A Portable Array Visualization Device Integrating Sample Preparation and Detection All-in-One for the On-Site Analysis of Complex Samples.

A gas membrane separation/array fluorescence visualization (GMS/AFV) device is developed by integrating hydrazine-based carbonized copolymer dots (PD-N2H4) for visual on-site analysis. The novel PD-N2H4 was synthesized using a "polymer template" approach, exhibiting strong blue fluorescence capable of visual sensing. The GMS/AFV device integrates sample preparation and detection all-in-one, consisting of a smartphone, a sample pretreatment system, and an optical system. In the detection procedure, the samples will be treated in the sample pretreatment system to create volatile gases. Therefore, any gas samples as well as solid and liquid samples that potentially produce volatile gases can be visually detected on-site by the device. H2S was utilized as a model analyte to test the practicality of the GMS/AFV device. The entire analysis can be finished in 3 min, and the limit of detection of H2S is as low as 3.4 µg/L. Surprisingly, the device is also capable of high-throughput sample detection, which can process 48 samples simultaneously in about 20 min. The device offers a quick, easy, cheap, and environmentally friendly way to analyze volatile gases, and it creates new opportunities for on-site detection of complex samples.

A "Polymer Template" Strategy for Carbonized Polymer Dots with Controllable Properties.

Limited avenues are available for property control of carbonized polymer dots (PDs) owing to the unsatisfactory understanding of PDs" formation. Herein, a de novo "polymer template" strategy is presented for PDs with customizable functional surface groups (FSG), size, and underlying fluorescence, with a detailed mechanism. The strategy relies on novel di-active site polymers (DASPs) prepared from alkenyl azides via [3+2] cycloaddition and guanidino hydrolysis. Benefiting from these specific reactions, the DASPs were convenient for mass production and stable for storage, and could be transformed to PDs upon addition of nucleophilic agents through nucleophilic addition and substitution at 70 °C. By regulating the types of alkenyl azides, nucleophilic agents, and reaction conditions, the as-prepare PDs could be tailored with controlled types of core, FSG, and particle size, as well as fluorescence properties of quantum yield from 8.2-55.6 %, and emission maximum from 380-500 nm. These specialties make this "polymer template" strategy a promising start for building PDs-based sensor platforms. Moreover, the strategy could further our understanding towards PDs' formation, and open up a new way to customize PDs for specific needs in the fields of analysis, catalysis, images, etc.

Safety Profile and Immunologic Responses of a Novel Vaccine Against Shigella sonnei Administered Intramuscularly, Intradermally and Intranasally: Results From Two Parallel Randomized Phase 1 Clinical Studies in Healthy Adult Volunteers in Europe.

BACKGROUND: Approximately 164,000 deaths yearly are due to shigellosis, primarily in developing countries. Thus, a safe and affordable Shigella vaccine is an important public health priority. The GSK Vaccines Institute for Global Health (GVGH) developed a candidate Shigella sonnei vaccine (1790GAHB) using the Generalized Modules for Membrane Antigens (GMMA) technology. The paper reports results of 1790GAHB Phase 1 studies in healthy European adults. METHODS: To evaluate the safety and immunogenicity profiles of 1790GAHB, we performed two parallel, phase 1, observer-blind, randomized, placebo-controlled, dose escalation studies in France ("study 1") and the United Kingdom ("study 2") between February 2014 and April 2015 (ClinicalTrials.gov, number NCT02017899 and NCT02034500, respectively) in 18-45years old subjects (50 in study 1, 52 in study 2). Increasing doses of Alhydrogel adsorbed 1790, expressed by both O Antigen (OAg) and protein quantity, or placebo were given either by intramuscular route (0.059/1, 0.29/5, 1.5/25, 2.9/50, 5.9/100µg of OAg/µg of protein; study 1) or by intradermal (ID), intranasal (IN) or intramuscular (IM) route of immunization (0.0059/0.1, 0.059/1, 0.59/10µg ID, 0.29/5, 1.2/20, 4.8/80µg IN and 0.29/5µg IM, respectively; study 2). In absence of serologic correlates of protection for Shigella sonnei, vaccine induced immunogenicity was compared to anti-LPS antibody in a population naturally exposed to S. sonnei. FINDINGS: Vaccines were well tolerated in both studies and no death or vaccine related serious adverse events were reported. In study 1, doses ≥1.5/25µg elicited serum IgG median antibody greater than median level in convalescent subjects after the first dose. No vaccine group in study 2 achieved median antibody greater than the median convalescent antibody. INTERPRETATION: Intramuscularly administered Shigella sonnei GMMA vaccine is well tolerated, up to and including 5.9/100µg and induces antibody to the OAg of at least the same magnitude of those observed following natural exposure to the pathogen. Vaccine administered by ID or IN, although well tolerated, is poorly immunogenic at the doses delivered. The data support the use of the GMMA technology for the development of intramuscular multivalent Shigella vaccines.

