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Objective:To select human chronic myeloid leukemia (CML) cell line K562 as the experimental object, and use lentivirus mediated CRISPR/Cas9 gene editing technology to construct a stable CML cell line K562/TCRP1-KO that knocks out the tongue cancer resistance related protein 1 (TCRP1) gene; and through functional tests such as cell proliferation, apoptosis, and drug sensitivity, compare the phenotypic differences between K562/TCRP1-KO and control cells (K562/cas9-CTL), and preliminarily explore the possible mechanism of TCRP1 gene involvement in the pathogenesis of CML.Methods:The small guide RNA (sgRNA) targeting TCRP1 was designed at a specific location. After annealing, the oligonucleotide fragments were recombined with the linearized Cas9 expression vector, and the lentivirus packaging system was transfected into 293T cells. The purified virus was collected and infected with K562 cells. Positive polyclons were screened for puromycin pressure, and monoclonal K562/TCRP1-KO was further screened by limited dilution method. Stable cell lines were successfully knocked out by sanger sequencing and Western blot detection; Simultaneously, K562 cells transfected with lentiCRISPR vector were constructed as control cell lines (K562/cas9-CTL); Using cell counting method, cell counting kit 8 (CCK8) method, imatinib (IM) gradient dilution method, and flow cytometry cell proliferation, drug sensitivity, and apoptosis analysis were performed on K562/TCRP1-KO and K562/cas9-CTL, respectively.Results:The sgRNA-Cas9 recombinant plasmid vector for TCRP1 knockout was successfully constructed, and after transfection into 293T cells, TCRP1 knockout monoclonal cell lines were successfully screened using limited dilution method. Compared with K562/cas9-CTL cells, the proliferation ability of K562/TCRP1-KO cells was significantly reduced, IM drug sensitivity was significantly enhanced, and the process of cell apoptosis was significantly accelerated (all P<0.05). Conclusions:A CML cell line with TCRP1 knockout was successfully constructed using CRISPR/Cas9. TCRP1 may act as a cancer related gene to affect the proliferation, IM resistance, and apoptosis process of CML cells.
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Objective:To analyze the public demands for information about congenital birth defects in “Baidu zhidao” based on word frequency retrieval.Methods:Based on discussion between obstetrics and gynecology experts and epidemiological experts, the key words related to congenital birth defects were determined and the search strategy was formulated. Python 2.7 was used for web crawler search. Questions related to congenital birth defects were obtained on the “Baidu zhidao” platform, and then the R 4.0.2 software was used to process the data, complete the semantic analysis of keywords and statistical analysis of word frequency, and draw word cloud graph and polar chart to describe the key results.Results:A total of 16668 non-repetitive questions were retrieved from “Baidu zhidao” platform, and the frequency of semantic words was 15 371. Among them, 35.02% were the names and symptoms of congenital birth defects. In addition, the frequency of congenital heart disease was the highest (26.09%). The results of subject analysis of key words of birth defects showed that the average word frequency of diagnosis and treatment semantic words (49.55) was significantly higher than that of etiology and prevention semantic words (12.47). In addition, the key words of examination, cause, treatment, development and heredity were more frequently used in the semantic words related to the seven types of systemic malformations.Conclusion:The public in China has a high demand for information on congenital birth defect related diseases, and their causes, prevention and treatment, especially congenital heart disease.
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Objective:To evaluate the long-term health-related quality of life (QOL) in pediatric patients with acute leukemia after hematopoietic stem cell transplantation (HSCT) and to analyze potential influence factors.Methods:Patients with acute leukemia aging 8-18 years who received HSCT in the Hematology Oncology Center of Beijing Children′s Hospital from June 2009 to June 2012 with more than 80 months survival postoperatively were recruited.All of them were subjected to a short-term QOL survey in 2013.PedsQL? Transplantation Module 3.0 in Chinese mandarin version was completed.QOL data and influence factors were analyzed.Results:Forty-one patients completed the questionnaires, involving 32 males and 9 females with the mean age of(14.29±2.72) years.The mean scores of overall long-term QOL after HSCT were above 75 (total scores: 100), which was above the average.The age, disease status before transplantation, donor sources, post-transplant complications and the parental education level were the influential factors for the long-term QOL in pediatric patients with acute leukemia at post-HSCT, which could affect a certain dimension in QOL.Conclusions:The overall long-term QOL of pediatric patients with acute leukemia who survived for more than 80 months at post-HSCT is acceptable, which is significantly better than the short-term QOL after 4 months of HSCT.The age, disease status before transplantation, donor sources, post-transplant complications and the education level of parents could affect a certain dimension of QOL.
