Rationally Designed Self-Assembling Nanovaccines Elicit Robust Mucosal and Systemic Immunity against Rhabdovirus.

Viral diseases have constantly caused great threats to global public health, resulting in an urgent need for effective vaccines. However, the current viral vaccines often show low immunogenicity. To counter this, we report a smart strategy of a well-designed modular nanoparticle (LSG-TDH) that recapitulates the dominant antigen SG, low-molecular-weight protamine, and tetralysine-modified H-chain apoferritin (TDH). The constructed LSG-TDH nanovaccine could self-assemble into a nanocage structure, which confers excellent mucus-penetrating, cellular affinity, and uptake ability. Studies demonstrate that the LSG-TDH nanovaccine could strongly activate both mucosal and systemic immune responses. Importantly, by immunizing wild-type and TLR2 knockout (TLR2-KO) zebrafish, we found that TLR2 could mediate LSG-TDH-induced adaptive mucosal and systemic immune responses by activating antigen-presenting cells. Collectively, our findings offer new insights into rational viral vaccine design and provide additional evidence of the vital role of TLR2 in regulating adaptive immunity.

Study on neutron energy spectrum unfolding algorithm with EJ309 liquid scintillation detector.

Proton recoil method can be used to experimentally measure fast neutron energy spectrum of non-pulsed neutron sources. The neutron energy spectrum unfolding algorithms based on the MLEM method, the GOLD deconvolution method, the Direct-D method, have been developed by using the EJ309 liquid scintillation detector. The degree of iteration by the mean square error (MSE) is proposed as a judgment criterion by according to the iterative accuracy, convergence speed and iteration efficiency. The developed neutron energy spectrum unfolding algorithms can unfolding the standard simulated mono-energetic neutron spectrum (2.5 MeV), 252Cf neutron spectrum, Am-Be neutron spectrum and the experimentally measured D-D neutron spectrum with higher precision as well as fewer iterations. The unfolded neutron spectra are in good agreement with the standard simulated neutron spectra and evaluated D-D neutron spectrum, which is revealed that the developed unfolding algorithms can unfolding neutron energy spectrum with reasonable accuracy.

Dual-Targeting Polymer Nanoparticles Efficiently Deliver DNA Vaccine and Induce Robust Prophylactic Immunity against Spring Viremia of Carp Virus Infection.

Spring viremia of carp virus (SVCV) is highly contagious and lethal to most cyprinid fish, causing serious economic losses to the carp aquaculture industry. Although DNA vaccines can generate long-term humoral and cellular immune responses, which provide protective immunity against SVCV, the major drawback of DNA vaccines is their low immunogenicity in clinical tests. Here, we construct a dual-targeted polymer DNA vaccine delivery platform (MCS-PCHG) by using mannosylated chitosan to encapsulate the poly(d,l-lactide-co-glycolide)-loaded DNA vaccine containing the heavy-chain CH3 region (CH3) of common carp IgM and the antigenic domain (G131c). The developed nanovaccine delivery platform showed good biocompatibility in vivo and in vitro. With the modification of the mannose moiety and the modification of CH3, the constructed MCS-PCHG could efficiently activate the maturation of antigen-presenting cells. Moreover, we observe significantly high level of immune-related genes expression, serum antigen-specific IgM, SVCV-neutralizing antibody titers in fish vaccinated with MCS-PCHG. Next, the protective efficacy of MCS-PCHG was further evaluated by challenge test. The highest survival rate (ca. 84%) was observed in fish vaccinated with MCS-PCHG after challenging with SVCV. This study presents a novel design for smart, dual-targeted polymer nanoparticles, which are inherently biocompatible, promising for targeted vaccine delivery. IMPORTANCE Spring viremia of carp virus (SVCV) affects global cyprinid fish farming industry, with no available commercial vaccine. Herein, we developed a dual-targeting polymer nanovaccine (MCS-PCHG) by using mannose and common carp IgM heavy chain CH3 region (CH3) as antigen presenting cell (APCs) recognition moiety, attaining the effective delivery of antigen. This dual-targeting polymer vaccine can efficiently activate the APCs, and further induce robust and durable adaptive immune response with good protection against SVCV infection. Our study provides valuable theoretical basis for developing efficient vaccine against infectious diseases in aquaculture.

TLR2-mediated mucosal immune priming boosts anti-rhabdoviral immunity in early vertebrates.

