Objective: To establish a rapid, accurate, and decentralized workflow for bacterial whole-genome sequencing (WGS) and risk profiling within the shelf-life of platelet concentrates, and to characterize the species, virulence, antimicrobial resistance (AMR), and immune evasion mechanisms of co-existing bacteria in qualified platelet products, thereby providing a scientific basis for transfusion safety assessment. Methods: Three units of platelet concentrates, which tested negative by routine bacterial screening, were collected from the Chengdu Blood Center between May and June 2025. Samples were enriched at 37℃under six aerobic and nine anaerobic conditions for 7 days. Using a culturomics strategy, aliquots were plated for isolation on days 1, 3, 5, and 7 to obtain cultivable isolates, with negative culture controls included to exclude contamination. High-molecular-weight genomic DNA was extracted via mechanical grinding, purified, and size-selected. Sequencing libraries were constructed and sequenced on the G-seq500 single-molecule nanopore sequencing platform. Genomes were assembled using Flye and polished with NextPolish, with quality evaluated by BUSCO and CheckM. Taxonomic identification was performed using GTDB-Tk. Functional annotation and database comparisons were conducted to analyze virulence factors, AMR genes, and genes related to immune evasion and environmental adaptation. Results: Viable bacteria were successfully isolated from all three qualified platelet units within their shelf life. The isolates were identified as Bacillus albus, Niallia taxi, and Staphylococcus warneri. Nanopore sequencing generated 92 227-109 813 reads (totaling 680-758 Mb) with an N50 of 7 625-8 584 bp and Q20/Q30 scores of 97%/93%, respectively. All three genomes were assembled into complete circular chromosomes with 1-3 plasmids, achieving >93% completeness. Functional analysis revealed that B. albus carried multiple hemolysins, metalloproteases, and multidrug resistance genes, indicating the highest potential pathogenicity and AMR risk. S. warneri exhibited a typical multidrug resistance profile and regulatory network characteristic of coagulase-negative staphylococci, suggesting intermediate virulence. N. taxi harbored few canonical virulence factors and lacked functional AMR determinants, presenting a "low-virulence, low-resistance" profile. Notably, all three strains were enriched in genes encoding antimicrobial peptide resistance systems (e.g., dltABCD, mprF, GraRS, BceAB) and antioxidant enzymes, suggesting a strong capacity to withstand immune stress in the blood environment. Conclusion: Viable bacteria can be recovered from qualified platelet concentrates that test negative by routine screening. Nanopore WGS enables rapid strain-level identification and comprehensive risk profiling of virulence, resistance, and immune adaptation traits. The functional repertoires of these "co-existing" isolates range from environmental adaptation to potential pathogenicity, representing an underappreciated risk for transfusion-transmitted infections in susceptible recipients.

