Effects of vitamin C, vitamin E, and molecular hydrogen on the placental function in trophoblast cells.

AIM: This study aimed to investigate the effects of three different antioxidants, namely vitamin C, vitamin E, and molecular hydrogen, on cytotrophoblasts in vitro. METHODS: Two trophoblast cell lines, JAR and JEG-3, were exposed to different concentrations of vitamin C (0, 25, 50, 100, 500, 1,000, 5,000 µmol/L), vitamin E (0, 25, 50, 100, 500, 1,000, 5,000 µmol/L), and molecular hydrogen (0, 25, 50, 100, 500 µmol/L) for 48 h. The cell viability was detected using the MTS assay. The secretion of human chorionic gonadotropin (hCG) and the tumor necrosis factor-α (TNF-α) were assessed and the expression of TNF-α mRNA was observed by real-time RT-PCR. RESULTS: Cell viability was significantly suppressed by 500 µmol/L vitamins C and E (P < 0.05), but not by 500 µmol/L molecular hydrogen (P > 0.05). The expression of TNF-α was increased by 100 µmol/L vitamin C and 50 µmol/L vitamins E, separately or combined (P < 0.05), but not by molecular hydrogen (0-500 µmol/L), as validated by real-time RT-PCR. But the secretion of hCG was both inhibited by 50-500 µmol/L molecular hydrogen and high levels of vitamin C and E, separately or combined. CONCLUSION: High levels of antioxidant vitamins C and E may have significant detrimental effects on placental function, as reflected by decreased cell viability and secretion of hCG; and placental immunity, as reflected by increased production of TNF-a. Meanwhile hydrogen showed no such effects on cell proliferation and TNF-α expression, but it could affect the level of hCG, indicating hydrogen as a potential candidate of antioxidant in the management of preeclampsia (PE) should be further studied.

Clinical observation of a modified surgical method: posterior vaginal mesh suspension of female rectocele with intractable constipation.

STUDY OBJECTIVE: To explore the feasibility and effectiveness of a modified posterior vaginal mesh suspension method in treating female rectocele with intractable constipation. DESIGN: Descriptive study (Canadian Task Force classification II-3). SETTING: The study was performed in the Study Center for Female Pelvic Dysfunction Disease, Department of Obstetrics and Gynecology, Tongji Hospital, Tongji University School of Medicine, Shanghai, China. The Study Center includes 15 physicians, most of whom have received advanced training in pelvic floor dysfunctional disease and can skillfully perform many types of operations in patients with such disease. Almost 1500 operations to treat pelvic floor dysfunctional disease are performed every year at the center. PATIENTS: Thirty-six women with rectocele with intractable constipation. INTERVENTION: Posterior vaginal mesh suspension. MEASUREMENTS AND MAIN RESULTS: All patients were followed up for 15 to 36 months. In 29 patients, the condition was cured completely; in 5 patients it had improved; and in 2 patients, the intervention had no effect. Insofar as recovery and improved results, the overall effectiveness rate was 94.4%. CONCLUSION: Posterior vaginal mesh suspension is an effective, harmless, and convenient method for treatment of female rectocele with intractable constipation. It has positive short-term curative effects, with few complications and sequelae. However, the long-term effects of posterior vaginal mesh suspension should be evaluated.

A 14-year multi-institutional collaborative study of Chinese pelvic floor surgical procedures related to pelvic organ prolapse.

BACKGROUND: It has been a global trend that increasing complications related to pelvic floor surgeries have been reported over time. The current study aimed to outline the development of Chinese pelvic floor surgeries related to pelvic organ prolapse (POP) over the past 14 years and investigate the potential influence of enhanced monitoring conducted by the Chinese Association of Urogynecology since 2011. METHODS: A total of 44,594 women with POP who underwent pelvic floor surgeries between October 1, 2004 and September 30, 2018 were included from 22 tertiary academic medical centers. The data were reported voluntarily and obtained from a database. We compared the proportion of each procedure in the 7 years before and 7 years after September 30, 2011. The data were analyzed by performing Z test (one-sided). RESULTS: The number of different procedures during October 1, 2011-September 30, 2018 was more than twice that during October 1, 2004-September 30, 2011. Regarding pelvic floor surgeries related to POP, the rate of synthetic mesh procedures increased from 38.1% (5298/13,906) during October 1, 2004-September 30, 2011 to 46.0% (14,107/30,688) during October 1, 2011-September 30, 2018, whereas the rate of non-mesh procedures decreased from 61.9% (8608/13,906) to 54.0% (16,581/30,688) (Z = 15.53, P < 0.001). Regarding synthetic mesh surgeries related to POP, the rates of transvaginal placement of surgical mesh (TVM) procedures decreased from 94.1% (4983/5298) to 82.2% (11,603/14,107) (Z = 20.79, P < 0.001), but the rate of laparoscopic sacrocolpopexy (LSC) procedures increased from 5.9% (315/5298) to 17.8% (2504/14,107). CONCLUSIONS: The rate of synthetic mesh procedures increased while that of non-mesh procedures decreased significantly. The rate of TVM procedures decreased while the rate of LSC procedures increased significantly. TRIAL REGISTRATION NUMBER: NCT03620565, https://register.clinicaltrials.gov.

