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Objective:To investigate the influence of dual-task treadmill training on the motor function of children with spastic hemiplegic cerebral palsy(CP).Methods:A prospective study was carried out on 36 children with spastic hemiplegic CP admitted to the Department of Rehabilitation, Qingdao Women and Children′s Hospital from March 2020 to August 2021.The subjects were divided into the control group (18 cases) and the experimental group (18 cases) by the random number sequence method.During the intervention, 2 cases in the control group dropped out of the study due to cough and fever.Finally, there were 16 cases left in the control group and 18 cases in the experimental group.Both groups received the same routine rehabilitation training.Additionally, the control group were given single-task treadmill training, while the experimental group were treated dual-task treadmill training.Before training and after 4 weeks of training, the children in the two groups were evaluated and compared by using Gross Motor Function Measure-88 (GMFM-88), Pediatric Balance Scale (PBS), Maximum Walking Speed test(MWST), single-task modified Timed Up and Go test (mTUG) and dual-task mTUG.Gender and Gross Motor Function Classification System(GMFCS) classification in general data were compared by using the Chi-square test or Fisher′ s exact test.Age and indices of rehabilitation assessment were compared by using the paired-samples t-test within groups and indepen-dent-samples t-test between the two groups. Results:There was no significant difference in the evaluation data of the GMFM-88 D score [(34.25±1.61) points vs.(34.56±1.76) points], GMFM-88 E score [(50.53±7.20) points vs. (50.61±6.75) points], PBS score [(39.06±4.34) points vs. (38.89±4.44) points], MWST time [(12.69±3.07) s vs. (13.56±2.97) s], single-task mTUG time [(11.38±2.58) s vs. (11.94±2.51) s], and dual-task mTUG time [(30.06±8.08) s vs. (31.50±8.56) s]between the control group and the experimental group before training (all P>0.05). After 4 weeks of training, the GMFM-88 score of the control group was (35.88±1.82) points in the D dimension and (51.20±6.64) points in the E dimension.Besides, the PBS score of the control group was (40.75±4.14) points, the MWST time was (10.81±2.95) s, and the single-task mTUG time was (10.06±2.52) s. As for the experimental group, the GMFM-88 score was (36.28±1.99) points in the D dimension and (53.94±6.98) points in the E dimension, the PBS score was (43.06±4.94) points, the MWST time was (10.44±2.83) s, and the single-task mTUG time was (10.56±2.73) s. The evaluation indexes of the two groups after training for 4 weeks were significantly better than those before training ( t=-3.058, -2.197, -7.132, 1.235, 2.952 in the control group, and t=-5.953, -12.432, -8.333, 3.149, 7.578 in the experimental group, all P<0.05). There was no significant difference in GMFM-88 scores in D and E dimensions, PBS scores, MWST time and single-task mTUG time between the two groups ( P>0.05). The dual-task mTUG time of the control group was (29.10±8.28) s after 4 weeks of training, which was not statistically different from that before training ( t=1.578, P>0.05). The dual-task mTUG time of the experimental group was (23.06±7.30) s after 4 weeks of training, which was significantly better than that before training ( t=13.930, P<0.05) and that of the control group ( t=2.296, P<0.05). Conclusions:Both single-task and dual-task treadmill training can remarkably improve the motor function of children with spastic hemiplegic CP.Single-task training cannot improve the motor function of children with spastic hemiplegic CP in the dual-task condition, while dual-task training can effectively improve the motor function with spastic hemiplegic CP in the dual-task condition.
