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BACKGROUND:In vitro fertilization-embryo transfer is commonly used to solve infertility,but its success rate is not high,the more common reasons are poor endometrial receptivity,poor egg quality,etc.The follicular fluid melatonin can inhibit the aging of the ovary,to a certain extent,can promote the development of embryos,improve the probability of conception,but whether there is a correlation between the two is not known. OBJECTIVE:To explore the correlation between follicular fluid melatonin level and pregnancy rate of single-cycle in vitro fertilization-embryo transfer women. METHODS:A total of 112 female patients who received in vitro fertilization-embryo transfer treatment in the First Affiliated Hospital of Anhui Medical University from December 2020 to April 2021 were selected as the study subjects.They were divided into quartile array(Q1-Q5)according to the follicular fluid melatonin level from low to high.Among them,the melatonin level of group Q1 was<6.99 ng/L(n=18),that of group Q2 was 7.00-9.99 ng/L(n=26),that of group Q3 was 10.00-11.99 ng/L(n=27),and that of group Q4 was 12.00-13.99 ng/L(n=18);and melatonin levels in group Q5 were 14.00-19.99 ng/L(n=23).Clinical data characteristics of the five groups were compared.Multi-factor Logistic regression was used to analyze the correlation between follicular fluid melatonin level and pregnancy rate of women with single-cycle in vitro fertilization-embryo transfer and embryo transfer.A restricted cubic spline Logistic regression model was established to analyze the dose-response relationship,and the model was evaluated by clinical decision curve. RESULTS AND CONCLUSION:(1)Compared with the study population with the lowest melatonin quintile(Q1),with the increase of melatonin level(Q2-Q5),the levels of egg harvest and pregnancy success were gradually increased,and the body mass index was gradually decreased,and the differences were significant(P<0.05).(2)Multivariate Logistic regression analysis showed that after adjusting for confounding factors such as global mass index,number of eggs retrieved,luteinizing hormone,estradiol,progesterone and other confounding factors,follicular fluid melatonin level was still independently correlated with pregnancy rate of single-cycle in vitro fertilization-embryo transfer women(OR=1.538,95%CI:1.032-1.837,P<0.05),and there was significant difference in trend test of follicular fluid melatonin level from low to high quintile array(Ptrend<0.05).(3)The sensitivity test analysis showed that E value was 2.117.Subgroup analysis showed that the study population with higher levels of luteinizing hormone in follicular fluid had a more significant association between follicular fluid melatonin and pregnancy rate in single-cycle in vitro fertilization-embryo transfer women(P interaction=0.008).(4)The results of restricted cubic spline model analysis showed that there was a nonlinear dose-response relationship between follicular fluid melatonin level and pregnancy rate of single-cycle in vitro fertilization-embryo transfer women(P<0.05),and there was an overall positive correlation between follicular fluid melatonin level and pregnancy rate of single-cycle in vitro fertilization-embryo transfer women.(5)The results of clinical decision curve analysis showed that the follicular fluid melatonin level had important clinical value in predicting the pregnancy rate of single-cycle in vitro fertilization-embryo transfer women.(6)Follicular fluid melatonin level is closely related to the pregnancy rate of single-cycle in vitro fertilization-embryo transfer women,and with the decrease of follicular fluid melatonin level,the pregnancy rate of single-cycle in vitro fertilization-embryo transfer women also decreases.
