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From the perspective of biomechanics and forming technology, Ti−Fe−Zr−Sn−Y eutectic alloy was designed using a “cluster-plus-glue-atom” model, and then the alloy was prepared by laser additive manufacturing (LAM) on pure titanium substrate. The mechanical properties of the alloy were evaluated using micro-hardness and compression tester, and the elastic modulus was measured by nanoindenter. The results show that the alloy exhibits a high hardness of HV (788±10), a high strength of 2229 MPa, a failure strain of 14%, and a low elastic modulus of 87.5 GPa. The alloy also has good tribological, chemical, forming, and biological properties. The comprehensive performances of the Ti64.51Fe26.40Zr5.86Sn2.93Y0.30 alloy are superior to those of the Ti70.5Fe29.5 eutectic alloy and commercial Ti−6Al−4V alloy. All the above-mentioned qualities make the alloy a promising candidate as LAM biomaterial.

Rationale and Objectives The purpose of this study was to explore conventional MRI features that can accurately differentiate central nervous system embryonal tumor, not otherwise specified (CNS ETNOS) from glioblastoma (GBM) in adults. Materials and Methods Preoperative conventional MRI images of 30 CNS ETNOS and 98 GBMs were analyzed by neuroradiologists retrospectively to identify valuable MRI features. Five blinded neuroradiologists independently reviewed all these MRI images, and scored MRI features on a five-point scale. Kendall's coefficient of concordance was used to measure inter-rater agreement. Diagnostic value was assessed by the area under the curve (AUC) of receiver operating curve, and sensitivity and specificity were also calculated. Results Seven MRI features, including isointensity on T1WI, T2WI, and FLAIR, ill-defined margin, severe peritumoral edema, ring enhancement, and broad-based attachment sign, were helpful for the differential diagnosis of these two entities. Among these features, ring enhancement showed the highest inter-rater concordance (0.80). Ring enhancement showed the highest AUC value (0.79), followed by severe peritumoral edema (0.67). The combination of seven features showed the highest AUC value (0.86), followed by that of three features (ill-defined margin, severe peritumoral edema, and ring enhancement) (0.83). Conclusion Enhancement pattern, peritumoral edema, and margin are valuable for the discrimination between CNS ETNOS and GBM in adults.

Ovarian cancer is one of the most malignant gynecological tumors in the world, with a high fatality rate and resistance to chemotherapy. Much basic research has revealed that oxidative stress is involved in tumor occurrence and development processes and is associated with tumor progression as well as metastasis. In recent years, oxidative stress has been proven to have a close connection with malignant biological behavior in ovarian cancer, and this topic has garnered increasing attention. This article reviews the research aimed toward elucidating the relationship between oxidative stress and ovarian cancer.

The standard treatment for advanced ovarian cancer is cytoreductive surgery followed by cytotoxic chemotherapy. However, it has high risk of recurrence and poor prognosis. Poly(ADPribose) polymerase (PARP) inhibitors selectively target DNA double-strand breaks (DSBs) in tumor cells that cannot be repaired and induce the synthetic lethality of BRCA1/2 mutation cancers. PARP inhibitors are clinically used to treat recurrent ovarian cancer and show significant efficacy in ovarian cancer patients with homologous recombination repair (HRR) pathway defects. PARP inhibitors also have significant clinical benefits in patients without HR defects. With the increasingly extensive clinical application of PARP inhibitors, the possibility of acquiring drug resistance is high. Therefore, clinical strategies should be adopted to manage drug resistance of PARP inhibitors. This study aims to summarize the indications and toxicity of PARP inhibitors, the mechanism of action, targeted treatment of drug resistance, and potential methods to manage drug-resistant diseases. We used the term “ovarian cancer” and the names of each PARP inhibitor as keywords to search articles published in the Medical Subject Headings (MeSH) on Pubmed, along with the keywords “clinicaltrials.gov” and “google.com/patents” as well as “uspto.gov.” The FDA has approved olaparib, niraparib, and rucaparib for the treatment of recurrent epithelial ovarian cancer (EOC). Talazoparib and veliparib are currently in early trials and show promising clinical results. The mechanism underlying resistance to PARP inhibitors and the clinical strategies to overcome them remain unclear. Understanding the mechanism of resistance to PARP inhibitors and their relationship with platinum resistance may help with the development of antiresistance therapies and optimization of the sequence of drug application in the future clinical treatment of ovarian cancer.

With the rapid development of rail transit, storage of mass data has become an urgent problem. In view of the problem of insufficient storage capacity and low-efficiency for traditional storage technology, maintenance monitoring data collected by MSS (Maintenance Support System) is taken as the background in this paper [1], an open source framework Hadoop is introduced to build MSS data storage architecture. Through building the virtual cluster environment, the Hadoop technology used in signal system of rail transit is verified feasible.

Sapphire (α-Al2O3) single crystal combines many good mechanical and optical properties that make it become the material of choice in a variety of modern Hi-Tech applications The use of CMP technique can produce high quality surface finishes at low cost and with fast material removal rates. In CMP process, nanoabrasive is very important parameters in determining the removal rate. In this paper, the influence mechanism of nanoabrasive on sapphire substrate CMP was analyzed deeply. The main kinetics processing was discussed in detail. According to sapphire substrate properties, alkali slurry was chosen and the mix of SiO2 and Al2O3 sol was selected as nanoabrasive, and the particle size was 20~30nm. The results show that adding nanoalumina with the concentration of 20ml / L in the main abrasive can make the chemical action balance with the mechanical action in the CMP process, so that the substrate removal rate is well improved, and the surface roughness can be reduced to 0.236nm by adjusting the process parameters and slurry ratio. Such data meets the super smooth requirement and the optimal sapphire surface can be gotten. So such slurry with mixed abrasive can be as rough polishing slurry and can improve the polishing efficiency.

