Objective:To evaluate whether Ding Kun Dan(DKD)can improve the vitro fertilization/intracytoplasmic sperm injection and embryo transfer(IVF/ICSI-ET)outcomes in patients with predicted poor ovarian response(POR)safely ...Objective:To evaluate whether Ding Kun Dan(DKD)can improve the vitro fertilization/intracytoplasmic sperm injection and embryo transfer(IVF/ICSI-ET)outcomes in patients with predicted poor ovarian response(POR)safely and effectively.Methods:Prospective,multicenter,randomized controlled trial;A total of 278 POR patients were randomized in DKD group or immediate treatment group.Both groups received IVF or ICSI as a standard treatment while in the DKD group,DKD was administrated for 3 months before the IVF/ICSI cycle.The primary outcome was the ongoing pregnancy rate.The secondary outcomes include clinical pregnancy rate,biochemical pregnancy rate,total gonadotropin(Gn)dosage and duration,estradiol(E2)and progesterone(P)levels on human chorionic gonadotropin(hCG)trigger day,cycle cancellation rate,number of oocytes retrieved,high-quality embryo rate and any adverse events.Results:Compared to the immediate IVF treatment group,oral administration of DKD for 3 months before IVF led to a significant increase in ongoing pregnancy rate(30.0%v.s.17.6%,P<0.05),biochemical pregnancy rate(39.2%vs.25.2%,P<0.05),clinical pregnancy rate(36.7%vs.22.7%,P<0.05)and high-quality embryo rate(40.8%vs.32.4%,P<0.05),and a significant decrease in Gn duration(P<0.05).However,no significant differences were found in total dosage of Gn,number of retrieved oocytes,cycle cancellation rate,E2 level and P level on hCG trigger days(P>0.05).No serious adverse events occurred during the intervention period in either group.Conclusion:DKD is a safe and effective intervention to improve the IVF/ICSI-ET outcomes in patients with POR.展开更多
Aim： To investigate the effect of formaldehyde （FA） on testes and the protective effect of vitamin E （VE） against oxidative damage by FA in the testes of adult rats. Methods： Thirty rats were randomly divided in...Aim： To investigate the effect of formaldehyde （FA） on testes and the protective effect of vitamin E （VE） against oxidative damage by FA in the testes of adult rats. Methods： Thirty rats were randomly divided into three groups： （1） control; （2） FA treatment group （FAt）; and （3） FAt ＋ VE group. FAt and FAt ＋ VE groups were exposed to FA by inhalation at a concentration of 10 mg/m^3 for 2 weeks. In addition, FAt ＋ VE group were orally administered VE during the 2-week FA treatment. After the treatment, the histopathological and biochemical changes in testes, as well as the quantity and quality of sperm, were observed. Results： The testicular weight, the quantity and quality of sperm, the activities of superoxide dismutase （SOD）, glutathione peroxidase （GSH-Px） and glutathione （GSH） were significantly decreased whereas the level of malondialdehyde （MDA） was significantly increased in testes of rats in FAt group compared with those in the control group. VE treatment restored these parameters in FAt ＋ VE group. In addition, microscopy with hematoxylin-eosin （HE） staining showed that seminiferous tubules atrophied, seminiferous epithelial cells disintegrated and shed in rats in FAt group and VE treatment significantly improved the testicular structure in FAt ＋ VE group. Conclusion： FA destroys the testicular structure and function in adult rats by inducing oxidative stress, and this damage could be partially reversed by VE.展开更多
Biochemical or clinical changes of hyperandrogenism are important elements of polycystic ovary syndrome （PCOS）. There is currently no consensus on the definition and diagnostic criteria of hyperandrogenism in PCOS. ...Biochemical or clinical changes of hyperandrogenism are important elements of polycystic ovary syndrome （PCOS）. There is currently no consensus on the definition and diagnostic criteria of hyperandrogenism in PCOS. The aim of this study was to investigate the complex symptoms of hyperandrogenic disorders and the correlations between metabolism and hyperandrogenism in patients with PCOS from an outpatient reproductive medicine clinic in China. We conducted a case control study of 125 PCOS patients and 130 controls to evaluate differences in body mass index （BMI）, total testosterone （TT）, modified Ferriman-Gallwey hirsutism score, sex hormone binding globulin （SHBG）, homeostasis model assessment-estimated insulin resistance （HOMA-IR） and free androgen index （FAI） between PCOS patients and controls and subgroups of PCOS. The prevalence of acne and hirsutism did not differ significantly between the hyperandrogenic and non-hyperandrogenic subgroup. Patients with signs of hyper- androgenism had significantly higher BMI （P 〈 0.05）, but differences in TT, SHBG, FAI and waist/hip ratio were insignificant. The odds ratio of overweight was calculated for all PCOS patients. Our results suggest that PCOS patients with high BMI tend to have functional disorders of androgen excess; therefore, BMI may be a strong pre-dictor of hyperandrogenism in PCOS.展开更多
SET is a multifunctional protein involved in regulating many biological processes of the cell cycle. It is also a regulator of steroidogenesis in the ovary. However, the expression of SET protein in testis, and its fu...SET is a multifunctional protein involved in regulating many biological processes of the cell cycle. It is also a regulator of steroidogenesis in the ovary. However, the expression of SET protein in testis, and its function, still remains ambiguous. In this study, we observed the expression of SET in the testes of mice at different developmental stages, and have discussed its potential function in regulating spermatogenesis and androgen production. Forty-eight male mice at different developmental stages （1 week old as the infancy group; 4 weeks old as the prepubertal group; 12 weeks old as the adult group; over 12 months old as the ageing group） were used. Cellular location of SET protein in the testes was observed by immuno-histochemistry. Expression levels of Set mRNA and SET protein were analyzed by quantitative polymerase chain reaction and Western blotting. SET protein was expressed in spermatogonial cells and spermatocytes; the highest level was mainly in haploid and tetraploid cells of the prepubertal and adult groups, and Leydig cells of the adult and ageing groups. There was a low expression in Sertoli cells. Expression of Set mRNA in the prepubertal group was significantly higher than that in the adult group （P 〈 0.05）, while expression of SET protein was at the highest level in the adult group （P 〈 0.05）. SET protein is mainly expressed in spermatogonial cells and spermatocytes, and poorly expressed in Sertoli cells, suggesting that it is involved in spermatogenesis. Expression of SET protein in Leydig cells suggests a possible role in steroidogenesis.展开更多
This study examined the effects of Bangdeyun on the expressions of nuclear factor-kappaB （NF-κB）, interferon-gamma （IFN-y） and interleukin-10 （IL-10） in the endometrium of mice with embryo implantation dysfunct...This study examined the effects of Bangdeyun on the expressions of nuclear factor-kappaB （NF-κB）, interferon-gamma （IFN-y） and interleukin-10 （IL-10） in the endometrium of mice with embryo implantation dysfunction （EID） during the implantation time （namely on pregnancy day 5, 6, 7 and 8） and explored the local immune regulatory effects of Bangdeyun. The gestational mice were randomly divided into normal group, model group and Bangdeyun-treated group. EID models of mice were established by using indomethacin. The endometrial expression of NF-κB was detected by immunohistochemistry and Western blotting. IFN-γ and IL-10 were measured by enzyme-linked immunosorbent assay （ELISA）. The results showed that in the normal group, NF-κB and IFN-γ were weakly expressed and IL-10 was strongly expressed in the endometrium during the whole implantation period. In the model group, the expressions of NF-κB and IFN-T were increased on pregnancy day 5, 6 and 7, and IL-10 expression decreased during the whole implantation time when compared with those in the normal group （P〈0.01 for all）. In the Bangdeyun-treated group, little amount of NF-κB and IFN-γ was expressed and IL-10 expression was strong, much the way they were expressed in the normal group （P〉0.05）. The expressions of NF-κB and IFN-T were much lower in the Bangdeyun-treated group than those in the model group on pregnancy day 5, 6 and 7 （P〈0.01 for all）, while the expression of IL-10 was much higher than in the model group during the whole implantation time （P〈0.01）. It was suggested Bangderun may favor a shift from Thl- to Th2-type immune response, therefore inhibiting the maternal immune rejection, inducing the immune tolerance and improving the fetal implantation.展开更多