BACKGROUND:There are differentially expressed genes in acute intracerebral hemorrhage,which are related to the occurrence and development of intracerebral hemorrhage. OBJECTIVE:To screen differentially expressed genes and key genes in brain tissue of a rat model with acute intracerebral hemorrhage,to validate them through qPCR,and to analyze the relationships between key genes and the neurological function and brain tissue water content after intracerebral hemorrhage. METHODS:Seventy-eight Sprague-Dawley rats were randomly divided into two groups:in intracerebral hemorrhage group,a rat model of acute intracerebral hemorrhage was made using collagenase injection at the right caudate nucleus;and in sham-operated group,rats were injected with equal amount of saline at the same site.RNA was extracted from rat brain tissues of both groups using the TRIzol method and transcriptome sequencing technology was used to identify differentially expressed genes in brain tissues of acute intracerebral hemorrhage,which were then verified by qPCR and analyzed for the relationships between the genes and neurological function and brain tissue water content after intracerebral hemorrhage.And the key genes were analyzed by GO and KEGG functional enrichment analysis in combination with bioinformatics. RESULTS AND CONCLUSION:Ten key genes were identified,including CXCL8,SERPINE1,TFPI2,CXCR4,GDA,KCNQ5,ERICH3,SCN3B,CACNA1E,and CCL20.The contents of GDA,KCNQ5,ERICH3,SCN3B,and CACNA1E in the intracerebral hemorrhage group were lower than those in the sham-operated group(P＜0.05).The contents of CXCL8,SERPINE1,TFPI2,CXCR4 and CCL20 in the intracerebral hemorrhage group were higher than those in the sham-operated group(P＜0.05).The contents of GDA,KCNQ5,ERICH3,SCN3B,and CACNA1E were positively correlated with brain tissue water content and neurologic deficit score(P＜0.05),while the contents of CXCL8,SERPINE1,TFPI2,CXCR4 and CCL20 were negatively correlated with brain tissue water content and neurologic deficit score(P＜0.05).GO analysis indicated that differentially expressed genes were mainly enriched in two biological processes(leukocyte chemotaxis and chemokine-mediated signaling pathways),two cell components(cation channel complexes and ion channel complexes),and two molecular functions(gated channel activity and ion channel activity).KEGG analysis indicated that differentially expressed genes were concentrated in tumor necrosis factor signaling pathway,glutamatergic synapses and GABAergic synapses.To conclude,the differentially expressed genes in intracerebral hemorrhage include CXCL8,SERPINE1,TFPI2,CXCR4,GDA,KCNQ5,ERICH3,SCN3B,CACNA1E,and CCL20,and these genes are related to brain tissue water content and neurological function after intracerebral hemorrhage.These genes are mainly enriched in cell components,binding functions,cellular protrusions,and other related biological functions.

Objective To explore the presence and potential interactions of microbes and bacteriophages in the blood of healthy individuals by employing in-depth bioinformatics mining to analyze the structure and function of the blood microbi-ome ecosystem.Methods Blood plasma samples from 1 600 voluntary blood donors collected at Mianyang Central Blood Station from 2012 to 2018 were subjected to DNA extraction and library construction.High-throughput sequencing was con-ducted using the Illumina HiSeq 4500 platform,followed by extensive bioinformatics analysis.Microbial abundance in blood samples was analyzed using metagenomic analysis software such as Bowtie2,Trimmomatic and Kraken.Subsequent phage-ome analysis included sequence quality control,assembly,identification,clustering and functional annotation using software such as Megahit,geNomad,CheckV and eggNOG-mapper.Phylogenetic trees,species annotation and host analysis and pre-diction for the identified blood bacteriophages were constructed using iTOL,BLAST and PhaBOX software.Results Met-agenomic sequencing identified microbes across 36 phyla,151 orders,338 families,338 genera and 3 757 species in the plasma samples.At the species level,the most abundant species included Bacillus cereus,Lactobacillus murinus,L.johnso-nii,Faecalibacterium prausnitzii,B.thuringiensis,L.reuteri,Cutibacterium acnes,Dietzia sp.JS16-p6b,Mycoplasma hyo-rhinis,M.hyopneumoniae and Staphylococcus aureus.Through phageome analysis,202 viral Operational Taxonomic Units(vOTUs)were identified,revealing 24 types of bacteriophages.Host analysis using the viral host database completed mat-ches for 15 potential bacteriophage hosts,including Stenotrophomonas maltophilia,Rhodoferax lacus,Pseudoalteromonas marina,Thalassotalea loyana,Vibrio alginolyticus,V.tasmaniensis,V.vulnificus,Pseudomonas sp.,Agrobacterium sp.ST15.13/040,Enterococcus gallinarum,Flavobacterium sp.,Thermotoga naphthophila,Chryseobacterium sp.RU33C,L.acidipiscis and Neisseria mucosa.Conclusion The study of the healthy human blood microbiome and phageome reveals the presence of microbes and phages in the blood,which may have profound impacts on human health.

