High expression of the classical swine fever virus (CSFV) envelope protein E2 by a single amino acid mutation and its embedded in the pseudorabies virus (PRV) vector for immunization.

Pseudorabies (PR) and classical swine fever (CSF) are economically important infectious diseases in pigs. Most pig farms in China are vaccinated against these two diseases. Gene-deleted pseudorabies virus (PRV) can be used to develop promising and economical multivalent live attenuated viral vector vaccines. It has been reported that recombinant PRV can express a truncated E2 protein (1-338 aa), but it has not been reported that recombinant PRV can express a full-length E2 protein. We constructed nine groups of E2 proteins with different expression forms and found that the E2 protein could be expressed in vitro only when the transmembrane region of E2 was removed and the signal peptide was added. Analysis of the transmembrane region of E2 revealed that the high hydrophobicity of the E2 transmembrane region was the main reason for its inability to express. By mutating an amino acid to reduce the hydrophobicity of the transmembrane region, it was found that the full-length mutant of E2 (E2FL-muta3 or E2FL-muta4) could be expressed. The expressed full-length mutant E2 could also localize to the cell membrane. Mice immunized with a PRV vector vaccine expressing E2FL-muta3 or E2FL-muta4 developed specific cellular immunity to the E2 protein and stimulated higher levels of E2 antibody than mice immunized with a PRV vector expressing truncated E2. After immunizing the rabbits, the lethal challenge by PRV-ZJ2013 and the febrile response elicited by CSFV were simultaneously prevented. These results suggest that rPRV-dTK/gE-E2FL-muta4 is a promising bivalent vaccine against CSFV and PRV infections.

Recombinant pseudorabies virus (PRV) expressing stabilized E2 of classical swine fever virus (CSFV) protects against both PRV and CSFV.

Pseudorabies (PR) and classical swine fever (CSF) are economically important infectious diseases of pigs. Most pig farms in China are immunized against these two diseases. Here, we describe a stabilized E2 protein as an immunogen inserted into the PRV genome as a bivalent live virus-vectored vaccine. The E2 protein has 48 variant sites, there are 2-5 candidate amino acids per variant site, and the relative energy contribution of each amino acid to E2 energy was calculated. Combined substitutions of amino acids at the neighbor variant site (neighbor substitution) were performed to obtain the E2 protein sequence with the lowest energy (stabilized E2). Multiple amino acid substitutions at 48 variant sites were performed, and the results were consistent with neighbor substitutions. The stabilized E2 sequence was obtained, and its energy decreased by 22 Rosetta Energy Units (REUs) compared with the original sequence. After the recombinant PRV expressing stabilized E2 of CSFV was constructed, the secretion efficiency of stabilized E2 was increased by 2.97 times, and the thermal stability was increased by 10.5 times. Immunization of mice resulted in a 2-fold increase in antibody production, and a balanced antibody level against subtype 1.1 and subtype 2.1d E2 was achieved. In rabbits immunized, the lethal challenge of PRV-ZJ and the fever response induced by CSFV could be prevented simultaneously. These findings suggest that rPRV-muta/287aaE2 is a promising bivalent vaccine against CSFV and PRV infections.

Effects of High-Intensity Strength Training in Adults With Knee Osteoarthritis: A Systematic Review and Meta-analysis of Randomized Controlled Trials.

OBJECTIVE: The aim of the study is to assess the effects of high-intensity strength training versus low-intensity strength training or routine care in adults with knee osteoarthritis. DESIGN: PubMed, Embase, Cochrane Library, and Web of Science were searched up to March 10, 2021. The outcomes were knee pain, knee function, quality of life, and adverse events. RESULTS: Ten studies of 892 subjects with knee osteoarthritis were included. No significant differences were found between the high-intensity strength training group and the low-intensity strength training or routine care group in the Western Ontario and McMaster Universities Osteoarthritis Index pain score, Knee Injury and Osteoarthritis Outcome Score pain score, Western Ontario and McMaster Universities Osteoarthritis Index stiffness score, Western Ontario and McMaster Universities Osteoarthritis Index physical function score, Knee Injury and Osteoarthritis Outcome Score symptom score, Knee Injury and Osteoarthritis Outcome Score activities of daily living score, Knee Injury and Osteoarthritis Outcome Score sport and recreation score, Timed Up and Go result, gait velocity, walking time, peak torque of the knee extensors, Knee Injury and Osteoarthritis Outcome Score quality of life score, and adverse event incidence (all P > 0.05). The peak torque of the knee flexors at 120-degree per sec contraction (pooled weighted mean difference, 7.520; 95% confidence interval, 1.256 to 13.784; P = 0.019) in the high-intensity group was improved significantly than that in the low-intensity training or routine care group. CONCLUSIONS: High-intensity strength training may have similar effects in improving knee pain, knee function, and quality of life, with comparable safety to low-intensity strength training and routine care.

TRIM59 attenuates IL-1ß-driven cartilage matrix degradation in osteoarthritis via direct suppression of NF-κB and JAK2/STAT3 signaling pathway.

