Objective:To investigate the effects of ginkgolide B on neurological function recovery and the Wnt/β-catenin pathway after ischemic stroke in mice.Methods:Fifty-five C57/BL6 mice were selected, of which 10 mice were kept as the sham group and the remaining 45 mice were constructed as the ischemic stroke model. There were 40 mice who finally completed the modeling, and then they were randomly divided into the blank control group (GB0w), short-course administration group (GB1w), long-term administration group (GB2w), and long-term administration+antagonist group (GB2w+PRI-724), with 10 mice in each group. There was no drug intervention after MCAO in GB0w. The mice in GB1w were given ginkgolide B (10 mg/kg) 0.1 ml within 1 week after MCAO; in GB2w were given ginkgolide B (10 mg/kg) 0.1 ml within 2 weeks after MCAO; and in GB2w+PRI-724 were nasally fed ginkgolide B (10 mg/kg) 0.1 ml within 2 weeks after MCAO; and selective antagonist PRI-724 was given 3 h before administration of ginkgolide B on days 8 to 14. Neurological function scores, walking on rotor bar test scores, expression of transforming growth factor-β1 (TGF-β1), fibroblast growth factor 4 (FGF4), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), Wnt, β-catenin, and glycogen synthase kinase-3β (GSK-3β) were compared among the groups.Results:Compared with the sham group, the expressions of MDA, TNF-α, IL-6, FGF4, and GSK-3β in GB0w, GB1w, GB2w, and GB2w+ PRI-724 were increased, and the expressions of GSH-Px, SOD, TGF-β1, β-catenin, and Wnt were decreased (all P < 0.001). Compared with GB0w, the expressions of SOD, GSH-Px, TGF-β1, Wnt, and β-catenin were increased in GB1w, GB2w, and GB2w+PRI-724, and the expressions of MDA, TNF-α, IL-6, FGF4, and GSK-3β were decreased (all P < 0.001). Compared with GB1w, the expressions of GSH-Px, SOD, TGF-β 1, Wnt, and β-catenin were increased in GB2w and GB2w+PRI-724, and the expressions of IL-6, TNF-α, MDA, FGF4, and GSK-3β were decreased (all P < 0.001). Compared with GB2w, the neural function score, walking on the stick test score, and expressions of IL-6, TNF-α, FGF4, MDA, and GSK-3β were increased in GB2w+PRI-724, while the expressions of GSH-Px, TGF-β1, SOD, Wnt, and β-catenin were decreased (all P < 0.001). Conclusions:Ginkgolide B can effectively improve the neurological function of ischemic stroke mice and may be related to the Wnt/β-catenin pathway.

Objective To evaluate the predictive ability and influencing factors of individualized drug administration adjuvant decision-making system Java PK? for Desktop(JPKD)for tacrolimus blood concentration in kidney transplant recipients.Methods The monitoring data of tacrolimus blood concentration from 149 recipients early after kidney transplantation were collected.The trough blood concentration of tacrolimus was predicted by JPKD.The absolute weighted deviation and relative prediction deviation between the actual and predicted concentration were calculated.The influencing factors of the absolute weighted deviation were analyzed by univariate and multivariate logistic regression analyses,and the predictive values of these influencing factors on the accuracy of software prediction were assessed by delineating the receiver operating characteristic(ROC)curve.Results Two hundred and sixty-six samples of tacrolimus blood concentration data were collected from 149 patients.The measured blood concentration of tacrolimus was(6.5±3.0)ng/mL(1.1-16.6 ng/mL),and the predicted value calculated by JPKD was(5.6±2.5)ng/mL(1.4-14.4 ng/mL).The absolute weighted deviation of the calculated data was 28.38％,and the relative prediction deviation was-13.55％.Univariate analysis showed that gender,albumin,changes in hematocrit,cytochrome P450(CYP)3A5*3 genotype and C3435T genotype were associated with the inaccurate prediction results.Multivariate logistic regression analysis found that CYP3A5*3 genotype of AA and the changes in hematocrit were the independent risk factors affecting the accuracy of tacrolimus blood concentration predicted by JPKD.ROC curve analysis showed that when the changes in hematocrit exceeded 2.25％,the risk of inaccurate software prediction was increased.Conclusions JPKD possesses certain accuracy in predicting the blood concentration of tacrolimus in kidney transplant recipients,which may improve the qualified rate of tacrolimus blood concentration.Nevertheless,CYP3A5*3 genotype and the changes of hematocrit may affect the accuracy of predictions.