Various methods are currently under investigation to preserve fertility in males treated with high-dose chemotherapy and radiation for malignant and nonmalignant disorders. Human umbilical cord mesenchymal stem cells ...Various methods are currently under investigation to preserve fertility in males treated with high-dose chemotherapy and radiation for malignant and nonmalignant disorders. Human umbilical cord mesenchymal stem cells （HUC-MSCs）, which possess potent immunosuppressive function and secrete various cytokines and growth factors, have the potential clinical applications. As a potential alternative, we investigate whether injection of HUC-MSCs into the interstitial compartment of the testes to promote spermatogenic regeneration efficiently. HUC-MSCs were isolated from different sources of umbilical cords and injected into the interstitial space of one testis from 10 busulfan-treated mice （saline and HEK293 cells injections were performed in a separate set of mice） and the other testis remained uninjected. Three weeks after MSCs injection, Relative quantitative reverse transcription polymerase chain reaction was used to identify the expression of 10 of germ cell associated, which are all related to meiosis, demonstrated higher levels of spermatogenic gene expression （2-8 fold） in HUC-MSCs injected testes compared to the contralateral uninjected testes （five mice）. Protein levels for germ cell-specific genes, miwi, vasa and synaptonemal complex protein （Scp3） were also higher in MSC-treated testes compared to injected controls 3 weeks after treatment. However, no different expression was detected in saline water and HEK293 cells injection control group. We have demonstrated HUC-MSCs could affect mouse germ cell-specific genes expression. The results also provide a possibility that the transplanted HUC-MSCs may promote the recovery of spermatogenesis. This study provides further evidence for preclinical therapeutic effects of HUC-MSCs, and explores a new approach to the treatment of azoospermia.展开更多
Objectives:To observe the different expression of CALB2 in the testes of rats at different development stages. Methods:Male SD rats at three development stages were used in this study.Thirty five rats aged 3 to 4 week...Objectives:To observe the different expression of CALB2 in the testes of rats at different development stages. Methods:Male SD rats at three development stages were used in this study.Thirty five rats aged 3 to 4 weeks were as early maturity group,16 rats aged 16 weeks as middle-aged group,and 16 rats aged over 12 months as aged group.Serum testosterone concentration was detected by radioimmunoassay(RIA).Cellular location of CALB2 in testis was observed by immunohistochemistry.Expressions of CALB2 at mRNA and protein level were quantificationally analyzed by qPCR and Western blot. Results:CALB2 was mainly located in cytoplasm of Leydig cells.Serum testosterone levels and expression of CALB2 in both testicular mRNA and protein level were low in the rats of early maturity group.Compared to the early maturity group,serum testosterone levels in the middle-aged group were significantly higher(P<0.01),and testicular expression of CALB2 was significantly increased(P<0.05).Interestingly,the serum testosterone levels and testicular expression of CALB2 in aged group were significantly lower than those in the middle-aged group (P<0.05). Conclusion:calretinin is positively correlated with serum testosterone levels and differently expresses in testes of rats at different development stages,which suggests that testosterone production may be regulated by calretinin as an important paracrine and autocrine factor.展开更多
The human skin has the ability to sense tactile touch and a great range of pressures.Therefore,in prosthetic or robotic systems,it is necessary to prepare pressure sensors with high sensitivity in a wide measurement r...The human skin has the ability to sense tactile touch and a great range of pressures.Therefore,in prosthetic or robotic systems,it is necessary to prepare pressure sensors with high sensitivity in a wide measurement range to provide human-like tactile sensation.Herein,we developed a flexible piezoresistive pressure sensor that is highly sensitive in a broad pressure range by using lotus leaf micropatterned polydimethylsiloxane and multilayer superposition.By superposing four layers of micropatterned constructive substrates,the multilayer piezoresistive pressure sensor achieves a broad pressure range of 312 kPa,a high sensitivity of 2.525 kPa^(−1),a low limit of detection(LOD)of<12 Pa,and a fast response time of 45 ms.Compared with the traditional flexible pressure sensor,the pressure range of this sensor can be increased by at least an order of magnitude.The flexible piezoresistive pressure sensor also shows high robustness:after testing for at least 1000 cycles,it shows no sign of fatigue.More importantly,these sensors can be potentially applied in various human motion detection scenarios,including tiny pulse monitoring,throat vibration detection,and large under-feet pressure sensing.The proposed fabrication strategy may guide the design of other kinds of multifunctional sensors to improve the detection performance.展开更多
Summary： Whether the type of culture media utilized in assisted reproductive technology has impacts on laboratory outcomes and birth weight of newborns in in-vitro fertilization （IVF）/intracytoplasmic sperm injecti...Summary： Whether the type of culture media utilized in assisted reproductive technology has impacts on laboratory outcomes and birth weight of newborns in in-vitro fertilization （IVF）/intracytoplasmic sperm injection （ICSI） was investigated. A total of 673 patients undergoing IVF/ICSI and giving birth to live singletons after fresh embryo transfer on day 3 from Jan. 1, 2010 to Dec. 31, 2012 were included. Three types of culture media were used during this period： Quinn＇s Advantage （QA）, Single Step Medium （SSM）, and Continuous Single Culture medium （CSC）. Fertilization rate （FR）, normal fertilization rate （NFR）, cleavage rate （CR）, normal cleavage rate （NCR）, good-quality embryo rate （GQER） and neonatal birth weight were compared using one-way ANOVA and Z2 tests. Multiple linear regression analysis was performed to determine the impact of culture media on laboratory outcomes and birth weight. In IVF cycles, GQER was significantly decreased in SSM medium group as compared with QA or CSC media groups （63.6% vs. 69.0% in QA; vs. 71.3% in CSC, P=0.011）. In ICSI cycles, FR, NFR and CR were significantly lower in CSC medium group than in other two media groups. No significant difference was observed in neonatal birthweight among the three groups （P=0.759）. Multiple linear regression analyses confirmed that the type of culture medium was correlated with FR, NFR, CR and GQER, but not with neonatal birth weight. The type of culture media had potential influences on laboratory outcomes but did not exhibit an impact on the birth weight of singletons in ART.展开更多
Summary： Human sperm DNA is an important genetic and epigenetic material, whose chromatin structure differs from that of somatic cells. As such, conventional methods for DNA extraction of somatic cells may not be sui...Summary： Human sperm DNA is an important genetic and epigenetic material, whose chromatin structure differs from that of somatic cells. As such, conventional methods for DNA extraction of somatic cells may not be suitable for obtaining sperm DNA. In this study, we evaluated and compared three sperm DNA extraction techniques, namely, modified guanidinium thiocyanate method （method A）, traditional phenol-chloroform method （method B）, and TianGen kit method （method C）. Spectrophotometry and agarose gel electrophoresis analyses showed that method A produced DNA with higher quantity and purity than those of methods B and C （P〈0.01）. PCR results revealed that method A was more reliable in amplifying DEAD-box polypeptide 4 （DDX4） and copy number variations （CNVs） than methods B and C, which generated false-positive errors. The results of sperm DNA methylation assay further indicated that methods A and B were effective, and the former yielded higher quantitative accuracy. In conclusion, the modified guanidinium thiocyanate method provided high quality and reli- able results and could be an optimal technique for extracting sperm DNA for methylation assay.展开更多
