Objective To examine the serological and molecular prevalence as well as genotype characteristics of human Parvovirus B19 blood donors in Lanzhou,and to provide evidence for developing a screening strategy to reduce the risk of blood transfusion transmission.Methods A total of 5 722 blood samples collected from Lanzhou blood donors from April 2023 to October 2023 were tested for B19 DNA using real-time quantitative PCR(qRT-PCR).Additionally,383 samples were screened for anti-B19 IgG and anti-B19 IgM using synchronous enzyme-linked immunoassay(ELISA).Viral load and VP1 sequencing were conducted on the B19 DNA-positive samples and the Neighbor-Joining(N-J)method was used to construct an evolutionary tree for the sequenced samples.Results The prevalence of human Parvovirus B19 DNA,IgG antibody and IgM antibody was 0.47%(27/5 722),25.59%(98/383)and 0.26%(1/383),respectively,and the samples positive for B19 DNA,IgG antibody and IgM antibody were 0.26%(1/383).The co-positivity rate for B19 DNA and IgG antibody was 6.27%(24/383),while the positivity rates for B19 DNA or IgG antibody alone were 0.52%(2/383)and 19.06%(73/383),respectively.Viral loads ranged from 4.24 IU/ml to 5.67×102 IU/ml,all below 104 IU/ml.There was no statistical significance in the positive rate of B19 DNA in gender(χ2=0.86,P=0.35),but there was statistical significance in the positive rate of B19 DNA among all age groups(χ2=8.00,P=0.02).The highest positive rate of B19 DNA was 0.65%in the 18～30 age group.There was statistical significance in the positive rate of B19 IgG antibody in gender(χ2=5.03,P=0.02),but there was no statistical significance in the positive rate of B19 IgG antibody among all age groups(χ2=0.51,P=0.77).The highest positive rate of B19 IgG antibody was 29.09%in the age group of 41 to 60.There was no significant difference in the positive rate of B19 IgM antibody in gender(χ2=2.84,P=0.09).The highest positive rate of B19 IgM was 3.85%in the age group of 18～30 years old.Based on the VP1 sequence,the phylogenetic tree revealed that B19 strains in Lanzhou formed a distinct cladistic lineage within genotype 1,predominantly represented by genotype 1b.Conclusion The prevalence of B19 DNA and IgM antibodies among blood donors in the Lanzhou area is low,and so is the viral load.Therefore the risk of transmitting B19 through blood transfusion is relatively small.Since the prevalence of B19 IgG antibody is high,it is suggested to closely monitor the transmission situation in the area,regularly monitor the prevalence of B19 among blood donors,and track the situation of B19 DNA-positive blood donors to recipients to ensure the safety of clinical blood transfusion.

BACKGROUND:Cognitive impairment is the most common complication after stroke,and its severity is closely related to the patient's prognosis.The prognosis of patients can be significantly improved if the severity of their cognitive impairment is recognized and targeted early.OBJECTIVE:To initially explore potential biomarkers affecting the progression of post-stroke cognitive impairment,thereby providing a richer and unique reference for the study of their pathophysiological mechanisms.METHODS:Using ultra performance liquid chromatography-mass spectrometry,non-targeted metabolomics analysis was conducted on serum samples from patients with mild and moderate post-stroke cognitive impairment to identify differential metabolites between the two groups.To further validate the diagnostic efficacy of the differential metabolites,the receiver operating characteristic curve analysis was used to evaluate their accuracy and sensitivity in distinguishing disease severity.In addition,pathway analysis was conducted on the differential metabolites.RESULTS AND CONCLUSION:(1)There were significant differences in metabolic profiles between patients with mild and moderate post-stroke cognitive impairment,and 9 differential metabolites were screened by the receiver operating characteristic curve.(2)Differential metabolite pathway analysis revealed that the metabolic pathways affecting disease progression in patients with mild-to-moderate post-stroke cognitive impairment included tryptophan metabolism,D-amino acid metabolism,biotin metabolism,retinol metabolism,aminoacyl-tRNA biosynthesis,lysine degradation,protein digestion and uptake,pyrimidine metabolism,cysteine and methionine metabolism,ABC transporter proteins,amino acid biosynthesis,and 2-oxocarboxylic acid metabolism.To conclude,9 potential biomarkers affecting disease progression in patients with mild-to-moderate post-stroke cognitive impairment have been identified,involving 12 metabolic pathways including tryptophan metabolism,D-amino acid metabolism and retinol metabolism.