OBJECTIVE To explore the mechanism of steam-processed Polygonatum sibiricum polysaccharides(SPSP) in prophylaxis and treatment of mice with blood deficiency syndrome(BDS) by RNA sequencing(RNA-seq) technology. METHODS The mice were randomly divided into five groups(10 mice in each group), namely normal group, model group, SPSP groups(0.1, 0.4 g·kg−1), Danggui Buxue oral liquid(DOL) group. BDS model was induced in mice by acetylphenyl-hydrazine and cyclophosphamide. Blood routine, body weight and body temperature were tested after a consecutive administration for 14 d. The differential expressed genes(DEGs) related to anti-BDS by SPSP were screened through the transcriptome sequencing of the hepatic tissue in BDS mice. Functional annotation and enrichment analysis were performed to screen out the gene expression signaling pathways related to the treatment of SPSP on BDS mice. Quantitative polymerase chain reaction(qPCR) was used to verify the experiment. RESULTS Compared with the model group, SPSP(0.4 g·kg−1) could elevate the blood routine indexes such as red blood cell, white blood cell, hemoglobin, platelet, mean corpuscular hemoglobin concen-tration(P<0.01), and reverse the body weight and body temperature to normal(P<0.01 or P<0.05). The result of transcriptomic analysis showed that the underlying mechanism was mainly related to hematopoietic cell line, retinol metabolism, steroid hormone biosynthesis, platelet activation, B cell receptor signaling pathway, and leukocyte transendothelial migration, etc. The result of qPCR showed that SPSP(0.4 g·kg−1) could elevate the expression of JAK1, STAT1 and GATA1 mRNA (P<0.01 or P<0.05). CONCLUSION SPSP has therapeutic effects on BDS. The key DEGs in the treatment of BDS by SPSP are mainly related to the restoration of hematopoietic function, regulation of hormone and immune function. The mechanism of SPSP on treatment of BDS might be the regulation of JAK1/STAT1 signaling pathway.

convalescents, and to screen for broad-spectrum neutralizing antibodies against the SARS-CoV-2 RBD. Methods Using biotinylated RBD as a molecular probe, flow cytometry was employed to perform single-cell sorting of B cells from peripheral blood mononuclear cells (PBMCs) of convalescents. The obtained B cells were lysed and subjected to reverse transcription, followed by nested PCR amplification of the heavy and light chains of antibodies was conducted using random primers. The amplified products were cloned into corresponding expression vectors, and the respective matched heavy-light chain plasmids were co-transfected into 293F cells for expression. Monoclonal antibodies were then purified using Protein A column chromatography. Neutralization experiments were conducted with the wild-type (WT) pseudovirus, and antibodies with IC50<0.1 μg/mL were selected for further testing of neutralizing breadth and potency against the wild-type (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and currently prevalent pseudovirus strains (XBB, BA.5, BF.7). Results A total of 21 RBD-specific monoclonal B cells were obtained from two recovered patients, resulting in the isolation of 13 pairs of antibody light/heavy chains. Nine antibodies were successfully expressed, with P1-A1, P1-B6, and P1-B9 exhibiting IC50 values below 0.1 μg/mL against the pseudovirus of the wild-type strain (WT). Specifically, P1-B6 effectively neutralized the wild-type strain (WT), Beta variant (B.1.351), and Delta variant (B.1.617.2), with IC50 values reaching 0.01 μg/mL. P1-B9 demonstrated effective neutralization against the wild-type strain (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and Gamma variant (P.1) pseudoviruses, with IC50 values of 0.42 μg/mL, 0.63 μg/mL, 0.28 μg/mL, and 2.50 μg/mL, respectively. Additionally, P1-B6 exhibited good neutralization against BA.5 and BF.7 pseudoviruses, with IC50 values of 0.06 μg/mL and 0.09 μg/mL, respectively. Conclusions Infection with the SARS-CoV-2 WT strain can induce the generation of neutralizing antibodies with broad-spectrum activity. Generating these broadly neutralizing antibodies does not require an excessively high somatic hypermutation. The obtained antibodies can be used as candidates for SARS-CoV-2 diagnosis and prevention.

