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Qin Y.-N.,Jiangxi Provincial Maternal and Child Health Hospital | He D.-M.,Jiangxi Provincial Maternal and Child Health Hospital | Zhang Z.-Y.,Jiangxi Provincial Maternal and Child Health Hospital | Yu X.-H.,Jiangxi Provincial Maternal and Child Health Hospital
International Journal of Clinical and Experimental Pathology | Year: 2017

Cervical cancer is one of the most common gynecological carcinoma, which seriously threaten the life and health of women globally. Previous studies have shown that β-catenin abnormalities in the expression and localization is closely related to the the pathogenesis and development of cervical cancer. In the Wnt/β-catenin signaling pathway, casein kinase 1 (CK1) protein family had both inhibitory and activated functions. As one of the seven isoforms of the CK1 family, CK1δ function is poorly defined. Here, by using tissue microarray, we found that, compared with control (chronic cervicitis tissue), CK1δ protein expression level was significantly elevated in 88 cervical squamous cell carcinoma (CSCC) tissues (7.7% vs. 58.0%, P < 0.001). The increased CK1δ expression was associated with deep cervical stromal invasion in patients with cervical cancer (P=0.015). Besides, the expression levels of CK1δ and β-catenin in cervical cancer tissues was positively correlated in CSCC tissues (P < 0.001). Therefore, we hypothesized that CK1δ overexpression may contribute to cervical cancer progression through activating the Wnt/β-catenin signaling pathway. Based on above experimental results, we can get a deeper understanding of the role of CK1δ in the Wnt/β-catenin signaling pathway and cervical cancer, and to find new biomarkers and therapeutic targets for the diagnosis of cervical cancer.

PubMed | Shanghai JiaoTong University, Shandong University, Nanjing Medical University, Anhui Medical University and 5 more.
Type: | Journal: Human reproduction (Oxford, England) | Year: 2016

Do oral contraceptives (OCs) and progestins impact live birth rate ofIVF when used for cycle scheduling in women with polycystic ovary syndrome (PCOS)?OCs used for scheduling IVF cycle were associated with lowered rates of pregnancy and live birth after fresh embryo transfer, whereas progestins used for this purpose yield higher rates of pregnancy and live birth than OCs.Due to oligo-menorrhea in PCOS, OCs and progestin are extensively used to schedule the start of an IVF cycle in women with PCOS. Little is known about the effect of such pretreatments on outcomes, especially, the rate of live birth.This was a nested cohort study and secondary analysis of a multicenter randomized trial, which was designed to compare live birth rate after fresh embryo transfer vs frozen embryo transfer (FET) in women with PCOS (Frefro-PCOS). A total of 1508 women were enrolled from 14 centers between June 2013 and May 2014.At the discretion of local investigators, subjects were instructed to wait for spontaneous menses (Control group, n = 323), or were prescribed progestins (P group, n = 283) or OCs (OCs group, n = 902) to induce menstruation prior to the start of ovarian stimulation. GnRH antagonist protocol was initiated at Day 2 or 3 of induced or spontaneous menses cycle. The rates of pregnancy, pregnancy loss and live birth after either fresh embryo transfer or FET were compared among these three groups.With fresh embryo transfer, women with OC-induced menses had lower rates of clinical pregnancy (48.8% vs 63.6%, relative rate (RR): 0.77, 95% CI: 0.66-0.89) and live birth (36.1% vs 48.1%, RR: 0.75, 95% CI: 0.61-0.92) than women with spontaneous menses. With freeze-all and deferred FET, women with OC-induced menses had a similar pregnancy rate but a higher pregnancy loss rate (27.7% vs 13.0%, RR: 2.13, 95% CI: 1.28-3.52) after FET than women with spontaneous menses. The live birth rate after FET in women with OC-induced menses, progestin-induced menses and spontaneous menses was 49.4%, 50.7% and 60.2%, respectively (P = 0.06). Progestin-induced menses was associated with similar rates of pregnancy, pregnancy loss and live birth after transfer of either fresh or frozen embryos compared with spontaneous menses. Multivariate logistic regression analysis showed that OCs used for menses induction was associated with lower rate of live birth.The methods for menses induction were not assigned randomly, thus selection bias was highly likely because of the study design and significant differences that were observed in the baseline characteristics of the women in the different groups. The mean BMI in this study population was relatively normal; the applicability of this result to obese PCOS women needs to be evaluated in further study.Our results suggest that either waiting for a spontaneous menses or using progestin is a better option than using OCs to induce menses in women with PCOS prior to ovarian stimulation using GnRH antagonist protocol for IVF. Further randomized controlled studies are needed to confirm our findings.This study was funded by National Basic Research Program of China (973 Program) (2012CB944700), the State Key Program of National Natural Science Foundation of China (81430029), National Natural Science Foundation of China (81471428) and Thousand Talents Program (Drs Legro and Zhang H). Dr Legro reports receiving consulting fees from Euroscreen, Kindex, Bayer and Millendo Pharmaceuticals and research funding from Ferring. Others report no disclosures.Frefro-PCOS was registered at Clinicaltrials.gov: NCT01841528.