Objective:To investigate the role of inhibitor of differentiation 2(Id2)in inducing the generation of central memory T(Tcm)cells and enhancing the anti-tumor persistence of T cells.Methods:CD8+na?ve T cells were sorted with magnetic beads and then co-cultured with carcinoembryonic antigen(CEA)-loaded dendritic cells(DCs).These cells were induced into effector T(Teff)or Tcm cells by interleukin-2(IL-2)or IL-7/15/21/23,respectively.The mRNA and protein expression of Id2 and Id3 in T cells were detected using qPCR and WB,respectively.Id2 gene in T cells was knocked down using lentivirus,and the T cell memory phenotype was analyzed by flow cytometry.The expression of PI3K/AKT pathway-related proteins was examined by WB.The extracellular acidification rate(ECAR)and oxygen consumption rate(OCR)were assessed using a Seahorse extracellular flux analyzer.A zebrafish colorectal cancer HCT116 xenograft model was employed to analyze the anti-tumor differences between Teff and Tcm cells.The effect of Id2 gene knockdown in Tcm cells(Tcm-shId2)on the growth inhibition of secondary xenografts was also observed.Results:Tcm cells exhibited high expression of Id3 mRNA(P<0.05),whereas Teff cells showed high expression of Id2 mRNA(P<0.001).Tcm cells with Id2 knockdown(Tcm-shId2)were successfully constructed,showing significantly upregulated Id3 expression.Knockdown of Id2 promoted the formation of Tcm cell(P<0.05).Tcm-shId2 cells underwent metabolic reprogramming via the PI3K/AKT pathway,which effectively suppressed the growth of colorectal cancer xenografts in zebrafish and also produced significant inhibitory effects on secondary tumor growth(P<0.01).Conclusion:Id2 gene may regulate T cell metabolism through the PI3K/AKT signaling pathway,promoting the differentiation of CD8+T cells into Tcm cells and effectively inhibiting the growth of colorectal cancer xenografts.

Objective To examine the serological and molecular prevalence as well as genotype characteristics of human Parvovirus B19 blood donors in Lanzhou,and to provide evidence for developing a screening strategy to reduce the risk of blood transfusion transmission.Methods A total of 5 722 blood samples collected from Lanzhou blood donors from April 2023 to October 2023 were tested for B19 DNA using real-time quantitative PCR(qRT-PCR).Additionally,383 samples were screened for anti-B19 IgG and anti-B19 IgM using synchronous enzyme-linked immunoassay(ELISA).Viral load and VP1 sequencing were conducted on the B19 DNA-positive samples and the Neighbor-Joining(N-J)method was used to construct an evolutionary tree for the sequenced samples.Results The prevalence of human Parvovirus B19 DNA,IgG antibody and IgM antibody was 0.47%(27/5 722),25.59%(98/383)and 0.26%(1/383),respectively,and the samples positive for B19 DNA,IgG antibody and IgM antibody were 0.26%(1/383).The co-positivity rate for B19 DNA and IgG antibody was 6.27%(24/383),while the positivity rates for B19 DNA or IgG antibody alone were 0.52%(2/383)and 19.06%(73/383),respectively.Viral loads ranged from 4.24 IU/ml to 5.67×102 IU/ml,all below 104 IU/ml.There was no statistical significance in the positive rate of B19 DNA in gender(χ2=0.86,P=0.35),but there was statistical significance in the positive rate of B19 DNA among all age groups(χ2=8.00,P=0.02).The highest positive rate of B19 DNA was 0.65%in the 18～30 age group.There was statistical significance in the positive rate of B19 IgG antibody in gender(χ2=5.03,P=0.02),but there was no statistical significance in the positive rate of B19 IgG antibody among all age groups(χ2=0.51,P=0.77).The highest positive rate of B19 IgG antibody was 29.09%in the age group of 41 to 60.There was no significant difference in the positive rate of B19 IgM antibody in gender(χ2=2.84,P=0.09).The highest positive rate of B19 IgM was 3.85%in the age group of 18～30 years old.Based on the VP1 sequence,the phylogenetic tree revealed that B19 strains in Lanzhou formed a distinct cladistic lineage within genotype 1,predominantly represented by genotype 1b.Conclusion The prevalence of B19 DNA and IgM antibodies among blood donors in the Lanzhou area is low,and so is the viral load.Therefore the risk of transmitting B19 through blood transfusion is relatively small.Since the prevalence of B19 IgG antibody is high,it is suggested to closely monitor the transmission situation in the area,regularly monitor the prevalence of B19 among blood donors,and track the situation of B19 DNA-positive blood donors to recipients to ensure the safety of clinical blood transfusion.