Objective:To analyze the effect of panax notoginseng saponins(PNS)on mTORC1-HIF1α signaling pathway,and to explore its effect and mechanisms on the differentiation balance of Th17/Treg cells in CD4+T cells.Methods:Isolate the spleens of C57BL/6 mice,then select CD4+T cells by magnetic beads and cultured in vitro.The optimal concentration of PNS was screened by the CCK-8,and then these cells were divided into control group and PNS treatment group(5,10 and 20 μg/ml),each gives correspond-ing drug treatment after 48 h.Afterwards,flow cytometry was used to detect differentiation of Th17/Treg cells.Real-time quantitative fluorescent PCR was used to detect the expressions of RORγt,Foxp3,mTOR,Raptor,HIF1α mRNA.ELISA was used to detect the levels of IL-17A and IL-10 in the supernatant of cell culture.Western blot was used to detect the expressions and phosphorylation levels of 4EBP1,S6K and HIF1α proteins.Results:5,10,20 μg/ml PNS could significantly inhibit Th17 cells differentiation and promote Treg cells differentiation;5,10,20 μg/ml PNS could significantly reduce the expression of RORγt mRNA,and then reduce the level of IL-17A;20 μg/ml PNS could significantly promote the expression of Foxp3 mRNA and increase the level of IL-10;10,20 μg/ml PNS could significantly decrease the phosphorylation of 4EBP1 and S6K;5,10,20 μg/ml PNS could significantly reduce the expression of HIF1α mRNA and inhibit the expression of HIF1α protein.Conclusion:Certain concentrations of PNS can inhibit the differentiation of Th17 cells in CD4+T cells,and promote the differentiation of Treg cells,which is related with modulating mTORC1-HIF1α signaling pathway.

Rapid turnover of the intestinal epithelium is a critical strategy to balance the uptake of nutrients and defend against environmental insults, whereas inappropriate death promotes the spread of inflammation. PPARα is highly expressed in the small intestine and regulates the absorption of dietary lipids. However, as a key mediator of inflammation, the impact of intestinal PPARα signaling on cell death pathways is unknown. Here, we show that Pparα deficiency of intestinal epithelium up-regulates necroptosis signals, disrupts the gut vascular barrier, and promotes LPS translocation into the liver. Intestinal Pparα deficiency drives age-related hepatic steatosis and aggravates hepatic fibrosis induced by a high-fat plus high-sucrose diet (HFHS). PPARα levels correlate with TRIM38 and MLKL in the human ileum. Inhibition of PPARα up-regulates necroptosis signals in the intestinal organoids triggered by TNF-α and LPS stimuli via TRIM38/TRIF and CREB3L3/MLKL pathways. Butyric acid ameliorates hepatic steatosis induced by intestinal Pparα deficiency through the inhibition of necroptosis. Our data suggest that intestinal PPARα is essential for the maintenance of microenvironmental homeostasis and the spread of inflammation via the gut-liver axis.

Objective:To investigate the unqualified hepatitis C virus (HCV) detection result of blood donors from the served area of blood institutions.Methods:The data related to HCV markers detected of the first and repeat blood donors were collected from the system of practice comparison for the Chinese mainland blood institutions from 2017 to 2021. The anti-HCV reactive rate and the rates of anti-HCV negative but HCV-RNA reaction and all the relationship between rates and the annual, regional and different blood donors were statistically analyzed.Results:During 2017-2021, the number of anti-HCV reactive per 100 000 blood donors decreased from 444.3 to 250.44 in the served area of 22 blood institutions ( χ2=49.677, P<0.05). The number of HCV RNA detected positive per 100 000 anti-HCV negative increased from 0.69 to 2.05 year by year, but there was no statistical significance ( χ2=0.643, P>0.05). The anti-HCV unqualified rate was significantly different among regions ( χ2=3 260.283, P<0.05). The anti-HCV unqualified rate of the first blood donors was significantly higher than that of the repeated blood donors ( F=130.993, P < 0.05). The annual number of HCV RNA detected positive per 100 000 anti-HCV negative blood samples from donors ranged from 0 to 17.28. Conclusions:The anti-HCV unqualified rate of blood donors in the served area of 22 blood institutions decreased year by year. Compared with repeated blood donors, HCV infection should be emphasized in first-time blood donors. The implementation of HCV RNA test can detect out much more HCV infections and reduce the risk of transfusion transmitted infectious HCV.

Objective:To explore the accuracy of computer optometric diopter before mydriasis in predicting artificial optometric diopter after mydriasis in the current ophthalmic standard of civil aviation recruitment, to evaluate the rationality of "0.75 D or more beyond the standard" in the current recruitment standard, and to provide a scientific basis for adjusting the recruitment diopter standard.Methods:Cluster sampling was used to select students recruited by Civil Aviation Flight University of China in Sichuan area from 2017 to 2021, including high school students and college student candidates. The values of computer optometric diopter before mydriasis and optometric diopter after mydriasis were collected, and the linear regression model was fitted to predict the optometric diopter value after mydriasis with computer optometric diopter before mydriasis as independent variable.Results:A total of 2 567 recruited students, all male, with an average age of (18.20±1.47) years were enrolled in the physical examination. There was a high correlation between computer optometric diopter before mydriasis and optometric diopter after mydriasis ( rS=0.856, P<0.001). After linear regression fitting and calculation of the predicted range of post-dilated optometric diopter, it was found that pre-dilated computer optometric diopter could explain 80.5% of the variation degree of post-dilated optometric diopter. When the computer optometric diopter before mydriasis was in the range of [-7.000,+1.500] D, the prediction accuracy of this model was ≥83.3%, and the fitting effect was good. When the computer optometric diopter before mydriasis was higher than -6.125 D, the prediction interval of 95% was higher than -4.500 D, which was beyond the recruitment standard. Conclusions:For myopic students, the predicted value of post-dilated optometric diopter exceeds the current recruitment standard if the computer optometric diopter before mydriasis exceeds -6.125 D. It is suggested to amend the corresponding item in the standard to "if the diopter exceeds 1.750 D or more, the subject can be terminated other examinations".