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Objective To investigate the correlation between Yes-associated protein(YAP)nuclear expression and tumor size with prognosis of patients with epithelial ovarian cancer(EOC)and to study the role of YAP in EOC.Methods 120 patients with EOC were selected as the experimental group,including 38 patients with early stage(Ⅰ+Ⅱ)EOC and 8 2 patients with advanced stage(Ⅲ+Ⅳ)EOC.3 0 normal ovarian tissues obtained from patients with uterine leiomyoma were enrolled as the control group.Immunohistochemical(IHC)assay was em-ployed to determine YAP expression and sub-location.The relationship between YAP expression and the pathologi-cal parameters of the 120 patients with EOC was analyzed,so as to the prognosis of these patients.EOC cells(C13K and OV2008)were cultured with varying initial cell volumes.Ki67 expression and cell proliferation were tested by immunofluorescence and cloning assay respectively.YAP expression at mRNA and protein levels were de-tected by q-PCR and Western blot respectively when the cell conference of EOC cells reached to low(60％)and high(90％)cell density.Results The YAP nuclear expression was significantly higher in the EOC group com-pared to the control group(P＜0.05).The average diameter of stage Ⅰ+Ⅱ EOC was larger than that of stage Ⅲ+Ⅳ EOC(P＜0.01).The high nuclear expression of YAP was positively associated with pathological grade,clinical stage and the level of Ca125＞1 000 IU/ml,while negatively correlated with tumor size(all P＜0.05).Survival analyses showed that smaller tumor size(＜10 cm)and higher YAP nuclear expression were negatively as-sociated with the 3-year overall survival rate of EOC patients(P＜0.01).C13K and OV2008 cells cultured in the low density group exhibited a high number of clone formation,high Ki67 and YAP expression(P＜0.01).The down-regulation of YAP expression could decrease the cell viability of EOC cells in the low-and high-density groups(P＜0.05).Conclusion Higher level of YAP nuclear expression and smaller tumour size are inversely associated with the clinical prognosis of patients with EOC.Inhibiting YAP nuclear expression leads to a decrease in the prolif-eration capacity of EOC cells.
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Objective To investigate the application value of single nucleotide polymorphism(SNP)linkage analysis based on next-generation sequencing(NGS)technology in preimplantation genetic testing(PGT)of families with autosomal recessive polycystic kidney disease(ARPKD).Methods A family with ARPKD was selected,where the female member had a pregnancy ultrasound revealing polycystic kidney in the fetus.Genetic testing showed compound heterozygous mutations of the polycystic kidney/polycystic liver disease 1 gene(PKHD1),c.10444C>T(paternal)and c.4303del(maternal),with the c.4303del mutation being reported for the first time.Targeting the coding region of the PKHD1 gene,335 high-density tightly linked SNP sites were selected in the upstream and downstream 2M regions using multiplex polymerase chain reaction(PCR)and NGS.The couple′s SNP risk haplotypes carrying gene mutations were constructed.After in vitro fertilization,blastocyst culture was performed.Trophoblastic cells obtained from the biopsy were subjected to whole-genome amplification,and NGS was used for linkage analysis and low-depth chromosomal aneuploidy screening of the embryos.Sanger sequencing was used to verify the results of embryo linkage analysis.Results Among the 6 biopsied embryos,4 were mutation-free and euploid,1 exhibited heterozygous for the mutation and mosaic while another unstable sequencing data,making it impossible to judge.One of the mutation-free and developmentally healthy euploid embryos was implanted into the maternal uterus,resulting in the full-term delivery of a healthy baby.Conclusion Application of NGS-based SNP linkage analysis in PGT can effectively blocking the vertical transmission of ARPKD within families,while avoiding abortion issues caused by aneuploid embryos.This study is also the first PGT report target-ing the PKHD1 gene c.4303del mutation.
