Smudge cells percentage on blood smear is a reliable prognostic marker in chronic lymphocytic leukemia

Abstract Objective We evaluated the relevance of using the smudge cell percentage in the blood smear as a prognostic marker in CLL. Methods In this prospective study, 42 untreated Senegalese patients with CLL were enrolled. The diagnosis was established, based on the peripheral blood count and flow cytometry using the Matutes score. Cytogenetic aberrations, assessed by fluorescence in situ hybridization (FISH), were available for 30 patients, while the immunoglobulin heavy chain genes (IGVH) mutation status was performed by next-generation sequencing (NGS) in 24 patients. The SC percentage was determined in the blood smear, as previously described. Statistical analyses were executed using the GraphPad Prism 8. Results The mean age was 63 years (48 - 85) and the male: female sex ratio was 4.66. A low SC (< 30%) percentage was correlated with Binet stage B/C (p= 0.0009), CD38 expression (p= 0.039), unmutated IGVH status (p= 0.0009) and presence of cytogenetic abnormalities (for del 13q, p= 0.0012, while for other cytogenetic aberrations, p= 0.016). An inverse correlation was found between the SC percentage and the absolute lymphocyte count (r= -0.51) and patients with higher percentage of SCs had a prolonged survival. However, there was no correlation between the SC percentage and age (p= 0.41) or gender (median, 19% for males vs. 20% for females; p= 0.76). Conclusion When less than 30%, the SC was associated with a poor prognosis in CLL. Easy and affordable, the percentage of SCs in a blood smear could be a reliable prognostic marker, accessible to all CLL patients, mainly those in developing countries.

Evaluation of miR-15a, miR-16-1, ZAP-70, Ang-2, and Bcl-2 as potential prognostic biomarkers in chronic lymphocytic leukemia

Chronic lymphocytic leukemia (CLL) is a blood cancer characterized by the accumulation of clonal B-lymphocytes. This study evaluated the mRNA gene expression of miR-15a, miR-16- 1, ZAP-70, and Ang-2 by qPCR, as well as the plasma levels of Bcl-2 by Elisa immunoassay, in CLL patients and healthy controls. Significant differences were observed when comparing patients and controls regarding miR-15a (p < 0.001), miR-16-1 (p < 0.001) mRNA, Ang-2 gene expression, and Bcl-2 plasma levels (p < 0.001). When stratified by risk, differences were maintained with a significantly reduced expression in high-risk patients. A positive correlation was observed between miR-15a and platelets (R2 = 0.340; p = 0.009) as well as between Bcl-2 and leukocytes (R2 = 0.310; p = 0.019). Conversely, negative correlations were observed between ZAP-70 and platelets (R2 = - 0.334; p = 0.011), between miR-15a and lymphocytes (R2 = - 0.376; p = 0.004), as well as between miR-16-and lymphocytes (R2 = - 0.515; p = 0.00004). The data suggest that a reduction in miR-15a and miR-16-1 expressions, in addition to an overexpression of Bcl-2, are associated with the reduction in apoptosis and, consequently, to a longer survival of lymphocytes, thus contributing to lymphocyte accumulation and aggravation of the disease. By contrast, Ang-2 expression was significantly higher in A than in B + C Binet groups. This context leads to the speculation that this biomarker should be investigated in more robust studies within populations with a still relevantly indolent form of the disease in an attempt to identify those patients with a greater potential for an aggravation of the disease

Research progress on the diagnosis and treatment of CAR-T immunotherapy-induced cytokine release and neurotoxic syndrome / 中国肿瘤生物治疗杂志

@#[摘 要] 嵌合抗原受体基因修饰T淋巴细胞（CAR-T）免疫治疗尤其在复发/难治性恶性血液病的临床应用中显示出卓越的疗效，被认为是最有前景的肿瘤治疗方法之一，但是CAR-T免疫治疗中可引发独特的毒性作用，其中细胞因子释放综合征（cytokine release syndrome，CRS）和免疫效应细胞相关神经毒性综合征（immune effector cell-associated neurotoxicity syndrome，ICANS）在临床应用中最为常见，并具有潜在的致死性，使得CRA-T疗法的广泛应用受到一定的限制，因此如何最大程度地利用CAR-T免疫治疗的同时努力减少或预防CRS和ICANS的发生是目前临床研究的重点，深入了解CRS和ICANS的发生机制能为新的预防措施提供思路。早期识别相关危险因素及鉴定生物预测标志物，熟悉其分级与治疗措施，有利于规范CAR-T免疫疗法的临床应用和患者安全管理。本文就CRS和ICANS的危险因素、发生机制、临床表现、分级与治疗和预防措施等方面进行综述，旨在为CAR-T疗法常见毒性作用的规范管理提供帮助。