Human Peritoneal Mesothelial Cells Display Phagocytic and Antigen-Presenting Functions to Contribute to Intraperitoneal Immunity.

Mesothelial cells lining the peritoneal cavity are strategically positioned to respond to and counter intraperitoneal infections, cancer cells, and other challenges. We have investigated human peritoneal mesothelial cells (HPMCs) for phagocytic activity, expression of surface Major Histocompatibility Complex (MHC) class II and accessory molecules involved in antigen presentation, and the ability to present recall antigens to T cells. Phagocytosis of dextran, latex beads, and Escherichia coli was observed by flow cytometry, and internalization was visualized using confocal and electron microscopy. Flow cytometry and/or cellular enzyme-linked immunosorbent assay showed constitutive expression of ICAM-1, LFA-3, and B7-1, but not B7-2 or MHC class II. Interferon-gamma induced MHC II and ICAM-1 expression in a dose- and time-dependent manner. Importantly, HPMCs induced autologous CD3 T-lymphocyte proliferation (H incorporation) after pulse with recall antigen. Human peritoneal mesothelial cells equipped with phagocytic and antigen-presenting machinery are anticipated to have an integral role in intraperitoneal immune surveillance.

[Comparison of inter- and intra-observer reliability between GATA and SMU classification systems for spinal tuberculosis].

OBJECTIVE: To compare the inter- and intra-observer reliability of the GATA and SMU classification systems for spinal tuberculosis and assess the clinical value of SMU classification. METHODS: One hundred patients with spinal tuberculosis treated in our hospital from January 2004 to December 2011 were randomly selected for analysis, including 54 males and 46 females with a mean age of 45 years (range, 16-68 years). All the patients had X-ray, CT and MRI examinations. Five observers experienced in spinal tuberculosis independently assigned the classification using the GATA and SMU classification systems, and the assignment was repeated 3 months later to test its reproducibility. Kappa value was used to determine the intra- and inter-observer reliability. RESULTS: For GATA and SMU classification systems, the inter-observer percentage of agreement averaged (59.9∓4.84)% (κ=0.412∓0.058) and (81.6∓6.06)% (κ=0.753∓0.068), and the intra-observer percentage of agreement was (75.6∓5.27)% (κ=0.624∓0.078) and (89.8∓2.28)% (κ=0.862∓0.037), respectively. CONCLUSION: The SMU classification system of spinal tuberculosis has a higher inter-observer and intra-observer reliability than the GATA classification system, but its clinical value needs to be further tested in future clinical trials.

[Jiawei shentong zhuyu decoction prevented the occurrence of failed back surgery syndrome and its effect on serum TNF-alpha a clinical study].

OBJECTIVE: To explore the clinical roles of Jiawei Shentong Zhuyu Decoction (JSZD) in preventing the occurrence of failed back surgery syndrome (FBSS), and to observe its effect on serum tumor necrosis factor-alpha (TNF-alpha). METHODS: Totally 100 patients prepared for surgical operation due to lumbar intervertebral disc herniation were randomly assigned to the treatment group and the control group according to random number table, 50 cases in each group. Patients in the treatment group additionally took JSZD, one dose per day, taken in two portions, once in the morning and once in the evening. Those in the control group took Celecoxib Capsule (200 mg each time, once per day) and Mecobalamin Tablet (0.5 mg each time, 3 times per day). They only took Mecobalamin Tablet from the 11th day. All patients were treated for 30 days. Japanese Orthopaedic Association (JOA) score was performed before treatment, at week 1, after treatment, at 6 months of followed-ups, and at 12 months of followed-ups. And the levels of TNF-alpha in the peripheral blood were observed before treatment and at one month after treatment. RESULTS: Totally 93 patients completed the followed-up study. The JOA scores were improved after treatment, at 6 and 12 months of followed-ups (P < 0.05, P < 0.01). The JOA score at 6 months of followed-ups was superior in the treatment group to that of the control group (P < 0.05). Five patients (accounting for 10.6%) suffered from FBSS in the treatment group, while 9 (accounting for 19.6%) suffered from FBSS in the control group. The treatment group was superior to the control group (P < 0.05). The TNFalpha level was improved after treatment in the two groups. Of them, the improvement of TNF-alpha in the treatment group was better than that of the control group (P < 0.05). CONCLUSION: The application of JSZD was effective for preventing the occurrence of FBSS, and improved the serum TNF-alpha level.

