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1.
Clin Lab ; 67(11)2021 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-34758230

RESUMEN

BACKGROUND: Uterine corpus endometrial carcinoma (UCEC) is the third most prevalent female reproductive system malignant tumor with poor prognosis, particularly at advanced stage. On the other hand, recent studies have reported the prognostic role of long non-coding RNAs (lncRNAs) in UCEC. The aim of this study was to determine the immune-related lncRNA signature for predicting overall survival (OS) in UCEC patients. METHODS: The genomic data and clinical information of UCEC patients were extracted from the Cancer Genome Atlas. Pearson's correlation analysis was carried out to identify the immune-related lncRNAs. Univariate and multivariate Cox regression analyses were conducted to obtain the prognostic lncRNAs from the immune-related lncRNAs for the construction of the prognostic signature. Afterwards, the UCEC patients were divided into high-risk and low-risk groups. The prognostic value of the signature was assessed by survival, receiver operating characteristic (ROC), and nomogram analyses. Finally, the immune status for high-risk and low-risk groups was evaluated by the ESTIMATE algorithm. RESULTS: A total of 13 immune-related lncRNAs (AC108860.2, AC015849.5, AL592494.3, LINC01234, U91319.1, AC092969.1, AL356133.2, AC103563.2, AL138962.1, AC138965.1, LINC01687, AC091987.1, and MIR7-3HG) were finally identified for the construction of the prognostic signature. Patients in the high-risk group had worse prognosis than those in the low-risk group. The prognostic signature was confirmed as an independent prognostic factor through the multivariate Cox regression analysis. The nomogram based on the prognostic signature and clinicopathologic features was constructed with a superior overall predictive power to evaluate the survival outcomes in UCEC patients. Finally, according to the ESTIMATE algorithm results, we discovered different immune statuses in the low-risk and high-risk groups. CONCLUSIONS: The immune-related lncRNA signature for the assessment of the OS of UCEC patients had a good practical value.


Asunto(s)
Neoplasias Endometriales , ARN Largo no Codificante , Biomarcadores de Tumor/genética , Neoplasias Endometriales/genética , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Pronóstico , ARN Largo no Codificante/genética
2.
Clin Proteomics ; 15: 19, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29719495

RESUMEN

BACKGROUND: Semen cryopreservation has been widely applied in assisted reproductive technologies and sperm bank, but it causes considerable impairments on sperm quality. It is necessary to find an evaluation indicator for determining the sperm-freezing tolerance. METHODS: The glycocalyx of good freezability ejaculates was compared with poor freezability ejaculates by lectin microarray. The significant different lectins were validated by flow cytometry (FACS). To analyze the relationship between the potential biomarker and the tolerance of sperm to cryopreservation, 60 samples with different recovery rates were collected and detected the lectin-binding intensity by FACS. The receiver operating characteristic (ROC) curve was analyzed to test the capability of the lectin as a potential biomarker for detecting the sperm freezablility. RESULTS: ABA and DSL were found to develop significant differences between them. Further validation showed that ABA was significantly negative correlated with the sperm recovery rates (r = - 0.618, P < 0.000) and could be a potential biomarker for predicting sperm freezability (AUC = 0.733 ± 0.067, 95% CI 0.601 - 0.865, P < 0.01). CONCLUSION: ABA could be a potential biomarker for predicting sperm freezability. It will help to reduce sperm-freezing recovery tests and improve the efficiency of cryopreservation in human sperm bank.

3.
Reprod Biomed Online ; 37(1): 25-32, 2018 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-29703434

RESUMEN

RESEARCH QUESTION: Are miRNAs found in follicular fluid related to blastocyst formation from the corresponding oocytes? DESIGN: In this study, 91 individual follicular fluid samples from single follicles containing mature oocytes from 91 women were collected and classified into group 1 (n = 38) with viable blastocysts, and group 2 (n = 53) with no blastocyst. TaqMan human miRNA cards and quantitative reverse transcription polymerase chain reaction were used to identify differently expressed follicular fluid miRNAs between the two groups. RESULTS: We found MIR-663B to be significantly differentially expressed in follicular fluid of oocytes that yielded viable blastocysts versus those that did not develop into blastocysts (14.16 ± 7.00 versus 23.68 ± 17.02; P = 0.019), as well as for those which develop into blastocysts with good morphology versus those with poor morphology (11.69 ± 3.49 versus 20.16 ± 9.33; P = 0.003). CONCLUSIONS: MIR-663B expression levels in human follicular fluid samples were significantly negatively related to viable blastocyst formation and may become an objective evaluation criterion for embryo development potential after IVF.