Effect Of Auditory Distraction On Hand And Foot Reaction Time Among Ageing Malaysian Automobile Drivers

@#The involvement of ageing drivers in traffic accidents were reported but little cited on the severity of auditory driving distractions. Driving distraction contributes to increases in reaction time which can lead to safety traffic risks. Thus, in this study, hand and foot reaction times were measured in response to different distractions within the identical simulated driving route. The task varies in a controlled setting where soundless distractions were present, Comfortable Loudness Level (CLL), Uncomfortable Loudness Level (ULL) auditory distractions, and phone call distraction. Participants were among 40 Malaysian driving license holders consists of 57.5% males and 42.5% females with age mean, (M=51.83, SD=14.058). Results indicated that both hand and foot reaction time were shortest for CLL and longest during phone call. Ageing male scored shortest hand reaction time of 1.15s during CLL distraction. For foot reaction time, ageing male scores shortest of 0.92s for both CLL and no distractions. Pearson’s coefficient of correlation shows r>0.5. The results indicated hand reaction time was affected by foot reaction time (r=0.665), was significantly more for foot when compared with hand, could be because of difference in nerve conduction velocity and movement time of the hand when compared with that of foot.

The current status of anti-GPCR drugs against different cancers / 药物分析学报

G protein coupled receptors (GPCRs) have emerged as the most potential target for a number of drug discovery programs ranging from control of blood pressure, diabetes, cure for genetic diseases to treatment of cancer. A panel of different ligands including hormones, peptides, ions and small molecules is responsible for activation of these receptors. Molecular genetics has identified key GPCRs, whose mutations or altered expressions are linked with tumorgenicity. In this review, we discussed recent advances regarding the involvement of GPCRs in the development of cancers and approaches to manipulating the mechanism behind GPCRs involved tumor growth and metastasis to treat different types of human cancer. This review provides an insight into the current scenario of GPCR-targeted therapy, progress to date and the challenges in the development of anticancer drugs.

Smudge cell percentage as a surrogate marker for ZAP-70 expression in patients with chronic lymphocytic leukemia

BACKGROUND: This study aimed to evaluate the prognostic value of smudge cell percentage as a surrogate marker for zeta-chain-associated protein kinase 70 (ZAP-70) expression in chronic lymphocytic leukemia (CLL) patients. METHODS: Sixty three newly diagnosed CLL patients were investigated at the Hematology Department of the Medical Research Institute of Alexandria University with complete blood count, lactate dehydrogenase, β2 microglobulin levels, ZAP-70 expression, and estimation of the percentage of smudge cells. RESULTS: The percentage of smudge cells ranged from 2 to 58% with a mean of 24.03±13.98%. Higher percentages of smudge cells (>30%) were statistically significantly associated with markers of better prognosis (negative ZAP-70, early-stage disease according to the Binet and Rai staging systems, as well as low and intermediate risk CLL prognostic index). The percentage of smudge cells showed significantly negative correlation with the ZAP-70 expression and higher area under the curve for prediction of ZAP-70 positivity with better survival for 36 months in patients with >30% smudge cells. CONCLUSION: The percentage of smudge cells at presentation of newly diagnosed CLL patients could be used as a surrogate marker for ZAP-70 expression and an additional prognostic marker for disease progression.

Quantitative real time PCR analysis of apoptotic gene expression in chronic lymphocytic leukemia patients and their relationships with chemosensitivity

Background: Apoptosis-related gene expression such as BCL2, and p53 has been suggested in predicting the patient response to chemo- or radiotherapy, as well as patient's survival. Methods: The aim of this study was to determine changes in BCL2 and p53 apoptosis related gene expressions in chronic lymphocytic leukemia (CLL) patients in response to different chemotherapy regimens and number of treatment courses. The study was conducted on 55 CLL patients (44 CLL and 11 CLL/SLL; small lymphocytic lymphoma) and 40 healthy individuals as control, over three-months period. The RNA was extracted by exploitation total RNA extraction kit, treated with DNAse, then cDNA was synthesized and qRT-PCR used to analyze antiapoptotic BCL2 and tumor suppresser p53 gene expressions. Results: CLL/SLL showed higher BCL2 and p53 gene expression than CLL. The patients with CLL showed three-fold increase in BCL2 gene expression compared to healthy controls (p < 0.05), and 50% decrease in p53 gene expressions (p < 0.05). BCL2 gene expression was higher, particularly, for those who were treated with higher range of treatment courses and combination of fludarabine, cyclophosphamide and rituximab (FCR) regimen. P53 gene expression reciprocally related with BCL2 and vice versa. Conclusions: In contrary to BCL2, p53 gene was extremely expressed in patients treated with chemotherapy agents, particularly after 24­30 months disease duration; suggesting a late expression of p53 during advanced stages of the disease. A proportional change in BCL2 and p53 gene expression was reported with different treatment regimens; Chlorambucil (Clb) decreased and FCR regimen increased BCL2 gene expression. Higher p53 gene expression reported with the Chlorambucil + (Chlorambucil + Prednisolone) regimen (AU)

Stabilization of Chronic Lymphatic Leukemia with Amanita phalloides: Effect of Additional Chelidonium majus. Case Report.

