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Objective Using of cumulative live birth rate(CLBR)per oocytes retrieved cycle,to assess the clinical outcomes of in vitro fertilization or intracytoplasmic sperm injection(IVF/ICSI),and to explore impact factors on CLBR following utilization of all fresh and frozen embryos in one complete IVF/ICSI cycle using gonadotropin-releasing hormone(GnRH)agonist, GnRH-antagonist and clomiphene mild stimulation protocols. Methods Of the patients who underwent IVF/ICSI from January 1st, 2014 to December 31st, 2015 in the First Affiliated Hospital, Nanjing Medical University, a total of 6 142 oocytes retrieved cycles were included. The clinical and laboratory parameters of different ovarian stimulation protocols, and the effects of the age, number of oocytes retrieved and number of embryos available on the CLBR of each oocytes retrieved cycle were analyzed.Results The CLBR was 69.0%(2 004/2 906)in the GnRH-agonist protocol versus 67.4%(644/955)in the GnRH-antagonist protocol (P>0.05); the CLBR of clomiphene mild stimulation protocol was 53.2%(1 215/2 281),significantly lower than those of the other two protocols (all P<0.05). The CLBR significantly decreased with age increased. When divided into four groups according to the patients′ age, we found that CLBR were not statistically significant using three different protocols in the 20-25 years old group(all P>0.05).There was a strong association between the number of oocytes retrieved and embryos available on CLBR. CLBR rose significantly with an increasing number of oocytes up to 6, then the rising trend slowed down. Patients were categorized into four groups according to the number of oocytes retrieved,CLBR was significantly higher using GnRH-antagonist protocol (50.0%)than mild stimulation protocol(37.0%)in low ovarian responder(0-4 oocytes)group(P<0.05). The CLBR were no significant difference among three protocols in normal(10-15 oocytes)and high responders(≥15 oocytes)group(all P>0.05).The incidence rate of ovarian hyperstimulation syndrome in GnRH-agonist protocols(5.2%,152/2 906)were significantly higher than those of GnRH-antagonist(4.4%, 42/955)and clomiphene mild stimulation protocols(1.5%,34/2 281;all P<0.05).Conclusions CLBR is an important index to assess the clinical outcomes of IVF/ICSI. Age, number of oocytes retrieved and embryos available could affect CLBR obviously. According to the different age and ovarian response of patients, we should design ovarian stimulation protocols based on target oocytes number in order to get higher CLBR and reduce complications.
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OBJECTIVE To investigate the correlation of 21-hydroxylase deficiency (21-OHD) with male testicular dysplasia. METHODS Clinical data of 8 infertile males with congenital adrenal hyperplasia due to 21-OHD was retrospectively analyzed. In addition, potential mutations of the CYP21A2 gene was detected. RESULTS All patients were referred because of azoospermia or severe oligospermia and had small testis with averaged testicular volume of 6.1 mL. Three patients had testicular adrenal rest tumors. Endocrinologic examinations revealed low levels of leutinizing hormone and follicular stimulating hormone, normal or elevated testosterone, elevated progesterone, elevated or normal adrenocoticotropic hormone, and low or normal cortisol. All patients had adrenal cortical hyperplasia, 5 with adrenal adenoma, 1 case associated with bilateral adrenal myelolipoma. All patients were given glucocorticoid replacement therapy for 3 to 6 months, which successfully improved the seminal status of 6 patient and resulted pregnancies in 5 couples. Seven pathogenic mutations of the CYP21A2 gene among the 8 patients. CONCLUSION 21-OHD can cause testicular hypoplasia and spermatogenic failure. Glucocorticoids and operations can obtain good result and improve spermatogenesis. Our results have shown a good genotype/phenotype correlation in these cases. All patients have carried the p.Ile172Asn mutation, which is associated with simple virilizing form.