The Zn0.2Ca0.8TiO3:0.1Pr3+ luminescent materials were prepared by sol-gel technology. The phase transformation of gel was studied by TG-DTA and XRD in heat treatment process. The luminescent properties of phosphor in different calcination temperatures were determined by fluorescence spectrophotometer, and then the best sintering temperature was confirmed. The luminescent properties of phosphor was obtained through the emission/ excitation spectra. The surface appearance measured with SEM. The results showed that the nanoZn0.2Ca0.8TiO3:0.1Pr3+ phosphor could be obtained by sol-gel, sintering at 900°C. It sent out red fluorescent under the Ultra Violet. This characteristic emission peak at 614nm is associated with the typical 1D23H4 transition of Pr3+ion.

Although neuritic plaques and neurofibrillary tangles in older adults are correlated with cognitive impairment and severity of dementia, it has long been recognized that the relationship is imperfect, as some people exhibit normal cognition despite high levels of Alzheimer's disease (AD) pathology. We compared the cellular, synaptic, and biochemical composition of midfrontal cortices in female subjects from the Religious Orders Study who were stratified into three subgroups: (1) pathological AD with normal cognition ("AD-Resilient"), (2) pathological AD with AD-typical dementia ("AD-Dementia"), and (3) pathologically normal with normal cognition ("Normal Comparison"). The AD-Resilient group exhibited preserved densities of synaptophysin-labeled presynaptic terminals and synaptopodin-labeled dendritic spines compared with the AD-Dementia group, and increased densities of glial fibrillary acidic protein astrocytes compared with both the AD-Dementia and Normal Comparison groups. Further, in a discovery-type antibody microarray protein analysis, we identified a number of candidate protein abnormalities that were associated with a particular diagnostic group. These data characterize cellular and synaptic features and identify novel biochemical targets that may be associated with resilient cognitive brain aging in the setting of pathological AD.

Choriocarcinoma is one of the gestational trophoblastic neoplasias (GTNs) that originate in the chorionic villi and the extravillous trophoblast. Long noncoding RNAs (lncRNAs) are a type of non-protein-coding RNAs that have recently been implicated in human tumorigenesis. The present study investigated the role of the lncRNA LINC00261 in cell proliferation, metastasis, and apoptosis in choriocarcinoma cell lines. The transcription level of LINC00261 was significantly lower in choriocarcinoma tissues and in choriocarcinoma cell lines. Overexpression of LINC00261 caused a decrease in cell proliferation and arrested the cell cycle at the G0/G1 phase. Furthermore, overexpression of LINC00261 inhibited cell migration and invasion. Meanwhile, it promoted cell apoptosis and the relative activities of caspase 3 and caspase 9 in choriocarcinoma JEG-3 and JAR cells. These data suggested that LINC00261 promotes cell proliferation and metastasis in choriocarcinoma. Our data might provide novel insight into the early diagnosis and treatment of choriocarcinoma in clinics.

While a potential causal factor in Alzheimer's disease (AD), brain insulin resistance has not been demonstrated directly in that disorder. We provide such a demonstration here by showing that the hippocampal formation (HF) and, to a lesser degree, the cerebellar cortex in AD cases without diabetes exhibit markedly reduced responses to insulin signaling in the IR→IRS-1→PI3K signaling pathway with greatly reduced responses to IGF-1 in the IGF-1R→IRS-2→PI3K signaling pathway. Reduced insulin responses were maximal at the level of IRS-1 and were consistently associated with basal elevations in IRS-1 phosphorylated at serine 616 (IRS-1 pS⁶¹⁶) and IRS-1 pS⁶³⁶/⁶³⁹. In the HF, these candidate biomarkers of brain insulin resistance increased commonly and progressively from normal cases to mild cognitively impaired cases to AD cases regardless of diabetes or APOE e4 status. Levels of IRS-1 pS⁶¹⁶ and IRS-1 pS⁶³⁶/⁶³⁹ and their activated kinases correlated positively with those of oligomeric Aβ plaques and were negatively associated with episodic and working memory, even after adjusting for Aβ plaques, neurofibrillary tangles, and APOE e4. Brain insulin resistance thus appears to be an early and common feature of AD, a phenomenon accompanied by IGF-1 resistance and closely associated with IRS-1 dysfunction potentially triggered by Aβ oligomers and yet promoting cognitive decline independent of classic AD pathology.

Animal models provide compelling evidence that chronic stress is associated with biochemical and morphological changes in the brain, many of which are mediated by corticosterone, a principal glucocorticoid synthesized in the rodent adrenal cortex and secreted in response to stress. To better characterize the effects of chronic corticosterone at the synaptic and sub-synaptic level, we implanted 3-month-old male C57B/6 mice with 2 × 5 mg corticosterone pellets (CORT group, n=14), 21 day release formulation (20 mg/kg/day dose) or placebo pellets (Placebo group, n=14), 21 day release formulation. After 20 days, brains were removed. One hemisphere was frozen for biochemical analysis by synaptosomal fractionation with Western blotting, and the other hemisphere was fixed for immunohistochemistry. Localization and expression levels for PSD-95, NR1, and synaptopodin proteins were assessed. Biochemical analysis revealed lower protein levels of PSD-95 (32% decrease, p

Ni-WC-MgO compound coatings were made by way of Vacuum Fusion Sintering (VFS). Effects of MgO addition level on the microstructures of coating surface and longitudinal section, element diffusion in fusion sintering area and wear resistance were analyzed through scanning electron microscope (SEM) and energy dispersive spectrometer (EDS). The results showed as the following: MgO addition made coating and matrix combined closely; there existed obvious transition region. When the addition level of MgO was 0.5%, the coating had the fewest defects and best combination. Ni, Cr, W, C and other elements in the coating diffused to the direction of the base metal while Fe in the base metal diffused to the coating, during which the diffusion of Fe and Ni was most obvious at the junction. Therefore, when the addition level of MgO was 0.5%, the surface microstructure of the coating was the densest and finest, and the wear resistance of the coating was the best.