Objectives: Amanita phalloides contains amanitin, inhibiting RNA polymerase II. Partial inhibition with amanitin influences tumor cell - but not normal cell - activity. A patient with diagnosed B-cell chronic lymphatic leukemia was treated successfully for eight years with Amanita phalloides. However, the necessary dose for stabilization of the disease increased during this time. In addition thrombocyte levels decreased, indicating bone marrow affection. Therefore additional regimen was necessary. Methods: Chelidonium majus contains alkaloids with cytostatic and cytotoxic potential. In addition to Amanita, Chelidonium was applied. Results: During treatment with Amanita and Chelidonium, leukocyte levels and lactate-dehydrogenase levels were roughly unaffected, meaning there is no strong effect on tumor growth of cells, and no observable cell destruction. Anyhow, thrombocyte levels increased after the Chelidonium treatment, indicating an effect in the bone marrow. Conclusion: Chelidonium majus can be useful during Amanita therapy as a pulse to regenerate bone marrow affection.

Borrelia Infection Appears as Chronic Lymphocytic Leukemia: Therapy with Amanita phalloides and Terebinthina laricina.

Background: To date, Chronic Lymphocytic Leukemia (CLL) has been viewed as a malignant disease with tumor growth of cells. This hypothesis should be reviewed. Methods: A patient diagnosed with CLL is treated with Amanita phalloides, containing amanitin, inhibiting specifically tumor cell activity without affecting normal cells. Despite initial leukocyte cell count decrease, further therapy fails after eight months. He suffers from severe symptoms of inflammation, not specific for CLL. Additional Borrelia infection is diagnosed and Terebinthina laricina is applied. Results: Herxheimer reaction occurs some weeks later, accompanied by continuous leukocyte cell count drop to normal range within four months, even after stopping Amanita therapy. All symptoms of borreliosis and CLL vanished. Conclusion: CLL might be induced by a Borrelia-infection. This should be considered in diagnostic and therapeutic regimen.

Function and Regulation of miR-9-3 in Chronic Lymphocytic Leukemia / 现代检验医学杂志

Objective To investigate the role and regulatory mechanism of micro RNA-9-3 (miR-9-3)in the pathogenesis of chronic lymphocytic leukemia.Methods Using the methylation specific PCR (MSP)technology to detect 8 cases of normal bone marrow tissue and peripheral blood,78 cases of bone marrow tissue came from the chronic lymphocytic leukemia pateints newly diagnosed and the methylation level of 7 kinds of leukemia cell line.Used Western blot to detected the NF-kappa B1 signal transduction pathway activation levels of methylation positive leukemia cell line.Results The miR-9-3 of normal control group were in the negative methylation status.Only I83-E95 and WAC3CD5+ were in positive methylation status in seven kinds of leukemia cell line (the positive of MSP was 28.6%);65 cases occurred miR-9-3 methylated in 78 of chronic lymphocytic leukemia patients (the positive of MSP was 83%).I83-E95 and miR-9-3 cells of WAC3CD5+ were in the methylation state when treatment with 5-nitrogen-2’-deoxidization cytidine (5-Aza2’Dc).Conclusion The abnormal methylation of miR-9-3 were usually seenin chronic lymphocytic leukemia,it could lead to abnormal hyperplasis in cancer cells.The methylation of miR-9-3 could inhibit the activation of NF-kappa B1 signal pathway suggested that it could sup-press the apoptosis of cancer cells through this pathways to trogered the progression of disease.The inhibitor of methylation could be induced the demethylation of leukemia cell lines,so it is possible that miR-9-3 maight be a new gene targets for the treatment of chronic lymphocytic leukemia.

Novel Agents in the Treatment of Chronic Lymphocytic Leukemia / 대한내과학회지

Chronic lymphocytic leukemia (CLL) is an indolent B-cell leukemia, and the most common leukemia in the Western world. While CLL is currently a rare leukemia type in Korea, the prevalence of CLL is expected to rise in the future. The clinical course is extremely heterogeneous and can range from asymptomatic to rapidly progressive disease. Chemoimmunotherapy (rituximab + fludarabine + cyclophosphamide) has become the standard front-line therapy in young fit patients with normal renal function. Despite the development of more intensive new treatments, these new therapies are not suitable for all patients with CLL. Novel targeted therapies, such as small molecules that disrupt the B-cell receptor pathway or new monoclonal antibodies, have emerged as promising therapeutic options. Prospective clinical trials exploring the efficacy and toxicity profiles of new agents, alone or in combination with other treatments, will reveal whether these novel agents could replace the current standard regimens.

