Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

[摘 要] 目的：通过对比免疫相关不良反应（irAE）发生前后血常规中主要指标的变化，为鉴别诊断irAE及感染性炎症提供新依据。方法：回顾性分析201例2018年8月至2022年6月在河南省肿瘤医院接受抗PD-1抗体治疗后出现irAE的肿瘤患者的临床资料，包括抗PD-1抗体治疗前、发生irAE前及irAE后血常规的主要指标，采用配对t检验分析治疗前后血常规指标值的统计学差异。采用定性变量的配对c2检验分析治疗前后血常规指标值的阳性率（高于正常值的比例）的统计学差异。结果：从201例患者中观察到了258次irAE，其中27例（13.4%）患者发生了2种及以上类型的irAE，214次（82.94%）irAE未引起发热；irAE发生后与抗PD-1抗体治疗前相比，白细胞计数（t=1.087, P=0.278）、中性粒细胞计数（t=0.959, P=0.338）及中性粒细胞百分比（t=0.817，P=0.414）未见明显升高，且三指标高于正常值的病例数分别为28 vs 38（χ2=1.737，P=0.187）、32 vs 44（χ2=2.222，P=0.136）、45 vs 55（χ2=1.240，P=0.265）,差异均无统计学意义。结论：irAE发生后患者外周血白细胞计数、中性粒细胞计数及中性粒细胞百分比无明显变化，这对鉴别诊断感染性炎症可能具有参考意义。

Objective:To investigate the ability of the anti-PD-1(scFv)/hIL-15 fusion protein(PD-15) to specifically bind to PD-1 in vitro and the effect of the combination of PD-15 with GF-hIL-15Ra-K562(G15Ra-K562) feeder cells to expand NK/T cells. Methods:Overlap PCR was used to construct G15Ra expression vector. pMXs-G15Ra-IP was transfected into K562 by electroporation. G15Ra-K562 feeder cell lines were obtained by limiting dilution method. pUC57-PD-15 was constructed by digestion and ligation. Lipofectamine? 2000 was used to transiently transfect pUC57-PD-15 into HEK293T cells and the conditioned medium containing PD-15 fusion protein was obtained. Density gradient centrifugation was used to obtain human peripheral blood mononuclear lymphocytes(PBMC), and CFSE staining was used to mark active proliferating cells. Flow cytometry was used to detect the ability of PD-15 to specifically bind to PD-1 and its effect on the proliferation of human PBMC and the proportion of different subpopulations of lymphocytes.Results:The feeder cells G15Ra-K562 with high expression of fusion protein G15Ra was successfully constructed. The addition of hIL-15 can increase the ability of G15Ra-K562 to expand human PBMC by more than 5 times( P<0.05). PD-15 fusion protein has PD-1 specific binding ability( P<0.001), combined with G15Ra-K562 can efficiently expand human peripheral blood-derived NK/T cells in vitro( P<0.05). The cells expanded by PD-15 and G15Ra-K562 are mainly natural killing cell, CD8 + T and CD4 + T cells. Conclusions:The PD-15 fusion protein can specifically target the PD-1 molecule and has a strong human peripheral blood-derived NK/T cell expansion ability when combined with G15Ra-K562 feeder cells. These results shed light on selective expansion of PD-1 + lymphocytes in vitro.