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Objective @#To investigate the factors influencing the pregnancy outcomes during frozen-thawed embryo transfer (FET) cycles in patients with polycystic ovary syndrome (PCOS) . @*Methods @#A retrospective analysis was conducted on patients ’data from 882 FET cycles . According to the pregnancy outcome , the patients were divided into non-implantation group (Group A) , abortion group ( Group B1) and live birth group ( Group B2) . Clinical data and laboratory parameters were compared among the three groups , and ordered Logistic regression analysis was used to study the factors influencing pregnancy outcomes after FET. Patients were also divided into four groups (C1-C4) based on the number of high-quality embryos obtained (0 - 3 , 4 - 6 , 7 - 10 , ≥11) , and their clinical data and laboratory parameters were compared .@*Results @#The clinical pregnancy rate , live birth rate , and miscarriage rate in the 882 treatment cycles were 71 . 09% (627/882) , 61 . 68% (544/882) , and 13 . 24% (83/627) ,respectively. Single-factor analysis showed significant differences in body mass index (BMI) , infertility type , human chorionic gonadotropin (hCG) day estradiol ( E2 ) level , number of retrieved oocytes , and number of high-quality embryos among Groups A , B1 , and B2 (P < 0. 05) . Further multiple Logistic regression analysis revealed that BMI(OR = 1 . 046 , 95% CI:1 . 001 - 1 . 093 , P = 0. 044) and a history of previous pregnancy(OR = 1 . 417 , 95% CI:1 . 030 - 1 . 950 , P = 0. 032) were independent risk factors for successful FET in PCOS patients , while an increased number of high-quality embryos was an independent protective factor for successful pregnancy. Based on the results of Group B2 , compared to Group A , OR = 0. 920 , 95% CI:0. 880 - 0. 962 , P = 0. 000;compared to Group B1 , OR = 0. 923 , 95% CI:0. 862 - 0. 988 , P = 0. 022 . Compared with the other three groups( C1-C3) , the total amount of gonadotropin (Gn) in the C4 group was the lowest and the number of oocytes obtained was the highest (P < 0. 05) . Multiple comparisons showed that Group C4 had lower BMI , follicle-stimulating hormone (FSH) , very low-density lipoprotein ( vLDL) levels , a higher luteinizing hormone and follicle-stimulating hormone ( LH/FSH) ratio compared to Group C1 (P < 0. 05) . Group C4 had lower fasting insulin (FINS) and homeostasis model assessment of insulin resistance ( HOMA-IR) levels compared to Group C3 , and higher high-density lipoprotein-cholesterol (HDL-C) and apolipoprotein A1 (Apo A1) levels compared to Groups C2 and C3 (P < 0. 05) . @*Conclusion@#BMI , the history of previous pregnancy and the number of high-quality embryos were both independent factors for predicting pregnancy outcomes in PCOS patients undergoing FET cycles . Patients with a higher number of high-quality embryos have a higher clinical pregnancy rate during FET cycles .
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Objective @#To develop a prediction model for live birth based on the basic characters of patients after transcervical resection of adhesions (TCRA) undergoing frozen⁃thawed embryo transfer (F⁃ET) .@*Methods @# A total of 491 cycles who went F ⁃ET after TCRA were included. The cycles were randomly divided into training (n = 347) and validation (n = 144) cohorts at 7 ∶ 3 ratio. Relevant factors of live birth were selected via logistic regression analysis based on the data from patients in the training cohort. Multivariate logistic regression analysis was used to establish a nomogram , and then this predict model was calibrated and verified. @*Results @#Age , body mass index (BMI) , miscarriages and endometrial thickness were significantly related to the live birth. The statistical nomogram was built with the area under the ROC cu2rve for the training cohort was 0. 713 (95% CI: 0. 519 - 0. 815) , Hosmer⁃Lemeshow goodness of fit test were χ = 7. 062 , P = 0. 530 , and the slope of calibration curve was close to 1. AUC of validation cohort was 0. 609 (95% CI: 0. 660 - 0. 561) , presenting a well⁃pleasing goodness⁃of⁃fit and stability in this model.@*Conclusion @#Age , BMI , miscarriages and endometrial thickness can better predict the probability of live birth undergoing F ⁃ET after TCRA.
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Objective To investigate pregnancy complications,cesarean delivery and abnormal fetal presentation in women with uterine anomalies.Methods Fifty women diagnosed with uterine malformation and sixty women with normal uterus were retrospectively analyzed .Results The group comprised women with uterine malformation had significantly higher stillbirth or fetal death (P <0.05) and lower rates of the full-term delivery(P <0.05) than that of the control group.The subgroup comprised women with uterine anomalies had significantly higher rates of cesare -an section(P <0.05) and abnormal fetal position(P <0.05) than the control group.The birth weight and length of infant in observation group were significantly lower than those in control group (P <0.05).The rate of placenta abnormalities in uterine anomalies was significantly higher than that in control group (P <0.05).The incidences of premature rupture of membrane,cord entanglement,oligohydramnios,postpartum hemorrhage and neonatal asphyxia in observation group were no significant difference than those in control group .The rate of premature rupture of membrane(PROM) in uterus unicornis was significantly higher than that in other types (P <0.05).Conclusion Compared with women with normal uterus ,the incidences of abnormal fetal position ,the placenta abnormalities,pre-mature delivery,and stillbirth or fetal death in women with congenital uterine anomalies are high .Uterine anomalies are associated with a decrease in the birth weight and length .We should pay more attention to finding the uterine malformation and treatment before pregnancy .Antenatal care should be reinforced in pregnancy women with uterine anomalies.
