Objective: To analyze the etiologic distribution of difficult crossmatching and the characteristics of red blood cell antibodies, to summarize compatible transfusion strategies based on serological abnormalities, and to evaluate red blood cell transfusion efficacy and changes in bilirubin levels before and after transfusion. Methods: A retrospective analysis was conducted on difficult crossmatching cases handled by the Department of Blood Transfusion of our hospital from January 2024 to December 2025. Cases that met the criteria for difficult crossmatching determination and had complete data were included in the analysis of the causes and antibody distribution. Patients who completed red blood cell transfusion and had complete hemoglobin (Hb) data before and within 24 hours after transfusion were included in the exploratory subgroup analysis of transfusion efficacy; among them, those who also had total bilirubin and indirect bilirubin data before and after the corresponding transfusion event were included in the bilirubin analysis. According to the main serological abnormal manifestations before transfusion and the actual compatible transfusion management pathways, the cases included in the exploratory subgroup analysis were divided into three groups: the alloantibody-related difficult crossmatching group, the autoantibody-related or autoantibody combined with alloantibody-related difficult crossmatching group, and the ABO blood group identification abnormality-related difficult crossmatching group. The Hb levels before and after transfusion, the Hb increase after transfusion of each unit of red blood cells, and the changes in bilirubin levels before and after transfusion were compared among the groups. Results: A total of 322 difficult crossmatching cases met the predefined criteria, of which 287 were included in the analysis of causes and antibody distribution. There were 183 cases of alloantibody-related difficult crossmatching (63.76%), 54 cases related to inconsistency between ABO forward and reverse typing (18.82%), 44 cases related to autoantibodies (15.33%), and 6 cases related to interference from anti-CD38 monoclonal antibody drugs (2.09%). Among the 183 alloantibody-related cases, 79 cases had only a single alloantibody detected, among which antibodies of the Rh blood group system accounted for 49.37% (39/79). Among single alloantibodies, anti-E was detected most frequently, with 35 cases (44.30%), followed by anti-M with 19 cases (24.05%) and anti-Le with 13 cases (16.46%). A total of 109 cases were included in the exploratory subgroup analysis of transfusion efficacy, including 76 cases in the alloantibodyrelated difficult crossmatching group, 17 cases in the autoantibody-related or autoantibody combined with alloantibody-related difficult crossmatching group, and 16 cases in the ABO blood group identification abnormality-related difficult crossmatching group. The sex distribution and history of previous transfusion differed between cases included in and not included in this analysis. The Hb levels after transfusion in all three groups were higher than those before transfusion, and the differences were statistically significant (P<0.05). The Hb increase per unit of red blood cells did not differ significantly among the three groups (P=0.086). Among 73 cases with paired total and indirect bilirubin data corresponding to the transfusion events, the comparison of changes in total bilirubin among the three groups showed no statistically significant difference (P=0.864), and the comparison of changes in indirect bilirubin among the three groups also showed no statistically significant difference (P=0.914). Conclusion: Alloantibody-related factors were the main causes of difficult crossmatching in this study. Among single alloantibodies, Rh blood group system antibodies, especially anti-E, were relatively common. In the included exploratory subgroup, after individualized compatible transfusion based on the causes of difficult crossmatching, antibody specificities, and crossmatching results, Hb levels were higher after transfusion than before transfusion in all three groups. Within the scope of this study, the magnitude of bilirubin changes did not differ significantly among the different types of difficult crossmatching.

