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Design of catalysts for Ni-catalyzed olefin polymerization predominantly focuses on ligand design rather than the activation process when attempting to achieve a broader scope of polyolefin micro- and macrostructures. Air-stable alkyl-or aryl-functionalized Ni II precatalysts were designed which eliminate the need of in situ alkylating processes and are activated solely by halide abstraction to generate the cationic complex for olefin polymerization. These complexes represent an emerging class of olefin polymerization catalysts, enabling the study of various cocatalysts forming either inner- or outer-sphere ion pairs. It is demonstrated that an organoboron cocatalyst activation produces a well-defined ion pair, which in contrast to ill-defined organoaluminum cocatalysts, can directly activate the complex by halide abstraction to yield comparatively higher molecular weight homo/copolymers. Under high ethylene pressure, broader branching densities and the gradual incorporation of short-chain branches were achieved, circumventing the need for elaborate ligand design and copolymerization with α-olefins. The underlying chain-walking mechanism and ion pair interactions were further elucidated by DFT calculations. A phenyl group on the bridging carbon functioned as a rotational barrier, producing higher molecular weight polymers compared to methyl-substituted analogs. Here, we provide a perspective to manipulate the iminopyridyl Ni II system, leveraging ion pair interactions and ligand design to govern polyolefin molecular weights and microstructures., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

The chemical structure of excipients molecularly mixed in an amorphous solid dispersion (ASD) has a significant impact on properties of the ASD including dissolution behavior, physical stability, and bioavailability. Polymers used in ASDs require a balance between hydrophobic and hydrophilic functionalities to ensure rapid dissolution of the amorphous dispersion as well as sustained supersaturation of the drug in solution. This work demonstrates the use of postpolymerization functionalization of poly(vinylpyridine) excipients to elucidate the impact of polymer properties on the dissolution behavior of amorphous dispersions containing posaconazole. It was found that N -oxidation of pyridine functionalities increased the solubility of poly(vinylpyridine) derivatives in neutral aqueous conditions and allowed for nanoparticle formation which supplied posaconazole into solution at concentrations exceeding those achieved by more conventional excipients such as hydroxypropyl methylcellulose acetate succinate (HPMCAS) or Eudragit E PO. By leveraging these functional modifications of the parent poly(vinylpyridine) excipient to increase polymer hydrophilicity and minimize the effect of polymer on pH, a new polymeric excipient was optimized for rapid dissolution and supersaturation maintenance for a model compound.

Block copolymers form the basis of the most ubiquitous materials such as thermoplastic elastomers, bridge interphases in polymer blends, and are fundamental for the development of high-performance materials. The driving force to further advance these materials is the accessibility of block copolymers, which have a wide variety in composition, functional group content, and precision of their structure. To advance and broaden the application of block copolymers will depend on the nature of combined segmented blocks, guided through the combination of polymerization techniques to reach a high versatility in block copolymer architecture and function. This review provides the most comprehensive overview of techniques to prepare linear block copolymers and is intended to serve as a guideline on how polymerization techniques can work together to result in desired block combinations. As the review will give an account of the relevant procedures and access areas, the sections will include orthogonal approaches or sequentially combined polymerization techniques, which increases the synthetic options for these materials.

Spironolactone improves cardiac structure, function and prognosis in patients with heart failure and delays the progression of cardiac fibrosis. However, the exact underlying mechanism of this process remains to be elucidated. The present study therefore aimed to explore the protective effect and underlying mechanism of the aldosterone receptor antagonist, spironolactone, on myocardial fibrosis in mice with experimental autoimmune myocarditis (EAM). The EAM model was induced in BALB/c mice via immunization with murine cardiac α-myosin heavy chain sequence polypeptides. The cardiac function of the mice was assessed using echocardiography and the levels of inflammatory cytokines were quantified using ELISA. E26 transformation-specific sequence-1 (Ets-1) expression was knocked down using lentivirus-mediated small interference RNA. Total collagen deposition was assessed using Masson's trichrome and Ets-1, TGF-β1, Smad2/3, collagen I and III protein expression levels were detected using immunohistochemistry and western blotting. MMP-2 and MMP-9 mRNA expression levels and activity was determined using reverse transcription-quantitative PCR and gelatin zymography, respectively. The results of the present study demonstrated that spironolactone significantly improved myocardium hypertrophy, diastolic cardiac function and decreased myocardial inflammation and collagen deposition induced by EAM. Spironolactone treatment significantly inhibited Ets-1 and smad2/3 phosphorylation. In addition, inhibition of Ets-1 reduced the expression and activity of MMP-2 and MMP-9 and decreased cardiac fibrosis in EAM mice. The results indicated that the improvement of myocardial fibrosis by spironolactone may be associated with the TGF-β1/Smad-2/3/Ets-1 signaling pathway in EAM mice., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Wang et al.)

A new species and a key to eleven species of of the genus Conophyma Zobovsky, 1898 from China is described in this paper. The new species Conophyma lini sp. nov. is similar to C. xiai Zhang et al, 2015, but differs from latter by: vertex longer, apex narrower; minimum width of interspace 1.6 times length in mesosternum; posterior margin of epiproct waved, with angular projection in the middle, furculae small and width of Epiphallus 2.5 times high. Type specimens are deposited in the Northwest Plateau Institute of Biology, Chinese Academy of Sciences, Xining, Qinghai 810001, China.

The introduction of m-xylyl substituents to α-diimine Pd II catalyst promotes living ethylene polymerization at room temperature and low pressure to yield high molecular weight polyethylene (PE) with low branching (<17/1000 C). m-Xylyl groups provide a highly effective blockage to the axial sites of the catalytic center and form a distorted sandwich geometry. The shielding prevents chain-transfer and easy accessibility of polar monomers, leading to a living polymerization. Conducting a light irradiation as part of the one-step metal-organic insertion light initiated radical (MILRad) process leads to diblock copolymers of ethylene and acrylates. Incorporation of different acrylate block sequences can significantly modify the mechanical and chemical properties of block copolymers which can be modulated to be a hard plastic, elastomer, or semi-amorphous polymer., (© 2021 Wiley-VCH GmbH.)

The first report of the red imported fire ant (RIFA), Solenopsis invicta Buren, in Taiwan was in the city of Taoyuan in 2003. The government has made great efforts to bring RIFA-infested areas under control. RIFA has gradually spread outward since its discovery, but it is still confined in northern Taiwan, in part due to the control efforts. RIFA is well established in densely populated environments (i.e., urban areas), causing damage to public utilities and significantly affects the inhabitants of Taiwan. Out of 10,127 human encounters with RIFA reported by the Plant Pest Information Management System in the Bureau of Animal and Plant Health Inspection and Quarantine, Council of Agriculture, Executive Yuan, 3819 (37.71%) persons were stung, with 834 (21.8%) persons exhibiting wheal-and-flare reaction (swelling and redness of the skin). Among the victims, 288 (7.5%) sought medical care, and about 21 (0.6%) developed severe cellulitis and urticaria. Unexpectedly, 2.8% (106) of the victims exhibited anaphylactic shock, which was higher than previously reported cases (1%). The high anaphylactic shock percentage was probably because most victims were elderly farmers or because Asian people have higher sensitivity to the RIFA sting. RIFA is well adapted to the environmental conditions in Taiwan, which makes it extremely difficult (if not impossible) to eradicate. The management of RIFA in the future should focus on lowering the speed of spread to mitigate possible dangers to the inhabitants. Six major challenges of RIFA management in Taiwan are also discussed.

The transverse-momentum-dependent (TMD) soft function is a key ingredient in QCD factorization of Drell-Yan and other processes with relatively small transverse momentum. We present a lattice QCD study of this function at moderately large rapidity on a 2+1 flavor CLS dynamic ensemble with a=0.098  fm. We extract the rapidity-independent (or intrinsic) part of the soft function through a large-momentum-transfer pseudoscalar meson form factor and its quasi-TMD wave function using leading-order factorization in large-momentum effective theory. We also investigate the rapidity-dependent part of the soft function-the Collins-Soper evolution kernel-based on the large-momentum evolution of the quasi-TMD wave function.

