Proteomic analysis of the influence of CO2 pneumoperitoneum in cervical cancer cells.

OBJECTIVE: The effect of CO2 pneumoperitoneum (CDP) on the oncology outcomes of patients undergoing laparoscopic radical hysterectomy for cervical cancer remains unclear. In this study, we investigated the effects of CDP on the proliferation of cervical cancer cells and examined the molecular mechanism. MATERIALS AND METHODS: We established an in vitro CDP model to study the effects of CDP on the proliferation of cervical cancer cells by Cell Counting Kit-8 (CCK-8) assay, xenografted tumor assay. Tandem mass tag-based quantitative proteomics were used to study the proteomic changes in HeLa cells after CDP treatment. Western blot assay was used to detect the expressions of PI3K/Akt signaling pathway proteins. RESULTS: CDP increased cell proliferation after a short period of inhibition in vitro and promoted tumorigenesis in vivo. Proteomic analysis showed that the expression levels of 177 and 309 proteins were changed significantly 24 and 48 h after CDP treatment, respectively. The acidification caused by CO2 inhibited the proliferation of cervical cancer cells by inhibiting the phosphorylation of PI3K and Akt. CONCLUSIONS: CDP promoted the proliferation of human cervical cancer cells after a short time of inhibition. The mechanism of which is related to the inhibition of phosphorylation of the PI3K/Akt signaling pathway.

Clinical application of the 2011 IFCPC colposcope terminology.

BACKGROUND: Colposcopy offers an accurate way to the diagnose of cervical precancerous lesions. However, the diagnostic accuracy of colposcopy is unsatisfied. This study was to evaluate colposcopic accuracy according to the 2011 International Federation of Cervical Pathology and Colposcopy (IFCPC) terminology. METHODS: A retrospective cohort study was performed in 1,838 patients who underwent colposcopy in Shandong Qianfoshan Hospital, Cheeloo College of Medicine, Shandong University from October 2013 to April 2018. Using conization or cervical biopsy pathology as the gold standard, the agreement between colposcopic diagnosis and pathologic diagnosis was calculated, and correlations between variables were analyzed. RESULTS: As an authoritative and widely used terminology for colposcopy diagnosis, the 2011 IFCPC terminology has certain clinical practicality and diagnostic accuracy. However, some signs such as mosaic, punctation, sharp border, inner border sign and ridge sign had high specificity but unsatisfactory sensitivity, which limited the diagnostic value. Therefore, we discussed the Lugol's staining, a very common sign in colposcopy, and analyzed the diagnostic significance of bright yellow staining in low-grade squamous intraepithelial lesion (LSIL) and mustard yellow staining in high-grade squamous intraepithelial lesion (HSIL). The results showed that mustard yellow may be a valuable indicator in the diagnosis of HSIL. CONCLUSION: The 2011 IFCPC colposcope terminology has standardized interpretations of the colposcopic findings and improved the accuracy of colposcopy diagnosis. The aceto-white epithelium still has important diagnostic value; however, the value of a few signs is needed to be discussed and new signs are expected to be discovered. Although the significance of Lugol's staining was diminishing, mustard yellow might be a valuable indicator for the diagnosis of HSIL.

Complications and feasibility analysis of ambulatory surgery for gynecological diseases in China.