Survivin small interfering RNA transfected with a microbubble and ultrasound exposure inducing apoptosis in ovarian carcinoma cells.

INTRODUCTION: Small interfering RNA (siRNA) has been used to knock down the expression of specific genes. However, its delivery remains a challenge. Recently, ultrasound combined with microbubbles has been used to deliver plasmid into cells and has shown much superiority. Whether a survivin siRNA transfected with a microbubble contrast agent combined with ultrasound exposure could inhibit survivin expression and induce ovarian cancer cell apoptosis was investigated. METHODS: The survivin gene was amplified and inserted into vector pEGFP-N1, resulting in psurvivin-EGFP. Three siRNA expression cassettes (SECs) targeting survivin were obtained by 2-step polymerase chain reaction. The SECs and psurvivin-EGFP were cotransfected into 293T cells. A functional SEC was selected and inserted into pMD18T, resulting in the survivin-targeting siRNA expression plasmid, which was transfected into SKOV-3 ovarian carcinoma cells using SonoVue, a microbubble contrast agent, and ultrasound exposure. Survivin expression was monitored by Western blot, and apoptosis was determined by fluorescence-activated cell sorting. RESULTS: One siRNA effectively inhibited survivin expression. SonoVue with ultrasound effectively delivered survivin siRNA to SKOV-3 cells, inhibited survivin expression, and induced apoptosis. CONCLUSIONS: Delivery of survivin siRNA using a microbubble contrast agent combined with ultrasound exposure can effectively inhibit survivin expression and induce apoptosis, providing a new promising approach for siRNA delivery in vivo.

Periostin Secreted by Carcinoma-Associated Fibroblasts Promotes Ovarian Cancer Cell Platinum Resistance Through the PI3K/Akt Signaling Pathway.

Periostin (POSTN) is a protein secreted by mesenchymal cells. Periostin is upregulated in several cancer types and overexpression is associated with poor prognosis. However, the functional role and molecular underpinnings of periostin in epithelial ovarian cancer (EOC) is unknown. In the present study, periostin was found to be significantly upregulated in EOC stroma. Functional studies revealed that periostin could decrease cisplatin (DDP)-induced apoptosis in EOC. Periostin led to DDP resistance in EOC cells, potentially through the PI3K/Akt signaling pathway. We generated periostin-overexpressing fibroblasts and found that EOC cells were resistant to DDP when co-cultured with periostin-overexpressing fibroblasts. The findings of the present study indicated that periostin secreted by cancer-associated stromal cells may be a potential therapeutic target for EOC.

Extensive profiling of circular RNAs and the potential regulatory role of circRNA-000284 in cell proliferation and invasion of cervical cancer via sponging miR-506.

Circular RNAs (circRNAs) are key regulators in the development and progression of human cancers, however its role in cervical cancer tumorigenesis is not well understood. The present study aims to investigate the expression profiles and potential modulation of circRNA on cervical cancer carcinogenesis. Human circRNA microarray was performed to screen for abnormally expressed circRNA in cervical cancer cells and circRNA-000284 was identified as one circRNA significantly upregulated in cervical cancer cells. Subsequent mechanistic investigations suggested that knockdown of circRNA-000284 suppressed cell proliferation and invasion, and caused G0/G1 phase cell cycle arrest. By performing anti-AGO2 RNA precipitation and luciferase reporter assay, we identified miR-506 as the circRNA-000284-associated miRNA. Furthermore, Snail-2 was identified as a direct target of miR-506, and circRNA-000284 could positively regulate the expression of Snail-2. Finally, the tumor promoting effect of circRNA-000284 was abolished by co-expression of miR-506 mimics or Snail-2 silencing vector. In conclusion, circRNA-000284 promotes cell proliferation and invasion in cervical cancer, and may serve as a promising therapeutic target for cervical cancer patients. Therefore, silence of circRNA-000284 could be a future direction to develop a novel treatment strategy.

Recipient bone marrow assimilates the myeloid/lymphoid reconstitution of distinct fetal hematopoietic stem cells.