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Objective To summarize the perioperative nursing of undergoing radical cystectomy with ileal neobladder for the treatment of bladder cancer? Method The nursing measures included preoperative psychological care,bowel and body preparation as well as postoperative nursing of drainage tube,ileal bladder irrigation,bladder functional exercise and basic nursing care? Results No patients suffered from postoperative complications such as infections,urinary fistula and anastomotic leakage? Twenty patients recovered autonomous urination within 3-6 months after the operation? One patient got minor urinal incontinence? Conclusions Cystectomy with orthotopic ileal neobladder trauma contributes to more complications due to large surgical trauma? Therefore,good perioperative care is important for the successful operations?
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Objective To evaluate the association between recurrent miscarriages and insulin resistance.Methods The case-control studies on the association between recurrent spontaneous abortion and insulin resistance from June 1996 to April 2012 were collected from Medline,Elsevier,Chinese Journal Fulltext Database,Chinese Biological Medicine Database,data base of Wanfang,Springer link and EMBASE.RevMan 5.1 software was used for Meta analysis.Results According to the included criteria,7 clinical trials were finally selected.Total 467 cases with recurrent pregnancy loss were enrolled in study group,while 413 women with no history of abnormal pregnancies were enrolled in control group.No significant difference was found in average age and body mass index between the two groups (P ＞ 0.05).Meta analysis results showed that the level of fasting glucose was no statistical difference between study group and control group (WMD =2.27,95％ CI:-1.11 to 5.65,P ＞0.05); fasting insulin level was higher 2.05 mU/L in study group than that of in control group,the difference was statistically significant (WMD =2.05,95％ CI:1.03 to 3.08,P ＜ 0.01).Case number of study group on Homa-insulin resistance ＞ 4.5 was more than that of control group (OR =3.36,95％ CI:1.72 to 6.57,P ＜ 0.01).Case number of study group on glucose/insulin ratio ＜ 4.5 was more than that of the control group,statistical difference was found (OR =3.37,95％ CI:1.90 to 5.99,P ＜ 0.01).Conclusion Insulin resistance is associated with the susceptibility to recurrent miscarriages,and it may contribute to the occurrence of recurrent miscarriages.
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Objective To investigate clinical effect and safety of in vitro maturation(IVM)of human immature oocytes in infertile women with polycystic ovary syndrome by comparing with conventional in vitro fertilization(IVF)and intracytoplasmic sperm injection(ICSI).Methods From Jan.2003 to Dec.2009,157 infertile women with PCOS underwent 162 cycles IVM in Center for Reproductive Medicine,the First Affiliated Hospital of Anhui Medical University.In the mean time,109 patients with PCOS underwent 114 IVF/ICSI cycles as control group 1 and 106 patients with other factors underwent 106 IVF/ICSI cycles as control group 2.Treatment and outcome of pregnancy and infant were compared among those 3 groups.Results No statistically significant difference were found in terms of the positive rate of hCG in urine[35.7％(56/157),42.2％(46/109),44.3％(47/106)],the rate of clinical pregnancy[29.3％ (46/157),37.6％(41/109),41.5％(44/106)],the rate of entopic pregnancy[1.9％(3/157),1.8％ (2/109),0.9％(1/106)],the rate of miscarriage[18.6％(8/43),12.8％(5/39),20.9％(9/43)]and the rate of live-birth[22.3％(35/157),31.2％(34/109),32.1％(34/106)]among three grbups(IVM group,control group 1,control group 2,P ＞ 0.05).The rate of preterm labor,low weight newborn,mean birth weight,ratio of male to female did not show significantly difference among 3 groups(P ＞ 0.05).The average control ovarian stimulation was 6 days,the median dose of gonadotropin(Gn)was 675 IU,and the total hospital cost was(8392 ± 1328)RMB in IVM group,which were statistically lower than those in the other two control groups(P ＜ 0.01).The rate of multiple pregnancy was 4.7％(2/43)and ovarian hyperstimulation syndrome(OHSS)0 in IVM group,which were significantly lower than those in the other control group(P ＜0.01).Conclusion In vitro maturation is an effective treatment in infertile women with PCOS,it could obtain the similar pregnancy outcome and reduce total cost,the dosage of gonadotropinreleasing hormone and rate of OHSS compared with conventional IVF/ICSI.