The tripartite motif (TRIM) protein family are implicated in a wide array of cellular processes, including cell growth, differentiation, apoptosis and inflammation. This study aimed to investigate the specific function of TRIM59 in chondrocytes and its association with the pathophysiology of osteoarthritis (OA). We observed the downregulated TRIM59 expression in OA cartilage compared to normal tissues. Overexpression of TRIM59 suppressed interleukin 1 beta (IL-1ß)-induced extracellular matrix (ECM) metabolic imbalance, proinflammatory cytokine production, apoptosis and decrease in cell viability. Mechanistic analyses further revealed that IL-1ß-induced activation of the NF-κB and JAK2/STAT3 pathway is suppressed upon TRIM59 overexpression. TRIM59 expression was consistently decreased in a rat OA model in vivo, and its overexpression led to inhibition of matrix metallopeptidase-13 (MMP-13) production, proinflammatory cytokine levels and increased collagen type II (collagen II) and aggrecan synthesis. Our data collectively suggest that TRIM59 plays a critical in OA development through regulation of NF-κB and JAK2/STAT3 signaling pathway. Pharmacological upregulation of TRIM59 may therefore present an effective novel therapeutic approach for OA.

Peptidylarginine deiminase 4 -104C/T polymorphism and risk of rheumatoid arthritis: A pooled analysis based on different populations.

BACKGROUND: Many studies have analyzed the association between peptidylarginine deiminase 4 (PADI4) -104C/T polymorphism and rheumatoid arthritis (RA). However, the results are inconsistent. This meta-analysis, based on different populations, updated and reevaluated the possible associations between PADI4 -104C/T polymorphism and the susceptibility to RA. METHODS: A literature search was performed on PubMed and related Chinese databases up to April 2017. The association between PADI4 -104C/T polymorphism and RA risk was evaluated by calculating pooled odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: A total of seventeen studies, including 5,756 RA cases and 4,987 controls, were screened out. In the overall population, PADI -104C/T polymorphism was significantly associated with an increased RA risk. In this meta-analysis stratified by ethnicity, a significant association between PADI -104C/T polymorphism and RA risk was established in China and Japan. CONCLUSIONS: Our study indicated a significantly increased association between PADI -104C/T polymorphism and RA in Chinese and Japanese populations. Because most included populations in this meta-analysis were Asian, further studies are needed to elucidate whether the PADI4 -104C/T gene confers RA in other ethnic groups.

Self-gauged fiber-optic micro-heater with an operation temperature above 1000°C.

We report a fiber-optic micro-heater based on a miniature crystalline silicon Fabry-Perot interferometer (FPI) fusion spliced to the endface of a single-mode fiber. The silicon FPI, having a diameter of 100 µm and a length of 10 or 200 µm, is heated by a 980 nm laser diode guided through the lead-in fiber, leading to a localized hot spot with a temperature that can be conveniently tuned from the ambient temperature to >1000°C in air. In the meantime, using a white light system operating in the 1550 nm wavelength window where the silicon is transparent, the silicon FPI itself also serves as a thermometer with high resolution and high speed for convenient monitoring and precise control of the heater temperature. Due to its small size, high temperature capability, and easy operation, the micro-heater is attractive for applications in a variety of fields, such as biology, microfluidics system, mechanical engineering, and high-temperature optical sensing. As an example, the application of this micro-heater as a micro-boiler and micro-bubble generator has been demonstrated.

[Studies on the apoptosis of RK13 cells induced by rabbit hemorrhagic disease virus].

The apoptosis of RK13 cells induced by RHDV was investigated with DAPI staining, DNA ladder, Caspase 3 activity and flow cytometry, etc. The results showed that nuclear staining of infected cells with DAPI showed gradually morphological changes of the nuclei. As shown in the paper, a canonic oligonucleosome-sized DNA ladder was observed in cells harvested at 24h, 48h and 72h post-infection, confirming that DNA fragmentation was induced by RHDV infection. The results of flow cytometry showed that about 63% of cells were in apoptosis at 48h post-infection. Besides, we also demonstrated that the activation of Caspase 3 occurred during the infection process. In conclusion, our results showed that apoptosis in RHD might be determinant in the development of the pathogenesis of RHD.

[Construction of rabbit hemorrhagic disease virus replicons and its replication in RK-13 cells].

To provide an efficient and safe technology platform for studying the replication and pathogenesis mechanisms of RHDV, the interaction between the RHDV and its host cells, a replicon system of RHDV, was constructed based on the infectious cDNA clone of RHDV, in which VP60 gene encoding the capsid protein was deleted, but all the necessary protease coding regions and non-coding regions were retained. Results from RT-PCR, IFA and qRT-PCR confirmed that the replicon RNA could efficiently replicate in RK-13 cells. Besides, the results also suggested that the capsid protein which is the structural protein of RHDV is necessary for maintaining the viral infectivity.

Subwavelength focusing and guiding of surface plasmons.

The constructive interference of surface plasmon polaritons (SPP) launched by nanometric holes allows us to focus SPP into a spot of high near-field intensity having subwavelength width. Near-field scanning optical microscopy is used to map the local SPP intensity. The resulting SPP patterns and their polarization dependence are accurately described in model calculations based on a dipolar model for the SPP emission at each hole. Furthermore, we show that the high SPP intensity in the focal spot can be launched and propagated on a Ag strip guide with a 250 x 50 nm2 cross section, thus overcoming the diffraction limit of conventional optics. The combination of focusing arrays and nano-waveguides may serve as a basic element in planar nano-photonic circuits.