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Background and purpose: So far many protein factors capable of inhibiting the immune system are secreted by different cancer cell lines in the culture supernatant and characterized. Ïn this report, the release of DNÂ fragments into the culture supernatant of human promyelocytic cell line, HL-60, with immunosuppressive activity was investigated.
Materials and methods : HL-60 culture supernatant was concentrated and applied to ion-exchange DËÂË-Sepharose ÇL-6B column. The column was eluted with NaÇl gradient and the fractions were collected for assay. Fractions with immunosuppressive activity were purified with HPLÇ.
Result: The data show that trypsin and RNase have no effect on antiproliferation, but DNase destroyed the immunosuppressive activity. The DNÂ with irreversible immuno-suppressive activity was found to be double-stranded with the molecular weight range between 0.6 and 1.5 kilobases. This DNÂ did not change the cell number and viability and as a result was not cytotoxic. Çell cycle analysis has suggested that DNÂ molecule could arrest the mitogen-stimulated lymphocytes in G1 phase, and prevent the cells from entering into the proliferating S phase.
Çonclusion: These data show that DNÂ like immunosuppressive proteins could inhibit the proliferation of lymphocytes as well as the proliferation of some cancer cells.

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Background and purpose: Üsing immunotyping in classifying leukemia is growing increasingly. This method provides useful information about proper classifying of leukemia, the prognosis and treatment of this disease. This study is conducted to determine frequency percentage of ÇD markers in patients suffering from acute leukemia who had referred to Ïmam Khomeini Hospital in Ürmia for one year.
Materials and methods: The present descriptive study was conducted in Ïmam Khomeini Hospital in Ürmia from 22 July 2008 to 23 July 2009. Random samples were taken from pediatrics and adults’ blood ward from the 119 referred patients within one year. Then, flowcytometry markers were measured, and environmental blood slide and marrow aspiration were studied.
Results: The results demonstrated that 61 of them suffered from acute leukemia, 22 cases had chronic lymphoproliferative, and 36 patients were among miscellaneous group. Ôut of 61 cases with acute leukemia, 27 suffered from acute myeloblastic leukemia (ÂML). Âmong the 27 cases with ÂML, one (3.7%) suffered from M1, 4 cases (14.8%) from M2, 5 cases (18.5%) from M3, 9 cases (33.3%) from M4, and 8 cases (29.6%) from M5. ÇD13 and ÇD33 markers were the most frequent ones in ÂML patients. Ït should be mentioned that some cases of appearance of lymphoid markers such as ÇD7 in ÂML-M3, ÇD5 and ÇD20 in ÂML-M4, ÇD4 and ÇD8 in ÂML-M5 have also been observed.
Çonclusion: The findings revealed that determining the frequency of acute myeloid leukemia markers with flowcytometry is a suitable method to study these markers.

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Background and purpose: Oligodendrocytes are responsible for myelin synthesis in the central nervous system (CNS). Olig1 and Olig2 play an important role in regulating the development of oligodendrocyte precursor cells (OLPs). Myelin basic protein (MBP) is the main component of myelin sheath. Leukemia inhibitory factor (LIF) has an important role in myelination and pathology of multiple sclerosis (MS). This study investigates the effects of LIF on the expression of MBP, Olig1 and Olig2 in the cerebral cortex of cuprizone-induced MS mice. Materials and methods: In order to induce MS the mice were treated by Cuprizone for 5 weeks. The mice were then divided into three groups. The first group was injected LIF (25 µg/kg BW per day, i.p.) for 6 weeks. The second group (SHAM) received normal saline, i.p. for six weeks and the third group did not receive any injection. The mice were killed after six weeks and the cerebral cortex was harvested and MBP, Olig1 and Olig2 expression was studied by western blotting. Results: The results indicate that MBP, Olig1 and Olig2 expression in the cerebral cortical extracts was significantly increased in the LIF injected group compared with control and SHAM groups (P<0.001). Conclusion: This study reveals that LIF not only increases Olig1 and Olig2 expression in the oligodendrocytes and enhances oligodendrocytes activity and MBP expression, but also plays an important role in the pathophysiology of MS.

