N-methyladenosine reader YTHDF2-mediated long noncoding RNA FENDRR degradation promotes cell proliferation in endometrioid endometrial carcinoma.

Dysregulation of long noncoding RNA (LncRNA) FENDDR has been shown to be closely related to the progression of several cancers. However, its role and upstream regulatory mechanism in endometrioid endometrial carcinoma (EEC) remains unclear. This study was conducted using the cancerous tissues of EEC patients (n = 60), EEC cell lines, and a xenograft mouse model. The expression level of LncRNA FENDRR was decreased and the N-methyladenosine (m6A) methylation levels of LncRNA FENDRR was elevated in cancerous tissues of EEC patients. In vitro experiments demonstrated that YTH domain-containing 2 (YTHDF2), an m6A reader, recognized the abundance of m6A-modified LncRNA FENDRR in EEC cells and promoted its degradation. LncRNA FENDRR overexpression suppressed cell proliferation and facilitated cell apoptosis in the EEC cell line HEC-1B by reducing the protein level of SRY-related HMG box transcription factor 4 (SOX4). Interference of LncRNA FENDRR reversed the inhibitory effect of sh-YTHDF2 on cell proliferation and the promoting effect of sh-YTHDF2 on cell apoptosis in HEC-1B cells by silencing FENDRR. Finally, in vivo experiments confirmed that overexpression of LncRNA FENDRR retarded the growth of EEC cells. In conclusion, YTHDF2-mediated LncRNA FENDRR degradation promotes cell proliferation by elevating SOX4 expression in EEC.

Value of serum and follicular fluid sirtuin (SIRT)1 and SIRT2 protein levels in predicting the outcome of assisted reproduction.

BACKGROUND: To explore whether serum and follicular fluid (FF), sirtuin 1 (SIRT1), and SIRT2 could predict the outcome of assisted reproduction. METHODS: All patients underwent in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) for the first time in the Reproductive Medicine Center of the First Affiliated Hospital of Zhejiang University Medical College from March 2018 to December 2018. According to cumulative clinical pregnancy outcomes, the patients were divided into a pregnancy group and non-pregnancy group. We measured the serum levels of SIRT1, SIRT2, anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) from the second to the fifth day of menstruation, and the levels of SIRT1 and SIRT2 in serum and FF on the day of human chorionic gonadotropin (HCG) injection and oocyte retrieval. RESULTS: A total of 125 patients met the inclusion criteria. The pregnancy group comprised 56 cases and non-pregnancy group 69 cases. There were significant differences in basal level SIRT2 (bSIRT2), AMH, antral follicle count (AFC), number of oocytes obtained, number of mature eggs, number of fertilized eggs, number of excellent embryos, number of blastocyst formations, and number of transferred high-quality embryos between the two groups. The area under the curve (AUC) values of bSIRT2, AFC, AMH, and age were significantly different from those under the opportunity reference line (P<0.05). In the subsequent correlation analysis, FFSIRT2, and HCG day serum SIRT2 were negatively correlated with age (r=-0.35, r=-0.19), and positively correlated with AFC (r=0.2, r=0.02). Serum SIRT1 on HCG day was negatively correlated with the number of blastocysts and the number of frozen embryos (r=-0.18, r=-0.21). Levels of FF SIRT1 and FF SIRT2 were significantly lower than those in serum SIRT1 and SIRT2, and there was no significant difference in serum SIRT1 and SIRT2 before and after ovulation promotion. CONCLUSIONS: The results suggest that bSIRT2 has significant statistical significance in predicting the cumulative number of pregnancies. When combined with AMH, AFC, and age, bSIRT2 can predict the cumulative pregnancy outcome. In addition, the level of serum SIRT1 and SIRT2 were not affected by ovulation promotion.

Comparative study of assisted reproductive outcomes between young patients with occult premature ovarian insufficiency and advanced-age patients.