BACKGROUND:Cognitive impairment is the most common complication after stroke,and its severity is closely related to the patient's prognosis.The prognosis of patients can be significantly improved if the severity of their cognitive impairment is recognized and targeted early.OBJECTIVE:To initially explore potential biomarkers affecting the progression of post-stroke cognitive impairment,thereby providing a richer and unique reference for the study of their pathophysiological mechanisms.METHODS:Using ultra performance liquid chromatography-mass spectrometry,non-targeted metabolomics analysis was conducted on serum samples from patients with mild and moderate post-stroke cognitive impairment to identify differential metabolites between the two groups.To further validate the diagnostic efficacy of the differential metabolites,the receiver operating characteristic curve analysis was used to evaluate their accuracy and sensitivity in distinguishing disease severity.In addition,pathway analysis was conducted on the differential metabolites.RESULTS AND CONCLUSION:(1)There were significant differences in metabolic profiles between patients with mild and moderate post-stroke cognitive impairment,and 9 differential metabolites were screened by the receiver operating characteristic curve.(2)Differential metabolite pathway analysis revealed that the metabolic pathways affecting disease progression in patients with mild-to-moderate post-stroke cognitive impairment included tryptophan metabolism,D-amino acid metabolism,biotin metabolism,retinol metabolism,aminoacyl-tRNA biosynthesis,lysine degradation,protein digestion and uptake,pyrimidine metabolism,cysteine and methionine metabolism,ABC transporter proteins,amino acid biosynthesis,and 2-oxocarboxylic acid metabolism.To conclude,9 potential biomarkers affecting disease progression in patients with mild-to-moderate post-stroke cognitive impairment have been identified,involving 12 metabolic pathways including tryptophan metabolism,D-amino acid metabolism and retinol metabolism.

ObjectiveTo establish a qualitative and quantitative analysis method for chemical constituents in Liu Junzitang（LJZT）， and to clarify its material basis. MethodThe chemical constituents in LJZT were analyzed by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， and the resulting compounds were identified by using databases， such as MassBank， PubChem， ChemSpider， Traditional Chinese Medicine Systems Pharmacology Database and Analytical Platform（TCMSP）， and by combining with relevant literature. UPLC was used to establish a quantitative method for analysis of 9 compounds in LJZT， including liquiritin， hesperidin， lobetyolin， liquiritigenin， glycyrrhizic acid， nobiletin， tangeretin， atractylenolide Ⅱ and Ⅰ. ResultBy combining the relevant literature， database and MS information， a total of 79 compounds were identified from LJZT， including 31 flavonoids， 15 terpenoids， 14 nitrogen-containing compounds， 6 phenylpropanoids， 6 organic acids and 7 other compounds. The established quantitative analytical method for the nine representative components showed good linearity within their respective linear ranges， and the precision， stability， reproducibility and recovery were in accordance with the requirements. The quantitative results showed that the contents of liquiritin， hesperidin， lobetyolin， liquiritigenin， glycyrrhizic acid， nobiletin， tangeretin， atractylenolide Ⅱ and Ⅰ in LJZT were 0.376 5， 2.602 1， 0.082 6， 0.128 1， 1.778 6， 0.015 7， 0.006 7， 0.030 4， 0.003 2 mg·g^-1， respectively. ConclusionThe established method can quickly， sensitively and accurately analyze the chemical constituents in LJZT， clarify that the material basis of LJZT is mainly flavonoids， terpenoids and nitrogen-containing compounds， and simultaneously determine the contents of the 9 components， which can lay a foundation for the research on quality control， mechanism and clinical application of LJZT.