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Background and purpose: Chronic myeloid leukemia (CML) is the most widely recognized form of leukemia. Nowadays, trying to find new synthetic compounds is one of the basic strategies to find new drugs to treat CML. In present study the effects of some of Pyrimidone derivatives on proliferation, viability, and apoptosis of human leukemia K562 cell line as experimental model for CML have been investigated. Material and Methods: K562 cells were treated by various concentrations of 4 amino pyrimidone derivatives. Anti- proliferative effects of the compounds were studied using trypan blue exclusion test. Apoptosis was detected by fluorescent microscope and DNA fragmentation assay. Results: Our study showed that: 1) aminopyrimidone derivatives induced growth inhibition in K562 cells in concentration- and time-dependent manner, 2) The aminopyrimidone derivatives inhibited viability and induced apoptosis in these cells. 3) The apoptotic and growth inhibitory effects of these compounds were dependent on their molecular structure. Conclusion: According to growth inhibitory and apoptotic effects of new synthetic, aminopyrimidone derivatives could be proposed as new and effective compounds for more investigations in treatment of leukemia.

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Background and purpose: Acute leukemia is a malignancy with a rapidly progressive course of the lymphoid or myeloid cells. A prognostic factor for its survival and treatment is of particular importance in medical research. Therefore, this study aimed at applying competing risk model to compare clinical and pathologic factors in pediatric acute leukemia. Materials and methods: A historical cohort analysis was performed on survival data of 97 children with acute leukemia collected from Pediatric Oncology Department in Booali Sina Hospital in Sari from 2006 to 2014. We recorded data including age at diagnosis, type of leukemia, sex, white blood cell count, and platelet count. Statistical analysis using competing risk model, Cox regression model and STATS software were performed. Results: The median follow-up period was 14.83 mounts during which 27 patients (27.83%) (17 ALL cases and 10 AML cases) passed away. In competing risk model significant relationship was found between type of leukemia, hemoglobin count and white blood cell count with survival time. Conclusion: In order to identify the prognostic risk factors on the survival of patients with acute leukemia, in presence of competing risks, using the competing risk regression model was found more efficient compared to cox regression model.

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Background and purpose: Chronic lymphocytic leukemia (CLL) is characterized by clonal proliferation and accumulation of long-lived B lymphocytes within the peripheral blood, bone marrow, and secondary lymphoid organs. In this study, immunophenotypic characteristics of leukemic cells and their association with disease prognosis were investigated in CLL patients.

Materials and methods: A case-control study was carried out in 25 CLL patients and 15 healthy individuals. Complete blood count was performed for all subjects. CLL patients were classified into different clinical stages based on the Rai staging system. For immunophenotypic characterization of leukemic cells, peripheral blood mononuclear cells (PBMCs) were isolated from CLL patients and stained using specific monoclonal antibodies. They were then analyzed by flow cytometry.

Results: The CD5, CD19, CD20, and CD23 were expressed significantly more in advanced clinical stages of CLL compared to those in primary stages (P= 0.03, 0.01, 0.02, and 0.02, respectively). The mean fluorescence intensity (MFI) of CD38 in advanced progressive stages of CLL patients was higher than that of primary non-progressive stages (P=0.02).

Conclusion: Our results indicated significant immunophenotypic profile and higher CD38 expression in CLL patients with advanced clinical stages. These immunophenotypic characteristics are useful biomarkers for the disease monitoring and therapy.

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Background and purpose: We aimed at evaluating the effectiveness of cognitive rehabilitation on executive functions (attention and working memory) in adolescents surviving acute lymphoblastic leukemia with a history of chemotherapy.

