ObjectiveTo observe the effect of Yifei Jianpi prescription on the of signal transducer and activator of transcription protein 1 （STAT1）/interferon regulatory factor 3 （IRF3） signaling pathway in a pneumonia model induced by lipopolysaccharide （LPS） and to explore the mechanism of Yifei Jianpi prescription in improving lung immune and inflammatory responses. MethodsSixty male SPF SD rats were used in this study. Ten rats were randomly assigned to the normal control group， and the remaining 50 were instilled with LPS in the trachea to establish a pneumonia model. After successful modeling， the rats were randomly divided into the model group， dexamethasone group （0.5 mg·kg^-1）， and Yifei Jianpi prescription high-dose （12 mg·kg^-1）， medium-dose （6 mg·kg^-1）， and low-dose （3 mg·kg^-1） groups， with 10 rats in each group. Treatment was administered once daily， and the normal control and model groups received the same volume of normal saline. After 14 days， flow cytometry was used to detect the classification of whole blood lymphocytes. Enzyme-linked immunosorbent assay （ELISA） was used to measure serum levels of immunoglobulin G （IgG）， immunoglobulin A （IgA）， immunoglobulin M （IgM）， and the content of tumor necrosis factor-α （TNF-α）， interleukin-8 （IL-8）， interleukin-6 （IL-6）， and interleukin-10 （IL-10） in alveolar lavage fluid （BALF）. Hematoxylin-eosin （HE） staining was used to observe lung tissue pathology and score the damage. Thymus weight， spleen weight， and wet-to-dry weight ratio （W/D） were recorded. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of STAT1， IRF3， IL-6， and interferon-alpha （IFN-α） in lung tissues， while Western blot was performed to assess the protein expression of STAT1， IRF3， IL-6， and IFN-α. ResultsCompared with the normal control group， the model group showed significantly increased proportion of B lymphocytes in peripheral blood， decreased proportions of NK cells and CD4⁺/CD8⁺ （P<0.05， P<0.01）， decreased serum levels of IgG and IgA， significantly increased IgM levels （P<0.01）， significantly elevated content of TNF-α， IL-6， and IL-8 in BALF， and significantly decreased IL-10 levels （P<0.01）. Lung tissue damage was evident， with significant increases in thymus and spleen weights and a higher W/D ratio （P<0.01）. The mRNA and protein expression of STAT1， IRF3， IFN-α， and IL-6 in lung tissues was significantly upregulated （P<0.05，P<0.01）. Compared with the model group， the Yifei Jianpi prescription groups showed significantly reduced proportions of B lymphocytes in peripheral blood， increased proportions of NK cells and CD4⁺/CD8⁺ ratios （P<0.05， P<0.01）， significantly increased serum levels of IgG and IgA， significantly decreased IgM levels （P<0.05， P<0.01）， significantly reduced levels of TNF-α， IL-6， and IL-8 in BALF， and significantly increased IL-10 levels （P<0.01）. Lung tissue damage was alleviated， thymus and spleen weights were significantly reduced， and the W/D ratio was markedly decreased （P<0.01）. The mRNA and protein expression of STAT1， IRF3， IFN-α， and IL-6 in lung tissues was significantly downregulated （P<0.05， P<0.01）. ConclusionYifei Jianpi prescription can alleviate lung tissue damage and improve immune and inflammatory responses in LPS-induced pneumonia rats. The mechanism may be related to the inhibition of STAT1/IRF3 signaling pathway activation.