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Objective To investigate the association between polymorphism of cytochrome P450 subfamily Ⅺ A polypeptide 1 (CYP11A1) gene and polycystic ovarian syndrome (PCOS) in Chinese population. Methods From May 2005 to Dec.2008,290 PCOS cases treated in the First affiliated hospital of Anhui Medical University matched with 344 reproductive women as controls were enrolled in this study. Genotypes of 7 tagging single nucleotide polymorphisms(tSNP,rs12438594,rs4077582,rs9806234,rsl6968477,rs4887139,rs1843090,rsl 1632698)covering CYP11A1 gene (r~2≥0.8) and 3 microsatellite markers (D15S1547,D16S520,D15S1546) were chosed from the phase II database of Han population in HapMap data.Genotype and frequency of allele were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and haplotype of gene polymorphism were analyzed in 290 PCOS cases and 344 controls.Results Among 7 tSNPs and 3 microsatellite markers,the frequency of rs4077582,D15S1547,D15S1546 and rsl 1632698 between two groups reached statistical difference (P =0.010,0.044,0.018 and 0.026).The allele frequency of rs4O77582,rs4887139,rsl843090,D15S1547 and Dl 6S520 showed significant difference between two groups(P=0.002,0.048,0.030,0.001 and 0.024).Among 5 haplotype of CYP11A1(ACGCA13/6/9AG,ACGTA16/6/11AA,GCACG12/8/8AA,GTACA14/4/7GG,GTGCA13/6/7AG),the frequency of GTACA14/4/7GG and ACGCA13/6/9AG were 7.8% (45/580) and 25.3% (147/580) in PCOS group and 11.9% and 19.6% in control group,which showed statistical difference (P< 0.05 ).There was no significant difference in the level of serum androgen at difference genotype from rs4077582 between two groups (P>0.05).Conclusion The polymorphism of CYP11A1 gene was associated with PCOS,however,the relationship between gene sequence covered by tSNP/microsatellite markers and hyperandrogenism of PCOS should be further investigated.
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Objective To investigate the influence on developmental potential of frozen-thawed rabbit oocytes with double assisted activation followed by intracytoplasmic sperm injection (ICSI). Methods A total of rabbit oocytes were collected and thawed after vitrification cryopreservation. Among all oocytes were cultured for 1 hour followed by ICSI. 156 Survived oocytes were divided into 5 groups randomly. I0634 single activation: 30 oocytes were added with calcium ionomycin ( I0634 ) at 5 μmol/L for 5 minutes;SrCl2single activation: 26 oocytes were added with strontium chloride at 10 mmol/L for 10 minutes;10634 double activation: 33 oocytes were activated by I0634 twice;SrCl2 double activation: 28 oocytes were activated by strontium chloride twice. Control group: 39 oocytes were not added with any activators. The rate of fertilization, cleavage and blastocysts formation were observed and compared between various groups. Result The rates of fertilization, cleavage and blastocysts formation were in group of SrCl2 single activation were higher than those of I0634 single activation group without statistical difference (54% vs. 33%, 27% vs. 17%, 8% vs. 3%, P <0.05 ). However, those above rates in double activation by I0634 were higher significantly than those of single I0634 activation (82% vs. 33%, 55% vs. 17%, 15% vs. 3%, P < 0.05). The rates of fertilization (61%) was higher and the rate of cleavage (21%) and blastocysts formation (7%) were lower in group of SrCl2 double activation in comparison with group of SrCl2 single activation without reaching statistical difference (P<0.05 ). Notably, the rates of fertilization, cleavage and blastocysts formation in I0634 double activation group were higher than those in group of SrCl2 double activation with statistical difference (82% vs. 61%, 55% vs. 21%, 15% vs. 7%, P<0.05). Conclusion It might enhance the potential of fertilization of oocytes and early embryo development treated by double activation following ICSI, however, those activated oocytes demonstrate rapid cleavage.