Objective:To explore the causal relationship between vascular endothelial growth factor A (VEGF-A) and inflammatory bowel disease (IBD) using two-sample unidirectional Mendelian randomization (MR) analysis.Methods:Datasets based on genome-wide association studies (GWAS) of 91 inflammation-related proteins and GWAS datasets related to IBD were collected from the UK Biobank and the IEU OpenGWAS Project. With VEGF-A as the exposure factor, single nucleotide polymorphisms (SNP) associated with IBD were screened as genetic instrumental variables. The inverse variance weighted (IVW) approach served as the primary method in the two-sample unidirectional MR analysis was used to examine the potential causal link between VEGF-A and IBD. Cochran's Q test was utilized to detect potential heterogeneity, while the MR-PRESSO method and MR-Egger intercept test were employed to assess horizontal pleiotropy. A leave-one-out analysis was conducted to evaluate the sensitivity of the results. Results:GWAS data were sourced from the UK Biobank and the IEU OpenGWAS Project databases, including a total of 4 355 UC patients and 2 128 CD patients. The IVW results suggested that VEGF-A may play a protective role in the onset of UC after Bonferroni correction ( OR = 0.9993, 95% CI: 0.9985~0.99997, P = 0.0421; OR = 0.9991, 95% CI: 0.9984~0.9998, P = 0.0095), while no evidence of causal relationship with CD was found ( P = 0.5024, P = 0.3150). Subsequent meta-analysis of the MR results indicated that VEGF-A was a protective factor for UC ( OR = 0.9992, 95% CI: 0.9987~0.9997, P = 0.0011), while no causal association with CD was found ( OR = 1.0000, 95% CI: 0.9997~1.0004, P = 0.8352). The results of Cochran's Q test indicated no heterogeneity, the MR-Egger intercept suggested no horizontal pleiotropy, the MR-PRESSO outlier test detected no outliers, and the leave-one-out sensitivity analysis revealed no abnormal SNP, indicating that the causal inference from the Mendelian randomization analysis had a certain level of reliability. Conclusion:Mendelian randomization analysis indicates causal relationship between VEGF-A and reduced risk of UC, but no causal relationship is found between VEGF-A and CD.

Objective:To explore the causal relationship between vascular endothelial growth factor A (VEGF-A) and inflammatory bowel disease (IBD) using two-sample unidirectional Mendelian randomization (MR) analysis.Methods:Datasets based on genome-wide association studies (GWAS) of 91 inflammation-related proteins and GWAS datasets related to IBD were collected from the UK Biobank and the IEU OpenGWAS Project. With VEGF-A as the exposure factor, single nucleotide polymorphisms (SNP) associated with IBD were screened as genetic instrumental variables. The inverse variance weighted (IVW) approach served as the primary method in the two-sample unidirectional MR analysis was used to examine the potential causal link between VEGF-A and IBD. Cochran's Q test was utilized to detect potential heterogeneity, while the MR-PRESSO method and MR-Egger intercept test were employed to assess horizontal pleiotropy. A leave-one-out analysis was conducted to evaluate the sensitivity of the results. Results:GWAS data were sourced from the UK Biobank and the IEU OpenGWAS Project databases, including a total of 4 355 UC patients and 2 128 CD patients. The IVW results suggested that VEGF-A may play a protective role in the onset of UC after Bonferroni correction ( OR = 0.9993, 95% CI: 0.9985~0.99997, P = 0.0421; OR = 0.9991, 95% CI: 0.9984~0.9998, P = 0.0095), while no evidence of causal relationship with CD was found ( P = 0.5024, P = 0.3150). Subsequent meta-analysis of the MR results indicated that VEGF-A was a protective factor for UC ( OR = 0.9992, 95% CI: 0.9987~0.9997, P = 0.0011), while no causal association with CD was found ( OR = 1.0000, 95% CI: 0.9997~1.0004, P = 0.8352). The results of Cochran's Q test indicated no heterogeneity, the MR-Egger intercept suggested no horizontal pleiotropy, the MR-PRESSO outlier test detected no outliers, and the leave-one-out sensitivity analysis revealed no abnormal SNP, indicating that the causal inference from the Mendelian randomization analysis had a certain level of reliability. Conclusion:Mendelian randomization analysis indicates causal relationship between VEGF-A and reduced risk of UC, but no causal relationship is found between VEGF-A and CD.

