To ensure medical quality and clinical blood transfusion safety,avoid doctor-patient disputes,and safeguard people's health and social stability,expert group of Chinese Expert Consensus on Standardized Management of Blood Transfusion for Outpatients and Emergency Patients in Medical Institutions,based on domestic and foreign clinical blood transfusion management research results and relevant regulatory guidelines,convened experts and scholars in the field of clinical blood transfusion medicine in China to jointly formulate this consensus,in order to guidance and standardize the blood transfusion management of outpatient and emergency patients in medical institutions and ensure the safety of blood use for patients.

Objective To identify key genes and potential therapeutic drugs associated with Alzheimer's disease(AD)based on microglial senescence using bioinformatics analysis,providing new targets and insights for AD diagnosis and treatment.Methods Gene expression datasets related to microglial senescence and AD,specifically GSE62420 and GSE74615,were downloaded from the Gene Expression Omnibus(GEO)database.Differential Expressed Genes(DEGs)were identified using R software.Functional enrichment analysis,including gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis,was performed using the DAVID online tool.The STRING database was used to construct the protein-protein interaction(PPI)network,and the CytoHubba plugin in Cytoscape software was applied to identify hub genes.The differential expression of hub genes was validated using the GSE129296 and GSE208386 datasets.Potential therapeutic drugs targeting these genes were predicted using the DSigDB database.Results A total of 35 DEGs,including 34 upregulated and 1 downregulated gene,were identified from the intersection of the GSE74615 and GSE62420 datasets.GO and KEGG enrichment analyses indicated that these DEGs were significantly involved in gene expression regulation,protein binding,metal ion binding,and the hypoxia-inducible factor 1(HIF-1)signaling pathway.PPI network analysis and CytoHubba screening identified ten hub genes:HIF1A,secreted phosphoprotein 1(SPP1),integrin alpha X(ITGAX),triggering receptor expressed on myeloid cells(TREM2),glycoprotein non-metastatic melanoma protein B(GPNMB),anexelekto receptor tyrosine kinase(AXL),Cystatin F(CST7),interleukin-12B(IL12B)、lipoprteinlipase(LPL)and fatty acid-binding protein 5(FABP5).Validation using the GSE129296 dataset showed that HIF1A,SPP1,ITGAX,TREM2,GPNMB,AXL,CST7 and FABP5 were significantly upregulated in microglia of AD mice,and the differences were statistically significant(t=8.411～29.49,all P<0.05).Further validation using the GSE208386 dataset indicated that LPL,SPP1,AXL and CST7 were significantly upregulated during microglial senescence,and the differences were statistically significant(t=4.755～5.964,all P<0.05).Based on these validation results,SPP1,AXL and CST7 were identified as key genes in AD.Drug prediction analysis using DSigDB revealed that potential therapeutic compounds targeting these genes exhibit anti-aging,anti-tumor,anti-inflammatory and DNA damage repair effects.Conclusion The key genes SPP1,AXL and CST7,identified based on microglial senescence,may serve as potential biomarkers for AD.Furthermore,multiple potential therapeutic drugs were predicted,offering new targets and strategies for AD diagnosis and treatment.

Objective To evaluate the impact of setting a 4-day incubation time on the detection performance of bloodstream infection(BSI)pathogens by the BacTALERT VIRTUO blood culture system(VIRTUO).Methods Retrospectively collecting blood culture(BC)results from Zhongshan Hospital,Fudan University,from June 2021 to December 2023,and comparing the time-to-detection for clinically significant microorganisms.Results A total of 87 052 BC bottles were collected,of which 7 167(8.23%)were positive.95%of aerobic and anaerobic bottles were reported positive at 65.67 h and 91.2 h respectively.The average(median)time-to-detection for Staphylococcus aureus(S.aureus)was 21.76(16.08)h,Klebsiella pneumoniae(KP)was 15.32(11.65)h,Escherichia coli(E.coli)was 16.14(10.87)h,and Candida species was 35.2(28.5)h.Only 213 bottles(0.24%)were reported positive after 4 days of incubation;among them,88 bottles(41.31%)cultured Propionibacterium acnes(P.acnse).Medical chart reviews for BC reported positive after 96 h(4 days)showed no clinical significance.Simulated growth experiments were conducted on 11 fastidious bacteria,and the median detection time was only 10.8 h,indicating that an incubation time of 4 days is sufficient for fastidious bacteria.Conclusion A 4-day incubation is adequate for VIRTUO system.Reporting 1 day earlier to the doctors can reduce the positive rate of contaminants and the use of antimicrobial agents.