Olfactory dysfunction is an early and common sign in various neurodegenerative diseases. Microsmia or anosmia is present in approximately 90% of patients with Alzheimer’s disease (AD)1 and accumulating evidence suggests that this psychophysical biomarker predicts incident mild cognitive impairment (MCI),2 conversion of MCI to AD,3 more rapid cognitive decline in older adults initially free of dementia,4 and also correlates with severity of dementia and abundance of neurodegenerative disease pathology in the brain.5, 6 Olfactory deficits are found in other neurodegenerative disorders as well, especially Parkinson’s disease7, 8 and Huntington disease,9 and to varying degrees in frontotemporal dementias and corticobasal syndromes,10 pure autonomic failure,11 multiple system atrophy,12 Guamanian ALS-Parkinson’s-Dementia Complex,13 idiopathic ALS,14 progressive supranuclear palsy,15 and cerebellar degenerations.16, 17 The neuropathological basis of olfactory dysfunction in AD and PD is thought to be due mainly to accumulations of disease-related lesions, especially tau pathology in AD andα-synuclein pathology in PD, that occur in the olfactory bulb and primary olfactory sensory cortices of the cerebrum.18–23 The olfactory epithelium (OE) is a pseudostratified columnar epithelium lying deep within the recesses of the superior nasal cavity (for review, see24). It is composed of a mixture of mulitpotential and committed stem cells (basal cells), supporting cells, and chemosensory olfactory receptor neurons. Mature neurons give rise to fine, unmyelinated axons that form bundles (olfactory fila) that ascend through the cribriform plate to synapse in the olfactory bulb. Early research on the OE in AD described tau and neurofilament immunoreactive dystrophic neurites in the lamina propria adjacent to the epithelium layer in subjects with AD but not control subjects.25, 26 However, subsequent studies reported that dystrophic neurites, which also were found to express ubiquitin, α-, β-, and γ-synucleins, and amyloid-β, could be found in normal aging and other neurodegenerative diseases as well.27–30 In light of this presumed non-specificity, interest in the OE for diagnostic or mechanistic research into AD, PD, and other neurodegenerative diseases waned. Still, these previous studies are relatively few in number, had small sample sizes in each diagnostic group, and may have been otherwise confounded by the limited sensitivity and specificity of reagents available at the time and a variety of other technical challenges inherent in working with OE tissues. Thus, the frequency, extent, and disease specificity of these lesions and other forms of pathological protein expression have not been established, and the degree to which they may be associated with brain pathology or olfactory dysfunction is not known. Over the last twenty years, we have collected OE in many neurodegenerative disease and non-neurological control cases that have been autopsied in protocols of the University of Pennsylvania’s Center for Neurodegenerative Disease Research (CNDR). At the same time, there have been steady improvements in the sensitivity and specificity of immunochemical reagents to reveal pathological proteins in tissue, as well as the discovery of new proteins, such as TDP-43, that play important roles in neurodegenerative diseases.31 Here we present findings in a large sample of neuropathologically verified AD and control cases, as well as other neurodegenerative diseases representing a spectrum of tauopathies, synucleinopathies, and TDP-43 proteinopathies.

Progressive synaptic degeneration and neuron loss are major structural correlates of cognitive impairment in Alzheimer's disease (AD). The mechanisms by which synaptic degeneration in AD occurs have not been established. The activation of proteins within the caspase family has been implicated in AD-associated neurodegeneration, and synaptically localized caspase activity could play a role in the synaptic degeneration and loss found in AD. We used synaptosomal fractionation with Western blotting and immunohistochemistry to examine the anatomical, subcellular, and subsynaptic expression patterns of caspase 3 in both the anterior cingulate cortex and hippocampus of control and AD patients. In both control and AD cases, there was a selective enrichment of caspase- 3 at synapses, particularly in the postsynaptic density (PSD) fractions. Compared with controls, AD patients exhibited significant increases in synaptic procaspase- 3 and active caspase-3 expression levels that were most evident in the PSD fractions. These data demonstrate for the first time the preferential localization and increase of caspase-3 in the PSD fractions in AD and suggest an important role for caspase 3 in synapse degeneration during disease progression.

Early cancer metastases often occur in cervical cancer (CC) patients, resulting in poor prognosis and poor therapeutic outcome after resection of primary cancer. Hence, there is a compelling requirement for elucidating the molecular mechanisms underlying the CC cell invasiveness. Recently, the role of microRNAs (miRNAs) and pituitary tumor-transforming gene 1 (Pttg1) in the carcinogenesis of CC has been reported. Nevertheless, the relationship between miRNAs and Pttg1 remains ill-defined. Here, we showed that the levels of miR-3666 were significantly decreased and the levels of zinc finger E-box binding homeobox 1 (ZEB1) and Pttg1 were significantly increased in the CC specimens from patients, compared to the paired non-tumor tissue. Moreover, the levels of miR-3666 and ZEB1 inversely correlated. Bioinformatics analyses showed that miR-3666 targeted the 3′-untranslated region (3′-UTR) of ZEB1 messenger RNA (mRNA) to inhibit its translation, which was confirmed by luciferase reporter assay. Moreover, Pttg1 overexpression inhibited miR-3666 and subsequently increased ZEB1 and cell invasion, while Pttg1 depletion increased miR-3666 and subsequently decreased ZEB1 and cell invasion. Together, our data suggest that Pttg1 may increase CC cell metastasis, possibly through miR-3666-regulated ZEB1 levels.

