Objective:To evaluate the effect of hydrogen on the expression of hippocampal cold-inducible RNA-binding protein (CIRP) after cardiac arrest-resuscitation in rats.Methods:Ninety clean-grade healthy male Sprague-Dawley rats, weighing 280-320 g, were randomly divided into 3 groups: sham group (group Sham, n=20), cardiac arrest-cardiopulmonary resuscitation group (group CPR, n=35), and hydrogen-rich saline group (group H 2, n=35). Cardiac arrest was induced by transoesophageal cardiac pacing followed by CPR in group CPR.Only femoral arteriovenous puncture and tracheal intubation were performed in group Sham.Hydrogen-rich saline 5 ml/kg was intraperitoneally injected immediately after recovery of spontaneous circulation (ROSC) and at 6 and 12 h after ROSC in group H 2 , while the equal volume of normal saline was given instead in the other two groups.Neuro-functional deficit was assessed using neurologic deficit scores (NDS) at 1 and 3 days after ROSC.The animals were sacrificed immediately after intubation in group Sham and at 6 h and 1, 2 and 3 days after ROSC in CPR and H 2 groups, and the hippocampal tissues were obtained to detect the expression of nuclear and cytoplasmic CIRP by Western blot. Results:Compared with group Sham, NDS was significantly decreased at each time point after ROSC in group CPR and group H 2, the expression of nuclear CIRP was significantly down-regulated at 1, 2 and 3 days after ROSC, and the expression of cytoplasmic CIRP was up-regulated at 1 and 2 days after ROSC in group CPR, and the expression of nuclear CIRP was significantly down-regulated at each time point after ROSC, and the expression of cytoplasmic CIRP was down-regulated at 2 and 3 days after ROSC in group H 2 ( P<0.05). Compared with group CPR, NDS was significantly increased at each time point after ROSC, the expression of nuclear CIRP was down-regulated at 6 h after ROSC, and the expression of cytoplasmic CIRP was down-regulated at 1 and 2 days after ROSC in group H 2 ( P<0.05). Conclusion:The nechanism by which hydrogen reduces brain injury after cardiac arrest-resuscitation may be related to down-regulating hippocampal CIRP expression in rats.

Objective:To reduce the immunogenicity of vaccinia virus vector by replacing the D8L region, which is a neutralizing antibody epitope in vaccinia virus, with an exogenous gene.Methods:A gene fragment encoding influenza virus hemagglutinin (HA) was inserted into the D8L region to replace it using homologous recombination technique. Then, a recombinant vaccinia virus influenza vaccine was constricted. A recombinant vaccinia virus vaccine with the TK region expressing HA was used as a control. The expression of HA was validated by Western blot. BALB/c mice were immunized with the vaccines and the serum antibody titers two weeks after each immunization were evaluated by ELISA and hemagglutination inhibition assay. The protective efficacy of the recombinant vaccinia virus was assessed through a challenge experiment.Results:Western blot confirmed the successful expression of HAD8L protein in the constructed recombinant vaccines. ELISA and hemagglutination inhibition assay showed that after the primary immunization, the anti-HA antibody titer induced by the recombinant vaccinia virus with D8L region mutation was slightly higher than that induced by the vaccine with TK region mutation, and the difference was statistically significant with the increase of immunization times ( P<0.05). The recombinant vaccinia virus with D8L region mutation showed significantly lower immunogenicity than the recombinant virus with TK region mutation after the primary immunization, but there was no significant difference between them with the increase of immunization times ( P>0.05). After H1N1pdm challenge, no virus was detected in the mice immunized with the recombinant vaccinia virus with D8L region mutation and the mice showed mild lung inflammation and less tissue damage. Conclusions:This study indicated that inserting exogenous genes into the D8L region of the neutralizing antibody epitope in the vaccinia virus vector could help to reduce the immunogenicity of the vector itself and enhance the immunogenicity of the exogenous genes. This provided a reference for the use of the vaccinia virus vector as a delivery tool in the field of vaccines or gene therapy.