Objective To evaluate the diagnostic value of myocardial work related parameters in coronary ischemia patients with coronary artery disease(CAD)coronary ischemia using non-invasive left ventricular pressure strain loop(PSL),taking fraction flow reservation(FFR)as the gold standard.Methods From December 2020 to December 2021,53 clinically suspected CAD patients were prospectively enrolled.All patients underwent echocardiography,invasive coronary angiography and FFR measurement.According to the results of coronary angiography,patients were divided into myocardial ischemia group(n=24,FFR≤0.80)and non-myocardial ischemia group(n=29,FFR＞0.80).PSL was used for off-line analysis to obtain the global work index(GWI),global constructive work(GCW),global wasted work(GWW),global work efficiency(GWE),global positive work(GPW),and global systolic constructive work(GSCW)and other myocardial work parameters.The differences of parameter values between the two groups were compared.The diagnostic efficacy of work parameters in myocardial ischemia was analyzed by ROC curve.Results Compared with the non-myocardial ischemia group,GWI,GCW,GPW and GSCW were significantly decreased in the myocardial ischemia group at the 18-,16-,and 12-segment levels(P＜0.001).The ROC curve showed that the AUC results of GWI,GCW,GPW,GSCW at the 18-segment level were 0.803(95％CI 0.679-0.927),0.807(95％CI 0.687-0.928),0.822(95％CI 0.708-0.936),0.819(95％CI 0.703-0.935).The optimal cut-off value of GWI was 1 676.3 mmHg％,and the sensitivity,specificity and accuracy of predicting myocardial ischemia were 70.8％,86.2％and 79.2％,respectively.The optimal cut-off value of GCW was 1 999.4 mmHg％,and the sensitivity,specificity and accuracy of predicting myocardial ischemia were 75.0％,82.8％and 79.2％,respectively.Conclusions Analyzing myocardial work using PSL has good significance for screening suspected myocardial ischemia in CAD patients.

Objective:To explore the pathogenesis of flunarizine-induced parkinsonism from gut-brain axis perspective.Methods:Thirty male C57BL/6 mice were randomly divided into control group and flunarizine group ( n=15). Mice in the control group were given 0.1 mL 50% polyethylene glycol 400+50% saline by gavage once/d for 2 weeks, while mice in the flunarizine group were given 6 mg/mL flunarizine+50% polyethylene glycol 400+50% saline by gavage at a daily dose of 30 mg/kg for 2 weeks. Body mass was recorded 1, 3, 5, 7, 10 and 14 d after drug administration, and motor function was assessed by rotarod test 14 d after drug administration; 16s RNA sequencing was performed in the feces to observe the intestinal flora; intestinal transit function was detected by Evans blue by gavage; and then, the mice were sacrificed and homogenate or frozen sections (brain and intestinal tissues) were prepared; dopamine-ergic neuron expression was detected by Western blotting; RT-qPCR was applied to detect the expressions of inflammatory factors in the substantia nigra, and immunofluorescent staining was used to detect the expressions of ZO-1 and Claudin-5 in the intestinal epithelial tissues. Results:Compared with the control group, the flunarizine group had lower body mass ratio 1, 3, 5, 7, 10 and 14 d after drug administration (ratio to body mass before drug administration). Compared with the control group, the flunarizine group had significantly shortened residence time in rod rotating and lower rotational speed when falling ( P<0.05). Compared with the control group, the flunarizine group had decreased tyrosine hydroxylase protein in the substantia nigra without significant difference ( P>0.05). Compared with the control group, the flunarizine group had significantly increased interleukin-6 and tumor necrosis factor-α in the substantia nigra (1.00±0.00 vs. 2.79±0.83; 1.00±0.00 vs. 3.39±1.37), significantly lower intestinal Evans blue propulsion rate (80.67%±4.51% vs. 50.67%±6.03%), and statistically decreased ZO-1 and Claudin-5 expressions in the colonic epithelial tissues (27.01±1.41 vs. 16.32±2.83; 37.00±2.80 vs. 24.52±2.12, P<0.05). Totally, 576 microorganisms were noted in both control group and flunarizine group, 744 in the control group alone, and 634 in the flunarizine group alone. The intestinal flora β diversity indices in the 2 groups were significantly different based on weighted Unifrac-principle coordinates analysis (PCoA, PCoA1: 39.88%; PCoA2: 30.69%). Compared with the control group, the microbial colony structure of mice in flunarizine group was dominated by phylum thick-walled bacteria and phylum warty microbacteria, and by families Muribaculaceae, Lachnospiraceae and Akkermansiaceae. Compared with the control group, the flunarizine group had significantly decreased relative abundance of Ackermannia spp. and Lactobacillus spp. in the intestinal flora ( P<0.05). Conclusion:Flunarizine may contribute to the pathogenesis of DIP by causing structural disturbances in the intestinal flora and inducing neuroinflammation based on the gut-brain axis.

