Using the stopped-flow circular dichroism (SFCD) technique, we investigate the kinetics of the pH-induced folding and unfolding process of the DNA i-motif. The results show that the molecule can fold or unfold on a time scale of 100 ms when the solution pH is changed. It is also found that the folding and unfolding rates strongly depend on the solution pH. On the basis of quantitative data, we propose theoretical models to decipher the folding and unfolding kinetics. Our models suggest that the cooperativity of protons is crucial for both the folding and unfolding process. In the unfolding process, the cooperative neutralization of two protons (out of the total six protons in the i-motif molecule) is the only rate-limiting step. In the folding process, there exists a critical step in which three protons bind cooperatively to the DNA strand. These results offer an in-depth understanding of the folding and unfolding kinetics of the DNA i-motif and may give precise guidance for constructing novel nanodevices based on the DNA i-motif.

Recent evidence shows that the outcome of rescue intracytoplasmic sperm injection (ICSI) is unsatisfactory on account of a poor clinical pregnancy rate. These outcomes may be due to either the in-vitro ageing of cultured oocytes before ICSI or the asynchrony between the embryo developmental stage and the endometrial secretory pattern. To address the latter issue, this study performed a retrospective analysis of 534 fresh cycles after rescue ICSI and 64 frozen-thawed cycles in subsequent treatment. Rescue ICSI cycles were divided into three groups: group I included 469 fresh embryo-transfer (FET) cycles; group II included 74 FET cycles in which supernumerary good-quality embryos were also cryopreserved; and group III included 64 frozen-thawed transfer cycles. Group III was considered to have achieved better synchronization than group II. As a result, significantly higher clinical pregnancy (29.69%, 19/64 versus 10.81%, 8/74) and implantation (13.33%, 22/165 versus 5.13%, 8/156) rates were achieved in group III compared with group II (both P<0.05). Therefore, synchronization of embryo development with the endometrium is considered a contributing factor for rescue ICSI outcome. It is recommended that embryos derived from rescue ICSI cycles should be cryopreserved and subsequently used in frozen-thawed cycles. Intracytoplasmic sperm injection (ICSI) of unfertilized 1-day-old oocytes, called rescue ICSI, has frequently been performed in some infertility centres, when fertilization failure sometimes occurs in conventional IVF cycles. Recent studies showed that the outcome of rescue ICSI was unsatisfactory due to poor clinical pregnancy rates. One reason could be asynchrony between the embryo developmental stage and the endometrial secretory pattern. To address this issue, we performed a retrospective analysis of 534 fresh cycles after rescue ICSI (from January 2006 to January 2011) and 64 frozen-thawed transfer cycles in subsequent treatment (from January 2006 to May 2011) in our infertility centre. In this study, rescue ICSI cycles were divided into three groups. As there was no significant difference in women's age (31.22 ± 3.38 versus 31.11 ± 3.27 years) between groups II and III, we principally compared these two groups. Group II included 74 fresh embryo transfer cycles, in which supernumerary good-quality embryos were cryopreserved, and group III included 64 frozen-thawed transfer cycles. Group III was considered to have better synchronization than group II. As a result, significantly higher clinical pregnancy (29.69% versus 10.81%) and implantation (13.33% versus 5.13%) rates were achieved in group III compared with group II. Therefore, endometrial synchronization is considered a contributing factor for rescue ICSI outcome and embryos derived from rescue ICSI cycles should be cryopreserved and subsequently used in frozen-thawed cycles.

Transfer of 0PN embryos from fresh or frozen-thawed cycles results in pregnancies and live births. Nonpronuclear embryos have a lower IR than 2PN embryos, but if the embryos are cultured to the blastocyst stage and then are frozen, their IRs approach that of 2PN embryos in subsequent frozen-thawed cycles. The culture of 0PN embryos to the blastocyst stage may select for embryos with a near-normal IR.

