An Ultra-Sensitive Electrochemical Enzyme Immunoassay for Thyroid Stimulating Hormone in Human Serum

A sensitive heterogeneous electrochemical enzyme immunoassay has been developed for thyroid stimulating hormone (TSH) by modifying a commercially available two-site immunoenzymometric assay. p-Aminophenyl phosphate (PAPP) was used as the substrate of alkaline phosphatase, and hydrolysed to p-aminophenol (PAP). The amount of PAP produced from the assay was proportional to the amount of TSH in the sample. Detection of PAP was done by oxidative amperometry in a flow injection system. The working electrode was a glassy carbon electrode whose potential was held at +325 mV (vs Ag/AgCl). The amperometric detection of PAP required only 1 μl of sample (the range of linearity: 50.0 fmol-100 pmol PAP, the limit of detection: 10.9 fmol PAP). Intra-assay precision over the assay range of linearity (0.02–60 mIU l−1 or 0.02–60 pIU TSH) showed a maximum RSD of 8.0%, and a low detection limit of 0.01 mIU l−1 or 0.01 pIU TSH. The study also indicates that this two-site electrochemical enzyme immunoassay correlates well with the Bio-Rad\u27s immunoradiometric assay currently used in our medical center (r = 0.992, slope = 1.53, n = 43) and a highly sensitive immunochemiluminometric assay in the Nichols Institute (r = 0.986, slope = 0.499, n = 23)

Fully distributed consensus for high-order strict-feedback nonlinear multiagent systems with switched topologies

summary:This paper studies the distributed consensus problem of high-order strict-feedback nonlinear multiagent systems. By employing the adaptive backstepping technique and switched system theory, a novel protocol is proposed for MASs with switched topologies. Global information such as the number of agents and communication topology is not used. In addition, the communication topology between agents can be switched between possible topologies at any time. Based on the Lyapunov function method, the proposed adaptive protocol guarantees the complete consensus of multiagent systems without restricting the dwell time of the switched signal. Finally, two numerical examples are provided to illustrate the effectiveness and advantages of the given protocol

Clinical outcomes of S2 Alar-Iliac screw technique in the treatment of severe spinal sagittal imbalance: a retrospective 2-year follow-up study

Background: The treatment of adult spinal deformity (ASD) remains a significant challenge, especially in elderly patients. This study aimed to evaluate the outcomes of the S2AI screw technique in the treatment of severe spinal sagittal imbalance with a minimum 2-year follow-up.Methods: From January 2015 to December 2018, 23 patients with severe degenerative thoracolumbar kyphosis who underwent placement of S2AI screws for long segment fusion were retrospectively reviewed. Patients were divided into group A (no mechanical complications, 13 cases) and group B (with mechanical complications, 10 cases) according to the occurrence of mechanical complications at the last follow-up. Radiographic parameters were compared between groups preoperatively, 1 month postoperatively and at the last follow-up. Risk factors for mechanical complications were analyzed.Results: The incidence of mechanical complications was 43.5% and the revision rate was 17.4%. At 1 month postoperatively, sagittal correction was better in group A than in group B (p<0.05). The area under the curve for predicting mechanical complications of sacral slope (SS), lumbar lordosis (LL), PI (pelvic incidence)-LL at 1 month postoperatively were 0.762 (p=0.035), 0.896 (p=0.001) and 0.754 (p=0.041) respectively and the best cut-off values were 24.1°, 32.8°and 12.0°. The sagittal correction of both groups was partially lost at the last follow-up.Conclusions: A high incidence of mechanical complications was observed in long-segment corrective surgery with the S2AI screw technique for severe spinal sagittal imbalance. Inadequate sagittal correction is a risk factor for the development of mechanical complications.