Polycystic ovary syndrome (PCOS) is one of the most common gynaecological endocrine disorders, and more than 60% of PCOS patients have varying degrees of insulin resistance (IR). The regulatory role of microRNAs (miRNAs) at post-transcriptional levels in human cumulus cells relating to IR in PCOS remains unclear. In this case-control study, 26 PCOS patients with IR (PCOS-IR) and 24 patients without IR (PCOS-control) were enrolled. We determined the differentially expressed miRNA and mRNA using next-generation sequencing technology, and these miRNAs and mRNAs were validated by quantitative real-time polymerase chain reaction (PCR). These miRNA regulating pathways (e.g., MAPK pathway) were analysed by bioinformatics analysis, and the Rap1b was demonstrated to be targeted by miR-612 based on quantitative real-time PCR, western blot and luciferase activity assay. A total of 59 known miRNAs and 617 differentially expressed genes were identified that differentially expressed between PCOS-IR and PCOS-control cumulus cells. Moreover, the potential regulating roles of miRNAs and their targeting genes in pathophysiology of IR and PCOS were analysed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotation, and several key processes were enriched, such as MAPK activity. Furthermore, Rap1b, a regulator of the MAPK pathway, was demonstrated to be suppressed directly by miR-612 in PCOS-IR cumulus cells based on negative expression correlation validation, dual luciferase activity assay and reduction of Rap1b expression after miR-612 mimics transfection. Our results suggested that miRNAs and their targeted pathways in ovarian cumulus cells may play important roles in the aetiology and pathophysiology of PCOS with IR.

The inherent differences in reactivity between activated and non-activated alkenes prevents copolymerization using established polymer synthesis techniques. Research over the past 20 years has greatly advanced the copolymerization of polar vinyl monomers and olefins. This Review highlights the challenges associated with conventional polymerization systems and evaluates the most relevant methods which have been developed to "bridge the gap" between polar vinyl monomers and olefins. We discuss advancements in heteroatom tolerant coordination-insertion polymerizations, methods of controlling radical polymerizations to incorporate olefinic monomers, as well as combined approaches employing sequential polymerizations. Finally, we discuss state-of-the-art stimuli-responsive systems capable of facile switching between catalytic pathways and provide an outlook towards applications in which tailored copolymers are ideally suited., (© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

We present a state-of-the-art calculation of the isovector quark-helicity Bjorken-x distribution in the proton using lattice-QCD ensembles at the physical pion mass. We compute quasidistributions at proton momenta P&#95;{z}∈{2.2,2.6,3.0}  GeV on the lattice and match them systematically to the physical parton distribution using the large-momentum effective theory. We reach an unprecedented precision through high statistics in simulations, large-momentum proton matrix elements, and control of excited-state contamination. The resulting distribution with combined statistical and systematic errors is in agreement with the latest phenomenological analysis of the spin-dependent experimental data, in particular, Δu[over ¯](x)>Δd[over ¯](x).

Problem: The role of vitamin D (VD) in IVF outcome and immune parameters has not been elucidated well., Method of Study: Women undergoing IVF treatment with GnRH agonist (Agonist) and progestin-primed ovarian stimulation (PPOS) protocols were divided into VD lower (VDL, 25(OH)VD ≤20 ng/mL) and VD higher (VDH, 25(OH)VD >20 ng/mL) groups. Follicular fluid (FF) VD level, IVF outcomes, and peripheral blood immunophenotypes by flow cytometry were analyzed., Results: FF VD levels of the whole subjects were positively correlated with peripheral blood VD level (r = 0.86, P < 0.001). The number of mature oocytes and the blastocyst formation rate were significantly higher in women with VDH group as compared with those of VDL group in both Agonist and PPOS groups (P < 0.05, respectively). In women with PPOS protocol, peripheral blood NK and B-cell proportions and T helper/T cytotoxic (Th/Tc) cell ratios of VDL group were significantly higher than those of VDH group (P < 0.05, respectively). In women with Agonist protocol, peripheral blood B-cell proportion and Th/Tc ratios of VDL group were significantly higher than those of VDH group (P < 0.05, respectively)., Conclusion: VD level is associated with IVF outcomes possibly derived by T-cell immunity, particularly Th/Tc ratios., (© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Intrauterine adhesions (IUA) are associated with the loss of stem cells in the endometrium. Menstrual blood‑derived stem cells (MenSCs) can be isolated from the menstrual blood and differentiated into endometrial cells. To check the transplantation feasibility of MenSCs for the treatment of severe IUA, MenSCs were isolated from menstrual blood, cultured in Dulbecco's modified Eagle's medium (DMEM), identified by immunocytochemistry and flow cytometry, differentiated into endometrial cells in vitro, and finally transplanted into the axillary subcutaneous tissue of non‑obese diabetic/severe combined immunodeficiency (NOD‑SCID) mice to create endometrial tissue. Additionally, the cloning efficiency and POU domain class 5 transcription factor 1 (OCT‑4) positivity of MenSCs from patients with severe IUA were compared with those from healthy women. Immunocytochemistry and flow cytometry results showed that 95.1±0.8% cells were OCT‑4‑positive, 0.9±0.4% were cluster of differentiation (CD)45‑positive, 1.8±0.9% were STRO‑1‑positive and 1.0±0.4% were human leukocyte antigen‑antigen D related‑positive. Following differentiation in vitro, the results of immunocytochemistry, reverse transcription‑polymerase chain reaction and western blot analysis showed that the expression of cytokeratin (CK) and vimentin (VIM) was increased in MenSCs compared with that in control subjects. Subsequent to transplantation in mice administered with sequential 17β‑estradiol and progesterone, CK, VIM, estrogen receptor and progesterone receptor were expressed in the transplantation regions, suggesting that MenSCs could differentiate into endometrial tissues in vivo. The cloning efficiency and OCT‑4 positivity of MenSCs from patients with severe IUA was significantly decreased. In conclusion, to the best of our knowledge, this is the first study in which MenSCs could differentiate into endometrial cells in vitro and create endometrial tissue in NOD‑SCID mice in vivo, with impaired cloning efficiency and OCT‑4 expression of MenSCs from patients with IUA. This study will provide a theoretical basis for the treatment of IUA with MenSCs.

Utilization of triplet excitons plays a key role in obtaining highly efficient quantum-dot light-emitting diodes (QD-LEDs). However, to date, only phosphorescent materials have been implemented to harvest triplet excitons in QD-LEDs. In this work, we introduced a thermally activated delayed fluorescence (TADF) emitter, 4,5-di(9H-carbazol-9-yl)phthalonitrile (2CzPN), doped into poly(N-vinylcarbazole) (PVK) as an exciton harvester in red QD-LEDs by solution processing. As a result, electrons leaking to the PVK layer will be trapped by 2CzPN to form long-lifetime TADF excitons in the 2CzPN:PVK layer, and then this harvested exciton energy can be effectively transferred to the adjacent QDs by the Förster resonance energy-transfer process. The fabricated red CdSe/CdS core/shell QD-LEDs show a maximum luminescence efficiency of 17.33 cd/A and longer lifetime. Our results demonstrate that the TADF sensitizer would be a promising candidate to develop highly efficient QD-LEDs.

Problem: We aim to investigate a possible role of IL-7/IL-7R signaling pathway in recurrent pregnancy losses (RPL)., Material and Methods: Using the abortion-prone (AP) and non-abortion-prone (NP) mice model, fetal resorption rates (FRR), Th17 and Treg cells-related factors, and the effect of IL-7 and IL-7R antagonist were investigated by flow cytometry, quantitative real-time PCR, and immunohistochemistry. IL-7 and IL-7R expressions in human decidua were investigated by immunohistochemistry., Results: In the AP mice, IL-7R antagonist treatment significantly decreased FRR by downregulating Th17 and upregulating Treg-related factors. When the NP mice were treated with IL-7, FRR was significantly increased by upregulating Th17 and downregulating Treg-related factors. In decidual stromal cells of women with RPL, increased IL-7 and decreased IL-7R expressions were present when compared to normal controls., Conclusion: IL-7/IL-7R signaling pathway plays a possible role in RPL by upregulating Th17 immunity, meanwhile downregulating Treg immunity. Regulation of IL-7/IL-7R may be a new therapeutic strategy for RPL., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

A new scalar boson which couples to the muon and proton can simultaneously solve the proton radius puzzle and the muon anomalous magnetic moment discrepancy. Using a variety of measurements, we constrain the mass of this scalar and its couplings to the electron, muon, neutron, and proton. Making no assumptions about the underlying model, these constraints and the requirement that it solve both problems limit the mass of the scalar to between about 100 keV and 100 MeV. We identify two unexplored regions in the coupling constant-mass plane. Potential future experiments and their implications for theories with mass-weighted lepton couplings are discussed.