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Objective:To explore the efficacy and safety of recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) and human granulocyte colony-stimulating factor (G-CSF) for the prevention of post-chemotherapy infections in pediatric hematologic neoplasms.Methods:A total of 134 children hospitalized for chemotherapy in 6 tertiary hospitals from July 2016 to June 2018 were collected, including 60 cases in Children's Hospital of Fudan University, 38 cases in Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, 29 cases in Children's Hospital Affiliated to Soochow University, 4 cases in the Affiliated Hospital of Qingdao University, 2 cases in Northwestern Women and Children's Hospital, and 1 case in Shandong Provincial Qianfoshan Hospital. The children were divided into GM-CSF group (38 cases), G-CSF group (45 cases) and GM-CSF+G-CSF group (51 cases) by using random number table method. The incidence of infections, the recovery time of absolute neutrophil counting (ANC), the decrease of blood platelet count (Plt) and the incidence of adverse reactions were compared among the three groups.Results:In all children, a total of 64 cases (47.8%) had infections during the myelosuppression phase after chemotherapy, of which 18 cases (47.4%) in GM-CSF group, 20 cases (44.4%) in G-CSF group, and 26 cases (51.0%) in GM-CSF+G-CSF group. The incidence of respiratory infection in G-CSF group was higher than that in GM-CSF group and GM-CSF+ G-CSF group [22.2% (10/45) vs. 2.6% (1/38), 4.0% (2/51), χ2 = 12.00, P = 0.002]. The median time to recovery of ANC > 1.5×10 9/L was 10.5 d (8 d, 15 d) in all children, 12 d (10 d, 16 d) in GM-CSF group, 9 d (8 d, 12 d) in G-CSF group, and 10 d (8 d, 16 d) in GM-CSF+G-CSF group. In all children, a total of 101 cases (75.4%) had Plt<50×10 9/L during the myelosuppression phase, and 79 cases (59.0%) had Plt <20×10 9/L. The differences in the incidence of Plt <50×10 9/L and <20×10 9/L among the three groups were not statistically significant (both P > 0.05). In all children, the adverse reactions occurred in 24 cases (17.9%), including 20 cases (14.9%) of fever, 2 cases (1.5%) of sore throat, 1 case (0.7%) of nausea, and 1 case (0.7%) of diarrhea; no adverse reactions of grade 2 or above occurred. The difference in the incidence of adverse reactions among the three groups was not statistically significant ( P>0.05). Conclusions:The efficacy of GM-CSF and G-CSF for the prevention of infections in pediatric hematologic neoplasms during the myelosuppression phase after chemotherapy is roughly equivalent, and combination of both has a good tolerance. The incidence of respiratory infection using GM-CSF alone or GM-CSF+G-CSF is low, which might benefit from the effect of GM-CSF on lung infections.
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Objectives:To analyze the prevalence and risk factors of chronic nephropathy among hospitalized gout patients.Methods:537 hospitalized gout patients in Tianjin Medical University Chu Hsien-I Memorial Hospital were enrolled. According to the levels of glomerular filtration rate (eGFR>90 ml/min, 60-90 ml/min and <60 ml/min), patients were divided into normal group, mild chronic nephropathy group and moderate-severe chronic nephropathy group. The height, weight, waist, hip and blood pressure of the patients were measured. Blood samples from elbow vein were collected to measure biochemical indexes. The 24-hour urine was collected to measure creatinine and uric acid. Multivariate regression analysis was used to analyze the related factors that may affect eGFR.Results:The prevalence of chronic nephropathy among gout patients was 46.76%(251/537), which the prevalence of mild chronic nephropathy was 34.82%(187/537), and moderate severe chronic nephropathy was 11.92%(64/537). With the decline of eGFR, the older the patients, the longer the course of gout and hypertension, the more the number of joints involved, the higher the blood uric acid, urinary microalbumin, urinary protein, white blood cell count, uric acid excretion fraction (FeUA), and the lower the red blood cell count, hemoglobin and serum albumin. Pearson correlation analysis of eGFR showed that age, duration of gout and hypertension, number of involved joints, hemoglobin, serum albumin, serum uric acid, urinary microalbumin, and FeUA were correlated with eGFR ( r=-0.43, -0.238, -0.25, -0.128, 0.155, 0.138, -0.169, -0.29, -0.372, respectively, P<0.01). Multivariate regression analysis showed that age, the course of gout, serum uric acid, microalbuminuria and FeUA were independent risk factors for eGFR decline ( P<0.01). Conclusions:Eldly, long course of gout, poor control of serum uric acid and increased excretion of uric acid and microalbumin are risk factors of gout.
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Four hundred and four male patients with primary gout were enrolled. According to the degree of nonalcoholic fatty liver diseases (NAFLD), the patients were divided into simple gout ( n=121), gout combined with mild NAFLD ( n=149) and gout combined with moderate-severe NAFLD ( n=134). The height, weight, waist, hip, blood pressure and blood biochemistry parameters of patients were measured. The degree of NAFLD was negatively correlated with the age of patients in three groups. The BMI, ratio of waist/hip, count of red cells, hemoglobin, hematocrit, red blood cell distribution width ( SD and CV), triglyceride, alanine aminotransferase and HOMA-IR were increased with the increasing of NAFLD severity (all P<0.05). Red blood cell count, hemoglobin, alanine aminotransferase, serum uric acid increased with the increasing of NAFLD severity (all P<0.05). Platelet, serum urea nitrogen and serum creatinine were decreased with the increase of NAFLD severity. Logistic regression showed that BMI, hemoglobin and HOMA-IR were independent risk factors for NAFLD. The prevalence and the severity of NAFLD was increased with increasing quadrates of hemoglobin. Taking group Q1 as a control, OR of NAFLD in group Q2 was 1.166(95 %CI:0.638-2.133), OR in group Q3 was 2.011(95 %CI:1.122-3.605)and OR in group Q4 was 3.120(95 %CI:1.613-6.034). The result indicates that hemoglobin levels are associated with the development and the severity of NAFLD in male patients with primary gout.
