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Estrogen and progestins have adverse effects, and many of these adverse effects are caused by progestins. Due to this, many women choose to use botanical alternatives for hormone replacement therapy, which does not trigger steroidogenic properties. Therefore, it is necessary to screen these herbs for progestogenic and anti-progestogenic properties. Extract of 13 Chinese medicinal plants were analysed for progestogenic and anti-progestogenic activities by using progesterone response element-driven luciferase reporter gene bioassay. MTT assay was carried out to investigate the cytotoxic effect of herb extract on PAE cells. Among the 13 herbs, Dipsacus asperoides extract exhibited progestogenic activity, and 10 species - Cortex eucommiae, Folium artemisiae argyi, Glycyrrhiza uralensis, Angelica sinensis, Atractylodes macrocephala koidz, Scutellaria baicalensis, Cuscuta chinensis, Euscaphis japonica, Ailanthus altissima, and Dioscorea opposita - were recognized to have anti-progestogenic like activities. Extract of Dipsacus asperoides demonstrated dose-dependent progestogenic activity, and the progestogenic activity of 100 (mu)g/mL extracts was equivalent to 31.45 ng/mL progesterone activity. Herbs extracts that exhibited anti-progestogenic-like activity also inhibited the 314.46 ng/mL progesterone activity in a dose-response manner. None of the herb extracts shown significant toxic effect on PAE cells at 40-100 (mu)g/mL compared to control. This discovery will aid selection of suitable herbs for hormone replacement therapy.

Structural optimization of the prior lead 3 led to the small molecular (D)-leucinamides with potent modulating activity and Notch-sparing selectivity on the proteolytic processing of amyloid-β precursor proteins. The N-(R)-epoxypropyl analog 10c exhibited potent γ-secretase modulation compared to DAPT and showed substantial substrate selection for APP cleavage over Notch cleavage, while N-(2-fluoro)benzyl analog 10e showed the most potent γ-secretase inhibition with dull selectivity. The exceptional suppression of ERK-mediated activation suggested that these potent γ-secretase modulators may adapt an alternative pathway to prominently induce the differential inhibition of C99 cleavage by γ-secretase., (Copyright © 2014 Elsevier Masson SAS. All rights reserved.)

The present study was undertaken to evaluate the hepatoprotective effect mechanisms of Nelumbo nucifera leaves extract (NLE) in experimental alcoholic steatohepatitis animal models. We found that the NLE contained polyphenols (phenolic acids and flavonoids), and more than 70% of the main functional components in NLE could potentially provide benefits for alcoholic liver disease. The parameters of histopathology, immunohistochemistry, antioxidant defense, proinflammatory mediator and lipid synthesis-related proteins demonstrated the inhibitory effect of NLE on alcoholic steatohepatitis. Plasma and hepatic content analysis showed that NLE inhibited lipid accumulation by altering the levels of triglycerides (TG) and cholesterol (TC). Treatment with NLE increased the expression of the p-AMPK/AMPK ratio and PPAR-α. Furthermore, fatty acid oxidation and transport via carnitine palmitoyltransferase-1 (CPT1) and microsomal triglyceride transfer protein (MTP) were through the activation of the AMPK and PPAR-α signal. These results revealed that the polyphenol-rich component of NLE prevents alcoholic steatohepatitis by multiple pathways, including reduced lipid synthesis, enhanced fatty acid oxidation and transport responses, inhibited oxidative stress and facilitated anti-inflammation. Suggesting that NLE might be regarded as a beneficial food that has the potential to be developed as a natural agent for preventing alcoholic steatohepatitis.

Purpose: To obtain the thin coronal sectional anatomic dataset of the liver by using digital freezing milling technique., Methods: The upper abdomen of one Chinese adult cadaver was selected as the specimen. After CT and MRI examinations verification of absent liver lesions, the specimen was embedded with gelatin in stand erect position and frozen under profound hypothermia, and the specimen was then serially sectioned from anterior to posterior layer by layer with digital milling machine in the freezing chamber. The sequential images were captured by means of a digital camera and the dataset was imported to imaging workstation., Results: The thin serial section of the liver added up to 699 layers with each layer being 0.2 mm in thickness. The shape, location, structure, intrahepatic vessels and adjacent structures of the liver was displayed clearly on each layer of the coronal sectional slice. CT and MR images through the body were obtained at 1.0 and 3.0 mm intervals, respectively., Conclusion: The methodology reported here is an adaptation of the milling methods previously described, which is a new data acquisition method for sectional anatomy. The thin coronal sectional anatomic dataset of the liver obtained by this technique is of high precision and good quality.

Objective: The aim of the current study was to compare the clinical outcomes of mesh fixation with fibrin sealant (FS) spray or mechanical stapling (MS) in laparoscopic total extraperitoneal hernioplasty (TEP)., Background: The most appropriate method of mesh fixation is uncertain., Methods: Between June 2007 and June 2011, consecutive patients with primary reducible unilateral inguinal hernia who underwent day-case laparoscopic TEP were recruited. Outcome parameters included the incidence of acute and chronic pain, recurrence rates, morbidity rates, analgesic requirements, quality-of-life (QOL) scores, and direct cost., Results: During the study period, 130 patients were included in the study. Patients in the MS group had significantly worse pain scores on the day after operation (P = 0.006). Analgesic requirements were similar between the 2 groups (P = 0.558). At 6 months, no significant differences in the incidence of chronic pain were observed (at rest, after coughing or cycling). The incidence of seroma formation was similar between the 2 groups (P = 0.64), and no recurrences were observed at 1 year. No differences in the QOL scores were detected. The direct cost of the entire hospitalization in the FS group was less expensive (P < 0.001)., Conclusions: FS and MS are both effective methods of providing mesh fixation. FS was associated with reduced acute pain but not chronic pain. The rates of seroma formation were similar. However, the use of FS for mesh fixation was less expensive. [corrected].

Resistin and endothelin (ET)-1 have been reported to inhibit adipogenesis and regulate adipocyte insulin resistance, respectively. Although both hormones interact with each other, the exact signaling pathway of ET-1 to act on resistin gene expression is still unknown. Using 3T3-L1 adipocytes, we investigated the signaling pathways involved in ET-1-stimulated resistin gene expression. The up-regulation of resistin mRNA expression by ET-1 depends on concentration and timing. The concentration of ET-1 that increased resistin mRNA levels by 100%-250% was approximately 100 nM for a range of 0.25-12 hours of treatment. Treatment with actinomycin D blocked ET-1-increased resistin mRNA levels, suggesting that the effect of ET-1 requires new mRNA synthesis. Treatment with an inhibitor of the ET type-A receptor, such as N-[1-Formyl-N-[N-[(hexahydro-1H-azepin-1-yl)carbonyl]-L-leucyl]-D-tryptophyl]-D-tryptophan (BQ610), but not with the ET type-B receptor antagonist N-[(cis-2,6-Dimethyl-1-piperidinyl)carbonyl]-4-methyl-L-leucyl-1-(methoxycarbonyl)-D-tryptophyl-D-norleucine (BQ788), blocked ET-1, increased the levels of resistin mRNA, and phosphorylated levels of downstream signaling molecules, such as ERK1/2, c-Jun N-terminal kinases (JNKs), protein kinase B (AKT), and signal transducer and activator of transcription 3 (STAT3). Moreover, pretreatment of specific inhibitors of either ERK1/2 (1,4-diamino-2,3-dicyano-1,4-bis[2-aminophenylthio]butadiene [U0126] and 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one [PD98059], two inhibitors of MEK1), JNKs (SP600125), phosphatidylinositol 3-kinase/AKT (LY294002 and Wortmannin), or Janus kinase 2 (JAK2)/STAT3 ((E)-2-Cyano-3-(3,4-dihydrophenyl)-N-(phenylmethyl)-2-propenamide, AG490) prevented ET-1-increased levels of resistin mRNA and reduced the ET-1-stimulated phosphorylation of ERK1/2, JNKs, AKT, and STAT3, respectively. However, the p38 kinase antagonist 4-[5-(4-Fluorophenyl)-2-[4-(methylsulfonyl)phenyl]-1H-imidazol-4-yl]pyridine (SB203580) did not alter the effect of ET-1. These results imply that ET type-A receptor, ERK1/2, JNKs, AKT, and JAK2, but not ET type-B receptor or p38, are necessary for the ET-1 stimulation of resistin gene expression. In vivo observations that ET-1 increased resistin mRNA and protein levels in sc and epididymal adipose tissues support the in vitro findings.