The fetal liver (FL) is a source of hematopoietic stem and progenitor cells (HSPCs) for transplantation. However, whether FL-HSPCs collected at distinct developmental stages reconstitute similarly or differently in the recipient bone marrow (BM) remains undetermined. We examined this problem in a congeneic mouse transplantation model. We first analyzed the lineage components of FL from 12.5 days post-fertilization (dpf) to 18.5 dpf. The myeloid and lymphoid cells were dynamic in absolute number and ratio. The largest difference was between 12.5 and 16.5 dpf. The FL-HSPCs (Lin-CD150+CD48-) at these two time points were then respectively transplanted into the recipients. The difference in lineage reconstitution was undetectable at week 4 or 6 post-transplantation and afterward, indicating that the BM environment assimilated the transplanted cells. Profiling lineage-regulation genes of input and output HSPCs showed that the expression levels were much different in the former and almost the same in the engrafted HSPCs. Therefore, the recipient BM microenvironment could determine the developmental lineage-trends of FL-HSPCs.

Proportion of Uterine Malignant Tumors in Patients with Laparoscopic Myomectomy: A National Multicenter Study in China.

BACKGROUND: The Food and Drug Administration recently announced that the use of morcellation may cause fibroids or pelvic dissemination and metastasis of uterine sarcoma; therefore, the use of morcellation is limited in the USA. A large sample study is necessary to assess the proportion of uterine malignant tumors found in patients with laparoscopic myomectomy. METHODS: A national multicenter study was performed in China. From 2002 to 2014, 33,723 cases were retrospectively selected. We calculated the prevalence and recorded the clinical characteristics of the patients with malignancy after morcellation application. A total of 62 cases were finally pathologically confirmed as malignant postoperatively. Additionally, the medical records of the 62 patients were analyzed in details. RESULTS: The proportion of postoperative malignancy after morcellation application was 0.18% (62/33,723) for patients who underwent laparoscopic myomectomy. Nearly 62.9% (39/62) of patients had demonstrated blood flow signals in the uterine fibroids before surgery. And, 23 (37.1%) patients showed rapid growth at the final preoperative ultrasound. With respect to the pathological types, 38 (61.3%) patients had detectable endometrial stromal sarcoma, 13 (21.0%) had detectable uterine leiomyosarcoma, only 3 (3.2%) had detectable carcinosarcoma, and 5 (8.1%) patients with leiomyoma had an undetermined malignant potential. CONCLUSIONS: The proportion of malignancy is low after using morcellation in patients who undergo laparoscopic myomectomy. Patients with fast-growing uterine fibroids and abnormal ultrasonic tumor blood flow should be considered for malignant potential, and morcellation should be avoided.

Efficacy and safety of solifenacin succinate tablets versus solifenacin succinate tablets with local estrogen for the treatment of overactive bladder in postmenopausal women--a multicenter, randomized, open-label, controlled comparison study.

OBJECTIVE: The aim of the study was to compare the effectiveness and safety of solifenacin succinate tablets alone or combined with local estrogen for overactive bladder treatment in postmenopausal women. METHODS: This multicenter, randomized, open, parallel-controlled clinical trial enrolled 104 women between January 2012 and August 2013. Participants meeting the inclusion criteria were randomized 1:1 to 12 weeks of treatment with group A (solifenacin 5 mg qd + promestriene vaginal capsules intravaginally) or group B (solifenacin 5 mg qd). Before and after 12 weeks of treatment, symptoms (urinary urgency, frequency, and urge incontinence) were analyzed. Our primary outcome was the change from baseline to the end of treatment in the mean number of voids in 24 hours. Quality of life (QoL) was assessed using International Prostate Symptom Score and Overactive Bladder Symptom Score questionnaires and safety according to the incidence of adverse events. The t test or the Mann-Whitney U test was used to compare continuous variables, and the χ(2) test or Fisher's exact test was used to compare categorical variables. RESULTS: The median decreases in the mean number of voids in 24 hours in groups A and B were 5.2. and 4.3, respectively, which were not significantly different. The median decreases in urgency episodes in groups A and B were 2.0 and 2.5, respectively. In addition, the QoL scores significantly changed in both groups (both P < 0.05). The most common adverse event was dry mouth (19.2% in both groups). CONCLUSIONS: Solifenacin with or without local estrogen was effective and safe for overactive bladder treatment in postmenopausal women. The addition of local estrogen improved subjective feelings and QoL.

[Clinical research of a butterfly shaped mesh in the treatment of stress urinary incontinence].