Effect of vaginal immunization with HIVgp140 and HSP70 on HIV-1 replication and innate and T cell adaptive immunity in women.

The international effort to prevent HIV-1 infection by vaccination has failed to develop an effective vaccine. The aim of this vaccine trial in women was to administer by the vaginal mucosal route a vaccine consisting of HIV-1 gp140 linked to the chaperone 70-kDa heat shock protein (HSP70). The primary objective was to determine the safety of the vaccine. The secondary objective was to examine HIV-1 infectivity ex vivo and innate and adaptive immunity to HIV-1. Protocol-defined female volunteers were recruited. HIV-1 CN54gp140 linked to HSP70 was administered by the vaginal route. Significant adverse reactions were not detected. HIV-1 was significantly inhibited ex vivo in postimmunization CD4(+) T cells compared with preimmunization CD4(+) T cells. The innate antiviral restrictive factor APOBEC3G was significantly upregulated, as were CC chemokines which induce downregulation of CCR5 in CD4(+) T cells. Indeed, a significant inverse correlation between the proportion of CCR5(+) T cells and the concentration of CCL-3 or CCL-5 was found. Importantly, the upregulation of APOBEC3G showed a significant inverse correlation, whereas CCR5 exhibited a trend to correlate with inhibition of HIV-1 infection (r = 0.51). Furthermore, specific CD4(+) and CD8(+) T cell proliferative responses were significantly increased and CD4(+) T cells showed a trend to have an inverse correlation with the viral load (r = -0.60). However, HIVgp140-specific IgG or IgA antibodies were not detected. The results provide proof of concept that an innate mechanism consisting of CC chemokines, APOBEC3G, and adaptive immunity by CD4 and CD8 T cells might be involved in controlling HIV-1 infectivity following vaginal mucosal immunization in women. (This study has been registered at ClinicalTrials.gov under registration no. NCT01285141.) Importance: Vaginal immunization of women with a vaccine consisting of HIVgp140 linked to the 70-kDa heat shock protein (HSP70) elicited ex vivo significant inhibition of HIV-1 replication in postimmunization CD4(+) T cells compared with that in preimmunization peripheral blood mononuclear cells. There were no significant adverse events. The vaccine induced the significant upregulation of CC chemokines and the downmodulation of CCR5 expression in CD4(+) T cells, as well as an inverse correlation between them. Furthermore, the level of CCR5 expression was directly correlated with the viral load, consistent with the protective mechanism in which a decrease in CCR5 molecules on CD4(+) T cells decreases HIV-1 envelope binding. Expression of the antiviral restriction factor APOBEC3G was inversely correlated with the viral load, suggesting that it may inhibit intracellular HIV-1 replication. Both CD4(+) and CD8(+) T cells showed HIVgp140- and HSP70-specific proliferation. A strong inverse correlation between the proportion of CC chemokine-modulated CCR5-expressing CD4(+) T cells and the stimulation of CD4(+) or CD8(+) T cell proliferation by HIVgp140 was found, demonstrating a significant interaction between innate and adaptive immunity. This is the first clinical trial of vaginal immunization in women using only HIVgp140 and HSP70 administered by the mucosal route (3 times) in which a dual innate protective mechanism was induced and enhanced by significant adaptive CD4(+) and CD8(+) T cell proliferative responses.

[Treating cervical spinal canal stenosis by open-door cervical laminoplasty combined modified buyang huanwu decoction].