Objective: To compare the long-term anorectal manometry of patients with grade III hemorrhoids treated by procedure for prolapse and hemorrhoids (PPH) or Doppler-guided hemorrhoidal artery ligation (DGHAL)., Methods: A total of 114 patients with grade III hemorrhoids in our department in 2010 were prospectively enrolled, and divided into PPH group (63 cases) and DGHAL group (51 cases) according to patient preference. Patients received anorectal pressure examination 2, 6, 12, 18 months after operation, including rectal resting pressure, anal resting pressure, maximal anal contractive pressure and length of high-pressure band. Anorectal manometries between two procedure groups were compared., Results: In every time point of follow-up, as compared to PPH group, DGHAL group presented higher anal resting pressure[postoperative 2 months: (56.2±2.6) mmHg vs. (43.2±3.1) mmHg, postoperative 6 months: (55.2±2.1) mmHg vs. (45.1±2.5) mmHg, postoperative 12 months: (53.2±3.0) mmHg vs. (40.2±3.2) mmHg, postoperative 18 months: (55.3±2.6) mmHg vs. (43.2±2.5) mmHg, all P<0.01], and longer length of high-pressure band [postoperative 2 months: (3.45±0.14) cm vs. (2.95±0.22) cm, postoperative 6 months: (3.46±0.12) cm vs. (2.94±0.26) cm, postoperative 12 months: (3.45±0.12) cm vs. (3.01±0.21) cm, postoperative 18 months: (3.56±0.12) cm vs. (3.02±0.23) cm, all P<0.01]. While no significant differences of maximal anal contractive pressure and rectal resting pressure were found between the two groups., Conclusion: Both PPH and DGHAL are safe procedures and have similar efficacy in the treatment of grade III hemorrhoids, while DGHALpossesses better efficacy in the time of anal function recovery and the maintenance of anal function.

The ribosome is centrally situated to sense metabolic states, but whether its activity, in turn, coherently rewires transcriptional responses is unknown. Here, through integrated chemical-genetic analyses, we found that a dominant transcriptional effect of blocking protein translation in cancer cells was inactivation of heat shock factor 1 (HSF1), a multifaceted transcriptional regulator of the heat-shock response and many other cellular processes essential for anabolic metabolism, cellular proliferation, and tumorigenesis. These analyses linked translational flux to the regulation of HSF1 transcriptional activity and to the modulation of energy metabolism. Targeting this link with translation initiation inhibitors such as rocaglates deprived cancer cells of their energy and chaperone armamentarium and selectively impaired the proliferation of both malignant and premalignant cells with early-stage oncogenic lesions.

Background and Objective: Extramammary Paget's disease (EMPD) is a rare intra-epithelial neoplasm. Surgical excision is the treatment of choice. The usefulness of topical photodynamic therapy (PDT) in the treatment of EMPD has been reported. The objective of this prospective pilot study was to investigate the feasibility of combined PDT and surgery in the treatment of EMPD., Subjects and Methods: A total of 13 patients with 19 large EMPD lesions were recruited and assigned to surgery (n = 5) or PDT + surgery (n = 8) group. For the PDT + surgery group, four sessions of topical PDT mediated with 20% 5-aminolevulinic acid (ALA-PDT) were applied prior to surgery. Patients were followed up for 12 months. Treatment outcomes, adverse reactions and recurrence were compared., Results: In the surgery group, recurrence was seen in 2 out of 8 lesions (25%). In the combination group, over 58% reduction in lesion size was achieved after 4-sessions of PDT and recurrence was seen in 1 out of 11 lesions (9.1%) after surgery., Conclusions: Multiple ALA-PDT could be applied to reduce the severity of EMPD lesion and improve the success of surgery., (Copyright © 2013 Wiley Periodicals, Inc.)

Changes in DNA copy number, whether confined to specific genes or affecting whole chromosomes, have been identified as causes of diseases and developmental abnormalities and as sources of adaptive potential. Here, we discuss the costs and benefits of DNA copy-number alterations. Changes in DNA copy number are largely detrimental. Amplifications or deletions of specific genes can elicit discrete defects. Large-scale changes in DNA copy number can also cause detrimental phenotypes that are due to the cumulative effects of copy-number alterations of many genes simultaneously. On the other hand, studies in microorganisms show that DNA copy-number alterations can be beneficial, increasing survival under selective pressure. As DNA copy-number alterations underlie many human diseases, we will end with a discussion of gene copy-number changes as therapeutic targets., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Endothelin (ET)-1 and suppressor of cytokine signaling (SOCS)-3 were respectively found to regulate energy metabolism and hormone signaling in fat cells. Although ET-1 can also regulate the expression of SOCS-3-stimulating hormones, it is still unknown whether ET-1 regulates SOCS-3 gene expression. This study investigated the pathways involved in ET-1's modulation of SOCS-3 gene expression in 3T3-L1 adipocytes. ET-1 upregulated SOCS-3 mRNA and protein expression in dose- and time-dependent manners. The concentration of ET-1 that increased SOCS-3 mRNA levels by 250-400% was ∼100nM with 2-4h of treatment. Treatment with actinomycin D prevented ET-1-stimulated SOCS-3 mRNA expression, suggesting that the effect of ET-1 requires new mRNA synthesis. Pretreatment with the ET type A receptor (ET(A)R) antagonist, BQ-610, but not the ET type B receptor (ET(B)R) antagonist, BQ-788, prevented the stimulatory effect of ET-1 on SOCS-3 gene expression. The specific inhibitors of either MEK1 (U-0126 and PD-98059), JAK (AG-490), JNK (SP-600125), or PI3K (LY-294002 and wortmannin) reduced ET-1-increased levels of SOCS-3 mRNA and respectively inhibited ET-1-stimulated activities of MEK1, JAK, JNK, and PI3K. These results imply that the ET(A)R, ERK, JAK, JNK, and PI3K are functionally necessary for ET-1's stimulation of SOCS-3 gene expression. Moreover, ET-1 was observed to upregulate expressions of SOCS-1, -2, -3, -4, -5, and -6 mRNAs, but not SOCS-7 or cytokine-inducible SH2-containing protein-1 mRNAs. This suggests that ET-1 selectively affects particular types of SOCS family members. Changes in SOCS gene expressions induced by ET-1 may help explain the mechanism by which ET-1 modulates hormone signaling of adipocytes., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Through arranging of ancient books and literatures related pressing moxibustion, the history along with origin and development of pressing moxibustion is systematically investigated and analyzed, which provided theoretical basis for the clinical practice of pressing moxibustion. It is found that after 600 years of development and innovation, pressing moxibustion already has complete theoretical system which is reflected in the unitarity of formulating prescription, the diversity of the manipulation the universality of indications. What's more, the functional characteristics and mechanism of pressing moxibustion are initially discussed and its present research status and prospect in the field of modern moxibustion are revealed.