Objective To analyze the causes of HIV-2 specific bands in the Western blot (WB) tests and to understand previous HIV-2 infection status in this city. Methods A total of 68 samples with HIV-2 specific bands in WB were analyzed using two confirmatory reagents. The test results were further analyzed in combination with epidemiological data, nucleic acid testing and gene sequencing. Results When tested with MP reagent, 66 samples (97.06%) were found to be positive for HIV-2 antibody, while the other two were negative or undetermined for HIV-2 antibody. When tested with MIKROGEN reagent, 67 samples (98.53%) were found to be positive for HIV-1 antibody, and one sample was negative for HIV-1 antibody. Further HIV-1 nucleic acid testing was conducted on these samples, and all 68 samples tested positive for HIV-1 RNA, with the results all exceeding 5,000 copies/ml. After BLAST comparison, it was found that the homology similarity of 68 samples to the HIV-1 reference strain sequence was >90%, but there was no similarity with the HIV-2 reference strain sequence. Conclusion The results of the serological test, nucleic acid test and gene sequencing of the 68 samples all have indicated HIV-1 infection. Combined with the epidemiological data, it can be concluded that the double reaction of HIV-1 and HIV-2 antibodies in WB tests of these 68 samples is very likely to be a non-specific cross-reaction rather than HIV-2 infection. This study indicates that no HIV-2 infection cases have been found in Chengdu so far.

Objective:To investigate the effects of an immersive virtual reality system for nasal anatomy in improving the learning interest and theoretical level of eight-year program medical students for nasal anatomy in otolaryngology.Methods:A total of 57 medical students of the 2019 eight-year program in Peking University Health Science Center were divided into three groups naturally, with 19 students in each group. Two of the groups (38 students) were randomly selected as experimental group to receive traditional teaching in combination with the immersive virtual reality system for nasal anatomy, while the remaining group (19 students) served as control group to receive traditional teaching. The teaching effectiveness of the two groups was evaluated through questionnaire surveys and theoretical assessment scores in otolaryngology and nasal anatomy. SPSS 27.0 was used to perform statistical analysis. The two groups were compared using the independent samples t-test and the chi-squared test. Results:According to the survey results, both groups considered rhinology theory and anatomy learning as a challenge during rhinology internship. A significantly higher proportion of students in the experimental group believed that they gained the greatest achievement in rhinology theory and anatomy learning during internship, as compared with the control group (33/38 vs. 12/19, P=0.039). The theoretical score for nasal anatomy-related assessment was significantly higher in the experimental group than in the control group [(19.0±1.3) vs. (18.1±1.3), P=0.024]. Conclusions:The use of the immersive virtual reality system for nasal anatomy in otolaryngology teaching can promote eight-year program medical students' interest in learning and improve teaching effectiveness.

Objective:To observe any effect of transcranial direct current stimulation (tDCS) on the cognition of stroke survivors and the integrity of their white matter fibers.Methods:Thirty persons with post-stroke cognitive impairment (PSCI) were randomly divided into an experimental group ( n=15) and a control group ( n=15). In addition to basic drug therapy and routine cognition training, the experimental group received 20 minutes of tDCS daily, 5 days per week for 3 weeks, while the control group received sham tDCS stimulation. Before and after the treatment, both groups′ cognitive functioning was evaluated using the mini-mental state examination (MMSE) and the Montreal cognitive assessment scale (MoCA). Their ability in the activities of daily living (ADL) was quantified using the modified Barthel index (MBI). Diffusion tensor imaging (DTI) was employed to observe any changes in the integrity of their white matter fibers. Results:The average MMSE, MOCA and MBI scores of both groups had improved significantly after the treatment, but the improvement in the experimental group was significantly greater than among the controls. The average fractional anisotroposy value of the affected inferior fronto-occipital fasciculus in both groups was positively correlated with the group′s average MMSE score and MoCA score.Conclusion:tDCS can effectively improve the cognition and functioning in the activities of daily living of stroke survivors. Its mechanism may be related to improving the integrity of the white matter fibers involved.