ABSTRACT: The Chinese government is attaching great importance to the development of ambulatory surgery in order to optimize the healthcare system in China. The study aims to examine the complications and quality indicators of patients who underwent gynecological ambulatory surgery at a tertiary hospital in China.This was a retrospective study of patients who underwent ambulatory surgery between July and September 2019 at the Department of Gynecology of the First Affiliated Hospital of Shandong First Medical University. The patients were followed by phone at 30 days after discharge. The postoperative complications, mortality, unplanned re-operation, delayed discharge, unplanned re-hospitalization, and patient satisfaction were collected. The patients who underwent conventional hysteroscopic resection of uterine lesions during the same period were collected as controls for the economics analysis.A total of 392 patients who underwent ambulatory gynecological surgery were included. Fifteen patients had postoperative complications, and the total complication rate was 3.8% (15/392). Eight (8/392, 2.0%) patients had delayed discharge. There were no unplanned re-operations and deaths. There were two (2/392, 0.5%) cases of unplanned re-hospitalization. At 30 days after discharge, two patients were dissatisfied, and 390 cases were satisfied, for an overall satisfaction rate of 99.5%. Compared with conventional hysteroscopic resection of uterine lesions, ambulatory hysteroscopic surgery had a shorter hospital stay and lower total costs (P < .05) but similar surgery-related costs.Ambulatory gynecological surgery is feasible in China, with an acceptable complication profile and obvious economic and social benefits. Nevertheless, hospital management shall be reinforced.

Modified Lefort partial colpocleisis.

INTRODUCTION AND HYPOTHESIS: We present a surgical video that describes the technical considerations for performing a modified LeFort partial colpocleisis. METHODS: Hydro-dissection with diluted pituitrin was performed before the creation of anterior and posterior mid-line incisions through which lateral flaps were created bilaterally to expose the bladder and rectum fascia. Several purse-string sutures were placed to push the bladder and rectum back to their normal positions and reinforce the fascia under the vaginal wall. After removing the excess part of the vaginal wall, the lateral margins were re-approximated to create lateral channels that were wide enough to fit one finger. Perineoplasty was then performed to reduce the length of the genital hiatus. RESULTS: The procedure was performed in a 76-year-old woman with stage III vaginal vault prolapse (POP-Q C + 2), stage IV anterior prolapse (POP-Q Ba+5), stage II posterior prolapse (POP-Q Bp-1), and mild occult stress urinary incontinence. The patient recovered well postoperatively, without recurrent prolapse and/or stress incontinence during 6 months of follow-up. CONCLUSIONS: Our modified technique used traditional suture methods to strengthen the bladder and rectum fascia, keeping most of the vaginal wall to create a solid longitudinal septum in the center of the vagina that supported the vaginal vault.

Combination of metformin and RG7388 enhances inhibition of growth and induction of apoptosis of ovarian cancer cells through the PI3K/AKT/mTOR pathway.

Ovarian cancer is a gynecological cancer that has the highest mortality rate and is often resistant to conventional treatments. Therefore, development of new therapies is essential. Metformin (MET), which is the priority drug for treatment of type 2 diabetes, has received increasing attention because of its anti-tumor effects. Here, we examined combined anti-tumor effects of MET and RG7388, the only MDM2 (mouse double minute 2 homolog) antagonist that has entered phase III clinical trials, on ovarian cancer cell lines. We examined effects on proliferation by Cell Counting Kit-8 (CCK-8) and colony formation assays, and effects on apoptosis by flow cytometric analysis and Hoechst staining. Western blotting was used to measure protein expression in cells and tissues treated with MET and/or RG7388. Flow cytometry was used to measure reactive oxygen species (ROS). We also examined the effects of MET and/or RG7388 on inhibition of A2780 cell growth in vivo. The combination of MET and RG7388 significantly increased growth inhibition, apoptosis, and ROS of A2780 and SKOV3 cells compared with either agent alone. Additionally, in vitro and in vivo results showed that MET and/or RG7388 inhibited the phosphoinositide 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway and their combination had a stronger effect. Our findings suggest that the combination of MET and RG7388 enhances growth inhibition and apoptosis induction of ovarian cancer cells through the PI3K/AKT/mTOR pathway and accumulation of intracellular ROS.

Mutation of MDM2 gene in Chinese Han women with idiopathic premature ovarian insufficiency.