OBJECTIVE: To explore the effects and mechanisms of the long-snake moxibustion on ankylosing spondylitis (AS) based on Th17/Treg/Th1 immune imbalance. METHODS: A total of 60 AS patients were randomized into an observation group and a control group, 30 cases in each one. In the observation group, the long-snake moxibustion therapy was used on the acupoints of the governor vessel from Dazhui (GV 14) to Yaoshu (GV 2) as well as the bilateral Jiaji (EX-B 2) alternatively. The moxibustion was given once a day, for 7 days continuously as one course. There were 3 days at the interval between the courses and 4 courses were required. In the control group, the routine western medication was provided, the salazosulfapyridine combined with non-steroidal anti-inflammatory drugs were used, for 7 days continuous as one course. A total of 4 courses of medication were required. The enzyme linked immunosorbent assay (ELISA) was adopted to determine the levels of interleukin-6 (IL-6), interleukin-17 (IL-17), interleukin-23 (IL-23) and tumor necrosis factor-α (TNF-α). The real-time quantification polymerase chain reaction (RT-PCR) was used to determine the mRNA expressions of the specific transcription factors, FoxP3 and T-bet of the helper 17 cells (Th17), regulatory T cells (Treg) and T helper 1 cells (Th1). The flow cytometry was applied to determine the rates of Treg, Th1 and Th17, as well as the changes of the inflammatory reaction index, C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR). The therapeutic effects were compared between the two groups. RESULTS: After treatment, the total effective rate was 93.3% (28/30) in the observation group, which was better than 86.7% (26/30) in the control group (<0.05). After treatment, the levels of CRP, ESR, IL-6, IL-17, IL-23 and TNF-α, as well as the rate of Th17 were reduced significantly as compared with those before treatment in the observation group (all <0.05). The mRNA expressions of FoxP3 and T-bet and the rates of Treg and Th1 were increased as compared with those before treatment (all <0.05). The change degree in the observation group was significant as compared with the control group (all <0.05). In the control group, the levels of CRP, ESR, IL-6, IL-17, IL-23 and TNF-α, as well as the rate of Th17 were reduced, and the mRNA expressions of FoxP3 and T-bet and the rates of Treg and Th1 were increased after treatment. But the changes were not significant as compared with those before treatment (all >0.05). CONCLUSION The long-snake moxibustion effectively relieves the clinical symptoms in AS patients and regulates the Th17/Treg/Th1 immune imbalance. Its effect target is probably related to the modulation of the AS immune derangement and the inflammatory responses induced by immune derangement so as to achieve the dual-positive regulatory effect.
Animals ; Humans ; Lymphocyte Count ; Moxibustion ; Snakes ; Spondylitis, Ankylosing ; therapy ; T-Lymphocytes, Regulatory ; Th1 Cells ; Th17 Cells

Sensory conflict impacts postural control, yet its effect on cortico-muscular interaction remains underexplored. We aimed to investigate sensory conflict's influence on the cortico-muscular network and postural stability. We used a rotating platform and virtual reality to present subjects with congruent and incongruent sensory input, recorded EEG (electroencephalogram) and EMG (electromyogram) data, and constructed a directed connectivity network. The results suggest that, compared to sensory congruence, during sensory conflict: (1) connectivity among the sensorimotor, visual, and posterior parietal cortex generally decreases, (2) cortical control over the muscles is weakened, (3) feedback from muscles to the cortex is strengthened, and (4) the range of body sway increases and its complexity decreases. These results underline the intricate effects of sensory conflict on cortico-muscular networks. During the sensory conflict, the brain adaptively decreases the integration of conflicting information. Without this integrated information, cortical control over muscles may be lessened, whereas the muscle feedback may be enhanced in compensation.
Humans ; Muscle, Skeletal ; Electromyography/methods* ; Electroencephalography/methods* ; Brain ; Brain Mapping