Objective:To evaluate the effect of propofol on neuronal activity in medial prefrontal cortex (mPFC) during social behavior in sleep-deprived rats.Methods:Sixty SPF healthy male Sprague-Dawley rats, aged 8 weeks, weighing 200-250 g, were divided into 3 groups ( n=20 each) using a random number table method: control group (group Con), chronic sleep deprivation plus natural sleep group (group CSD+ NS), and chronic sleep deprivation plus propofol group (CSD+ Pro). Sleep deprivation model was developed by the modified multiple platform method, and the rats were placed in the sleep-deprivation tank for 20 h a day (14: 00-10: 00) and allowed to sleep naturally for 4 h (10: 00-14: 00) a day for 28 consecutive days.Propofol 40 mg/kg was intraperitoneally injected after sleep deprivation for 28 consecutive days in group CSD+ Pro, while the equal volume of 10% fat emulsion was given instead in group C and group CSD+ NS.Electroencephalographic recordings in cerebral cortical regions were performed on the days 1st, 14th and 28th after sleep deprivation.The apoptotic neurons in mPFC were detected using TUNEL method after the end of sleep deprivation, and the apoptosis index was calculated.A three chamber sociability test was used to detect the social behavior of rats, and local field potential signals in mPFC were collected. Results:Compared with group Con, the percentage of rapid eye movement sleep was significantly increased, the sniffing time preference coefficients in the 2 stages were reduced, the percentage of the β waves and θ waves-band power in mPFC during the social sniffing process was decreased, and the apoptosis index of neurons in mPFC was increased in group CSD+ NS ( P<0.05). Compared with group CSD+ NS, the percentage of rapid eye movement sleep was significantly increased, the sniffing time preference coefficient in the 2 stages was increased, and the percentage of β waves and θ waves-band power in mPFC during the social sniffing process was increased, and the apoptosis index of neurons in mPFC was decreased in group CSD+ Pro ( P<0.05). Conclusions:Propofol inhibits the apoptosis in neurons in mPFC and increases β and θ waves in the mPFC during social interaction after sleep deprivation in sleep-deprived rats, which is helpful in improving sleep deprivation-induced social disorder.

Objective:To evaluate the effect of botulinum neurotoxin A (BoNT/A) on prevention of chronic migraine (CM) in mice and explore the potential mechanism.Methods:Twenty-four male C57BL/6 mice were randomly divided into control group, nitroglycerin (NTG) group, and BoNT/A+NTG group ( n=8). Mice in the latter two groups were intraperitoneally injected with 10 mg/kg NTG on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments to establish CM models. Mice in the BoNT/A+NTG group were injected with 0.18 U/100 μL BoNT/A one h before the first injection of NTG. Mice in the control group were injected with the same dose of normal saline. Basal mechanical withdrawal threshold (MWT) and evoked MWT 2 h after NTG in the facial and hindpaw regions on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments were evaluated by von Frey filament test. The motor function of mice 2 h after NTG injection was tested by rotarod test on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments. On 9 th d of experiments, the mice were sacrified; the calcitonin gene-related peptide (CGRP), synaptosomal-associated protein 25 (SNAP25), glial fibrillary acidic protein (GFAP) and TRP channel protein expressions in the trigeminal ganglia (TG) and trigeminal nucleus caudalis (TNC), and NOD-like receptor protein 3 (NLRP3) inflammatory factor pathway-related protein expressions in TNC were detected by Western blotting; real-time quantitative PCR (RT-qPCR) was used to detect the NLRP3 inflammatory factor pathway-related mRNA expressions in TNC. The CGRP expression in TNC was detected by immunofluorescent staining. Results:(1) As compared with the control group, the NTG group had significantly decreased basal facial MWT on the 7 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased basal facial MWT on the 7 th and 9 th d of experiments ( P<0.05). As compared with the control group, the NTG group had significantly decreased evoked facial MWT on the 5 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased evoked facial MWT on the 5 th and 9 th d of experiments ( P<0.05). As compared with the control group, the NTG group had significantly decreased basal and evoked MWT in the hindpaw regions on the 3 rd, 5 th, 7 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased basal and evoked MWT in the hindpaw regions on the 3 rd, 5 th, 7 th and 9 th d of experiments ( P<0.05). (2) There was no significant difference in running time on rotarod among the three groups ( P>0.05). (3)Western blotting results showed that as compared with those in the control group, the CGRP and SNAP25 protein expressions were significantly increased in TG of the NTG group ( P<0.05); and those in the BoNT/A+NTG group were significantly decreased as compared with those in the NTG group ( P<0.05). As compared with those in the control group, the CGRP and NLRP3 protein expressions were significantly increased in TNC of NTG group ( P<0.05); and those in the BoNT/A+NTG group were significantly decreased as compared with those in the NTG group ( P<0.05). (4)RT-qPCR results showed that as compared with that in the control group, the IL-1β mRNA expression in TNC of the NTG group was significantly increased ( P<0.05), and that in the BoNT/A prevention group was statistically decreased as compared with that in the NTG group ( P<0.05). (5) Immunofluorescent staining results showed that as compared with that in the control group, the CGRP expression in TNC of the NTG group was significantly increased, and that in the BoNT/A+NTG group was significantly decreased as compared with that in the NTG group ( P<0.05). Conclusion:BoNT/A can reduce the SNAP25 expression in TG, reduce the CGRP release in TG and TNC, and prevent CM onset; BoNT/A can regulate NLRP3 level in TNC.

