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 preliminarily develop a multidimensional blood donor role scale based on role theory and systematically compare the psychosocial characteristic differences between award-winning donors and first-time donors in Guangzhou, and to provide an empirical reference for formulating differentiated donor retention strategies. Methods: A cross-sectional survey design was adopted. A random sample of award-winning donors and concurrently sampled first-time donors yielding 1 361 valid responses collected (721 from the award group, 640 from the first-time group). Exploratory factor analysis was used to assess the scale structure. Data were post-stratified and weighted according to the gender and age distributions of the general donor population. Independent samples t-tests, multivariate analysis of covariance (MANCOVA), and generalized linear models were employed to compare dimensional scores between the two groups. A paired t-test was conducted to analyze the annual donation frequency of award-winning donors before and after receiving the award. Results: Exploratory factor analysis yielded a 5-factor structure, including Role Identity and Expectations, Role Adaptation and Maintenance, Role Environment and Experience, Role Relationships and Conflict, and Role Incentives and Rewards, with a cumulative variance contribution rate of 56.43%. The scale demonstrated good internal consistency reliability (Cronbach's α=0.906). Known-group validity test showed that award-winning donors scored significantly higher than first-time donors on Role Identity and Expectations (t=4.366, P<0.001, d=0.240), Role Adaptation and Maintenance (t=5.436, P<0.001, d=0.500), and Role Relationships and Conflict (t=4.844, P<0.001, d=0.220). These differences remained significant after controlling for selected demographic variables (MANCOVA, Wilks' λ=0.943, P<0.001). Generalized linear models suggested that donation frequency was an independent predictor for these dimensions. Additionally, the annual donation frequency of award-winning donors was slightly higher after receiving the award than before (t=2.007, P=0.045). Conclusion: The preliminary blood donor role scale demonstrates acceptable reliability and validity and can effectively distinguish groups with different donation behavior characteristics. The study reveals that award-winning donors exhibit more positive psychological characteristics across multiple role identity dimensions and maintain their donation behavior after receiving an award. External incentives and internal role identity may jointly contribute to behavioral persistence. The findings provide a preliminary reference for further exploring the formation pathways of donor role identity and developing differentiated donor retention strategies.

Objective To investigate the mechanism of action of Guizhi Fuling pill in treating chro-nic prostatitis(CP)using network pharmacology and molecular docking techniques.Methods Compo-nents of Guizhi Fuling pill were collected from the Traditional Chinese Medicines Systems Pharmacolo-gy Platform(TCMSP),and target information was obtained from the SwissTarget database.Targets for chronic prostatitis were screened from the GeneCards,OMIM,CTD,and DisGeNET disease data-bases.A protein-protein interaction(PPI)network was established and analyzed.Gene ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway en-richment analysis were performed using the DAVID database.The Cytoscape software was employed to construct an association network linking the components of Guizhi Fuling Pill,their targets,and the targets of chronic prostatitis.Molecular docking was conducted using AutoDock Vina software to verify the binding stability between the components of Guizhi Fuling pill and their targets.Results After screening and deduplication in the TCMSP database,76 components of Guizhi Fuling Pill were iden-tified,and 655 component targets were retrieved from the SwissTarget database.There were 190 intersecting targets between GuizhiFuling Pill and chronic prostatitis.GO analysis indicated that Guizhi Fuling Pill may treat chronic prostatitis by participating in processes such asapoptosis,ATP binding,and signal transduction.KEGG analysis suggested that Guizhi Fuling Pill can regulate pathways such as phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)and mitogen-acti-vated protein kinase(MAPK)to intervene in chronic prostatitis.Molecular docking data demonstra-ted that the components of Guizhi Fuling pill exhibited stable conformations with their targets.Con-clusion The components of Guizhi Fuling Pill can stably bind to their targets and exert therapeutic effects on chronic prostatitis through multiple targets and pathways.

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 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 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 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.