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Objective: To analyze and compare therapy responses, outcomes, and incidence of severe hematologic adverse events of flumatinib and imatinib in patients newly diagnosed with chronic phase chronic myeloid leukemia (CML) . Methods: Data of patients with chronic phase CML diagnosed between January 2006 and November 2022 from 76 centers, aged ≥18 years, and received initial flumatinib or imatinib therapy within 6 months after diagnosis in China were retrospectively interrogated. Propensity score matching (PSM) analysis was performed to reduce the bias of the initial TKI selection, and the therapy responses and outcomes of patients receiving initial flumatinib or imatinib therapy were compared. Results: A total of 4 833 adult patients with CML receiving initial imatinib (n=4 380) or flumatinib (n=453) therapy were included in the study. In the imatinib cohort, the median follow-up time was 54 [interquartile range (IQR), 31-85] months, and the 7-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.2%, 88.4%, 78.3%, and 63.0%, respectively. The 7-year FFS, PFS, and OS rates were 71.8%, 93.0%, and 96.9%, respectively. With the median follow-up of 18 (IQR, 13-25) months in the flumatinib cohort, the 2-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.4%, 86.5%, 58.4%, and 46.6%, respectively. The 2-year FFS, PFS, and OS rates were 80.1%, 95.0%, and 99.5%, respectively. The PSM analysis indicated that patients receiving initial flumatinib therapy had significantly higher cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) and higher probabilities of FFS than those receiving the initial imatinib therapy (all P<0.001), whereas the PFS (P=0.230) and OS (P=0.268) were comparable between the two cohorts. The incidence of severe hematologic adverse events (grade≥Ⅲ) was comparable in the two cohorts. Conclusion: Patients receiving initial flumatinib therapy had higher cumulative incidences of therapy responses and higher probability of FFS than those receiving initial imatinib therapy, whereas the incidence of severe hematologic adverse events was comparable between the two cohorts.
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Adulto , Humanos , Adolescente , Mesilato de Imatinib/efectos adversos , Incidencia , Antineoplásicos/efectos adversos , Estudios Retrospectivos , Pirimidinas/efectos adversos , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Resultado del Tratamiento , Benzamidas/efectos adversos , Leucemia Mieloide de Fase Crónica/tratamiento farmacológico , Aminopiridinas/uso terapéutico , Inhibidores de Proteínas Quinasas/uso terapéuticoRESUMEN
OBJECTIVE@#To explore the clinical significance of Epstein-Barr virus(EBV) detection and classification in peripheral blood of lymphoma patients.@*METHODS@#101 lymphoma patients were enrolled, the clinical characteristics of the patients were collected, including ages, sex, types of lymphoma, Ann Arbor stages, extranodal infiltration and lactate dehyhrogenase. Fluorescent quantitative PCR technology was used to detect the EBV-DNA. Polymerase chain reaction and Agarose gel electrophoresis was used for determination of EB genotyping. The difference between curative effect in EBV-DNA+ and EBV-DNA- patients, the correlation of adverse factors and EBV infection of the patients were analyzed.@*RESULTS@#68.3% (69/101) of the patients showed EBV-DNA positive. EBV-positive lymphoma patients showed more adverse prognostic factors than the patients with EBV-negative, which may lead to poorer disease outcome. Among the 46 B-cell non-Hodgkin's lymphoma patients, the overall response rate of EBV-positive patients (60.7%) was lower than EBV-negative patients(88.9%) (P<0.05); For 19 patients with Hodgkin's lymphoma, the overall response rate of EBV-positive patients (46.2%) was lower than EBV-negative patients (100%), the differences were statistically significant (P<0.05). Among 69 patients with EBV-infected lymphoma, 98.6% (68/69) showed type-2 EB virus, and 1.4% (1/69) were type-1 and type-2 mixed infections.@*CONCLUSION@#Most of EBV-positive in lymphoma patients were EBV type 2, patients with EBV-DNA+ shows poorer efficacy than EBV-DNA- patients.