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Objective To evaluate the efficiency of the application of array comparative genomic hybridization (array-CGH) in preimplantation genetic diagnosis or screening (PGD/PGS), and compare the clinical outcomes of different stage embryo biopsy. Methods The outcomes of 381 PGD/PGS cycles referred in the First Affiliated Hospital of Nanjing Medical University from July 2011 to August 2015 were retrospectively analyzed. There were 320 PGD cycles with 156 cleavage-stage-biopsy cycles and 164 trophectoderm-biopsy cycles, 61 PGS cycles with 23 cleavage-stage-biopsy cycles and 38 trophectoderm-biopsy cycles.Chromosomal analysis was performed by array-CGH technology combined with whole genome amplification.Single embryo transfer was performed in all transfer cycles.Live birth rate was calculated as the main clinical outcomes. Results The embryo diagnosis rate of PGD/PGS by array-CGH were 96.9%-99.1%. In PGD biopsy cycles, the live birth rate per embryo transfer cycle and live birth rate per embryo biopsy cycle were 50.0%(58/116) and 37.2%(58/156) in cleavage-stage-biopsy group, 67.5%(85/126) and 51.8%(85/164) in trophectoderm-biopsy group (both P<0.01). In PGS biopsy cycles, the live birth rate per embryo transfer cycle and live birth rate per embryo biopsy cycle were the same as 34.8%(8/23) in cleavage-stage-biopsy group, the same as 42.1%(16/38) in trophectoderm-biopsy group (both P>0.05). Conclusions High diagnosis rate and idea live birth rate are achieved in PGD/PGS cycles based on array-CGH technology.The live birth rate of trophectoderm-biopsy group is significantly higher than that of cleavage-stage-biopsy group in PGD cycles;the efficiency of trophectoderm-biopsy is better.
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Objective To study the expression of tight junction factors in human placental tissues derived from assisted reproductive technology (ART) and natural pregnancy and its role in placental barrier.Methods Ten placental samples were collected from the women who had undergone ART treatment and 11 placenta were collected from control group.Transmission electron microscope (TEM) examination was utilized to detect the morphology of placental tight junctions.The mRNA of claudin (CLDN) 1,CLDN4,CLDN5,CLDN8,zonula occudens (ZO) 1 was detected by real-time PCR and the protein of CLDN4,CLDN8 and occludin (OCLN) were measured by western blot.Results TEM microscopy results showed that placenta samples derived both ART and control placenta had normal microscopic histological features of tight junctions,localized in the apical part of the syncytium and also between the cell-cell contacts of fetal blood vessel endothelial.The expression level of CLDN4 mRNA were 0.87 ±0.17 in ART group and 1.18 ± 0.30 in control group,respectively.The expression level of CLDN8 mRNA were 3.25 ± 2.32 in ART group and 1.08±0.41 in control group,respectively.The mRNA level of CLDN4 and CLDN8 were significantly differentially expressed in ART derived placenta when compared with control groups.The expression level of CLDN1,CLDN5,OCLN and ZO1 mRNA were 0.49 ± 0.44,0.80 ± 0.20,0.92 ± 0.18 in ART group and 1.09±0.82,1.21 ±0.78,0.80± 0.27 in control group,respectively,in which there were no significant differences between two groups.Western Blot analysis showed the protein levels of tight junctions CLDN4,CLDN8 and OCLN did not differ between groups.Conclusions Tight junction factors were expressed in human placental tissues.Tight junction derived from ATR platenta might have mild dysfunction.
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Objective Knowledge about the infiltration of macrophages in diabetic wounds can help to figure out the pathogene -sis of poor healing of diabetic wounds .The aim of this study was to observe the macrophage infiltration and the expression of relative in-flammatory factors during the wound healing of diabetic rats and explore the relationship between macrophages and diabetic wound heal -ing. Methods Male Sprague Dawley rats were randomly divided into STZ induced diabetic group and normal control group .Each group had 15 rats.A 1 cm2 full-thickness skin defect was created on the rat dorsum .Wound samples and was excised .On post injury day (PID) 3, 7, and 14, rats of each group were sacrificed after wound samples and tissues around the wound edge were obtained .The differences of the wound closure rate , macrophage infiltration and the relative mRNA expression of the inflammatory factors from macro-phages were observed . Results The wound closure rate was lower in diabetic group on PID 3, 7, and 14 ([29.5 ±5.4]%vs [45.9 ± 12.8]%, [71.6 ±3.1]%vs [80.1 ±6.9]%, [93.9 ±2.8]%vs [99.4 ±1.4]%, P<0.05).HE staining showed inflammatory cells infil-diabetic wound tissue on PID 3 (P<0.05) and a higher expression on PID 14 (P<0.05).The CCR7 fluorescence staining showed more positive staining cells stayed in diabetic wound on PID 14. Conclusion Macrophage infiltration decreases in the early phase of diabetic wound healing and sustains in wound tissue in advanced stage accompanied by the expression change of related inflammatory factors, which could contribute to the difficult wound healing of diabetic rats .