OBJECTIVE: The purpose of this study was to compare the pregnancy outcomes among young patients with occult premature ovarian insufficiency (OPOI), advanced-age patients with diminished ovarian reserve (DOR), and advanced-age patients with normal ovarian reserve. METHODS: We retrospectively reviewed 324 women who underwent their first cycles of in vitro fertilization/intracytoplasmic sperm injection. The women were divided into the following groups: young women with OPOI, advanced-age women with DOR, and advanced-age women with normal ovarian reserve. The outcomes were compared among the different groups. RESULTS: The rates of live birth and embryo implantation in the young OPOI group were significantly higher than in the advanced-age DOR group, but comparable to those in the advanced-age normal ovarian reserve group. Moreover, the abortion rate was significantly lower in young OPOI patients compared with advanced-age patients with or without DOR. CONCLUSION: Higher embryo implantation and live birth rates and a lower abortion rate can be achieved in young patients with OPOI compared with older patients. The better outcomes in advanced-age patients with normal ovarian reserve compared with DOR may be related to egg quantity rather than quality.

Propofol promotes cell apoptosis via inhibiting HOTAIR mediated mTOR pathway in cervical cancer.

OBJECTIVES: Cervical cancer is one of the most common gynecologic malignant tumors. Propofol has been proposed to play a role of antitumor in various cancers. However, the functions and mechanisms of Propofol in cervical cancer is still not clear. METHODS: In vitro, the different concentrations of propofol were co-incubated with cervical cancer cell lines, including Hela, Caski and C-33A cells respectively. The pcDNA-HOTAIR plasmid was transfected into cells after the treatment of 10 µg/ml propofol. The cell viability and apoptosis were detected by MTT assay and TUNEL method. In vivo, propofol was injected into mice of transplantation tumor with Caski cells or with pcDNA-HOTAIR treated Caski cells. RESULTS: Propofol significantly decreased the cell viability and increased the cell apoptosis in Hela, Caski and C-33A cells, while HOTAIR overexpression promoted cell viability and inhibits cell apoptosis. mTOR/p70S6K protein expression levels were also markedly reduced by propofol but the effects were reversed with pcDNA-HOTAIR. In vivo, propofol inhibited the tumor size but had no inhibition effect in HOTAIR overexpression group. CONCLUSION: Propofol inhibited tumor size, cell viability and promoted cell apoptosis via inhibiting mTOR/p70S6K pathway mediated by HOTAIR in cervical cancer.

Fertility-sparing partial hysterectomy for gestational trophoblastic neoplasia: an analysis of 36 cases.

OBJECTIVE: To evaluate the success of partial hysterectomy in treating gestational trophoblastic neoplasia (GTN) and preserving fertility. STUDY DESIGN: A retrospective review was conducted on 36 patients with GTN who underwent adjuvant fertility-sparing partial hysterectomy at the Department of Gynecologic Oncology, Women's Hospital, School of Medicine, Zhejiang University, China, between 1991 and 2008. RESULTS: Of 36 patients, 34 had fertility-sparing partial hysterectomy; 2 required full hysterectomy due to excessive bleeding during surgery. All women went into remission from their disease. Twenty-five subsequently achieved clinical pregnancy, resulting in a total of 23 live births. CONCLUSION: Partial hysterectomy with chemotherapy can be effective in treating GTN while preserving future fertility. This should be considered as an option for women who wish to pursue pregnancy in the future.

[Family pedigree analysis in family member with history of hydatidiform mole and the NLRP7 gene expression].