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Humanos , ADN Viral , Infecciones por Virus de Epstein-Barr , Genotipo , Herpesvirus Humano 4/genética , Enfermedad de HodgkinRESUMEN
OBJECTIVE@#To evaluate the proformance of multiplex PCR and capillary electrophoresis(MPCE) in the detection of JAK2V617F and CALR mutation in myeloproliferative neoplasms(MPN).@*METHODS@#The specificity primers of JAK2617F gene mutation and the primers of CALR gene were designed at the same time. The JAK2V617F and CALR gene primers were labeled with Cy5 fluorescence, all the primers were mixed in one tube for multiplex PCR and the PCR prodcuts were analysised by capillary electrophoresis. Then detection limit and sensitivity of MPCE were evaluated, and compared with comercial diagnostic kit.@*RESULTS@#JAK2V617F and CALR gene mutations could be detect by MPCE in one PCR test. JAK2V617F mutation could be detected at 0.01 ng genomic DNA, double positive JAK2V617F and CLAR gene mutations could be detected at 0.1 ng genomic DNA, at least 0.1% JAK2V617F positive mutation could be detected. The consistency between MPCE and commercial diagnostic gene mutation kit was 100%.@*CONCLUSION@#It is developed that a new gene mutation detection method of JAK2 V617F and CLAR gene based on MPCE in our experiment and it can be used as a new reagent for molecular diagnosis of MPN patients.
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Humanos , Calreticulina/genética , Electroforesis Capilar , Janus Quinasa 2/genética , Mutación , Trastornos Mieloproliferativos/genética , Neoplasias , Pacientes , Reacción en Cadena de la PolimerasaRESUMEN
OBJECTIVE@#To investigate the chemotherapeutic efficency of quercetin sensitized adriamycin.@*METHOD@#CCK-8 was used to detect the inhibitory effect of different doses of adriamycin, quercetin and quercetin combined with adriamycin on the proliferation of primary leukemia cells from patients with clinically refractory acute leukemia. Quercetin, adriamycin and their combination were used to treat non-irradiated T-ALL leukemia mice to observe the changes of survival curve and myocardial injury.@*RESULT@#There was no significant difference in the inhibition rate of primary leukemia cell proliferation between the adriamycin concentration group (6, 0.6 and 0.06 μg/ml) and the adriamycin half-dose (3, 0.3 and 0.03 μg/ml) plus quercetin (0.25 mmol/L) group at three different time points (24, 48 and 72 hours). There was a significant difference in the inhibition rate of primary leukemia cell proliferation among the drug concentration groups, and the inhibition rate of primary leukemia cell proliferation was time-and concentration-dependent (r=0.995、r=1.000、r=0.984、r=0.993、r=0.999、r=0.960). In vivo experiments showed that the survival time of non-irradiated T-ALL leukemia mice treated with low-dose adriamycin combined with quercetin was not significantly prolonged compared with the high-dose adriamycin treatment group. The survival time of non-irradiated T-ALL leukemia mice treated with high dose of adriamycin and quercetin was significantly prolonged (P<0.05). Compared with adriamycin group, the SOD activity in adriamycin combined with quercetin group increased significantly and the MDA content decreased. The results of transcriptome sequencing analysis showed that the expression of Ighv1-84 and Igkv6-14 in adriamycin combined quercetin group and quercetin group was lower than that in adriamycin group. The Ms4a1, Podx1, Mecom, Sh3bgr12, Bex4 and Tdrp expression in adriamycin combined quercetin group and adriamycin group were higher than that in quercetin group, while Crabp1 expression was lower.@*CONCLUSION@#Quercetin can inhibit the proliferation of primary leukemia cells in a time-dependent manner. Quercetin combined with adriamycin inhibit the proliferation of primary leukemia cells significantly, and had synergistic and additive effects on the proliferation of primary leukemia cells, and the inhibiting effect of quercetin combined with adriamycin is concentration-and time-dependent. Quercetin combined with high-dose adriamycin can significantly prolong the survival time of non-irradiated T-ALL leukemia mice and reduce the myocardial damage caused by adriamycin.