Epidemiology and associations with climatic conditions of Mycoplasma pneumoniae and Chlamydophila pneumoniae infections among Chinese children hospitalized with acute respiratory infections

BACKGROUND: The incidence of severe acute respiratory tract infections in children caused by Mycoplasma pneumoniae (syn. Schizoplasma pneumoniae) and Chlamydophila pneumoniae (formerly Chlamydia pneumoniae) varies greatly from year to year and place to place around the world. This study investigated the epidemiology of M. pneumoniae and C. pneumoniae infections among children hospitalized with acute respiratory infections in Suzhou, China in the year 2006, and associations between incidence rates and climatic conditions. METHODS: Nasopharyngeal aspirates obtained from 1598 patients (aged 26.4 ± 28.3 months; range, 1 month to 13 years) were analyzed with real-time PCR and ELISA. Meteorological data were obtained from the weather bureau. RESULTS: About 18.5% of patients were infected with M. pneumoniae and, C. pneumoniae, or both. Isolated M. pneumoniae infection was positively correlated with increasing age (χ(2) = 34.76, P < 0.0001). Incidence of M. pneumoniae infection was seasonal with a peak in summer (P < 0.0001) and minimum in winter (P = 0.0001), whereas C. pneumoniae infection was low only in autumn (P = 0.02). Monthly mean temperature was strongly correlated with the incidence of M. pneumoniae infection (r = 0.825, P = 0.001). CONCLUSIONS: M. pneumoniae and C. pneumoniae are important infectious agents in hospitalized children with acute respiratory tract infections. M. pneumoniae infection showed a strong direct correlation with environmental temperature

Effects of Simvastatin on Glucose Metabolism in Mouse MIN6 Cells

The aim of this study was to investigate the effects of simvastatin on insulin secretion in mouse MIN6 cells and the possible mechanism. MIN6 cells were, respectively, treated with 0 μM, 2 μM, 5 μM, and 10 μM simvastatin for 48 h. Radio immunoassay was performed to measure the effect of simvastatin on insulin secretion in MIN6 cells. Luciferase method was used to examine the content of ATP in MIN6 cells. Real-time PCR and western blotting were performed to measure the mRNA and protein levels of inward rectifier potassium channel 6.2 (Kir6.2), voltage-dependent calcium channel 1.2 (Cav1.2), and glucose transporter-2 (GLUT2), respectively. ATP-sensitive potassium current and L-type calcium current were recorded by whole-cell patch-clamp technique. The results showed that high concentrations of simvastatin (5 μM and 10 μM) significantly reduced the synthesis and secretion of insulin compared to control groups in MIN6 cells (P<0.05). ATP content in simvastatin-treated cells was lower than in control cells (P<0.05). Compared with control group, the mRNA and protein expression of Kir6.2 increased with treatment of simvastatin (P<0.05), and mRNA and protein expression of Cav1.2 and GLUT2 decreased in response to simvastatin (P<0.05). Moreover, simvastatin increased the ATP-sensitive potassium current and reduced the L-type calcium current. These results suggest that simvastatin inhibits the synthesis and secretion of insulin through a reduction in saccharometabolism in MIN6 cells

Impact of Schistosoma japonicum Infection on Collagen-Induced Arthritis in DBA/1 Mice: A Murine Model of Human Rheumatoid Arthritis