Objective: To investigate the function of anti-PD-1 (scFv)/IL-15/IL-15Rα-sushi fusion protein (PD-S15) to specifically bind to PD-1 in vitro and to explore its effect on NK/T cell proliferation. Methods: The human anti-PD-1 (scFv) gene sequence and human IL-15/IL-15Rα-sushi fusion gene sequence were synthesized chemically. The recombinant expression plasmid pUC57-PD-S15 was constructed by enzyme digestion and ligation of the two target genes, and then transiently transfected into HEK293T cells by lipofectamineTM 2000. The supernatants of cell culture medium were acquired, and the expression of PD-S15 fusion protein in cell culture supernatants was detected by Wb assay. PBMCs and TILs were cultured in mediums with different proportion of PD-S15/X-VIVOTM15, respectively. Then, the capacity of PD-S15 fusion protein to bind to PD-1 in vitro and its effect on the proliferation of PBMCs and the proportion of CD3+CD8+, CD3+CD4+ and CD3-CD56+ subsets were detected by flow cytometry. The effect of PD-S15 fusion protein on the proliferation of TILs was detected by cytometry. Results: The successful construction of pUC57-PD-S15 eukaryotic expression plasmid was confirmed by double enzyme digestion and sequencing, and then successfully transfected into HEK293T cells. The relative molecular weight of the target protein was approximately 55 000, and was in line with expectations. PD-S15 fusion protein could specifically combine with PD-1 in vitro (P<0.05) and stimulate NK/T cell proliferation (P<0.05). Compared with classical TILs culture method, the efficiency of activation and amplification of T cells in vitro by PD-S15 culturemethodwasbetter (P<0.01). Conclusion: PD-S15 fusion protein can specifically target PD-1 and rapidly expand NK/T cells in vitro, which lays a foundation for the selective expansion of CD8+PD-1+ antigen-specific T lymphocytes from tumor tissues and even peripheral blood.

With the development of tumor precise immunotherapy,it is a hot topic to find biomarkers to predict the response ability of programmed death receptor-1 (PD-1)/programmed death ligand-1 (PD-L1).So far,many predictors,such as the PD-L1 expression in tumor tissue,tumor-infiltrating lymphocyte,tumor mutational burden,serum markers and radiographic markers,have shown predictive value in the process of anti-PD-1/PD-L1 immunotherapy.But each predictor has its limitations.

Objective To explore the influence of AKT inhibitors on tumor infiltrating T lymphocytes (TIL)in patients with liver metastasis of colorectal cancer.Methods The tumor tissues from the patients with liv-er metastasis of colorectal cancer in Department of General Surgery,The Affiliated Cancer Hospital of Zhengzhou University from January 2016 to December 2016 were collected.TIL and tumor cells were isolated by percoll densi-ty gradient centrifugation. The profiling of AKT inhibitors on TIL were analyzed by flow cytometry. Results AKT inhibition enhances the expansion of TIL with memory cell without affects its proliferation,also the cells obtained under AKT inhibitor with IL-2 showed higher frequency of IFN-γproducing cells than IL-2. Conclusion Add AKT inhibitors in TIL cultivation system can strengthen the proliferation of central memory T cells,and does not affect the number of CD8+T cells.This might be developed for cell-based immunotherapy of cancer.

Chimeric antigen receptor modified T (CAR-T) cell therapy has achieved excellent clinical efficacy in patients with hematological malignancies (especially for patients with CD19 positive), and is regarded as a major advance in cancer therapy in recent years. It aroused scientists’strong interest in developing CAR-T cell products for the treatment of cancers. However, there are still some problems in the treatment of CAR-T cells. For examples, some patients lose the opportunity of CAR-T cell therapy while waiting for CAR-T cell culture, some unique adverse events during treatment of CAR-T cell therapy may endanger the patients’life, and the efficacy of CAR-T cell therapy is unsatisfactory on solid tumors. Even for hematological malignancies, some patients will eventually relapse and lead to treatment failure. Therefore, exploring methods to improve the efficacy, diagnosis the unique adverse events of CAR-T cell therapy early and give appropriately management, expand potentially benefiting populations of CAR-T cell therapy are issues that need to be addressed in current CAR-T cell therapy research.