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Hemophilic arthropathy (HA) is one of the most common and typical manifestation in the course of recurrent bleeding episodes in patients with hemophilia.The main pathogenesis are synovial hyperplasia, inflammation and neovascularization, articular cartilage degeneration and bone injury.Considering its pathogenesis, potential targets for disease modifying therapy in HA are iron, inflammation, vascular remodeling, hyper fibrinolysis, bone remodeling and cartilage regeneration.So far, iron chelators, anti-inflammatory therapy, anti-fibrinolytics and bone remodeling agents have demonstrated beneficial effects, predominantly in a preclinical setting.This article reviews the pathogenesis of HA and identifies potential therapeutic targets to provide more therapeutic options for HA.
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Hemophilic arthropathy (HA) is one of the most common and typical manifestation in the course of recurrent bleeding episodes in patients with hemophilia.The main pathogenesis are synovial hyperplasia,inflammation and neovascularization,articular cartilage degeneration and bone injury.Considering its pathogenesis,potential targets for disease modifying therapy in HA are iron,inflammation,vascular remodeling,hyper fibrinolysis,bone remodeling and cartilage regeneration.So far,iron chelators,anti-inflammatory therapy,anti-fibrinolytics and bone remodeling agents have demonstrated beneficial effects,predominantly in a preclinical setting.This article reviews the pathogenesis of HA and identifies potential therapeutic targets to provide more therapeutic options for HA.
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Objective@#To evaluate the long-term efficacy and prognostic factors of childhood acute lymphoblastic leukemia (ALL) enrolled in Shanghai Children's Medical Center-Acute Lymphoblastic Leukemia-2005(SCMC-ALL-2005) multicenter study.@*Methods@#Between May 2005 and December 2014, 1 497 newly diagnosed ALL patients were enrolled and treated in 5 hospitals of SCMC-ALL-2005 study group, using risk-stratified SCMC-ALL-2005 protocol. Risk group classification and treatment intensity were based on clinical features, genetic abnormalities, early response to treatment and levels of minimal residual disease (MRD). Kaplan-Meier method was used to generate overall survival (OS) and event-free survival(EFS) curves. Cox proportional hazards models were used for multivariate analyses.@*Results@#The patients were followed up to December 31, 2016, the median follow-up time was 69 months (24-141 months). The 5-year and 10-year OS rates were (80.0±1.0)% and (76.0±2.0)%. The 5-year and 10-year EFS rates were (69.0±1.0)% and (66.0±2.0)%. The 5-year and 10-year relapse rates were (23.0±1.0)% and (25.0±2.0)%. The 5-year OS and EFS for low risk (LR), intermediate risk (IR) and high risk (HR) were (91.1±1.4)% and (83.3±1.8)%, (79.2±1.5)% and (68.9±1.7)%, (52.9±4.4)% and (30.0±3.8)%, respectively. MRD negative status (<0.01%) on day 55 was seen in 792 patients (82.8%) and positive MRD on day 55 was associated with poor prognosis (OR=1.9, 95%CI: 1.3-2.7, P=0.001). Twenty-four HR patients received allogeneic hematopoietic stem cell transplantation and 17(70.8%) of them were alive and in remission. A total of 164 severe adverse events occurred, 46 of them died, treatment-related mortality was 3.1%.@*Conclusions@#In this large sample research, the overall outcome for multi-center SCMC-ALL-2005 study was favorable. This helps to promote the standardized treatment of childhood ALL to the whole country. MRD results on day 55 of induction therapy have important prognostic and therapeutic implications.