Objective To investigate the role of syntaxin 4(STX4)in endoplasmic reticulum(ER)-mitochondria corcinoma in ovarian cancer(OC)cells.Methods The differences in STX4 expression in the OC cell line SKOV-3 and normal ovarian epithe-lial cells HOSEpiC were analyzed by Western blot and real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).SKOV-3 cell lines with stable knockdown of STX4 were constructed by lentiviral transfected cells,and the knockdown efficiency was verified by Western blot and qRT-PCR.The wound healing assay examined the changes in SKOV-3 migratory ability after STX4 knockdown,the Transwell assay examined the changes in SKOV-3 invasive ability after STX4 knockdown.The ER-mito-chondrial contact structure changes in SKOV-3 cells were visualized by transmission electron microscopy.Intracellular phospha-tidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)pathway proteins and their phosphor-ylated proteins were measured at the protein level.Results Both Western blot and qRT-PCR results showed that the expression level of STX4 was significantly higher in SKOV-3 cells than in HOSEpiC cells,and the differences were statistically significant(t=7.86,9.27,all P<0.05).The knockdown efficiency was verified by Western blot and qRT-PCR.Subsequently,the results of the wound-healing assay indicated that the migratory ability of SKOV-3 cells was significantly inhibited upon STX4 knockdown.The results of the Transwell test showed that the invasive capacity of SKOV-3 cells was significantly reduced after STX4 knock-down(t=7.56,8.82,all P<0.05).Endoplasmic reticulum-mitochondrial contact structure was reduced in SKOV-3 cells with STX4 knockdown under TEM.Finally,no significant changes after STX4 knockdown in the overall levels of PI3K/AKT/mTOR pathway proteins in SKOV-3 cells(t=2.77,5.58,2.99,all P>0.05),but their phosphorylated proteins were significantlydecreased(t=4.42,5.01,5.58,all P>0.05).Conclusion STX4 promoted OC cell migration,invasion,and the formation of ER-mitochondrial contact structures through the activation of the PI3K/AKT/mTOR pathway through phosphorylation.

Objective To investigate the effects and molecular mechanisms of regenerating islet-derived 3-Alpha(REG3A)on the proliferation,apoptosis and cisplatin(DDP)resistance of ovarian cancer(OC)cells.Methods Cancer tissue and adjacent tissue samples of 97 ovarian cancer patients admitted to Qinghai Fifth People's Hospital from January 2021 to December 2022 were collected.Quantitative reverse transcriptase-mediated PCR(qRT-PCR)and immunohis to chemistry(IHC)were used to detect the expression of REG3A in cancer tissues and adjacent tissues,and the relationship between its expression and the clinical pathological characteristics and prognosis of patients was analyzed.QRT-PCR was used to detect the expression of REG3A in human normal ovarian epithelial cell line OSE and human OC cell lines(ES2,HEY,A2780,and SKOV3).SKOV3 and DDP resistant OC cell lines(SKOV3/DDP)were selected and randomly divided into siNC group and siREG3A group.Cell counting kit-8(CCK-8)was used to detect cell proliferation activity.Determination of cell drug resistance by methylthiazolyl tetrazolium(MTT).Flow cytometry was used to detect proliferation cycle and apoptosis.Immunoblotting was used to detect the expression of resistance,proliferation,apoptosis and phsphatidylinostol 3-kinase(PI3K)/protein kinase B(Akt)pathway related proteins in OC cells.Results The expression of REG3A mRNA in ovarian cancer tissues was higher than that in adjacent tissues(1.46±0.43 vs 0.52±0.11),and the difference was statistically significant(t=20.858,P<0.001).The 5-year survival rate of patients with high expression of REG3A was 24.49%(12/49),which was lower than the 91.67%(44/48)of patients with low expression of REG3A,and the difference was statistically significant(χ2=44.841,P<0.001).The proportion of patients with FIGO stages III-IV,moderate to high