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Abstract Background Endometrial cavity fluid (ECF) is a fluid accumulation within the endometrial cavity. The significance of ECF remains unclear during the program of in vitro fertilization-embryo transfer (IVF-ET). The aim of the present study was to investigate the associated factors to ECF, visualized through ultrasound at the day of oocyte retrieval, and the relevant impact on the outcome of IVF-ET. Methods From the clinical data of 1557 infertility patients for IVF-ET program, 46 ECF patients were retrospectively selected as the ECF group; and another 134 patients with a bilateral salpingectomy and without ECF, selected as the control group. The demographics and the outcome of IVF-ET were compared between the two groups. Results The incidence of ECF was 2.95% (46/1557). Over half (28/46, 60.87%) of ECF patients had tubal infertility. Only 12 Of 46 ECF patients (26.09%) had visible hydrosalpinx on ultrasonography before ovarian stimulation. The cycle cancellation rate (4/46, 8.69%) of ECF group was not significantly higher than that of the control group (6/134, 4.48%; P > 0.05). Reasons for cycle cancellation in both groups were all the high risk of ovarian hyperstimulation syndrome (OHSS). No significant difference was found in clinical pregnancy rate between the patients with their ECF 0.05). No clinical pregnancy was found among those patients with their ECF equal or higher 3.5 mm in APD. Conclusions It was tubal infertility, not hydrosalpinx, which was related to the development of ECF. Excessive ECF (equal or higher 3.5 mm in APD) at the day of oocyte retrieval would have a negative impact on the outcome of IVF-ET.

A simple fluorescence biosensor is developed based on the enzyme-assisted cascade amplification strategy. The amplification system consists of a hairpin-structure DNA (H-DNA) and exonuclease III. The target DNA can hybridize with the H-DNA and initiate exonuclease III-assisted target recycling amplification to generate abundant G-rich DNA (G-DNA). One region of G-DNA is designed to possess the same sequence as target DNA. Thus, the G-DNA can also hybridize with H-DNA and initiate the digestion of H-DNA. The cascade strategy in this amplification system causes the concentration of G-DNA to grow exponentially. The fluorescence intensity of N-methylmesoporphyrin IX (NMM) is highly enhanced due to the formation of G-quadruplex configuration. Under optimal conditions, the cascade system could achieve an admirable sensitivity with a detection limit of 52 fM for HIV DNA, and guarantees a satisfactory specificity. Moreover, the cascade system could be implemented for other target DNA detections by substituting the recognition region of the H-DNA. In this way, a detection limit of 65 fM for HBV DNA could be achieved by the cascade system. The target DNA analysis in a real serum sample further indicates that this biosensor has potential for future application in clinical diagnosis. [ABSTRACT FROM AUTHOR]

Abstract: Silk fibroin is a kind of polypeptide with functional amino acids in its structure. The electric charges in its molecular chains originating from the dissociation of acidic groups, i.e., hydroxyl, phenol and carboxyl, provide vast potentials for the retention of metal species of interest. In this study, the selective retention of Cu2+ with silk fibroin at pH 6.0 was investigated and a novel on-line procedure for separation/preconcentration of Cu2+ from complex sample matrices was thus developed by using a sequential injection system with an electrothermal atomic absorption spectrometry. A novel concept of enrichment index (EI), i.e., defined as enrichment factor (EF) obtained by consuming unity of sample volume (ml), was proposed for evaluating the enrichment efficiency of a flow-based preconcentration procedure. With a sampling volume of 900μl, an EI of 30.3 (EF=27.3) was achieved, which was much improved as compared to that of reported procedures. A detection limit of 8.0ngl−1 was achieved within a linear range of 0.025–1.5μgl−1 along with a precision of 2.2% R.S.D. at 0.5μgl−1. The practical applicability of this procedure was validated by analyzing a certified reference material of riverine water (GBW08608) and a certified reference material of seawater (NASS-5) achieving satisfactory agreements between the certified and the obtained values. A spiking recovery was also performed by using a cave water sample. [Copyright &y& Elsevier]