Asunto(s)
Blastocisto/metabolismo , Desarrollo Embrionario/genética , Líquido Folicular/metabolismo , MicroARNs/metabolismo , Adulto , Femenino , Humanos , MicroARNs/genética , Oocitos/metabolismo , Oogénesis/fisiología , Inyecciones de Esperma Intracitoplasmáticas
4.
Cell Physiol Biochem ; 44(3): 1093-1105, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-29179212

RESUMEN

BACKGROUND/AIMS: LukS-PV is a component of Panton-Valentine leukocidin (PVL). We have previously demonstrated that LukS-PV potently promoted differentiation and induced apoptosis in THP-1 cells. However, the precise mechanisms of these actions remain unknown. MicroRNAs (miRs) play important roles in cellular differentiation and apoptosis. This study aimed to investigate the role of miR-125a-3p in LukS-PV-regulated differentiation and apoptosis and its underlying mechanism in THP-1 cells. METHODS: MicroRNA profiling analyses were conducted to determine differential miRNA expression levels in THP-1 cells treated with LukS-PV. Cell differentiation and apoptosis were measured in THP-1 cells by gain-of-function and loss-of-function experiments. Bioinformatics analysis and luciferase reporter assays were used to confirm the targets of miR-125a-3p. The effects of the miR-125a-3p targets on cellular differentiation were determined by knocking them down. RESULTS: MiR-125a-3p was up-regulated after treating the human monocytic leukaemia cell line THP-1 with LukS-PV. In addition, miR-125a-3p positively regulated apoptosis and differentiation in THP-1 cells treated with LukS-PV. Concordantly, luciferase reporter assays confirmed that neurofibromatosis type 1 (NF1) and B-cell lymphoma 2 (Bcl-2) were direct target genes of miR-125a-3p. Moreover, NF1 knockdown in THP-1 cells significantly promoted differentiation in vitro. Finally, the extracellular signal-regulated kinase (ERK) pathway, a downstream target of NF1, was activated after NF1 knockdown. CONCLUSIONS: These findings confirm that miR-125a-3p is involved in LukS-PV-mediated cell differentiation and apoptosis in THP-1 cells.


Asunto(s)
Apoptosis/efectos de los fármacos , Toxinas Bacterianas/farmacología , Diferenciación Celular/efectos de los fármacos , Exotoxinas/farmacología , Leucocidinas/farmacología , MicroARNs/metabolismo , Neurofibromina 1/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Regiones no Traducidas 3' , Antagomirs/metabolismo , Toxinas Bacterianas/genética , Toxinas Bacterianas/metabolismo , Secuencia de Bases , Caspasa 3/metabolismo , Línea Celular , Regulación hacia Abajo/efectos de los fármacos , Exotoxinas/genética , Exotoxinas/metabolismo , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Puntos de Control de la Fase G1 del Ciclo Celular/efectos de los fármacos , Humanos , Leucocidinas/genética , Leucocidinas/metabolismo , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , MicroARNs/antagonistas & inhibidores , MicroARNs/genética , Neurofibromina 1/antagonistas & inhibidores , Neurofibromina 1/genética , Fosforilación/efectos de los fármacos , Proteínas Proto-Oncogénicas c-bcl-2/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/farmacología , Alineación de Secuencia , Transducción de Señal/efectos de los fármacos , Transcriptoma/efectos de los fármacos , Proteína X Asociada a bcl-2/metabolismo
5.
Clin Invest Med ; 40(3): E135-E145, 2017 06 26.
Artículo en Inglés | MEDLINE | ID: mdl-28653615

RESUMEN

PURPOSE: The purpose of this study was to perform a meta-analysis comparing the rates of uterine rupture, and other maternal and neonatal complications, between women who undergo a trial of labour (TOL) after a prior Cesarean delivery and those to undergo elective repeat Cesarean delivery (ERCD). SOURCE: Medline, Cochrane, EMBASE and Google Scholar were searched until May 6, 2015 using the keywords/phrases: trial of labour, Cesarean section, elective, repeat, pregnancy and vaginal birth. Randomized controlled trials (RCTs), two-arm prospective studies, one-arm studies and retrospective studies were included. The primary outcome was uterine rupture. PRINCIPAL FINDINGS: Sixteen studies were included in the meta-analysis. TOL after prior Cesarean delivery was associated with higher odds of uterine rupture as compared with ERCD (Peto odds ratio [OR] = 4.685, 95% confidence interval [CI]: 3.077 to 7.133, p < 0.001). TOL was associated with a higher rate of endometritis, a lower rate of hysterectomy, and a lower rate of respiratory problems in newborns. There were no differences between the groups with respect to neonatal intensive care unit admissions, postpartum hemorrhage, thromboembolic disease, sepsis and neonatal mortality. CONCLUSIONS: TOL may be associated with a higher risk of uterine rupture and endometritis, but lower risk of hysterectomy and neonatal respiratory problems than ERCD.