differentiation and tumor size>3cm in the high expression group of REG3A was higher than that in the low expression group of REG3A,and the differences were statistically significant(χ2=4.537～9.972,all P<0.05).Compared with OSE,the mRNA and protein expression of REG3A in ES2,HKY,A2780 and SKOV3 increased significantly(t=6.725～30.234,all P<0.01).Compared with the siNC group,the expression of REG3A was downregulated in SKOV3 cells in the siREG3A group(0.23±0.02 vs 0.99±0.06),cell activity decreased at 24,48 and 72 hours of transfection,the proportion of G1 phase cells increased,while the proportion of S and G2 phase cells and apoptosis rate,MDR-1,Cyclin D1 and Cleaved caspase 3 protein expression,p-PI3K/PI3K and p-Akt/Akt ratios were decreased,and the differences were statistically significant(t=6.584～22.730,all P<0.01).After treatment with DDP at concentrations of 1.25,2.5,5,7.5 and 15 μg/ml,compared with the siNC group,the survival rates of SKOV3 and SKOV3/DDP cells in the siREG3A group gradually decreased,and the differences were statistically significant(t=2.888～11.135,all P<0.05).Conclusion REG3A is highly expressed in OC tissues and is associated with malignant progression and poor prognosis of OC.Its up-regulation promotes cancer cell proliferation and DDP resistance by activating the PI3K/Akt pathway,and inhibits apoptosis.

Objective The effects of stachydrine(STA)on the proliferation,apoptosis and radiosensitivity of acute myeloid leukemia(AML)cells by regulating the transcription factor forkhead box protein O3(FOXO3)-forkhead box protein M1(FOXM1)signaling axis.Methods Human AML cells(HL-60)were treated with STA at a concentration of 50～1 600 μmol/L,and the activity of HL-60 cells was detected using the cell counting kit-8(CCK-8)method to screen for the optimal drug concentration;HL-60 cells were separated into Control group,low,medium,and high concentration STA groups(STA-L group,STA-M group,STA-H group),STA+lentivirus transfection control group(STA-H+LV-NC group),and high-concentration STA+FOXO3 overexpression lentiviral group(STA-H+LV-FOXO3 group).5-ethyny1-2'-deoxyuridine(Edu)was applied to detect HL-60 cell proliferation;flow cytometry(FCM)was applied to detect cell apoptosis;cell cloning experiments were applied to detect the radiotherapy sensitivity of cells;Western blot was applied to detect the expression of cell proliferation antigen markers(Ki67),Cyclin D1,Caspase-3,B-cell lymphoma2 assaciated X protein(Bax),FOXO3,and FOXM1 proteins.Results STA concentrations of 100,200 and 400 μmol/L were selected for subsequent experiments.Compared with the control group,the positive rate of Ki67,Cyclin D1,Edu,FOXO3 and FOXM1 expression levels in the STA-L,STA-M,and STA-H groups decreased sequentially(tSTA-L=2.169～5.879,tSTA-M=3.089～11.284,tSTA-H=4.572～11.502),Caspase-3 and Bax expression levels,the apoptosis rate,increased sequentially(tSTA-L=9.171,10.082,20.144;tSTA-M=5.435,7.530,7.450;tSTA-H=4.138,4.159,5.956)and the differences were statistically significant(all P<0.05),respectively.Compared with the STA-H+LV-NC group,the positive rate of Edu and the expression levels of Ki67,Cyclin D1,FOXO3 and FOXM1 were obviously increased in the STA-H+LV-FOXO3 group(t=10.055～16.267),Caspase-3 and Bax expression levels,the apoptosis rate were obviously reduced(t=5.736,5.433,8.933),and the differences were statistically significant(all P<0.05),respectively.The colony formation rate of HL-60 cells in the radiotherapy group and STA+radiotherapy group decreased with the increase of radiotherapy dose,and the differences were statistically significant(F=78.630,137.843,all P<0.05),and the colony formation rate of HL-60 cells in the STA+radiotherapy group was lower than that in the radiotherapy group at the same dose(t=1.480～11.301,all P<0.05).Conclusion Stachydrine inhibits AML cell proliferation,induces apoptosis,and enhances radiotherapy sensitivity by inhibiting the FOXO3-FOXM1 signaling axis.

