Mutation Analysis of IgVH Gene in B-Cell Chronic Lymphocytic Leukemia

Summary: The variable heavy chain region (VH) genes of 3 untreated patients with B cell chronic lymphocytic leukemia (B CLL) were cloned and analyzed. The VH family used was VH3 11, VH3 72 and VH3 33. More than 2 % difference from the corresponding germline gene was detected in all the 3 obtained potential functional genes (average 16.7). Mutation pattern analysis indicated evidence of antigen selective pressure observed in 1 of 3 cases. Our findings suggested that the tumor cells originate from post GC cells.

Prognostic Significance of Apoptosis Regulators in B-Cell Chronic Lymphocytic Leukemia

Background: High levels of MCL-1 and BCL-2 proteins have been found in Chronic Lymphocytic Leukemia (CLL), and inversely correlated with response to treatment. BCL-2/Bax ratio is the main director of apoptosis in CLL. The study aimed to clarify the prognostic role of MCL-1, BCL-2 and BCL-2/ Bax ratio in B-CLL. Patients & method: Estimation of MCL-1, BCL-2 and Bax expressions by a flow cytometry in 45 B-CLL patients and the prognostic value of these markers were correlated with other well-known established prognostic markers and treatment response. Results: MCL-1 was expressed in 60% of cases while BCL-2 was expressed in 82.2% of cases. MCL-1 expression was significantly high in male gender, short lymphocyte doubling time (LDT), and high expression of CD 38 (p β2M, CD38 expression), low ZAP-70 expression, splenomegaly and higher Rai stage were significantly increased in patients with high expression of BCL-2 (p β2M, high C-D38 expression, low ZAP-70 expression, the poor cytogenetic and splenomegaly in patients with high expression of BCL-2/ Bax ratio (p In conclusion: MCL-1, BCL-2 expressions and BCL-2/Bax ratio could be useful potential predictive and prognostic markers in B-CLL.

Construction,Expression and In Vitro Biological Behaviors of Ig scFv Fragment in Patients with Chronic B Cell Leukemia

The expression vector of SmIg scFv fragment was constructed in patient with B cell chronic lymphocyte leukemia （B-CLL） and expressed in E. coli to obtain scFv fragment, and the effect of the protein on the proliferation of stimulated peripheral blood mononuclear cells （PBMC） was investigated in vitro. Two pairs of primers were designed, and variable region genes of light chain and heavy chain were amplified by PCR respectively from the pGEM-T vectors previously constructed in our laboratory which containing light chain gene or Fd fragment of heavy chain gene. The PCR product was digested, purified and inserted into pHEN2 vector to construct the soluble expression vector pHEN2-scFv. After the induction by IPTG, the scFv protein was identified by SDS- PAGE electrophoresis and purified by Ni-NTA-Chromatography. MTT was used to determine the effect of purified protein on the proliferation of stimulated PBMC in vitro. Plasmid PCR and restriction enzyme digestion of pHEN2-scFv revealed the pHEN2-scFv vector was constructed successfully. Id-scFv protein was expressed in positive clone after induced by IPTG. SDS-PAGE analysis showed that the relative molecular weight of fusion protein was about 30 kD （1 kD= 0. 9921 ku）, which was consistent with the theoretically predicted value. Proliferation of PBMC could be induced by purified Id-scFv. It was suggested that the expression vector of SmIg scFv fragment was constructed successfully, and scFv protein was expressed and secreted from E. coil, which could induce proliferation of PBMC. This may lay an experimental foundation for further research of Id- HSP complex vaccine for B-CLL.