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Animales , Humanos , Ratones , Apoptosis , Proliferación Celular , Doxorrubicina , Leucemia Mieloide Aguda , QuercetinaRESUMEN
OBJECTIVE@#To investigate the clinical significance of serum LRG1, LDH and β2-MG in the recurrence of diffuse large B-cell lymphoma(DLBCL) after chemotherapy.@*METHODS@#The serum levels of LRG1, LDH and β2-MG of 80 patients with DLBCL were detected before treatment and followed up for these patients was performed. The cut-off value of non-recurrent survival was determined by ROC analysis. The correlation of three serum markers for predicting recurrence with prognostic factors of diffuse large B-cell lymphoma patients after treatment was analyzed by ROC curve.@*RESULTS@#The serum levels of LDH, LRG1 and β2-MG were higher in the groups with high tumor stageing, extranodal invasion and bone marrow involvement, respectively(P<0.05). The optimal cut-off values for predicting recurrence risk determined by ROC analysis: LDH 402.37 U/L, LRG1 1.81 mg/L and β2-MG 168.3 ng/L, respectively.COX multivariate regression analysis showed that serum LRG1 was an independent factor affecting the recurrence of diffuse large B-cell lymphoma(P<0.05).@*CONCLUSION@#The serum level of LRG1 may become a new biological marker to predict the recurrence risk of diffuse large B-cell lymphoma.
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Humanos , Protocolos de Quimioterapia Combinada Antineoplásica , Glicoproteínas , Linfoma de Células B Grandes Difuso , Recurrencia Local de Neoplasia , Pronóstico , Estudios RetrospectivosRESUMEN
Objective To analyze the status of quality indicators(QI) on specimen acceptability and establish preliminary qual ity specification.Methods Web based External Quality Assessment system was used to collect data of laboratories partici pated in "Medical quality control indicators in clinical laboratory" from 2015 to 2017,including once in 2015 and 2017 and twice in 2016.Rate and sigma scales were used to evaluate incorrect sample type,incorrect sample container,incorrect fill level and anticoagulant sample clotted.The 25th percentile (P25) and 75th percentile (P75) of the distribution of each QI were employed to establish the high,medium and low specification.Results 5 346,7 593,5 950 and 6 874 laboratories sub mitted the survey results respectively.The P50 of biochemistry (except incorrect fill level),immunology and microbiology reach to 6σ.The P50 of clinical laboratory is 4 to 6σ except for incorrect sample container.There is no significant change of the continuous survey results.Based on results in 2017 to establish the quality specification,the P25 and P75 of the four QIs is 0 and 0.084 4 %,0 and 0.047 6 %,0 and 0.114 2 %,0 and 0.078 4 %,respectively.Conclusion According to the results of the survey,most laboratories had a faire performance in biochemistry,immunology and microbiology,and clinical laboratory needs to be strengthened.Laboratories should strengthen the laboratory information system construction to ensure the actual and reliable data collection,and make a long time monitoring to achieve a better quality.
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<p><b>OBJECTIVE</b>To investigate the influence of spleen on disease status of mouse T-ALL.</p><p><b>METHODS</b>The leukemia cells were transplanted into the mice, then the development levels of leukemia cells in different organs of transplanted mice were monitored at different time points after transplantation; the transplanted leukemia cell level in different organs was detected by flow cytometry at different time points after transplantation; the survival of transplanted mice was analyzed by means of splenectomy.</p><p><b>RESULTS</b>The spleen change displayed most severely in process of T-ALL, the number of T-ALL cells in the spleen obviously increased at initial period. The detection of organs showed that along with the progression of leukemia, spleen weight change was the most significant, following by the lever change. The splenectomy test showed that the spleen played a promotive role in progession of T-ALL, and the spleneetomy could difinitely postpone the progression of T-ALL in mice, there was significant difference between splenectomy and non-splenectomy.</p><p><b>CONCLUSIONS</b>In early stage after transplantation of T-ALL cells, the spleen has the promotive effect on function of T-ALL cells, which suggests that the spleen may be a important microenvironment for T-ALL cell migrating into body.</p>
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Animales , Ratones , Microambiente Celular , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Citometría de Flujo , Leucemia-Linfoma Linfoblástico de Células T Precursoras , Patología , Bazo , Patología , EsplenectomíaRESUMEN