BACKGROUND: The hygiene hypothesis suggests that helminth infections prevent a range of autoimmune diseases. METHODOLOGY/PRINCIPAL FINDINGS: To investigate the effects of S. japonicum infection on collagen-induced arthritis (CIA), male DBA/1 mice were challenged with unisexual or bisexual S. japonicum cercariae two weeks prior to bovine type II collagen (CII) immunization or at the onset of CIA. S. japonicum infection prior to CII immunization significantly reduced the severity of CIA. ELISA (enzyme linked immunosorbent assay) showed that the levels of anti-CII IgG and IgG2a were reduced in prior schistosome-infected mice, while anti-CII IgG1 was elevated. Splenocyte proliferation against both polyclonal and antigen-specific stimuli was reduced by prior schistosome infection as measured by tritiated thymidine incorporation ((3)H-TdR). Cytokine profiles and CD4(+) T cells subpopulation analysis by ELISA and flow cytometry (FCM) demonstrated that prior schistosome infection resulted in a significant down-regulation of pro-inflammatory cytokines (IFN-γ, TNF-α, IL-1β and IL-6) and Th1 cells, together with up-regulation of the anti-inflammatory cytokine IL-10 and Th2 cells. Interestingly, the expansion of Treg cells and the reduction of Th17 cells were only observed in bisexually infected mice. In addition, prior schistosome infection notably reduced the expression of pro-inflammatory cytokines and receptor activator of NF-κB ligand (RANKL) in the inflamed joint. However, the disease was exacerbated at one week after infection when established CIA mice were challenged with bisexual cercariae. CONCLUSION/SIGNIFICANCE: Our data provide direct evidence that the Th2 response evoked by prior S. japonicum infection can suppress the Th1 response and pro-inflammatory mediator and that bisexual infection with egg-laying up-regulates the Treg response and down-regulates the Th17 response, resulting in an amelioration of autoimmune arthritis. The beneficial effects might depend on the establishment of a Th2-dominant response rather than the presence of the eggs. Our results suggest that anti-inflammatory molecules from the parasite could treat autoimmune diseases

Assessing the Cumulative Contribution of New and Established Common Genetic Risk Factors to Early-Onset Prostate Cancer

We assessed the evidence for association between 23 recently reported prostate cancer (PCa) variants and early-onset PCa and the aggregate value of 63 PCa variants for predicting early-onset disease using 931 unrelated men diagnosed with PCa prior to age 56 years and 1126 male controls

Nickel oxide immobilized on the carbonized eggshell membrane for electrochemical detection of urea

Urea oxidation reaction (UOR) has been known as a viable method for renal/liver disease diagnostic detection. Here, we reported a three-dimensional (3D) nickel oxide nanoparticles dressed carbonized eggshell membrane (3D NiO/c-ESM) as a modified electrode toward urea detection. Several common physical measurements were employed to confirm its structural and morphological information. NiO/c-ESM modified electrode exhibits an outstanding performance for urea determination with a linear range from 0.05 to 2.5 mM, and limit detection of ∼20 µM (3σ). This work offered a green approach for introducing 3D nanostructure through employing biowaste ESMs as templates, providing a typical example for producing new value-added nanomaterials with urea detection

Glycyrrhizin Protects Mice Against Experimental Autoimmune Encephalomyelitis by Inhibiting High-Mobility Group Box 1 (HMGB1) Expression and Neuronal HMGB1 Release

The inflammatory mediator high-mobility group box 1 (HMGB1) plays a critical role in the pathogenesis of human multiple sclerosis (MS) and mouse experimental autoimmune encephalomyelitis (EAE). Glycyrrhizin (GL), a glycoconjugated triterpene extracted from licorice root, has the ability to inhibit the functions of HMGB1; however, GL’s function against EAE has not been thoroughly characterized to date. To determine the benefit of GL as a modulator of neuroinflammation, we used an in vivo study to examine GL’s effect on EAE along with primary cultured cortical neurons to study the GL effect on HMGB1 release. Treatment of EAE mice with GL from onset to the peak stage of disease resulted in marked attenuation of EAE severity, reduced inflammatory cell infiltration and demyelination, decreased tumor necrosis factor-alpha (TNF-α), IFN-γ, IL-17A, IL-6, and transforming growth factor-beta 1, and increased IL-4 both in serum and spinal cord homogenate. Moreover, HMGB1 levels in different body fluids were reduced, accompanied by a decrease in neuronal damage, activated astrocytes and microglia, as well as HMGB1-positive astrocytes and microglia. GL significantly reversed HMGB1 release into the medium induced by TNF-α stimulation in primary cultured cortical neurons. Taken together, the results indicate that GL has a strong neuroprotective effect on EAE mice by reducing HMGB1 expression and release and thus can be used to treat central nervous system inflammatory diseases, such as MS