ABSTRACT
Chimeric antigen receptor T (CAR-T) cell therapy is a novel cellular immunotherapy that is widely used to treat hematological malignancies, including acute leukemia, lymphoma, and multiple myeloma. Despite its remarkable clinical effects, this therapy has side effects that cannot be underestimated. Cytokine release syndrome (CRS) is one of the most clinically important and potentially life-threatening toxicities. This syndrome is a systemic immune storm that involves the mass cytokines releasing by activated immune cells. This phenomenon causes multisystem damages and sometimes even death. In this study, we reported the management of a patient with recurrent and refractory multiple myeloma and three patients with acute lymphocytic leukemia who suffered CRS during CAR-T treatment. The early application of tocilizumab, an anti-IL-6 receptor antibody, according to toxicity grading and clinical manifestation is recommended especially for patients who suffer continuous hyperpyrexia, hypotensive shock, acute respiratory failure, and whose CRS toxicities deteriorated rapidly. Moreover, low doses of dexamethasone (5-10 mg/day) were used for refractory CRS not responding to tocilizumab. The effective management of the toxicities associated with CRS will bring additional survival opportunities and improve the quality of life for patients with cancer.
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Objective To screen the optimal RNA interference sequence of acute monocytic leukemia associated antigen 22 (MLAA-22) gene in order to study gene function of it. Methods MLAA-22 coding sequence (CDS) was cloned by reverse transcription polymerase chain reaction (RT-PCR) and the CDS was inserted in to pEGFP-N1-3FLAG vector to construct eukaryotic expression vector of MLAA-22. At the same time, four RNA interference sequences were designed and cloned to the vector. Expression vector and RNA interference vector were co-transfected into 293T cells, and the optimal RNA interference sequence was screened by fluorescence and Western blot analyses. Results MLAA-22 eukaryotic expression vector pEGFP-N1-3FLAG and four RNA interference vectors were successfully constructed. After co-transfected 293T cells, KD2 was selected as the optimal interference sequence of MLAA-22. Conclusion KD2 is an optimal interference sequence for targeting MLAA-22 antigen gene.
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Objective To compare the differences of insomnia prescription laws between ancient and mordern acupuncture. Methods Collected ancient and mordern acupuncture treatment of insomnia prescriptions literature, using fourteen frequency, percentage of acupuncture acupoints, the points of the distribution and distribution in different parts of the body, the specific acupoint application and prescription features for statistical description. Results The ancient physicians selected points more dispersed, acupoints are mainly concentrated in the chest , abdomen and leg. Modern physicians point selection is relatively concentrated, prescription relatively fixed, the acupoints are mainly concentrated in the head and neck and lower limb;acupoints are mainly concentrated in the Urinary bladder meridian of foot Taiyang, the ancient physicians also more choice of Ren pointss, modern physicians are more optional Du points. Conclusion The physicians prescriptions are quite different, but in the meridian points and the type of use is similar, mostly concentrated in the selection of points Urinary bladder meridian of foot Taiyang, especially the five Shu points. Du, Ren, Spleen meridian and Stomach meridian of Foot Yangming using positive significance for the treatment of insomnia by acupuncture choice.
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Objective To investigate the expression level and clinical significance of WT1 gene in acute luekemia (AL) patients.Methods WT1 gene level was detected by real time quantitative-polymerase chain reaction in acute myelogenous leukemia (AML) and in acute lymphocytic leukemia (ALL) patients.Then the expression levels of WT1 gene in different subtypes of AML were compared,and the correlation between gene expression and disease courses and prognosis were observed.Moreover,the relationship between disease courses and WT1 expression in patiens after receiving haemopoietic stem cell transplantation were analyzed.Results Among 66 cases,WT1 expression positive rate was 87.5 ％ (14/16) in AML and 76.0 ％ (38/50) in ALL.In AML,the expression level in M3 showed the lowest than that in any other subtypes (compared with M1,M2,M4,M5,P value was 0.040,0.007,0.006 and 0.01,respectively).The expression level of WT1 was closely correlated with leukemia disease courses.The expression level in complete remission (CR) group showed a significant lower expression level than that in non-remission group (P =0.018) and relapse group (P =0.003),and the re-increase of WT1 expression level could predict relapse as early as 1.5 months.Moreover,WT1 expression also showed an close relationship with prognosis of patients receiving haemopoietic stem cell transplantation.Patients whose WT1 was undetectable had a better prognosis than those with persistent expression,and increase again after becoming undetectable.Conclusion WT1 has a high expression level in AL,which can represent minimal residual disease.The expression level in M3 was lowest than that in different AML subtypes,and its expression level has a close correlation with clinical disease course and prognosis of AL.
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[Objective]To discuss the clinical effect of fludarabine and busulfan (Bu+Flu) as a low toxicity myeloablative conditioning regimen for allogeneic peripheral blood stem cell transplantation (allo-HSCT)in leukemia patients.[Methods]Clinical data of 13 patients with hematological malignancies receiving conditioning regimen with Bu+Flu for allo-HSCT were analyzed retrospectively.Conditioning regimen was Bu+Flu,compalriot mismatched and unrelated transplantation combined with rabbit anti-human thymocytes immune globulin (ATG).CsA+short course of methotrexate or CsA + mycophenolate mofetil were used to prevent graft-versus-host disease (GVHD).DNA sequencing of short tandem repeat (STR)polymorphism analysis method was performed for identification of donor stem cells implantation.[Results]13 patients all tolerated with this conditioning regimen well,no serious complications occurred.Neutrophil engraftment was at 9-15 days (median 11 days),platelet engraftment at 8-25 days (median 13 days).10 patients achieved hematopoiesis reconstitution with their full donor chimerisms confirmed by STR-DNA analysis.Acute GVHD occurred in 5 cases,accounting for 38.5％.Chronic GVHD occurred in 4 cases of 10 patients could be assessed,accounting for 40.0％.Severe GVHD more than Ⅱ degree did not happen.1-39 months (median time 11 months)of follow-up revealed the overall survival rate of 76.9％(10/13),disease-free survival of 61.5％ (8/13).The causes for death were relapse in all.[Conclusion]The conditioning regimen with Bu+Flu has low toxicity,well tolerance and better effect.
