopause period, the higher the hemorrhage risk will be. The hemorrhage risk has nothing to do with HCG. For those whose menopause is less than 56 days, hemorrhage risk showed no difference between groups with or without UAE. For those whose menopause is less than 56 days, hemorrhage risk decreases in group with UAE. Hemorrhage risk and hysterectomy possibility are higher if menopause has happened for more than 11 weeks. Conclusion Cesarean scar pregnancy needs to be detected and treated early, in this way, the possibility for complications can be lowered. For cases whose menopause has been more than e weeks, hemorrhage risk should be cautioned.
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Background and purpose: Resistance of cancer cells to chemotherapy is a major clinical obstacle to successful treatment and leads to poor prognosis for the patients. MicroRNA (miRNA) plays a vital role in tumor cells response to chemotherapeutic agents. This study plans to investigate the expressions of miRNA27a and miRNA451 in ovarian cancer and breast cancer cells and its correlation with drug resistance. Methods:A2780/Taxol cells were established using stepwise selection;Stem-loop quantitative real-time PCR (stem-loop RT-PCR) was used to detect expression of miRNA27a and miRNA451 in ovarian cancer and breast cancer cells. The A2780 and A2780/Taxol cells were transfected with the mimics or inhibitors of miRNA27a or negative control (NC) RNA and the mimics of miRNA451 or NC were transfected into MCF-7/ADM cells by LipofectamineTM 2000. The expression levels of MDR1 mRNA and P-glycoprotein (P-gp) were examined using RT-PCR and Western blot respectively. MTT method was used to analyze drug sensitivity. Results:The expression of miRNA27a was an average of (2.2±0.30) times higher in A2780/Taxol cells than in A2780 cells, with a significant difference between the two groups (P<0.05). The expression of miRNA451 was lower by 84%in MCF-7/ADM cells than in MCF-7 cells, with a signiifcant difference between the two groups (P<0.05). A2780/Taxol cells transfection with inhibitors of miRNA27a showed that the levels of MDR1 mRNA was decreased by (39±0.14)%, P-gp level [(26±5.3)%] decreased compared with the NC group [(43±6.7)%], the IC50 (0.53μmol/L) was less than the NC group (6.8μmol/L), and there was a signiifcant difference between two groups (P<0.05). Transfection of A2780 cells with mimics of miRNA27a led to increase MDR1 mRNA expression by (121±0.11)%and decrease the sensitivity to paclitaxel (IC50:0.2μmol/L vs 0.06μmol/L). There was a signiifcant difference between two groups (P<0.05). Transfection of MCF-7/ADM cells with mimics of miRNA451 showed that expression of MDR1 mRNA was decreased by (65±12)%, P-gp [(31±19)%] was less than the NC group [(83±12)%], the sensitivity of cells to adriamycin enhanced and the IC50 of adriamycin (4.61μmol/L) was less than the NC group (26μmol/L), and there was a signiifcant difference between two groups (P<0.05). Conclusion:The expressions of miRNA27a and miRNA451 are deregulated in A2780/Taxol and MCF-7/ADM cells respectively, which may play vital roles in drug resistance by regulating MDR1/P-gp expression directly or indirectly.
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Objective To evaluate the clinical significance of the detection of the Human Papilloma Virus (HPV) DNA typing in therapy of cervical diseases. Methods 780 cases have been studied. These cases which showed HPV-DNA positive and were diagnosed by biopsy as cervical cancer or cervical intraepithelial neoplasia( CIN Ⅰ-Ⅲ) were treated by operation or physical therapy. And 6 months and 12 months after the treatment,TCT and HPV DNA test were carried out. Results The prevalence of HPV-DNA types among the 780 cases descended from HPV 16 to 52,58,18,33 and 31. Detection rates of HPV-52 and 58 were highest among the patients with CIN Ⅰ. In patients with CIN Ⅱ,CIN Ⅲ,carcinoma in situ and invasive carcinoma,the positive rate of HPV-16 was obviously higher than other genotypes,and the difference was significant (P＜0. 01 ). 520 cases were followed-up after treatment,we found that HPV-DNA subsided within 3 months to 1 year in 432 cases. 88 cases still showed the HPV-DNA positive, among this group 48 cases were cytology diagnosed as normal or inflammation,but 14 cases were ASC-US,22 cases were LSIL and 4 cases were HSIL. Cytology abnormal cases were mere often detected in patients with persistent HPV-positive than in patients with HPV-negative. Conclusion Referring patients with cervical diseases the common HPV genotypes are 16,52,58,18,33 and 31. Especially HPV-16 are closely related with cervical cancer and high-level cervical intraepithelial neoplasis. HPV-DNA turn negative in most patients in 12 months after treatment. Persistent infection of HPV-DNA is related with the pathological changes persist.