Materials and methods: This clinical trial was conducted in 30 adolescents who survived acute lymphoblastic leukemia and had a history of chemotherapy. The participants were randomly assigned to two groups of intervention and waiting list (n=15 per group). The subjects in the experimental group were provided with 12 45-minute sessions of cognitive rehabilitation of memory and attention. The data was obtained using Wisconsin Card Sorting Test and Continuous Performance Test in the three stages of pre-test, post-test, and follow-up. Data analysis was performed using descriptive (mean and standard deviation) and inferential statistics (MANCOVA).

Results: Wisconsin test showed significant improvements in the scores of working memory and shifting attention in components of correct responses, errors, classes, and preservation variable in the intervention group (P<0.05). Moreover, significant performance enhancement was observed in the continuous attention test and components of commission error and error of omission in continuous performance test among the intervention group (P<0.05).

Conclusion: Cognitive defects have negative impacts on the lives of of those surviving leukemia. Therefore, further studies are required to evaluate the role of cognitive rehabilitation in improvement of quality of life in this group of individuals.

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Background and purpose: Cancer is one of the leading causes of mortality worldwide. Acute promyelocytic leukemia, which is the most common type of blood cancer in adults, is the most prevalent type of acute leukemia. This type of cancer is characterized by uncontrolled proliferation, inhibited differentiation of immature myloied progenitors (promyelocytes), and accumulation of abnormal promyelocytes in the bone marrow. The aim of this study was to investigate the effects of Acroptilon repens L. extract on the growth rate of the acute promyelocytic leukemia cell line (HL-60).

Materials and methods: This is an experimental study. The HL-60 cells are round with large nuclei. These cells were maintained at 37°C in 5% CO₂ in RPMI-60 medium, supplemented with 10% fetal bovine serum, and 1% penicillin and streptomycin. The cytotoxic effects of Acroptilon repens extract on HL-60 cell line was investigated using microculture tetrazolium test (MTT). To identify the cytotoxic doses, different dilutions of the Acroptilon repens extract were added to 96-well culture plate, each containing 20 × 4 cells. The treated cells were incubated for 24, 48, and 72 h, and then the viability of the cells was assessed using MTT.

Results: According to the results, Acroptilon repens L. extract has cytotoxic effects in concentrations of 1, 2, 4, 6, and 8 mg/ml.

Conclusion: As the findings indicated, Acroptilon repens L. extract could be effective in growth rate of acute promyelocytic leukemia cell line (HL-60).

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Background and purpose: Interest into targeting PI3K in cancer has increased by the recent disclosure that aberrant activity of PI3K/Akt signaling pathway is associated with disease recurrence and poor outcome in different malignancies. The aim of this study was to investigate the potentiating effect of PI3K inhibitor, GS-1101 on doxorubicin-induced cell death in acute lymphoblastic leukemia-derived, Nalm-6 cells.
Materials and methods: In this experimental study, to evaluate whether abrogating PI3K/Akt pathway using GS-1101 could enhance cytotoxic effect of doxorubicin in acute lymphoblastic leukemia, Nalm-6 pre-B ALL cells were subjected to combination treatment and subsequent cell viability. Then cell count, metabolic activity, and transcriptional alteration of apoptosis-related target genes were investigated using Trypan blue exlussion, MTT and Rq-PCR analysis, respectively.
Results: Our data delineated that GS-1101 augments doxorubicin-induced cytotoxic and its anti-proliferative effect, as evidenced by the decreased cell survival, cellular metabolic activity, and reduction in the number of inhibitor-treated viable cells. Moreover, real-time PCR analysis revealed that induction of cell death by the drug combination was associated with increased Bax transcriptional activity (P≤0.01) followed by the elevated molecular ratio of Bax/Bcl-2 (P≤0.001).
Conclusion: This study suggested that abrogation of the PI3K pathway using GS-1101 could potentiate doxorubicin-induced anti-leukemic activity.
 