The birth and differentiation of neurons have been extensively studied in the olfactory epithelium (OE) of rodents but not in humans. The goal of this study was to characterize cellular composition and molecular expression of human OE in vivo and in vitro. In rodent OE, there are horizontal basal cells and globose basal cells that are morphologically and functionally distinct. In human OE, however, there appears to be no morphological distinction among basal cells, with almost all cells having round cell bodies similar to rodent globose basal cells. Unlike the case in rodents, human basal cells, including putative neuronal precursors, express p75NGFR, suggesting a distinctive role for p75NGFR in human OE neurogenesis. Molecular expression of neuronal cells during differentiation in human OE grossly follows that in rodents. However, the topographical organization of immature and mature ORNs in human OE differs from that of rodents, in that immature and mature ORNs in humans are dispersed throughout the OE, whereas rodent counterparts have a highly laminar organization. These observations together suggest that the birth and differentiation of neuronal cells in human OE differ from those in rodents. In OE explant culture, neuronal cells derived from human OE biopsy express markers for immature and mature neurons, grossly recapitulating neuronal differentiation of olfactory neurons in vivo. Furthermore, small numbers of cells are doubly label for bromodeoxyuridine and olfactory marker protein, indicating that neuronal cells born in vitro reach maturity. These data highlight species-related differences in OE development and demonstrate the utility of explant culture for experimental studies of human neuronal development.

Frontotemporal degeneration (FTD) is a neurodegenerative condition that has been principally associated with frontal lobe dementia. In this study, we compared neuropathological abnormalities in frontal, hippocampal, and calcarine cortices from patients assigned a diagnosis of FTD, normal elderly and Alzheimer's disease (AD). Densities of Nissl-stained neurons and lesions which were immunolabeled for tau, beta-amyloid (Abeta), alpha- and beta-synuclein, ubiquitin, glial fibrillary acidic protein (GFAP) and CD68 antigen were determined using computer-assisted, non-biased quantitative microscopy. We found that FTD frontal and hippocampal regions exhibited marked neuron loss, abundant ubiquitin-immunoreactive (ir) dystrophic neurites, GFAP-ir astrocytes, and CD68-ir microglia, while calcarine cortex was spared. No alpha- or beta-synuclein-ir lesions were observed, and neither the density of tau-ir neurofibrillary tangles nor that of Abeta-ir plaques in FTD exceeded normal controls. In addition, there were no neuropathological differences between FTD subjects who presented clinically with a frontal lobe dementia versus an AD-like dementia. These findings indicate that FTD is a category of neurodegnerative dementias with varying clinical presentations that is characterized by the progressive degeneration of select populations of cortical neurons. The molecular neurodegenerative mechanisms that lead to FTD remain to be elucidated.

Neuronal insulin signaling abnormalities have been associated with Alzheimer's disease (AD). However, the specificity of this association and its underlying mechanisms have been unclear. This study investigated the expression of abnormal serine phosphorylation of insulin receptor substrate 1 (IRS1) in 157 human brain autopsy cases that included AD, tauopathies, α-synucleinopathies, TDP-43 proteinopathies, and normal aging. IRS1-pS(616), IRS1-pS(312) and downstream target Akt-pS(473) measures were most elevated in AD but were also significantly increased in the tauopathies: Pick's disease, corticobasal degeneration and progressive supranuclear palsy. Double immunofluorescence labeling showed frequent co-expression of IRS1-pS(616) with pathologic tau in neurons and dystrophic neurites. To further investigate an association between tau and abnormal serine phosphorylation of IRS1, we examined the presence of abnormal IRS1-pS(616) expression in pathological tau-expressing transgenic mice and demonstrated that abnormal IRS1-pS(616) frequently co-localizes in tangle-bearing neurons. Conversely, we observed increased levels of hyperphosphorylated tau in the high-fat diet-fed mouse, a model of insulin resistance. These results provide confirmation and specificity that abnormal phosphorylation of IRS1 is a pathological feature of AD and other tauopathies, and provide support for an association between insulin resistance and abnormal tau as well as amyloid-β.

The cellular distribution of inositol 1,4,5-trisphosphate receptors was examined in rodent maxillary incisor teeth. In situ hybridization studies with a transmembrane probe of type I inositol 1,4,5-trisphosphate receptor indicated that this receptor/channel was highly expressed in odontoblast cells of incisor teeth. In contrast, very low labeling was observed in dental pulp. Northern analysis showed a message size of approximately 9.5 kilobases for this receptor, and demonstrated that type III inositol 1,4,5-trisphosphate receptor was expressed in incisor teeth. Immunocytochemical studies confirmed that types I and III inositol 1,4,5-trisphosphate receptors were both highly expressed in odontoblasts while very low expression was detected in dental pulp. Finally, antibodies that recognized alpha subunits of the Gq class of GTP binding proteins also stained odontoblasts. These results indicate that receptor-mediated regulation of calcium release through inositol 1,4,5-trisphosphate receptors may occur in odontoblasts of rat incisor teeth. These findings also suggest that inositol 1,4,5-trisphosphate receptor/channels regulate calcium flux in odontoblasts during mineralization of dentin, or in growth and differentiation of incisor tissue.