Objective:To evaluate the changes in glucose metabolism in the prefrontal cortex during long-term cognitive dysfunction induced by neuropathic pain in developing rats.Methods:SPF healthy male Sprague-Dawley rats, aged 4 weeks, weighing 80-100 g, were used in this study.The model of neuropathic pain was established by using spared nerve injury in anesthetized rats.The mechanical paw withdrawal threshold (MWT) was measured at 1 day before establishing the model (T 0) and 1, 3, 7, 14, 28, 42 and 56 days after establishing the model (T 1-7). According to the results of MWT compared between T 5 and T 0, the rats were divided into neuropathic pain group (group NP) and non-neuropathic pain group (group NNP). Open field test and novel object recognition test were performed at T 7 to assess anxiety-like behavior and cognitive function.Positron emission tomography/computed tomography imaging was performed to determine the standard uptake value of 18F-fluorodeoxyglucose in the prefrontal cortex.Then the rats were sacrificed, and prefrontal cortex was removed for determination of the expression of glucose transporter 3 using Western blot and immunofluorescence. Results:Compared with the baseline at T 0, the MWT at T 1-2 in group NNP and at T 1-7 in group NP were significantly decreased ( P<0.05). Compared with group NNP, the MWT at T 1-7 were significantly decreased, the time of staying at the central region at T 7 was shortened, the percentage of time for exploring the novel object was decreased, the percentage of novel object exploration was decreased, the standard uptake value of 18F-fluorodeoxyglucose in prefrontal cortex was decreased, and the expression of glucose transporter 3 in prefrontal cortex was down-regulated in group NP ( P<0.05). Conclusion:Long-term cognitive dysfunction induced by neuropathic pain may be related to decreased glucose metabolism in the prefrontal cortex of the developing rats.

Targeted programmed death-ligand 1 (PD-L1) and CXC chemokine receptor type 4 (CXCR4), gene sequences encoding anti-PD-L1 nanobody and anti-CXCR4 nanobody were cloned into the pET-22b (+) vector to construct recombinant expression plasmid of anti-PD-L1&CXCR4 bispecific nanobody, which was connected with 6 × His tag and transformed into E.coli BL21 (DE3). The expressed proteins were then found to exist as a soluble form in the supernatant of bacterial lysate after induction of IPTG.Three purification methods were used to obtain the target protein in order to improve the yield and purity of the bispecific nanobody.The bacterial supernatant was separated and purified by His Trap FF affinity chromatographic column.The target protein output could exceed 1 mg/L, and the product purity could reach up to 97%.Besides, the anti-PD-L1&CXCR4 bispecific nanobody shows a specific binding ability to two antigens on the cell surface, enhancing the cytotoxicity of IL-2 activated human peripheral blood mononuclear cells (PBMC) to tumor cell line AsPC-1, which lays the foundation for further evaluation of its drug efficacy in vivo.