The expression of endothelial nitric oxide synthase traffic inducer （NOSTRIN） was examined in the umbilical vessels of the patients with pre-eclampsia （PE） to explore its possible role in the pathogenesis of PE. T...The expression of endothelial nitric oxide synthase traffic inducer （NOSTRIN） was examined in the umbilical vessels of the patients with pre-eclampsia （PE） to explore its possible role in the pathogenesis of PE. The NOSTR1N rnRNA in umbilical tissues was determined by RT-PCR. The eNOS activity in umbilical vessels was spectrophotometrically detected. NO2 /NO3, the stable metabolic end products of NO, was measured by using nitrate reductase. RT-PCR showed that the expression level of NOSTRIN was significantly higher in women with PE than in the normal group （P〈0.01）. The activity of eNOS was significantly decreased in PE group [（12.83±3.61） U/mg] than in normal group [（21.72±3.83） U/mg] （P〈0.01）. The level of NO2-/NO3- in PE patients （27.53±7.48） pmol/mg was significantly lower than that of normal group （54.27±9.53） μmol/mg （P〈0.01）. The significant negative correlation existed between the expression of NOSTRIN and the activity of eNOS in umbilical vessels of women with PE （r=-0.58, P〈0.01）. It was concluded that the level of NOSTR1N expression was increased in umbilical vessel of women with PE, indicating that it may be involved in the pathogenesis of PE.展开更多
Ovarian cancer is the fifth lethal gynecologic malignancy. Metastasis-associated gene 1 （MTA1） is overexpressed in many malignant tumors with high metastatic potential. This study investi- gated whether down-regulat...Ovarian cancer is the fifth lethal gynecologic malignancy. Metastasis-associated gene 1 （MTA1） is overexpressed in many malignant tumors with high metastatic potential. This study investi- gated whether down-regulation of MTA1 expression by RNAi in A2780 ovarian cancer cells could affect proliferation, anoikis, migration, invasion and adhesion of the cells and to research the potential for MTA1 gene therapy of ovarian cancer. After transfection with effective Mtal gene siRNA, the effects on proliferation, anoikis, migration, invasion and adhesion of A2780 cells were tested by MTT assay, flow cytometry, wound-healing assay, Transwell assay and adhesion assay. Expression levels of PTEN, beta 1 integrin, MMP-9, phosphor-AKT （Ser473）, and total AKT activity were evaluated in control and transfected cells. The results showed that inhibition of MTA1 mediated by Mtal-siRNA transfection decreased the cell invasion, migration and adhesion, and induced the increased cell anoikis, but no significant difference was found in proliferation of A2780 cancer cells. In addition, beta 1 integrin, MMP-9, and phosphor-AKT protein levels were significantly down-regulated, while PTEN was significantly up-regulated. These results demonstrated that MTA1 played an important role in the cell metastasis in ovarian cancer. MTA1 could serve as another novel potential therapeutic target in ovarian cancer.展开更多
Summary： Resistance to chemotherapy is a major obstacle for the effective treatment of advanced ovarian cancer. The mechanism of chemoresistance is still poorly understood. Recently, more and more evidence showed mic...Summary： Resistance to chemotherapy is a major obstacle for the effective treatment of advanced ovarian cancer. The mechanism of chemoresistance is still poorly understood. Recently, more and more evidence showed microRNAs （miRNAs） modulated many key molecules and pathways involved in chemotherapy, microRNA-106a （miR-106a） has been implicated in many cancers, but its role in ovarian cancer and drug resistance still remains unexplored. This study was to investigate whether miR-106a mediated resistance of the ovarian cancer cell line A2780 to the chemotherapeutic agent cisplatin （DDP）. The different levels of miR-106a in A2780 cells and their resistant variant A2780/DDP cells were identified by using real-time PCR. MTT assay and flow cytometry were used to analyze the effect of miR-106a on cisplatin resistance of these paired cells. Real-time PCR, Western blotting and luciferase reporter assay were applied to explore whether Mcl-1 was a target of miR-106a. As compared to A2780 cells, the expression of miR-106a was down-regulated in the cisplatin resistant cell line A2780/DDP. Moreover, knockdown of miR-106a dramatically decreased antiproliferative effects and apoptosis induced by cisplatin in A2780 cells, while overexpression of miR-106a significantly increased antiproliferative effects and apoptosis induced by cisplatin in A2780/DDP cells. Furthermore, miR-106a inhibited cell survival and cisplatin resistance through downregulating the expression of Mcl-1. Mcl-1 was a di- rect target of miR-106a. These results suggest that miR-106a may provide a novel mechanism for un- derstanding cisplatin resistance in ovarian cancer by modulating Mcl-1.展开更多
Objective To investigate the expression of routine cytomegalovirus （MCMV） M83 mRNA in mouse testes at the different periods of MCMV infection and its relationship with the severity of testicular lesions Methods Used...Objective To investigate the expression of routine cytomegalovirus （MCMV） M83 mRNA in mouse testes at the different periods of MCMV infection and its relationship with the severity of testicular lesions Methods Used digoxin-labeled MCMV mRNA oligonucleotide probe, the mouse testicular tissues at the different periods following MCMV inoculation were detected by in situ hybridization（ISH）. The average absorbency （A） of ISH positive signals represented the resticular MCMV load. The expression of MCMV M83 mRNA in the infected testes and its relation with the severity of testicular lesions were analyzed. Results At different periods after MCMV inoculation, the expression of MCMV M83 mRNA was detectable in the testicular tissues. The ISH positive signals were mainly located in cytoplasm of the testicular interstitial cells. The average A values at d 9 after MCMV infection were the highest （comparison between the experimental groups, P〈O. O01）. There were pathological inJTammatory changes to varying degrees in seminiferous epithelia and interstitial cells at the diJferent periods of infection, especially during d 6 to d 9. Conclusion In testicular MCMV infection, the levels of MCMV M83 mRNA were related with the severity of testicular lesions, suggesting the dynamic detection of MCMV M83 mRNA could evaluate the prognosis of MCMV infection.展开更多
Objective： To study the effects of simulated microgravity induced by tail-suspension on histological structures and antioxidant system in testis of adult rats. Methods： Forty Spraque-Dawley adult male rats were rand...Objective： To study the effects of simulated microgravity induced by tail-suspension on histological structures and antioxidant system in testis of adult rats. Methods： Forty Spraque-Dawley adult male rats were randomly divided into four groups, two experimental groups and two synchronic control groups. Rats in the two experimental groups were tail-suspended for 14 d and 28 d respectively, then serum hormone, testicular morphology and biochemical changes were evaluated. Results： Compared with the synchronic controls, serum testosterone （T） levels and testicular superoxide dismutase （SOD） significantly decreased（P 〈 0.05）, while testicular malondialdehyde（MDA） significantly increased （P 〈 0.05） in tail-suspended rats. The changes were more prominent in the 28 d tail-suspended rats. In addition, hematoxylin and eosin （HE） staining showed that seminiferous tubules atrophied, spermatogenic cells decreased, and seminiferous epithelial cells disintegrated and shed, abnormal multinucleated giant cells appeared and the lumina were azoospermic in tail-suspended rats. ConcIusion： Simulated microgravity induced by tailsuspension has a harmful effect on male reproduction of adult rats by decreasing the ability of testicular antioxidant defense.展开更多