AIM:This study aims to investigate the effect of Agaricus blazei extract FA-2-b-β on ferroptosis of Burkitt lymphoma cells and its mechanism.METHODS:Burkitt lymphoma cell lines Raji and CA46 were treated with FA-2-b-β alone and in combination with ferrostatin-1,a ferroptosis inhibitor,or Stattic,a signal transducer and activator of transcription 3(STAT3)inhibitor.Cell viability was assessed using the CCK-8 method,and the half-maximal inhibitory concentration(IC50)of FA-2-b-β was calculated.Flow cytometry was used to detect apoptosis,cell cycle,mitochondrial membrane potential,and reactive oxygen species(ROS).Additionally,malondialdehyde(MDA)and glutathione(GSH)levels were measured using kits.The mRNA and protein expression levels of ferroptosis-related molecules were determined by RT-qPCR and Western blot.RESULTS:The extract FA-2-b-β at different concentrations significantly inhibited the proliferation of Raji and CA46 cells(P＜0.05),promoted their death,regulated cell arrest in G0/G1 phase,and decreased the mitochondrial membrane potential.(2)ROS and MDA levels were significantly increased with different concentrations of the extract FA-2-b-β(P＜0.05),while the GSH content was significantly decreased(P＜0.05).(3)The protein and mRNA levels of signal transducer and activator of transcription 3(STAT3),p-STAT3,and glutathione peroxidase 4(GPX4)were down-regulated at different concentrations of the extract FA-2-b-β.In addition,prostaglandin-endoperoxide synthase 2(PTSG2)and transferrin receptor protein 1(TfR1)protein and mRNA were up-regulated(P＜0.05),while the protein and mRNA levels of solute carrier family 7 member 11(SLC7A11)were not significantly changed.CONCLU-SION:The extract FA-2-b-β can induce ferroptosis in burkitt lymphoma,and the mechanism may be related to the inhibi-tion of STAT3/GPX4 signaling pathway.

AIM:This study aims to investigate the effect of Agaricus blazei extract FA-2-b-β on ferroptosis of Burkitt lymphoma cells and its mechanism.METHODS:Burkitt lymphoma cell lines Raji and CA46 were treated with FA-2-b-β alone and in combination with ferrostatin-1,a ferroptosis inhibitor,or Stattic,a signal transducer and activator of transcription 3(STAT3)inhibitor.Cell viability was assessed using the CCK-8 method,and the half-maximal inhibitory concentration(IC50)of FA-2-b-β was calculated.Flow cytometry was used to detect apoptosis,cell cycle,mitochondrial membrane potential,and reactive oxygen species(ROS).Additionally,malondialdehyde(MDA)and glutathione(GSH)levels were measured using kits.The mRNA and protein expression levels of ferroptosis-related molecules were determined by RT-qPCR and Western blot.RESULTS:The extract FA-2-b-β at different concentrations significantly inhibited the proliferation of Raji and CA46 cells(P＜0.05),promoted their death,regulated cell arrest in G0/G1 phase,and decreased the mitochondrial membrane potential.(2)ROS and MDA levels were significantly increased with different concentrations of the extract FA-2-b-β(P＜0.05),while the GSH content was significantly decreased(P＜0.05).(3)The protein and mRNA levels of signal transducer and activator of transcription 3(STAT3),p-STAT3,and glutathione peroxidase 4(GPX4)were down-regulated at different concentrations of the extract FA-2-b-β.In addition,prostaglandin-endoperoxide synthase 2(PTSG2)and transferrin receptor protein 1(TfR1)protein and mRNA were up-regulated(P＜0.05),while the protein and mRNA levels of solute carrier family 7 member 11(SLC7A11)were not significantly changed.CONCLU-SION:The extract FA-2-b-β can induce ferroptosis in burkitt lymphoma,and the mechanism may be related to the inhibi-tion of STAT3/GPX4 signaling pathway.