Recently, post-transcriptional gene regulation by microRNAs (miRNAs) has been reported to play a key role during ovary development and differentiation. However, there are no published studies identifying miRNA profiles of human ovarian tissues directly using next-generation sequencing technology. In the human ovary, a total of 762 known and 21 novel human miRNAs were detected, indicating that human ovaries have a complex population of small RNAs. To confirm the miRNA profile in human ovaries, quantitative real-time polymerase chain reaction was used to validate the expression of known miRNAs and novel miRNAs. The potential regulating roles of miRNA in physiological function of ovaries were analyzed by gene ontology and Kyoto encyclopedia of genes and genomes pathway annotation, and several important processes were identified to be targeted by the most abundantly expressed miRNAs, for example, antral ovarian follicle growth, ovarian follicle rupture, and fertilization. Our current findings extend the knowledge of the regulatory role of miRNAs and their targeted processes in human ovaries, suggesting that miRNAs play important roles in development and physiological function of ovaries. In this study, we provide a useful resource for further research of the regulatory role of miRNAs in the ovaries, which may also provide novel candidates for molecular biomarkers or treatment targets in the research of female infertility.

We previously synthesized a chondroitin sulfate-graft-poly(ε-caprolactone) copolymer (H-CP) with a high content of poly(ε-caprolactone) (18.7 mol%), which self-assembled in water into a rod-like micelle to encapsulate hydrophobic camptothecin (CPT) in the core (micelle/CPT) for tumor-targeted drug delivery. As a result of the recognition of the micelle by CD44, the micelle/CPT entered CRL-5802 cells efficiently and released CPT efficaciously, resulting in higher tumor suppression than commercial CPT-11. In this study, H1299 cells were found to have a higher CD44 expression than CRL-5802 cells. However, the lower CD44-expressing CRL-5802 cells had a higher percentage of cell death and higher cellular uptake of the micelle/CPT than the higher CD44-expressing H1299 cells. Examination of the internalization pathway of the micelle/CPT in the presence of different endocytic chemical inhibitors showed that the CRL-5802 cells involved clathrin-mediated endocytosis, which was not found in the H1299 cells. Analysis of the cell cycle of the two cell lines exposed to the micelle/CPT revealed that the CRL-5802 cells arrested mainly in the S phase and the H1299 cells arrested mainly in the G2-M phase. A consistent result was also found in the evaluation of γ-H2AX expression, which was about three-fold higher in the CRL-5802 cells than in the H1299 cells. A near-infrared dye, IR780, was encapsulated into the micelle to observe the in vivo biodistribution of the micelle/IR780 in tumor-bearing mice. The CRL-5802 tumor showed a higher fluorescence intensity than the H1299 tumor at any tracing time after 1 h. Thus we tentatively concluded that CRL-5802 cells utilized the clathrin-mediated internalization pathway and arrested in the S phase on exposure to the micelle/CPT; all are possible reasons for the better therapeutic outcome in CRL-5802 cells than in H1299 cells.

Objective: Krüppel-like factor 2 (KLF2) is a transcription factor that regulates endothelial function and atorvastatin can stabilize atherosclerotic plaque and inhibit inflammation on endothelial cells by attenuating the role of cytokines. The aim of this study is to investigate the effect of high glucose (HG) on KLF2 expression in human umbilical vein endothelial cells (HUVECs) and the underlying mechanisms., Methods: HUVECs were isolated from the human umbilical cords from normal pregnancies and exposed to medium containing 25.5 mM D-glucose for 24 hours as the HG induction model (HG group). In the HG plus atorvastatin groups or KLF2 gene transduction, the medium then was collected for the nitric oxide (NO) assay and the cells were harvested for Western blot and for the real-time polymerase chain reaction to observe the expression of KLF2, vascular cell adhesion molecule (VCAM)-1, intercellular adhesion molecule (ICAM)-1, total and phosphorylated endothelial NO synthase (eNOS), p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK)1/2, caspase-3 and cleaved caspase-3 and the role of the p38MAPK and ERK1/2 intracellular signal pathway. The cells' apoptosis was analyzed by flow cytometry., Results: HG dose-dependently increased apoptosis. The presence of HG inhibited the expression of KLF2 mRNA and protein in HUVECs and atorvastatin treatment increased KLF2 expression, thus counteracted HG-induced suppression of KLF2 expression, and overexpression of KLF2 might protect the cells from apoptosis. HG increased the expression of VCAM-1, ICAM-1, but decreased the nitric oxide release and the expression of p-eNOs/eNos in HUVECs. However, atorvastatin reversed these changes and also attenuated high-glucose induced p38 MAPK and ERK1/2 phosphorylation., Conclusions: HG suppressed the KLF2 expression in HUVECs. The suppression was counteracted by atorvastatin treatment, probably via attenuating the activation of the signal pathyway p38 MAPK and ERK1/2.

Endometriosis, a pathological condition in which the endometrium grows outside the uterus, is one of the most common causes of female infertility; it is diagnosed in 25-40% of infertile women. The mechanism by which endometriosis affects the fertility of females remains largely unknown. We examined the ultrastructure of oocytes from patients with minimal or mild endometriosis and control females undergoing in vitro fertilization (IVF) treatment by transmission electron microscopy (TEM) to investigate the physiological significance of oocyte quality for patients with minimal or mild endometriosis. The TEM results revealed that the oocytes from women with minimal or mild endometriosis exhibited abnormal mitochondrial structure and decreased mitochondria mass. Quantitative real time PCR analysis revealed that the mitochondrial DNA copy number was significantly reduced in the oocytes from women with minimal or mild endometriosis compared with those of the control subjects. Our results suggest that decreased oocyte quality because of impaired mitochondrial structure and functions probably an important factor affecting the fertility of endometriosis patients.

MicroRNA-128 (miR-128) is an attractive therapeutic molecule with powerful glioblastoma regulation properties. However, miR-128 lacks biological stability and leads to poor delivery efficacy in clinical applications. In our previous study, we demonstrated two effective transgene carriers, including polyethylenimine (PEI)-decorated superparamagnetic iron oxide nanoparticles (SPIONs) as well as chemically-conjugated chondroitin sulfate-PEI copolymers (CPs). In this contribution, we report optimized conditions for coating CPs onto the surfaces of SPIONs, forming CPIOs, for magneto-gene delivery systems. The optimized weight ratio of the CPs and SPIONs is 2 : 1, which resulted in the formation of a stable particle as a good transgene carrier. The hydrodynamic diameter of the CPIOs is ∼136 nm. The gel electrophoresis results demonstrate that the weight ratio of CPIO/DNA required to completely encapsulate pDNA is ≥3. The in vitro tests of CPIO/DNA were done in 293 T, CRL5802, and U87-MG cells in the presence and absence of an external magnetic field. The magnetofection efficiency of CPIO/DNA was measured in the three cell lines with or without fetal bovine serum (FBS). CPIO/DNA exhibited remarkably improved gene expression in the presence of the magnetic field and 10% FBS as compared with a gold non-viral standard, PEI/DNA, and a commercial magnetofection reagent, PolyMag/DNA. In addition, CPIO/DNA showed less cytotoxicity than PEI/DNA and PolyMag/DNA against the three cell lines. The transfection efficiency of the magnetoplex improved significantly with an assisted magnetic field. In miR-128 delivery, a microRNA plate array and fluorescence in situ hybridization were used to demonstrate that CPIO/pMIRNA-128 indeed expresses more miR-128 with the assisted magnetic field than without. In a biodistribution test, CPIO/Cy5-DNA showed higher accumulation at the tumor site where an external magnet is placed nearby.