OBJECTIVE: Recent animal studies have demonstrated that the deletion of mouse double minute 2 (Mdm2) in mice leads to premature ovarian insufficiency (POI). The aim of the present study was to investigate whether mutations in the MDM2 gene contribute to POI in Chinese Han women. METHODS: The coding region of the MDM2 gene was examined in 54 Chinese Han women with idiopathic POI and 54 Han healthy controls. Two known single nucleotide polymorphisms (SNPs), rs937283 in 5'-UTR and rs2870820 in intron 1, were compared between both POI and control groups. RESULTS: There were no significant differences in the genotype distributions or allelic frequencies between the POI and control groups. No plausible causative mutations were identified. CONCLUSION: Our findings suggest that mutations in the coding region of the MDM2 gene may not represent a risk factor in the pathogenesis of idiopathic POI among Chinese Han women. Although we fail to confirm that MDM2 is a disease-causing gene, our study is the first to investigate the role of MDM2 in POI patients. Further studies with larger sample size from different ethnic populations are warranted.

Sclerosing stromal tumor of the ovary in a postmenopausal woman with estrogen excess: A case report.

RATIONALE: Sclerosing stromal tumor (SST) of the ovary is rare. We describe the first case of ovarian SST with estrogen excess with both clinical and serological evidence in a postmenopausal woman. PATIENT CONCERNS: A 70-year-old female who referred menopause 14 years ago was admitted with postmenopausal bleeding for 3 months. Ultrasonography revealed thickened endometrium of 6 mm and no adnexal masses. An elevated serum estradiol level of 49.78 ng/L was revealed in laboratory examination with normal ranges less than 27.25 ng/L in postmenopausal women. DIAGNOSES: The final histology diagnosis is SST of left ovary and endometrial hyperplasia with polyps. INTERVENTIONS: Laparoscopic hysterectomy and bilateral salpingo-oophorectomy were performed and a solid tumor with a diameter of 3 cm × 2 cm from the left ovary was found intraoperatively. OUTCOMES: Three days after removal of the tumor, the serum estrogen level was decreased to normal which indicated the estrogen activity of the tumor. LESSONS: To the best of our knowledge, it is the first reported case of ovarian SST with estrogen excess with both clinical and serological evidence. The present case illustrates the necessity to consider the rare possibility of ovarian SST as a cause for estrogen excess leading to postmenopausal bleeding. Hormonal evaluation (estrogens, androgens) should be considered in women with postmenopausal bleeding regardless of imaging examination.

miR-466 and NUS1 Regulate the AKT/Nuclear Factor kappa B (NFκB) Signaling Pathway in Intrauterine Adhesions in a Rat Model.

BACKGROUND Intrauterine adhesions (IUAs) are one of the most common reproductive system diseases in women worldwide. Emerging evidence has demonstrated that the upregulation or downregulation of genes plays an important role in IUAs. The aim of this study was to evaluate the role of NUS1 in IUAs in a rat model. MATERIAL AND METHODS The expression of miR-466 in intrauterine adhesions tissues was detected by using RT-qPCR assay. RT-qPCR, IHC, and Western blot were used to investigate mRNA and proteins expression, respectively, of NUS1. MTT and colony-formation assays were used to evaluate cell growth. Transwell assays were used to detect cell migration and invasion. To investigate miR-466 and NUS1 functions in vivo, we established a rat model. The level of epithelial-to-mesenchymal transition (EMT)-related markers was analyzed by Western blot assay. RESULTS NUS1 was upregulated in IUAs tissues, and the high expression level of NUS1 was positively correlated with the severity of IUAs. NUS1 promoted cell proliferation in vitro. NUS1 overexpression on cell migration and invasion promoted the EMT process in vitro and in vivo. NUS1 acted as a target of miR-466 and played the stimulative role by regulating AKT/NFkappaB pathway. CONCLUSIONS Our data suggest that miR-466 and NUS1 regulate proliferation and the EMT process through the AKT/NFkappaB pathway in IUAs in a rat model.

Laparoscopic hysterectomy in chronic renal failure patients with abnormal uterine bleeding.