Objective:To investigate the association between peripheral immune function status and lung cancer metastasis,and to identify peripheral blood immune biomarkers for'Deficiency of Vital Qi'assessment in lung cancer metastasis.Methods:A retrospective analysis was conducted on peripheral blood immune markers collected before treatment from lung cancer patients admitted into Shanghai Municipal Hospital of Traditional Chinese Medicine,affiliated to Shanghai University of Traditional Chinese Medicine,between March 2023 and April 2025.Patients were categorized into the non-metastatic and the metastatic groups based on the presence of distant metastasis,and the differences in the expressions of immune cells and cytokines between groups were compared.Peripheral blood immune markers with P<0.05 in univariate analysis were incorporated into a multivariate binary logistic regression model to identify independent predictors of lung cancer metastasis.Results:A total of 193 lung cancer patients were included(101 in the non-metastatic group and 92 in the metastatic group).There were no statistically significant differences between the two groups in terms of gender,age,smoking history,drinking history,or pathological type(all P>0.05).Univariate analysis revealed significant differences in multiple immune markers between the non-metastatic and metastatic groups(all P<0.05),including:lymphocyte count,CD3+,CD4+,and CD8+T,CD19+B cells,absolute counts of CD3-CD16+CD56+NK cells,percentages of Treg cells,CD8+CD28+Treg cells,G-MDSC,and CD3-CD16+CD56+dim NK cells,and levels of cytokine IL-1β,IL-6,and IL-10.Binary logistic regression analysis of differential indicators suggested that the percentage of Treg cells and CD8+CD28+Treg cells in peripheral blood were independent predictors of distant metastasis in lung cancer(OR=1.193,95%CI[1.047,1.36],P<0.01;OR=0.978,95%CI[0.957,0.999],P<0.05).Conclusion:Peripheral blood immune dysfunction is the biological basis for'qi deficiency'in lung cancer metastasis.This study quantitatively demonstrates the correlation between peripheral immune function status and lung cancer metastasis,providing empirical evidence for the theories of'qi deficiency and hidden toxicity'and'metastatic state of tumors'.

Based on the concepts of "people-oriented" in traditional Chinese medicine and "tumor-suppression" in modern medicine， we have combed the studies on the spatial and temporal evolution of tumor metastasis and its biological characteristics in different perspectives， and initially proposed the theory of tumor metastasis from the perspective of the dynamic game between the tumor cells and the body's immune system under the theory of the integration of Chinese and Western medicine， that is， the formation of metastasis is the result of the dynamic evolution of the cancer cells and their surrounding environmental factors in the body over time and space. It is believed that the symptomatic manifestation of metastasis is systematic， the triggering factors of metastasis are constant， and the clinical outcome of metastasis is staged. Accordingly， it is proposed to understand the mechanism of metastasis from the perspective of spatial and temporal dynamics， to establish a clinical and pathological model for identifying metastasis， and to reveal the critical point of metastasis， so as to facilitate the change of the research on tumor metastasis from static to dynamic， and provide ideas for the formulation of metastasis prevention and treatment strategies， and the construction of a new system of metastasis prevention and treatment in the clinical tumor field.

Lung cancer is the leading cause of cancer-related deaths worldwide,with metastasis being the primary cause of mortality in lung cancer patients,and its prevention and control efficacy remain limited.In recent years,immunothera-py has emerged as a promising direction for overcoming the bottleneck of metastasis.Macrophages,as essential components of innate immunity,participate in the entire process of tumor initiation and progression.Tumor-associated macrophages(TAMs)represent the most abundant immune population in the tumor microenvironment(TME),displaying both anti-tumor M1-like and pro-tumor M2-like phenotypes.The latter promotes tumor invasion and metastasis,angiogenesis,lymphangiogenesis,immune suppression,and reactivation of dormant disseminated tumor cells(DTCs),thereby facilitating tumor metastasis.In recent years,traditional Chinese medicine(TCM)has shown significant efficacy in inhibiting tumor metastasis and has been extensively validated.It exerts anti-tumor effects by reducing the recruitment of TAMs,inhibiting M2-like polarization,and modulating cytokines and proteins in the TME.This paper reviews the relationship between TAMs and lung cancer metastasis,elucidates the targets and mechanisms of TCM in regulating TAMs to prevent and treat lung cancer metastasis,aiming to pro-vide insights into lung cancer prevention and treatment.