Context: Polycystic ovary syndrome (PCOS), a complex and heterogeneous endocrine condition, is characterized by polycystic ovaries, hyperandrogenism, insulin resistance and chronic anovulation. Cumulus granulosa cells surrounding the oocyte are involved in different aspects of PCOS pathology. Several studies suggested that miRNAs play an important regulatory role at the post-transcriptional level in cumulus granulosa cells., Objective: Our objective was to describe the altered miRNA expression profiles and miRNA targeted signaling pathways in PCOS., Design: Case-control study that involved 21 women with PCOS and 20 women without the disease (controls). The miRNA expression profiles of human cumulus granulosa cells were determined using next generation sequencing by Illumina Hiseq 2000. The differentially expressed miRNAs and novel miRNAs were validated by quantitative real-time PCR. The Notch3 and MAPK3 were demonstrated to be targeted by miR-483-5p based on quantitative real-time PCR, western blot and luciferase activity assay., Results: Compared with controls, a total of 59 known miRNA were identified that differentially expressed in PCOS cumulus granulosa cells, including 21 miRNAs increase and 38 miRNAs decrease. Moreover, the novel miRNAs were predicted in PCOS and control cumulus granulosa cells. The potential regulating roles of miRNA in pathophysiology of PCOS were analyzed by GO and KEGG pathway annotation, and several important processes were identified to be targeted by the differentially expressed miRNAs, such as Notch signaling, regulation of hormone, and energy metabolism. Furthermore, Notch3 and MAPK3, the members of Notch signaling and ERK-MAPK pathway, were demonstrated to be regulated by miR-483-5p based on negative expression correlation validation and detection of Notch3/MAPK3 expression after miR-483-5p mimics transfection. Dual luciferase activity assay suggested that Notch3 and MAPK3 were directly targeted by miR-483-5p., Conclusion: Our data suggested that miRNAs and their targeted pathways (e.g. Notch signaling pathway) play important roles in the etiology and pathophysiology of PCOS, and provides novel candidates for molecular biomarkers or treatment targets in the research of female infertility associated with PCOS., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Objective: Sitagliptin, a dipeptidyl peptidase IV (DPP-Ⅳ) inhibitor, has a biological role in improving the serum levels of glucagon-like peptide 1 (GLP-1). Hence, we sought to determine the effect of sitagliptin on myocardial inflammation, collagen metabolism, lipid content and myocardial apoptosis in diabetic rats., Materials and Methods: The type 2 diabetic rat model was induced by low-dose streptozotocin and a high-fat diet. Characteristics of diabetic rats were evaluated by electrocardiography, echocardiography and blood analysis. Cardiac inflammation, fibrosis, cardiomyocyte density, lipid accumulation, and receptor-interacting protein kinase 3 (RIP3) level, related to apoptosis, were detected by histopathologic analysis, RT-PCR and western blot analysis to evaluate the effects of sitagliptin on myocardial remodeling of the left ventricle., Results: Diabetic rats showed myocardial hypertrophy or apoptosis, inflammation, lipid accumulation, myocardial fibrosis, elevated collagen content, RIP3 overexpression, and left-ventricular dysfunction. Sitagliptin could reverse the overexpression of RIP3 and alleviate cellular apoptosis in myocardial tissues. It could significantly improve left-ventricular systolic pressure and +dp/dt max, reduce the E/E' ratio, left ventricular end diastolic pressure, -dp/dt max and Tau in diabetic rats., Conclusions: Sitagliptin might have a myocardial protective effect by inhibiting apoptosis, inflammation, lipid accumulation and myocardial fibrosis in diabetic rats, for a potential role in improving left-ventricular function in diabetes., (Copyright © 2014 Japanese Pharmacological Society. Production and hosting by Elsevier B.V. All rights reserved.)

Adiponectin (APN) is an important anti‑atherogenic adipocytokine. The aim of the present study was to investigate the role of adiponectin in atherosclerotic plaque formation and clarify its mechanisms. An atherosclerosis model was induced by in vivo perivascular constrictive silica collar placement on the left common carotid arteries in male apolipoprotein E‑deficient (ApoE‑/‑) mice. All of the mice were fed a high‑fat diet, and divided into phosphate‑buffered saline, adenovirus (Ad)‑β‑galactosidase and Ad‑APN treatment groups. Compared with treatment of Ad‑β‑gal or PBS, Ad‑APN treatment markedly reduced inducible nitric oxide synthase (iNOS) protein expression, decreased in nitric oxide/superoxide production, blocked peroxynitrite formation and reversed the progression of atherosclerotic lesions. Adiponectin may be a natural molecule that reduces atherosclerosis by inhibiting iNOS and consequently diminishing oxidative/nitrative stress.

Objective: Hypoxia inducible factor-1α (HIF-1α) regulates many genes involved in angiogenesis during embryonic development. Epidermal growth factor-like domain 7 (Egfl7) is a specific marker for human arterial endothelial cells that are in an activated state of proliferation, migration, and remodeling. This study evaluates the intricate relationship between HIF-1α and Egfl7 under both hyperoxia and hypoxia states., Methods: The neonatal mice were exposed to either hyperoxia or hypoxia in order to detect the pulmonary and cardiac Egfl7 messenger RNA (mRNA) or protein expression regulated by oxygen tension in vivo by reverse transcriptase polymerase chain reaction or immunohistochemistry staining. Egfl7 expression in HIF-1α null pulmonary endothelial cells in hypoxia conditions and effects of overexpression or knockdown of HIF-1α on Egfl7 expression in human umbilical vein endothelial cells would be clarified in vitro by reverse transcriptase polymerase chain reaction and Western blot, respectively., Results: Hyperoxia exposure significantly reduced Egfl7 expression in neonatal mice lungs by 36% compared with age-matched normoxia control mice (P < 0.05, n = 6). The pulmonary Egfl7 transcription levels were increased by 1.7- and 1.9-fold in 24 hours and by day 8 in hypoxia groups compared with the normoxia control values (P < 0.05, n = 6). The cardiac Egfl7 mRNA expression was significantly increased by 4.5-fold in the day 8 group compared with the normoxia control values (P < 0.05, n = 6). The expression of Egfl7 decreased significantly in the HIF-1α(-/-) endothelial cells (ECs), which was only 26% of wild-type HIF-1α(+/+) ECs (P < 0.05, n = 3). Hypoxia caused a mild but significant increase of Egfl7 expression in HIF-1α(+/+) ECs (P < 0.05). In vitro, overexpression of HIF-1α enhanced Egfl7 expression, whereas knockdown of HIF-1α reduced Egfl7 expression., Conclusions: Overexpression of HIF-1α enhanced Egfl7 expression, whereas knockdown of HIF-1α reduced Egfl7 expression. Egfl7 could be a HIF-1α responsive gene regulated by oxygen tension.

In this study, we aimed to detect the effects of the soluble epoxide hydrolase (sEH) inhibitor 12-(3-adamantan-1-yl-ureido)-dodecanoic acid (AUDA) on atherosclerotic diseases and to explore its mechanism. The atherosclerosis animal model was constructed by ApoE-/- mice. To determine the optimal therapeutic concentration of AUDA, different concentrations of AUDA were infused into ApoE-/- mice, with controls receiving infusions of normal saline alone. Mouse body weight and serum total cholesterol, triglyceride, LDL and HDL levels were measured. The western blotting (WB) method was used to detect the expression of TLR4 and NFKB in the aortic wall of the AUDA-treated and control mice. After the animals were sacrificed, we performed Oil Red O staining of the aortic sinus atherosclerotic plaque area followed by quantitative analysis of the aortic atherosclerotic plaque size and the percentage of lumen area in the two groups of mice. The expression levels of inflammatory cytokines, adhesion molecules and chemokines in the AUDA group were significantly decreased compared to the saline-treatment group (P < 0.05). The optimal AUDA concentration was found to be 0.35 ml/mg. AUDA significantly inhibited the expression of TLR4 and NFκB in ApoE-/- mouse aortas and reduced the aortic sinus plaque area of the ApoE-/- mouse group (P < 0.05). In conclusion, AUDA can regulate blood lipid balance, which may be one of the mechanisms for its protective effects on the cardiovascular system.