This study was purposed to explore the anti-leukemic mechanism of quercetin (Que) in vivo and it enhancing chemotherapeutic effect of adriamycin (ADR) by establishing the quercetin-treated P388 transplanted nude mouse model. The P388 leukemic cells in logarithmic growth phase were taken and injected subcutaneously into BALB/c nude mice so as to establish the leukemia-transplanted nude mouse model. The model mice were treated by quercetin, ADR and their combination, and the survival changes of model mice were observed, the hemogram and peripheral blood cell count examination were performed regularly; the cell cycle was detected and the influence of quercetin on cell proliferation was analyzed by flow cytometry; the caspase-3 protein expression level was detected by ELISA; the mRNA and protein changes of NF-κB, BCL-2, BAX were measured by real-time quantitative flourascence PCR and Western blot respectively. The results indicated that the quercetin and adriamycin could significantly prolong the survival of P388 leukemia nude mice, and their combination displayed significantly prolonged effect. Quercetin and adriamycin alone or in their combination could reduce the ratio of G0/G1 phase in mice, the cell ratio in S phase and G2/M phase increased, and the effects of the combination group were more significant than that of the single agent groups. Quercetin could activate caspase-3 and promote leukemic cell apoptosis. Meanwhile, quercetin could down-regulate the expression of BCL-2 and NF-κB gene, and up-regulate the expression of BAX gene. It is concluded that through modulating the expression of apoptosis-related genes, the quercetin can inhibit leukemia cell proliferation, promote apoptosis, and enhance the chemotherapeutic effects of adriamycin. These results provide some valuable data for further research and development of quercetin as a new and effective anti-leukemic drug.
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Animales , Ratones , Apoptosis , Caspasa 3 , Ciclo Celular , Línea Celular Tumoral , Proliferación Celular , Doxorrubicina , Farmacología , Sinergismo Farmacológico , Leucemia , Quimioterapia , Patología , FN-kappa B , Quercetina , FarmacologíaRESUMEN
The study was aimed to investigate the effect of quercetin, flavonoid molecules on reversing leukemia multidrug resistance and its mechanism. K562/A cells were cultured in vitro with different concentrations of quercetin. Cell growth inhibition and adriamycin (ADR) sensitivity were detected by MTT method. Intracellular ADR concentration was determined by flow cytometry. Cell apoptosis was assayed by Annexin V/PI staining method. The expressions of drug transporter and apoptosis related genes were measured by real-time PCR array. The results indicated that quercetin inhibited the proliferation of K562 and K562/A in 5-160 µmol/L and with dose-dependent manner. Quercetin increased the sensitivity of K562/A cells to ADR in a low toxicity concentration. Flow cytometry showed that the quercetin increased the accumulation of ADR in K562/A cells when cells were co-cultured with 5 µmol/L ADR for 2 hours. Quercetin could induce the apoptosis of K562 and K562/A cells with dose dependent manner. Furthermore, some drug transport related genes such as ATP-binding cassette (ABC) and solute carrier (SLC) and some apoptosis-related genes such as BCL-2, tumor necrosis factor (TNF), tumor necrosis factor receptor (TNFR) families were down-regulated by quercetin. It is concluded that quercetin reverses MDR of leukemic cells by multiple mechanisms and the reversing effect is positively related to drug concentration.
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Humanos , Resistencia a Múltiples Medicamentos , Resistencia a Antineoplásicos , Células K562 , Quercetina , FarmacologíaRESUMEN
The purpose of this study was to explore the anti-leukemia effect of quercetin and kaempferol and its mechanism. The HL-60 cells were used as the leukemia models. The inhibitory effects of quercetin and kaempferol on growth of HL-60 cells was assessed by MTT assay. The effect of quercetin and kaempferol on cell cycle in HL-60 cells was detected by flow cytometry. The cytotoxic effect of these 2 drugs was analysed by single cell electrophoresis assay. Western blot analysis was used to study the apoptotic mechanism of HL-60 cells. The results showed that the quercetin and kaempferol had a significant anti-leukemia effect in vitro. The proliferation of HL-60 cells was significantly inhibited in dose-and time-dependent manners after treating with quercetin (r = 0.77) and kaempferol (r = 0.76) respectively, and the cytotoxicity of quercetin was superior to that of kaempferol. The quercetin and kaempferol induced G(2)/M arrest and apoptosis of HL-60 cells. The quercetin and kaempferol could down-regulate the survivin expression. It is concluded that the quercetin and kaempferol have significant anti-leukemia effect in vitro. Furthermore the apoptosis-inducing effect of quercetin is stronger than that of kaempferol, both of which induce apoptosis of HL-60 cells through depressing cell growth, arresting cell cycle and inhibiting expression of survivin.