OBJECTIVE: To investigate the effect of a butterfly shaped mesh in treatment of stress urinary incontinence (SUI). METHODS: From July 2003 to January 2004, 82 patients with SUI were treated with a butterfly shaped mesh. All patients were followed up at 1st, 3rd, 6th month after operation. According to their complaints, the cure standard of urinary incontinence is that the patient can control micturate by herself. The improved is that the times and volume of urinary incontinence is less than before. The inefficacy is that the patients' symptoms is not improved even more than before. RESULTS: All patients were operated under local anesthesia. Mean operation time was (27 +/- 9) min, and mean blood loss was (21 +/- 6) ml. Seventy two patients were able to micturate spontaneously at 2nd hour after operation and the volume of residual urine was less than 100 ml after 24 hour behind operation. They were discharged on the next day. 10 patients, an in-dwelling catheter had been used for 48 hours because the volume of residual urine was more than 100 ml. Seventy eight of 82 patients had no signs of stress incontinence. The remaining 4 patients with apoplexy history showed leakage of urine slightly. But their signs were improved obviously after a month. Seventy six patients had been followed up for from 1 to 6 month: 74 patients were completely cured, and the other 2 patients were improved. No urine retention, infection or bladder dysfunction were observed. CONCLUSION: A butterfly shaped mesh is effective, simple and safe procedure to treat stress urinary incontinence.

[Cloning, expression of the CP gene and antiserum preparation of zucchini yellow mosaic virus infecting Benincasa hispida Cogn. var. chieh-qua How].

A Guangzhou isolate of ZYMV infecting Benincasa hispida Cogn. var. chieh-qua How was identified by indicator tests and partial sequence amplification. The coat protein (CP) gene of this virus was amplified by RT-PCR, and ligated to the expression vector pET-22b(+). The recombinant plasmid pET-ZCP was transformed into E. coli BL21 (DE3) and then induced to express by IPTG. It was shown that the CP gene was highly expressed by SDS-PAGE and Western blot analysis. The molecular weight of the recombinant protein was about 33.0 kD. Antiserum with high specificity was produced after the rabbit was immunized with purified recombinant protein, and the titer was determined to be 1/4096 by antigen coating plate-ELISA (ACP-ELISA).

Pelvic reconstruction improves pelvic floor strength in pelvic organ prolapse patients.

OBJECTIVE: To evaluate pelvic floor muscle strength after the modified pelvic reconstruction procedure for pelvic organ prolapse (POP). MATERIALS AND METHODS: Patients were assigned to two groups consisting of 37 patients diagnosed with POP and undergoing modified pelvic reconstruction (reconstruction group), and 30 patients admitted to our hospital during the same period for other surgical indications (control group). Vaginal palpation of pelvic floor muscle strength was performed according to the modified Oxford grading system before operating on the two groups and again in the 3(rd) month following surgery for the reconstruction group. A comparative study was performed to evaluate the differences between the two groups and the improvement of pelvic floor muscle strength in the reconstruction group. RESULTS: The pelvic floor muscle strength was significantly improved postoperatively when compared with preoperative results in the reconstruction group (t = -17.478, p < 0.001). However, pre- and postoperative muscle strength in the reconstruction group was significantly lower relative to the control group, respectively (χ(2) = 63.293, p < 0.001; χ(2) = 31.550, p < 0.001). CONCLUSION: The modified pelvic reconstruction procedure could improve pelvic floor muscle strength in POP patients, which remains lower when compared with the normal population. Pelvic floor muscle strength should be included in the assessment of surgical outcomes in POP.

Laparoscopic repair of iatrogenic vesicovaginal and rectovaginal fistula.

OBJECTIVE: To investigate the clinical efficacy of laparoscopic repair of iatrogenic vesicovaginal fistulas (VVF) and rectovaginal fistulas. METHODS: Seventeen female patients with iatrogenic fistulas (11 cases of VVF and 6 cases of high rectovaginal fistulas) were included. All patients were hospitalized and underwent laparoscopic fistula repair in our hospital between 2008 and 2012. The mean age of the patients was 44.8 ± 9.1 years. The fistulas and scar tissue were completely excised by laparoscopy, orifices were tension-free closed using absorbable sutures, omental flaps were interposed between the vagina and the bladder or rectum, and drainage was kept after repair. RESULTS: Laparoscopic repair of fistulas was successful in all 17 patients. No complication was found during or after repair. No reoperation was needed after the repair. The operative time was 80.2 ± 30.0 minutes (range 50-140 minutes). The blood loss was 229.4 ± 101.6 ml (range 100-400 ml). The double J catheters were placed in 7 patients and removed 1-2 months after repair. Eight VVF patients underwent cystoscopy 3 months after laparoscopic repair and there were no abnormal findings. The follow-up time was 17.1 ± 6.5 months (range 8-29 months). CONCLUSION: Laparoscopic repair of VVF and rectovaginal fistulas is a safe and an effective minimally invasive procedure for treatment of iatrogenic fistula.