During folliculogenesis, cumulus cells surrounding the oocyte differentiate into corona radiata cells (CRCs) and cumulus oophorus cells (COCs), which are involved in gonadal steroidogenesis and the development of germ cells. Several studies suggested that microRNAs (miRNAs) play an important regulatory role at the post-transcriptional level in cumulus cells. However, comparative miRNA profiles and associated processes in human CRCs and COCs have not been reported before. In this study, miRNA profiles were obtained from CRCs and COCs using next generation sequencing in women undergoing controlled ovarian stimulation for IVF. A total of 785 and 799 annotated miRNAs were identified in CRCs and COCs, while high expression levels of six novel miRNAs were detected both in CRCs and in COCs. In addition, different expression patterns in CRCs and COCs were detected in 72 annotated miRNAs. To confirm the miRNA profile in COCs and CRCs, quantitative real-time PCR was used to validate the expression of annotated miRNAs, differentially expressed miRNAs, and novel miRNAs. The miRNAs in the let-7 family were found to be involved in the regulation of a broad range of biological processes in both cumulus cell populations, which was accompanied by a large amount of miRNA editing. Bioinformatics analysis showed that amino acid and energy metabolism were targeted significantly by miRNAs that were differentially expressed between CRCs and COCs. Our work extends the current knowledge of the regulatory role of miRNAs and their targeted pathways in folliculogenesis, and provides novel candidates for molecular biomarkers in the research of female infertility.

Background: Several types of T cells have been associated with the pathogenesis of unexplained recurrent spontaneous abortion (URSA), including Th1/Th2/Th17/Tregs cell. It has been appreciated that immunotherapy with paternal or third party lymphocytes is an effective method of treatment for URSA patients. The balance of Th1/Th2 cells could be maintained and an increase of Treg cells would be beneficial after immunotherapy; however, the mechanism by which the Th17/Treg balance affects URSA has not yet been fully elucidated., Methods: Here, we used flow cytometry, liquid chip technology and quantitative real-time PCR (qPCR) methods to characterize Th17/Treg cell populations after immunotherapy. We found that after immunotherapy in URSA patients, the percentage of Th17 cells decreased and the percentage of Treg cells in peripheral blood mononuclear cells (PBMC) increased, as detected by flow cytometry., Results: Immunotherapy may induce a decrease in the Th17/Treg ratio and the Treg bias, which may be beneficial for the maintenance of pregnancy. The expression level of ROR gamma t, a transcription factor found in Th17 cells, decreased and the expression of the Treg-specific transcription factor Foxp3 increased in peripheral blood as detected by qPCR. Immunotherapy may induce a decrease in the ratio of ROR gamma t to Foxp3 and a Treg cell bias, which would be beneficial for pregnancy maintenance. The secretion of the Treg-associated cytokine TGF-beta, as well as Th2 cytokines, was increased in serum, while the secretion of Th17-associated cytokine IL-17A and Th1 cytokine production was decreased. The Th1/Th2 cytokine ratio significantly decreased. Similarly, the Th17/Treg ratio significantly decreased in the total patient after immunotherapy., Conclusions: These results indicate that in patients with URSA, immunotherapy with mononuclear cells derived from the baby's father could affect both Th1/Th2 and Th17/Treg balance, and we found that the Th2 and Treg bias would be beneficial for pregnancy, which may lead to a balancing of the Th17/Treg ratio in URSA patients after immunotherapy.

The aim of this study was to examine the effects of vitrification with autologous follicular fluid (AFF) supplemented with ethylene glycol (EG) and sucrose on human oocytes with corona radiata. A total of 182 human oocytes with corona radiata from fifteen infertile patients were vitrified using either equilibration solutions (ES) and vitrification solution (VS) consisting of AFF, EG and sucrose (AFF group, n=67) or commercial ES and VS (control group, n=115). All oocytes were thawed in the next cycle, with surviving oocytes being inseminated by conventional IVF. The clinical outcome of vitrified-warmed oocytes by both vitrification methods was analysed retrospectively. In the AFF group, six patients received embryo transfer, with three couples taking four healthy babies home. In the control group, nine patients received embryo transfer, with four couples taking five healthy babies home. There was no significant difference in the survival rate (91.0 vs 92.2%), two pronuclei (2PN) fertilisation rate (73.8 vs 73.6%), cleavage rate (100 vs 100%), top-quality embryo rate (62.2 vs 59.2%), clinical pregnancy rate (50.0 vs 44.4%), implantation rate (33.3 vs 25%) or take-home baby rate (50.0 vs 44.4%) between the AFF group and the control group, respectively. These results show that AFF supplemented with EG and sucrose is an efficient, cost-effective cryoprotectant for human oocyte cryopreservation. A corona radiata on vitrified-warmed oocytes retains the oocytes' fertilisation capability in conventional IVF.

The modern European climatic regime is peculiar, due to its unitary winter but diverse summer climates and a pronounced Mediterranean climate in the south. However, little is known on its evolution in the deep time. Here we reconstruct the European summer climate conditions in the Tortonian (11.62-7.246 Ma) using plant fossil assemblages from 75 well-dated sites across Europe. Our results clearly show that the Tortonian Europe mainly had humid to subhumid summers and no arid climate has been conclusively detected, indicating that the summer-dry Mediterranean-type climate has not yet been established along most of the Mediterranean coast at least by the Tortonian. More importantly, the reconstructed distribution pattern of summer precipitation reveals that the Tortonian European must have largely been controlled by westerlies, resulting in higher precipitation in the west and the lower in the east. The Tortonian westerly wind field appears to differ principally from the trade wind pattern of the preceding Serravallian (13.82-11.62 Ma), recently deduced from herpetofaunal fossils. Such a shift in atmospheric circulation, if ever occurred, might result from the development of ice caps and glaciers in the polar region during the Late Miocene global cooling, the then reorganization of oceanic circulation, and/or the Himalayan-Tibetan uplift.

Chondroitin sulfate-g-poly(ε-caprolactone) (CP) copolymers were synthesized via atom transfer radical addition (ATRA). The CP copolymers self-assembled into micelles in water, and the micelles could be used to encapsulate a hydrophobic anticancer drug, camptothecin (CPT), in the core for tumor targeting delivery. The physicochemical properties of the micelles and CPT-loaded micelles were thoroughly characterized. For the in vitro test, the CPT release, the protection of the lactone ring of CPT from hydrolysis and the cellular uptake of CPT were studied. The cell-killing and apoptosis-inducing effects using the CPT-loaded micelles were significantly better than using free CPT against CRL-5802 cells. The micellar internalization into CRL-5802 cells was primarily via CD44 and clathrin dual-mediated endocytosis. For the in vivo test, the therapeutic efficacy of the CPT-loaded micelles was studied in a non-small-cell lung cancer xenograft animal model. The CPT-loaded micelles showed good inhibition in tumor growth as compared with a commercial product, CPT-11, in CRL-5802 tumor-bearing mice. The in vitro and in vivo data suggested the CP-based micelles are promising anticancer drug vehicles for lung cancer targeting.

The morphological assessment of oocytes is important for embryologists to identify and select MII oocytes in IVF/ICSI cycles. Dysmorphism of oocytes decreases viability and the developmental potential of oocytes as well as the clinical pregnancy rate. Several reports have suggested that oocytes with a dark zona pellucida (DZP) correlate with the outcome of IVF treatment. However, the effect of DZP on oocyte quality, fertilization, implantation, and pregnancy outcome were not investigated in detail. In this study, a retrospective analysis was performed in 268 infertile patients with fallopian tube obstruction and/or male factor infertility. In 204 of these patients, all oocytes were surrounded by a normal zona pellucida (NZP, control group), whereas 46 patients were found to have part of their retrieved oocytes enclosed by NZP and the other by DZP (Group A). In addition, all oocytes enclosed by DZP were retrieved from 18 patients (Group B). No differences were detected between the control and group A. Compared to the control group, the rates of fertilization, good quality embryos, implantation and clinical pregnancy were significantly decreased in group B. Furthermore, mitochondria in oocytes with a DZP in both of the two study groups (A and B) were severely damaged with several ultrastructural alterations, which were associated with an increased density of the zona pellucida and vacuolization. Briefly, oocytes with a DZP affected the clinical outcome in IVF/ICSI cycles and appeared to contain more ultrastructural alterations. Thus, DZP could be used as a potential selective marker for embryologists during daily laboratory work.