Abstract: The dominant role played by flow injection/sequential injection (FI/SI, including lab-on-valve, LOV) in automatic on-line sample pretreatments coupling to various detection techniques is amply demonstrated by the large number of publications it has given rise to. Among these, its hyphenation with hydride/vapor generation atomic fluorescence spectrometry (HG/VG-AFS) has become one of the most attractive sub-branches during the last years, attributed not only to the high sensitivity of this technique, but also to the superb separation capability of hydride/vapor forming elements from complex sample matrices. In addition, it also provides potentials for the speciation of the elements of interest. It is worth mentioning that quite a few novel developments of sample pretreatment have emerged recently, which attracted extensive attentions from the related fields of research. The aim of this mini-review is thus to illustrate the state-of-the-art progress of implementing flow injection/sequential injection and miniaturized lab-on-valve systems for on-line hydride/vapor generation separation and preconcentration of vapor forming elements followed with detection by atomic fluorescence spectrometry, within the period from 2004 up to now. Future perspectives in this field are also discussed. [Copyright &y& Elsevier]

Objective: To evaluate the levels of bone morphogenetic protein-15 (BMP-15) in human follicular fluid (FF) and its association with response to ovarian stimulation., Methods: Western blotting was performed to determine the levels of BMP-15 in FF obtained from follicle aspirates in 70 patients undergoing IVF treatment. According to the response to ovarian stimulation the patients were divided into poor responder group and normal responder group., Result: BMP-15 levels in FF of poor responders were significantly higher than those in normal responders (1.01 +/- 0.34 vs 0.77 +/- 0.24, P<0.01)., Conclusion: Increased levels of BMP-15 in FF may be associated with poor response to ovarian stimulation.

Objective: To investigate the possible roles of cyclin B1 and cyclin-dependent kanase1 cyclin-dependent kanase1 play in the pathogenesis and development of endometriosis and their association with the ovarian hormones., Methods: Twenty-nine specimens of ectopic endometrium tissues and 20 specimens of eutopic endometrium tissues were obtained from 29 patients with endometriosis, aged 24 approximately 46. Thirty specimens of endometrium form 30 women without endometriosis were used as controls. The intracellular location of cyclin B1 and Cdc2 was detected by microscopy. Western blotting, RT-PCR, and immunohistochemistry were employed to examine the mRNA expression and protein expression of cyclin B1 and Cdc2. Serum estrogen and progestogen were detected., Results: The expression level of cyclin B1 in the ectopic endometrium from the women with endometriosis was significantly higher than that in the ectopic endometrium tissues from the women with and without endometriosis (both P < 0.05). No significant difference was found between the expression level of cyclin B1 in the ectopic endometrium tissues from the women with and without endometriosis (both P > 0.05). The Cdc2 expression levels were not significantly different among th3 3 groups and the proliferative and secretary stage of endometrium (all P > 0.05). Cyclin B1 expression level was positively correlated with the serum estrogen level, and negatively correlated with the serum progestogen level, and Cdc2 expression was not correlated with the serum sex hormone levels., Conclusion: The expression of cyclin B1 in the ectopic endometrium is higher than normal. Cyclin B1 may be involved in the proliferation of endometrium in endometriosis.

Objective: To investigate whether the limited ovarian stimulation (LOS) could avoid the severe ovarian hyperstimulation syndrome (OHSS) in patients with high risk of OHSS., Methods: Ten infertile patients were diagnosed as polycystic ovarian syndrome (PCOS) with high risk of OHSS. All of them were arranged LOS cycles, a long-induction protocol using gonadotropin releasing hormone agonist (GnRHa) for controlled ovarian hyperstimulation. A full dose of human chorionic gonadotropin (10,000 U) was administered when the leading follicle reached a mean diameter of 12.0 - 14.5 mm. Oocytes were retrieved 36 hours later, followed by conventional in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) and embryo transfer., Results: Ten patients produced 11 - 35 oocytes, including 4 - 28 oocytes of metaphase II. Embryos were transferred in all of ten patients and eight clinical pregnancies were diagnosed. Unfortunately, ectopic pregnancy occurred in one of the patients. None of them experienced the symptoms of OHSS., Conclusions: LOS may play a valuable role in preventing OHSS and achieving enough mature oocytes for the patients of PCOS. Furthermore, this protocol would not reduce the pregnancy rate of the patients.