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Background and purpose: Telomerase activity has a major role in acute promyelocytic leukemia (APL). It also has a critical role in disease recurrence. This research aimed at studying the cytotoxic effects of telomerase inhibition using oligonucleotide-based molecule against human telomerase RNA template (hTERC antisense) and non-nucleoside small molecule targeting catalytic subunit (BIBR5132) on APL-derived cell line.
Materials and methods: To evaluate whether inhibition of telomerase using either hTERC antisense or BIBR5132 could exert cytotoxic effect in APL, NB4 cells were subjected to different concentrations of the inhibitors and subsequent cell viability, metabolic activity, induction of apoptosis were investigated using Trypan blue assay, MTT, and annexin/PI staining, respectively. Also, Caspase-3 enzymatic activity and transcriptional alteration of apoptosis-related target genes were investigated.
Results: We found that targeting telomerase using hTERC antisense (45 pmol/L) and BIBR1532 (75 µL) for 48 h reduced the survival rate of NB4 cells nearly by 30% and 40%, respectively and induced a caspase-3-dependent apoptosis. Our results also suggest that suppression of c-Myc and subsequent increment of Bax/Bcl-2 ratio coupled with decreased telomerase activity may be rational mechanisms for the cytotoxicity of both telomerase inhibitors against NB4 cells.
Conclusion: Current results clearly indicated that both BIBR1532 and hTERC antisense had anti-tumor activity against NB4 cells and anti-telomerase-based therapy may be an efficient treatment for acute promyelocytic leukemia.
 

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Background and purpose: Leukemia is a group of cancers caused by accumulation of malignant white blood cells in the blood or bone marrow. The aim of this study was to investigate the efficiency of hypnotherapy on pain relief, death anxiety, resilience, and healing of cancer cells in patients with acute myeloid leukemia treated with chemotherapy.
Materials and methods: A quasi-experimental study was carried out in which the research population were 86 patients of whom 26 (aged 30-50 years old) were selected via convenience sampling. They were randomly assigned into either experimental group or control group. Flow cytometry tests were done to confirm acute myeloid leukemia. The McGill Pain Management, Connor-Davidson Resilience scale, and the Collett-Lester Fear of Death Scale were administered to collect the data. Hypnosis therapy (six sessions) was done in experimental group. Data were analyzed applying analysis of covariance in SPSS V22.
Results: Hypnosis therapy was found to have significant effects on mental dimensions in experimental group compared to the control group (P= 0.039). Follow-up investigations showed more changes in death anxiety compared with other two dimensions in experimental group (6.67). Laboratory results indicated the onset of inflammatory reaction in experimental group.
Conclusion: Hypnotherapy is a powerful method in caring for cancer treatment.
 

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Background and purpose: Anti-apoptotic protein Bcl-2 prevents apoptosis through the mitochondrial pathway and high-level expression of Bcl-2 is required for survival of chronic lymphocytic leukemia (CLL). Berberine (BBR) is an alkaloid that induces apoptosis of cancer cells via mitochondrial pathways. The aim of this study was to investigate the effect of Berberine on Bcl-2 gene expression in peripheral blood mononuclear cells (PBMCs) of CLL patients compared to healthy subjects in vitro.
Materials and methods: In this experimental study, peripheral blood was obtained from 12 CLL patients and six age-matched healthy subjects. PBMCs were isolated by Ficoll separation and were cultured in two groups for 48 hours incubation; treated with Berberine (25 μM) and untreated. Finally, Bcl-2 gene expression was investigated in both groups following RNA extraction and cDNA synthesis by Real time PCR using SYBR Green method.
Results: Bcl-2 gene expression levels in CLL patients treated with Berberine were significantly lower than those of the untreated group (P<0.05), but no significant differences were found between healthy subjects treated with Berberine and control group.
Conclusion: The apoptotic activity of Berberine may be mediated through the reduction of Bcl-2 mRNA expression level in leukemic cells of CLL. Considering the major role of Bcl-2 in survival of tumor cells, Berberine could be considered as a natural therapeutic option for CLL.