Background: Anti-nuclear antibodies (ANA) are suspected of having relevance to adverse reproductive events., Methods: This study aims to investigate the potential effect of ANA on IVF/ICSI outcome and the therapeutic role of prednisone plus low-dose aspirin (P + A) adjuvant treatment in ANA + patients. The first IVF/ICSI cycles without P + A of sixty-six ANA + women were enrolled as the ANA + group, and the 233 first IVF/ICSI cycles of matched ANA- women served as the ANA- group. The ANA + group was divided into the Titre < =1:320 subgroup and the Titre > 1:320 subgroup. Twenty-one ANA + women with adverse outcomes in their first cycles (ANA + cycles without P + A) received P + A adjuvant treatment for three months before the second IVF/ICSI cycle (ANA + cycles with P + A). The clinical characteristics and the IVF/ICSI outcomes were compared, respectively, between 1) the ANA + group and the ANA- group, 2) the Titre < =1:320 subgroup and the Titre > 1:320 subgroup, and 3) the ANA + cycles without P + A and the ANA + cycles with P + A., Results: No significant differences were observed between each of the two-group pairs in the clinical characteristics. The ANA + group exhibited significantly lower MII oocytes rate, normal fertilisation, pregnancy and implantation rates, as well as remarkably higher abnormal fertilisation and early miscarriage rates. The Titre < =1:320 subgroup's IVF/ICSI outcomes were as poor as those of the Titre > 1:320 subgroup. After the P + A adjuvant treatment, the number of two pro-nuclei, perfect embryos and available embryos, and the implantation rate increased significantly., Conclusions: These observations suggest that ANA could exert a detrimental effect on IVF/ICSI outcome that might not be titre-dependent, and P + A adjuvant treatment could be useful for ANA + patients. This hypothesis should be verified in further prospective randomised studies.

Objective: More attention is being paid to the relationship between kidney dysfunction and cardiovascular events Characteristic features include renal dysfunction, left ventricular (LV) and left atrial (LA) enlargement. The aim of this study is to evaluate the relationships between circulating levels of β2-microglobulin (β2-m) and cystatin C and left atrial size in patients with coronary artery disease., Materials and Methods: We recruited 300 patients who presented with chest tightness or chest pain and subsequently underwent coronary angiography. Of these, 202 patients were diagnosed with coronary artery disease (CAD) and 98 patients without CAD (non-CAD). Laboratory measurements included liver, kidney function (urea nitrogen, creatinine, β2-m and cystatin C), fasting glucose, and lipid analysis. CrCl were calculated according to Cockroft-Gault formula. Echocardiology was used to evaluate the cardiac structure and function., Results: Significant differences of β2-m and cystatin C exist and no difference of creatinine and CrCl existed between the two groups. LA diameters were positively related to circulating levels of β2-m in the CAD group (r = 0.452, p < 0.001) and non-CAD group (r = 0.360, p < 0.001), and the similar relationships between LAD and circulating levels of cystatin C in the CAD group (r = 0.302, p < 0.001) and non-CAD group (r = 0.243, p = 0.016). LA diameters were negatively related to CrCl in both groups. After multivariate logistic regression analysis, the data indicated that the independent cardiovascular risk factors of LA enlargement for the patients with CAD were age, BMI, systolic blood pressure, LV mass, LVEDD, E/A, Em/Am, CrCl, and circulating levels of β2-m (OR = 1.630, 95% CI: 1.115 - 2.384, p = 0.012), cystatin C (OR = 4.504, 95% CI: 1.478 - 13.726, p = 0.008)., Conclusions: A linear correlation exists between circulating levels of β2-m or cystatin C and LA diameters. Higher circulating levels of β2-m or cystatin C are independent cardiovascular risk factors of LA enlargement in patients with CAD, and could be a link between the kidney and the heart.

Objective: Dickkopf-1 (DKK-1), a major regulator of the Wnt pathway, plays an important role in cardiovascular disease. However, no study has evaluated the association of DKK-1 and acute coronary syndrome (ACS). We investigated this association and whether the Global Registry of Acute Coronary Events (GRACE) hospital-discharge risk score predicting major adverse cardiac events (MACE) can be improved by adding the DKK-1 value., Methods: We enrolled 291 patients (46 with ST-segment elevation myocardial infarction [STEMI] and 245 with non-ST elevated ACS [NSTE-ACS]) who were divided into groups by tertiles of baseline plasma DKK-1 level measured by ELISA. The GRACE risk score was calculated and predictive value alone and together with DKK-1 and/or high-sensitivity C-reactive protein (hs-CRP) level were assessed, respectively., Results: Compared with patients with NSTE-ACS, those with STEMI had higher plasma DKK-1 level at baseline (P = 0.006). Plasma DKK-1 level was correlated with hs-CRP level (r = 0.295, P<0.001) and was greater with high than intermediate or low GRACE scores (P = 0.002 and P<0.001, respectively). We found 44 (15.1%) MACEs during a median 2-year follow-up. DKK-1 levels were higher for patients with than without events (P<0.001). The rate of MACE increased with increasing DKK-1 level (P<0.001). The area under the receiver operating characteristic curve for GRACE score with MACE was 0.524 and improved to 0.791 with the addition of hs-CRP level, 0.775 with the addition of DKK-1 level and 0.847 with both values added., Conclusions: DKK-1 is an independent predictor of long-term MACE of patients with ACS. The long-term predictive ability of post-discharge GRACE score may be enhanced by adding DKK-1 level.

Objective: To study the population dynamics of aphid on Lonicera macranthoides and their natural enemy in Xiushan and control method of pesticide so as to provide scientific basis for its integrated pests management (IPM)., Method: The field investigation and the field controlling trial were carried out for the research., Result: Semiaphis heraclei was the dominant species among L. macranthoides aphids. The population dynamics of apterous aphids went through five consecutive stages: initial, fluctuating, rising, peak and declining. The population dynamics of alate aphids was 4-7 days later than that apterous aphid's. Significant positive correlations were found between the population size of spiders and ladybugs which were natural enemies and number of aphids. The result of pesticides against aphids in field trial showed that 25% thiamethoxam WG, 70% imidacloprid WG and 20% acetamiprid WP had well controlling effect., Conclusion: Aphids on L. macranthoides could be well controlled while 25% thiamethoxam WG, 70% imidacloprid WG and 20% acetamiprid WP are sprayed during the period of aphid population raising, the early April to the mid May.

Objective: To explore the feasibility of ultra-rapid freezing of human spermatozoa in the cryogenic vial with different concentrations of sucrose solution., Methods: We divided 40 normal semen samples prepared with the routine swim-up technique into 6 aliquots, 1 as the control and the other 5 cryopreserved with sucrose solution at the concentrations of 0.15, 0.20, 0.25 and 0.30 mol/L, respectively. After thawing, we determined and compared the motility, progressive motility and plasma membrane integrity of the sperm among the 6 groups., Results: The motility, progressive motility and plasma membrane integrity of the sperm were significantly lower after thawing than before cryopreservation ([96.2 +/- 1.8]%, [93.8 +/- 2.8]% and [99.0 +/- 0.8 ]%) (P<0.05). Post-thawing sperm motility was (55.5 +/- 6.3)% in the 0.20 mol/L sucrose group, significantly higher than in the 0.15, 0.25 and 0.30 mol/L groups ([45.9 +/- 6.6]%, [50.4 +/- 9.4]% and [45.5 +/- 11.2]%) (P<0.05), and it was (53.6 +/- 5.0)% in the conventional freezing group, with no statistically significant difference from the 0.20 and 0.25 mol/L sucrose cryopreservation groups (P> 0.05), but remarkably higher than in the 0.15 and 0.30 mol/L groups (P<0.05). Post-thawing progressive sperm motility exhibited no statistically significant differences between the 0.20 mol/L sucrose and conventional freezing groups ([44.4 +/- 7.4]% vs [42.3 +/- 8.1]%, P>0.05), but markedly higher in both than in the 0.15, 0.25 and 0.30 mol/L sucrose groups ([37.1 +/- 8.3 ]%, [33.1 +/- 9.2]% and [22.0 +/- 9.1]%) (P<0.05). Post-thawing plasma membrane integrity was significantly higher in the 0.20 mol/L sucrose cryopreservation group ( [70.1 +/- 6.9]%) than in either the conventional freezing group ([63.1 +/- 6.8]%) or the 0.15, 0.25 and 0.30 mol/L sucrose groups ([57.7 +/- 8.3]%, [63.5 +/- 10.7]% and [57.8 +/- 12.9]%) (P<0.05)., Conclusion: As a simple, safe and effective method, ultra-rapid freezing with sucrose solution at the final concentration of 0.20 mol/L can be used for the cryopreservation of human spermatozoa.