BACKGROUND: Chronic renal failure (CRF) in women is frequently accompanied by endocrine disturbances leading to abnormal uterine bleeding (AUB). Recurrence of AUB is the most significant problem following current treatment approaches. Laparoscopic hysterectomy (LH) might be taken into consideration for patients who no longer want to preserve their fertility; however, these patients are in high-risk groups for minimally invasive surgery. Our goal was to determine whether LH is a good option for the treatment of AUB in patients with CRF when the associated risks are carefully managed. MATERIAL AND METHODS: This study included five patients who underwent LH for abnormal uterine bleeding between 2013 and 2016. Enhanced perioperative management and techniques were adopted to allow LH with minimized risks to the patient. RESULTS: All patients underwent LH successfully, without organ failure, heavy infection or abdominal cavity bleeding during the perioperative period. Operation times ranged from 95 to 152 min (mean time 117.6 min). The mean intraoperative blood loss was 24 ml. No recurrence was observed during the 18 months follow-up period. CONCLUSION: For CRF patients with AUB who no longer want to preserve their fertility, hysterectomy may be a better option, not only to permanently resolve the problem, but also to remove the risk of future uterine lesions. The improved protocols minimized the specific risks of laparoscopy arising from the patient's coexisting medical problems, making LH a feasible treatment approach in CRF patients with AUB.

Hyperamylasemia may indicate the presence of ovarian carcinoma: A case report.

RATIONALE: Reports of malignant ovarian tumor with hyperamylasemia are very rare. We present a patient with hyperamylasemia who was diagnosed with a malignant ovarian tumor. PATIENT CONCERNS: A 46-year-old woman was hospitalized complaining of a 2-day history of abdominal discomfort and fever. On physical examination, she showed abdominal distention and tenderness, with rebound pain. Laboratory evaluation showed significantly elevated serum amylase levels. Computed tomography (CT) revealed a solid mass with uneven density in the pelvis. DIAGNOSES: Based on her clinical symptoms and hyperamylasemia, she was suspected to have acute pancreatitis at first. However, the final pathology showed advanced serous papillary ovarian carcinoma. INTERVENTIONS: She underwent initial therapy for acute pancreatitis, followed by laparotomy once her symptoms had disappeared. A tumor mass with maximum diameter 12 cm was detected originating from the right ovary, and the patient underwent hysterectomy, bilateral salpingo-oophorectomy with omentectomy, and appendectomy. On the 14th day after the surgery, she received 5 courses of chemotherapy with paclitaxel and carboplatin. However, distant metastasis before the 6th course of chemotherapy were detected by CT, she was therefore changed to a chemotherapy regimen containing gemcitabine and capecitabine. OUTCOMES: The final pathology showed advanced serous papillary ovarian carcinoma. On the 14th day after the surgery, she received 5 courses of chemotherapy with paclitaxel and carboplatin. However, her serum CA125 levels rose again before the 6th course of chemotherapy, and CT of the abdomen and pelvis revealed multiple abnormal-density lesions in the peritoneum and pelvic cavity. We considered these to be metastases, and the patient was deemed unresponsive to her previous chemotherapy. She was therefore changed to a chemotherapy regimen containing gemcitabine and capecitabine, and remained on this regimen at the time of writing. LESSONS: Ovarian carcinoma should be considered as a possibility in patients with hyperamylasemia after ruling out other potential common causes. The final diagnosis depends mainly on the clinical manifestation, laboratory results, and CT examination, though pathology is mandatory to confirm the diagnosis. The main treatment is surgical excision.

Expression of Liver Receptor Homolog-1 (LRH-1) in Villi and Decidua of Patients with Unexplained Recurrent Spontaneous Abortion.