Objective To investigate the effect of Wushen decoction on levels of serum cardiac troponin Ⅰ (cTnI),cardiac muscle enzyme and clinical parameters in patients with sepsis heart failure,and to analyze the correlations between cTnI and myocardial enzyme level and clinical parameters.Methods Forty-two patients diagnosed as sepsis admitted to Wuhan Hospital of Traditional Chinese Medicine from March 2014 to March 2016 were enrolled,and they were divided into a Wushen decoction treatment group and a control group by principle of single blind complete randomized method,21 cases in each group.The patients in control gToup were treated by conventional western medicine,while the patients in Wushen decoction treatment group,on the basis of conventional western medicine,they were treated additionally by Wushen decoction (composed of ginseng,radix sophorae flavescentis,radix glehniae,radix adenophorae,salvia,astragalus,notoginseng radix,rosewood,etc.),one dose a day,the therapeutic course in both groups being 7 days.The changes of biochemical indicators [cTnI,creatine kinase (CK),CK isoenzyme (CK-MB)],haemodynamics parameters [cardiac index (CI),central venous pressure (CVP),extravascular lung water index (ELWI),global ejection fraction (GEF),mean arterial pressure (MAP),heart rate (HR)],treatment condition and prognostic parameters [vasoactive drug dosage index,acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score,duration of mechanical ventilation,the length of stay in intensive care unit (ICU) and total hospitalization time] were compared before and after treatment for 7 days in the two groups.The correlations between the level of cTnI on admission before treatment and CK,CK-MB,APACHE Ⅱ,vasoactive drug dosage index,duration of mechanical ventilation,the length of stay in ICU and total hospitalization time were analyzed.Results The levels of cTnI,CK,CK-MB,CVP,ELWI,HR,vasoactive drug dosage index,APACHE Ⅱ score in two groups after treatment were obviously lower than those before treatment,the levels of CI,GEF,MAP were markedly higher than those before treatment,the duration of mechanical ventilation,the length of stay in ICU and total hospitalization time were significantly shorter than those before treatment,and the changes of above indexes were more remarkable in Wushen decoction group than those in control group [cTnI (mg/L):0.94-± 0.29 vs.1.30 ± 0.67,CK (U/L):96.00 ± 24.30 vs.101.38 ± 24.55,CK-MB (U/L):31.14 ± 6.78 vs.36.48 ± 8.17,CI (mL· s-1 · m-2):64.51 ± 5.83 vs.53.34 ± 4.67,CVP (cmH2O,1 cmH2O =0.098 kPa):10.56 ± 1.84 vs.11.94--2.16,ELWI (mL/kg):8.81±1.61 vs.11.66±2.30,GEF:(33.62±3.88)％ vs.(27.14±4.55)％,MAP (mmHg,1 mmHg =0.133 kPa):84.67 ± 5.58 vs.79.52 ± 5.74,HR (bpm):87.86 ± 9.02 vs.82.95 ± 5.26,vasoactive drug dosage index:2.44 ± 0.53 vs.2.89 ± 0.68,APACHE Ⅱ score:10.66 ± 1.66 vs.14.43 ± 1.82,duration of mechanical ventilation (days):1.67 ± 2.11 vs.2.10 ± 2.26,the length of stay in ICU (days):8.86 ± 2.59 vs.10.67 ± 2.96,total hospitalization time (days):13.24 ± 4.53vs.16.76 ± 5.04,all P ＜ 0.05].On admission before treatment,the correlations between the level of cTnI and CK,APACHE Ⅱ score,vasoactive drug dosage index,duration of mechanical ventilation and the length of stay in ICU were all positive (r =0.322,0.335,0.327,0.328,0.338,P =0.038,0.030,0.030,0.034,0.029).Conclusions The elevation of cTnI level may reflect the degree of myocardial damage in patients with sepsis cardiac failure,and it can be used as an indicator to predict the prognosis of the disease;the changes of many biochemical and clinical indexes suggest that the addition of Wushen decoction might elevate the clinical efficacy for treatment of patients with sepsis heart failure.

Adoptive cell transfusion based on chimeric antigen receptor (CAR) is a new approach for treating malignant diseases and even advanced malignancies. More patients with advanced malignancies are expected to benefit from CAR T-cells once the suitable target molecules are selected, the safe method of gene transduction is applied, and the side effects of CAR T-cells are prevented.