Objective To explore the possible mechanism of T cell immunoglobulin and mucin domain-containing protein-3(TIM-3)regulating M2 macrophage polarization through Galectin-3 in lung injury induced by Streptococcals pneumoniae pneumonia(SPP)in mice.Methods Thirty mice were divided into control group,SPP group and SPP+TIM-3 inhibition group,with 10 mice in each group.The macrophage cell line RAW 264.7 was cultured and divided into a control group,with the SPP group versus the SPP+TIM-3 inhibition group.Lung histopathology was detected by hematoxylin-eosin(H&E)staining.Serum and cell supernatant tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-6 content were measured by enzyme-linked immunosorbent assay(ELISA).Western blotting the protein levels of TIM-3 and Galectin-3 were determined in lung tissues and macrophages.Cluster of differentiation(CD)206,CD86 mRNA levels in lung tissue and macrophages were determined by real time quantitative PCR(RT-qPCR).Results Compared with the control group,the SPP group had significantly abnormal lung tissue pathological structure,thickened alveolar septum,a large number of inflammatory cells infiltration,a large amount of exudate in alveolar space,and increased serum inflammatory factors TNF-α,IL-6 and IL-1β,the protein and mRNA expressions of CD86,the marker protein of M1 macrophages,and the protein and mRNA expressions of CD206,the marker protein of M2 macrophages,and the protein expressions of TIM-3 and Galectin-3 in lung tissues were significantly increased in the model group,and the differences were statistically significally(t=8.36～76.30,all P<0.05).Compared with the SPP group,the SPP+TIM-3 inhibition group had improved lung tissue pathological damage,reduced alveolar septum thickness,inflammatory cell infiltration,reduced exudate in alveolar space,decreased serum inflammatory factors TNF-α,IL-6 and IL-1β content,and decreased expression of CD86 protein and mRNA in lung tissue.The expression of CD206 protein and mRNA was increased,the expression of TIM-3 protein was decreased,and the expression of Galectin-3 protein was increased,and the differences were statistically significant(t=10.67～63.32,all P<0.05).Cell experiments were consistent with this result.Conclusion Inhibition of TIM-3 can improve lung injury in SPP mice by down-regulating the expression of Galectin-3 and promoting M2 macrophage polarization.

Objective To investigate the neuroprotective effect of ligustilide(LIG)-mediated phosphatase and tensin homolog(PTEN)-induced putative kinase 1(PINK1)/Parkin pathway on mitophagy in rats with cerebral ischemia-reperfusion injury.Methods 161 male Sprague Dawley(SD)rats were randomly divided into sham operation(Sham)group,model group,LIG low-dose group,LIG high-dose group,mitophagy inhibitor(Mdivi-1)group,LIG high-dose+Mdivi-1 group,and the positive drug Nimodipine(NMDP)group,each with 23 rats.A modified middle cerebral artery wire thrombus method was used to construct a cerebral ischemia/reperfusion model in rats,and the neurobehavioral scores of rats in each group were compared by Longa's five-point scale;the volume of cerebral infarction was detected by 2,3,5-triphenyltetrazolium chloride(TCC)staining,the histopathology and ultrastructure of the hippocampus were examined by hematoxylin-eosin(HE)staining and transmission electron microscope(TEM).And the Na+-K+-Adenosine Triphosphate was measured by enzyme-linked immunosorbent assay(ELISA);double immunofluorescence staining for translocase of the outer membrane of mitochondrion 20(TOMM20)and Microtubule-associated protein 1 light chain 3(LC3)co-localized area percentage.Flow cytometry assay(FCM)to test the level of reactive oxygen(ROS);real-time fluorescence quantitative PCR(qRT-PCR)was used to measure the relative content of mitochondria in hippocampal neurons;and Western blot was performed to test the level of autophagy and the PINK1/Parkin pathway related protein expression.Results Compared with the Sham group,the neurological function score and cerebral infarction volume of the model group were increased,the hippocampal neurons showed pathological damage such as disordered arrangement,nucleolus disappearance and partial shrinkage of the nucleus and plasma,nuclear membrane rupture,swelling,membrane rupture and crista reduction of some mitochondria,a large number of autophagosomes were observed,and the colocalization area percentage of TOMM20 and LC3 was increased.TOMM20 and cytochrome C oxidase subunit IV isoform 1(COX4I1)in hippocampus and selective autophagy adaptor protein 62(p62)protein expression,mitochondrial encoded ATP synthase 6(mt-ATP6)/Ribosomal