BACKGROUND In view of the important function of nuclear receptor liver receptor homolog 1 (LRH 1) in various biological processes and the physiological changes accompanying unexplained recurrent spontaneous abortion (USRA), our study was carried out to investigate the potential roles of LRH-1 in USRA. MATERIAL AND METHODS Thirty patients with URSA at early the early state of pregnancy were selected, and 30 patients with normal early pregnancy were also selected from Aug 2015 to Sep 2016 as a control group. The expression of LRH-1 protein in decidua and villi were detected by immunohistochemistry and Western blot analysis, and the expression of LRH-1 mRNA was detected by RT-PCR. The expression levels of CYP19 and P450scc were detected by RT-PCR and Western blot analysis at mRNA and protein levels, respectively. RESULTS The levels of LRH-1, CYP19, and P450scc mRNA and protein in villi of the patients in the URSA group were significantly lower than in the control group. There were no significant differences between the URSA group and control group in the levels of LRH-1, CYP19, and P450scc mRNA and protein in villi in decidua. CONCLUSIONS URSA was related to the reduced expression level of LRH-1 in villous tissues but not in decidua, and expression of LRH-1 may be related to the expression of CYP19 and P450scc. We believe that the expression level of LRH-1 can be used as a marker in the early diagnosis of URSA, and the regulation of LRH-1 expression many lead to new URSA treatments.

The role of peptidylarginine deiminase 4 in ovarian cancer cell tumorigenesis and invasion.

Peptidylarginine deiminase 4 (PADI4) is an enzyme that converts both histone arginine and mono-methyl arginine residues to citrulline, and it has been detected in various subtypes of ovarian cancer. However, the mechanism of action of PADI4 in ovarian carcinogenesis remains unknown. To examine the function of PADI4, we transfected two ovarian cancer cell lines, wild-type p53 A2780 and p53-null SKOV3, with PADI4-siRNA and negative control siRNA. The proliferation of both A2780 and SKOV3 cells decreased significantly following PADI4-siRNA treatment (P A2780 < 0.01; P SKOV3 < 0.001). The invasion and migration ability of A2780 cells also significantly decreased in response to PADI4-siRNA treatment (P < 0.001), but SKOV3 cells showed no such decrease. The apoptotic rate of A2780 cells increased in the presence of PADI4-siRNA, but there was no such increase in SKOV3 cells (P > 0.05). PCR arrays of A2780 cells treated with PADI4-siRNA revealed the up-regulated expression of six genes, including cell death-inducing DFFA-like effector a (CIDEA) and tumor necrosis factor receptor superfamily member 9 (TNFRSF9), and the down-regulation of seven genes, including integrin beta 3 (ITGB3) and BCL2-antagonist/killer 1 (BAK1). These results suggest an important role for PADI4 in the p53 pathway and the regulation of the proliferation, apoptosis, invasion and migration of ovarian cancer cells. Our study also demonstrated that PADI4 contributes to tumor metastasis by regulating the gene expression of insulin-like growth factor 1 (IGF1) and WAS/WASL-interacting protein family member 1 (WIPF1).

Overexpression of GOLPH3 protein is associated with worse prognosis in patients with epithelial ovarian cancer.

Golgi phosphoprotein 3 (GOLPH3) has important roles in the pathogenesis of cancer, and overexpression of GOLPH3 has been found in several kinds of cancers. However, the relationship between GOLPH3 overexpression and prognosis in patients with epithelial ovarian cancer remains unknown. This study aimed to investigate the relationship between GOLPH3 overexpression and overall survival in patients with epithelial ovarian cancer. The expression of GOLPH3 protein in tumor tissue was evaluated using immunohistochemistry. Seventy-five patients with epithelial ovarian cancer with the data of GOLPH3 expression and follow-up were included. GOLPH3 overexpression was significantly associated with advanced stage, histology, high grade, and lymph node metastases (P < 0.05). Kaplan-Meier analysis showed that patients with GOLPH3 overexpression had significantly poorer overall survival than those with low expression of GOLPH3 (log-rank P < 0.001). In the multivariate Cox proportional hazards analysis, GOLPH3 overexpression was independently associated with poorer overall survival (hazard ratio [HR] = 3.60; 95 % confidence interval (CI0 1.14-11.33, P = 0.03). Therefore, overexpression of GOLPH3 protein is closely related to poorer prognosis in patients with epithelial ovarian cancer.