Background: While there is strong evidence for phosphatidylinositol 3-kinase (PI3K) involvement in cancer development, there is limited information about the role of PI3K regulatory subunits. PIK3R3, the gene that encodes the PI3K regulatory subunit p55γ, is over-expressed in glioblastoma and ovarian cancers, but its expression in gastric cancer (GC) is not known. We thus used genetic and bioinformatic approaches to examine PIK3R3 expression and function in GC, the second leading cause of cancer mortality world-wide and highly prevalent among Asians., Methods: Primary GC and matched non-neoplastic mucosa tissue specimens from a unique Asian patient gastric cancer library were comprehensively profiled with platforms that measured genome-wide mRNA expression, DNA copy number variation, and DNA methylation status. Function of PIK3R3 was predicted by IPA pathway analysis of co-regulated genes with PIK3R3, and further investigated by siRNA knockdown studies. Cell proliferation was estimated by crystal violet dye elution and BrdU incorporation assay. Cell cycle distribution was analysed by FACS., Results: PIK3R3 was significantly up-regulated in GC specimens (n = 126, p < 0.05), and 9.5 to 15% tumors showed more than 2 fold increase compare to the paired mucosa tissues. IPA pathway analysis showed that PIK3R3 promoted cellular growth and proliferation. Knockdown of PIK3R3 decreased the growth of GC cells, induced G0/G1 cell cycle arrest, decreased retinoblastoma protein (Rb) phosphorylation, cyclin D1, and PCNA expression., Conclusion: Using a combination of genetic, bioinformatic, and molecular biological approaches, we showed that PIK3R3 was up-regulated in GC and promoted cell cycle progression and proliferation; and thus may be a potential new therapeutic target for GC.

The scandium(III)-catalyzed enantioselective Hosomi-Sakurai allylation of isatins with various substituted allylic silanes is described. A catalyst loading as low as 0.05 mol % is utilized at room temperature to afford the 3-allyl-3-hydroxy-2-oxindoles in excellent yields and enantioselectivity up to 99% ee, including a demonstration of a gram-scale reaction. The effects of additives and varying silyl groups were explored to demonstrate the scope and application.

Objective: To study the molecular mechanism of apoptosis of leukemic cells (K562 cells) induced by iron chelating agent deferoxamine (DFO)., Methods: The exponentially growing K562 cells were used (1×10(6)/mL) in this study. The K562 cells were treated with different concentrations of DFO (10, 50 and 100 mmol/L), DFO+FeCl3 (10 μmol/L each) or normal saline (blank control). The cellular labile iron pool was measured with a fluorimetric assay using the metalsensitive probe calcein-AM. The viable count and cell viability were determined by typanblue assay. Cell apoptosis was determined by morphological study and flow cytometry assay. Caspase-3 activity in K562 cells was detected by colorimetry., Results: After DFO treatment, the cellular labile iron pool and the viability of K562 cells were reduced and the cell apoptosis increased in a time- and dose-dependent manner compared with the blank control group. The apoptosis rate of K562 cells in the DFO+FeCl3 treatment group was not significantly different from that in the blank control group. The caspase-3 activity in K562 cells increased significantly 24 hrs after 50 and 100 μmmol DFO treatment when compared with the blank control group (P<0.01). There was a negative correlation between cellular labile iron pool and caspase-3 activity of K562 cells (r=-0.894, P<0.05)., Conclusions: DFO induces apoptosis of leukemic cells possibly through decreasing cellular labile iron pool and increasing caspase-3 activity of the cells.

Aneuploidy, an incorrect chromosome number, is a hallmark of cancer. Compounds that cause lethality in aneuploid, but not euploid, cells could therefore provide new cancer therapies. We have identified the energy stress-inducing agent AICAR, the protein folding inhibitor 17-AAG, and the autophagy inhibitor chloroquine as exhibiting this property. AICAR induces p53-mediated apoptosis in primary mouse embryonic fibroblasts (MEFs) trisomic for chromosome 1, 13, 16, or 19. AICAR and 17-AAG, especially when combined, also show efficacy against aneuploid human cancer cell lines. Our results suggest that compounds that interfere with pathways that are essential for the survival of aneuploid cells could serve as a new treatment strategy against a broad spectrum of human tumors., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Aim: To study the expressional changes of soluble CD40 ligand(sCD40L) in peripheral blood serum and nuclear factor-κB (NF-κB) p65 in peripheral blood mononuclear cells (PBMC) of patients with acute progressive cerebral infarction(APCI)., Methods: We selected ninety-nine patients getting APCI less than 7 d of the onset as APCI group by prospective method. Each 100 cases of patients with acute cerebral infarction(ACI) in the same and with cerebral arteriosclerosis(CAS) in the outpatient were respectively selected as ACI and CAS group. The expressional changes of sCD40L in peripheral blood serum and NF-κBp65 in PBMC of patients with CAS on admission, of patients with APCI and ACI when in hospital, on the course of seventh day, of fourteen and of thirtieth were detected respectively., Results: The expression of sCD40L in peripheral blood serum and NF-κBp65 in PBMC of patients of ACI group on admission were obviously higher all than that of CAS group (P<0.05); The expression of sCD40L in peripheral blood serum and NF-κBp65 in PBMC of patients of APCI group when in hospital, on the course of seventh day, of fourteen and of thirtieth were obviously higher all than that of ACI group (P<0.05 )., Conclusion: The inflammatory and apoptotic mechanism mediated by expressional excessive increase of CD40-CD40L signal passage and NF-κB in PBMC might be one of the molecular biology mechanisms of onset and progress for APCI.

The purpose of this study was to explore the anatomical complexity and adjacent relationships of the sellar region in thin continuous sections so as to provide intimate morphological data for imaging diagnosis and surgical operations of the diseases in this region. After CT and MR examination verifying no brain lesions, one normal cadaver head was selected for this study from four Chinese adult male cadavers. After being embedded and frozen, the head was sliced into serial sections at 0.1 mm intervals in the transverse plane with SKC 500 computerized freezing milling machine. Then the serial transverse sections were photographed by a high-resolution digital camera and saved in the computer. Subsequently, the anatomic structures of the sellar region on the thin transverse sections were investigated and correlated with the MR images of the specimen as well as in vivo MR images, which were obtained from 20 normal Chinese male adult volunteers by a 3.0 T GE MR scanner. The base lines of the sectioning and the MR scan were all parallel to the AC-PC line. A total of 320 transverse sections and 10-12 transverse MR images related with the sellar region were obtained, respectively. We investigated the sectional anatomy of the sellar region and divided it into three parts: supra hypophysial area, hypophysial area and infra hypophysial area. The cavernous sinus was a venous passage full of blood and it could be divided into four interspaces according to its position relation with the internal carotid artery. The third, fourth, sixth cranial nerves and trigeminal branches ophthalmic nerve, maxillary nerve displayed from the anterior to the posterior in the lateral wall of cavernous sinus in transverse planes. Comparing continuous thin sections with MR images offers a better understanding of the complex anatomical structures and provides practical submillimeter anatomical data for imaging diagnosis and clinical treatment in this region.