This study examined the expression and distribution of angiopoietin-1/-2 （Ang-1/-2） in the endometrium of early pregnant mice. The expression of Ang-1/-2 was detected by immunohistochemical staining and in situ hybr...This study examined the expression and distribution of angiopoietin-1/-2 （Ang-1/-2） in the endometrium of early pregnant mice. The expression of Ang-1/-2 was detected by immunohistochemical staining and in situ hybridization respectively. Computerized image analysis system was used to measure the average optical intensity of Ang-1/-2 in endometria at different time points after gestation. Mice were randomly divided into 5 groups： control group, D2 group （2 days after pregnancy）, D4 group （4 days after pregnancy）, D6 group （6 days after pregnancy） and D8 group （8 days after pregnancy）, each containing 15 mice. The results showed that the expression of Ang-1 and Ang-2 was very different among 4 groups （P〈0.01）. Immunohistochemical staining revealed that Ang-1 was localized in the cytoplasma of stromal cells 2 days after pregnancy （day 2）, and in luminal epithelial cells on day 4. The protein of Ang-2 was mainly expressed in the cytoplasma of glandular epithelia and stromal cells. With gestation time, the positive reactions of Ang-1/-2 were stronger in the endometria of the pregnant mice （P〈0.01）. In situ hybridization showed Ang-I mRNA in stromal cells on day 2. Hybridization signal was localized in both stromal cells and vessel epithelial cells on day 4; Ang-2 mRNA was expressed in stromal cells and glandular epithelia on day 2; high mRNA levels appeared in stromal cells, glandular epithelia and vascular endothelia on day 4; an increasing in mRNA expression of Ang-1/-2 was observed on day 6 and day 8 （P〈0.01）. It is suggested that Ang-1/-2 may play an important role in the cross-talk between blastocyst and maternal endometrium during the process of embryo implantation.展开更多
Brain-derived neurotrophic factor (BDNF) can promote developmental competence in mammalian oocytes during in vitro maturation (IVM),but the role of BDNF in oocyte maturation at cellular level is not still clear.In thi...Brain-derived neurotrophic factor (BDNF) can promote developmental competence in mammalian oocytes during in vitro maturation (IVM),but the role of BDNF in oocyte maturation at cellular level is not still clear.In this study,mouse cumulus-enclosed oocytes subjected to IVM were fertilized and cultured to blastocyst stage.Meiotic spindle configuration and cortical granules distribution during oocyte maturation in vitro were assessed by using immunofluorescence and laser confocal microscopy.The results showed that BDNF contributed to the complete preimplantation development of mouse oocytes compared to the control oocytes (13.78% vs.5.92%;P【0.05).Further,BDNF did not accelerate nuclear maturation of IVM oocytes.For the BDNF-treated oocytes at meiosis Ⅰ,Meiotic spindle areas were significantly smaller and the number of cytoplasmic microtubule organizing centers was greater than that in the control,and the percentages of oocytes showed spindles positioned near the oolemma and a well-formed cortical granule-free domain were significantly higher than that of the control.These morphological characteristics of the BDNF-treated oocytes were much closer to the oocytes matured in vivo than those of the control oocytes.In conclusion,BDNF can promote the developmental competence of mouse IVM oocytes,by improving the meiotic spindle configuration and location and cortical granules distribution at meiosis Ⅰ.展开更多
Objective To explore the role of urokinase-type plasminogen activator（uPA） in precontact sperm-egg communication and fertility of mice in vitro. Methods Firstly, sperm chemotaxis （SC） induced by uPA was assayed by...Objective To explore the role of urokinase-type plasminogen activator（uPA） in precontact sperm-egg communication and fertility of mice in vitro. Methods Firstly, sperm chemotaxis （SC） induced by uPA was assayed by measuring the sperm densities in capillaries with a descending gradient or no gradient of uPA respectively. Secondly, the role of uPAR that exists in sperm plasma membrane in SC was studied by examining the change of sperm density in capillary after incubating spermatozoa with anti-uPAR antibody. Thirdly, SC induced by eggs, which had been treated with uPA, PAl-1 and anti-uPAR beforehand respectively, was assayed to study the role of uPA in PSEC. Lastly, the fertilization capability of spermatozoa treated with uPA was examined by counting the number of fertilized eggs. Results 1）The density of spermatozoa that migrated down the gradient of uPA into the capillary was significantly lower than that into the capillary containing no-gradient uPA. 2） When uPAR of spermatozoa was inhibited by anti-uPAR antibody, the density of spermatozoa that migrated into the capillary with ascending gradient of uPA decreased correspondingly. 3） The density of spermatozoa attracted by eggs, which were treated with uPA beforehand, increased significantly than that of attracted by non-treated eggs. On the contrary, the sperm density decreased correspondingly when the egg was treated with PAI-1. 4） The number of fertilized eggs increased significantly after the spermatozoa used here was treated with uPA beforehand. Conclusion uPA could induce SC of mice sperm in vitro through the uPAR on its membrane, enhance the capability of egg inducing SC, and promote spermatozoa to fertilize eggs. Thus, uPA may act as an attractant in PSEC, increase the chance encounter of spermatozoa and eggs, therefore, enhance the fertility success correspondingly. This study, in some degree, provides an evidence that uPA may be used as a new medicine and diagnostic reagent for male infertility.展开更多
In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor （LIF） in mice with embryonic implantation dysfunction （EID）, 120 Kunming mice post coition were rand...In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor （LIF） in mice with embryonic implantation dysfunction （EID）, 120 Kunming mice post coition were randomized into three groups： normal control group, model group and traditional Chinese medicine group （TCM group） （n=40 in each group）. Uterus was collected on the pregnancy day （Pd） 4, 5, 6 after an intravenous injection of Evan＇s blue. The endometrium was dyed by Evan＇s blue and the mean points of response were observed on Pd 5. The expression of LIF mRNA and protein was detected by RT-PCR and immunohistochemistry respectively and analyzed statistically by image system. The results showed that the number of implantation sites in model group was remarkably less than in normal control group and TCM group. There was no significant difference between normal control group and TCM group. The expression of LIF mRNA and protein in model group was delayed. Bushenantai recipe could increase the expression of LIF mRNA and protein in endometria of mice with EID. It was suggested that Bushenantai recipe could improve embryo implantation of mice with EID by promoting the endometrial LIF expression and endometrial decidualization.展开更多
There were few studies of cumulative live birth rates(CLBRs) based on multicenter reproductive clinical data from the general Chinese population.Here we report a retrospective cohort study,including 14 311 women with1...There were few studies of cumulative live birth rates(CLBRs) based on multicenter reproductive clinical data from the general Chinese population.Here we report a retrospective cohort study,including 14 311 women with17 315 cycles,in three reproductive centers to evaluate two estimated parameters of CLBRs with multiple transfer cycles of in vitro fertilization(IVF)/intracytoplasmic sperm injection(ICSI) in a Chinese population.We found that CLBRs were related to female age and endometrial thickness.By the fourth transfer cycle,the conservative and optimal estimates of CLBRs were 52.95% and 77.30% in women under 30 years of age,and 18.17% and26.51% in those 37 years of age or older,respectively.The two estimates were 44.70% and 63.15% in women with endometrial thickness more than 7 mm,and 32.05% and 46.18% in those with less than 7 mm,respectively.In addition,body mass index(BMI),duration of infertility,and infertility diagnoses may also be related to CLBRs on certain conditions.The findings from this study on CLBRs after multiple transfer cycles of IVF/ICSI treatment on different conditions in the Chinese population should be beneficial to both infertile couples and clinicians.展开更多
基金This study was supported by the National Natural Science Foundation of China Nos.81774351the Tianjin Health&Family Planning Commission(Tianjin scientific research project of priority area of traditional Chinese medicine,Nos.2018008)Shanxi Guangyuyuan TCM Co.,Ltd.
