Integrated contra-directionally coupled chirped Bragg grating waveguide with a linear group delay spectrum.

Due to the advantages of low propagation loss, wide operation bandwidth, continuous delay tuning, fast tuning speed, and compact footprints, chirped Bragg grating waveguide has great application potential in wideband phased array beamforming systems. However, the disadvantage of large group delay error hinders their practical applications. The nonlinear group delay spectrum is one of the main factors causing large group delay errors. To solve this problem, waveguides with nonlinear gradient widths are adopted in this study to compensate for the nonlinear effect of the grating apodization on the mode effective index. As a result, a linear group delay spectrum is obtained in the experiment, and the group delay error is halved.

Design of compact dual-mode photoelectric modulator with high process tolerance based on vanadium dioxide.

Opto-electro modulators with nanometer-scale footprint are indispensable in integrated photonic electronic circuits. Due to weak light-matter interactions and limits of micro-nano fabrication technology, it is challenging to shrink a modulator to subwavelength size. In recent years, hybrid modulators based on surface plasmons have been proposed to solve this problem. Although the introduced high lossy surface plasmons provide large modulation depth, the polarization selectivity limits its application. Toward this end, in this paper, we present a design of an ultra-compact vanadium oxide (VO2)-based plasmonic waveguide modulator for both transverse electric (TE) and transverse magnetic (TM) modes. The device consists of two silicon tapers and a silicon waveguide embedded with a VO2 wedge. When electrical signals put on the device change the phase of VO2 from a metal to an insulator, the output optical signals along the waveguide are significantly modulated. For a 1.5 µm length modulator operating at 1.55 µm wavelength, the extinction ratio is 11.62 dB for the TE mode and 8.86 dB for the TM mode, while the insertion loss is 4.31 dB for the TE mode and 4.12 dB for the TM mode. Furthermore, the proposed design has excellent tolerance for fabrication process error, which greatly increases the yield rate of products and indicates a promotable application prospect.

[Effect of ouabain on intracellular Ca(2+) concentration in rat vascular smooth muscle cells in vitro].

OBJECTIVE: To explore the effect of ouabain on intracellular Ca(2+) concentration ([Ca(2+)]i) in thoracic aorta vascular smooth muscle cells (VSMCs) in vitro. METHODS: Primary SD rat thoracic aorta VSMCs were cultured by tissue adherent method and identified by immunochemistry. The binding ability between ouabain and VSMCs was detected by autoradiography, and fluo 3-AM (a Ca(2+) fluorescent probe) was employed to investigate whether ouabain affected VSMCs within a short period of time. The effect of a truncated fragment of the sodium pump α2 subunit was assayed in antagonizing the effect of ouabain on [Ca(2+)]i in the VSMCs. RESULTS: Within the concentration range of 0.1-100 nmol/L, ouabain was found to dose-dependently bind to the VSMCs. Different concentrations of ouabain (0-3200 nmol/L) caused a transient, dose-dependent increase in [Ca(2+)]i in the VSMCs, which was antagonized by the application of the truncated fragment of sodium pump α2 subunit. CONCLUSIONS: Elevations in [Ca(2+)]i in the VSMCs can be the cytological basis of high ouabain-induced hypertension. The truncated fragment of the sodium pump α2 subunit can antagonize ouabain-induced increase of [Ca(2+)]i in the VSMCs, which provides a clue for understanding the pathogenesis of and devising a therapeutic strategy for high ouabain-induced hypertension.

[Preparation and characterization of polyclonal antibodies against rat sodium pump alpha 2 subunit M1-M2 extra membrane fragment].