The interaction between prulifloxacin (PUFX) and human serum albumin (HSA) was investigated under simulated physiologic conditions with fluorescence spectra. The fluorescence quenching process of HSA may be mainly governed by a static quenching mechanism. The apparent binding constant K(b) between PUFX and HSA at different temperatures were 2.08+/-1.04, 2.74+/-0.50, and 4.98+/-1.61x10(8) l/mol. The thermodynamic parameters, with a negative value of DeltaG(0), revealed that the binding is a spontaneous process. A binding distance R of 1.19 nm between donor and acceptor was obtained from the Fŏrster energy transfer theory.

Aims: To investigate the subcellular localization of Ca(v)2.2 calcium channel in the mouse central nervous system (CNS), and changes of Ca(v)2.2 at acute and chronic stages during and after pilocarpine-induced status epilepticus (PISE), in order to find out the roles it may play in epileptogenesis., Methods: Combined immunocytochemistry at both light and electron microscopic levels with real-time reverse transcription polymerase chain reaction (RT-PCR), cell transfection approach were used in this study., Results: N-type calcium channel Ca(v)2.2 subunit was distributed in different regions of the mouse CNS. It was mainly localized in the nuclei in different types of neurones and in astrocytes. At acute stages during and after PISE, Ca(v)2.2 expression decreased in the stratum pyramidale of CA3 area and in the stratum granulosum of the dentate gyrus, but increased in the stratum lucidum of CA3 area and in the hilus of the dentate gyrus. At chronic stage at 2 months after PISE, increased expression of Ca(v)2.2 in both the strata granulosum and molecular of the dentate gyrus was observed., Conclusions: Ca(v)2.2 is a nuclear protein in neurones and astrocytes in the mouse CNS. Its translocation occurs at acute stages during and after PISE. The increased expression of Ca(v)2.2 in both the strata granulosum and moleculare of the dentate gyrus at chronic stage at 2 months after PISE may be involved in the occurrence of spontaneously recurrent seizures.

Background: Stroke is the second most common cause of death in developed countries and a major cause of adult disability and mortality worldwide. New data strongly suggest that neuropeptide Y (NPY) may be a candidate gene for ischemic stroke., Methods: We investigated 450 ischemic stroke patients and 423 healthy controls matched for sex and age in a Han Chinese population. Three functional polymorphisms (-883TGins/del, -602G/T and -399 T/C) located in NPY gene promoter were genotyped using DNA sequencing methods., Results: Of 3 NPY polymorphisms investigated in our study, the -399CC genotype (OR: 1.699, 95% CI: 1.124-2.567, P=0.011) and the -399C allele (OR: 1.254, 95% CI: 1.031-1.524, P=0.023) were more frequent among ischemic stroke patients than in controls, especially in the small vessel disease (SVD) subtype patients. The similar results were observed in multivariable logistic regression analysis. Haplotype analysis revealed that the -883ins/-399C haplotype was a risk marker for ischemic stroke (P=0.008)., Conclusions: The C allele of -399 T/C polymorphism in the promoter regions of NPY is an independent risk factor for ischemic stroke, suggesting that NYP system may involve in the mechanisms of stroke pathology., (Copyright 2009 Elsevier B.V. All rights reserved.)

The goal of this study was to examine the morpho-physiologic changes in the dorsal subiculum network in the mouse model of temporal lobe epilepsy using extracellular recording, juxtacellular and immunofluorescence double labeling, and anterograde tracing methods. A significant loss of total dorsal subicular neurons, particularly calbindin, parvalbumin (PV) and immunopositive interneurons, was found at 2 months after pilocarpine-induced status epilepticus (SE). However, the sprouting of axons from lateral entorhinal cortex (LEnt) was observed to contact with surviving subicular neurons. These neurons had two predominant discharge patterns: bursting and fast irregular discharges. The bursting neurons were mainly pyramidal cells, and their dendritic spine density and bursting discharge rates were increased significantly in SE mice compared with the control group. Fast irregular discharge neurons were PV-immunopositive interneurons and had less dendritic spines in SE mice when compared with the control mice. When LEnt was stimulated, bursting and fast irregular discharge neurons had much shorter latency and stronger excitatory response in SE mice compared with the control group. Our results illustrate that morpho-physiologic changes in the dorsal subiculum could be part of a multilevel pathologic network that occurs simultaneously in many brain areas to contribute to the generation of epileptiform activity.

Aneuploidy refers to karyotypic abnormalities characterized by gain or loss of individual chromosomes. This condition is associated with disease and death in all organisms in which it has been studied. We have characterized the effects of aneuploidy on yeast and primary mouse cells and found it to be detrimental at the cellular level. Furthermore, we find that aneuploid cells exhibit phenotypes consistent with increased energy need and proteotoxic stress. These observations, together with the finding that the additional chromosomes found in aneuploid cells are active, lead us to propose that aneuploidy causes an increased burden on protein synthesis and protein quality-control pathways and so induces an aneuploidy stress response.

Multiple investigators have reported the presence of defects in the immune response of the elderly [Castle In: Clin Infect Dis 31:578, 2000; Ortqvist et al. In: Eur Respir J 30:414-422, 2007; Saurwein-Teissl et al. In: J Immunol 168:5893, 2002; Haynes et al. In: Proc Natl Acad Sci USA 100:15053-15058, 2003]. These defects reduce the magnitude of the immune response to infection and to vaccination. In individuals greater than 55 years of age, the probability of developing a fully protective neutralizing antibody response to the yearly multivalent particle inactivated influenza vaccine is less than 20% [Jefferson et al. In: Lancet 264:1165-1174, 2005; Goodwin et al. In: Vaccine 24:1159-1169, 2006; Jackson et al. In: Lancet 372:398-405, 2008; Simonsen and Taylor In: Lancet 7:658-666, 2007]. The defects in the aged immune system that are responsible for this limited response to vaccination in the older age groups include functional defects of the antigen presenting cells, functional defects in CD4 helper CD4 T cells and monocytes, and an altered microenvironment [Eaton et al. In: J Exp Med 200:1613-1622, 2004; Dong et al. In: J Gen Virol 84:1623-1628, 2003; Deng et al. In: Immunology 172:3437-3446, 2004; Cella et al. In: J Exp Med 184:747-752, 1996]. Starting at puberty, the involution of the thymus and the consequent reduction of the export of naïve T cells specific to neo-antigens leads to the reduction of the ratio of antigen naïve to memory cells as chronological age advances [Prelog In: Autoimmun Rev 5:136-139, 2006; McElhaney et al. In: J Immunology 176:6333-6339, 2006]. Changes in glycosylation of T cells and target antigens acquired during the aging process and the antibodies to these new glycopeptides and glycoproteins may also contribute to a reduction in the functioning of the adaptive immune response [Ishii et al. In: J Clin Neurosci 14:110-115, 2007; Shirai et al. In: Clin Exp Immunol 12:455-464, 1972; Adkins and Riley In: Mech Ageing Dev 103:147-164, 1998; Ben-Yehuda and Weksler In: Cancer Investigation 10:525-531, 1992]. One of the more interesting examples of the functional defects in the cells of the adaptive immune response is a reduced level of expression in the surface cytoadhesion and activation receptor molecules on CD4 helper T cells undergoing activation during vaccination. Upon infection or vaccination, CD40L is typically increased on the surface of CD4 helper T cells during activation, and this increased expression is absolutely essential to the CD40L promotion of expansion of antigen-specific B cells and CD 8 effector T cells in response to infection or vaccination [Singh et al. In: Protein Sci 7:1124-1135, 1998; Grewal and Flavell In: Immunol Res 16: 59-70, 1997; Kornbluth In: J Hematother Stem Cell Res 11:787-801, 2002; Garcia de Vinuesa et al. In: Eur J Immunol 29:3216-3224, 1999]. In aged human beings and mice, the reduced levels of expression of CD40 ligand (CD40L) in activated CD4 helper T cells is dramatically reduced [Eaton et al. In: J Exp Med 200:1613-1622, 2004; Dong et al. In: J Gen Virol 84:1623-1628, 2003]. To circumvent the reduction in CD40L expression and the subsequent reduction in immune response in the elderly, we have developed a chimeric vaccine comprised of the CD40L linked to the target antigen, in a replication incompetent adenoviral vector and in booster protein. This review will discuss the implementation the potential use of this approach for the vaccination of the older populations for cancer and infection.