Asunto(s)
Cesárea/efectos adversos , Mortalidad Infantil , Endometritis/epidemiología , Endometritis/etiología , Femenino , Humanos , Lactante , Recién Nacido , Embarazo , Resultado del Embarazo , Esfuerzo de Parto , Rotura Uterina/epidemiología , Rotura Uterina/etiología
6.
Reprod Biol Endocrinol ; 13: 114, 2015 Oct 06.
Artículo en Inglés | MEDLINE | ID: mdl-26444856

RESUMEN

BACKGROUND: The purpose of this study was to test the hypothesis that estradiol (E2) level on day 3 may be associated with in vitro fertilization (IVF) outcomes. METHODS: The records of patients who received clomiphene citrate 100 mg/day plus human menopausal gonadotropin 150 IU/day from day 3 and received frozen-thawed embryo transfers were reviewed. Patients were divided into three groups: group A (E2 ≤30 pg/ml), group B (30< E2 ≤50 pg/ml), and group C (E2 >50 pg/ml). A total of 1080 cycles from 941 patients were included. RESULTS: The number of eggs and MII oocytes were less in group C than group A (both, P = 0.001). The embryo implantation (P = 0.006) and clinical pregnancy rates (P = 0.036) were lower in group C than group B, and the rates were similar between group A and B. CONCLUSION: Maintaining the serum E2 level from 30 to 50 pg/ml may result in a higher clinical pregnancy rate in IVF cycles.


Asunto(s)
Clomifeno/farmacología , Transferencia de Embrión/métodos , Estradiol/sangre , Fertilización In Vitro/métodos , Inducción de la Ovulación/métodos , Índice de Embarazo , Adulto , Biomarcadores/sangre , Femenino , Fertilización In Vitro/efectos de los fármacos , Humanos , Embarazo , Índice de Embarazo/tendencias , Estudios Retrospectivos , Resultado del Tratamiento
7.
Am J Obstet Gynecol ; 212(1): 53.e1-10, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25046807

RESUMEN

OBJECTIVE: To determine whether previous salpingectomy is associated with serum antiMüllerian hormone (AMH) level and ovarian reserve in women under 40 years presenting for in vitro fertilization and embryo transfer. STUDY DESIGN: We retrospectively compared serum AMH levels measured on the ovulation induction initiation day in patients with unilateral salpingectomy, bilateral salpingectomy, and no tubal surgery, and examined the relationship with length of time after surgery and in vitro fertilization and embryo transfer parameters. RESULTS: A total of 198 women were included; 83 received unilateral salpingectomy, 41 bilateral salpingectomy, and 74 no tubal surgery. The baseline characteristics of the groups were similar. The mean AMH level was significantly higher in women without tubal surgery as compared with those with bilateral salpingectomy (183.48 vs 127.11 fmol/mL; P ≤ .037). The mean follicle stimulation hormone level was significantly lower in women without surgery as compared with those with bilateral salpingectomy (7.85 vs 9.13 mIU/mL; P = .048). No significant differences in duration of gonadotropin therapy, amount of gonadotropin used, estradiol level on the human chorionic gonadotropin injection day, thickness of the endometrium, number of oocytes retrieved, number of 2-pronuclei, viable embryos, and good quality embryos were found between the 3 groups. AMH level was not correlated with the number of oocytes or age in women that had undergone unilateral or bilateral salpingectomy. CONCLUSION: Salpingectomy is associated with decreased AMH level and increased follicle stimulation hormone in women seeking in vitro fertilization, though AMH level is not correlated with the number of oocytes retrieved in patients that have undergone unilateral or bilateral salpingectomy. These results suggest that salpingectomy is associated with decreased ovarian reserve.


Asunto(s)
Hormona Antimülleriana/sangre , Reserva Ovárica , Salpingectomía , Adulto , Transferencia de Embrión , Femenino , Fertilización In Vitro , Humanos , Estudios Retrospectivos
8.
J Assist Reprod Genet ; 32(11): 1597-605, 2015 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-26463877

RESUMEN

PURPOSE: Assessment of embryo viability is a crucial component of in vitro fertilization and currently relies largely on embryo morphology and cleavage rate. Because morphological assessment remains highly subjective, it can be unreliable in predicting embryo viability. This study investigated the metabolomic profiling of embryo culture media using near-infrared (NIR) spectroscopy for predicting the implantation potential of human embryos in frozen-thawed embryo transfer (FET) cycles. METHODS: Spent embryo culture media was collected on day 4 after thawed embryo transfer (n = 621) and analysed using NIR spectroscopy. Viability scores were calculated using a predictive multivariate algorithm of fresh embryos with known pregnancy outcomes. RESULTS: The mean viability indices of embryos resulting in clinical pregnancy following FET were significantly higher than those of non-implanted embryos and differed between the 0, 50, and 100 % implantation groups. Notably, the 0 % group index was significantly lower than the 100 % implantation group index (-0.787 ± 0.382 vs. 1.064 ± 0.331, P < 0.01). To predict implantation outcomes, we examined the area under the ROC curve (AUCROC), which was significantly higher for the viability than for the morphology score (0.94 vs. 0.55; P < 0.01); however, the AUCROCs for the composite and viability scores did not differ significantly (0.92 vs. 0.94; P > 0.05). CONCLUSIONS: NIR metabolomic profiling of thawed embryo culture media is independent of morphology and correlates with embryo implantation potential in FET cycles. The viability score alone or in conjunction with morphologic grading is a more objective marker for implantation outcome in FET cycles than morphology alone.