protein L13(Rpl13)ratio and Na+-K+-ATPase content decreased,while PINK1 and Parkin protein expression,LC3-II/I ratio and ROS relative content increased,and the differences were statistically significant(t=4.602～52.012,all P<0.01).Compared with the model group,the neurological function score,cerebral infarction volume,pathological and ultrastructural damage of hippocampal neurons were significantly improved in the LIG low,high dose and NMDP groups,and the differences were statistically significant(t=4.851～12.525,all P<0.01).The colocalization of TOMM20 and LC3 and the content of Na+-K+-ATPase were increased,while the expression of TOMM20,COX4I1 and p62 proteins and the mt-ATP6/Rpl13 ratio were decreased in the high-dose LIG group.The protein expression of PINK1 and Parkin,LC3-II/I ratio and ROS relative content were increased,and the differences were statistically significant(t=4.087～33.211,all P<0.01).Compared with the LIG high-dose group,the Mdivi-1 and LIG+Mdivi-1 groups had significantly decreased colocalization of TOMM20 and LC3 and Na+-K+-ATPase content,and significantly increased expression of TOMM20,COX4I1 and p62 proteins and mt-ATP6/Rpl13 ratio.The protein expression of PINK1 and Parkin,LC3-II/I ratio and ROS relative content were decreased,and the differences were statistically significant(t=4.008～43.415,all P<0.01).However,the percentage of TOMM20 and LC3 co-localization area,PINK1 and Parkin protein expression,LC3-II/I ratio and Na+-K+-ATPase content in the hippocampus of the LIG+Mdivi-1 group were higher than those of the Mdivi-1 group.The protein expression of COX4I1 and p62,mt-ATP6/Rpl13 ratio and ROS level were lower than those in MDIV-1 group,and the differences were statistically significant(t=3.721～21.513,all P<0.01).Conclusion LIG may activate mitophagy by regulating PINK1/Parkin signaling pathway to protect neurons from cerebral ischemia-reperfusion injury in rats.

Objective To explore the relationship between rs12881240 polymorphism of paired box gene 9(PAX9)gene and tu-mor susceptibility and efficacy of neoadjuvant chemotherapy in breast cancer patients.Methods A total of 167 patients with breast cancer admitted to the Second Affiliated Hospital of Shaanxi University of Chinese Medicine from January 2022 to Janu-ary 2024 were selected as the study group,and another 110 healthy patients who came to the Hospital for physical examination during the same period were selected as the control group.TaqMan-MGB probe method is used for the single nucleotide poly-morphism(SNP)genotyping rs12881240 of PAX9 gene in peripheral blood,analyzed rs12881240 PAX9 interact gene alleles dis-tribution,and evaluated the genetic susceptibility to breast cancer and neoadjuvant chemotherapy complete remission(pathologi-cal complete response,pCR)relationship.Results In the study group,heavy drinking and absolute value of lymphocyte(LYM#)were higher than those in the control group.The RBC count and PLT count were lower than those of the control group,and the difference were statistically significant(χ2/t=-32.371～63.941,all P<0.05).The genotypes of rs12881240 locus of PAX9 gene were in the control group,and the distribution of study group was consistent with Hardy-Weinberg equilibrium law(χ2=1.647,1.286,all P>0.05),which was representative of the population.The distribution frequencies of alleles C and genotypes CC at rs12881240 locus of PAX9 gene in the study group were higher than those in the control group,and the differences were statisti-cally significant(χ2=14.685,10.401,all P<0.05).In the co-dominant model(CC vs CG)of rs12881240,the risk of CC genotype carriers was higher than that of GG genotype carriers(OR=1.537,95%CI:1.274～2.804,P=0.018).In the dominant model(CG+CC vs GG)and recessive model(CG+GG vs CC),rs12881240 polymorphism was associated with the risk of breast cancer(all P<0.05).Taking age,BMI,menstrual status,heavy drinking,RBC count,PLT count,and absolute value of LYM#as correction factors,rs12881240 was associated with the risk of breast cancer under the three genetic models,and the difference was statistically significant(all P<0.05).There were statistically significant differences in pCR rates between CC,GC,GG geno-types and non-PCR rates at rs12881240 sites(χ2=9.012,P=0.011),and GG genotype carriers had higher pCR rate(χ2=9.012,P=0.011).Conclusion Individuals with genotype CC and individuals with genotype CC+CG have a higher risk of breast cancer.rs12881240 polymorphism of PAX9 gene is associated with breast cancer susceptibility and neoadjuvant chemotherapy pCR in patients,and genotype GG carriers have higher pCR rate.