OBJECTIVE: To prepare polyclonal antibodies against sodium pump alpha 2 subunit M1-M2 extramembrane fragment (NKAα2 EM1) for studying the pathogenesis of hypertension. METHODS: According to the GenBank data, the amino acid sequence of NKAα2 EM1 was obtained and the target peptide (LAAMEDEPSNDN) was synthesized using a peptide synthesizer with Fmoc method and purified with high-performance liquid chromatography. The synthesized peptide was then coupled to KLH for immunizing New Zealand white rabbits for 4 times to obtain the antiserum. The IgG antibodies against the synthetic peptide, after affinity purification with Protein A, were used for detecting NKAα2 EM1 expression in rat aortic vascular smooth muscle cells by enzyme-linked immunosorbent assay and immunocytochemistry (ICC). RESULTS: The synthesized peptide fragment , which consisted of 13 amino acid residues including one derivatized cysteine residue in the N-terminal (LAAMEDEPSNDN-C), had a theoretical relative molecular mass of 1408.48 D with a measured relative molecular mass of 1407.90 D and a purity exceeding 85.5%. The titer of the antiserum was more than 1:512 000, and the purified IgG antibody concentration was 0.965 mg/ml after purification with Protein A. At a 1:1000 dilution (final concentration of 1 µg/ml), the titer of the purified IgG antibody was more than 1:256 000. The purified IgG antibody could be used at 1:100 to 1:200 dilutions for for immunocytological examination of formalin-fixed cells. CONCLUSION: The anti-NKAα2 EM1 polyclonal antibodies obtained can be used in ELISA and immunocytochemistry for detecting the sodium pump alpha 2 subunit in formalin-fixed tissue or cells to facilitate investigation of the relationship between sodium pump and hypertension.

TLR2-ICAM1-Gadd45α axis mediates the epigenetic effect of selenium on DNA methylation and gene expression in Keshan disease.

Keshan disease (KD) is a fatal dilated cardiomyopathy with unknown etiology, and selenium deficiency is considered the main cause of KD. Several observations implicate a role for altered DNA methylation in selenium deficiency-related diseases. The aim of the present study was to investigate the epigenetic effects of selenium (Se) on DNA methylation and gene expression in Keshan disease. Using methylated DNA immunoprecipitation chip (MeDIP-Chip) and quantitative RT-PCR, we identified two inflammatory-related genes (TLR2 and ICAM1) that were differentially methylated and expressed between normal individuals and KD patients. Results from DNA methylation profile between KD patients and normal individuals showed that selenium deficiency decreased methylation of CpG islands in promoter regions of TLR2 and ICAM1 and upregulated messenger RNA (mRNA) and protein levels of TLR2 and ICAM1. In rat animal model of Keshan disease, selenite treatment could increase TLR2 and ICAM1 promoter methylation, suppress these genes expression, and reduce infiltration of myocardial inflammatory cells. In cell culture model of Keshan disease, we found 5-Aza-dC (DNMT1 inhibitor) treatment in the presence of selenium-reduced mRNA and protein levels of DNMT1 regardless of TLR2 and ICAM1 promoter methylation status and expression levels of these genes. Selenite treatment suppressed the expression of the Gadd45α, TLR2, and ICAM1 in a concentration-dependent manner, while selenium deficiency increased the expression of the Gadd45α, TLR2, and ICAM1 and decreased TLR2 and ICAM1 promoter methylation level in a time-dependent manner. Our results revealed that TLR2-ICAM1-Gadd45α axis might play an important role in gene-specific active DNA demethylation during inflammatory response in myocardium.

Oxidized low-density lipoprotein induces inflammatory responses in cultured human mast cells via Toll-like receptor 4.

BACKGROUND/AIMS: Oxidized low-density lipoprotein (ox-LDL) is a powerful atherogen. Toll-like receptor 4 (TLR4) has a pathophysiological role in regulating inflammatory responses and atherosclerosis. Mast cells can infiltrate into the atheromatous plaque and secrete various pro-inflammatory cytokines, which significantly amplify the atherogenic processes and promote plaque vulnerability. Small interfering RNA (siRNA) is an effective method to silence the target genes. We evaluated whether ox-LDL-induced inflammation depended in part on the activation of TLR4-dependent signaling pathways in a cultured human mast cell line (HMC-1). METHOD: HMC-1 cells were cultured, and treated with ox-LDL, TLR4-specific siRNA, or inhibitors of phosphorylation of mitogen-activated protein kinase (MAPKs), and nuclear factor-κB (NF-κB), a critical mediator of inflammation. The expression of monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6) was measured subsequently. RESULTS: Ox-LDL increased the expression of TLR4 and secretion of MCP-1, TNF-α and IL-6. Moreover, ox-LDL stimulated the translocation of NF-κB, from the cytoplasm to nucleus. Additionally, phosphorylation of MAPK was greatly increased. These ox-LDL-induced alterations were significantly attenuated by pretreatment with TLR4-specific siRNA. CONCLUSION: Ox-LDL induced inflammatory responses in cultured HMC-1 cells including NF-κB nuclear translocation and phosphorylation of MAPKs, a process mediated in part by TLR4.