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Humanos , Apoptosis , Ciclo Celular , Proliferación Celular , Regulación Leucémica de la Expresión Génica , Células HL-60 , Proteínas Inhibidoras de la Apoptosis , Metabolismo , Quempferoles , Farmacología , Quercetina , FarmacologíaRESUMEN
Objective To explore the development of eosinophilic inclusion bodies in PC12 cells incubated with proteasome inhibitor lactacystin. Methods PC12 cells were incubated for 24 h with 0, 5, 10 and 20 μmol/L lactacystin, and HE staining and α-synuclein immunohistochemistry was used to observe the changes of the inclusion bodies under light microscope. The changes in the inclusion bodies in the cytoplasm were also observed after treatment of PC12 cells with 10 μmol/l lactacystin for 24, 48, 72, 96 and 120 h. Results After incubation with lactacystin for 24 h, P12 cells showed increased number of eosinophilic inclusion bodies in the cytoplasm, and the increment appeared dose dependent. No inclusion bodies were observed in the cells without lactacystin treatment, and a few (3.33%±1.15%) occurred at lactacystin concentration of 5 μmol/L, numerous (71.33%±4.16%) at 10 μmol/l, and almost in every cell (90.33%±3.21%) at 20 μmol/L; some of the inclusion bodies were released from the cells. With the passage of time, inclusion bodies in the cells treated with 10 ?mol/l laetacystin began to dissociate with the cells, and the cytoplasm was reduced to contain only inclusion bodies and nucleus at 96 and 120 h. Immunohistochemistry showed that a -synuclein immunoreaetive granules concentrated and formed inclusion bodies close to the nucleus at 24 h. As the concentration increased and time prolonged, α-synuclein-immunoreactive inclusion bodies were released from the cells, leaving only inclusion bodies and cell nuclei presented at 96 and 120 h. Conclusion Incubation with laetacystin can induce cytoplasmic eosinophilic and α-synuclein-immunoreaetive inclusion bodies in PC12 cells. The development of inclusion bodies in this model is consistent with that in human Parkinson's disease and diffused Lewy body disease. This model is helpful for further study of the mechanism of neurodegenerative diseases.
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This preject is to explore the reversal efficacy of calmodulin antagonist berbamine (BBM) on multidrug resistance (MDR) and its mechanism. Human erythroleukemic cell line K562 and its adriamycin-resistant counterpart K562/A02 were used in the study. The cells were co-cultured with ADR and BBM in different concentrations. MTT assay was used to analyze the effect of BBM on cell growth inhibition. According to the MTT assay, the 50% inhibitory concentration (IC(50)), the multiples of drug resistance and increased sensitivity of ADR were calculated. The concentration of intracellular ADR and expression level of P-gp were detected by flow cytometry (FCM). The expression level of mdr1 mRNA and survivin mRNA was detected by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) with beta-actin as internal reference. The results showed that IC(50) of ADR in K562 and K562/A02 cells was 1.16 +/- 0.09 micro mol/L and 37.47 +/- 1.76 micro mol/L, respectively. The resistant multiple of K562/A02 cells to ADR was 32.30 higher than that of K562 cells. BBM increased the chemo-sensitivity of ADR in K562/A02 cells with dose-dependent relationship, i.e. when 5, 10 and 20 micro mol/L BBM was added in the culture the chemo-sensitivity of ADR was increased to 2.01-, 9.68-, and 41.18-fold (P < 0.01), respectively. After treating K562/A02 cells by 5 or 10 micro mol/L BBM for 2 hours the accumulation of intracellular ADR was increased to 1.41- and 1.52-fold (P < 0.01), respectively. Treating by BBM for 72 hours decreased 4.12% (P < 0.05) and 27.09% (P < 0.01) of P-gp expression, respectively, meanwhile down-regulated expression of mdr1 mRNA and survivin mRNA was found. In conclusion, BBM could increase intracellular concentration of ADR in K562/A02 that down-regulated expression level of mdr1 mRNA and P-gp and survivin so that the sensitivity of K562/A02 to ADR was increased significantly.