Aim and Background: Cervical cancer remains the third most common cancer in women globally after breast and colorectal cancer. Well-characterized biomarkers are necessary for early diagnosis and to predict metastatic progression and effective therapy. MiRNAs can regulate gene expression, cell growth, differentiation and apoptosis by targeting mRNAs for translational repression or degradation in tumor cells. The present study was conducted to assess expression of miR93, miR200a, RECK, MMP2, MMP9 in invasive cervical carcinoma, and analyze their clinical significance. Method: A total of 116 patients with invasive cervical carcinoma and 100 patients undergoing hysterectomy for benign lesions were retrospectively examined. Quantitative real-time PCR was performed to determine expression of miR93 and miR200a while RECK, MMP2, MMP9 and MVD were assessed by immunohistochemical staining. Results: Cervical carcinoma patients demonstrated up-regulation of miR-93, miR-200a, MMP2 and MMP9, with down-regulation of RECK as compared to benign lesion tissues. RECK was significantly inversely related to invasion and lymphatic metastasis. The 5-year survival rate for patients with strong RECK expression was significantly higher than that with weakly expressing tumors. Conclusion: MiR-93 and miR-200a are associated with metastasis and invasion of cervical carcinoma. Thus together with RECK they are potential prognostic markers for cervical carcinoma. RECK cooperating with MMP2, MMP9 expression is a significant prognostic factor correlated with long-term survival for patients with invasive cervical carcinoma.

To explore the alteration of collagen ultrastructure and content in uterine ligaments and paraurethral tissue and explore whether the alteration may contribute to stress urinary incontinence (SUI) and pelvic organ prolapse (POP).The cardinal ligament, uterosacral ligament and paraurethral tissue samples were obtained from 90 subjects undergoing hysterectomy. Collagen ultrastructure was examined with transmission electron microscopy. And collagen content and expression of vasoactive intestinal peptide (VIP) were examined with immunohistochemistry.The smooth muscle fascicles were thinner in the patients of SUI and POP. Arrangement of smooth muscle fascicles was disorderly. Fibroblast was metabolically active. The mean collagen fibril diameters in the SUI and POP groups were larger than that in the control group (P0.01). The mean contents of collagen I and III in the SUI and POP groups were lower than that in the control group (P0.01). The expression of VIP was lower (P0.05).Predominance of collagen degradation during tissue repair may contribute to and promote POP and SUI. The decrease of VIP might be related with nerve damage or degeneration to cause or accelerate the progress of pelvic organ prolapse.

To explore the feasibility and safety of mdr1 gene transferred into placenta derived mesenchymal stem cells (P-MSCs) by reconstructed retroviral vector.Human P-MSCs were isolated and expanded by Percoll density gradient and then transduced repeatedly by reconstructed retroviral vector containing mdr1 gene. The transfection and expression of mdr1 gene was detected by reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS). Meanwhile, the biological features of mdr1-MSCs were identified and analyzed.The expression of mdr1mRNA was found in transfected cells. The expression of P-glycoprotein (P-gp) encoded by mdr1 gene was (27.6 ± 5.1)% in the transfected P-MSCs cells versus (0.4 ± 0.1)% in the non-transfected P-MSCs cells (t = 14.291, P0.01). The percent of P-MSCs at quiescent phase (G0/G1 phase) was around 95.40% and it was in accord with the characterization of stem cells. The mdr1-MSCs exhibited typical ultrastructures of low-differentiated stem cells. Moreover, they still retained the potency of adipogenic and osteogenic differentiation in the presence of appropriate conditioned media.A stable expression of P-gp may be obtained by reconstructed retroviral-mediated transfection in vitro. And transfected MSCs retain the characteristics of stem cells.

Between 3 and 25 months of age, light and electron microscopic features of principal neurons in the central nucleus of the inferior colliculus of the C57BL/6 mouse were quantitated. This mouse strain has a genetic defect producing progressive sensorineural hearing loss which starts during young adulthood (2 months of age) with high-frequency sounds. During the second year of life, hearing is severely impaired, progressively involving all frequencies. The hearing loss was documented in the present study by auditory brainstem recordings of the mice at various ages. The cochleas from many of the same animals showed massive loss of both inner and outer hair cells beginning at the base (high-frequency region) and progressing with age along the entire length to the apex (low-frequency region). In the inferior colliculi, there was a significant decrease in the size of principal neurons in the central nucleus. There was a dramatic decrease in the number of synapses of all morphologic types on principal neuronal somas. The percentage of somatic membrane covered by synapses decreased by 67%. A ventral (high frequency) to dorsal (low frequency) gradient of synaptic loss could not be identified within the central nucleus. These synaptic changes may be related to the equally dramatic physiologic changes which have been noted in the central nucleus of the inferior colliculus, in which response properties of neurons normally sensitive to high-frequency sounds become more sensitive to low-frequency sounds. The synaptic loss noted in this study may be due to more than the loss of primary afferent pathways. It may represent alterations of the complex synaptic circuitry related to the central deficits of presbycusis.