Background: Before human MII oocytes are vitrified they are usually denuded from their cumulus cells. In this study we wanted to investigate the effects of an intact corona radiata on the vitrification and fertilization of human oocytes., Methods: The study comprised two different parts. In Part 1, 36 MII stage oocytes, from 6 patients, were randomly assigned into a control group, a group of vitrified-warmed oocytes without a corona radiata and a group of vitrified-warmed oocytes with an intact corona radiata. In each group of 12, 6 oocytes were used for evaluation of the zona pellucida solubility (hardening) and another 6 oocytes were used for the analysis of their ultrastructure. In addition, six polyspermically fertilized oocytes were used as positive controls for zona pellucida hardening. In Part 2, 16 patients in total produced 107 fresh and 98 vitrified-warmed oocytes, with or without an intact corona radiata. All oocytes were fertilized via conventional IVF and embryos were transferred according to our standard ET routines. The oocyte survival and fertilization rates, embryo quality and pregnancy and implantation rates were evaluated., Results: There were no differences in oocyte survival, zona pellucida solubility (hardening) or the number of cortical granules between the vitrified-warmed and fresh oocytes. There were also no differences in the zona pellucida solubility and the number of cortical granules between vitrified-warmed oocytes with or without an intact corona radiata. However, the oocytes with an intact corona radiata had a higher fertilization rate after conventional IVF insemination. No differences were seen in the survival and cleavage rates, the percentage of high-quality embryos or the clinical outcome., Conclusions: Zona hardening and ultrastructural damage do not seem to occur in vitrified human oocytes. An intact corona radiata in vitrified-warmed oocytes retains their fertilization capacity in conventional IVF, but does not improve the embryo quality. Poor fertilizing capacities of vitrified-warmed oocytes without an intact corona radiata seem to have been due to the complete removal of the cumulus cells.

Chronic psychosocial stress negatively affects ovarian function. Ovarian follicular development is regulated by both pituitary-derived gonadotropins and intraovarian regulatory factors. To date, the suppressive effects of chronic stress on the ovary have been observed to be manifested mainly as an inhibition of gonadotropin release. It is not clear whether there are any other intraovarian regulatory mechanisms involved in this process. Growth and differentiation factor 9 (GDF9) is an important, oocyte-specific paracrine regulator required for follicular development. In this study, the chronic unpredictable mild stress model was used to produce psychosocial stress in mice. The number of different developmental stages of follicles was counted on ovarian sections stained with hematoxylin and eosin. Real-time PCR and Western blotting were used to detect the mRNA and protein levels, respectively, of GDF9. The results show that chronic unpredictable stress inhibits follicular development, increases follicular atresia, and suppresses GDF9 expression. Exogenous gonadotropin treatment partly restores the repressed antral follicular development, but has no effect on the repressed secondary follicular development associated with chronic stress. Treatment with recombinant GDF9 restores secondary follicular development. Cotreatments with GDF9 and gonadotropins restore both secondary and antral follicular development in stressed mice. These findings demonstrate that inhibition of follicular development induced by chronic unpredictable stress is associated with GDF9 and gonadotropin.

Background: Brain-derived neurotropic factor (BDNF) was originally described in the nervous system but has been shown to be expressed in ovary tissues recently, acting as a paracrine/autocrine regulator required for developments of follicles and oocytes. Although it is generally accepted that chronic stress impairs female reproduction and decreases the expression of BDNF in limbic structures of central nervous system, which contributes to mood disorder. However, it is not known whether chronic stress affects oocytes developments, nor whether it affects expression of BDNF in ovary., Methods: Mice were randomly assigned into control group, stressed group, BDNF-treated group and BDNF-treated stressed group. The chronic unpredictable mild stress model was used to produce psychosocial stress in mice, and the model was verified by open field test and hypothalamic-pituitary-adrenal (HPA) axis activity. The methods of immunohistochemistry and western blotting were used to detect BDNF protein level and distribution. The number of retrieved oocytes, oocyte maturation, embryo cleavage and the rates of blastocyst formation after parthenogenetic activation were evaluated., Results: Chronic unpredictable stress decreased the BDNF expression in antral follicles, but didn't affect the BDNF expression in primordial, primary and secondary follicles. Chronic unpredictable stress also decreased the number of retrieved oocytes and the rate of blastocyst formation, which was rescued by exogenous BDNF treatment., Conclusion: BDNF in mouse ovaries may be related to the decreased number of retrieved oocytes and impaired oocytes developmental potential induced by chronic unpredictable stress.

The aim of this study was to utilize self-assembled polycaprolactone (PCL)-grafted chondroitin sulfate (CS) as an anticancer drug carrier. We separately introduced double bonds to the hydrophobic PCL and the hydrophilic CS. The modified PCL was reacted with the modified CS through a radical reaction (CSMA-g-PCL). The copolymer without doxorubicin (DOX) was noncytotoxic in CRL-5802 and NCI-H358 cells at a concentration ranging from 5-1000 μg/mL and DOX-loaded CSMA-g-PCL (Micelle DOX) had the lowest inhibitory concentration of 50% cell growth values against the NCI-H358 cells among test samples. The cellular uptake of Micelle DOX into the cells was confirmed by flow cytometric data and confocal laser scanning microscopic images. The in vivo tumor-targeting efficacy of Micelle DOX was realized using an NCI-H358 xenograft nude mouse model. The mice administered with Micelle DOX showed suppressed growth of the NCI-H358 lung tumor compared with those administered with phosphate-buffered saline and free DOX.

Purpose: Lipoprotein-associated phospholipase A₂ (Lp-PLA₂) is an inflammatory enzyme expressed in atherosclerotic plaques. We investigated the association of circulating Lp-PLA₂ with characteristics of vulnerable coronary atherosclerotic plaques., Materials and Methods: We recruited 113 patients with either unstable angina (UA, n=59) and stable angina (SA, n=54) by coronary angiography. Thirty-six healthy subjects served as controls. Intravascular ultrasound (IVUS) was used to evaluate the characteristics of coronary atherosclerotic plaque, and serum Lp-PLA₂ concentration was measured as well., Results: Lp-PLA₂ concentration was significantly higher in both UA and SA patients [(396±36) μg/L and (321±39) μg/L, respectively] compared with the controls [(127 ± 49) μg/L, p<0.01], and higher in UA than SA group. IVUS findings showed that remodeling index (RI) (0.91 ± 0.15 vs. 0.85 ± 0.11, p=0.005) and eccentricity index (EI) (0.73 ± 0.16 vs. 0.65 ± 0.22, p=0.039) were larger in UA than in SA group, and fibrous caps were thicker in SA than UA group [(0.91 ± 0.23) mm vs. (0.63 ± 0.21) mm, p=0.032]. Moreover, Lp-PLA₂ correlated positively with EI (r=0.439, p<0.01) and RI (r=0.592, p<0.05) in UA group. There was an inverse relationship between Lp-PLA₂ and fibrous cap thickness in both UA (r=-0.587, p<0.001) and SA (r=-0.318, p<0.05) groups. The independent risk factors in UA group were Lp-PLA₂ (OR=1.055, 95% CI: 1.03-1.08, p=0.013), LDL-cholesterol (OR=0.032, 95% CI: 0.00-0.05, p=0.041) and fibrous cap thickness (OR=0.008, 95% CI: 0.00-0.45, p=0.019). Lp-PLA₂ was strongly associated with both EI and fibrous cap thickness in both groups., Conclusion: Serum level of Lp-PLA₂ is associated with both eccentricity index and fibrous cap thickness in both UA and SA groups. Elevated levels of circulating Lp-PLA₂ might to be a strong risk factor and more serious for unstable angina than stable angina.