Asunto(s)
Criopreservación/métodos , Medios de Cultivo/análisis , Implantación del Embrión , Transferencia de Embrión/métodos , Metabolómica/métodos , Adulto , Medios de Cultivo/química , Técnicas de Cultivo de Embriones , Femenino , Fertilización In Vitro/métodos , Humanos , Recién Nacido , Masculino , Embarazo , Resultado del Embarazo , Curva ROC , Espectroscopía Infrarroja Corta
9.
Biomed Environ Sci ; 28(10): 769-72, 2015 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-26582099

RESUMEN

The study objectives were to investigate the relationship between early exposure to genistein and obesity in young adulthood and to evaluate changes in reproductive health during puberty and adulthood following in utero exposure to genistein. Thirty-two female rats were randomized into four groups; low dose 400 mg genistein/kg diet group (LG), mid-dose 1200 mg genistein/kg diet group (MG), high dose 3600 mg genistein/kg diet group (HG), and control group without genistein diet (CON). Rats were fed genistein at the beginning of pregnancy along with a high-fat diet. Pups were sacrificed at week 4 and week 8 after birth. High performance liquid chromatography (HPLC) results showed a correlation between maternal genistein intake and genistein concentration in pups' plasma. Compared to CON, body weight reduced significantly in male HG group at week 8. No statistical differences were found in plasma estradiol (E2), testosterone (T), interleukin (IL)-6, and C-reactive protein (CRP) levels with early genistein exposure. Furthermore, uterine histopathology showed notable changes in groups HG and MG compared with CON at week 4 and week 8. In conclusion, maternal genistein supplement could reduce body weight in male pups and alter uterine histopathology in female pups.


Asunto(s)
Peso Corporal/efectos de los fármacos , Genisteína/farmacología , Efectos Tardíos de la Exposición Prenatal , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Grasas de la Dieta/administración & dosificación , Femenino , Genisteína/administración & dosificación , Genisteína/sangre , Masculino , Fenómenos Fisiologicos Nutricionales Maternos , Embarazo , Distribución Aleatoria , Ratas , Útero/crecimiento & desarrollo
10.
Sheng Li Xue Bao ; 66(6): 709-17, 2014 Dec 25.
Artículo en Zh | MEDLINE | ID: mdl-25516520

RESUMEN

Cardiac extracellular matrix (ECM), generated from the process of decellularization, has been widely considered as an ideal source of biological scaffolds. However, current ECM preparations are generally difficult to be applied to generate cardiac tissue. Our research was aimed to improve decellularization protocols to prepare cardiac ECM slices. Adult murine ventricular tissues were embedded in low melting agarose and cut into 300 µm slices, and then were divided randomly into three groups: normal cardiac tissue, SDS treated group (0.1% SDS) and SDS+Triton X-100 treated group (0.1% SDS+0.5% Triton X-100). Total RNA content and protein content quantification, HE staining and immunostaining were used to evaluate the removal of cell components and preservation of vital ECM components. Furthermore, murine embryonic stem cell-derived cardiomyocytes (mES-CMs) and mouse embryonic fibroblasts (MEFs) were co-cultured with ECM slices to evaluate biocompatibility. The relative residual RNA and protein contents of ECM slices significantly decreased after decellularization. HE staining showed that SDS+Triton X-100 treatment better destroyed cellular structure and removed nuclei of ECM slices, compared with SDS treatment. Immunostaining showed that collagen IV and laminin were better preserved and presented better similarity to original cardiac tissue in ECM slices acquired by SDS+Triton X-100 treatment. However, collagen IV and laminin were significantly decreased and arranged disorderly in SDS treated group. We observed effective survival (≥ 12 days) of MEFs and mES-CMs on ECM slices acquired by SDS+Triton X-100 treatment, and signs of integration, whereas those signs were not found in SDS treated group. We concluded that, compared with traditional SDS method, new combined protocol (SDS+Triton X-100) generated ECM slices with better component and structural preservation, as well as better biocompatibility.


Asunto(s)
Matriz Extracelular/química , Ventrículos Cardíacos/citología , Ingeniería de Tejidos/métodos , Andamios del Tejido , Animales , Ratones , Octoxinol , Dodecil Sulfato de Sodio
11.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 30(5): 522-7, 2013 Oct.
Artículo en Zh | MEDLINE | ID: mdl-24078563