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Objective To study the clinic effect and safety of MEAD chemotherapy regimen for adult patients with relapsed or refractory acute lymphocyte leukemia. Methods Between July 2006 and July 2009,twenty-two adult patients with relapsed or refractory acute lymphocyte leukemia received MEAD regimen (mitoxantrone 6 mg/d dl-3 iv drip,cytarabine 100 mg/d dl-5 iv drip,etoposide 100 mg/d dl-5 iv drip,dexmethasone 10 mg/d dl-8 iv drip). Results The complete remission (CR) rate of adult patients with relapsed or refractory acute lymphocyte leukemia was 31.8 %,the partial remission(PR) rate was 22.7 % and the overall response (OR) rate 54.5 %. The cumulitive CR rate was 50.0 %,and the PR rate 40.9 % after two times MEAD chemotherapy regimen. The main adverse effect was different level of myelosuppression,and other toxicity of vital organ was mild. Conclusion MEAD regimen is effective and can be tolerated for adult patients with relapsed or refractory acute lymphocyte leukemia,and its side effect is mild.
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Objective To study the curative effects and adverse effects of the thalidomide combined with COMP chemotherapy in treating multiple myeloma(MM).Methods 42 patients were initially diagnosed as MM and 27 patients were refractory and relapsed MM.The small dose of thalidomide combined with COMP management and COMP management alone were used.The effective rate and adverse effects were analyzed.Changes of M-protein in serum,percentage of plasma cells in bone marrow and the level of hemoglobin were also analyzed in both pre-treatment and post-treatment periods.Results In 42 patients who were initially diagnosed as MM,the effective rate was 40.9% for 22 patients treated by chemotherapy alone and 70.0% for 20 patients treated by the thalidomide combined with chemotherapy.Statistic difference was observed between those two group.As to the 27 patients who were refractory and relapsed MM,the effective rate was 42.9% for 13 patients treated by chemotherapy alone and 84.6% for 14 patients treated by the thalidomide combined with chemotherapy.Statistic significance was present between them.Adverse effects were less and tolerated.Conclusion Treatment of small dose of thalidomide combined with COMP chemotherapy could significantly improve the effective rate with less adverse effects.
ABSTRACT
Human acute premyeloid leukemia cell cDNA expression library was constructed to screen acute premyeloid leukemia tumor antigen. Total RNA and purified mRNA were extracted from human premyeloid cell line NB4. First and second strands of cDNA were synthesized by reverse transcription. After blunting, the cDNA fragments were ligated with EcoR I adapters. Then the cDNAs were digested with Xho I, and less than 400 bp cDNA fragment was removed by Sephacryl-S400 spin column, the remaining were ligated with lambdaZAP vector. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli XL1-Blue-MRF' for titration. The recombinants were examined by color selection. In order to evaluate the size of cDNA inserts and the diversity of library, the pBK-CMV phagemid was excised from the ZAP express vector by using ExAssist helper phage with XLOLR strain, and then the pBK-CMV phagemid was digested by Xho I and EcoR I. The results showed that the NB4 cell line cDNA library consisting of 1.65 x 10(6) recombinant bacteriophages was constructed with the recombinant ratio of 99.6%. The average length of the recombinant exogenous inserts was about 1.7 kb. It was concluded that the constructed cDNA library are deserved to screen target clones.
Subject(s)
Bacteriophages/genetics , DNA, Complementary/biosynthesis , DNA, Neoplasm/biosynthesis , DNA, Recombinant/biosynthesis , Gene Library , Genetic Vectors , Leukemia, Promyelocytic, Acute/genetics , Leukemia, Promyelocytic, Acute/metabolism , Leukemia, Promyelocytic, Acute/pathology , RNA-Directed DNA Polymerase/metabolism , Transcription, Genetic/geneticsABSTRACT
Human acute premyeloid leukemia cell cDNA expression library was constructed to screen acute premyeloid leukemia tumor antigen. Total RNA and purified mRNA were extracted from human premyeloid cell line NB4. First and second strands of cDNA were synthesized by reverse transcription. After blunting, the cDNA fragments were ligated with EcoR I adapters. Then the cDNAs were digested with Xho I, and less than 400 bp cDNA fragment was removed by Sephacryl-S400 spin column, the remaining were ligated with lambdaZAP vector. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli XL1-Blue-MRF' for titration. The recombinants were examined by color selection. In order to evaluate the size of cDNA inserts and the diversity of library, the pBK-CMV phagemid was excised from the ZAP express vector by using ExAssist helper phage with XLOLR strain, and then the pBK-CMV phagemid was digested by Xho I and EcoR I. The results showed that the NB4 cell line cDNA library consisting of 1.65 x 10(6) recombinant bacteriophages was constructed with the recombinant ratio of 99.6%. The average length of the recombinant exogenous inserts was about 1.7 kb. It was concluded that the constructed cDNA library are deserved to screen target clones.