Three-dimensional (3D) reconstruction of intrahepatic vessels is very useful in visualizing the complex anatomy of hepatic veins and intrahepatic portal vein. It also provides a 3D anatomic basis for diagnostic imaging and surgical operation on the liver. In the present study, we built a 3D digitized model of hepatic veins and intrahepatic portal vein based on the coronal sectional anatomic dataset of the liver. The dataset was obtained using the digital freezing milling technique. The pre-reconstructed structures were identified and extracted, and then were segmented by the method of manual intervention. The digitized model of hepatic veins and intrahepatic portal vein was established using 3D medical visualization software. This model facilitated a continuous and dynamic displaying of the hepatic veins and intrahepatic portal vein at different orientations, which demonstrated the complicated relationship of adjacent hepatic veins and intrahepatic portal vein realistically in the 3D space. This study indicated that high-quality 2D images, precise data segmentation, and suitable 3D reconstruction methods ensured the reality and accuracy of the digital visualized model of hepatic veins and intrahepatic portal vein.

Thrombolytic therapy is an important treatment for thrombosis, a life-threatening condition in cardiovascular diseases. However, the traditional thrombolytic therapies have often been associated with the risk of severe bleeding. By conjugating urokinase with magnetic nanoparticles (MNPs), we have presented a strategy to control thrombolysis within a specific site. The covalent bioconjugate of urokinase and dextran-coated MNPs was synthesized and isolated. Thrombolysis by the conjugate was studied under a magnetic field in a rat arteriovenous shunt thrombosis model. The magnetic field was generated by two AlNiCo permanent magnets around the site of thrombus. The magnetic field enhanced the thrombolytic efficacy of the conjugate by 5-fold over urokinase with no reduction in plasma fibrinogen and little prolonged bleeding time. It suggested that thrombolysis had been specifically directed to the desired site by the magnetic carrier under the magnetic field. Additionally, the conjugate had a longer half-life than urokinase in circulation.

Aims: To investigate protein and gene expressions of chemokine subtypes CCR3, CCR2A and their respective ligands macrophage inflammatory protein 1-alpha (MIP-1alpha), monocyte chemotactic protein-1 (MCP-1) in the normal mouse central nervous system (CNS) and in the hippocampus at different time points during and after pilocarpine-induced status epilepticus (PISE)., Methods: CCR3 and MIP-1alpha protein expressions were mapped in the mouse CNS. The protein and gene expressions of CCR3 and CCR2A and their respective ligands MIP-1alpha, MCP-1 in the hippocampus were studies by immunocytochemical and quantitative real-time RT-PCR during and after PISE., Results: CCR3 and MIP-1alpha gene expression and immunopositive neurones were broadly distributed in the CNS. CCR3 and CCA2A gene and their protein expression were downregulated in the hippocampus at 1 h during PISE. The protein expression of MIP-1alpha, MCP-1 decreased but gene expression increased at 2 h during PISE. In the hilus of the dentate gyrus, significant reduction of the numbers of CCR3, CCR2A, MCP-1 immunopositive neurones occurred from 1 h during to 2 months after PISE, but the number of MIP-1alpha neurones reduced from 2 h during to 2 months after PISE. Induced expression of CCR3 at 1 week, CCR2A, MCP-1 or MIP-1alpha at 1 week and 2 months after PISE was found in reactive astrocytes. MCP-1 was also demonstrated in the blood vessels of the hippocampus at 2 months after PISE., Conclusions: CCR3 and MIP-1alpha may play important functional roles in the mouse brain. The downregulation of CCR3, CCR2A, MIP-1alpha and MCP-1 in the hippocampal neurones at the acute stage during and after PISE may weaken the neuroprotective mechanisms. However, induced expression of MCP-1 in hippocampal blood vessel may be related to changes in permeability of the blood-brain barrier during epileptogenesis.

The recently arising antithrombin drug, angiomax, was successfully conjugated with a 5'-amino oligonucleotide through click chemistry. This oligo-angiomax conjugate was assembled into a two-dimensional DNA lattice with other oligonucleotides together. Besides the plane sheet of DNA lattices, an interesting angiomax-involved DNA tubing structure, constructed by 40 to 50 angiomax stripes which are parallel to the longitudinal axis of the tube, was also imaged. After incubation of thrombins with the angiomax-involved DNA lattice, the binding of thrombins to arrayed angiomax peptides was observed. Finally a chromogenic substrate bioassay was employed to estimate the antithrombin activities as assembled oligo-angiomax DNA lattice approximately 1.1, oligo-angiomax approximately 2.7 angiomax. The functionalized DNA lattices have the potential to be used as a powerful platform for investigation of biomolecular interactions such as drug-protein, protein-protein, DNA-RNA, and DNA-protein interactions in the nano- and subnanoscales.

Objective: To investigate whether proteasome inhibitor MG-132 induces apoptosis of human erythroleukemia cell line K562 and possible mechanisms., Methods: K562 cells were incubated with RPMI 1640 and exposed to 0, 1, 5, 10, 15 micromol/L of MG-132 for 24 hrs, respectively. The apoptosis of cells were detected by fluorescence microscope, DNA fragments and flow cytometry. The NF-kappaB mRNA expression was quantified by reverse transcription-polymerase chain reaction (RT-PCR). Expression of NF-kappaB and caspase-3 was semiquantitatively analyzed with SABC techniques. Caspase-3 activities were measured with a colorimetric method., Results: The growth of K562 cells was inhibited and the apoptosis of the cells increased after MG-132 treatment in a dose-dependent manner. After 24 hrs of 15 micromol/L MG-132 treatment, the percentage of apoptotic cells (26.5+/-0.6%) increased significantly when compared with the untreated controls (1.2+/-0.1%) (P<0.01). MG-132 treatment decreased the mRNA and protein expression of NF-kappaB, and increased the protein expression of caspase-3., Conclusions: MG-132 can induce apoptosis of human erythroleukemia cell line K562 through the down-regulation of NF-kappaB expression and up-regulation of caspase-3 expression.