RESUMEN

OBJECTIVE: To investigate the efficiency of multi-round fluorescence in situ hybridization (FISH) and its influencing factors in preimplantation genetic diagnosis (PGD). METHODS: A total of 48 couples accepted PGD because of various reasons: 24 with Robertsonian translocations, 16 with reciprocal translocations, 2 with pericentric inversions, one with advanced maternal age who had a previous liveborn of Down syndrome, 3 suffered from sex chromosome abnormalities and 2 repeated spontaneous miscarriages. After 72 retrieval cycles, 432 cleavage stage embryos with more than six cells were biopsied on day three. Only intact nuclei (396) were hybridized in order to verify the chromosomal status of the individual embryos. If previous FISH has failed to give conclusive results while the nuclei remained undamaged, the nuclei were hybridized once again. A total of 870 times of hybridization were conducted to 396 nuclei. Signal identification rates of each round as well as the influence of different probes to the hybridization efficiency were compared. Factors leading to inconclusive FISH results were analyzed as well. RESULTS: Five hundred and thirty five out of 870 hybridizations gave identifiable signals (61.5%). The second and third round FISH showed the best signals with an identification rate of 71.8% and 77.4%, respectively, which were significantly higher than those of the first round (52.8%, P < 0.01), the fourth round (55.8%, P < 0.05, P < 0.01), the fifth round (54.5%, P < 0.05) and the sixth round (27.3%, P < 0.01). The identification rate of centromere specific probe signals (CEP group) was 80.3% and the former three rounds in this group got the best quality of signals with an identification rate of 85.7%, 85.1% and 88.0%, respectively, which was significantly higher than that of the latter three rounds. The identification rate of other probe was much lower than with the CEP probe (55.2% vs. 80.3%, P < 0.01) and the best quality of signal in this group was achieved in the fifth round (72.7%), followed by the second round (66.1%) and the third round (63.8%). The identification rate of the first round (50.3%) and the sixth round (22.2%) were significantly lower compared with the second round (P < 0.01). During the 6 rounds of FISH, 335 hybridizations did not give conclusion results (38.5%, 335/870). The main cause of unidentification was weak signals (20.9%, 182/870). Other common factors included background interference (7.6%, 66/870) and failed hybridization (6.1%, 53/870). Rare causes included nucleus damage (1.8%, 16/870), nucleus loss (1.1%, 10/870) and signal split/overlap (0.9%, 8/870). CONCLUSION: Multi-round FISH can improve the utility of single nucleus in PGD and the former three rounds have the highest efficiency. The hybridization effect of CEP is better than other probe. Poor signal quality is the common cause of unidentification results.


Asunto(s)
Pruebas Genéticas/métodos , Hibridación Fluorescente in Situ/métodos , Diagnóstico Preimplantación/métodos , Femenino , Humanos , Masculino , Embarazo , Diagnóstico Prenatal , Translocación Genética
12.
Front Endocrinol (Lausanne) ; 14: 1292723, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38352249

RESUMEN

Background: The molecular mechanisms underlying window of implantation (WOI) displacement in patients with recurrent implantation failure (RIF) remain unclear. This study aims to explore the transcriptomic signatures of endometrium with normal and displaced WOIs and to identify the causes of endometrial receptivity (ER) abnormalities and WOI displacement in RIF patients. Methods: In this study, 40 RIF patients were recruited and underwent personalized embryo transfer (pET) guided by the predicted results of endometrial receptivity diagnosis (ERD) model. Transcriptome analysis of endometrium from patients with clinical pregnancies after pET was performed to identify differentially expressed genes (DEGs) associated with WOI displacement. Gene expression data from HRT and natural cycle endometrium were compared to identify specific gene expression patterns of ER-related genes during WOI. Results: The ERD results indicated that 67.5% of RIF patients (27/40) were non-receptive in the conventional WOI (P+5) of the HRT cycle. The clinical pregnancy rate in RIF patients improved to 65% (26/40) after ERD-guided pET, indicating the effectiveness of transcriptome-based WOI prediction. Among the 26 patients with clinical pregnancy, the gene expression profiles of P+5 endometrium from advanced (n=6), normal (n=10) and delayed (n=10) WOI groups were significantly different from each other. Furthermore, 10 DEGs identified among P+5 endometrium of 3 groups were involved in immunomodulation, transmembrane transport and tissue regeneration, which could accurately classify the endometrium with different WOIs. Additionally, a large number of ER-related genes showed significant correlation and similar gene expression patterns in P+3, P+5, and P+7 endometrium from HRT cycles and LH+5, LH+7, and LH+9 endometrium from natural cycles. Conclusion: Our study shows that ER-related genes share similar gene expression patterns during WOI in both natural and HRT cycles, and their aberrant expression is associated with WOI displacements. The improvement of pregnancy outcomes in RIF patients by adjusting ET timing according to ERD results demonstrates the importance of transcriptome-based endometrial receptivity assessment and the clinical efficiency of ERD model.


Asunto(s)
Implantación del Embrión , Endometrio , Embarazo , Femenino , Humanos , Endometrio/metabolismo , Implantación del Embrión/genética , Perfilación de la Expresión Génica , Transcriptoma , Resultado del Embarazo
13.
Clin Dev Immunol ; 2012: 831010, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22162720