A comparison of flow cytometry detection of minimal residual disease and chimerism kinetics in chronic lymphocytic leukemia patients after allogeneic hematopoietic stem cell transplantation

Determination of minimal residual disease (MRD) remains crucial for the follow-up after therapy in chronic lymphocytic leukemia (CLL) patients. Chimerism was assessed by short tandem repeat (STR)-PCR and single nucleotide polymorphisms (SNP)-PCR, and MRD by a multicolor flow cytometric approach in 12 consecutive patients with CLL after they received allogeneic stem cell transplantation (SCT). Overall, 11 patients achieved MRD flow negativity [10 had full donor chimerism (FDC) and one had mixed chimerism (MC)]. Only one patient remained with MRD flow positivity and displayed MC. Fifty-six samples were concomitantly studied by both chimerism and MRD flow. A significant correlation was observed between MRD flow data and chimerism in both PB and BM by using a mixed effect linear regression (p < 0.001). Flow cytometry approach of MRD can be easily combined with chimerism during the follow-up post-allogeneic SCT. Both techniques appeared complementary for guiding post-transplant immunomodulation.

Curcumin in chronic lymphocytic leukemia——A review

Curcumin is a widely researched natural product and is known to possess anticarcinogenic properties.Chronic lymphocytic leukemia is a type of leukemia that principally affects patients with age higher than 60 years.Since the toxicity of conventional drugs exceeds the benefits of treating this leukemia type,patients are treated only in the advanced symptomatic stages.The current article reviews curcumin,its general actions and targets in cancer,and specifically that of it in chronic lymphocytic leukemia.

Chronic lymphocytic leukemia/small lymphocytic lymphoma complicated with skin Langerhans cell sarcoma:A case report

BACKGROUND Langerhans cell sarcoma(LCS)is a rare malignancy with poor prognosis.LCS and chronic lymphocytic leukemia(CLL)/small lymphocytic lymphoma(SLL)can occur in the same diseased tissues,such as lymph nodes or skin.CASE SUMMARY A 48-year-old female Han Chinese patient was admitted for generalized lymph node enlargement for 6 years and abdominal distension for 1 wk.She was diagnosed with small B-cell lymphoma(stage IV)/CLL(Benet stage B)and received chemotherapy.She started oral ibrutinib in February 2019.She was hospitalized on June 11,2019,and a 1.5 cm×1.5 cm dark-red nodule with ulceration scalp lesion was found.Biopsy revealed LCS but without CLL/SLL.She was diagnosed with CLL/SLL(Binet stage C,Rai stage IV)accompanied by secondary histiocytic sarcomas and skin LCS and received cyclophosphamide,doxorubicin,vincristine,dexamethasone,and etoposide but developed severe cytopenia.She ultimately refused treatments and discharged spontaneously.She died on September 12,2019.The literature review showed that in patients with CLL/SLL,skin lesions of LCS are accompanied by CLL/SLL.This patient was different from the previously reported cases of skin LCS in patients with CLL/SLL.CONCLUSION In this patient,the skin lesion of LCS showed no concomitant CLL/SLL.

Construction,Expression and in vitro Biological Effects of Idiotype Ig Fab Fragment of B-Chronic Lymphocytic Leukemia

The purpose of this study was to construct expression vectors of idiotype （Id） SmIg in patients with B-chronic lymphocytic leukemia and to express them in E.coli to obtain recombinant Id, and to investigate the effect of the protein on the proliferation and secretion of IL-2 and IFN-γ of stimulated peripheral blood mononuclear cells （PBMC） in vitro. Light chain gene and Fd fragment of heavy chain gene were inserted into fd-tet-DOG2 vector to construct fd-tet-DOG2-Fab. Fab gene was further cloned into expression vector pHEN2 to construct the soluble expression vector pHEN2-Fab. After induction by IPTG, Fab protein was purified by Ni-NTA-chromatography. MTT was used to determine the effects of purified protein on the proliferation of stimulated PBMC in vitro and the concentrations of IL-2 and IFN-γ in the culture supernatants were detected by ELISA. The results showed that recombinant pHEN2-Fab expression vector was constructed successfully. Fab protein was expressed in positive clone after induced by IPTG and two specific bands at 24-25 kD position were observed by SDS-PAGE electrophoresis. Proliferation of PBMC could be induced by purified Fab and the concentrations of IL-2 and IFN-γ in culture supernatants were increased significantly af- ter induction. It was suggested that the expression vector of SmIg Fab fragment was constructed suc- cessfully, and expressed and secreted from E. coli. The Fab protein could induce proliferation of PBMC and promote secretion of IL-2 and IFN-γ.