A proof-of-concept study demonstrated the feasibility of a novel gel-pad microarray on porous silicon chips, by initiation of an atom transfer radical propagation (ATRP) polymerisation of (polyethylene glycol) methacrylate (PEGMA) with surface Si-H species, stepwise chemical conversions of the gel membrane to an NTA-Ni2+/histidine-tagged protein system, and matrix-assisted laser desorption/ionisation mass spectroscopy (MALDI MS) and fluorescence detections.

Aim: To investigate the effectiveness of pre-emptive preperitoneal infiltration of 0.5% Bupivacaine in postoperative pain control in laparoscopic total extraperitoneal (TEP) hernioplasty., Method: We conducted a prospective randomized controlled three-arm study. Consecutive patients aged 18-75 years with reducible groin hernia undergoing laparoscopic TEP hernioplasty in our day surgery unit were recruited. They were randomized into three groups. The pre-emptive Bupivacaine group (PBU) received infiltration of 10 ml 0.5% Bupivacaine to port sites before skin incision and another 10 ml to the preperitoneal space immediately after the creation of a first working port before further dissection. The standard Bupivacaine group (SBU) received infiltration of Bupivacaine after mesh placement, while in the control group (CO), the same amount of saline was injected instead. All operations were performed with the same surgical technique by a single surgeon. Postoperative pain was recorded at 2, 6, and 24 h by an independent assessor using a 0-10 verbal rating scale. The operative time, complications, and analgesia consumption, as well as patient satisfaction were also measured., Results: Ninety patients were randomized into three groups of 30. There were no differences in age, American Society of Anesthesiologists (ASA) risk classification, number of bilateral diseases, and operation time. The pain score was significantly reduced by Bupivacaine compared with control patients, while pre-emptive Bupivacaine was better than standard Bupivacaine. Fourteen out of ninety patients (15.6%) had seroma formation. All patients were discharged on the same day of operation., Conclusion: Pre-emptive preperitoneal infiltration of 0.5% Bupivacaine significantly reduces postoperative pain in laparoscopic TEP hernioplasty.

Hospitals and health care providers tend to get involved in exaggerated and fraudulent medical claims initiated by national insurance schemes. The present study applies data mining techniques to detect fraudulent or abusive reporting by healthcare providers using their invoices for diabetic outpatient services. This research is pursued in the context of Taiwan's National Health Insurance system. We compare the identification accuracy of three algorithms: logistic regression, neural network, and classification trees. While all three are quite accurate, the classification tree model performs the best with an overall correct identification rate of 99%. It is followed by the neural network (96%) and the logistic regression model (92%).

While it is generally accepted that phospholipase C (PLC) and protein kinase C (PKC) are down-stream proteins involved in metabotropic glutamate receptor 5 (mGluR5)-related signal transduction, we still do not know which subtype of PLC or PKC is specifically regulated after mGluR5 activation. In the present study in mGluR5 wild-type (mGluR5+/+) mice, we showed induced PKCbeta2 or PKCgamma expression at the border between the stratum oriens and alveus (O/A border) at 2h during pilocarpine induced status epilepticus (SE), and in the stratum pyramidale in CA1 area at 1 day after pilocarpine induced SE; at 1 day, induced expression of PLCbeta4 in the stratum pyramidale of CA1 area was observed. Furthermore, double labeling revealed the co-localization of induced PKCbeta2 or PKCgamma with mGluR5 or with induced PLCbeta4 in the stratum pyramidale of CA1 area. These induced expression, however, were not found in mGluR5 mutant (mGluR5-/-) mice. It suggests that induced PLCbeta4-PKCbeta2/PKCgamma at 1 day after pilocarpine induced SE in pyramidal neurons or PKCbeta2 or PKCgamma in interneurons at O/A border at 2h during pilocarpine induced SE may be specifically linked to the activation of mGluR5. When compared to mGluR5+/+ mice, significant shorter latency (from pilocarpine injection to the occurrence of status epilepticus) and maintenance period (from beginning to the end of status epilepticus) for status epilepticus in mGluR5-/- mice were also demonstrated. It is possible that mGluR5 may play a negative role in initiation of status epilepticus by interacting with muscarinic acetylcholine receptor in mGluR5+/+ mice.

Objective: To investigate the expression of cyclin A protein in childhood acute leukemia (AL) and its significance., Methods: By using Western blotting analysis, cyclin A protein in bone marrow mononuclear cells from 47 newly diagnosed AL children and 33 non-hematological malignancy children was detected., Results: The expression of cyclin A in AL group (0.38 +/- 0.20) was higher than that in control group (0.03 +/- 0.15) (P < 0.01). The expression of cyclin A in high risk acute lymphocyte leukemia (ALL) group (HR-ALL) (0.62 +/- 0.38) was higher than that in standard risk ALL group (SR-ALL) (0.33 +/- 0.33) (P < 0.05). The expression of cyclin A in WBC > or = 50 x 10(9)/L group and in WBC < 50 x 10(9)/L group was (0.64 +/- 0.36) and (0.39 +/- 0.38), respectively (P < 0.05). Eight (44.4%) out of 18 patients with positive cyclin A expression achieved complete remission (CR). The CR rate was lower than that of patients with negative cyclin A expression (100%) (P < 0.01)., Conclusions: The higher expression of cyclin A may predict a poor prognosis for childhood ALL.

This study was undertaken to explore the anatomic features and adjacent relationships of the pineal region in thin coronal sections. After CT and MR examination verifying no brain lesions, one normal cadaver head was selected for this study from three Chinese adult male cadavers. After being embedded and frozen, the head was sliced into serial sections at 0.1 mm intervals in the coronal plane with SKC 500 computerized freezing milling machine. Then the serial coronal sections were photographed by a high-resolution digital camera and saved in the computer. Subsequently, the anatomic structures of the pineal region on the thin coronal sections were investigated and correlated with in vivo MR images, which were obtained from ten normal Chinese male adult volunteers by a 3.0 T GE scanner. The base lines of the sectioning and the MR scan were all perpendicular to the AC-PC line. A total of 355 coronal sections and 21-23 in vivo coronal MR images related with the pineal region were obtained, respectively. From anterior to posterior, the shape of the pineal region changed from an inverted triangle to a trapezoid and a triangle gradually, and the anatomic details could be depicted clearly in the thin sectional anatomy images in sub-millimeter. Via the comparison, some micro-anatomic structures of the pineal region that cannot be discriminated clearly or missed in the thick sections or MR images were identified. The contrast of the computerized freezing milling technique with the MRI enhanced our ability to comprehend the complex anatomy of the pineal region and to improve the imaging diagnosis and surgical treatments of minute diseases in this region.

Introduction: The aim of this retrospective study was to assess the feasibility and outcome of day case thyroidectomy in an ambulatory surgery centre in Hong Kong., Methods: Patients with day case thyroidectomy carried out between July 2005 and December 2006 were retrospectively reviewed. Day surgery was offered to patients satisfying the selection criteria for day case and having from benign unilobular thyroid disease., Results: Fifty patients had hemithyroidectomy carried out during the study period. There were 6 men and 44 women and the mean (standard deviation SD) age was 45.6 years (7.4 years). All patients were American Society for Anesthesiologists grade I (76%) or II (34%). The mean (SD) operative time was 79.5 min (17 min). Twelve patients had episodes of postoperative nausea and vomiting. The mean (SD) analgesic requirement was 0.7 tablets (0.5 tablets) of combination acetaminophen and phenyltoloxamine citrate before discharge. The mean (SD) time to discharge was 7.5 h (0.7 h). The overall discharge rate was 98% and the complication rate was 8%. One patient was observed overnight because of postoperative haematoma. One patient had recurrent laryngeal nerve injury. There were no unplanned readmissions postoperatively. Three patients had unsuspected thyroid malignancy on histopathology., Conclusion: This study showed the feasibility and safety of day case thyroidectomy. The setting was not associated with any increase in morbidity or mortality and has the potential in reducing hospital costs.