文摘Objective:To evaluate whether Ding Kun Dan(DKD)can improve the vitro fertilization/intracytoplasmic sperm injection and embryo transfer(IVF/ICSI-ET)outcomes in patients with predicted poor ovarian response(POR)safely and effectively.Methods:Prospective,multicenter,randomized controlled trial;A total of 278 POR patients were randomized in DKD group or immediate treatment group.Both groups received IVF or ICSI as a standard treatment while in the DKD group,DKD was administrated for 3 months before the IVF/ICSI cycle.The primary outcome was the ongoing pregnancy rate.The secondary outcomes include clinical pregnancy rate,biochemical pregnancy rate,total gonadotropin(Gn)dosage and duration,estradiol(E2)and progesterone(P)levels on human chorionic gonadotropin(hCG)trigger day,cycle cancellation rate,number of oocytes retrieved,high-quality embryo rate and any adverse events.Results:Compared to the immediate IVF treatment group,oral administration of DKD for 3 months before IVF led to a significant increase in ongoing pregnancy rate(30.0%v.s.17.6%,P<0.05),biochemical pregnancy rate(39.2%vs.25.2%,P<0.05),clinical pregnancy rate(36.7%vs.22.7%,P<0.05)and high-quality embryo rate(40.8%vs.32.4%,P<0.05),and a significant decrease in Gn duration(P<0.05).However,no significant differences were found in total dosage of Gn,number of retrieved oocytes,cycle cancellation rate,E2 level and P level on hCG trigger days(P>0.05).No serious adverse events occurred during the intervention period in either group.Conclusion:DKD is a safe and effective intervention to improve the IVF/ICSI-ET outcomes in patients with POR.
基金grants from the SciTechnical Development Project of Shaanxi Province, China （2005K15-G2） Traditional Chinese Project of Shaanxi Province, China （2005024） Natural Science Foundation of Xi＇ an Jiaotong University （573026）.
文摘Aim： To investigate the effect of formaldehyde （FA） on testes and the protective effect of vitamin E （VE） against oxidative damage by FA in the testes of adult rats. Methods： Thirty rats were randomly divided into three groups： （1） control; （2） FA treatment group （FAt）; and （3） FAt ＋ VE group. FAt and FAt ＋ VE groups were exposed to FA by inhalation at a concentration of 10 mg/m^3 for 2 weeks. In addition, FAt ＋ VE group were orally administered VE during the 2-week FA treatment. After the treatment, the histopathological and biochemical changes in testes, as well as the quantity and quality of sperm, were observed. Results： The testicular weight, the quantity and quality of sperm, the activities of superoxide dismutase （SOD）, glutathione peroxidase （GSH-Px） and glutathione （GSH） were significantly decreased whereas the level of malondialdehyde （MDA） was significantly increased in testes of rats in FAt group compared with those in the control group. VE treatment restored these parameters in FAt ＋ VE group. In addition, microscopy with hematoxylin-eosin （HE） staining showed that seminiferous tubules atrophied, seminiferous epithelial cells disintegrated and shed in rats in FAt group and VE treatment significantly improved the testicular structure in FAt ＋ VE group. Conclusion： FA destroys the testicular structure and function in adult rats by inducing oxidative stress, and this damage could be partially reversed by VE.
基金supported by grants from the Major State Basic Research Development Program of China(973 Program:No.2012CB944902 and No.2012CB944703)the National Natural Science Foundation of China(No.30801236)the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘Biochemical or clinical changes of hyperandrogenism are important elements of polycystic ovary syndrome （PCOS）. There is currently no consensus on the definition and diagnostic criteria of hyperandrogenism in PCOS. The aim of this study was to investigate the complex symptoms of hyperandrogenic disorders and the correlations between metabolism and hyperandrogenism in patients with PCOS from an outpatient reproductive medicine clinic in China. We conducted a case control study of 125 PCOS patients and 130 controls to evaluate differences in body mass index （BMI）, total testosterone （TT）, modified Ferriman-Gallwey hirsutism score, sex hormone binding globulin （SHBG）, homeostasis model assessment-estimated insulin resistance （HOMA-IR） and free androgen index （FAI） between PCOS patients and controls and subgroups of PCOS. The prevalence of acne and hirsutism did not differ significantly between the hyperandrogenic and non-hyperandrogenic subgroup. Patients with signs of hyper- androgenism had significantly higher BMI （P 〈 0.05）, but differences in TT, SHBG, FAI and waist/hip ratio were insignificant. The odds ratio of overweight was calculated for all PCOS patients. Our results suggest that PCOS patients with high BMI tend to have functional disorders of androgen excess; therefore, BMI may be a strong pre-dictor of hyperandrogenism in PCOS.
文摘SET is a multifunctional protein involved in regulating many biological processes of the cell cycle. It is also a regulator of steroidogenesis in the ovary. However, the expression of SET protein in testis, and its function, still remains ambiguous. In this study, we observed the expression of SET in the testes of mice at different developmental stages, and have discussed its potential function in regulating spermatogenesis and androgen production. Forty-eight male mice at different developmental stages （1 week old as the infancy group; 4 weeks old as the prepubertal group; 12 weeks old as the adult group; over 12 months old as the ageing group） were used. Cellular location of SET protein in the testes was observed by immuno-histochemistry. Expression levels of Set mRNA and SET protein were analyzed by quantitative polymerase chain reaction and Western blotting. SET protein was expressed in spermatogonial cells and spermatocytes; the highest level was mainly in haploid and tetraploid cells of the prepubertal and adult groups, and Leydig cells of the adult and ageing groups. There was a low expression in Sertoli cells. Expression of Set mRNA in the prepubertal group was significantly higher than that in the adult group （P 〈 0.05）, while expression of SET protein was at the highest level in the adult group （P 〈 0.05）. SET protein is mainly expressed in spermatogonial cells and spermatocytes, and poorly expressed in Sertoli cells, suggesting that it is involved in spermatogenesis. Expression of SET protein in Leydig cells suggests a possible role in steroidogenesis.