OBJECTIVE: To explore the clinical efficacy of open-door cervical laminoplasty combined Modified Buyang Huanwu Decoction (MBHD) to treat cervical spinal canal stenosis (CSCS). METHODS: Totally 32 CSCS patients were randomly assigned to two groups, Group A (17 cases, treated by laminoplasty) and Group B (15 cases, treated by laminoplasty combined MBHD). All patients received open-door cervical laminoplasty. Those in Group B took MBHD additionally for 2 weeks after surgery. The visual analogue scale (VAS), the Japanese Orthopedic Association (JOA) score, and the neck disability index (NDI) were measured preoperative, postoperative 3 months and 12 months, respectively. RESULTS: There was no statistical difference in preoperative VAS, JOA, or NDI (P > 0.05). The VAS, JOA, and NDI were obviously improved 3 months and 12 months after surgery in the two groups, showing statistical difference when compared with before surgery in the same group (P < 0.01). At 3 months after surgery the aforesaid indices in Group B were superior to those in Group A (P < 0.05). There was no statistical difference in the aforesaid indices between the two groups at 12 months after surgery (P > 0.05). CONCLUSION: MBHD favorably improved early recovery of neural functions of CSCS patients (3 months after surgery).

[Minimally invasive surgical treatment for lumbar degenerative disease with IsoC-3D navigation under Mast Quadrant system].

OBJECTIVE: To evaluate the effects of surgical treatment of IsoC-3D navigation assisted percutaneous pedicle screw fixation,vertebral canal decompression and interbody fusion using Mast Quadrant system in patients with lumbar degenerative disease. METHODS: From January 2009 to February 2010,21 patients with lumbar degenerative disease were treated with IsoC-3D navigation under Mast Quadrant system. There were 12 males and 9 females, with an average age of 50.2 years (ranged,36 to 72 years). All patients underwent discectomy,vertebral canal decompression, cage implantation using Mast Quadrant system and IsoC-3D navigation assisted sextant lumbar fixation. Clinical outcomes were evaluated with Oswestry disability index (ODI), Visual analog scale (VAS) and degree of satisfaction of patients. RESULTS: Eighteen patients (85.7%) were followed up from 6 to 18 months with an average of 10 months. No surgery-related complications were found. The preoperative, postoperative ODI scores was 49.6 +/- 12.2 and 17.2 +/- 9.2, respectively (P < 0.01); VAS score of leg pain decreased from preoperative 75.2 +/- 10.0 to 12.2 +/- 11.8 at final follow-up (P < 0.01); VAS score of lumbago decreased from preoperative 59.9 +/- 17.3 to 16.6 +/- 11.5 at final follow-up (P < 0.01). Sixteen patients obtained satisfactory results. CONCLUSION: IsoC-3D navigation assisted percutaneous pedicle screw fixation,vertebral canal decompression and interbody fusion using Mast Quadrant system could achieve satisfactory clinical results in treating lumbar degenerative disease and may be a better alternative to conventional surgical procedures. It has advantages such as limited tissue damage, less blood loss, short time in hospital.

Systemic and mucosal immune responses to sublingual or intramuscular human papilloma virus antigens in healthy female volunteers.

UNLABELLED: The sublingual route has been proposed as a needle-free option to induce systemic and mucosal immune protection against viral infections. In a translational study of systemic and mucosal humoral immune responses to sublingual or systemically administered viral antigens, eighteen healthy female volunteers aged 19-31 years received three immunizations with a quadravalent Human Papilloma Virus vaccine at 0, 4 and 16 weeks as sublingual drops (SL, n = 12) or intramuscular injection (IM, n = 6). IM antigen delivery induced or boosted HPV-specific serum IgG and pseudovirus-neutralizing antibodies, HPV-specific cervical and vaginal IgG, and elicited circulating IgG and IgA antibody secreting cells. SL antigens induced ~38-fold lower serum and ~2-fold lower cervical/vaginal IgG than IM delivery, and induced or boosted serum virus neutralizing antibody in only 3/12 subjects. Neither route reproducibly induced HPV-specific mucosal IgA. Alternative delivery systems and adjuvants will be required to enhance and evaluate immune responses following sublingual immunization in humans. TRIAL REGISTRATION: ClinicalTrials.govNCT00949572.