Problem: What is the relationship between Th17 cells and unexplained recurrent spontaneous abortion (URSA)?, Method of Study: CD4(+) interleukin (IL)-17A(+) T (Th17) cells and CD4(+) CD25(bright) Foxp3(+) regulatory T (Treg) cells in peripheral blood were analyzed by flow cytometry; IL-17 concentrations in cell culture supernatants were quantitatively determined by enzyme-linked immunosorbent assay; and IL-17A positive cells in decidua tissues were measured by immunohistochemistry., Results: The proportion of Th17 cells and IL-17A concentrations was both significantly higher in patients with URSA than in normal early pregnant (NEP) and non-pregnant (NP) patients, Treg frequencies were significantly lower in patients with URSA than in NEP patients, and the ratio of Th17 to Treg was significantly higher in the URSA group than in the other two. Additionally, the percentage of IL-17A cells in deciduas was significantly higher in patients with URSA than in NEP patients., Conclusions: Human Th17 cells may play a major role in rejecting conceptus antigens and therefore may be harmful to the maintenance of pregnancy. The data also suggest that Treg cells are beneficial to pregnancy. There may exist a balance of Th17/Treg in normal pregnancy., (© 2010 John Wiley & Sons A/S.)

Objective: To establish a simple, rapid and easy method for screening the gene mutation in hemophilia A, which was further applied to a direct diagnosis and carrier detection at gene level., Methods: Twenty-four clinically diagnosed hemophilia pedigrees, including all the hemophilia patients and female members, were tested for the introns 22 and 1 in factor VIII gene by using inversion polymerase chain reaction (PCR) and regular PCR techniques. All the 26 exons of factor VIII gene were consecutively screened in the 17 patients manifesting non-inverted sequences in intron 22 by using PCR, subsequently all the 37 amplicons resulted from 26 exons were analyzed by conformation sensitive gel electrophoresis (CSGE), finally the mutated exons were subjected to sequencing verification. According to the mutation results, mothers and twin sisters of the hemophilia probands were tested by CSGE or subjected to nucleotide sequencing directly, to ascertain if those individuals had the same mutation or were the carriers of disease-causing gene., Results: Intron 22 inversion was detected in 7 hemophilia probands out of 24 hemophilia pedigrees, intron 1 inversion was not detected in these pedigrees. Single-base mutations distributed in different exons of factor VIII gene were detected in 13 pedigrees with family history and 3 sporadic pedigrees, diagnosed as non-inverted 22 intron patients. By comprehensive usage of PCR-CSGE and nucleotide sequencing, the positive rate and the diagnosable rate of gene diagnosis or carrier detection in the 24 hemophilia pedigrees was 94.12% and 100% respectively., Conclusion: PCR-CSGE is a highly sensitive and special assay for detecting single base mutation. By integrated utilization of introns 22 and 1 of factor VIII gene detection and PCR-CSGE genotyping, combining with nucleotide sequencing, a direct diagnosis of all hemophilia pedigrees be could nearly make at gene level, including the sporadic families. This method might be used to screen new mutation theoretically and ascertain the mutation type. It is a simple, rapid and low-cost method, possessing unique advantages in direct diagnosis of hemophilia A and carrier screening. It should have important application value in hemophilia diagnosis.

Objective: To explore the relation between changes in expression of aquaporin-4(AQP4) protein and acute radiation-induced cerebral edema after Gamma knife radiation., Methods: The experimental model was established in rats by radiating 2 mm right to median line in cerebral hemisphere with 50 Gy Gamma knife (the center located at the line of auditory canal). The changes of brain water content were measured by the wet and dry weight method. Immunohistochemistry was used to determine the change of expression of AQP4 protein at different periods after Gamma knife radiation., Results: The brain water content and the expression of AQP4 in the target area and the peripheral zone obviously increased at 12 h after Gamma knife radiation, and achieved the peak after 2 d. It was still higher than normal after 14 d. The gray value of expression of aquaporin-4 was negatively related to brain water content (r=-0.9857, P<0.05)., Conclusion: AQP4 has a close relationship with acute radiation-induced cerebral edema.

Aim: To construct the vector and express anti-HIV-1 envelope glycoprotein single chain Fv fragment in Pichia pastoris., Methods: The target gene was digested from plasmid pET28-scFv and cloned into pichia pastoris vector via gene engineering and DNA recombination techniques. The recombinant plasmid was linearized and transferred into Pichia pastoris strains GS115 by electroporation. After positive recombinant was selected and expression was induced by methanol, the target protein was analyzed by RT-PCR, SDS-PAGE and double-antibody sandwich ELISA., Results: High copies of transformant were obtained by phenotype determining and PCR amplification. RT-PCR and SDS-PAGE demonstrated the target protein was successfully expressed. And the yield account for about 18 percent of the total cell proteins. Double-antibody sandwich ELISA analysis proved that the recombinant scFv had good biological activities since it could be recognized and induce special immune respond with gp120 antigen., Conclusion: The scFv was expressed successfully. This research will lay the foundation for AIDS target therapy and further study of anti-HIV activities.

Objective: To investigate the proportion and function of CD(4)(+)CD(25)(+) regulatory T cells (CD(4)(+)CD(25)(+) Tr) in unexplained recurrent spontaneous abortion (URSA)., Methods: (1) Proportion measurement: the proportion of CD(4)(+)CD(25)(+) Tr cells in peripheral blood was measured by double-label flow cytometric analysis. The samples were taken from 15 URSA women, 15 normal non-pregnancy women and 13 normal pregnancy women. (2) Function measurement: CD(4)(+)CD(25)(+) Tr cells and CD(4)(+)CD(25)(-) T cells were extracted from peripheral blood lymphocytes by the microbeads separation. The purity of CD(4)(+)CD(25)(+) Tr cells and CD(4)(+)CD(25)(-) T cells was measured by flow cytometry. The growth inhibitory effect of CD(4)(+)CD(25)(+) Tr cells on CD(4)(+)CD(25)(-) T cells was assessed in vitro., Results: The proportion of CD(4)(+)CD(25)(+) Tr cells was decreased significantly in URSA women (6.9 +/- 1.8)% than that in normal non-pregnancy women [(10.8 +/- 1.1)%] (P<0.05) and normal pregnancy women [(11.2 +/- 1.4)%] (P<0.01). Moreover, the suppressive rate of CD(4)(+)CD(25)(+) Tr cells was decreased significantly in URSA women (75 +/- 6)% than that in normal non-pregnancy women [(89 +/- 4)%] (P<0.05) and normal pregnancy women [(90 +/- 4)%] (P<0.01). However, with respect to the proportion and function of CD(4)(+)CD(25)(+) Tr cells, there was no significant difference between normal non-pregnancy women and normal pregnancy women (P > 0.05)., Conclusion: The results suggest that decrease in proportion and function of CD(4)(+)CD(25)(+) Tr cells may be associated with URSA.

Objective: To investigate a new method of genioplasty for treatment of a prominent mandibular angle with mandibular malformation., Methods: According to the different types of the mandible, the prominent mandibular angle was grinded with the masseter muscle kept intact. Autologous micromorselized bone of the grinded mandibular angle was collected and implanted into the chin for augmentation mentoplasty, and the chin was fixed with gypsum after operation., Results: Fifteen patients receiving the surgery were followed up for 6 -12 months, and satisfactory effects were achieved in 12 patients. One patient requested modification of the shape of the chin 10 days after the operation, and another two sustained chin distortion due to external forces and underwent subsequent shape adjustment of the chin., Conclusion: The procedure described avoids graft rejection and causes less complications for genioplasty with easy moulding and satisfactory outcome.

Objective: To probe into the role of electroacupuncture in treatment of peripheral facial paralysis., Methods: Eighty-six cases were divided into a treatment group (n=45) treated with electroacupuncture plus Chinese medicine, and a control group (n=41) treated with acupuncture plus Chinese medicine. Taiyang (EX-HN 5), Yangbai (GB 14) and other points on the affected side were selected and Chinese medicine modified Qianzheng powder was administrated. Their clinical therapeutic effects were analyzed and evaluated., Results: The effective rate was 97. 8% in the treatment group and 82.9% in the control group with a significant difference between the two groups (P < 0.05). the treatment group being higher than the control group., Conclusion: Chinese medicine plus acupuncture combined with electroacupuncture has a better therapeutic effect on peripheral facial paralysis.