High incidence of polyspermy is still a major problem in the in vitro fertilization (IVF) of porcine oocytes matured in vitro. This study was designed to examine whether embryo cryopreservation straws can be used to conduct IVF in porcine oocytes. The efficiency of this system was further compared with traditional microdrop IVF. Immature oocytes were aspirated from antral follicles and matured in vitro. After maturation, oocytes were inseminated either in straws or in microdrops with frozen-thawed boar spermatozoa. For straw IVF, sperm concentration and the presence of air columns between insemination segment and oil column were examined. Sperm-oocyte binding and cortical granules (CGs) before and after sperm penetration were examined by confocal microscopy. When various sperm concentrations were used for IVF in the straws with air columns, it was found that 5 x 106 cells/ml of sperm concentration was the optimal concentration; a high penetration rate (94.0%) and normal fertilization (oocytes with both male and female pronuclei) rate (38.2%) were obtained. Increasing sperm concentration to 10 x 106 cells/ml increased polyspermic penetration (61.9%) without affecting sperm penetration (86.9%). Reducing sperm concentration to 1 x 106 cells/ml reduced polyspermic penetration (25.6%), but sperm penetration rate (69.9%) was also reduced. When IVF was conducted in the straws with or without air columns, and in the microdrops, it was found that sperm penetration in the straws with air columns (96.5%) was significantly (p < 0.05) higher than that in the straws without air columns (81.7%) and in the microdrop (72.9%). However, the incidence of polyspermic penetration in the straws with air columns (34.2%) and without air columns (36.6%) was significantly (p < 0.05) lower than that (52.4%) in the microdrops. The number of spermatozoa bound to the oocytes was increased gradually in the straws but not in the microdrops in which more spermatozoa bound to the oocytes soon after insemination. CG exocytosis was more complete and faster in the oocytes inseminated in the straws than in the microdrops. These findings indicate that IVF of porcine oocytes in the straws provides a better condition in which more oocytes are fertilized normally than that in the microdrop IVF.

Bone regeneration or replacement has been proved to be one of the most effective methods available for the treatment of bone defects caused by different musculoskeletal disorders. However, the great contradiction between the large demand for clinical therapies and the insufficiency and deficiency of natural bone grafts has led to an urgent need for the development of synthetic bone graft substitutes. Bone tissue engineering has shown great potential in the construction of desired bone grafts, despite the many challenges that remain to be faced before safe and reliable clinical applications can be achieved. Graphene, with outstanding physical, chemical and biological properties, is considered a highly promising material for ideal bone regeneration and has attracted broad attention. In this review, we provide an introduction to the properties of graphene and its derivatives. In addition, based on the analysis of bone regeneration processes, interesting findings of graphene-based materials in bone regenerative medicine are analyzed, with special emphasis on their applications as scaffolds, membranes, and coatings in bone tissue engineering. Finally, the advantages, challenges, and future prospects of their application in bone regenerative medicine are discussed.

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Purpose/Objectives To identify factors associated with diagnostic delay after an incomplete or abnormal mammogram among women participating in a state mammography screening program. Research Approach Retrospective case-control design using bivariate and multivariate logistic regression analyses to explore the associations between age, race, ethnicity, marital status, breast cancer history, and self-reported breast symptoms and delay. Setting A statewide program of free screening mammography for women who are under- or uninsured. Participants 11,460 women enrolled in a free, statewide screening program from 2002–2006. Methodologic Approach Using the Tennessee Breast and Cervical Cancer Screening Program database, further analyses were conducted. Main Research Variables The outcome measure was delay in completion of all diagnostic tests and was defined as women who did not complete testing within 60 days. Findings: Thirty-seven percent of women required follow-up, and of a subset used in the analysis, 30% experienced delay of more than 60 days. Controlling for marital status, age, and breast cancer history, women who experienced delay were more likely to be African American versus Caucasian (odds ratio [OR] = 1.45, 95% confidence interval [CI] = 1.13, 1.85) or Hispanic (OR = 0.72, 95% CI = 0.55, 0.93) and to have self-reported breast symptoms (OR = 1.50, 95% CI = 1.27, 1.77). Conclusions In a sample of women with low income needing mammography follow-up, delay was associated with three intrapersonal variables, potentially reducing the effectiveness of mammography screening for women who were African American, or Hispanic, or had self-reported breast symptoms. Interpretation Nurses providing cancer screening examinations are uniquely positioned to assess the knowledge, beliefs, and resources of women using the program and to navigate women through barriers to completion. Knowledge of factors associated with delay is valuable for planning interventions and allocating program resources.

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