RESUMEN

The human zona pellucida glycoprotein-3 (hZP3) by virtue of its critical role during fertilization has been proposed as a promising candidate antigen to develop a contraceptive vaccine. In this direction, it is imperative to map minimal motifs of the B cell epitopes (BCEs) so as to avoid ZP-specific oophoritogenic T cell epitopes (TCEs) in the ZP3-based immunogens. In this study, based on known results of mapping marmoset and bonnet monkey ZP3 (mstZP3 and bmZP3), two predictable epitopes(23-30 and 301-320) on hZP3 were first confirmed and five minimal motifs within four epitopes on hZP3 were defined using serum to recombinant hZP3a(22-176) or hZP3b(177-348) as well as a biosynthetic peptide strategy. These defined minimal motifs were QPLWLL(23-28) for hZP3(23-30), MQVTDD(103-108) for hZP3(93-110), EENW(178-181) for hZP3(172-190), as well as SNSWF(306-310) and EGP(313-315) for hZP3(301-320), respectively. Furthermore, the antigenicity of two peptides for hZP3(172-187) and hZP3(301-315) and specificity of the antibody response to these peptides were also evaluated, which produced high-titer antibodies in immunized animals that were capable of reacting to ZP on human oocytes, r-hZP3b(177-348) protein, as well as r-hZP3(172-190), r-hZP3(303-310), and r-hZP3(313-320) epitope peptides fused with truncated GST188 protein.


Asunto(s)
Proteínas del Huevo/inmunología , Epítopos de Linfocito B/inmunología , Glicoproteínas de Membrana/inmunología , Receptores de Superficie Celular/inmunología , Zona Pelúcida/metabolismo , Animales , Proteínas del Huevo/metabolismo , Mapeo Epitopo , Humanos , Masculino , Glicoproteínas de Membrana/metabolismo , Conejos , Receptores de Superficie Celular/metabolismo , Zona Pelúcida/inmunología , Glicoproteínas de la Zona Pelúcida
14.
Asian J Androl ; 24(1): 62-66, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-34135171

RESUMEN

For infertility treatment, the selection of in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) is decided by multiplying indicators (including fallopian tube factors, semen count, and semen motility), except for sperm morphology. In this study, we conducted a retrospective analysis, from implantation to birth, over a period of 5 years. A total of 1873 couples with primary or secondary fallopian tube factors and an increased defective sperm morphology rate (DSMR) were divided into different groups to receive IVF or ICSI cycles. By comparing the outcomes, we found that the F1 group (DSMR <96%, IVF group 1) had higher cleavage rate, biochemical pregnancy rate, clinical pregnancy rate, and live birth rate than the F3 group (DSMR >98%, IVF group 3; P < 0.05). In contrast, there was no significant difference in the ICSI subgroups. Furthermore, a comparison of the outcomes between IVF and ICSI showed that the S3 group (DSMR >98%, ICSI group 3) had higher cleavage rate (P < 0.001), biochemical pregnancy rate (P < 0.05), clinical pregnancy rate (P < 0.05) and live birth rate (P < 0.05) than the F3 group. However, the ICSI subgroup had a lower two pronuclei fertilization rate than the IVF subgroup (P < 0.05). Our data suggest that the sperm morphology should also be considered when selecting IVF or ICSI combined with other semen parameters before the first assisted reproductive technologies (ART) cycle, especially for males with severe sperm defects.


Asunto(s)
Fertilización In Vitro , Inyecciones de Esperma Intracitoplasmáticas , Femenino , Humanos , Masculino , Embarazo , Índice de Embarazo , Técnicas Reproductivas Asistidas , Estudios Retrospectivos , Espermatozoides
15.
Contemp Clin Trials Commun ; 28: 100928, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-35669489

RESUMEN

Background: Today, approximately 10% of participants in assisted reproductive technology (ART) are defined as having recurrent implantation failure (RIF). Recent studies show that endometrial receptivity array can improve pregnancy and implantation rates by nearly 20% in women with RIF. However, these studies are limited, with little published data in the Chinese population. Recently, we have established a transcriptome-based endometrial receptivity assessment (Tb-ERA) method of predicting the endometrial window of implantation (WOI) using transcriptome-profiling data of different phases of the menstrual cycle from healthy fertile Chinese women by RNA-Seq. It is meaningful to conduct a randomized controlled trial (RCT) to assess the clinical efficiency of Tb-ERA in Chinese patients with RIF. Methods: In this RCT, a total of 200 RIF patients will be recruited and randomized into 2 groups. Patients in the Tb-ERA group will undergo a Tb-ERA test, after which embryo transfer time will be adjusted according to Tb-ERA results and embryo transfer will be performed again in the next cycle. Patients in the control group will not receive any interventions until the next transfer cycle. We will perform statistical analysis on both groups at the primary endpoint (clinical-pregnancy rate) and at secondary endpoints (rate of WOI displacement, embryo implantation, biochemical pregnancy, early abortion, and ectopic pregnancy). Implications: This study aims to evaluate the effectiveness of our Tb-ERA test in Chinese RIF patients and to determine that whether Tb-ERA could improve the clinical-pregnancy rate in these RIF patients. Trial registration: NCT04497558, registered August 4, 2020.