An investigation of clinical and immunological events following repeated aerodigestive tract challenge infections with live Mycobacterium bovis Bacille Calmette Guérin.

Bacille Calmette Guérin substrain Moreau Rio de Janeiro is an attenuated strain of Mycobacterium bovis that has been used extensively as an oral tuberculosis vaccine. We assessed its potential as a challenge model to study clinical and immunological events following repeated mycobacterial gut infection. Seven individuals received three oral challenges with approximately 10(7) viable bacilli. Clinical symptoms, T-cell responses and gene expression patterns in peripheral blood were monitored. Clinical symptoms were relatively mild and declined following each oral challenge. Delayed T-cell responses were observed, and limited differential gene expression detected by microarrays. Oral challenge with BCG Moreau Rio de Janeiro vaccine was immunogenic in healthy volunteers, limiting its potential to explore clinical innate immune responses, but with low reactogenicity.

Transient facial nerve paralysis (Bell's palsy) following intranasal delivery of a genetically detoxified mutant of Escherichia coli heat labile toxin.

BACKGROUND: An association was previously established between facial nerve paralysis (Bell's palsy) and intranasal administration of an inactivated influenza virosome vaccine containing an enzymatically active Escherichia coli Heat Labile Toxin (LT) adjuvant. The individual component(s) responsible for paralysis were not identified, and the vaccine was withdrawn. METHODOLOGY/PRINCIPAL FINDINGS: Subjects participating in two contemporaneous non-randomized Phase 1 clinical trials of nasal subunit vaccines against Human Immunodeficiency Virus and tuberculosis, both of which employed an enzymatically inactive non-toxic mutant LT adjuvant (LTK63), underwent active follow-up for adverse events using diary-cards and clinical examination. Two healthy subjects experienced transient peripheral facial nerve palsies 44 and 60 days after passive nasal instillation of LTK63, possibly a result of retrograde axonal transport after neuronal ganglioside binding or an inflammatory immune response, but without exaggerated immune responses to LTK63. CONCLUSIONS/SIGNIFICANCE: While the unique anatomical predisposition of the facial nerve to compression suggests nasal delivery of neuronal-binding LT-derived adjuvants is inadvisable, their continued investigation as topical or mucosal adjuvants and antigens appears warranted on the basis of longstanding safety via oral, percutaneous, and other mucosal routes.

Induction of protective serum meningococcal bactericidal and diphtheria-neutralizing antibodies and mucosal immunoglobulin A in volunteers by nasal insufflations of the Neisseria meningitidis serogroup C polysaccharide-CRM197 conjugate vaccine mixed with chitosan.

Thirty-six healthy volunteers received either a single intramuscular injection of Neisseria meningitidis serogroup C polysaccharide (MCP)-CRM197 conjugate vaccine in alum or two nasal insufflations 28 days apart of the same vaccine powder, without alum, mixed with chitosan. Nasal immunization was well tolerated, with fewer symptoms reported than after intramuscular injection. The geometric mean concentrations of MCP-specific immunoglobulin G (IgG) after one nasal immunization were 3.25 microg/ml in naïve subjects and 14.4 microg/ml in subjects previously immunized parenterally, compared with 4.30 microg/ml in naïve subjects immunized intramuscularly. The geometric mean titer of serum bactericidal antibody (SBA) rose 24-fold after two nasal immunizations in naïve subjects and was comparable to parenteral immunization (1,080 versus 1,625). All subjects achieved SBA titers associated with protection after two nasal immunizations: even those with titers of <8 at entry. A single nasal immunization boosted the SBA titer to > or =128 in 96% of previously immunized subjects, and two immunizations achieved this level in 92% of naive subjects. MCP-specific IgG levels were approximately 70% IgG2 and approximately 20% IgG1 after nasal or intramuscular immunization. Increases in CRM197-specific IgG and diphtheria toxin-neutralizing activity were observed after nasal or intramuscular immunization, with balanced IgG1/IgG2 and higher IgG4. Significant MCP-specific secretory IgA was detected in nasal wash only after nasal immunization and predominantly on the immunized side. Simple nasal insufflation of existing MCP-CRM197 conjugate vaccines in chitosan offers an inexpensive but effective needle-free prime and boost against serogroup C N. meningitidis and diphtheria.