Humans ; Carcinoma, Non-Small-Cell Lung/therapy* ; Lung Neoplasms/pathology* ; Anaplastic Lymphoma Kinase ; Neoadjuvant Therapy ; Protein Kinase Inhibitors

Objective:To compare the changes of serum calcium level before and after surgical resection in patients with primary hyperparathyroidism.Methods:Two hundred and seventy-one patients with primary hyperparathyroidism were enrolled from Dec 1992 to Dec 2020 in Beijing Jishuitan Hospital. Serum calcium concentrations were measured before operation, 20 min during surgery, then 2 weeks 1-6 months , 7-12 months and 1 year respectively after operation. The baseline data of postoperative serum calcium such as sex, age, other genetic endocrine diseases, osteopathia and urolithiasis were calculated. The generalized estimation equation was used to analyze the changes of serum calcium in different types of patients before and after operation.Results:The most common postoperative hypocalcemia occurred within 2 weeks, and it occurred frequently half a year after surgery. There was no significant difference in blood calcium between male patients ( t=0.875, P=1.000) and patients with bone lesions ( t=0.034, P=3.049) from 1 to 6 months after surgery and 2 weeks after surgery. Blood calcium level in patients aged 15-35 years old from 1 to 6 months ( t=0.239, P=1.000) , from 7 to 12 months ( t=1.380, P=0.935) and 2 weeks after surgery was not statistically different. The change of bone mineral density was correlated with the change of blood calcium after operation ( F=6.895, P=0.004). Conclusions:The incidence of hypocalcemia was the highest in patients with hyperparathyroidism 2 weeks after surgery, and the blood calcium level was stable within the normal range 1 year later. The blood calcium value of male patients was still at a lower level than that of female patients within six months after surgery. In patients with bone disease, the blood calcium value was lower and recovered slowly 2 weeks after surgery. The blood calcium value of patients aged 15-35 was at a low level within 1 year after surgery.

Sijunzi decoction (SJZD) is a Chinese classical formula to treat spleen qi deficiency syndrome (SQDS) and has been widely used for thousands of years. However, the quality control (QC) standards of SJZD are insufficient. Chinmedomics has been designed to discover and verify bioactive compounds of a variety of formula rapidly. In this study, we used Chinmedomics to evaluate the SJZD's efficacy against SQDS to discover the potential quality-markers (q-markers) for QC. A total of 56 compounds in SJZD were characterized in vitro, and 23 compounds were discovered in vivo. A total of 58 biomarkers were related to SQDS, and SJZD can adjust a large proportion of marker metabolites to normal level and then regulate the metabolic profile to the health status. A total of 10 constituents were absorbed as effective ingredients that were associated with overall efficacy. We preliminarily determined malonyl-ginsenoside Rb2 and ginsenoside Ro as the q-markers of ginseng; dehydrotumulosic acid and dihydroxy lanostene-triene-21-acid as the q-markers of poria; glycyrrhizic acid, isoglabrolide, and glycyrrhetnic acid as the q-markers of licorice; and 2-atractylenolide as the q-marker of macrocephala. According to the discovery of the SJZD q-markers, we can establish the quality standard that is related to efficacy.