基金supported by a grant from National Natural Science Foundation of China (No. 30801538)
文摘This study examined the effects of Bangdeyun on the expressions of nuclear factor-kappaB （NF-κB）, interferon-gamma （IFN-y） and interleukin-10 （IL-10） in the endometrium of mice with embryo implantation dysfunction （EID） during the implantation time （namely on pregnancy day 5, 6, 7 and 8） and explored the local immune regulatory effects of Bangdeyun. The gestational mice were randomly divided into normal group, model group and Bangdeyun-treated group. EID models of mice were established by using indomethacin. The endometrial expression of NF-κB was detected by immunohistochemistry and Western blotting. IFN-γ and IL-10 were measured by enzyme-linked immunosorbent assay （ELISA）. The results showed that in the normal group, NF-κB and IFN-γ were weakly expressed and IL-10 was strongly expressed in the endometrium during the whole implantation period. In the model group, the expressions of NF-κB and IFN-T were increased on pregnancy day 5, 6 and 7, and IL-10 expression decreased during the whole implantation time when compared with those in the normal group （P〈0.01 for all）. In the Bangdeyun-treated group, little amount of NF-κB and IFN-γ was expressed and IL-10 expression was strong, much the way they were expressed in the normal group （P〉0.05）. The expressions of NF-κB and IFN-T were much lower in the Bangdeyun-treated group than those in the model group on pregnancy day 5, 6 and 7 （P〈0.01 for all）, while the expression of IL-10 was much higher than in the model group during the whole implantation time （P〈0.01）. It was suggested Bangderun may favor a shift from Thl- to Th2-type immune response, therefore inhibiting the maternal immune rejection, inducing the immune tolerance and improving the fetal implantation.
文摘Various methods are currently under investigation to preserve fertility in males treated with high-dose chemotherapy and radiation for malignant and nonmalignant disorders. Human umbilical cord mesenchymal stem cells （HUC-MSCs）, which possess potent immunosuppressive function and secrete various cytokines and growth factors, have the potential clinical applications. As a potential alternative, we investigate whether injection of HUC-MSCs into the interstitial compartment of the testes to promote spermatogenic regeneration efficiently. HUC-MSCs were isolated from different sources of umbilical cords and injected into the interstitial space of one testis from 10 busulfan-treated mice （saline and HEK293 cells injections were performed in a separate set of mice） and the other testis remained uninjected. Three weeks after MSCs injection, Relative quantitative reverse transcription polymerase chain reaction was used to identify the expression of 10 of germ cell associated, which are all related to meiosis, demonstrated higher levels of spermatogenic gene expression （2-8 fold） in HUC-MSCs injected testes compared to the contralateral uninjected testes （five mice）. Protein levels for germ cell-specific genes, miwi, vasa and synaptonemal complex protein （Scp3） were also higher in MSC-treated testes compared to injected controls 3 weeks after treatment. However, no different expression was detected in saline water and HEK293 cells injection control group. We have demonstrated HUC-MSCs could affect mouse germ cell-specific genes expression. The results also provide a possibility that the transplanted HUC-MSCs may promote the recovery of spermatogenesis. This study provides further evidence for preclinical therapeutic effects of HUC-MSCs, and explores a new approach to the treatment of azoospermia.
基金supported by projects from China National Nature Foundation(81170559)Jiangsu Health Foundation(XK02200901-NG09)
文摘Objectives:To observe the different expression of CALB2 in the testes of rats at different development stages. Methods:Male SD rats at three development stages were used in this study.Thirty five rats aged 3 to 4 weeks were as early maturity group,16 rats aged 16 weeks as middle-aged group,and 16 rats aged over 12 months as aged group.Serum testosterone concentration was detected by radioimmunoassay(RIA).Cellular location of CALB2 in testis was observed by immunohistochemistry.Expressions of CALB2 at mRNA and protein level were quantificationally analyzed by qPCR and Western blot. Results:CALB2 was mainly located in cytoplasm of Leydig cells.Serum testosterone levels and expression of CALB2 in both testicular mRNA and protein level were low in the rats of early maturity group.Compared to the early maturity group,serum testosterone levels in the middle-aged group were significantly higher(P<0.01),and testicular expression of CALB2 was significantly increased(P<0.05).Interestingly,the serum testosterone levels and testicular expression of CALB2 in aged group were significantly lower than those in the middle-aged group (P<0.05). Conclusion:calretinin is positively correlated with serum testosterone levels and differently expresses in testes of rats at different development stages,which suggests that testosterone production may be regulated by calretinin as an important paracrine and autocrine factor.
基金the Project of National Key Research and Development Program of China(No.2018YFC2001300)the National Natural Science Foundation of China(Nos.52175271,51822504,52021003,52105299,51905207,and 91948302)+2 种基金Science and Technology Development Plan Project of Jilin Province(No.20210508057RQ)Program for JinlinUniversity Science and Technology Innovative Research Team(No.2017TD-04)Scientific Research Project of EducationDepartment of Jilin Province(No.JJKH20211084KJ).
文摘The human skin has the ability to sense tactile touch and a great range of pressures.Therefore,in prosthetic or robotic systems,it is necessary to prepare pressure sensors with high sensitivity in a wide measurement range to provide human-like tactile sensation.Herein,we developed a flexible piezoresistive pressure sensor that is highly sensitive in a broad pressure range by using lotus leaf micropatterned polydimethylsiloxane and multilayer superposition.By superposing four layers of micropatterned constructive substrates,the multilayer piezoresistive pressure sensor achieves a broad pressure range of 312 kPa,a high sensitivity of 2.525 kPa^(−1),a low limit of detection(LOD)of<12 Pa,and a fast response time of 45 ms.Compared with the traditional flexible pressure sensor,the pressure range of this sensor can be increased by at least an order of magnitude.The flexible piezoresistive pressure sensor also shows high robustness:after testing for at least 1000 cycles,it shows no sign of fatigue.More importantly,these sensors can be potentially applied in various human motion detection scenarios,including tiny pulse monitoring,throat vibration detection,and large under-feet pressure sensing.The proposed fabrication strategy may guide the design of other kinds of multifunctional sensors to improve the detection performance.
文摘Summary： Whether the type of culture media utilized in assisted reproductive technology has impacts on laboratory outcomes and birth weight of newborns in in-vitro fertilization （IVF）/intracytoplasmic sperm injection （ICSI） was investigated. A total of 673 patients undergoing IVF/ICSI and giving birth to live singletons after fresh embryo transfer on day 3 from Jan. 1, 2010 to Dec. 31, 2012 were included. Three types of culture media were used during this period： Quinn＇s Advantage （QA）, Single Step Medium （SSM）, and Continuous Single Culture medium （CSC）. Fertilization rate （FR）, normal fertilization rate （NFR）, cleavage rate （CR）, normal cleavage rate （NCR）, good-quality embryo rate （GQER） and neonatal birth weight were compared using one-way ANOVA and Z2 tests. Multiple linear regression analysis was performed to determine the impact of culture media on laboratory outcomes and birth weight. In IVF cycles, GQER was significantly decreased in SSM medium group as compared with QA or CSC media groups （63.6% vs. 69.0% in QA; vs. 71.3% in CSC, P=0.011）. In ICSI cycles, FR, NFR and CR were significantly lower in CSC medium group than in other two media groups. No significant difference was observed in neonatal birthweight among the three groups （P=0.759）. Multiple linear regression analyses confirmed that the type of culture medium was correlated with FR, NFR, CR and GQER, but not with neonatal birth weight. The type of culture media had potential influences on laboratory outcomes but did not exhibit an impact on the birth weight of singletons in ART.