PubMed | Nanchang University, Jiangxi Childrens Hospital, Gannan Medical University and Jiangxi Provincial Maternal and Child Health Hospital
Type: | Journal: Ultrasound in medicine & biology | Year: 2016

The aim of this study was to investigate the clinical efficacy of high-intensity focused ultrasound (HIFU) for the treatment of a cesarean scar pregnancy compared with uterine artery embolization (UAE) and intra-arterial methotrexate infusion combined with uterine curettage. In this retrospective cohort study, 31 patients were treated with HIFU (HIFU group), and 45 patients were treated with UAE (UAE group). We compared the treatment and recovery of the patients, including follow-up. After UAE treatment, serum levels of the subunit of human chorionic gonadotropin declined significantly on the first day, and the residual lesions disappeared in 3-17wk. One patient underwent hysterectomy; intrauterine adhesions were found by hysteroscopic examination after 6mo in 2 patients, whose menstrual function did not return to normal. The remainder of the 42 patients recovered normal menstrual functioning during the 3- to 18-wk follow-up. In the patients who underwent HIFU treatment, serum -HCG levels did not decline rapidly; serum -HCG levels increased in many patients and then declined to normal steadily within 2-12wk. Lesions detached in 3-14wk in all patients, and menstrual functioning was recovered in 3-9wk without uterine curettage. Compared with the UAE group, the HIFU group had less pain and fewer complications; the patients in the HIFU group were not hospitalized or anesthetized and had lower costs. HIFU is an efficient, tolerable and non-invasive treatment.

PubMed | Jiangxi Provincial Maternal and Child Health Hospital, Shanghai Tobacco Group Corporation of CNTC and CAS Dalian Institute of Chemical Physics
Type: | Journal: Chemico-biological interactions | Year: 2016

N-Nitrosonornicotine (NNN) is considered to be one of the most carcinogenic compounds of the four conventionally measured tobacco-specific N-nitrosamines (TSNAs). In order to evaluate the significance of metabolic activation for the carcinogenic potential of NNN, its catalysis by different phase I enzymes and its interaction with nicotine and nicotine-derived TSNAs need to be investigated. Using an invitro model system, NNN was found to interact with various cytochrome P450 enzymes, predominantly CYP2A13. Mass-spectrometric analysis confirmed the presence of various predicted NNN metabolites, including 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB) and 4-hydroxy-4-(3-pyridyl)-butyric acid (hydroxy acid) but little amount of 4-oxo-4-(3-pyridyl) butanal (OPB), which was somewhat different from invitro NNK metabolism. Addition of nicotine, N-Nitrosoanatabine (NAT), N-Nitrosoanabasine (NAB) resulted in a competitive inhibition for NNN metabolism. The inhibition constant Ki value was calculated as 0.98M (nicotine), 1.37M (NAT), 0.71M (NAB) for HPB formation, 1.35M (nicotine), 1.35M (NAT), 1.01M (NAB) for hydroxy acid formation and 8.40M (nicotine), 3.40M (NAT), 3.04M (NAB) for OPB formation, respectively. These results implied that CYP2A13 is the most efficient enzyme to metabolize NNN invitro and structurally similar tobacco constitutes including nicotine, NAT and NAB influence the metabolic activation of NNN, which may further interfere in its carcinogenicity.

PubMed | Jiangxi Provincial Maternal and Child Health Hospital and Nanchang University
Type: | Journal: Gene | Year: 2016

Adenomyosis is a common benign gynecological condition in female reproductive tract and the detailed molecular etiology remains largely elusive. Previous studies implicated that deregulated expression of DNA (cytosine-5)-methyltransferase 3A (DNMT3A), a de novo DNA methyltransferase, might be involved in the pathogenesis of adenomyosis. Meanwhile, ectopic endometrial stromal cells (EESCs) were suggested to play crucial roles in adenomyosis. Herein, we evaluated the expression of DNMT3A protein in 36 ectopic endometriums with adenomyosis and 37 eutopic endometriums in controls with Western blotting (WB) or immunohistochemistry (IHC), we found that the expression of DNMT3A was significantly decreased in the ectopic endometriums and EESCs in adenomyosis relative to that of eutopic endometriums and EESCs in control samples, respectively. In addition, our functional assays revealed that overexpression of DNMT3A suppressed cell proliferation and invasion, while knockdown of DNMT3A enhanced cell proliferation and invasion in EESCs. Taken together, our results suggested that DNMT3A expression was decreased in ectopic endometriums and EESCs in adenomyosis, and we provided the first evidence that decreased DNMT3A expression in EESCs facilitated the development of adenomyosis via enhanced cell growth and invasion.