High GOLPH3 expression is associated with a more aggressive behavior of epithelial ovarian carcinoma.

Overexpression of GOLPH3 (Golgi phosphoprotein 3, 34 kDa) is associated with the progression of many solid tumor types leading to an unfavorable clinical outcome. The present study examined the association between GOLPH3 expression and tumor development, clinicopathological factors, and prognosis of epithelial ovarian carcinoma. GOLPH3 expression was examined by immunohistochemistry in 18 normal ovarian samples, 28 benign tumors, 55 serous borderline ovarian tumors, and 135 epithelial ovarian carcinomas. The association of GOLPH3 expression with clinical characteristics, response to chemotherapy, and overall survival of epithelial ovarian carcinoma patients was analyzed on fresh tissue samples. GOLPH3 mRNA and protein expression in ovarian cancer and normal ovarian tissues were detected by real-time quantitative RT-PCR and Western blotting, respectively. The results are the following: (1) GOLPH3 immunostaining localized to the cytoplasm in two patterns, condensed into large granules with perinuclear distribution, and dispersed in the cytoplasm as fine granules. (2) GOLPH3 expression was higher in epithelial ovarian carcinoma than in normal ovarian tissues at the mRNA and protein level. The frequency of high-level expression of GOLPH3 increased progressively from benign (cystadenoma) to borderline neoplasms to malignant lesions. (3) Dispersed cytoplasmic GOLPH3 expression in epithelial ovarian carcinoma patients was highly correlated with FIGO stage (p < 0.001), tumor histological grade (p = 0.003), lymph node involvement (p = 0.001), and chemotherapy response (p = 0.034). (4) A dispersed pattern of GOLPH3 expression was an independent prognostic factor for poor overall survival. Patients with low dispersed cytoplasmic GOLPH3 expression had significantly longer overall survival than patients with high dispersed cytoplasmic expression. In contrast, GOLPH3 condensed expression was not correlated with clinicopathological features, chemotherapy response, or prognosis. GOLPH3 gene expression might play a role in tumorigenesis in epithelial ovarian carcinoma as upregulation of GOLPH3 expression is associated with a more aggressive tumor phenotype. GOLPH3 immunohistochemistry may be of value to predict the outcome of ovarian carcinoma patients.

Enforced effect of tk-MCP-1 fusion gene in ovarian cancer.

OBJECTIVE: The efficiency of HSV-tk/GCV system is not high because of insufficient gene transfer and incompletely initiative of host antineoplastic potency. The present study was designed to assess the antitumor efficacy of tk-MCP-1 on ovarian cancer in vitro and vivo. METHODS: A novel bicistronic expression system can help to improve the expression level of a gene in a stable manner. pLXSN/tk-MCP-1 co-expressing tk and MCP-1 genes was constructed using a pLXSN retroviral vector and an internal ribosome entry site sequence by restriction enzyme. Western blot was performed to determine tk and MCP-1 expression in the infected SKOV3. The GCV-sensitively tumoricidal activities of SKOV3/tk-MCP-1 with or without monocytes were compared to those of SKOV3 expressing HSV-tk or MCP-1. We investigated the growth of subcutaneous tumors in SCID mice immuno-reconstituted, and evaluated the antitumor effect of MCP-1 in conjunction with suicide gene. RESULTS: The significant GCV-sensitively tumoricidal activity of pLXSN/tk-MCP-1 was observed when compared with those of pLXSN/tk, pLXSN/MCP-1 and pLXSN/neo, especially when monocytes were added. The growth of subcutaneous tumors in SCID mice immuno-reconstituted was markedly suppressed by co-delivery of HSV-tk and MCP-1 genes, and the enhanced antitumor effect was associated with the recruitment of monocytes. CONCLUSION: These results demonstrated pLXSN/tk-MCP-1 presented an enhanced antitumor effects on ovarian cancer by orchestration of immune responses.