基金supported by the National Natural Science Foundation of China(No.81370755)
文摘Summary： Human sperm DNA is an important genetic and epigenetic material, whose chromatin structure differs from that of somatic cells. As such, conventional methods for DNA extraction of somatic cells may not be suitable for obtaining sperm DNA. In this study, we evaluated and compared three sperm DNA extraction techniques, namely, modified guanidinium thiocyanate method （method A）, traditional phenol-chloroform method （method B）, and TianGen kit method （method C）. Spectrophotometry and agarose gel electrophoresis analyses showed that method A produced DNA with higher quantity and purity than those of methods B and C （P〈0.01）. PCR results revealed that method A was more reliable in amplifying DEAD-box polypeptide 4 （DDX4） and copy number variations （CNVs） than methods B and C, which generated false-positive errors. The results of sperm DNA methylation assay further indicated that methods A and B were effective, and the former yielded higher quantitative accuracy. In conclusion, the modified guanidinium thiocyanate method provided high quality and reli- able results and could be an optimal technique for extracting sperm DNA for methylation assay.
文摘The expression of endothelial nitric oxide synthase traffic inducer （NOSTRIN） was examined in the umbilical vessels of the patients with pre-eclampsia （PE） to explore its possible role in the pathogenesis of PE. The NOSTR1N rnRNA in umbilical tissues was determined by RT-PCR. The eNOS activity in umbilical vessels was spectrophotometrically detected. NO2 /NO3, the stable metabolic end products of NO, was measured by using nitrate reductase. RT-PCR showed that the expression level of NOSTRIN was significantly higher in women with PE than in the normal group （P〈0.01）. The activity of eNOS was significantly decreased in PE group [（12.83±3.61） U/mg] than in normal group [（21.72±3.83） U/mg] （P〈0.01）. The level of NO2-/NO3- in PE patients （27.53±7.48） pmol/mg was significantly lower than that of normal group （54.27±9.53） μmol/mg （P〈0.01）. The significant negative correlation existed between the expression of NOSTRIN and the activity of eNOS in umbilical vessels of women with PE （r=-0.58, P〈0.01）. It was concluded that the level of NOSTR1N expression was increased in umbilical vessel of women with PE, indicating that it may be involved in the pathogenesis of PE.
基金supported by the youth innovation fundation of the First Affiliated Hospital of Zhengzhou University
文摘Ovarian cancer is the fifth lethal gynecologic malignancy. Metastasis-associated gene 1 （MTA1） is overexpressed in many malignant tumors with high metastatic potential. This study investi- gated whether down-regulation of MTA1 expression by RNAi in A2780 ovarian cancer cells could affect proliferation, anoikis, migration, invasion and adhesion of the cells and to research the potential for MTA1 gene therapy of ovarian cancer. After transfection with effective Mtal gene siRNA, the effects on proliferation, anoikis, migration, invasion and adhesion of A2780 cells were tested by MTT assay, flow cytometry, wound-healing assay, Transwell assay and adhesion assay. Expression levels of PTEN, beta 1 integrin, MMP-9, phosphor-AKT （Ser473）, and total AKT activity were evaluated in control and transfected cells. The results showed that inhibition of MTA1 mediated by Mtal-siRNA transfection decreased the cell invasion, migration and adhesion, and induced the increased cell anoikis, but no significant difference was found in proliferation of A2780 cancer cells. In addition, beta 1 integrin, MMP-9, and phosphor-AKT protein levels were significantly down-regulated, while PTEN was significantly up-regulated. These results demonstrated that MTA1 played an important role in the cell metastasis in ovarian cancer. MTA1 could serve as another novel potential therapeutic target in ovarian cancer.
基金supported by the Youth Innovation Foundation of the First Affiliated Hospital of Zhengzhou University
文摘Summary： Resistance to chemotherapy is a major obstacle for the effective treatment of advanced ovarian cancer. The mechanism of chemoresistance is still poorly understood. Recently, more and more evidence showed microRNAs （miRNAs） modulated many key molecules and pathways involved in chemotherapy, microRNA-106a （miR-106a） has been implicated in many cancers, but its role in ovarian cancer and drug resistance still remains unexplored. This study was to investigate whether miR-106a mediated resistance of the ovarian cancer cell line A2780 to the chemotherapeutic agent cisplatin （DDP）. The different levels of miR-106a in A2780 cells and their resistant variant A2780/DDP cells were identified by using real-time PCR. MTT assay and flow cytometry were used to analyze the effect of miR-106a on cisplatin resistance of these paired cells. Real-time PCR, Western blotting and luciferase reporter assay were applied to explore whether Mcl-1 was a target of miR-106a. As compared to A2780 cells, the expression of miR-106a was down-regulated in the cisplatin resistant cell line A2780/DDP. Moreover, knockdown of miR-106a dramatically decreased antiproliferative effects and apoptosis induced by cisplatin in A2780 cells, while overexpression of miR-106a significantly increased antiproliferative effects and apoptosis induced by cisplatin in A2780/DDP cells. Furthermore, miR-106a inhibited cell survival and cisplatin resistance through downregulating the expression of Mcl-1. Mcl-1 was a di- rect target of miR-106a. These results suggest that miR-106a may provide a novel mechanism for un- derstanding cisplatin resistance in ovarian cancer by modulating Mcl-1.
文摘Objective To investigate the expression of routine cytomegalovirus （MCMV） M83 mRNA in mouse testes at the different periods of MCMV infection and its relationship with the severity of testicular lesions Methods Used digoxin-labeled MCMV mRNA oligonucleotide probe, the mouse testicular tissues at the different periods following MCMV inoculation were detected by in situ hybridization（ISH）. The average absorbency （A） of ISH positive signals represented the resticular MCMV load. The expression of MCMV M83 mRNA in the infected testes and its relation with the severity of testicular lesions were analyzed. Results At different periods after MCMV inoculation, the expression of MCMV M83 mRNA was detectable in the testicular tissues. The ISH positive signals were mainly located in cytoplasm of the testicular interstitial cells. The average A values at d 9 after MCMV infection were the highest （comparison between the experimental groups, P〈O. O01）. There were pathological inJTammatory changes to varying degrees in seminiferous epithelia and interstitial cells at the diJferent periods of infection, especially during d 6 to d 9. Conclusion In testicular MCMV infection, the levels of MCMV M83 mRNA were related with the severity of testicular lesions, suggesting the dynamic detection of MCMV M83 mRNA could evaluate the prognosis of MCMV infection.
基金Scientific Technology Project Foundation of Shaanxi Province(2005K15-G2)and Scientific Research Foundation of Xi’an Jiaotong University(573026)
文摘Objective： To study the effects of simulated microgravity induced by tail-suspension on histological structures and antioxidant system in testis of adult rats. Methods： Forty Spraque-Dawley adult male rats were randomly divided into four groups, two experimental groups and two synchronic control groups. Rats in the two experimental groups were tail-suspended for 14 d and 28 d respectively, then serum hormone, testicular morphology and biochemical changes were evaluated. Results： Compared with the synchronic controls, serum testosterone （T） levels and testicular superoxide dismutase （SOD） significantly decreased（P 〈 0.05）, while testicular malondialdehyde（MDA） significantly increased （P 〈 0.05） in tail-suspended rats. The changes were more prominent in the 28 d tail-suspended rats. In addition, hematoxylin and eosin （HE） staining showed that seminiferous tubules atrophied, spermatogenic cells decreased, and seminiferous epithelial cells disintegrated and shed, abnormal multinucleated giant cells appeared and the lumina were azoospermic in tail-suspended rats. ConcIusion： Simulated microgravity induced by tailsuspension has a harmful effect on male reproduction of adult rats by decreasing the ability of testicular antioxidant defense.