Objective To explore the effect of tumor necrosis faction-α(TNF-α)gene rs1799964 and C-reactive protein(CRP)gene rs2794521 polymorphism and their interaction on the therapeutic effect of dagliaglozin in patients with heart failure.Meth-ods 98 patients with HF who received treatment in Chengdu Wenjiang District People's Hospital from January 2021 to October 2023 were retrospectively selected for the study.According to the different therapeutic effects,the patients were divided into effective group(n=61)and ineffective group(n=37).The laboratory indexes of the effective group and the ineffective group were compared.The polymorphism,genotype and allele distribution of TNF-α gene(rs1799964)and CRP gene(rs2794521)were analyzed.Multiple regression model was used to analyze the risk factors affecting the efficacy of dagaglizin in the treatment of HF.The association between TNF-α gene(rs1799964)and CRP gene(rs2794521)and cardiac function index was analyzed.To analyze the interaction between TNF-α polymorphism(rs1799964)and CRP polymorphism(rs2794521)on the efficacy of daglipzin in the treatment of HF.Results The frequencies of CC genotype,CT genotype and C allele of TNF-α gene(rs1799964)in ineffective group were 24.32%(9/37),54.05%(20/37)and 51.35%(38/74),respectively,which were higher than those in effective group 11.48%(7/61),40.98%(25/61)and 31.97%(39/122),the differences were statistically significant(χ2=7.284,7.256,7213,all P<0.05).The frequencies of CC genotype and C allele of CRP gene(rs2794521)in ineffective group were 24.32%(9/37)and 47.30%(35/74),respectively,which were higher than those in effective group 6.56%(4/61)and 28.69%(35/122),and the differences were statistically significant(χ2=7.578,6.947,all P<0.05).In the dominant mode and in the cumula-tive mode in the association between TNF-α gene(rs1799964)and CRP gene(rs2794521)was statistically significant(all P<0.05).TNF-α gene(rs1799964)CC genotype(β=1.134,95%CI:1.028～1.964),CRP gene(rs2794521)CT genotype(β=1.357,95%CI:1.239～2.154),CC genotype(β=2.017,95%CI:1.674～4.231)were independent risk factors for the efficacy of daglipzin in treating HF(all P<0.05).The association differences between TNF-α gene(rs1799964)and CRP gene(rs2794521)polymorphism and left ventricular end-systolic dimension(LVESd),left ventricular end diastolic dimension(LVEDd),left ventric-ular ejection fractions(LVEF)and stroke volume(SV)in additive and dominant models were statistically significant(all P<0.05).There were multiplicative and additive interactions between TNF-α gene(rs1799964)genotype CC and CRP gene(rs2794521)genotype CC(all P<0.05).Conclusion The TNF-α gene(rs1799964)genotype CC and CRP gene(rs2794521)genotype CC have a multiplicity and additional interaction,and the risk of daglaglizin in treating heart failure is higher when these two factors coexist.