Objective:To evaluate the diagnostic efficacy of plasma exosomal lncNORAD as a tumor marker for bladder cancer (BCa) by examining its expression levels in BCa patients and analyzing the correlation with clinical pathological characteristics.Methods:This retrospective study. 100 BCa patients diagnosed between October 1, 2021, and October 1, 2023, at Tongji Hospital, Tongji University were randomly selected as the patient group (76 males, 24 females, mean age 71±10 years). In the same period, 40 patients with benign urinary system diseases (benign disease group; 33 males, 7 females, mean age 64±9 years) and 70 healthy individuals undergoing routine check-ups (control group; 53 males, 17 females, mean age 45±12 years) were included. Plasma exosomes were extracted using an exosome extraction kit, and morphologically and molecularly were characterized by transmission electron microscopy, nanoparticle tracking analysis, and Western blotting. Total RNA from plasma exosomes was extracted, and the expression level of exosomal lncNORAD was detected by real-time quantitative polymerase chain reaction. In the BCa patient group, patients were divided into high expression and low expression groups based on the median relative expression level of lncNORAD. The χ2 test was used to compare the relationship between lncNORAD expression and clinical pathological characteristics, and the receiver operating characteristic (ROC) curve was used to evaluate its diagnostic efficacy for BCa.Results:The expression level of plasma exosomal lncNORAD in the patient group was significantly higher than that in the control group ( P<0.001) and the benign disease group ( P<0.001). High expression of lncNORAD was closely related to TNM stage ( P=0.001, χ2=10.187), lymph node metastasis ( P=0.003, χ2=8.575), tumor grade ( P=0.006, χ2=7.440), and nuclear matrix protein 22 (NMP22, χ2=5.844) ( P=0.016). ROC analysis showed that the area under the curve (AUC) for diagnosing BCa with plasma exosomal lncNORAD was 0.828 (95% CI 0.772-0.883). The diagnostic efficacy was further improved when exosomal lncNORAD was combined with the conventional marker NMP22 (AUC=0.866, 95% CI 0.815-0.916). Conclusion:The upregulation of lncNORAD in plasma exosomes of BCa patients is associated with multiple clinical pathological characteristics, indicating its potential as a diagnostic biomarker for BCa.

Objective:To investigate the effects of Feiji Formula on the migration and invasion of lung cancer cells,as well as lung metastasis in animal models and explore its possible mechanisms.Methods:TNMplot,TCGA,and DAVID databases were used to analyze the expression of complement-related proteins(CFHR5[complement factor H-related protein 5],MBL2[mannose-binding lectin 2],C9[complement component 9])in lung metastatic tissues and their relationship with immune cell infiltration,as well as related biological processes and signaling pathways.A subcutaneous xenograft mice model was established using 2LL cells.Mice were administered 2 g/mL Feiji Formula decoction(0.2 mL per dose)via oral gavage for 21 days.The effects of Feiji Formula on the incidence of lung metastasis,the number of lung metastatic nodules,and the protein expression of CFHR5/MBL2/C9 in the lung tissues of the model mice were observed.Exosome tracing assay was performed to observe the secretion and uptake of exosomes by CTC-TJH-01 and H1299 cells.Different concentrations of Feiji Formula were applied to treat H1299 and CTC-TJH-01 cells,and its effects on cell viability,invasion,migration,and CFHR5/MBL2/C9 protein expression were detected by CCK-8,scratch healing assay,Transwell assay,and WB method.Results:Network pharmacology analysis showed that CFHR5/MBL2/C9 proteins were highly expressed in lung metastatic tissues(all P<0.05)and were closely related to the complement system involved in immune response regulation.Compared with the control group,the Feiji Formula group demonstrated a significant reduction in the number of lung metastatic nodules(P<0.05).Feiji Formula(0-200 μg/mL)had no significant effect on the viability of H1299 and CTC-TJH-01 cells(both P>0.05).Both CTC-TJH-01 and H1299 cells could secrete and uptake each other's exosomes.Compared with the 0 μg/mL control group,Feiji Formula at concentrations of 50-200 μg/mL significantly inhibited the migration and invasion abilities of H1299 and CTC-TJH-01 cells(P<0.05 or P<0.01),and significantly reduced the protein expression levels of CFHR5 and MBL2(P<0.05 or P<0.01).Notably,the expression level of C9 protein in CTC-TJH-01 cells increased only after treatment with 200 μg/mL Feiji Formula(P<0.05).Conclusion:Feiji Formula can inhibit the migration and invasion of lung cancer cells as well as lung metastasis in model mice by regulating complement-associated proteins CFHR5/MBL2/C9.This effect may be related to exosome-mediated intercellular communication.