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Objective To study the effect of a new immunomodulator composed of muramyl dipeptide(MDP) and anti-CD10monoclonal antibody(MDP-Ab)on the dendritic cells(DC)of children with acute leukemia. Methods DC was adopted to divide the children with acute lymphoblastic leukemia into 6 groups,including the control group,unconjugated anti-CD10alone,unconjugated MDP alone,MDP-Ab alone,lipopolysaccharide(LPS)alone and MDP-Ab + LPS.The immunophenotypes,the endocytosis interleukin-12(IL-12)were detected.The stimulation index of autologous lymphocytes was assayed by adopting 5-(and 6)-carboxyfluorescein diacetate,succinimidyl es-ter(CFSE)-staining method.The supernatants of DC and autologous lymphocytes were used to detect the level of in-terferon-γ(IFN-γ)by using enzyme-linked immunosorbent assay.Results (1)DC immunophenotype:The ex-pressions of human leukocyte antigen-DR(HLA-DR),mature molecule(CD83)and co-stimulatory molecules (CD80and CD86)were increased significantly upon DC triggered with MDP-Ab,compared with the control group,un-conjugated anti-CD10group,and unconjugated MDP group,but lower than those in LPS and combination of MDP-Ab with LPS(F=629.62,P=0.000).(2)The level of IL-12:a significant increase in IL-12 level was detected in MDP-Ab group,LPS group,and combination of MDP-Ab with LPS group,compared with the control group,uncon-jugated anti-CD10group,and unconjugated MDP group(F=857.87,P=0.000). There were significant differences among the first three groups.(3)Endocytosis assay:The uptake of DCs stimulated by unconjugated anti-CD10,un-conjugated MDP,MDP-Ab immunoconjugate,LPS or combination was lower than that of immature DC in the control group which was(81.3 ± 10.1)%.(4)Mixed lymphocyte reaction and IFN-γ level:DC,treated with MDP-Ab, LPS and combination,stimulated more CFSE positive cells and higher level of IFN-γ secretion than the control group and unconjugated anti-CD10group,unconjugated MDP group. The most significance was observed in combination of MDP-Ab with LPS(F=393.36,P=0.000;F=2 497.18,P=0.000).Conclusion It is concluded that MDP-Ab could promote the proliferation and maturation of DC derived from blood of children with acute leukemia.
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Objective To compare the efficacy and safety of humidified high-flow nasal cannula (HHFNC)and nasal continuous positive airway pressure (nCPAP) for moderate and severe bronchiolitis treatment in infants.Methods Thirty-two infants who were diagnosed as moderate and severe bronchiolitis admitted to Department of Pediatric Critical Care Medicine,the Affiliated Hospital of Qingdao University from September 2016 to May 2017 were selected,and they were randomly assigned into HHFNC group(17 cases) and nCPAP group(15 cases).The heart rate,breathing,oxygen saturation (SpO2),arterial partial pressure of oxygen [Pa (O2)],partial pressure of carbon dioxide [Pa (CO2)],pH value after treatment for 12 h,the duration of non-invasive ventilation and the incidence rates of invasive ventilation use,complications were compared between the 2 groups.Results After the treatment for 12 h,in HHFNC group,the heart rate was (130.88 ± 2.87) times/min,respiratory rate was (37.35 ± 3.55) times/min,SpO2 was(97.06 ± 1.43)％,pa(O2) was (99.65-±8.07) mmHg,pa(CO2) was (35.88 ±4.27) mmHg,pH was 7.42 ± 0.03;while in the nCPAP group,the heart rate was (135.73 ± 6.29) times/min,respiratory rate was (41.40 ± 4.40)times/min,SpO2 was (96.00 ± 1.13) ％,Pa (O2) was (91.33 ± 9.45) mmHg,pa (CO2) was (40.13 ± 3.72)mmHg,pH was 7.39 ± 0.03.The breathing,heart rate,oxygen saturation and arterial blood gas in both groups after treatment were improved significantly compared with those before treament,and the differences were statistically significant (all P ＜ 0.05);after treatment for 12 h,the heart rate,breathing,oxygen saturation and arterial blood gas of HHFNC group were improved more than those of the nCPAP group,and the differences had statistical significance (all P ＜ 0.05).The duration of non-invasive ventilation was (45.88-± 6.49) hours in HHFNC group,and (49.33 ± 8.99) hours in nCPAP group,so there was no difference between the 2 groups (t =1.254,P =0.219).There was 12 cases (80.0％) of mild complication in nCPAP group and 5 cases(29.4％) in HHFNC group,while the incidence rate of invasive ventilation use was 3 cases(20.0％) in nCPAP group and 1 case (5.9％) in HHFNC group.Conclusion The efficacy and security of HHFNC on moderate and severe bronchiolitis are better than those of nCPAP,and it is recommended for clinical application widely.