PubMed | North University of China, Central South University, Liaoning Medical University, Nanchang University and 2 more.
Type: Journal Article | Journal: Molecular neurobiology | Year: 2016

In this study, we investigated the influence of elevated RAS expression on the growth of meningioma in vivo and in vitro. The IOMM-LEE cells, representing a cell line derived from malignant meningioma, were divided into blank control group (cells without any drug treatment), negative control group (cells treated with an equal volume of normal saline to replace drug), and farnesyl thiosalicylic acid (FTS)-treated group (cells treated with FTS). Methyl-thiazole-tetrazolium bromide (MTT) assay and flow cytometer (with cells after FTS (75mol/L) treatment for 48h) were utilized to determine the proliferation and apoptosis, respectively, of IOMM-LEE cells after RAS inhibition. Western blot analysis was used for semi-quantitative analysis of p-ERK and p-AKT levels. Animal model of human meningioma was established with sub-renal capsule transplantation, and mice were divided into two groups: experimental group (50mg/kg group, 75mg/kg group, and 100 mg/kg, hypodermic injection with FTS) and control group. Proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry (IHC). Western blot analysis was used for detecting ERK and AKT signal pathway. The proliferation of IOMM-LEE cells decreased dramatically and apoptosis rate increased significantly in FTS-treated group compared to blank control group and negative control group (all P<0.05). At FTS concentration of 75mol/L, the apoptosis rate of IOMM-LEE cells reduced significantly over time (P<0.05). Cell cycle analysis showed that IOMM-LEE cells exhibited G1-arrest in the FTS-treated group, compared to no cell-cycle arrest in blank control group and the negative control group (P<0.05). Further, significantly decreased ERK and AKT phosphorylation levels were detected in IOMM-Lee cells after FTS (75mol/L) treatment for 48h, compared to blank control group and negative control group (P<0.05). The results in vivo experiments showed that after FTS treatment, tumor volume, PCNA LI, and the levels of p-ERK and p-Akt decreased significantly in 75mg/kg group and 100mg/kg group when compared with the control group and 50mg/kg group (all P<0.05). Our findings provide strong evidence that RAS protein is highly expressed in meningioma cells, and the RAS activity is inhibited by downregulating ERK and AKT signal pathway, which may further inhibit the growth of meningioma.

PubMed | Jiangxi Provincial Maternal and Child Health Hospital
Type: Journal Article | Journal: European review for medical and pharmacological sciences | Year: 2017

In humans, stem cell factor (SCF), produced by cumulus granulosa cells during the follicular phase, plays a crucial role in follicular development. Remarkably, polycystic ovary syndrome (PCOS), one of the main reasons affecting women fertility, is accompanied by some abnormal follicles. Is there a relationship between SCF and PCOS? This study aimed to compare the expression of SCF in follicle and serum from patients with and without PCOS undergoing in vitro fertilization (IVF) treatment and to investigate the potential relationship between aberrant SCF expression and PCOS.Serum, follicular fluid (FF) samples and granulosa cells (GCs) from 48 patients with PCOS (PCOS group) and 62 normal ovulatory patients (control group) were collected. SCF was evaluated in FF, serum, and GCs by using enzyme-linked immunosorbent assay, immunofluorescence staining, Western blot and real-time PCR. The rates of metaphase II (MII) oocyte, fertilization, embryo cleavage and high-quality embryo between PCOS group and control group were also analyzed.The rates of MII oocyte and fertilization were significantly lower in PCOS group than those in control group (p < 0.05). No difference was observed for the rate of embryo cleavage and high-quality embryo in these two groups. The concentrations of SCF in serum and FF from PCOS patients were remarkably lower than those in the controls (p < 0.05). Moreover, the expressions of SCF protein and SCF mRNA in GCs from PCOS patients were also decreased compared with the controls (p < 0.05).PCOS patients showed a reduced SCF expression in serum and follicle, which might be associated with oocyte dysmaturity and low fertilization rate.