Objective Using bioinformatics methods to study the immune infiltration mechanism of Primary Sj?gren's syndrome(pSS)and to explore potential target Chinese medicines,which can provide new directions for the clinical treatment of pSS.Methods Gene expression profile microarray dataset of pSS was downloaded from the GEO database,differential genes were screened using R software,and gene ontology(GO)and gene pathway enrichment(KEGG)enrichment analysis was performed on these differential genes.Protein interaction network analysis of differential genes was performed by applying the STRING database,key genes were screened by using Cytoscape,and ELISA for the verification of key genes expression.Immune infiltration and correlation of immune cells in pSS were calculated by CIBERSORT inverse convolution method in 22.Finally,the herbal prediction of key target genes was performed by using the Coremine Medical database.Results A total of 232 differential genes were obtained,of which 207 were up-regulated and 25 were down-regulated.GO was mainly enriched in:leukocyte mediated immunity,lymphocyte mediated immunity,leukocyte cell-cell adhesion,etc;KEGG was mainly enriched in Hematopoietic cell lineage,Primary immunodeficiency,Intestinal immune network for IgA production,Phagosome,Leishmaniasis.Ten key genes were screened:PTPRC,CD19,LCP2,CCR5 and CD69 etc.The hub genes expression in the pSS is the same as that of GSE40611.Immune infiltration showed that memory B cells,T cells CD4 memory activated,and T cells CD4 na?ve were highly expressed in the pSS.Immune cell correlation analysis showed a positive correlation between initial Monocytes and T cells regulatory(Tregs),a positive correlation between Macrophages M1 and B cell na?ve,and a negative correlation between Plasma cells and T cells CD4 memory activated.COREMINE Medical predicted that Ginseng,Panax notoginseng,Tripterygium wilfordii,Burnet,Magnolia,and Strychni may treat pSS.Conclusion The development and progression of pSS are the results of the combined involvement of multiple genes and pathways.Memory B cells,T cells CD4 memory activated,and T cells CD4 na?ve may promote the development of pSS.The predicted Ginseng,Panax notoginseng,Tripterygium wilfordii,Burnet,Magnolia,Strychni may be used as target herbs for the potential treatment of pSS.

【Objective】 To investigate the clinical characteristics and antibody distribution as well as evaluate the transfusion efficacy in unexpected antibody positive patients. 【Methods】 A total of 12 235 patients from January 1, 2022 to March 31, 2023 who hospitalized in our hospital and applied for blood transfusion were selected, and those with unexpected antibody were included. The clinical data, including gender, age, diagnosis, blood type, history of transfusion and pregnancy were collected for antibody distribution analysis. Patients who received transfusion were grouped according to the DAT results and the components of red blood cells transfused, and the Hb values of each group before and after transfusion were compared. 【Results】 Among12 235 patients, 118 were positive for antibody screening, with a prevalence of 0.96%. The antibodies from Rh system were the most common (27.43%, 48/175), followed by MNS system (8.57%, 15/175) and Lewis system (6.29%, 11/175), mainly anti-E (18.29%, 32/175), anti-M (8.00%, 14/175) and anti-Le^a (5.71%, 10/175). In addition, 62 transfused patients were divided into group A with suspended red blood cell transfusion and group B with washed red blood cell transfusion for positive DAT, and group C for negative DAT. Hb values (g/L) pre- and post-transfusion were 59.19±15.67 vs 77.52±15.09 in group A, 56.35±14.08 vs 74.44±15.63 in group B, 56.00±12.06 vs 75.00±4.73 in group C, respectively. The Hb values of post-transfusion for three groups were all higher than those of pre-transfusion (P<0.05). 【Conclusion】 Anti-E from Rh system is the most common antibody in patients with unexpected antibody. Appropriate red blood cells transfusion with Hb increases by an average of 6-7 g/L per 1 U of red blood cells indicating good transfusion efficacy. For positive DAT patients, transfusion of suspended red blood cell is feasible.