The new role of LOX-1 in hypertension induced neuronal apoptosis.

Lectin-like oxidized low-density lipoprotein (oxLDL) receptor-1 (LOX-1) was originally identified as a receptor for oxLDL predominantly expressed in endothelial cells. Recently up-regulation of LOX-1 has been implicated in oxidative stress and cell apoptosis in many cell types. However, LOX-1 expression in neurons or regulation of neuronal apoptosis by LOX-1 has not been reported. To investigate the possible roles of LOX-1 in hypertension induced brain damage, we examined the distribution of LOX-1 in cortex and hippocampus and compared its expression in 32-week-old SHR and WKY rats. Immunofluorescence revealed that LOX-1 positive cells were located principally at the cortex involved in sensory information processing and were mainly expressed in neurons. We also found up-regulated mRNA expression of LOX-1, Bax and caspase-3 and down-regulated mRNA expression of Bcl-2 in SHR group. Compared with WKY group, SHR group showed increased LOX-1 positive cells and TUNEL positive cells. Furthermore, double-labeling method indicated that LOX-1 expression was colocalized with TUNEL positive cells, which means that LOX-1 expression was involved in hypertension related cell apoptosis. These findings indicated that LOX-1 expression was up-regulated in the cortex of SHR and its expression has implication in neuronal apoptosis. Elevated Bax/Bcl-2 ratio may be involved under this event.

Applying the extensor digitorum reflex to neurological examination.

AIM: To determine the value of the extensor digitorum reflex in neurologic examination. METHODS: The extensor digitorum, biceps, and brachioradialis reflexes were elicited in 65 patients with hemiplegia and upper-limb paralysis and in a control group of 120 apparently healthy people. Reflexes were elicited by both conventional means and a new method for the extensor digitorum reflex. The sensitivity and specificity of the extensor digitorum reflex were compared with that of the conventional biceps and brachioradialis reflexes to evaluate the value of the extensor digitorum reflex for neurologic examination. RESULTS: The sensitivity of the extensor digitorum, biceps, and brachioradialis reflexes were 93.65%, 90.48%, and 90.48%, respectively. The specificity of the extensor digitorum, biceps, and brachioradialis reflexes were 95.83%, 94.17%, and 93.33%, respectively. The diagnostic efficacies of the extensor digitorum, biceps, and brachioradialis reflexes were 95.08%, 92.90%, and 91.26%, respectively. There were no significant differences (p > 0.05) in sensitivity, specificity, or accuracy among the extensor digitorum, biceps or brachioradialis reflexes in neurologic examination. CONCLUSIONS: The extensor digitorum reflex is a sensitive and useful deep tendon reflex and is suitable for widespread use in neurological examination.

[Expression and purification of truncated fragment of extracellular segment of sodium pump alpha3 subunit in Escherichia coli by in-fusion technology].