Most pathogens utilize mucosal surfaces to enter and propagate within the host. As one of the main pathogens of fatal and highly contagious diseases, rhabdoviruses are distributed widely in nature affecting both human and animals. Therefore, local mucosal immune responses, most effectively induced by mucosal vaccines, act as frontline immunity to block the pathogens at its initial replication sites. However, the underlying regulatory mechanisms of mucosal immunity triggered by mucosal vaccine remains unclear. Herein, a rhabdoviruses glycoprotein-based mucosal vaccine (G131c) was used to elucidate the regulatory mechanism of local mucosal immunity in zebrafish, a typical immunological model. Firstly, we verified the strong immunoprotection of G131c mucosal vaccine. Furthermore, the delivery kinetics of G131c was evaluated in vivo, indicating the effective uptake of vaccines by mucosal tissues through immersion vaccination. Importantly, we demonstrate immersion with G131c vaccine could activate antigen presenting cells (APCs) at the local mucosal sites, and then arose robust local mucosal and systemic immune responses. More critically, we found that G131c mediated these immune effects by interacting with Toll-like receptor 2 (TLR2) and activating downstream MAPK and NF-κB signaling pathways. Thus, our findings provide previously unappreciated evidence that rhabdovirus glycoprotein could interact with TLR2 and then activate the APCs in local mucosal sites. This study provides a comprehensive perspective on the mechanism of TLR2-mediated mucosal immunity in the early vertebrates.

Immersion immunization of common carp with bacterial ghost-based DNA vaccine inducing prophylactic protective immunity against spring viraemia of carp virus.

The interactive applications of immunization route, vaccine type and delivery vectors are emerging as a key area of research within the field of mass immunization in fishery production. In an effort to improve DNA vaccine's immune efficiency in large-scale immunization, a promising bacterial ghost-loaded DNA vaccine was constructed based on Escherichia coli DH5α. In common carp was investigated the immune response to immersion immunization via related indicator analysis, and the challenge test of spring viraemia of carp virus (SVCV) was carried out. The result indicated that BG-loaded DNA vaccine induced higher serum antibody level than naked pEG-G. Simultaneously, the immunophysiological indicators and genes change at the more advanced levels in the BG/pEG-G immune group. At the treatment concentration of 20 mg/L of the BG/pEG-G group, IgM and IgZ expressions in vivo were markedly increased by 21.62 times and 6.91 times, respectively, and the relative percentage survival reached the peak of 59.57%. This study paves the way for future aquatic animal vaccine research, which aimed to develop the highly effective immersion vaccine system by delivery vectors, with the ultimate aim to prevent and restrict SVCV in actual production.

Catalytic conversion of corncob to furfuryl alcohol in tandem reaction with tin-loaded sulfonated zeolite and NADPH-dependent reductase biocatalyst.

In this study, tin-loaded sulfonated zeolite (Sn-zeolite) catalyst was synthesized for catalysis of raw corncob (75.0 g/L) to 103.0 mM furfural at 52.3% yield in water (pH 1.0) at 170 °C. This corncob-derived furfural was subsequently biotransformed with recombinant E. coli CG-19 cells coexpressing NADPH-dependent reductase and glucose dehydrogenase at 35 °C by supplementary of glucose (1.5 mol glucose/mol furfural), sodium dodecyl sulfate (0.50 mM) and NADP+ (1.0 µmol NADP+/mmol furfural) in the aqueous catalytic media (pH 7.5). Both sodium dodecyl sulfate (0.50 mM) and Sn4+ (1.0 mM) could promote reductase activity by 1.4-folds. Within 3 h, furfural was wholly catalyzed into furfuryl alcohol. By combining chemical catalysis with Sn-zeolite and biocatalysis with CG-19 cells in one-pot, an effective and sustainable process was established for tandemly catalyzing renewable biomass into furfuryl alcohol under environmentally-friendly way.

Application of Biomimetic Cell-Derived Nanoparticles with Mannose Modification as a Novel Vaccine Delivery Platform against Teleost Fish Viral Disease.

Although cell membrane-coated nanoparticles are widely used as a promising nanodelivery platform, a few studies reported their application in developing the teleost nanovaccine delivery system. Here, we present a biomimetic vaccine delivery platform by encapsulating chitosan-loaded DNA vaccine with teleost erythrocytes membrane modified by mannose. The developed CS-G@M-M nanovaccine delivery platform shows good biocompatibility in vivo and in vitro. With further modification of mannose moiety, the constructed CS-G@M-M showed enhanced uptake by antigen-presenting cells (APCs) and increased accumulation of CS-G@M-M in immune tissues including spleen, kidney, and hindgut. Critically, using a quantitative real-time polymerase chain reaction (qRT-PCR) assay, increased mRNA levels of immune-related genes were detected in spleen and hindgut of vaccinated fish. Moreover, through enzyme-linked immunosorbent assay (ELISA), we found that the levels of CD80/86, TNF-α, IgM, and IgZ in spleen and hindgut were significantly increased. To evaluate the immunoprotection efficacy of the constructed nanovaccine, spring viremia of carp virus (SVCV), a rhabdovirus of worldwide importance that requires notification within 48 h to the International Office of Epizootics once detected, was used as a model for virus challenge. We carried out three challenge tests on 3rd, 21st, and 70th days post vaccination, respectively. Notably, CS-G@M-M nanovaccine showed durability of immunoprotection efficacy that could protect zebrafish from SVCV challenge. This work presents a novel design of smart teleost erythrocytes membrane-coated nanoparticles, which are inherently biocompatible, promising for eliciting robust adaptive immune responses in preventing fish viral diseases.