THE ULTRASTRUCTURAL STUDY OF NON－HODGKIN＇S LYMPHOMA CELL，CHRONIC LYMPHOCYTIC AND HAIRY CELL LEUKEMIA

Non-Hodgkin’s lymphoma cell leukemia (NHLCL),chronic lymphocytic leukemia (CLL) and hairy cell leukemia (HLC) are the diseases very similar to each other. The differential diagnosis is very difficult,especially when there are small lymphoid cells in Periphcral blood and bone marrow under light microscope. We have observed 34 cases with electron microscope. The studies were correlated with clinical manifestation, cytology, pathology and immunologic histochemistry. Ultrastructural features strongly indicated the difference in three various diseases, although all the immunologic markers showed B-cell type.It is concluded that electron microscopic examination is of a definite significance in the diaguosis and successful treatment.

A risk score system for stratifying the risk of relapse in B cell acute lymphocytic leukemia patients after allogenic stem cell transplantation

Background:For patients with B cell acute lymphocytic leukemia(B-ALL)who underwent allogeneic stem cell transplantation(allo-SCT),many variables have been demonstrated to be associated with leukemia relapse.In this study,we attempted to establish a risk score system to predict transplant outcomes more precisely in patients with B-ALL after allo-SCT.Methods:A total of 477 patients with B-ALL who underwent allo-SCT at Peking University People’s Hospital from December 2010 to December 2015 were enrolled in this retrospective study.We aimed to evaluate the factors associated with transplant outcomes after allo-SCT,and establish a risk score to identify patients with different probabilities of relapse.The univariate and multivariate analyses were performed with the Cox proportional hazards model with time-dependent variables.Results:All patients achieved neutrophil engraftment,and 95.4%of patients achieved platelet engraftment.The 5-year cumulative incidence of relapse(CIR),overall survival(OS),leukemia-free survival(LFS),and non-relapse mortality were 20.7%,70.4%,65.6%,and 13.9%,respectively.Multivariate analysis showed that patients with positive post-transplantation minimal residual disease(MRD),transplanted beyond the first complete remission(≥CR2),and without chronic graft-versus-host disease(cGVHD)had higher CIR(P<0.001,P=0.004,and P<0.001,respectively)and worse LFS(P<0.001,P=0.017,and P<0.001,respectively),and OS(P<0.001,P=0.009,and P<0.001,respectively)than patients without MRD after transplantation,transplanted in CR1,and with cGVHD.A risk score for predicting relapse was formulated with the three above variables.The 5-year relapse rates were 6.3%,16.6%,55.9%,and 81.8%for patients with scores of 0,1,2,and 3(P<0.001),respectively,while the 5-year LFS and OS values decreased with increasing risk score.Conclusion:This new risk score system might stratify patients with different risks of relapse,which could guide treatment.

Diagnosis of Leukemia Disease Based on Enhanced Virtual Neural Network

White Blood Cell(WBC)cancer or leukemia is one of the serious cancers that threaten the existence of human beings.In spite of its prevalence and serious consequences,it is mostly diagnosed through manual practices.The risks of inappropriate,sub-standard and wrong or biased diagnosis are high in manual methods.So,there is a need exists for automatic diagnosis and classification method that can replace the manual process.Leukemia is mainly classified into acute and chronic types.The current research work proposed a computer-based application to classify the disease.In the feature extraction stage,we use excellent physical properties to improve the diagnostic system’s accuracy,based on Enhanced Color Co-Occurrence Matrix.The study is aimed at identification and classification of chronic lymphocytic leukemia using microscopic images of WBCs based on Enhanced Virtual Neural Network(EVNN)classification.The proposed method achieved optimum accuracy in detection and classification of leukemia from WBC images.Thus,the study results establish the superiority of the proposed method in automated diagnosis of leukemia.The values achieved by the proposed method in terms of sensitivity,specificity,accuracy,and error rate were 97.8%,89.9%,76.6%,and 2.2%,respectively.Furthermore,the system could predict the disease in prior through images,and the probabilities of disease detection are also highly optimistic.