OBJECTIVE: To evaluate the NLRP7 gene mutations and variants and their expression of genetic approach in hydatidiform mole patients with family history. METHODS: Six cases of mole patients with family members of mole history and 60 healthy women, taking blood, extracting DNA, the genetic mutation on NLRP7 screening and analysis, looking for mutations and corresponding amino acids, proteins control gene mutation found NLRP7 area. RESULTS: In 6 mole patients with family history: three patients were with sister's history of mole, and 2 of them familial recurrent hydatidiform mole (from family MoCh76 and family Ch77), there are 2 loci NLRP7 gene mutation. Screening patients from family MoCh76 for mutations in NLRP7 revealed in exon 3 and exon 5, amino acids [295G > T] and [1970A > T], proteins [Glu99X] and [Asp657Val], in a heterozygous. Screening patients from family Ch77 for mutations in NLRP7 revealed in exon 4 and exon 7, amino acids [1294C > T] and [2471 + 1G > A], proteins [Arg432X] and [Leu825X], in a heterozygous. Screening patients from family 105 for mutations in NLRP7 revealed no NLRP7 gene mutation. There were mother's history of mole in three patients, and they were not familial recurrent hydatidiform mole. Screening patients from family MoCh73 for mutations in NLRP7 revealed in exon 4, amino acids [1137G > C], proteins [Lys379Asn], in a heterozygous. Screening patients from family 106 and family 110 for mutations in NLRP7 revealed no NLRP7 gene mutation. There were not found mutations and variations in 60 cases of ethnic matched control group. CONCLUSION: NLRP7 mutations may be lead to familial recurrent hydatidiform mole.

[An outbreak of gastroenteritis caused by contaminated well water in a village, Henan province].

OBJECTIVE: To identify the cause and mode of transmission of a gastroenteritis outbreak in a village, Henan province. METHODS: Gastroenteritis patients were identified through family visits, interviewing the village doctors and reviewing diagnosis and prescription records at the village health clinic. Cases were defined as onset of one of the four symptoms from the village resident during July 20 to August 12, 2010. The symptoms would include diarrhea (≥ 3 times/day), abdominal pain, nausea or vomiting. A retrospective cohort study was conducted to assess the association between drinking raw well water or eating noodles rinsed by raw well water and gastroenteritis. Stools or vomits of the case-patients and the well water samples were tested for bacterial pathogens. RESULTS: Data for 60 case-patients were collected. All cases occurred in the northern part of the village. Persons who used water from a public well in the northern part of the village had an attack rate of 55%, which was 3.5 times of those who did not use the well water (16%) (RR = 3.5, 95%CI: 1.2 - 10). Results from the retrospective cohort study showed that drinking un-boiled water from the well was a risk factor (RR = 1.7, 95%CI: 1.3 - 2.3). Laboratory testing showed that total coliform and E. coli both greatly exceeded the limit considered safe for drinking, indicating there was fecal contamination in the well water. No bacterial pathogens were detected in the patients' stools or vomits. CONCLUSION: The outbreak was mainly caused by drinking contaminated water from the public well in the northern part of the village.

NLRP7 mutations in women with diploid androgenetic and triploid moles: a proposed mechanism for mole formation.

Hydatidiform mole is an aberrant pregnancy with abnormal embryonic development and hydropic placental villi. Common moles are sporadic, not recurrent and affect one in every 1500 pregnancies in Western societies. Approximately, half of common moles are complete and mostly diploid androgenetic, whereas the remaining are partial and mostly triploid diandric. NLRP7 has been found to be responsible for a recurrent form of molar pregnancies. Recently, we showed that patients with NLRP7 mutations have an impaired inflammatory response to various stimuli. To date, molar tissues analyzed from patients with NLRP7 mutations have been found to be diploid and biparental. In this study, we report 10 new non-synonymous variants and one stop codon found in patients and not in controls. We demonstrate the presence of different types of moles, diploid biparental, diploid androgenetic, triploid and tetraploid conceptions, in patients with NLRP7 variants. We document in vitro and in vivo early embryo cleavage abnormalities in three patients. We propose a two-hit mechanism at the origin of androgenetic moles. This mechanism consists of variable degrees of early embryo cleavage abnormalities leading to chaotic mosaic aneuploidies, with haploid, diploid, triploid and tetraploid blastomeres. Surviving embryonic cells that reach implantation are then subject to the maternal immune response. Because of the patients' impaired inflammatory response, androgenetic cells, which are complete allograft, are able to grow and proliferate. In women with normal immune system, chaotic mosaic aneuploidies may also occur during early cleavage, however, androgenetic cells would die after implantation or stay undetected, confined to a small portion of the placenta.