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Objective The gene expression data of T lymphoblastic leukemia cell lines were obtained by " high-throughput sequencing",a new sequence was found,which called J441.The aim of this study was to establish a recombinant lentivirus harboring J441 gene and FLAG-tagged peptide.Methods We got gene expression data from Jurkat by "high-throughput sequencing".A new gene was found by bioinformatic analysis,we called it J441.For further study,a FLAG-J441 fusion gene was constructed by real-time polymerase chain reaction(RT-PCR) and packaged into pHAGE-CMV-MCS-Izs-Green lentivirus vector.The recombinant plasmid pHAGE-J441-FLAG was identified by PCR and sequencing.The recombinant plasmid,packaging vector psPAX2 and envelope vector pMD2.G were co-transfected into 293T cells and the resulting lentivirus was collected.After infection of the recombinant lentivirus,the expression of J441 in 293T was investigated by the expression of ZsGreen and sequencing.The expression of FLAG was investigated by immunofluorescence.Results A recombinant lentivirus plasmid pHAGE-J441-FLAG was successfully constructed.The viral titer was 5 × 1010 ifu/L.The expression of J441 in 293T could be detected by the expression of ZsGreen and sequencing.Immunofluorescence showed that FLAG was expressed in cytoplasm.Conclusions The lentivirus-based delivery system has been successfully constructed.Exogenous J441-FLAG can be stably expressed.The result of immunofluorescence suggest that the J441 gene might be expressed in cytoplasm.
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<p><b>OBJECTIVE</b>To identify potential mutation of ectodysplasin A (EDA) gene in a Chinese family affected with X-linked hypohidrotic ectodermal dysplasia.</p><p><b>METHODS</b>Blood samples were collected from the affected male proband, his family members and 103 unrelated individuals. Following extraction of genomic DNA, coding sequence of the EDA gene was amplified with PCR, and DNA sequencing was performed to detect potential mutation.</p><p><b>RESULTS</b>A novel missense mutation, c.822G>T (p.W274C), was identified in exon 7 of the EDA gene in the proband, whilst his mother was found to be a heterozygous carrier. The same mutation was also found in 5 other family members including one affected male and four females, but was absent in unaffected males and 103 unrelated individuals.</p><p><b>CONCLUSION</b>A c.822G>T mutation in exon 7 of the EDA gene probably underlies the disease in this Chinese family.</p>
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Female , Humans , Male , Young Adult , Asian People , Genetics , Base Sequence , China , Ectodermal Dysplasia 1, Anhidrotic , Diagnosis , Genetics , Ectodysplasins , Genetics , Exons , Mutation , Pedigree , PhenotypeABSTRACT
Objective:To explore the relationship between the polymorphisms in gene FGFR1, FGF10, FGFI8 and the nonsyndromic cleft lip with or without cleft palate (NS CLP) in Chinese population. Methods: Genomic DNA was isolated from peripheral lymphocytes of 75 patients with NS CLP and their parents and 75 unimpaired healthy children. The polymorphisms in FGFRI gene rs13317, p. E467K, p. M3691 and p. S393S, FGF10 gene rs1448037 and FGFI8 gene rs4043716 were detected by applying three-dimensional (3-D) polyacrylamide gel microarray technology. The data were performed using statis-tical analysis : the genotype frequenc+ y and allele frequency between patients with NSCL/P and control subjects were performed. Haplotype relative risk (HRR) , family based association test (FBAT) , and transmission disequilibrium test (TDT) in nuclear family were performed. Results: There were no poly-morphism in FGFR1 gene p. E467K, p. M369I and p. $393S site, the corresponding base was all G. The polymorphisms of rs13317 and rs1448037 were detected and their genotype frequency and allele frequen-cy showed no significant difference between 75 patients with NSCL/P and 75 normal children. TDT, HRR and FBAT were also no significant differences. The genotype frequency of gene FGF18 rs4043716 showed significant difference, but allele frequency were no significant difference. TDT, HRR and FBAT were also no significant difference. Conclusion: Our studies suggest an association between gene FGF18 rs4043716 and the NS CLP in Chinese population, and no association among gene FGFR1 rs13317, p. FA67K, p. M3691, p. S393S and gene FGF10 rs1448037.