Objective:To investigate the effects of early applying of basic fibroblast growth factor (bFGF) on corneal haze formation after surface ablation surgery in rabbits.Methods:The right eyes of 60 healthy New Zealand white rabbits received photorefractive keratectomy (PRK) and were randomized into PRK+ normal saline group, PRK+ bFGF group and simple PRK group, with 20 rabbits in each group.Normal saline solution and bFGF were topically administered according to grouping, respectively, 3 times per day, 1 drop for each time until the sacrifice of the animals, and no drug was used in the PRK group.Another 8 normal rabbits were served as blank control group.The corneal healing response and haze formation were evaluated by anterior segment photography and anterior segment optical coherence tomography (AS-OCT) and graded based on Fantes criteria.Corneal histopathology was examined by hematoxylin-eosin staining.Immunohistochemistry was used to detect the expression of transforming growth factor-β 1(TGF-β 1), α-smooth muscle actin (α-SMA) and matrix metalloproteinase-2 (MMP-2) in cornea.This study protocol was approved by the Experimental Animal Ethic Committee of Affiliated Hospital of Binzhou Medical University (20180209-03). The use and care of the animals complied with the Statement of ARVO. Results:The corneal epithelium was completely healed in 3-4 days following surgery and there was not significantly different in healing time among the three groups.( F=0.57, P=0.57). The haze grading was significantly different among different groups at different time points ( Fgroup=41.736, P<0.01; Ftime=129.445, P<0.01) and showed the highest score in the PRK+ bFGF group on the 28th day after operation.On the 7th day after surgery, AS-OCT image showed that the surface reflection of corneal epithelium was continuous and smooth and corneal epithelium was not tightly attached to the superficial stromal layer; the reflection of the superficial stromal layer was enhanced in all the operation groups.The proliferation of corneal epithelial cells and superficial stromal layer in the operation area were seen under the optical microscope, and the arrangement of collagen fibers in the stromal layer was disordered with the most obvious changes in the PRK+ bFGF group in comparison with the PRK+ normal saline group and the simple PRK group, and these findings became worse on postoperative 28 days.The corneal epithelial surface reflection in the blank control group was continuous and smooth.Immunohistochemistry showed that a few MMP-2 positive cells were seen in the blank control group.TGF-β 1, α-SMA and MMP-2 proteins were positively expressed in the corneas 7 days after surgery in the three groups, and their expressions were the most obvious in the PRK+ bFGF in comparison with the PRK+ normal saline group and the PRK group and were enhanced 28 days after operation, showing statistically differences (all at P<0.05). Conclusions:Early application of bFGF following surface ablation surgery promotes the proliferation of corneal epithelial cells and irregular arrangement of collagen in the superficial stromal layer, which is associated with the expressions of haze-related factors TGF-β 1, α-SMA and MMP-2 in corneas.

Objective:To investigate the association between the pathogenesis of hepatitis B e antigen (HBeAg)-positive chronic hepatitis B and the frequency and function of plasmacytoid dendritic cell (pDC) in patients HBeAg-positive chronic hepatitis B virus infection.Methods:A total of 49 HBeAg (+ ) patients with chronic hepatitis B virus infection in immune tolerance phase (IT) and 100 patients in immune clearance phase (IC) were enrolled. The viral serological indicators and liver function were detected. Peripheral venous blood samples were collected. The peripheral blood pDC frequency and the quantitative expression of co-stimulatory molecule CD86 were detected by flow cytometry, and the correlation between the onset of chronic hepatitis B and the frequency and function of pDC was analyzed.Results:In IC group, hepatitis B surface antigen (HBsAg) levels, HBeAg levels and hepatitis B virus (HBV) DNA loads were significantly lower than those in IT group, and alanine aminotransferase (ALT) levels in IC group were significantly higher than that in IT group; pDC% in IC group was significantly lower than that in IT group; CD86 + pDC% and CD86 mean fluorescentintensity (MFI) showed no significant difference between the two groups. In the IC group, the baseline pDC% was negatively correlated with ALT levels, while CD86 + pDC%, CD86MFI, and CD86 antibody binding capacity (ABC) had no remarkable correlation with ALT levels. Conclusions:The frequency of pDC was correlated with the pathogenesis of CHB. The lower the frequency of pDC in patients with CHB, the more prone to hepatitis. Therefore, increasing the frequency of pDC may inhibit the occurrence of hepatitis.