The GroEL/GroES chaperonin system of Escherichia coli forms a nano-cage allowing single protein molecules to fold in isolation. However, as the chaperonin can also mediate folding independently of substrate encapsulation, it remained unclear whether the folding cage is essential in vivo. To address this question, we replaced wild-type GroEL with mutants of GroEL having either a reduced cage volume or altered charge properties of the cage wall. A stepwise reduction in cage size resulted in a gradual loss of cell viability, although the mutants bound non-native protein efficiently. Strikingly, a mild reduction in cage size increased the yield and the apparent rate of green fluorescent protein folding, consistent with the view that an effect of steric confinement can accelerate folding. As shown in vitro, the observed acceleration of folding was dependent on protein encapsulation by GroES but independent of GroES cycling regulated by the GroEL ATPase. Altering the net-negative charge of the GroEL cage wall also strongly affected chaperonin function. Based on these findings, the GroEL/GroES compartment is essential for protein folding in vivo.

With the mouse pilocarpine model of temporal lobe epilepsy (TLE), we showed a progressive loss of both principal cells and calbindin (CB)-, calretinin (CR)-, and parvalbumin (PV)-immunopositive interneurons in layers II-III of lateral entorhinal cortex (LEnt) from 2 months to 1 year after pilocarpine-induced status epilepticus (PISE). In the efferent pathway of LEnt, more Phaseolus vulgaris leucoagglutinin (PHA-L)-labelled en passant and terminal boutons with larger diameters were shown in the hippocampus and subiculum; in the prefrontal, piriform, and perirhinal cortices; and in the amygdaloid complex in experimental mice at the two time points compared with the control after iontophoretical injection of an anterograde tracer PHA-L into the LEnt. Furthermore, the numbers of CB- or CR-immunopositive neurons contacted by PHA-L-labelled en passant and terminal boutons decreased in most of these areas at 2 months or 1 year after PISE. In the afferent pathway of LEnt, the numbers of retrogradely labelled neurons were reduced significantly in the ipsilateral piriform cortex and endopiriform nucleus at 2 months and 1 year and in the reuniens thalamic nucleus only at 1 year after injection of a retrograde tracer cholera toxin B subunit (CTB) into the LEnt. The percentages of the number of CTB and CB or CR double-labelled neurons of all the retrogradely labelled neurons were also decreased in the reunions thalamic nucleus at 1 year after PISE. It is concluded that both cytoarchitectonic change and reorganization of afferent and efferent pathways in LEnt may be involved in the occurrence of TLE., (2007 Wiley-Liss, Inc.)

To provide practical anatomic data for the imaging diagnosis and surgical treatment of adrenal disease, we investigated the anatomy of the adrenal gland and its relationships to regional structures using 31 sets of serial coronal sections of upper abdomen of Chinese adult cadavers and correlated coronal magnetic resonance (MR) images of ten upper abdomens of adult healthy volunteers and coronal reconstructed multislice spiral computed tomography (MSCT) images of five patients without lesions in the adrenal gland. The adrenal glands were visualized mainly on the successive coronal sections between 18 mm anterior to the posterior margin of inferior vena cava and 24 mm posterior to the posterior margin of inferior vena cava. In general, the left adrenal gland was visualized two sections earlier than the right adrenal gland. On the plane through the anterior parts of bilateral renal hili (A18), the appearance rate of bilateral adrenal glands was 100%, and the maximal measurements of bilateral adrenal glands were visualized. The length, width, thickness of right adrenal body, thickness of medial limb and lateral limb were, respectively, 34.02 +/- 2.12 mm, 10.91 +/- 0.89 mm, 5.82 +/- 0.26 mm, 2.78 +/- 0.08 mm, 2.62 +/- 0.06 mm, whereas the measurements of left adrenal gland were 28.31 +/- 2.46 mm, 18.40 +/- 1.06 mm, 6.84 +/- 0.24 mm, 3.02 +/- 0.08 mm, 2.86 +/- 0.07 mm, respectively. The coronal plane has superior advantage in showing the bilateral adrenal glands. The shapes of adrenal glands are various, whereas the range of adrenal thickness is quite narrow. The thickness of adrenal medial and lateral limbs, especially the thickness of lateral limb are useful for the diagnosis of the bilateral adrenocortical disease.

Microarray technology has made it possible to simultaneously study the abundance, interactions, and functions of potentially tens of thousands of biological molecules. From its earliest use in DNA microarrays, where only nucleic acids were captured and detected on the arrays, applications of microarrays now extend to those involving biomolecules such as antibodies, proteins, peptides, and carbohydrates. In contrast to the relative robustness of DNA microarrays, the use of such chemically diverse biomolecules on microarray formats presents many challenges in their fabrication as well as application. Among the many methods that have been proposed to overcome these challenges, DNA-directed assembly (DDA) has emerged as a promising strategy for the high sensitivity and multiplexed capture and detection of various analytes. In this review, we explore the challenges faced during the design, fabrication, and utilization of protein microarrays and highlight how DDA strategies, together with other recent advances in the field, are accelerating the development of platforms available for protein microarray applications.

Disturbance of circadian gene regulation might contribute to behavioral and psychological symptoms in dementia patients. This study was to evaluate the CpG island methylation status on the circadian gene promoters in dementia patients. We conducted a set of methylation specific polymerase chain reaction (mPCR) followed by nucleotide sequencing to analyze the methylation status within the promoters of nine circadian-related genes, including PER1, PER2, PER3, CRY1, CRY2, CLOCK, BMAL1, TIM and CK1epsilon, in the genomic DNA from the peripheral blood leukocytes of 80 dementia patients and 80 age- and gender-matched controls. A total of seven dementia patients (7/80) had CpG island methylation in the circadian genes and none of the controls had methylation. There were three and four patients had CpG island methylation on the promoters of PER1 and CRY1, respectively. Dementia with Lewy body (DLB) patients had the significantly highest frequency of circadian gene CpG island methylation (35.7%). It suggested that epigenetic methylation of circadian gene was more prevalent in dementia patients, especially for the DLB patients. The significance of circadian gene methylation in clinical behavior/sleep disturbance in dementia patients needs further study.

The GroEL/GroES chaperonin system mediates protein folding in the bacterial cytosol. Newly synthesized proteins reach GroEL via transfer from upstream chaperones such as DnaK/DnaJ (Hsp70). Here we employed single molecule and ensemble FRET to monitor the conformational transitions of a model substrate as it proceeds along this chaperone pathway. We find that DnaK/DnaJ stabilizes the protein in collapsed states that fold exceedingly slowly. Transfer to GroEL results in unfolding, with a fraction of molecules reaching locally highly expanded conformations. ATP-induced domain movements in GroEL cause transient further unfolding and rapid mobilization of protein segments with moderate hydrophobicity, allowing partial compaction on the GroEL surface. The more hydrophobic regions are released upon subsequent protein encapsulation in the central GroEL cavity by GroES, completing compaction and allowing rapid folding. Segmental chain release and compaction may be important in avoiding misfolding by proteins that fail to fold efficiently through spontaneous hydrophobic collapse.