PubMed | Nanchang University, Jiangxi Childrens Hospital and Jiangxi Provincial Maternal and Child Health Hospital
Type: | Journal: International immunopharmacology | Year: 2016

The precision-cut kidney slice (PCKS) model appears to be a useful ex vivo model of renal fibrosis. However, little in-depth molecular investigation on the PCKS model has been performed. Therefore, the aim of this study will be to investigate and validate the molecular validity of this model.The PCKS model was constructed in male C57BL/6 mice. To induce renal fibrosis, PCKS were incubated in recombinant human TGF-1 for 48 h. Protein expression of phosphorylated Smad2 (p-Smad2, cytosolic and nuclear), Smad7, phosphorylated ERK1 (p-ERK1), phosphorylated ERK2 (p-ERK2), and phosphorylated p38 MAPK (p-p38 MAPK) was measured using Western blotting. To assess Smad2/3 heteromeric complex formation and phosphorylated Smad3 (p-Smad3) expression, immunoprecipitation was performed with an anti-Smad2 or an anti-Smad3 antibody, respectively, prior to Western blotting.p-Smad2 and p-Smad3 were significantly upregulated in the PCKS model relative to control (p<0.05). However, we found no significant difference in Smad7 expression between the PCKS model and control (p>0.05). The PCKS model demonstrated significantly greater Smad2/3 complex formation and nuclear translocation relative to control (p<0.05). The PCKS model showed significantly greater expression of p-ERK1, p-ERK2, and p-p38 MAPK relative to control (p<0.05).The PCKS model displays several well-established molecular markers of renal fibrosis. However, the PCKS model failed to display Smad7 downregulation and appears to display over-activation of p-Smad2 and p-Smad3 as well as under-activation of ERK1/2 and p38 MAPK signaling vis--vis the well-established in vivo unilateral ureteric obstruction model of renal fibrosis.

PubMed | Nanchang University, Jiangxi Province Tumor Hospital and Jiangxi Provincial Maternal and Child Health Hospital
Type: Journal Article | Journal: Biochemical and biophysical research communications | Year: 2016

Ketamine, a dissociative anesthetic, which was widely used in human and animal medicine, has become a popular recreational drug, as it can induce hallucinatory effects. Ketamine abuse can cause serious damage to many aspects of the organism, mainly reflected in the nervous system and urinary system. It has also been reported that ketamine can impair the male genital system. However, the detailed effect of ketamine on human spermatozoa remains unclear. Thus, we investigated the invitro effects of ketamine on human sperm functions, to elucidate the underlying mechanism. Human sperm were treated invitro with different concentrations of ketamine (0, 0.125, 0.25, 0.5, 1g/L). The results showed that 0.25-1g/L ketamine inhibited sperm total motility, progressive motility and linear velocity, in a dose-dependent manner. In addition, the sperms ability to penetrate viscous medium and the progesterone-induced acrosome reaction were significantly inhibited by ketamine. Ketamine did not affect sperm viability, capacitation and spontaneous acrosome reaction. The intracellular calcium concentration ([Ca(2+)]i), which is a central factor in the regulation of human sperm function, was decreased by ketamine (0.125-1g/L) in a dose-dependent manner. Furthermore, the currents of the sperm-specific Ca(2+) channel, CatSper, which modulates Ca(2+) influx in sperm, were inhibited by ketamine (0.125-1g/L) in a dose-dependent manner. Our findings suggest that ketamine induces its toxic effects on human sperm functions by reducing sperm [Ca(2+)]i through inhibition of CatSper channel.

PubMed | Jiangxi Provincial Maternal and Child Health Hospital
Type: Journal Article | Journal: Journal of mass spectrometry : JMS | Year: 2016

Newborn screening is one of public health concerns designed to screen infants shortly after birth. Prenatal exposure to tobacco smoke such as nicotine has been reported to affect babies. Levels of nicotine and cotinine in meconium were widely used to evaluate the tobacco exposure of foetuses during pregnancy in a polluted environment. In this study, medical swabs were applied by using touch spray-mass spectrometry (TS-MS) to collect meconium from newborn infants for detection of nicotine and cotinine. Parameters such as choice of spray solvents, solvent volume and collision energy for screening of nicotine and cotinine were optimized. The limits of detection, reproducibility and matrix effect for analysis of meconium were also investigated. In this study, the levels of nicotine and cotinine in 54 puerpera volunteers were screened by TS-MS and were validated by using traditional liquid chromatography-mass spectrometry. These results showed that medical swab TS-MS would be useful for newborn screening of nicotine and cotinine in meconium with high reproducibility, speed, sensitivity and specificity. The use of disposable medical swabs involves no sample preparation and no chromatographic separation, significantly reducing the cost and time required for screening a large number of clinical sample. Copyright 2016 John Wiley & Sons, Ltd.

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