Myelosuppression is the main dose-related toxicity of many chemotherapeutic drugs. The human multidrug resistance (mdr1) gene is well-known for its ability to confering drug resistance. In this study, we meant to transplant the placenta mesenchymal stem cells (P-MSCs) moderated by mdr1 gene into a nude mice model radiated by γ-Co(60) and to explore the chemoprotection for bone marrow (BM) toxicity.Human P-MSCs were isolated from trypsin-digested term placentas and then transduced by with reconstructed retroviral vector containing mdr1 gene and green fluorescent protein (GFP) reporter gene. The integration and expression of mdr1 gene was observed indirectedly by the expression of GFP. A nude mice model was constructed after irradiation with a sublethal dosage of γ-Co(60). These irradiated mice were transplanted with mdr1-MSCs through the caudal vein and then received paclitaxel (PAC) intraperitoneal chemotherapy. The Peripheral peripheral blood (PB) of the nude mice was collected, and the PB cells counts and values were determined using an automatic analyzer.After PAC treatment, mdr1-MSCs transplanted mice showed markedly improved survival upon compared to MSCs transplanted mice (85.7% vs. 57.1%). White blood cell (WBC) and red blood cell (RBC) counts as well as the hemoglobin (Hb) values were significantly increased in PAC treated mdr1-MSCs mice compared to PAC treated control mice when PAC chemotherapy had been finished (all P0.05), but the difference was not found in the plateltes (PLT) count (P0.05).Human P-MSCs moderated by mdr1 gene when transplanted into nude mice may provide chemoprotection for hematopoietic toxicity.

To transfer multidrug resistance gene (mdr1) into human placental mesenchymal stem cells (P-MSCs) by retroviral vector and assess the effects of mdr1 gene transduction upon biological features of P-MSCs.Human P-MSCs were isolated from trypsin-digested term placentas and then transduced by reconstructed retroviral vector containing mdr1 gene and green fluorescent protein (GFP) reporter gene. Flow cytometric analysis was employed to determine the immunophenotypes of transfected P-MSCs. And the proliferation and cell cycle were detected by methyl thiazolyl tetrazolium and propidium iodide staining. Ultrastructures of transfected P-MSCs were observed and different induction conditions used to direct the cells to differentiate into adipocytes and osteoblasts.The transfected P-MSCs still expressed stem cell markers such as CD29, CD44 and CD73. The mean cumulative time of population doubling was 23.9 hours. The cellular cycle retained the proliferative characterization of stem cells. Ultrastructural features of transfected P-MSCs included increased surface microvilli, abundant mitochondria and slightly swollen rough endoplasmic reticulum. Furthermore these transfected cells demonstrated osteogenic and adipogenic differentiation potentials under appropriate conditions.The mdr1 gene transduction by retroviral vector in vitro has no significant effect upon biological characteristics of P-MSCs. It might provide experimental references for the application of P-MSCs in high-dose tumor chemotherapy.

To investigate the effects of ovarian carcinoma cells on the differentiation, maturation, and function of the dendritic cells (DC).Human epithelial ovarian carcinoma cells were isolated from specimens of ovarian carcinoma from 12 patients obtained during operation, and cultured. Peripheral blood samples were collected from these patients. DC were isolated and co-cultured with recombinant human tumor necrosis factor (TNF)-alpha to promote their maturation. Immunostaining and flow cytometry (FC) were used to detect the DC specific marker CD1a, maturation marker CD83, co-stimulating factors CD86 and human leucocyte antigen (HLA)-DR. FITC-labeled glucan was added and FC was used to detect the phagocytic function. T cells were co-cultured with immature and mature DC of different concentrations respectively for 96 h, 18 h before the end of culture 3H-TdR was added, liquid scintillation counter was used to measure the level of count per minute (cpm) Ovarian carcinoma cells and peripheral mononuclear cells were put into Transwell to contact each other directly or indirectly, TNF-alpha was add to promote the maturation, and then FC was used to detect the CD1a, CD83, CD86, and HLA-DR levels, and the glucan uptake level. ELISA was used to detect the levels of IL-12, IL-10, and interferon (IFN)-gamma, and Western blotting was used to detect the p-38 protein and phosphorylated p-38 protein in the supernatant.After stimulation of TNF-alpha the expression; levels of CD1a, CD86, CD83, and HLA-DR of the DCs were remarkably increased. Immature DCs showed a strong ability at phagocytosis of glucan, and this ability was decreased by the stimulation of TNF-alpha. The proliferative function of mature DCs on allogenic T lymphocytes was significantly increased dose-dependently (P0.05). Co-culture with ovarian carcinoma cells decreased the expression levels of CD1a, CD86, and HLA-DR, increased the expression level of CD83, and decreased the ability in up taking glucan by 43.05%. The proliferative function of the DC cultured in direct contact with ovarian carcinoma cells was decreased by 56.35%, and such inhibitory function of the DCs cultured not in direct contact with ovarian carcinoma cells was weaker, and the levels of CD1a, CD86, CD83, and HLA-DR expressed by the DC cultured not in direct contact with ovarian carcinoma cells was between those of the DC cultured in direct contact with ovarian carcinoma cells and those of the DC undergoing routine culture. The levels of IL-12 and phosphorylated p-38 protein were decreased in the supernatant of the DC co-cultured with ovarian carcinoma cells.Ovarian carcinoma cells suppress the differentiation of monocyte into DC, negatively regulates the phagocytotic and antaean managing functions of DC, thus the DC can not effectively stimulate the proliferation of T lymphocytes and inhibit the activity of cytotoxic T lymphocytes. That may be one of the underlying mechanisms of immune escape of tumor and immune tolerance of T lymphocytes.