Freshwater crabs (Sinopotamon denticulatum) were examined for metacercariae. Cats and dogs were also examined for Paragonimus infection. Questionnairing was carried out on health knowledge and behaviors among local residents in a village of Baokang County, Hubei Province. Results showed that the infection rate of Paragonimus skrjabini metacercariae in Sinopotamon denticulatum was 20.5% (46/214), with 15.6% (20/128) in a mining area and 30.2% (26/86) for the non-mining area respectively (chi2 = 6.5, P < 0.05). The prevalence in cats and dogs was 25.0% (6/24) and 17.6% (6/34) respectively (chi2 = 0.46, P > 0.05). Questionnairing showed that dogs and cats were with the habit of foraging and defecating at streams and children had the habits of eating raw or under-cooked crabs. The natural and ecological environments are in favor of the life cycle of P. skrjabini.

Toluene, an industrial organic solvent, is voluntarily inhaled as drug of abuse. Toluene has been shown to inhibit the nicotinic acetylcholine receptors. Nicotinic receptors play an important role in brain development during brain growth spurt and early adolescence. The long-term effects of neonatal and adolescent toluene exposure on behavioral responses to nicotine in early adulthood were compared. Sprague-Dawley male and female rats were treated with toluene (500 mg/kg, ip) or corn oil daily over postnatal day (PN) 4-9 or 25-30. Nicotine-induced hypothermia, antinociception, and seizure activity were examined during PN 56-60. Toluene exposure during the brain growth spurt, but not adolescence, reduced the behavioral responses to nicotine in young adult rats. However, the levels of alpha4, alpha7, and beta2 nicotinic receptors were not altered in the frontal cortex, striatum, thalamus, hippocampus, and cerebellum by neonatal toluene exposure. These results indicate that toluene exposure during the brain growth spurt produces long-term changes in nicotine sensitivity, which may be unrelated to the total expression levels of alpha4, alpha7, and beta2 nicotinic receptors. The alterations in nicotine sensitivity may be related to the neurobehavioral disturbance associated with fetal solvent syndrome.

Protein microarray offers a means for high-throughput profiling of cellular proteins to provide insights into the mechanisms of biological processes. This study describes the design and fabrication of a robust platform, spatially addressable protein array (SAPA), by exploring the specificity of ssDNA hybridization for self-assembly of semi-synthetic ssDNA-antibody conjugates which capture antigens from complex biological samples. This approach does not involve the direct immobilization of antibodies nor antigen, but instead captures the target antigens in the solution phase followed by self-directed assembly of the complex onto the surface. In an effort to optimize the platform, the effects of surface chemistry, nonspecific protein adsorption, facile preparation, and purification of ssDNA-conjugated antibody and capture of the antigen from a complex biological sample such as cell lysate were examined. This platform allowed antigen detection in cell lysate with high sensitivity (1 pM). The method described herein can be extended to the high-throughput detection of other interacting molecules in solution phase and their subsequent assembly onto any substrate.

By intravenous administration of group I metabotropic glutamate receptor antagonists at 1 or 2h during pilocarpine induced status epilepticus (PISE), we showed that mGluR1 antagonists AIDA or LY367385 (at dosages ranging from 25 to 200mg/kg), mGluR5 antagonists SIB1757 (at dosages ranging from 25 to 200mg/kg), SIB1893 (from 25 to 100mg/kg), MPEP (from 25 to 100mg/kg) injected at 1 or 2h during PISE were ineffective in controlling status epilepticus (SE). However, when administered at 1h during PISE, MPEP at 200mg/kg, combination of MPEP (200mg/kg) with MK801 (0.1mg/kg) or with MK801 (0.1mg/kg) and diazepam (0.5mg/kg), combination of SIB1893 (200mg/kg) with MK801 (0.1mg/kg) could effectively control behavioral SE, and were neuroprotective. In particular, the combination of MPEP with MK801 and diazepam could stop both behavioral SE and electrical SE (under EEG monitoring) within a few minutes after the administration. HPLC study showed that a high level of MPEP was maintained in the blood and its metabolism rate was slow in experimental mice with PISE. We therefore concluded that the combination of MPEP (200mg/kg) with MK801 (0.1mg/kg) and diazepam (0.5mg/kg) could effectively stop SE and its subsequent neuronal loss in the hippocampus when administered 1h during PISE. It may provide a new approach to effectively control intractable SE.

Protein microarrays are promising tools that can potentially enable high throughput proteomic screening in areas such as disease diagnosis and drug discovery. A critical aspect in the development of protein microarrays is the optimization of the array's surface chemistry to achieve the high sensitivity required for detection of proteins in cell lysate and other complex biological mixtures. In the present study, a high-density antibody array with minimal nonspecific cellular protein adsorption was prepared using a glass surface coated with a poly(propyleneimine) dendrimer terminated with carboxyl group (PAMAM-COOH). The carboxyl-terminated dendrimer-modified surface has almost similar nonspecific cellular protein adsorption when compared to an inert PEG-modified surface. In addition, the multiple functional sites available for reaction on the dendrimer surface facilitated high-density immobilization of antibodies and efficient capture of bioanalytes. Various molecules were tested for their ability to block or deactivate the reactive carboxyl surface after antibody immobilization to further reduce the nonspecific binding. A short oligoethylene glycol (NH2-d4-PEG-COOH), was found to significantly improve the signal-to-noise ratio of the assay, resulting in higher sensitivity. The properties and functional qualities of the various surfaces were characterized by contact angle and AFM measurements. Nonspecific protein adsorption and protein immobilization as a function of dendrimer generations and sensitivity of antigen capturing from a buffer (1 pM) as well as from the complex cell lysate (10 pM) system were examined. Our detailed experimental studies demonstrated a facile method of preparing surfaces with high protein loading and low nonspecific protein binding for the development of high sensitivity protein microarrays.

Purpose: We sought to investigate the anticonvulsive and neuroprotective effect of a selective metabotropic glutamate receptor 8 (mGluR8) agonist (S)-3,4-dicarboxyphenylglycines (S-3,4-DCPG) on pilocarpine-induced status epilepticus (PISE) and subsequent loss of hilar neurons in the dentate gyrus after systemic (intravenous) or local (intracerebroventricular) administration. We compared the difference in granular cell responses after paired-pulse stimulation of the perforant pathway and the sensitivity to local injection of S-3,4-DCPG into the stratum granulosum in the control and mice at 2 months after PISE., Methods: We used intravenous, intracerebroventricular, or intrahippocampal administration of S-3,4-DCPG to mice with status epilepticus or temporal lobe epilepsy and neurophysiologic recording of somatic field excitatory postsynaptic potential (sfEPSP) and population spike (PS) of granular cells in response to perforant-pathway stimulation or S-3,4-DCPG treatment., Results: Intracerebroventricular (1.91 micromol) but not systemic administration of S-3,4-DCPG (at doses of 12.5, 50, 100, 200, 400, 800, and 1,200 mg/kg) could control PISE with no neuroprotective effect. In epileptic mice, mGluR8-mediated inhibition of fEPSPs was reduced significantly in granular cell bodies., Conclusions: At doses ranging from 12.5 to 1,200 mg/kg, intravenous administration of S-3,4-DCPG may not be effective in controlling status epilepticus. Down-regulation of mGluR8 may be related to reduced S-3,4-DCPG-mediated inhibition and the subsequent occurrence of spontaneously recurrent seizures.