Objective: To examine the expression of aquaporin 2 (AQP2) in human endometrium., Methods: Specimens of human endometrium were collected from 87 women at different menstrual cycles, aged 30 +/- 3, 23 cases in the proliferative phase, 30 cases in the early secretory phase and 34 cases in the mid-secretory phase respectively. Immunohistochemistry and Western blotting were utilized to detect the localization and expression of AQP2 in the endometrium. Reverse transcription-polymerase chain reaction (RT-PCR) was employed to detect its messenger RNA. The PCR product was cloned and sequenced., Results: Positive immunoreactivity of AQP2 was found in the epithelia cells and glandular epithelial cells of all specimens of human endometrium at different phases of the menstrual cycle, and all stromal cells were not stained. The reactive substance was primarily distributed in the membrane and cytoplasm, but not in the nuclei of all positive cells. In Western blotting showed dominant bands with relative molecular weight between 35,000 Da and 50,000 Da, which was corresponded to the glycosylated form of AQP2 by the positive control from rat kidney. Semi-quantitative analysis showed that the relative expression of AQP2 in the mid-secretory phase was 1.63 +/- 0.15, significantly higher than those in the early secretory phase (1.33 +/- 0.14, P < 0.05) and that in the proliferative phase (1.03 +/- 0.10, P < 0.01). Message RNA was found out in all cases by RT-PCR and the PCR product was confirmed in nearly exact (99%) consistency with the GenBank by sequencing. AQP2 mRNA was expressed in all normal endometrium at different phases, and was weakly expressed in the endometrium at the proliferative phase. Sequencing showed that the AQP2 sequence was 99% homologous with that in the GenBank., Conclusion: AQP2 expression in the human endometrium suggests that AQP2 may be involved in the regulation of uterine fluid homeostasis influenced by ovarian steroid hormones in the menstrual cycle.

Objective: To investigate the expressions of peripheral lymph node addressin(PNAd) and GlcNAc-6-sulfotransferase (GlcNAc6ST) in endometrium and their impacts on implantation., Methods: PNAd expression in endometrium was examined by immunohistochemistry and Western Blot from 75 women (12 from healthy women, in proliferative phase; 63 from sterile women, of whom, 27 were in early-secretory and 36 in mid-secretory phase). GlcNAc6ST mRNA was examined by real-time PCR in 41 sterile women. The 63 sterile women had underwent IVF-ET and were consequently divided into clinical pregnant (29 cases) and nonpregnant (34 cases) groups., Results: (1) PNAd localized to the membrane and cytoplasm of luminal and glandular epithelia. Staining was patchy and much less intense during the proliferative phase than during the secretory phase. In Western Blot of PNAd, four bands appeared, which were Sgp200, CD34, MAdCAM-1, GlyCAM-1 respectively, and each was positively correlated with the others significantly. The former three molecular levels were significantly higher during the secretory phase as compared with the proliferative phase. Message RNA of GlcNAc6ST was positive in all cases and showed no correlation with any component of PNAd. (2)The expressions of CD34 and GlyCAM-1, but not Sgp200 and MAdCAM-1, were significantly higher in pregnant women than in nonpregnant ones. However, the GlcNAc6ST mRNA level did not differ between groups. (3) No significant difference was found in female age, methods of fertilization, thickness of endometrium on day hCG, cumulative embryo score(CES) and mean score of transferred embryo(MSTE) between the groups., Conclusion: PNAd expression in the human endometrium fluctuates with the menstrual cycle. Elevated CD34 and GlyCAM-1 during the secretory phase might be stimulative factors for embryo implantation. Defect in PNAd expression may account for a portion of unexplained infertility.