Occurrence of chronic lymphocytic leukemia in patients with chronic myelogenous leukemia.

Chronic lymphocytic leukemia (CLL) is the most common leukemia of adults in the western world and constitutes about 33% of all leukemia’s. The incidence of CLL increases with age and are more common in older population. Chronic myeloid leukemia (CML) on the contrary occurs in both young adults and elderly and is a chronic myeloproliferative disease that originates from abnormal pluripotent stem cells and results in involvement of multiple hematopoietic lineages, but predominantly myeloid and less commonly lymphoid. Association between CLL and myeloid malignancies (CML, acute myeloid leukemia and MDS, myelodysplastic syndrome) is rare. In literature documenting CLL and CML in same patients, occur either simultaneously or CML is preceded by CLL.

Clinical Significance of p53 Protein Overexpression and Serum Anti-p53 Antibodies in Patients with Acute and Chronic Leukemia / 대한임상병리학회지

BACKGROUND: Mutations of p53 gene, rarely found in leukemia, result in accumulation of mutated p53 protein in the nuclei of tumor cells, which can be detected by immunohistochemistry. Lately, anti-p53 antibodies were found in the sera of patients who had solid tumors as a result of immune response to accumulation of mutated p53 protein in tumor cells. METHODS: For investigation of the clinical implication of cellular p53 protein overexpression and serum p53 antibody, immunohistochemical staining for p53 protein of B-5 fixed paraffin embedded bone marrow biopsies and enzyme immunoassay for the presence of anti-p53 antibodies of sera were performed simultanously; in 58 cases of AML, 34 cases of ALL, 11 cases of acute leukemia at relapse, 13 cases of CML in chronic phase and 5 cases of CLL. RESULTS: Overexpression of p53 protein was found in 9.1%(11/121) of all leukemias, with 8.6% of AML with predominance of M6, 5.9% of ALL, 18.2% of acute leukemia at relapse and 40% of CLL, but not found in CML. Serum anti-p53 antibodies were found in 5.8%(7/121) of all leukemias, with 6.9% of AML and 5.9% of ALL, 9.1% of acute leukemia at relapse, but not found in chronic leukemias. In AML and ALL, age, sex, hemoglobin, leukocyte count, platelet count and blast % were not related with p53 protein expression. The AML patients with p53 protein overexpression have more unfavorable karyotypes(complex karyotype, -5, -7 and t(10;11)), with shorter overall survival as compared to those without p53 protein overexpression. The presence of serum anti-p53 antibodies was not related with clinical findings of leukemias. CONCLUSIONS: The indications are that p53 gene alterations will contribute to disease development and progression in some specific patients with leukemia, due to the rare frequency of overexpression of p53 protein and serum anti-p53 antibodies in leukemia. Analysis of the p53 protein and serum p53 antibodies could screen p53 gene mutation and predict prognosis for some leukemias.

The changes of functions of chemokine receptor 9 on T cell lymphocytic leukemia patients / 中国免疫学杂志

Objective:To observe the influences of functions of chemokine receptor 9(CCR9) on leukemia patients,we analyzed 38 typical T cell lymphocytic leukemia cases.Methods:The functions of T ALL and T CLL CD4+T cells towards TECK/CCL25 were determined by chemotaxis assay and adhesion assay.Results:Almost in all of the T ALL patients,TECK/CCL25(a ligand for CCR9)could induce a high chemotactic migration of T ALL CD4+ cells as well as a high adhesion.Conclusion:It indicates that CCR9 and its ligands may promote survival or proliferation of T ALL cells. [

A Case of Adenocarcinoma of the Lung In Patient with Chronic Lymphpcoytic Leukemia / 대한암학회지

In Korea chronic lymphocytic leukemia(CLL) is a rare diesase with evidence immunologic incompetence. The immunodeficiency of patients with CLL could place them at increased risk of new cancers. The incidence of secondary malignancy, including lung cancer has increased in patients with CLL and this should be also considered in the differential diagnosis of a new pulmonary infiltrate appearing in patients with known CLL. A-73-year old male patient with CLL was admitted to our hospital because of dyspnea. Chest X-ray showed multiple scattered small nodules in both lung fields and alveolar consolidation mass at right lower lung. Histopathological examination of the bronchoscopic biopsy specimen demonstrated that he had a moderatly differentiated adenacarcinoma of lung. Since we experienced a case of metachronous adenocarcinoma of lung in patient with known CLL, we report this with a review of literatures.