To develop a new method to promote the differentiation of mesenchymal stem cells derived from human placenta (pMSC) to uterus smooth muscle cells (uSMC) in simulated uterus microenvironment.MSCs were isolated from human placenta, cultivated, and analyzed for their phenotype by flow cytometry. The multipotential differentiation of the pMSC was examined by chondrogenic, adipogenic, and osteogenetic induction. uSMC were isolated from uteri resected during operation and co-cultivated with the pMSC in a Transwell chamber simulating Two, 4, and 8 days later RT-PCR and Western blotting were used to detect the mRNA and protein expression of alpha-actin, calmodulin, and myosin heavy chains (MHC), the markers of smooth muscle differentiation at the early, middle, and late stages. On day 8 RT-PCR was used to detect the expression of estrogen receptor in these 2 groups of cells, then estrogen was used to stimulate these cells and the protein kinase C (PKC) activity was examined.The pMSC could be induced into adipocytes, osteocytes, and chondrocytes respectively. After co-culture with uSMC, the morphology of the pMSC changed closely into that of the uSMC, and MHC was expressed in the pMSC. Estrogen receptor was positive in both groups of cells. The PKC activity increased, especially in the cell membrane, after stimulation of estrogen.The postpartum human placenta can be used as an important and novel source of multipotent stem cells for tissue engineering and genetic engineering. Placental MSC have the potential to differentiate into smooth muscle cells under the simulated uterus microenvironment in vitro.

BACKGROUND Most of gynecologic malignancies are sensitive to chemotherapy. Myelosuppression is the main dose-related toxicity of many chemotherapeutic drugs. The human multidrug resistance (mdr1) gene is well known for its ability to confer drug resistance. This study aimed to explore the feasibility of expression and resistance of mdr1 gene transduction into human placenta mesenchymal stem cells (P-MSCs) by retrovirus vector. METHODS Human P-MSCs were isolated from trypsin-digested term placentas, and their immunophenotypes and differentiation potential were evaluated. Human P-MSCs were transduced by reconstructed retroviral vector containing the mdr1 gene and green fluorescent protein (GFP) reporter gene. The integration and expression of the mdr1 gene were observed indirectly by the expression of GFP, and fluorescence-activated cell sorter was used to evaluate the functional activity of permeability glycoprotein (P-gp) encoded by the mdr1 gene. The stimulating test was made in vitro to show pleiotropic drug resistance of transfected cells. RESULTS The isolated, cultured and expanded P-MSCs expressed stem cell markers such as CD29, CD44 and CD73, and showed osteogenic and adipogenic differentiation potentials under appropriate conditions. The expression of P-gp in the non-transfected P-MSCs cells was (0.4 +/- 0.1)%, but increased to (28.1 +/- 4.7)% after gene transfection (P < 0.01). And positive staining of P-gp located mainly at cell membrane and cytoplasm. Accumulation and extrusion assays showed that P-gp expressed by the transfected cells had pump-functional activity and could efflux daunomycin out of cells. The analysis of cell survival confirmed that transfected P-MSCs had a characteristic of multidrug resistance with a significant increase in the resistance to anticancer agents. CONCLUSIONS Transfer and expression of human mdr1 gene mediated by retrovirus vector conferred P-MSCs drug resistance. It might provide a new alternative to chemoprotection strategies.

To investigate the effect of dendritic cells (DCs) pulsed by oligonucleotide containing "un-methylated cytimidine-phosphodiester bond-guanylic acid" motif (CpG ODN) on ovarian carcinoma cells.Dendritic cells were isolated form the peripheral monocytes and co-incubated with synthesized CpG2006, CAI125, important epithelial ovarian cancer-associated antigen, or CpG ODN + CA125 for 72 h. Flow cytometry (FC) was used to detect the expression of CD(1alpha), CD(63), CD(86), and human leucocyte antigen-DR (HLA-DR) on the cell surface. ELISA was used to detect the IL-12 level in the supernatant. T cells were obtained from the DCs. The suspensions of different groups of pulsed DCs were co-incubated with T cells. Mixed lymphocyte reaction (MLR) was used to detect the proliferating activity of the T cells. Ovarian carcinoma cells of the line OVCAR-3 were added into the culture fluid of T cells with different effector-target ratios, and then unpulsed DCs, CpG ODN-pulsed DCs, CpG ODN + CA125-pulsed DCs, and CA125-pulsed DCs were added respectively. MTT colorimetry was performed to measure the A values of different wells so as to calculate the killing rate.(1) The expression levels of CD(63), CD(86), and HLA-DR on the membranes of the CpG ODN-pulsed DCs and CpG ODN + C125-pulsed DCs were significantly higher than those of the un-pulsed DCs (all P0.01), however, there were no significant differences in the expression of CD(1alpha) among different groups (all P0.05). (2) The IL-12 levels in the supernatants of the CpG ODN-pulsed DCs and CpG ODN + C125-pulsed DCs were significantly higher than those of the unpulsed and CA125-pulsed groups (all P0.01). (3) MLR showed that the T cell proliferation rates of the T cells sensitized by the CpG ODN-pulsed DCs and CpG ODN + C125-pulsed DCs were both higher than those of the T cells stimulated by the unpulsed and CA125-pulsed groups when the effector: target ratio was 10:1 (all P0.01). (4) The killing rates on OVCAR-3 cells of the CTLs sensitized by CpG ODN + CA125-pulsed DCs at the same effector: target ratios were all higher than those of the CTLs sensitized by the CpG ODN-pulsed DCs, CA125-pulsed DCs, and unpulsed DCs (all P0.01), the killing activity of the cytotoxic T lymphocytes (CTLs) sensitized by CpG ODN+CA125-pulsed DCs on the OVCAR-3 cells was shown even when the effector: target ratio was as low as 10:1, and then increased along with the increase of the effector: target ratio to the height of 64.9%.Capable of inducing immune cytotoxicity on ovarian carcinoma, DCs sensitized by CpG ODN + CA125 may have a great implication on clinical application.