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<p><b>OBJECTIVE</b>To assay the expression of neuropeptide Y (NPY) in rat testes and to investigate the significance of NPY in the regulation of androgen production and spermatogenesis.</p><p><b>METHODS</b>NPY mRNA levels in SD rat testes were measured by RT-PCR semi-quantity with beta-actin as internal control. NPY distribution was observed immunohistochemically.</p><p><b>RESULTS</b>NPY gene expressed in the testes, showing the strong positive band in the PCR production electrophoresis gel. In the immunostaining slides, NPY was found positively expressed in the Leyding cell area and around the testicular vessels and tubules, but not in the seminiferous tubules.</p><p><b>CONCLUSION</b>There was positive expression of NPY in the rat testes, which showed that NPY played a direct role in the regulation of testicular function.</p>
Subject(s)
Animals , Male , Rats , Immunohistochemistry , Neuropeptide Y , Genetics , RNA, Messenger , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Testis , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Trx-LHRH, a new GnRH crasis protein, on antibody production and male reproductive function.</p><p><b>METHODS</b>Trx-LHRH produced in vitro with a new crasis gene which crasised Trx gene and GnRH gene together, was used as vaccine, and hydroalaminum base as adjuvant, in adult SD rats. After 5 weeks of the first treatment, the same dosage was used again to enhance the effect of vaccine. Antibody level was measured by ELISA, and androgen level by RIA.</p><p><b>RESULTS</b>Trx-LHRH induced successfully the polycolonal antibody at the level of 1 :1 280 approximately 2 560 after 4 weeks of the first treatment, and 1 : 2 000 after 6 weeks of the enhanced treatment. Testosterone level was reduced significantly (P < 0.01) by ELISA, but there was reasonable variation among individuals. Sperm count was also reduced by Trx-LHRH treatment.</p><p><b>CONCLUSION</b>Trx-LHRH can be used as effective vaccine to induce antibody production, and at the same time, restrain the function of hypothatamas-pituitary-testis axis in vivo.</p>
Subject(s)
Animals , Male , Rats , Antibodies , Blood , Gonadotropin-Releasing Hormone , Allergy and Immunology , Rats, Sprague-Dawley , Recombinant Fusion Proteins , Allergy and Immunology , Sperm Count , Testosterone , Blood , Thioredoxins , Allergy and Immunology , Vaccines, Synthetic , Allergy and ImmunologyABSTRACT
Spermatogenesis, a continuous course of cell proliferation and differentiation, depends on reproductive hormones. FSH and LH-induced T are the main hormones regulating spermatogensis. Intratesticular T is one of the key factors in maintaining spermatogenesis, while FSH is just as important for origination and maintenance of normal spermatogenesis. Sertoli cells are the pivot of hormonal regulation. Interestingly, these reproductive hormones also regulate sperm cell apoptosis spermatogenesis. Further studies on the hormonal regulation of spermatogenesis provide a base for the development of safe and recoverable contraceptives for males.
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Humans , Male , Androgens , Physiology , Apoptosis , Contraception , Follicle Stimulating Hormone , Physiology , Luteinizing Hormone , Physiology , SpermatogenesisABSTRACT
The circulating renin-angiotensin system (RAS) is well known for its role in the maintenance of blood pressure and electrolyte and fluid homeostasis. However, other local angiotensin-generating systems than the circulating RAS have been found in numerous tissues. The male reproductive system including the testis, epididymis, and prostate has several sites of intrinsic RAS activity. The local RAS in these tissues can be responsive to androgens, fat acid, drugs, and hypoxia. There has been evidence for the involvement of the RAS not only in male reproduction, but also in the development of prostate disease. Besides, the assessment of the local RAS activity may be helpful to the early diagnosis of tumor in the male reproductive system.
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Animals , Humans , Male , Rats , Early Diagnosis , Genital Neoplasms, Male , Diagnosis , Genitalia, Male , Metabolism , Physiology , Renin-Angiotensin System , PhysiologyABSTRACT
0.05) after irradiation with low dose of UVB. When the dose of UVB reached 60 mJ/cm2, the mRNA and protein expression of both p21 and Bax increased significantly (both P