Distinct roles of neuregulin in different models of neuropathic pain.

The objective of the study was to investigate the role of neuregulin-ErbB signaling in neuropathic pain in different types of injury. Neuregulin-1(NRG-1) was injected into animals with either formalin-induced pain model or spared nerve injury (SNI) model. Formalin tests or paw withdrawal tests were performed to study the role of NRG-1 in neuropathic pain. siRNA specific to different erbB receptors were then introduced to test which specific signaling pathway was required for NRG-1 signaling in the different pain models. NRG-1 inhibits neuropathic pain after SNI in a dose-dependent manner, while NRG-1 aggravates formalin-induced neuropathic pain. ErbB2 and erbB4 receptors were activated after neuregulin administration. Knockdown of ErbB2 relieves the aggravation of NRG-1 on formalin-induced neuropathic pain, and knockdown of ErbB4 could relieve the inhibition of NRG-1 on neuropathic pain in the SNI model. NRG-1 has two distinct functions depending on the different receptor activation in different models of neuropathic pain. These novel findings may provide new therapeutic approaches for the treatment of neuropathic pain in different injury types.

miR-210 activates notch signaling pathway in angiogenesis induced by cerebral ischemia.

The compensatory angiogenesis that occurs after cerebral ischemia increases blood flow to the injured area and limits extension of the ischemic penumbra. In this way, it improves the local blood supply. Fostering compensatory angiogenesis is an effective treatment for ischemic cerebrovascular disease. However, angiogenesis in the adult organism is a complex, multi-step process, and the mechanisms underlying the regulation of angiogenesis are not well understood. Although Notch signaling reportedly regulates the vascularization process that occurs in ischemic tissues, little is known about the role of Notch signaling in the regulation of ischemia-induced angiogenesis after ischemic stroke. Recent research has indicated that miR-210, a hypoxia-induced microRNA, plays a crucial role in regulating the biological processes that occur in blood vessel endothelial cells under hypoxic conditions. This study was undertaken to investigate the role of miR-210 in regulating angiogenesis in response to brain ischemia injury and the role of the Notch pathway in the body's response. We found miR-210 to be significantly up-regulated in adult rat ischemic brain cortexes in which the expression of Notch1 signaling molecules was also increased. Hypoxic models of human umbilical vein endothelial cells (HUVE-12) were used to assess changes in miR-210 and Notch1 expression in endothelial cells. Results were consistent with in vivo findings. To determine the molecular mechanisms behind these phenomena, we transfected HUVE-12 cells with miR-210 recombinant lentiviral vectors. We found that miR-210 overexpression caused up-regulation of Notch1 signaling molecules and induced endothelial cells to migrate and form capillary-like structures on Matrigel. These data suggest that miR-210 is involved in the regulation of angiogenesis in response to ischemic injury to the brain. Up-regulation of miR-210 can activate the Notch signaling pathway, which may contribute to angiogenesis after cerebral ischemia.

A prospective clinical study of routine repeat computed tomography (CT) after traumatic brain injury (TBI).

PURPOSE: To discuss the repeated CT scanning in patients with traumatic brain injury (TBI) and to identify the conditions under which this approach is necessary. METHODS: One hundred and seventy-one patients who suffered TBI but were not surgically treated were divided into two groups: the routine-repeat CT group (n = 89) and the non-routine-repeat CT group (n = 82). The patients' clinical characteristics were compared. T-tests and stepwise logistic regression were used for analysis. Patients in the routine-repeat CT group were divided into three groups according to GCS scores to determine the need for routinely repeated CT scans. RESULTS: The results revealed statistically significant differences between the two groups in terms of neuro-ICU-LOS and LOS (p < 0.01). No significant differences emerged with respect to hospital charges and GCS scores at discharge (p > 0.05). AGE, international normalized ratio (INR), D-dimer concentration (DD), GCS scores and number of hours between the first CT scan and the injury (HCT1) were influential factors of developing progressive haemorrhage. CONCLUSION: The routine-repeat CT group fared better than did the non-routine-repeat CT group. Routinely repeated CTs were minimally effective among those with mild TBI, whereas this procedure demonstrated a significant effect on patients with moderate and severe TBI.