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Objective To compare the efficacy and safety of humidified high-flow nasal cannula (HHFNC)and nasal continuous positive airway pressure (nCPAP) for moderate and severe bronchiolitis treatment in infants.Methods Thirty-two infants who were diagnosed as moderate and severe bronchiolitis admitted to Department of Pediatric Critical Care Medicine,the Affiliated Hospital of Qingdao University from September 2016 to May 2017 were selected,and they were randomly assigned into HHFNC group(17 cases) and nCPAP group(15 cases).The heart rate,breathing,oxygen saturation (SpO2),arterial partial pressure of oxygen [Pa (O2)],partial pressure of carbon dioxide [Pa (CO2)],pH value after treatment for 12 h,the duration of non-invasive ventilation and the incidence rates of invasive ventilation use,complications were compared between the 2 groups.Results After the treatment for 12 h,in HHFNC group,the heart rate was (130.88 ± 2.87) times/min,respiratory rate was (37.35 ± 3.55) times/min,SpO2 was(97.06 ± 1.43)％,pa(O2) was (99.65-±8.07) mmHg,pa(CO2) was (35.88 ±4.27) mmHg,pH was 7.42 ± 0.03;while in the nCPAP group,the heart rate was (135.73 ± 6.29) times/min,respiratory rate was (41.40 ± 4.40)times/min,SpO2 was (96.00 ± 1.13) ％,Pa (O2) was (91.33 ± 9.45) mmHg,pa (CO2) was (40.13 ± 3.72)mmHg,pH was 7.39 ± 0.03.The breathing,heart rate,oxygen saturation and arterial blood gas in both groups after treatment were improved significantly compared with those before treament,and the differences were statistically significant (all P ＜ 0.05);after treatment for 12 h,the heart rate,breathing,oxygen saturation and arterial blood gas of HHFNC group were improved more than those of the nCPAP group,and the differences had statistical significance (all P ＜ 0.05).The duration of non-invasive ventilation was (45.88-± 6.49) hours in HHFNC group,and (49.33 ± 8.99) hours in nCPAP group,so there was no difference between the 2 groups (t =1.254,P =0.219).There was 12 cases (80.0％) of mild complication in nCPAP group and 5 cases(29.4％) in HHFNC group,while the incidence rate of invasive ventilation use was 3 cases(20.0％) in nCPAP group and 1 case (5.9％) in HHFNC group.Conclusion The efficacy and security of HHFNC on moderate and severe bronchiolitis are better than those of nCPAP,and it is recommended for clinical application widely.
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Infection rate was high in children with hematologic cancers and febrile neutropenia,especially the number of drug-resistant bacteria showed an increasing trend.Because of defects of the immune function,the typical symptoms,pathogenic bacteria and infection was not clear in these children,the fever may be the only sign of serious underlying infection,mortality related with infection was high,so it was important to evalute the clinical risk accuratly and select antibiotic properly.
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ObjectiveTo explore the clinical features and factors inlfuencing the prognosis of childhood non-Hodgkin's lymphoma (NHL).MethodsPathologically diagnosed 78 pediatric patients with NHL and treated in the Afifliated Hospital of Qingdao University from January 2004 to August 2013 were collected and analyzed. Patients were grouped according to age, sex, tumor size, immunologic classiifcation, B-symptoms, LDH, hemoglobin and clinical staging. The 5-years event-free survival rate (EFS) were calculated and analyzed by Kaplan-Meier method, and the difference of the survival rate between groups were com-pared. Using Cox proportional hazards model, we analyzed the possible factors that might inlfuence 5-years event-free survival rate EFS , such as age and clinical staging. TheOR value and the 95%CI were calculated.ResultsAmong the 78 cases, median age of onset is 7 years old, male to female ratio is 2.90:1, there are 25 cases of T-cell type and 53 cases of B-cell type. According to pathological types,Burkitt lymphoma is the most common (34.6%), followed by T-lymphoblastic lymphoma (20.5%), diffuse large B-cell lymphoma (11.5%). According to the St. Jude malignant lymphoma staging system, there are 2 cases in stage I, 9 in stageⅡ, 35 in stageⅢ and 32 in stageⅣ. Swelling of periphery lymph node (80.7%) was observed as initial symptom in 26 cases of lymphoblastic lymphoma. Among 45 cases of mature B-cell tumor, the main clinical feature including abdominal cavity and gingival were observed in 27 cases of Burkitt lymphoma. Among the 73 cases received treatments, 66 cases (90.5%) attained CR (complete remission) and 4 cases (5.5%) attained PR (partial remission) by cytology and radiographic assessment after two course of combined chemotherapy, 2 cases (2.7%) rapidly relapsed after the remisson of one course treatment, 1 case (1.3%) appeared the central nervous system inifltration in the chemotherapy. With median follow-up time of 42 months, the 5-year EFS of the 73 cases was (67.0+5.5)%. Single factor analysis showed that B-symptom, LDH, and clinical staging were signiifcantly correlated with prognosis (P0.05). Multiple factor analysis showed that LDH and clinical staging inlfuenced the prognosis (OR=3.34,95%CI 2.275?10.683,P<0.01;OR=4.354,95%CI 1.519?12.475,P<0.01) .Conclusionclinical features of childhood NHL are variable. LDH and clinical staging at primary diagnosis are important factors affecting the prognosis.