[Splenectomy in epithelial ovarian cancer: analysis of 32 cases].

OBJECTIVE: To investigate the clinicopathologic features, the complications of splenectomy and the survival of epithelial ovarian cancer patients with splenic metastasis. METHODS: A retrospective study was performed of 32 patients with epithelial ovarian cancer who underwent splenectomy for tumor cytoreduction at Zhejiang Cancer Hospital between Jan 1998 and Jun 2006. RESULTS: Of 32 patients, 23 patients (72%) were serous adenocarcinoma and 9 were non-serous adenocarcinoma. According to pathological grade, none was of G1, 11 were of G2, 21 were of G3. Postoperatively, 20 patients were left with no residual tumor, 7 were with < or = 2 cm and 5 were with > 2 cm residual tumor. Postoperative complications developed in 8 patients (25%), including subphrenic abscess, wound infection, gastric perforation, gastrorrhagia, phlebothrombosis, and bowel obstruction. The median follow up was 38 months, estimated 2-year and 5-year overall survival were 70% and 36%. Univariate analysis revealed that histological grade, residual tumor and courses of chemotherapy were influencing factors of the survival (P < 0.05), but multivariate analysis indicated that only residual tumor and courses of chemotherapy independently influenced survival (P < 0.05). CONCLUSIONS: In epithelial ovarian cancer patients with splenic metastasis, low grade serous adenocarcinoma is most common. Splenectomy as part of cytoreductive surgery is associated with modest morbidity and mortality. Residual tumor and courses of chemotherapy are independent factors associated with the prognosis of the patients.

[Oligonucleotide array of genomic expression in complete androgenic hydatidiform mole].

OBJECTIVE: To compare genomic expression differences between androgenic complete hydatidiform mole (AnCHM) and normal first trimester villi with similar gestation weeks, and search for potential adjuvant diagnostic molecular markers. METHODS: Short tandem repeat (STR) detection was used to identify AnCHM, human oligonucleotide array U133 Plus 2.0 was used to measure genomic expression differences between AnCHM and normal villi, and quantitative fluorescent RT-PCR was used to verify array of several genes. RESULTS: Nine of 11 histologically diagnosed complete hydatidiform moles were found to be AnCHM by means of STR, and the other 2 were biparental complete hydatidiform mole (BiCHM). Compared with villi, oligonucleotide array showed 279 genes (0.72%, 279/38 500) were over expressed and 1710 genes (4.44%, 1710/38,500) under expressed in AnCHM. Bioinformatics analysis found that differentially expressed genes were involved in multiple biological processes and pathways. Changes of imprinting genes, growth hormone genes and chorionic somatomammotropin hormone genes were especially remarkable. CONCLUSIONS: Pathogenesis of AnCHM is a complex process involving multiple genes and pathways. Altered expression of imprint genes may play important roles in the process.

[Study on parental origin of complete hydatidiform mole detected by polymorphisms of microsatellite].

OBJECTIVE: Through detection of 9 loci of polymorphisms of microsatellite to investigate the parental origin of complete hydatidiform mole. METHODS: Using the technology and method of multipolymerase chain reaction and electrophoresis in denatured polyacrylamide gel and silver stain detection, we carried out DNA analysis on 50 cases of complete hydatidiform mole diagnosed by histopathology and 50 copies of the couples' peripheral blood. RESULTS: There are 7 cases of biparental complete hydatidiform mole (14%); and 43 cases of androgenetic complete hydatidiform mole (86%) among 50 cases of complete hydatidiform mole. CONCLUSIONS: Detection of polymorphisms of microsatellite is a technology that can be used to identify the parental origin of complete hydatidiform mole. It is simple, quick, reliable and highly efficient.