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Background and purpose: Chronic lymphocytic leukemia (CLL) is the most common form of adult leukemia characterized by the accumulation of seemingly mature type B lymphocytes in peripheral blood and lymphatic organs. One of the main markers used in the diagnosis and prognosis of CLL is the CD38 gene. Polymorphism is considered to be a major genetic source of phenotypic change within a species and may be involved in developing the disease. Here, the study of CD38 gene polymorphism (rs 1800561) was done in terms of incidence and relevance to clinical and laboratory parameters in CLL patients.
Materials and methods: In order to confirm the CLL disease, CD5, CD19, and CD23 markers were investigated using immunophenotyping. CD38 gene polymorphism was studied in 70 patients with CLL and 70 healthy cases as control using a specific PCR method of polymorphism (Tetra-ARMS-PCR). Statistical analyses were performed in SPSS V18 applying Pearson Chi-Square test.
Results: In the patient group, we observed 97% wild-type genotype (CC), 1.5% heterozygote (CT), and 1.5 % homozygote (TT). The present study showed that 98.5% of the control subjects had wild-type (CC) and 1.5% had heterozygote (CT). The frequency of C and T alleles in both patients and control groups was 97.9%, 2.1%, 99.3%, and 0.7%, respectively.
Conclusion: Based on current study, despite differences in frequency of T and C alleles among patients and healthy individuals, no significant difference was found in the occurrence of this polymorphism and the risk of CLL.
 

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 This report describes an 89-year-old woman diagnosed with Philadelphia positive Chronic Myeloid Leukemia in 2007 who was initially treated with 200 mg/day imatinib. The patient demonstrated complete molecular response (CMR) in two tests in 2015 and 2018. During treatment between 2007 and 2019, despite increased dosage of imatinib and switching her therapy to nilotinib, complete hematological response was not obtained. ARMS-PCR was performed to find the possible cause of this failed response and JAK2 V617F mutation was detected. The patient with no BCR/ABL1 transcript showed a relatively good molecular response to nilotinib, although the complete hematological response was not observed. According to our findings, tyrosine kinase inhibitor could not be an effective therapy in this JAK2 V617F-positive MPN patient with a BCR/ABL1 mutation.

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Background and purpose: Melatonin is a well-known neuroendocrine reproductive hormone-based hormone that has an important role in the circadian clock system and has potent antioxidant properties. New studies have also suggested an antitumor role for melatonin. The role of melatonin receptors is proven in many cancers, including colon and breast cancers, but so far, no study has been performed in children with leukemia. To the best of our knowledge, this is the first study to investigate MT1A receptor polymorphism in children with leukemia using RELP-PCR.
Materials and methods: In this case-control study, 84 patients and 70 healthy individuals were included. After informed consent had been obtained, 10 ml of peripheral blood were taken from the participants. Genomic DNA was extracted using salting-out method and PCR-RFLP was used to evaluate the frequency of allelic and genotypic polymorphisms.
Results: The genotypes of MT1 gene rs2119882 polymorphism in control group included 40% TT genotype, 50% TC genotype, and 10% CC genotype. In patients, 40% had TT genotype, 35% had TC genotype, and 25% had CC genotype. Chi-square test showed a significant difference in genotypes between the groups studied (P<0.05).
Conclusion: This study confirmed the association between MT1 gene rs2119882 polymorphism and increased risk of childhood leukemia, Therefore, screening for melatonin receptor polymorphism can be helpful in prognosis and prevention of disease and appropriate treatment strategies in this cancer.