16.
Asian J Androl ; 24(3): 260-265, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35532568

RESUMEN

Obtaining high-quality embryos is one of the key factors to improve the clinical pregnancy rate of assisted reproductive technologies (ART). So far, the clinical evaluation of embryo quality depends on embryo morphology. However, the clinical pregnancy rate is still low. Therefore, new indicators are needed to further improve the evaluation of embryo quality. Several studies have shown that the decrease of sperm-specific protein actin-like 7A (ACTL7A) leaded to low fertilization rate, poor embryo development, and even infertility. The aim of this study was to study whether the different expression levels of ACTL7A on sperm can be used as a biomarker for predicting embryo quality. In this study, excluding the factors of severe female infertility, a total of 281 sperm samples were collected to compare the ACTL7A expression levels of sperms with high and low effective embryo rates and analyze the correlation between protein levels and in-vitro fertilization (IVF) laboratory outcomes. Our results indicated that the ACTL7A levels were significantly reduced in sperm samples presenting poor embryo quality. Furthermore, the protein levels showed a significant correlation with fertilization outcomes of ART. ACTL7A has the potential to be a biomarker for predicting success rate of fertilization and effective embryo and the possibility of embryo arrest. In conclusion, sperm-specific protein ACTL7A has a strong correlation with IVF laboratory outcomes and plays important roles in fertilization and embryo development.


Asunto(s)
Fertilización In Vitro , Técnicas Reproductivas Asistidas , Biomarcadores/metabolismo , Femenino , Fertilización , Humanos , Masculino , Embarazo , Índice de Embarazo , Espermatozoides/metabolismo
17.
BMJ Open ; 12(7): e063030, 2022 07 13.
Artículo en Inglés | MEDLINE | ID: mdl-35831058

RESUMEN

INTRODUCTION: Conventional intracytoplasmic sperm injection (ICSI) is a widely used treatment for couples with severe male infertility. However, there are controversies regarding the selection and the damage to gametes during the ICSI procedure. Although preimplantation genetic testing for aneuploidies (PGT-A) can give genetic information about embryos for transfer and improve fertility rate, and it is widely used in women with recurrent spontaneous abortion or advanced age, PGT-A is not only more expensive but also has unclear effectiveness with respect to the improvement of fertility rate among couples with severe male infertility. High-quality, well-powered randomised clinical trials (RCTs) comparing ICSI+PGT-A and ICSI are lacking. METHODS AND ANALYSIS: This is a protocol for a multicenter, open-label RCT in four reproductive medical centers qualified for PGT technique in China. We will study couples with severe male infertility scheduled for their fertility treatment. After the blastocyst culture, eligible participants are randomised to the ICSI+PGT-A group or the conventional ICSI group in a 1:1 ratio. Other assisted reproductive procedures are similar and parallel between the two groups. The primary outcome will be live birth rate and cumulative live-birth rate . Secondary outcomes will be embryo implantation rate, biochemical pregnancy rate, clinical pregnancy rate, spontaneous abortion rate, ongoing pregnancy rate, preterm birth rate, fetal chromosomal abnormality rate, birth defect rate and treatment complications. To demonstrate or refute a difference between the two groups, we plan to include 188 participants in each group; taking consideration of 20% of dropout, the total target sample size is 450. ETHICS AND DISSEMINATION: Ethical approval was obtained from International Peace Maternity and Child Health Hospital of Shanghai Jiao Tong University Medical Science Research Ethics Committee (GKLW2016-16). Informed consent will be obtained from each participant. The findings will be disseminated to the public through conference presentations and publication in peer-reviewed scientific journals. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov, NCT02941965.


Asunto(s)
Aborto Espontáneo , Infertilidad Masculina , Aborto Espontáneo/genética , Aneuploidia , Niño , China , Femenino , Fertilización In Vitro , Pruebas Genéticas/métodos , Humanos , Recién Nacido , Infertilidad Masculina/genética , Infertilidad Masculina/terapia , Nacimiento Vivo , Masculino , Estudios Multicéntricos como Asunto , Embarazo , Índice de Embarazo , Ensayos Clínicos Controlados Aleatorios como Asunto
18.
Cell Death Discov ; 8(1): 495, 2022 Dec 22.
Artículo en Inglés | MEDLINE | ID: mdl-36550096

RESUMEN

KDM5C is a histone H3K4-specific demethylase, which has been shown to play a key role in biological disease and development. However, the role of KDM5C in trophoblasts at early pregnancy is currently unknown. Here, we showed that KDM5C was upregulated in placental trophoblasts from recurrent miscarriage (RM) patients compared with healthy controls (HCs). Trophoblast proliferation and invasion was inhibited by KDM5C overexpression and was promoted by KDM5C knockdown. Transcriptome sequencing revealed that elevated KDM5C exerted anti-proliferation and anti-invasion effects by repressing the expression of essential regulatory genes. The combination analysis of RNA-seq, ChIP-seq and CUT&Tag assay showed that KDM5C overexpression leads to the reduction of H3K4me3 on the promoters and the corresponding downregulation of expression of several regulatory genes in trophoblasts. Among these genes, TGFß2 and RAGE are essential for the proliferation and invasion of trophoblasts. Importantly, overexpression of KDM5C by a systemically delivered KDM5C adenovirus vector (Ad-KDM5C) promoted embryo resorption rate in mouse. Our results support that KDM5C is an important regulator of the trophoblast function during early pregnancy, and suggesting that KDM5C activity could be responsible for epigenetic alterations seen RM disease.