OBJECTIVE: To construct prokaryotic expression system for expressing, purifying and identifying truncated fragment of extra-cellular segment of sodium pump alpha3 subunit with pGEX-6P-1 GST gene fusion system in Escherichia coli by in-fusion technology. METHODS: According to the conservative sequence of M1-M2 and M3-M4 extra-cellular gene fragments of sodium pump a3 subunit, which published in GenBank, a serial of primers and gene fragments was designed, and directly synthesized to fuse the above two gene fragments. The fusion gene was fused with gene-specific primers by PCR, and then fusion gene fragment was fused into the single stranded homology regions of vector pGEX-6P-1 by in-fusion cloning to construct recombinant vector pGEX-Trf-alpha3 (Truncated fragment of extracellular segment of sodium pump alpha3 subunit, Trf-alpha3). After DH10bac was transferred with it, the pGEX-Trf-alpha3 plasmid was purified and identified by PCR and sequenced. Then the recombinant plasmid pGEX-Trf-alpha3 was expressed in E. coli BL21 cells, inducted by IPTG. GST-Trf-alpha3 fusion protein was purified with Glutathione Sepharose 4B purifying system and analyzed by SDS-PAGE. RESULTS: The results of PCR and sequencing demonstrated that the M1-M2 and M3-M4 extra-cellular gene was inserted in plasmid pGEX-6P-1 vector successfully. And the sequence was correct. Protein sequence analysis showed that the GST-Trf-alpha3 fusion protein was consisted of 262 amino-acid residues. Relative molecular mass in theory was 33.22 X 10(3). The amount of recombinant protein was 10% of the total bacteria protein. The soluble fusion protein was about 80.8%. After affinity purification, the purity of GST-Trf-alpha3 fusion protein was over 95%. There was some extent binding activity between GST-Trf-alpha3 fusion protein and ouabain, but the activity was very low. CONCLUSION: Prokaryotic expression system for expressing truncated fragment of extra-cellular segment of sodium pump alpha3 subunit with pGEX-6P-1 GST gene fusion system in Escherichia coli by in-fusion technology had been constructed. The purified method had also established. High purified GST-Trf-alpha3 fusion protein was obtained. These have found the foundation of further study on its biological function and potential pharmacology function.

[Binding activity of polypeptide containing human Na+, K+-ATPase alpha1 subunit M1-M2 extracellular segment].

OBJECTIVE: To assess the binding activity of polypeptide containing human Na+, K+-ATPase alpha1 subunit M1-M2 extracellular segment (HES1 derivative). METHODS: HES1 derivative was synthesized by Fmoc method and purified by high-performance liquid chromatography-mass spectrometry, and its binding activity was identified by radioligand binding assay. RESULTS: 3H-ouabain and synthetic HES1 derivative showed some binding activity with the equilibrium dissociation constant (KD) of 24.58 nmol/L, with the the receptor density of 492.43 fmol x mg(-1) pro. and IC50 of 3.078 x 10(-7) mol/L. CONCLUSION: HES1 derivative can bind to ouabain and has the potential of becoming an effective therapeutic agent.

[Preparation and application of anti-ouabain IgY antibody].

OBJECTIVE: To prepare highly specific anti-ouabain polyclonal antibody for detecting endogenous ouabain in tissues. METHODS: Ouabain-BSA compound was used to immunize hens, and the eggs were collected one week after the first immunization. The IgY antibodies in the egg yolk were separated and purified by PEG-6000 Method, and analyzed by 12% SDS-PAGE and enzyme-linked immunosorbent assay (ELISA) for titration. The IgY antibodies obtained were applied subsequently in ELISA and immunohistochemistry. RESULTS: The IgY titer increased rapidly after the second immunization, with the highest titer of 1:10240 that lasted for at least 4 weeks. Competitive ELISA for IgY detection showed an average intraassay coefficient of variation (CV) of 2.03% and an inter-assay CV of 2.34%. Immunohistochemistry visualized the location of the endogenous ouabain mainly in the cytoplasm of the zona reticularis of rat adrenal cortex. CONCLUSION: Immunization of hens allows efficient preparation of IgY antibody which can be used in routine immunoassays.

[Comparison between anti-ouabain egg yolk(IgY) and rabbit antibody(IgG) in enzyme-linked immunosorbent assay].

AIM: To improve specificity and accuracy of endogenous ouabain measurement assay. METHODS: Anti-ouabain polyclonal antibody egg yolk (IgY) and anti-ouabain rabbit antibody (IgG) were prepared respectively. In the presence of two kinds of antibody, then the specificity and accuracy of enzyme-linked immunosorbent assay (ELISA) were compared. RESULTS: The ELISA, in the presence of IgY, provided a sensitivity of the average intraassay coefficient of variation(CV) was 2.03%, and the inter-assay CV was 2.34% respectively. In contrast, IgG were 2.83% and 3.29%. No significant interferences were observed with hydrocortisone and dexamethasone. There was 3.45% vs. 5.95%, 3.20% vs. 5.20% of crossreaction with cedilanid and digoxin. CONCLUSION: The specificity and accuracy of ELISA, in which IgY was used, were more better than IgG.