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 Background and purpose: Leukemia is one of the most common and deadly diseases which is an important health problem. The aim of this study was to evaluate the survival of patients with acute lymphoblastic leukemia (ALL) and acute myeloblastic leukemia (AML) using the penalized splines model and comparing that with the Cox proportional hazards model.
Materials and methods: This is a longitudinal survival research that used data from patients referred to Tehran Imam Khomeini Hospital between 2011 and 2014 and followed until 2016. Patients' information was collected through census, using a researcher-made checklist based on their files. Furthermore, the patients' latest status was obtained via phone calls. All medical records of the patients referred to Amir Al-Momenin hospital in Maragheh and diagnosed with prostate hyperplasia and underwent prostatectomy between 2014 and 2019 were examined taking a census. Kaplan-Meier nonparametric method was used to assess the 5-year survival rate, and Cox's semiparametric model was used to determine the risk factors in subgroups. Then, the penalized spline regression model was fitted using the R software version 4.2.2.
Results: The average age of patients was 36.29±16.01 and 36.7% of the patients were men. Half of the patients died, and 12.7% experienced recurrence. Acute lymphoblastic leukemia affected 34.7% of the patients, whereas acute myeloblastic leukemia affected 63.3%. The 5-year-survival rates for patients with ALL and AML were 18% and 29%, respectively. Age, white blood cell count, platelet count, and hemoglobin levels were found to be significantly associated with patients' survival (P<0.05).
Conclusion: Leukemia is seen at young ages in Iran, so, early detection and treatment of the disease can play a critical role in improving the survival of patients.
 

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Background and purpose: Chronic Lymphocytic Leukemia (CLL) is a lymphoproliferative disorder characterized by the clonal and malignant proliferation of mature B-cells within the peripheral blood, bone marrow, and secondary lymphoid tissues. In chronic infections and cancers, T-cells become exhausted due to continuous antigen exposure. This uncontrolled tumor cell growth creates an environment that lacks sufficient oxygen, nutrients, and physical space for normal cells, referred to as the tumor microenvironment. The tumor microenvironment employs multiple mechanisms to suppress the immune system, affecting metabolic pathways in both T-cells and tumor cells. Consequently, inhibitory receptors on T-cells and their ligands on tumor cells are upregulated, leading to reduced proliferation, cytotoxicity, and production of effector cytokines, such as interferon-gamma, by T-cells. T-cell exhaustion presents a therapeutic challenge in treating many malignancies, including CLL. In CLL, T-cell exhaustion is observed through the simultaneous upregulation of inhibitory receptors. Given that current therapies for CLL are often associated with side effects, resistance, and relapse, studying transcript changes may assist in designing more effective treatment strategies. This study investigates the transcriptome profile of CD8⁺ T-cells in CLL patients compared to healthy individuals using bioinformatics platforms.
Materials and methods: The keywords used in this study were CLL, CD8, and human. Raw data were initially searched in the SRA database using these keywords. Left and right reads were merged using CLC Genomics Workbench version 21. Following quality control of the data, sequences were assembled using human genome reference data, mRNA sequences, and coding sequences (CDS). Gene expression related to CD8+ T-cell exhaustion was then compared between CLL patients and healthy individuals. The analysis results were presented in heatmap and volcano plot formats.
Results: In this study, genes involved in CD8⁺ T-cell exhaustion and differential expression were identified by comparing 11 CLL patients with 11 healthy individuals. The expression levels of 59 selected genes were displayed in volcano plot and heatmap formats. Expression changes in 42 genes were statistically significant (FDR P<0.05), with 18 genes showing increased expression and 24 showing decreased expression in CLL patients compared to healthy controls.
Conclusion: The study found differences in the expression levels of genes involved in CD8⁺ T-cell exhaustion between CLL patients and healthy individuals. Molecules encoded by genes upregulated in CLL may serve as potential targets for developing new therapies. By creating monoclonal antibodies and small molecules to inhibit these targets, it may be possible to counteract T-cell exhaustion, representing a promising advance in the treatment of this phenomenon.