He Jie Tang in the treatment of chronic hepatitis B patients

AIM： To explore the effect of He .lie Tang （decoction for medication） on serum levels of T lymphocyte subsets, NK cell activity and cytokines in chronic hepatitis B patients. METHODS： Eighty-five patients with chronic hepatitis B were divided randomly into two groups. Fifty patients in group I were treated with He .lie Tang （HIT） and 35 patients in group II were treated with combined medication. The levels of T-lymphocyte subsets （CD^3＋, CD^4＋, CD^8＋）, NK cell activity, cytokines （TNF-α, IL-8, sIL-2R） were observed before and after the treatment. Another 20 normal persons served as group 3. RESULTS： The level of CD^4＋ cells and NK cell activity were lower, whereas the level of CD^8＋ cells in patients was higher than that in normal persons （t = 2.685, 3.172, and 2.754 respectively; P〈0.01）. The levels of TNF-α, IL-8, and sIL-2R in chronic hepatitis B patients were higher than those in normal persons （t = 3.526, 3.170, and 2.876 respectively; P〈0.01）. After 6 months of treatment, ALT, AST, and TB levels in the two groups were obviously decreased （t = 3.421, 3.106, and 2.857 respectively; P〈0.01）. The level of CD^4＋ cells and NK cell activity were increased whereas the level of CD^8＋ cells decreased （t = 2.179, 2.423, and 2.677 respectively; P〈0.05） in group I. The levels of TNF-α, IL-8, and sIL- 2R in group I were decreased significantly after the treatment （t = 2.611, 2.275, and 2.480 respectively; P〈0.05） but had no significant difference in groupII after the treatment （t = 1.906, 1.833, and 2.029 respectively; P〉0.05）. The total effective rate had no significant difference between the two groups （X^2 = 2.882, P〉0.05） but the markedly effective rate was significantly different between the two groups （X^2 = 5.340, P〈0.05）. CONCLUSION： HIT is effective in treating chronic hepatitis B. HIT seems to exert its effect by improving the cellular immune function and decreasing inflammatory cytokines in chronic hepatitis B patients. The function of HIT in protecting liver function in the process of eliminating virus needs to be further studied.

Generation of cytotoxic T lymphocytes specific for B-cell acute lymphoblastic leukemia family-shared peptides derived from immunoglobulin heavy chain framework region

Background Immunoglobulin heavy chain variable region （IgHV） is a well-characterized tumor antigen for B-cell malignancies. It can function as a target for T cell-mediated immune response. Clinical trials of IgHV protein vaccines against lymphoma have demonstrated induction of tumor-specific cytotoxic T lymphocyte （CTL） responses. However, complementary determining regions-based individual vaccines have disadvantages for wide clinical application. Although a recent study demonstrated that immunogenic peptides are derived from framework regions （FR） shared among patients with B-cell lymphoma, how to choose the appropriate peptides for each patient is still unsolved. The aim of this study was to investigate whether immunoglobulin heavy chain FR-derived peptides shared in each IgHV family are potential CTL epitopes presented by B-cell acute lymphoblastic leukemia （B-ALL）. Such CTL epitopes might be beneficial to shifting vaccination strategies against B-ALL from individual specificity to family specificity.Methods Seven IgHV gene families were amplified respectively by PCR and sequenced directly from 71 childhood B-ALL cases. Bioinformatics was applied in analyzing characteristics of sequences available and predicting HLA-A^＊0201-restricted CTL epitopes for each IgHV family. An antigen-specific T cell expansion system was used to generate peptide-specific CTLs. The cytotoxicity of CTLs against B-ALL cells was assessed in the lactate dehydrogenase release assay. Results Complete IgHV rearrangements were identified in all of the 71 B-ALL cases. All of 40 sequences available showed ≥98% homology with the nearest germline IgHV genes, indicating IgHV genes in B-ALL of germline nature. Twelve nonapeptides of high HLA-A^＊0201-binding scores were obtained from 26 productive IgHV protein sequences. Ten （83%） of the peptides were located in FR1 and FR3 shared among the corresponding IgHV family. CTLs specific for the peptide QLVQSGAEV located in FR1 （3-11） shared among the IgHV1 family could be successfully generated from peripheral blood mononuclear cells of two HLA-A^＊0201＋ healthy donors in vitro and were capable of killing HLA-matched B-ALL cell clones belonging to the IgHV1 family. Conclusion Anti-B-ALL CTLs against immunoglobulin heavy chain FR-derived peptides have family-specific cytotoxicity.