19.
Zhonghua Fu Chan Ke Za Zhi ; 46(5): 355-9, 2011 May.
Artículo en Zh | MEDLINE | ID: mdl-21733372

RESUMEN

OBJECTIVE: To study the mechanism of hypoxia inducing factor-1α (HIF-1α) pathway in establishment of hypoxia inducing low endometrial receptivity. METHODS: RL95-2 cell lines, the ideal model of study ER, were cultured in hypoxia condition induced by CoCl2, and the expression of mRNA and protein of HIF-1α and tumor necrosis factor like weak inducer of apoptosis (TWEAK) were measured by reverse transcription-PCR and western blot. The apoptosis rate was analyzed by flow cytometry. Then the mechanism confirmed by comparing the two factors in endometrium and the ultra-appearance of inflammatory reaction and apoptosis between recurrent spontaneous abortion women and control women. RESULTS: (1) On different time point (0, 12, 24, 48 hour), mRNA expression of HIF-1α were 0.272 ± 0.010, 0.354 ± 0.020, 0.591 ± 0.020, 0.890 ± 0.020, while the expression of TWEAK were 0.104 ± 0.010, 0.510 ± 0.020, 1.021 ± 0.020, 1.237 ± 0.040, respectively, the expression level between 12, 24, 48 and 0 hour all showed significant differences (P < 0.05). (2) Protein expression of HIF-1α were 0.853 ± 0.010, 0.931 ± 0.030, 1.124 ± 0.010, 1.317 ± 0.020 respectively, while was 0.042 ± 0.010, 0.091 ± 0.010, 0.131 ± 0.020, 0.205 ± 0.030 in TWEAK expression, the different level were statistically significant (P < 0.05). (3) With longer culture under hypoxia, the cell apoptosis rate increased obviously. The apoptosis rate of each time point were (3.2 ± 1.4)%, (16.2 ± 3.2)%, (26.3 ± 3.5)%, (31.8 ± 3.5)%, the differences between 12, 24, 48 and 0 hour had significance (P < 0.05). (4) The positive rate of HIF-1α stained in epithelium cells and stroma cells of test group were 32.3%, 8.4% and 16.7%, 7.3% in control group. The positive rate of TWEAK were 28.3%, 3.9% in recurrent spontaneous abortion group and 11.6%, 2.7% in control group (P < 0.05). The ultra-appearance of inflammatory cell infiltrated and apoptosis were obvious in test group. CONCLUSIONS: Cell inflammation reaction and apoptosis induced by HIF-1α pathway may participate the mechanism of hypoxia inducing low endometrial receptivity. HIF-1α might become a novel target for improving poor endometrial receptivity.


Asunto(s)
Aborto Habitual/etiología , Apoptosis , Endometrio/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Factores de Necrosis Tumoral/metabolismo , Aborto Habitual/metabolismo , Aborto Habitual/patología , Adulto , Hipoxia de la Célula , Línea Celular Tumoral , Citocina TWEAK , Endometrio/patología , Femenino , Citometría de Flujo , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Embarazo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Tiempo , Factores de Necrosis Tumoral/genética
20.
J Mol Diagn ; 23(6): 710-718, 2021 06.
Artículo en Inglés | MEDLINE | ID: mdl-33781963

RESUMEN

We compared chromosomal mosaicism, detected by next-generation sequencing (NGS), during preimplantation genetic testing (PGT) with that detected by single-nucleotide polymorphism (SNP) array-based PGT to assess the pregnancy outcomes associated with both platforms in a retrospective cohort study of patients undergoing in vitro fertilization in a single university-based assisted reproduction center. In total, 6427 blastocysts biopsied from 1513 patients who underwent 2833 oocyte retrievals from January 2017 to February 2019 were identified. The incidence of mosaicism was significantly higher in the NGS-based PGT group than in the SNP array-based PGT group. Furthermore, some aneuploid specimens were affected by mosaicism. The total mosaicism detection rate with NGS-based PGT (23.3%) was significantly higher than that with SNP array-based PGT (7.7%). Mosaicism rates were similar when stratified by maternal age or PGT type. The SNP array cohort showed a significantly higher spontaneous abortion rate than the NGS cohort (10.07% versus 6.33%; P = 0.0403). The ongoing pregnancy/live birth rate was higher in the NGS cohort (44.1%) than in the SNP array cohort (42.28%). Our results confirm that NGS-based PGT can detect mosaicism more frequently than SNP array-based PGT in trophectoderm specimens. Therefore, clinical application of NGS for PGT may improve pregnancy outcomes compared with that of SNP array-based PGT. More detailed blastocyst detection and classification is necessary to prioritize embryo transfers.


Asunto(s)
Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Mosaicismo , Polimorfismo de Nucleótido Simple , Adulto , Transferencia de Embrión , Femenino , Pruebas Genéticas/métodos , Humanos , Embarazo , Resultado del Embarazo , Estudios Retrospectivos
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