Variations in the gene encoding the novel protein dysbindin-1 (DTNBP1) are among the most commonly reported genetic variations associated with schizophrenia. Recent studies show that those variations are also associated with cognitive functioning in carriers with and without psychiatric diagnoses, suggesting a general role for dysbindin-1 in cognition. Such a role could stem from the protein's known ability to affect neuronal glutamate release. How dysbindin-1 might affect glutamate release nevertheless remains unknown without the discovery of the protein's neuronal binding partners and its subcellular locus of action. We demonstrate here that snapin is a binding partner of dysbindin-1 in vitro and in the brain. Tissue fractionation of whole mouse brains and human hippocampal formations revealed that both dysbindin-1 and snapin are concentrated in tissue enriched in synaptic vesicle membranes and less commonly in postsynaptic densities. It is not detected in presynaptic tissue fractions lacking synaptic vesicles. Consistent with that finding, immunoelectron microscopy showed that dysbindin-1 is located in (i) synaptic vesicles of axospinous terminals in the dentate gyrus inner molecular layer and CA1 stratum radiatum and in (ii) postsynaptic densities and microtubules of dentate hilus neurons and CA1 pyramidal cells. The labeled synapses are often asymmetric with thick postsynaptic densities suggestive of glutamatergic synapses, which are likely to be derived from dentate mossy cells and CA3 pyramidal cells. The function of dysbindin-1 in presynaptic, postsynaptic and microtubule locations may all be related to known functions of snapin.

The FOXO family of forkhead transcription factors stimulates the transcription of target genes involved in many fundamental cell processes, including cell survival, cell cycle progression, DNA repair, and insulin sensitivity. The activity of FOXO proteins is principally regulated by activation of protein kinase B (PKB)/Akt by insulin and other cytokines. PKB/Akt phosphorylates three consensus sites in FOXO proteins, leading to their export from the nucleus and the inhibition of FOXO-stimulated transcription. It has been widely accepted that the decreased transcription results from reduced abundance of FOXO proteins in the nucleus. In the present study we mutated Leu 375 to alanine in the nuclear export signal of Foxo1 (mouse FOXO1), so that it would remain in the nucleus of H4IIE rat hepatoma cells after insulin treatment, and determined whether insulin could still inhibit transcription stimulated by the Foxo1 mutant. Despite the retention of the Foxo1 mutant in the nucleus, insulin inhibited L375A-Foxo1-stimulated transcription to the same extent as transcription stimulated by wild-type Foxo1. Similar results were obtained using reporter plasmids containing the rat IGF-binding protein-1 promoter or a minimal promoter with three copies of the insulin response element to which FOXO proteins bind. We conclude that insulin can inhibit Foxo1-stimulated transcription even when nuclear export of Foxo1 is prevented, indicating that insulin inhibition can occur by direct mechanisms that do not depend on altering the subcellular distribution of the transcription factor. (Endocrinology 144: 5615–5622, 2003)

Abnormally phosphorylated tau accumulates as neurofibrillary tangles and neuropil threads in older persons with and without Alzheimer's disease. The relationship between neurofibrillary tangles and neuropil threads and how they relate to cognitive function is unknown. This study investigated the relationship between phosphorylated tau lesions and cognitive function in 31 persons participating in the Religious Orders Study, a prospective, longitudinal clinicopathological study of aging and Alzheimer's disease. All subjects underwent detailed neuropsychological performance testing within a year of death and evidenced a spectrum of cognitive performance ranging from normal abilities to mild dementia. Measures of neurofibrillary tangle density and phosphorylated tau immunoreactive structures (predominantly neuropil threads) in the entorhinal and perirhinal cortices by quantitative image analysis were significantly correlated (r = 0.5). In multiple linear regression analyses controlling for age, sex, and education, parahippocampal neurofibrillary tangles and neuropil threads were significantly lower in persons without cognitive impairment compared to those with mild cognitive impairment and/or Alzheimer's disease. Further, neurofibrillary tangles were significantly correlated to measures of episodic memory but not other cognitive abilities; neuropil tangles were not significantly related to memory or other cognitive functions. These data indicate that phosphorylated tau pathology in the ventromedial temporal lobe develop prior to the onset of clinical dementia and their presence is associated with cognitive impairment, particularly impairment of episodic memory.

In the title salt, [Cd(C6H15NO3)2](C8H4O4), the Cd2+cation is coordinated by six O atoms and two N atoms from two tetradentate 2-[bis(2-hydroxyethyl)amino]ethanol ligands, displaying a distorted square-antiprismatic coordination. The terephthalate dianion does not coordinate to the cation but is connected through O...H—O hydrogen bonds of medium strength to the complex cations, leading to a layered structure extending parallel to (100).

Pathological alterations in dendrites and axons (i.e., neuritic pathologies) occur in the normal aging brain as well as in brains from elders with mild cognitive impairment and neurodegenerative dementia. These alterations may correlate with clinical measures of cognitive abilities, but the contribution of neuropil threads (NTs), which constitute 85–90% of cortical tau pathology, has not been clear because of the lack of quantitative methodologies. We combined quantitative fractionation and image analysis to devise a strategy for measuring the burden of tau-rich NTs in the entorhinal and perirhinal cortex of brains from elders with and without cognitive impairment, including dementia due to Alzheimer's disease (AD). On the basis of data presented here using this novel strategy, we conclude that this quantitative imaging technique will facilitate efforts to determine the behavioral correlations of neuritic lesions in AD and other brain disorders.