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Objective To assess the effect of heat shock protein 70 of BCG (BCG HSP70) gene transfection on tumorigenicity and immunogenicity of murine lymphocytic leukemia cells (L1210). Methods BCG HSP70 gene was transfected onto the surface of murine lymphocytic leukemia cells (L1210) by lipofectamine 2000. And then the positive clone (L1210-HSP70) highly expressing HSP70 was selected as the tumor vaccine to study the tumorigenicity experiments in nude mice and syngeneic mice, the therapeutic experiments, and the immunoprotective effects. Results The expression of BCG HSP70 on the L1210 cells surface was detected, and the L1210-HSP70 cells had the same tumorigenicity as the parental L1210 cells did. Tumorigenicity experiments in syngeneic mice:In L1210-HSP70 group, tumor growth was slow or without the formation of tumor. As compared with L1210 group and L1210-neo group the mice survival time was signiifcantly prolonged, showing a marked stimulating effect on L1210 specific Th1 cells,. Tumor-bearing mice showed complete coagulation necrosis and abundant CD8+T lymphocyte inifltration (P<0.05). The tumor vaccine of L1210-HSP70 cells had the antitumor therapeutic efifcacy and immune protection effect, demonstrating that the tumor growth was signiifcantly inhibited, tumor diameter was markedly reduced and the survival time of tumor-bearing DBA/2 mice was further prolonged. Conclusions BCG HSP70 gene transfection could effectively improve the immunogenicity of tumor cells, activate speciifc T cells and enhance the anti-tumor immunity in vivo. Meanwhile, the host anti-tumor immunity could be enhanced.
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Treatment of Langerhans cell histiocytosis (LCH) needs to be tailored for each individual patient according to LCH classiifcation currently. Single-system LCH (SS-LCH) has an excellent prognosis. However, there is a poor prognosis in multisystem LCH (MS-LCH) with risk organs (RO) involvement and refractory or recurrent LCH (Re-LCH). The prognosis of MS-LCH with RO involvement and Re-LCH has been improved markedly accompanying with progress of chemotherapy in recent years. The 5-year survival rate of MS-LCH reached above 80%, and the effective rate of Re-LCH reached above 60% after chemotherapy. Re-LCH can be cured by hematopoietic stem cell transplantation.