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0bjectlve To examine the metaphase II spindle and chromosome configurations of human oocytes cultured for different times after thawing.MethodsUsing slow.cooling and raid—thawing protocol combined with 0.3 mol/L sucrose and 1.5 mol/L 1,2-propanedio 1(1,2-PROH)to cryoprotect human mature oocytes(n=102),the 64 survival oocytes without abnormal zona pellucida and cytoskeletal were randomly assigned to three groups after thawing:group A:culture 1 hour(n=20),group B:culture 3 hour(n=22),group C:culture 5 hours(n=22),the flesh oocytes served as control group(n=18).Immunocytochefifical staining and fluorescence microscopy were used to assess the morphology of the metaphase II spindle and chromosome.Results(1)The normal spindle rates of groups A,B and C were 10％(2/20),46％(10/22)and 41％(9/22)respectively,significantly decreased compared with control group(83％,15/18;P<0.05).The rates of absent spindle in group A(45％,9/20)was significantly higher than control group(6％,1/18;P<0.01).Also,the rates of absent spindle in group A was higher than groups B(14％,3/20)and C(14％,3/20;P<0.05).However,no significant differences were observed in groups B and C(P>0.05).(2)A significant increase in abnormal chromosome rate was observed in group A(30％,6/20)compared to groups B(68％,15/22),C(64％,14/22)and control group(78％,14/18;尸<0.05).No differences in chromosome morphology were observed in groups B,C and control group(P>0.05).Conclusions The cryoproteetant protocol leads to a deleterious effect on the organization of the meiotic spindle and chromosome at MI stage.The 3—5 hours post—thawing incubation could permit restoration of the meiotic spindles and chromosome.
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Objective To understand the expressions of transforming growth factor ?1(TGF-?1) and its receptors(T?R) in villi and decidua during early pregnancy and their effects on early embryo development, and explore the mechanism of spontaneous abortion. Methods By immunohistochemical technique, expressions of TGF-?1 and its receptors were determined in villi and decidua from 10 cases of spontaneous abortion women after in vitro fertilization and embryo transfer (IVF-ET group), 20 cases of spontaneous abortion women (spontaneous abortion group), and 20 cases of normal early pregnancy women (control group). Results TGF-?1 and its receptors had high expressions in decidua cells, villus glands and interstitial cytoplasm in all three groups. The average light density of the expressions of TGF-?1 in the villi and decidua of IVF-ET, spontaneous abortion and control groups were 0.167 and 0.199, 0.198 and 0.201, 0.277 and 0.274, respectively. The intensity of T?R-Ⅰ in the villi and decidua of IVF-ET, spontaneous abortion and control groups were 0.144 and 0.150, 0.202 and 0.201, 0.238 and 0.281, respectively. And the expressions of T?R-Ⅱ in the villi and decidua of IVF-ET, spontaneous abortion and control groups were 0.199 and 0.145, 0.153 and 0.156, 0.300 and 0.301, respectively. The differences between the control group and both abortion groups were all significant (P
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Objective To evaluate diagnosis of cord entanglem ent and fetus safety by B ultra-sonic and nemodromometer. Methods 86 pregnant women were diagnosed as cord entanglement in late pregnancy by B ultrasonic examination and Doppler blood flow analysis of the S/D ratio from ma r to May, 1999. The pregnancy outcomes were followed up. Cord entanglement,es -pecially the high blood circulating resistance, suggested that there was h igh risk of fetal distress. The obstetrician would manage the delivery stage pro perly according to the results of Doppler examination.Results 7 2 women were proved cord entanglement after delivery wity coincident rate of 83. 7%. Five women were delivered by cesarean section and forceps among 11 cases of abno rmal S/D ratio, and the rate of operative delivery in group with abnormalS/D rat io was significantly higher than that of the group with normal S/D ratio (P ＜0.05). Conclusions The combined examination can improve the rate of diagnosis of cord entanglement and abnormal S/D ratio, forecast the feta l distress, reduce birth rate of infant with lower Apgar score and reduce the ces arean section rate due to social factor.