Relationship between serum HBV DNA level and HBV-specific, nonspecific cytotoxic T lymphocytes and natural killer cells in patients with chronic hepatitis B

Background The response of patients with chronic hepatitis B （CHB） to antiviral therapy against hepatitis B virus （HBV） is related to the base line level of HBV DNA, but the mechanism is not clear. The present study aimed to understand the possible relationship between the level of HBV DNA and HBV-specific, nonspecific cytotoxic T lymphocytes （CTL） and natural killer （NK） cells of CHB patients and the mechanism how the HBV DNA level influences the antiviral therapeutic effect. Methods Totally 100 adult patients with CHB who were positive for HBV DNA, HBeAg and （HLA）-A2 were enrolled into this study. HBV DNA was tested by real time fluorescence quantitative polymerase chain reaction （PCR）. HBV specific and nonspecific CTL and NK cells were tested by flowcytometry. Serum alanine aminotransferase （ALT） and total bilirubin （TBil） were determined for each patient using routine biochemical tests. The 100 cases were assigned to two groups based on their HBV DNA level： group A had 48 cases, their HBV DNA level was 104-105 copies/ml, group B had 52 cases their HBV DNA level was 106-107 copies/ml. HBV specific CTL, nonspecific CTL, NK cells, ALT and TBil of the two groups were compared. Results HBV DNA level of groups A and B was （4.81±0.39） log10 copies/ml and （6.81±0.40） log10 copies/ml, respectively （t=25.32, P 〈0.001）. HBV specific CTL and NK cells of group A were significantly higher than those of group B （P 〈0.001 for both）. Nonspecific CTL of group A was significantly lower than that of group B （P 〈0.01）. ALT and TBil of group A were significantly lower than those of group B （P 〈0.01 and P 〈0.05, respectively）. Conclusions Serum HBV DNA level of patients with CHB is related to HBV specific CTL, nonspecific CTL and NK cells, which might result in inflammatory reaction of liver and cause more damage to liver function. Mechanism of HBV DNA level affecting the efficacy of anti-viral treatment may be related to the levels of HBV specific CTL and NK cells.

Preparation and Characterization for Bispecific Antibodies of Anti-CD3×Anti-Idiotype to BCellLym phocytic Leukem ia

Bispecific antibodies (BsAbs) of anti CD3×anti idiotype (Id) to B cell lymphocytic leukemia (CLL) were prepared by chemical conjugation and direct hybridization technique of hybridoma and hybridoma without screening markers. The specificity of BsAbs from culture supernatants or ascites was assayed by indirect ELISA and indirect immunoflurescence (IF). The results showed that BsAbs could specifically react with homologous serum IgM from patients with B CLL and cells carrying CD3 marker respectively. Cell combination test and LDH assay demonstrated that BsAb significantly increased the conjugate formation between lymphocyte activated kill (LAK) cells and Daudi cells, and enhanced the cytotoxic activity of LAK cells against Daudi cells.