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Objective To culture the acute leukemia cells in vitro,and to prepare cancer vaccine expressing heat shock protein 70 (HSP70) of Bacille Calemette-Geérin(BCG) onto the cell surface,so as to study its anti-tumor effect and mechanism.Methods Acute myeloid leukemia (AML) cells were cultured in a serum-free Stemspan(H) culture supplemented with cytokines [stem cell factor(SCF),flt-3 ligand (FL),interleukin (IL)-3 and IL-6] in vitro.And B-lineage acute lymphoblastic leukemia (B-ALL) cells were cultured in a Iscove modified medium(IMDM) culture supplemented with cytokines (SCF,FL,IL-3 and IL-7) in vitro.Cellular morphology was observed by the microscopy and immunophenotype determination was used to verify the biological characteristics of acute leukemia cells after culture.Lipofectamine 2000 was used to transfect the pDisplay-HSP70 plasmid into acute leukemia cells.The expression of HSP70 on the cell surface was detected by fluorescene microscope.Then the immunogenicity of the leukemia cells expressing HSP70 were detected.The experimental groups were divided into 3 subgroups:the wide-type acute leukemia cells (wt-LC group),the pDisplay-leukemia cells (pDisplay-LC group),and the pDisplay-HSP70-leukemia cells (HSP70-LC group),respectively.The leukemia cells in different groups were cultured with autologous peripheral blood T cells for 72 hours.The proliferation indices of T cells were assayed by carboxyfluorescein diacetate succinimidyl ester (CFSE)-staining method,and the contents of interferon-γ(IFN-γ) were tested by enzyme-linked immunosorbent assay (ELISA).The leukemia cells in different groups were cultured with autologous peripheral blood T cells,and after 6 days,the fresh acute leukemia cells were added [in the different ratios of cytotoxicity T lymphocyte (CTL):leukemia cells were 10 ∶ 1,20 ∶ 1,40 ∶ 1 and 80 ∶ 1] and continued to be cultured for another 12 hours.Cytotoxicity assay was measured by lactate dehydrogenase (LDH) release.Results After short term culture in vitro,the leukemia cells were in colony-like suspension and maintain the proliferation characteristics were maintained.The cell proliferation was rapidly cultured for about 10 days and then was gradually slowed down.But there was no difference between the day 10 and day 0 in the expressions of CD13 and CD33 in fifteen cases of AML cells (P ＞ 0.05).Equally,there was no difference between the day 10 and day 0 in the expressions of CD19,CD10 and CD22 in fifteen cases of B-ALL cells (P ＞ 0.05).After BCG HSP70 gene transfection,the yellow-green fluorescence on the leukemia cells surface was observed under the confocal microscope.Detection of the immunogenicity:(1) Autologous T cell proliferation:the most significant T cell proliferation was observed in the group of HSP70-transfected leukemia cells (t =17.89,19.58,all P ＜0.05).There was no difference between the wt-LC group and pDisplay-LC group (P ＞ 0.05).(2) The contents of cytokines:the IFN-γ level in the group of HSP70-transfected leukemia cells was higher than those of wide-type acute leukemia cells and the pDisplay-transfected ones (t =24.72,24.81,all P ＜ 0.05).(3) Cytotoicity of CTL:the killing rate in HSP70-transfected leukemia cells was significantly higher than those of wide-type acute leukemia cells and pDisplay——transfected ones(F =13.66,P ＜ 0.05).And with the increase of the ratio from 10 ∶ 1 to 80 ∶ 1,the inhibiting activity of CTL in the HSP70-LC group was raising(F =19.69,P ＜ 0.05).Conclusions Fresh acute leukemia cells can be successfully cultured in vitro.Short-term culture can significantly increase the number of leukemia cells,but has little effect on surface antigen expression.So,the biological characteristics of the leukemia cells can be maintained.The leukemia cells vaccine expressing BCG HSP70 onto its surface was successfully prepared,and gene transfection of BCG HSP70 can significantly enhance the immunogenicity of leukemia cells.
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The crowd were divided to 3 groups according to glucose tolerance:normal glucose tolerance,impaired glucose regulation,and new type 2 diabetes,the data was analyzed.The result showed that as the severity of abnormal glucose metabolism,the serum GGT gradually advanced,early insulin secretion index descended,The levels of resistance index,fasting and postprandial blood sugar,and glycosylated hemoglobin all raised.The serum GGT and early insulin secretion index was inversely correlated.The higher GGT level was an independent risk factors of abnormal glucose metabolism.
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Objective To explore the change of thyroid hormones and leptin at hyperuricemia (HUA)/gout.Methods A total of 96 primary gouts,65 HUAs,and 59 healthy examiners was selected.Height,weight,blood pressure,renal function,serum uric acid(SUA),glucose,lipid profiles,insulin,thyroid hormones were measured after an overnight fast.Results (1) The prevalence of subhypothyriodism at gout and HUA was 7.29％ and 15.38％,respectively.They were higher than that at healthy subjects.(2) Body mass index (BMI),systolic blood pressure (SBP),triglyceride (TG),cholesterol (CHO),thyroid stimulating hormone (TSH),fasting insulin (FINS),homeostasis model assessment of insulin resistance (HOMA-IR),and serum leptin level were increased remarkably at gout/hyperuricemia relative to control group,whereas,free thyroid hormone (FT4) was decreased.(4) In the gout and hyperuricemia groups,TSH was used as the dependent variable for the linear multivariate regression analysis,the results showed that sex,age,BMI,SUA,FT4,HOMA-IR,and Leptin were included in the regression equation of TSH (βwere-0.27,0.832,0.946,0.198,-0.942,0.895,and 0.650,respectively).Conclusions The prevalence of subhypothyroidism in primary gout/hyperuricemia was increased.Female,age,BMI,SUA,FT4,HOMA-IR,and leptin were the independent risk factors.Insulin resistant and leptin played the media roles in the gout/HUA and hypothyroidism.