Objective:To explore the effect of exercise program based on somatosensory interactive games in elderly patients with stable chronic obstructive pulmonary disease (COPD) .Methods:From June 2017 to October 2019, convenience sampling was used to select 130 elderly patients with stable COPD who were admitted to West China Hospital of Sichuan University. Patients were randomly divided into intervention group and control group, 65 cases in each group. Control group carried traditional exercise training, and intervention group implemented somatosensory game interactive training. We compared the exercise compliance, physical activity and lung function of the two groups of patients. In the end, 61 patients in intervention group and 58 patients in control group completed the study. SPSS 25.0 statistical software was used for statistical analysis.Results:After 18 weeks of intervention, the exercise compliance rate of patients in intervention group was 83.61% (51/61) , which was higher than 67.24% (39/58) in control group, the difference was statistically significant (χ 2=4.320, P<0.05) . After 18 weeks of intervention, the 6-minute walking test distance of intervention group was (523.27±29.12) m, and the blood oxygen saturation was (97.00±1.35) %, and the forced expiratory volume in the first second divided by forced vital capacity was (0.90±0.15) , higher than those of control group, the differences were statistically significant ( t=-4.770, -2.865, -3.142; P<0.05) . Conclusions:Somatosensory game interactive training can improve the exercise compliance, physical activity and lung function of elderly patients with COPD in stable period, which has promotion value.

Objective:To evaluate the relationship between P2X 7 receptors and nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3)/interleukin-1beta (IL-1β) pathway in spinal neurons in the development of inflammatory pain (IP) in rats. Methods:SPF healthy adult male Wistar rats, weighing 180-220 g, were used in this study.Forty rats in which intrathecal catheters were successfully implanted were divided into 5 groups ( n=8 each) using a random number table method: control group (group CON), group IP, IP plus dimethyl sulfoxide (DMSO) group (group IP-DMSO), IP plus P2X 7 receptor antagonist A740003 group (group IP-A) and IP plus P2X 7 receptor agonist ATP group (group IP-ATP). Rats were anesthetized with pentobarbital sodium 40 mg/kg.IP was induced by injecting complete Freund′s adjuvant 50 μl into the right ankle joint cavity, while group CON was injected with the equal volume of normal saline instead.On 1 day before establishing the model, immediately after establishing the model, and on 1, 2 and 3 days after establishing the model, 1% DMSO 10 μl was intrathecally injected once a day in group IP-DMSO, A740003 0.1 nmol(dissolved in DMSO 10 μl) was intrathecally injected once a day in group IP-A, and ATP 150 nmol(dissolved in DMSO 10 μl) was injected intrathecally once a day in group IP-ATP.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured on 3 days after establishing the model.Enzyme-linked immunosorbent assay was used to determine the prostaglandin E2 (PGE2) concentrations in right ankle tissues and IL-1β concentrations in cerebrospinal fluid (CSF). Then rats were sacrificed, and the lumber segments (L 4-6) of the spinal cord were removed for determination of the expression of NLRP3, casepase-1, IL-1β (by Western blot) and co-expression of P2X 7 receptors with neuron-specific nucleoprotein (NeuN) and NLRP3 and with NeuN (by immunofluorescence). Results:Compared with group CON, PGE2 contents in ankle tissues were significantly increased in group IP, and the MWT was significantly decreased, the TWL was shortened, the concentrations of IL-1β in CSF were increased, and the expression of NLRP3, caspase-1 and IL-1β was up-regulated in the other four groups ( P<0.05). Compared with group IP, the MWT was significantly increased, the TWL was prolonged, the concentrations of IL-1β in CSF were decreased, and the expression of NLRP3, caspase-1 and IL-1β was down-regulated in group IP-A ( P<0.05), the MWT was significantly decreased, TWL was shortened, the concentrations of IL-1β in CSF were increased, and the expression of NLRP3, caspase-1 and IL-1β was up-regulated in group IP-ATP ( P<0.05), and no significant change was found in the parameters mentioned above in group IP-DMSO ( P>0.05). P2X 7 was co-expressed with NeuN, and NLRP3 was co-expressed with NeuN. Conclusion:P2X 7 receptors in spinal neurons are involved in the development of inflammatory pain by activating NLRP3/IL-1β signaling pathway in rats.