穿心莲内酯对急性B淋巴细胞白血病L1210细胞的凋亡诱导作用研究

目的探究穿心莲内酯(Andrographolide,AND)对急性B淋巴细胞白血病(B-cell acute lymphocytic leukemia,B-ALL)细胞L1210增殖和凋亡的影响。方法体外培养L1210细胞,取对数生长期的细胞,采用Cell Counting Kit-8(CCK-8)法检测AND对L1210细胞活力的影响;Wright-Giemsa染色法观察AND对L1210细胞形态及数量的影响;JC-1染色观察经AND处理后细胞线粒体膜电位的变化;Western blot法检测经AND处理后细胞PI3K、AKT蛋白表达水平。结果AND在一定时间及浓度内能显著降低L1210细胞活力;Wright-Giemsa染色结果显示,细胞经AND作用24 h后,细胞缩小、染色变深且数量明显减少;JC-1染色结果显示,随着AND浓度增大,线粒体膜电位降低,绿色荧光增强;Western blot结果显示,AND可下调PI3K、AKT蛋白表达。结论AND能抑制B-ALL细胞L1210的增殖并诱导其凋亡,其分子机制与PI3K/AKT通路有关。

淋巴样增强结合因子1在B细胞慢性淋巴增殖性疾病中的表达

目的研究淋巴样增强结合因子1(lymphoid enhancer-binding factor 1,LEF1)在B细胞慢性淋巴增殖性疾病(B cell chronic lymphoproliferative disorders,B-CLPD)中的表达情况,探讨在B-CLPD亚型诊断中的价值。方法回顾性分析58例B-CLPD患者骨髓或淋巴结标本和20例对照组标本,采用免疫组化的方法,检测LEF1的表达情况。结果慢性淋巴细胞白血病(chronic lymphocytic leukemia,CLL)20例,LEF1阳性16例,阳性率为80%;套细胞淋巴瘤阳性为1/12;滤泡性淋巴瘤为1/5;边缘区淋巴瘤为0/11;淋巴浆细胞淋巴瘤为0/8;毛细胞白血病0/2;20例非恶性血液病患者未见LEF1表达;LEF1表达在CLL组差异有统计学意义(P=0.000);LEF1的表达和CD200、CLL积分等CLL的相关指标具有相关性(P<0.05)。4例LEF1阴性的CLL患者中,2例+12染色体异常,检出率高于LEF1阳性组(P>0.05)。结论LEF1在慢性淋巴细胞白血病患者的诊断和鉴别诊断中,有较好的灵敏性和特异性,是B-CLPD鉴别诊断的良好指标。

泽布替尼治疗B细胞淋巴瘤的快速卫生技术评估

目的快速评估新型高选择性布鲁顿酪氨酸激酶(BTK)抑制剂泽布替尼在慢性淋巴细胞白血病(CLL)/小淋巴细胞淋巴瘤(SLL)和套细胞淋巴瘤(MCL)治疗中的有效性、安全性和经济性。方法通过检索PubMed、Cochrane Library、中国知网、万方数据库、维普及卫生技术评估(HTA)网站,收集与泽布替尼相关的系统评价/Meta分析、随机对照试验、药物经济学研究和HTA报告,检索时限为建库/网站起至2023年7月。按照纳入与排除标准筛选文献,分别用相应的评估工具对文献质量进行评价。提取文献数据,对数据进行定性描述。结果共纳入5篇文献,其中3篇为随机对照试验,2篇为经济学研究。在有效性方面,与对照组比较,接受泽布替尼治疗的患者无进展生存期显著延长(P<0.05),总缓解率更高(P<0.05),但两组总生存时间的差异无统计学意义(P>0.05)。在安全性方面,泽布替尼相对于第一代BTK抑制剂伊布替尼的心脏不良事件、主要出血事件发生率和因药物不良事件导致的停药率较低,但相对于传统的化学免疫疗法(苯达莫司汀+利妥昔单抗),泽布替尼的出血事件发生风险仍较高。在经济性方面,泽布替尼相较于伊布替尼在复发或难治MCL患者的治疗中更具有经济性。结论泽布替尼在CLL/SLL和MCL患者中展现出良好的有效性和安全性,同时对于复发或难治的MCL患者具备有一定的经济性优势。