A new surgery for recurrent or persist stress urinary incontinence in females after primary mid-urethral slings.

OBJECTIVE: To retrospectively evaluate the efficacy of a new complementary mid-urethral sling surgery (Tong's hammock anterior, THA) in treatment of recurrent or persist stress urinary incontinence (SUI) in females after primary synthetic mid-urethral slings (MUSs). METHODS: THA was performed in 27 females with recurrent or persist SUI after primary MUSs from June 2005 and July 2010. These patients were followed up for one year, and clinical data including main complaints, operation duration, blood loss, efficacy and complications were reviewed. RESULTS: All 27 SUI patients were treated with THA surgery, a trans-vaginal mid-urethral sling on the descending pubic ramus. The average operation time was 39 min (range: 25-70 min), average blood loss was 70 ml (range: 20-120 ml). After urinary catheter removal, all patients could micturate and their average residual urine was 25.2 ml (range: 0-80 ml). The average hospital stay was 4.7 days (rage: 3-7 days). SUI symptom was persistent in 2 patients after THA surgery and the effective rate reached 92.5%. At 3 months, 6 months and 1 year after surgery, the effective rate was 92.5% (25/27), 92% (23/25) and 87.5% (21/24), respectively. 6 months after THA surgery, 2 were lost to follow up; 1 had recurrent SUI at 1 year and 1 had mesh erosion, 1 died of other diseases, and operative complications were absent after surgery. CONCLUSIONS: THA surgery is an effective method for treating recurrent or persistent SUI after primary MUSs. It is cheap, efficient, and easy to handle.

The comparison of an inexpensive-modified transobturator vaginal tape versus TVT-O procedure for the surgical treatment of female stress urinary incontinence.

OBJECTIVE: To compare the safety and efficacy of an inexpensive-modified transobturator vaginal tape procedure with the transobturator tension-free vaginal tape (TVT-O) procedure for the surgical treatment of female stress urinary incontinence (SUI). MATERIALS AND METHODS: Patients with SUI were randomly allocated to either the test group receiving the inexpensive-modified transobturator vaginal tape procedure or the control group receiving the GYNECARE TVT-O procedure. Treatment outcomes and Quality-of-life scores were recorded and analyzed between two groups. RESULTS: A total of 156 patients were enrolled in this trial. Eighty patients underwent the modified transobturator vaginal tape procedure. Among them 75(93.8%) were cured and 5(6.2%) were improved. The rest of the 76 patients underwent the GYNECARE TVT-O procedure with a 92% (70 of 76) cure rate and an 8% (6 of 76) improvement rate. No inefficient or aggravated cases occurred in both groups. The success rates between groups had no significant statistic difference (p > 0.05). The operative time, blood loss, hospital stay, and medical cost were significantly lower in the test group (p < 0.01); the increases in Quality-of-life scores were comparable between groups. CONCLUSIONS: The modified transobturator vaginal tape procedure is an efficacious and economic surgical treatment for female SUI.

HC-Pro protein of sugar cane mosaic virus interacts specifically with maize ferredoxin-5 in vitro and in planta.

Symptom development of a plant viral disease is a result of molecular interactions between the virus and its host plant; thus, the elucidation of specific interactions is a prerequisite to reveal the mechanism of viral pathogenesis. Here, we show that the chloroplast precursor of ferredoxin-5 (Fd V) from maize (Zea mays) interacts with the multifunctional HC-Pro protein of sugar cane mosaic virus (SCMV) in yeast, Nicotiana benthamiana cells and maize protoplasts. Our results demonstrate that the transit peptide rather than the mature protein of Fd V precursor could interact with both N-terminal (residues 1-100) and C-terminal (residues 301-460) fragments, but not the middle part (residues 101-300), of HC-Pro. In addition, SCMV HC-Pro interacted only with Fd V, and not with the other two photosynthetic ferredoxin isoproteins (Fd I and Fd II) from maize plants. SCMV infection significantly downregulated the level of Fd V mRNA in maize plants; however, no obvious changes were observed in levels of Fd I and Fd II mRNA. These results suggest that SCMV HC-Pro interacts specifically with maize Fd V and that this interaction may disturb the post-translational import of Fd V into maize bundle-sheath cell chloroplasts, which could lead to the perturbation of chloroplast structure and function.