[Effects of primary chemotherapy with single methotrexate on low-risk gestational trophoblastic neoplasia and influencing factors thereof].

OBJECTIVE: To evaluate the effects of primary chemotherapy with single-agent methotrexate (MTX) on low-risk gestational trophoblastic neoplasia (GTN) and the influencing factors thereof. METHODS: Sixty-one GTN patients with the score of < or = 6 according to the new International Federation of Gynecology and Obstetrics (FIGO) scoring system (2000) were divided into 2 groups: 51 patients were treated with single MTX 0.4 mg/kg daily for 5 days (MTX 5 d group), and 10 patients were treated with MTX on the days 1, 3, 5, and 7, and with folinic acid (FA) 0.1 mg/kg on the days 2, 4, 6, and 8 (MTX + FA group), both group with an interval of treatment course of 2 weeks. The serum level of human chorionic gonadotropin (hCG) was detected every week. If a plateau or increase of serum hCG appeared between 2 examination results, meaning tolerance to MTX, the patients concerned had to undergo different regimens of salvage chemotherapy, all with MTX as one of their components. Univariate and multivariate methods were used to analyze the relationships of different factors to the outcomes of chemotherapy. RESULTS: Thirty-five of the 51 patients of the MTX 5d group (68.6%) achieved complete primary remission, 3 of the 10 patients of the MTX + FA group achieved complete primary remission, and all patients achieved complete remission after salvage chemotherapy. Univariate analysis showed that the mean pretreatment serum level of hCG, duration between antecedent pregnancy and start of treatment, size of tumor, FIGO score, specific regimen of MTX were significantly associated with outcome of chemotherapy (P = 0.004, 0.022, 0.017, 0.005, 0.021 respectively). Logistic regression analysis showed that only three independent factors predictive for the outcome of chemotherapy: MTX regimen (OR = 2.476), FIGO score (OR = 1.431), and pretreatment hCG titer (OR = 1.001). CONCLUSION: Primary chemotherapy with single MTX regimen may still be one of the options for patients with low-risk GTN according to the new FIGO scoring system, though the rate of complete primary remission appears to be lower. All patients with low-risk GTN achieve complete remission after salvage chemotherapy. MTX regimen, FIGO score, and pretreatment hCG are independent risk factors of outcome of single MTX chemotherapy.

[Clinical analysis of 13 cases of gestational trophoblastic tumor misdiagnosed as ectopic pregnancy].

OBJECTIVE: To evaluate clinical-pathological features, diagnosis and therapy of gestational trophoblastic tumor (GTT) misdiagnosed as ectopic pregnancy. METHODS: From 1999 to 2003, a total of 13 patients with GTT misdiagnosed as ectopic pregnancy were retrospectively analyzed. RESULTS: The main symptoms were amenorrhea, abdominal pain, irregular vaginal bleeding. Serum beta-human chorionic gonadotrop in (hCG) was measured in 10 patients. Eight had hCG values above 10,000 IU/L; 3 had hCG values above 50,000 IU/L. The lesions of GTT misdiagnosed as ectopic pregnancy were fallopian tube, horn of uterus, peritoneal cavity, greater omentum, recto-uterine pouch. According to standards of the International Federation of Gynecology and Obstetrics (FIGO) the 13 patients were categorized as 6 of stage I, 2 of stage II, 3 of stage III and 5 of stage IV. Histologically they included 10 cases of choriocarcinoma and 3 of invasise mole. All patients were treated by complete surgical resection combined with subsequent adjuvant chemotherapy. CONCLUSIONS: Misdiagnosis leads to delay in therapy with resultant increased morbidity of GTT. Analysis on serial hCG is helpful to differential diagnosis between ectopic pregnancy and GTT.