弥漫大B细胞型Richter综合征2例并文献复习

Richter综合征(Richter syndrome,RS)指在慢性淋巴细胞白血病(chronic lymphocytic leukemia,CLL)或小淋巴细胞淋巴瘤(small lymphocytic lymphoma,SLL)的背景下,疾病向侵袭性淋巴瘤进展转化的临床过程。RS转化是恶性血液病进展的一种临床表现,患者在出现RS转变时提示预后差。目前,全球范围内还未形成统一的RS治疗标准。本文回顾性分析2例从CLL向弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)转化的RS患者的治疗方案及预后,探讨化疗后采用造血干细胞移植或嵌合抗原受体T细胞(chimeric antigen receptor T cell,CAR-T)免疫治疗等策略治疗RS的可能性,以为临床实践提供参考与借鉴。

综合需求在B淋巴细胞白血病CAR-T治疗患者自我效能感和生命质量间的中介效应

目的分析综合需求在B淋巴细胞白血病嵌合抗原受体T细胞(CAR-T)治疗患者自我效能感和生命质量间的中介效应。方法采取横断面研究法,选取2022年4月-2023年4月我院收治的B淋巴细胞白血病CAR-T治疗患者127例,采用一般资料问卷、一般自我效能感量表、中文版癌症患者综合需求评估量表(CNAT)、中文版癌症患者生命质量测定量表(FACT-G)进行问卷调查,分析综合需求与自我效能感、生命质量间的相关性,建立综合需求在自我效能感和生命质量间的中介模型并验证。结果回收有效问卷127份(97.69%)。127例患者自我效能感、生命质量和综合需求得分分别为(21.91±5.46)分、(68.18±17.23)分和(127.61±23.49)分。Pearson相关性分析结果显示,患者自我效能感与生命质量呈显著正相关(r>0,P<0.05),综合需求与自我效能感、生命质量呈显著负相关(r<0,P<0.05)。自我效能感对综合需求、生命质量有直接预测作用(P<0.05),综合需求在自我效能感和生命质量间具有部分中介作用,中介效应占总效应的24.35%。结论B淋巴细胞白血病CAR-T治疗患者综合需求处于较高水平,且综合需求、生命质量及自我效能感之间具有密切关系,自我效能感能够直接影响患者的生命质量,还可通过综合需求间接影响患者生命质量;临床应针对患者综合需求进行有效干预,影响患者自我效能感对生命质量的作用路径,提升生命质量。

Modulation of B-cell receptor and microenvironment signaling by a guanine exchange factor in B-cell malignancies

Objective： Chronic lymphocytic leukemia （CLL） and mantle cell lymphoma （MCL） cells over-express a guanine exchange factor （GEF）, Rasgrf-1. This GEF increases active Ras as it catalyzes the removal of GDP from Ras so that GTP can bind and activate Ras. This study aims to study the mechanism of action of Rasgrf-1 in B-cell malignancies. Methods： N-terminus truncated Rasgrf-1 variants have a higher GEF activity as compared to the full-length transcript therefore a MCL cell line with stable over-expression of truncated Rasgrf-1 was established. The B-cell receptor （BCR） and chemokine signaling pathways were compared in the Rasgrf-I over-expressing and a control transfected cell line. Results： Cells over-expressing truncated form of Rasgrf-1 have a higher proliferative rate as compared to control transfected cells. BCR was activated by lower concentrations of anti-IgM antibody in Rasgrf-1 over-expressing cells as compared to control cells indicating that these cells are more sensitive to BCR signaling. BCR signaling also phosphorylates Rasgrf-1 that further increases its GEF function and amplifies BCR signaling. This activation of Rasgrf-1 in over-expressing cells resulted in a higher expression of phospho-ERK, AKT, BTK and PKC-alpha as compared to control cells. Besides BCR, Rasgrf-1 over-expressing cells were also more sensitive to microenvironment stimuli as determined by resistance to apoptosis, chemotaxis and ERK pathway activation. Conclusions： This GEF protein sensitizes B-cells to BCR and chemokine mediated signaling and also upregulates a number of other signaling pathways which promotes growth and survival of these cells.