文摘This study examined the expression and distribution of angiopoietin-1/-2 （Ang-1/-2） in the endometrium of early pregnant mice. The expression of Ang-1/-2 was detected by immunohistochemical staining and in situ hybridization respectively. Computerized image analysis system was used to measure the average optical intensity of Ang-1/-2 in endometria at different time points after gestation. Mice were randomly divided into 5 groups： control group, D2 group （2 days after pregnancy）, D4 group （4 days after pregnancy）, D6 group （6 days after pregnancy） and D8 group （8 days after pregnancy）, each containing 15 mice. The results showed that the expression of Ang-1 and Ang-2 was very different among 4 groups （P〈0.01）. Immunohistochemical staining revealed that Ang-1 was localized in the cytoplasma of stromal cells 2 days after pregnancy （day 2）, and in luminal epithelial cells on day 4. The protein of Ang-2 was mainly expressed in the cytoplasma of glandular epithelia and stromal cells. With gestation time, the positive reactions of Ang-1/-2 were stronger in the endometria of the pregnant mice （P〈0.01）. In situ hybridization showed Ang-I mRNA in stromal cells on day 2. Hybridization signal was localized in both stromal cells and vessel epithelial cells on day 4; Ang-2 mRNA was expressed in stromal cells and glandular epithelia on day 2; high mRNA levels appeared in stromal cells, glandular epithelia and vascular endothelia on day 4; an increasing in mRNA expression of Ang-1/-2 was observed on day 6 and day 8 （P〈0.01）. It is suggested that Ang-1/-2 may play an important role in the cross-talk between blastocyst and maternal endometrium during the process of embryo implantation.
基金partially supported by a grant from the National"Ten Times Five Years"Key Technologies Research Development Program of China(No.2004BA720A33-01)
文摘Brain-derived neurotrophic factor (BDNF) can promote developmental competence in mammalian oocytes during in vitro maturation (IVM),but the role of BDNF in oocyte maturation at cellular level is not still clear.In this study,mouse cumulus-enclosed oocytes subjected to IVM were fertilized and cultured to blastocyst stage.Meiotic spindle configuration and cortical granules distribution during oocyte maturation in vitro were assessed by using immunofluorescence and laser confocal microscopy.The results showed that BDNF contributed to the complete preimplantation development of mouse oocytes compared to the control oocytes (13.78% vs.5.92%;P【0.05).Further,BDNF did not accelerate nuclear maturation of IVM oocytes.For the BDNF-treated oocytes at meiosis Ⅰ,Meiotic spindle areas were significantly smaller and the number of cytoplasmic microtubule organizing centers was greater than that in the control,and the percentages of oocytes showed spindles positioned near the oolemma and a well-formed cortical granule-free domain were significantly higher than that of the control.These morphological characteristics of the BDNF-treated oocytes were much closer to the oocytes matured in vivo than those of the control oocytes.In conclusion,BDNF can promote the developmental competence of mouse IVM oocytes,by improving the meiotic spindle configuration and location and cortical granules distribution at meiosis Ⅰ.
基金This work is supported by a grant of the National "Tenth Five Years" Key Technologies R&D Programme,China(No.2004BA720A33-01).
文摘Objective To explore the role of urokinase-type plasminogen activator（uPA） in precontact sperm-egg communication and fertility of mice in vitro. Methods Firstly, sperm chemotaxis （SC） induced by uPA was assayed by measuring the sperm densities in capillaries with a descending gradient or no gradient of uPA respectively. Secondly, the role of uPAR that exists in sperm plasma membrane in SC was studied by examining the change of sperm density in capillary after incubating spermatozoa with anti-uPAR antibody. Thirdly, SC induced by eggs, which had been treated with uPA, PAl-1 and anti-uPAR beforehand respectively, was assayed to study the role of uPA in PSEC. Lastly, the fertilization capability of spermatozoa treated with uPA was examined by counting the number of fertilized eggs. Results 1）The density of spermatozoa that migrated down the gradient of uPA into the capillary was significantly lower than that into the capillary containing no-gradient uPA. 2） When uPAR of spermatozoa was inhibited by anti-uPAR antibody, the density of spermatozoa that migrated into the capillary with ascending gradient of uPA decreased correspondingly. 3） The density of spermatozoa attracted by eggs, which were treated with uPA beforehand, increased significantly than that of attracted by non-treated eggs. On the contrary, the sperm density decreased correspondingly when the egg was treated with PAI-1. 4） The number of fertilized eggs increased significantly after the spermatozoa used here was treated with uPA beforehand. Conclusion uPA could induce SC of mice sperm in vitro through the uPAR on its membrane, enhance the capability of egg inducing SC, and promote spermatozoa to fertilize eggs. Thus, uPA may act as an attractant in PSEC, increase the chance encounter of spermatozoa and eggs, therefore, enhance the fertility success correspondingly. This study, in some degree, provides an evidence that uPA may be used as a new medicine and diagnostic reagent for male infertility.
基金a grant from National Natu-ral Sciences Foundation of China (No. 30371829)
文摘In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor （LIF） in mice with embryonic implantation dysfunction （EID）, 120 Kunming mice post coition were randomized into three groups： normal control group, model group and traditional Chinese medicine group （TCM group） （n=40 in each group）. Uterus was collected on the pregnancy day （Pd） 4, 5, 6 after an intravenous injection of Evan＇s blue. The endometrium was dyed by Evan＇s blue and the mean points of response were observed on Pd 5. The expression of LIF mRNA and protein was detected by RT-PCR and immunohistochemistry respectively and analyzed statistically by image system. The results showed that the number of implantation sites in model group was remarkably less than in normal control group and TCM group. There was no significant difference between normal control group and TCM group. The expression of LIF mRNA and protein in model group was delayed. Bushenantai recipe could increase the expression of LIF mRNA and protein in endometria of mice with EID. It was suggested that Bushenantai recipe could improve embryo implantation of mice with EID by promoting the endometrial LIF expression and endometrial decidualization.
基金supported by National Key Research&Development Program(Grant No.2016YFC1000200,No.2016YFC1000204,and No.2018YFC1004200)the State Key Program of National Natural Science of China(Grant No.31530047)+2 种基金National Natural Science Foundation of China(Grant No.81602927)Innovation Fund of State Key Laboratory of Reproductive Medicine(Grant No.SKLRMGC201802)Top-notch Academic Programs Project of Jiangsu Higher Education Institutions(Grant No.PPZY2015A067)。
文摘There were few studies of cumulative live birth rates(CLBRs) based on multicenter reproductive clinical data from the general Chinese population.Here we report a retrospective cohort study,including 14 311 women with17 315 cycles,in three reproductive centers to evaluate two estimated parameters of CLBRs with multiple transfer cycles of in vitro fertilization(IVF)/intracytoplasmic sperm injection(ICSI) in a Chinese population.We found that CLBRs were related to female age and endometrial thickness.By the fourth transfer cycle,the conservative and optimal estimates of CLBRs were 52.95% and 77.30% in women under 30 years of age,and 18.17% and26.51% in those 37 years of age or older,respectively.The two estimates were 44.70% and 63.15% in women with endometrial thickness more than 7 mm,and 32.05% and 46.18% in those with less than 7 mm,respectively.In addition,body mass index(BMI),duration of infertility,and infertility diagnoses may also be related to CLBRs on certain conditions.The findings from this study on CLBRs after multiple transfer cycles of IVF/ICSI treatment on different conditions in the Chinese population should be beneficial to both infertile couples and clinicians.