Objective:To explore the influence of left and right eyes and different refractive states on the variation of parameters by analyzing the difference and correlation of the diopters before and after ciliary muscle paralysis in civil flying cadets.Methods:The observational study was used. The diopters tested by computer optometry before and after ciliary palsy, including degree sphere (DS), degree cylinder (DC) and spherical equivalent (SE), were collected from 2 567 flying cadets in Sichuan area of Civil Aviation Flight University of China from 2017 to 2021. They were grouped into 5 groups by myopia degree: high myopia [-7.000 D, -6.125 D], moderate myopia [-6.000 D, -3.125 D], mild myopia [-3.000 D, -0.125 D], emmetropia [0.000 D] and mild hyperopia [+0.125 D, +3.000 D] by refractive states. The differences of DS, DC, SE in left and right eyes and different refractive states before and after ciliary muscle paralysis were compared. Paired Wilcoxon rank sum test was used for analysing the diopter difference before and after ciliary muscle paralysis, and Spearman correlation coefficient was used for analysing the correlation. Mann-whitney U test and Kruskal-Wallis H test were respectively used to determine the influence of left and right eyes and different refractive states on the changes of diopter before and after ciliary muscle paralysis. Results:After ciliary palsy, DS and SE had significant hyperopia drift compared with those before ciliary palsy with statistically significant differences ( Z=-35.95, -36.05, both P<0.001). The changes of DS, DC and SE were not affected by eye gender (all P>0.05). The changes of DS and SE were affected by the refractive states ( H c=217.33, 237.59, both P<0.001); The change of DC was not affected by the refractive states ( H c=8.22, P=0.084). The changes of DS and SE before and after ciliary palsy were mainly negative, within the range of (-0.525~0.075) D. The higher the degrees of myopia, the more obvious SE hyperopia drift. The correlation of SE values before and after cycloplegia was high ( r s=0.870-0.877). The SE correlation was the weakest in the people with high myopia ( r s=0.288), but the strongest for mild myopia ( r s=0.767). Conclusions:The computerized optometry can be used in large-scale initial screening examination of recruitment physical examination and the measured column diopter value can be used as a reference.

Objective:To explore the accuracy of computer optometric diopter before mydriasis in predicting artificial optometric diopter after mydriasis in the current ophthalmic standard of civil aviation recruitment, to evaluate the rationality of "0.75 D or more beyond the standard" in the current recruitment standard, and to provide a scientific basis for adjusting the recruitment diopter standard.Methods:Cluster sampling was used to select students recruited by Civil Aviation Flight University of China in Sichuan area from 2017 to 2021, including high school students and college student candidates. The values of computer optometric diopter before mydriasis and optometric diopter after mydriasis were collected, and the linear regression model was fitted to predict the optometric diopter value after mydriasis with computer optometric diopter before mydriasis as independent variable.Results:A total of 2 567 recruited students, all male, with an average age of (18.20±1.47) years were enrolled in the physical examination. There was a high correlation between computer optometric diopter before mydriasis and optometric diopter after mydriasis ( rS=0.856, P<0.001). After linear regression fitting and calculation of the predicted range of post-dilated optometric diopter, it was found that pre-dilated computer optometric diopter could explain 80.5% of the variation degree of post-dilated optometric diopter. When the computer optometric diopter before mydriasis was in the range of [-7.000,+1.500] D, the prediction accuracy of this model was ≥83.3%, and the fitting effect was good. When the computer optometric diopter before mydriasis was higher than -6.125 D, the prediction interval of 95% was higher than -4.500 D, which was beyond the recruitment standard. Conclusions:For myopic students, the predicted value of post-dilated optometric diopter exceeds the current recruitment standard if the computer optometric diopter before mydriasis exceeds -6.125 D. It is suggested to amend the corresponding item in the standard to "if the diopter exceeds 1.750 D or more, the subject can be terminated other examinations".