Objective:To screen human monoclonal antibodies (mAbs) against the nuclear protein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV), identify their binding specificity to both recombinant NP and NP in viral particles, and determine their affinity constant and binding kinetics.Methods:Antibody genes were extracted from the blood of recovered individuals, and an antibody library was created using phage display. This library was panned by recombinant NP. The selected antibodies were expressed and purified. Enzyme linked immunosorbent assay (ELISA), western blot (WB), and indirect immunofluorescence assay (IFA) were used to assess the binding specificity of these mAbs to recombinant NP and NP in virions. Additionally, biolayer interferometry (BLI) was utilized to determine the antibody affinity constant.Results:An antibody library with a capacity of 7.24×10 7 was successfully constructed. Following three rounds of panning, 6 mAbs (named as NP-1, NP-10, NP-11, NP-20, NP-21, and NP-27) were isolated. The binding specificity of these 6 mAbs against recombinant NP was confirmed through indirect ELISA and WB analysis. Additionally, these mAbs were demonstrated specific in binding to NP in virions as evidenced by IFA detection. The affinity constant values of the 6 mAbs, determined by BLI assay, ranged from 0.47 nmol/L to 32 nmol/L. Conclusions:The 6 mAbs derived from the library are human mAbs that exhibit specificity to the NP of SFTSV and demonstrate a high affinity. These antibodies represent potential candidates for fundamental research and development of diagnostic reagents for SFTSV.

Objective:To screen human monoclonal antibodies (mAbs) against the nuclear protein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV), identify their binding specificity to both recombinant NP and NP in viral particles, and determine their affinity constant and binding kinetics.Methods:Antibody genes were extracted from the blood of recovered individuals, and an antibody library was created using phage display. This library was panned by recombinant NP. The selected antibodies were expressed and purified. Enzyme linked immunosorbent assay (ELISA), western blot (WB), and indirect immunofluorescence assay (IFA) were used to assess the binding specificity of these mAbs to recombinant NP and NP in virions. Additionally, biolayer interferometry (BLI) was utilized to determine the antibody affinity constant.Results:An antibody library with a capacity of 7.24×10 7 was successfully constructed. Following three rounds of panning, 6 mAbs (named as NP-1, NP-10, NP-11, NP-20, NP-21, and NP-27) were isolated. The binding specificity of these 6 mAbs against recombinant NP was confirmed through indirect ELISA and WB analysis. Additionally, these mAbs were demonstrated specific in binding to NP in virions as evidenced by IFA detection. The affinity constant values of the 6 mAbs, determined by BLI assay, ranged from 0.47 nmol/L to 32 nmol/L. Conclusions:The 6 mAbs derived from the library are human mAbs that exhibit specificity to the NP of SFTSV and demonstrate a high affinity. These antibodies represent potential candidates for fundamental research and development of diagnostic reagents for SFTSV.