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Objectives To prepare the HL-60 cell vaccine expressing heat shock protein 70 (HSP70) of Bacille calmette-Guérin (BCG), so as to study its anti-tumor effect and mechanism. Methods The whole BCG HSP70 gene was amplified from BCG genome by polymerase chain reaction (PCR) and sub-cloned into the polyclone endonuclease sites in pDisplay. The recom-binant vector of pDisplay-HSP70 was verified by sequencing. Then the HL-60 cell vaccine expressing the protein onto the cell surface was prepared by lipofectamine transfection. To detect the immunogenicity of HL-60 cells expressing HSP70, the test groups were divided into three subgroups, HL60-wt, HL60-pDisplay, and HL60-HSP70 respectively. Each group was cultured with peripheral blood T cells for 72 h, then the proliferation indices of T cells were assayed by CFSE-staining method, and IFN-γwere tested by enzyme-linked immunosorbent assay (ELISA). The HL-60 cells of different groups were cultured with peripher-al blood T cells for 6d. The wild-type HL-60 cells were added and co-cultured for another 12h. Cytotoxicity assay was measured by LDH release. Results (1) The fragment of BCG HSP70 was consistent with the theoretical value. DNA sequencing showed that the recombinant vector of pDisplay-HSP70 was correctly constructed. (2) BCG HSP70 expressed onto the HL-60 cells sur-face. (3) Detection of the immunogenicity: ①The most significant T cell proliferation was observed in the group of HSP70-transfected HL-60 cells (P0.05).②The contents of IFN-γof the HSP70-HL60 group was the highest.③The inhibiting activity of CTLs on HL-60 cells in the group of HSP70-transfected HL-60 cells was more significant than that of wide-type and pDisplay--transfected HL-60 cells. And with the increase of the E:T ratio, the inhibiting activity of CTLs in the HSP70-HL60 group was rising. Conclusions The recombinant eukaryotic expression vector (pDisplay-HSP70) of BCG HSP70 was successfully constructed. And the HL-60 cell vaccine expressing BCG HSP70 onto its surface was successfully prepared. The results showed that gene transfection of BCG HSP70 could significantly enhance the immunogenicity of HL-60 cells.
ABSTRACT
Objective To analyze the outcome of childhood B-cell acute lymphoblastic leukemia treated (ALL) with SCMC-ALL-2005 protocol. Methods Newly diagnosed B-cell ALL from May 1, 2005 to April 30, 2009 in ifve hospitals were treated and followed up according to SCMC-ALL-2005 protocol. Results A total of 601 cases with newly diagnosed B-cell ALL were enrolled. Among them, 539 cases (89.68%) were followed up until September 30, 2011. In 601 patients, there were 284 low-risk cases (LR group), 231 moderate-risk cases (MR group) and 86 high-risk cases (HR group) which were treated with SCMC-ALL-2005 protocol. The total complete remission rate during the period of induction was 98.84%and 7 cases did not achieve complete remission. The median time of the ifrst event occurring was 35 months (2.94 years). Among 539 cases completing follow-up, 403 cases (74.77%) completed treatment including 223 cases (86.43%) in LR group, 150 cases (73.17%) in MR group and 30 cases (39.47%) in HR group. The rate of cases completing treatment was signiifcantly different among three groups (P=0.001). The completion rate was highest in LR group and lowest in HR group. The 3-year overall survival (OS) rate was (83.3±1.8)%, and the 3-year EFS (event-free survival) rate was (79.2±1.9)%using a Kaplan-Meier method. The 5-year OS rate was (79.5±3.3)%, and the 5-year EFS rate was (70.9±3.7)%. There were signiifcant differences in 3-year EFS rate and 5-year EFS rate among three groups (P<0.05). Conclusions Childhood B-ALL treated with SCMC-ALL-2005 protocol achieved a better therapeutic effect and prognosis. The multi-center collaborative research is useful for the standard treatment of ALL.