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Objective:To understand the relationship between joint bleeding and joint disease in hemophilia children, and to provide a theoretical basis for clinical treatment and prognosis.Methods:The patients with severe hemophilia A between 1 and 7 years old and with relevant nodal bleeding records were selected.All the patients admitted in Beijing Children′s Hospital, Capital Medical University, and Chengdu New Century Women′s and Children′s Hospital since June 2016 to January 2017.All the joint bleeding of each child was taken as the study joint, and the joint bleeding was collected during the last 3 months.The joints were assessed by using ultrasound, X-ray, magnetic resonance imaging(MRI) and Hemophilia Joint Health Score (HJHS) scoring systems.The correlation analysis was conducted between the joint bleeding, ultrasound, X-ray, MRI and HJHS scores.The correlation analysis was conducted for baseline ultrasound, X-ray, MRI and HJHS scores.Results:(1) There were 18 patients enrolled.The mean age was (5.6±1.8) years old.There were 30 joints bleeding in the observation period in total, with the annul median joint bleeding times of 4 (4-16 times), and the annul median target joint bleeding times of 8 (4-16 times). (2) Joint bleeding times of was correlated with ultrasound and X-ray evaluation ( r=0.390, P=0.033; r=0.517, P=0.008), and not correlated with HJHS or MRI(all P>0.05). (3) There was significantly positive correlation among ultrasound, X-ray, HJHS and MRI [ r=0.815(ultrasound vs.X-ray), r=0.510(ultrasound vs.HJHS), r=0.812(ultrasound vs.MRI), r=0.666(X-ray vs.HJHS), r=0.911(X-ray vs.MRI), r=0.781(HJHS vs.MRI), all P<0.01]. (4) There was no correlation between times and assessment for joints whose ultrasound and /or MRI in joints with abnormal ultrasound and /or MRI evaluation( P>0.05). Conclusions:The results of joint bleeding and joint evaluation are inconsistent.Joint bleeding can not truly reflect the situation of joint diseases.The assessment of hemophilia should include comprehensive evaluation of joint structure, function, activity ability and other aspects to guide the treatment of haemophi-lia children.
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Objective To retrospectively analyze the clinical efficacy,safety and influencing factors of radiotherapy in children with stage Ⅳ high-risk neuroblastoma (HR-NB).Methods A total of 120 children with HR-NB who were diagnosed and treated with local radiotherapy according to the BCH-HR-NB-2007 protocol in the Oncology Department of Beijing Children's Hospital from January 2014 to December 2017 were enrolled.Among them,56 children were male and 64 female with a median age of 43 months (9 -148 months).The treatment protocol consisted of 4 cycles of CAV chemotherapy,3 cycles of CVP chemotherapy,surgical resection after 4 cycles,autologous hematopoietic stem cell transplantation after 7 cycles,local radiotherapy at a dose of 15.0-30.6 Gy for 82 cases of primary tumors and 38 cases of primary and metastatic tumors,followed by 13 cis-retinoic acid as maintenance therapy.The entire treatment protocol endured for approximately 18 months.Results The median follow-up time was 21 months.The 3-year local control rate was 84.4％.Before radiotherapy,the 3-year event-free survival rate was 78.4％ in children without metastases,significantly higher compared with 30.4％ in the residual group (P=0.003).The 3-year event-free survival rate was 66.1％ in patients who underwent radiotherapy within 6 months after surgery,significantly higher than 50.6％ in their counterparts receiving radiotherapy at 6 months or more after surgery (P=0.018).Among the children with residual metastases before radiotherapy,the progression rate in children who did not receive radiotherapy was 66.6％,significantly higher compared with 20.0％ in those receiving radiotherapy (P=0.001).All patients had no radiation-related adverse reactions in the liver,kidney and heart,etc.The incidence rate of grade Ⅲ-Ⅳ myelosupression was 24.5％ at 1 week post-radiotherapy,and 8％ at 2 weeks after radiotherapy.Conclusions Radiotherapy yields definite clinical efficacy in the local control of children with stage Ⅳ HR-NB.Early radiotherapy after surgery and radiotherapy for the metastatic lesions can improve the clinical prognosis.No vital organ injuries are observed during the short-term follow-up.At 2 weeks after radiotherapy,the myelosupression is gradually restored.
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Objective@#To detect the arthropathies on no bleeding history joints in pre-school hemophilia A children in order to provide evidence for further prevention and control of joint disease in children with hemophilia A.@*Methods@#This study was a cross-sectional study based on China Hemophilia Individualized prophylaxis study (CHIPS). The basic data of outpatients with hemophilia in Beijing Children's Hospital and Chengdu Women's and Children's Central Hospital between August 2016 and June 2017 were collected and a three-month follow-up was conducted. The target joints (six joints of bilateral elbows, knees and ankles) of thirty-four children aged 1-7 years old with severe hemophilia A were examined by ultrasound, X-ray and joint function examination (4-7 years old, hemophilia joint health score (HJHS)). To find out whether there are arthropathies in patient's joints with no bleeding history and analyze the relevant factors by chi-square test, rank sum test and other statistical methods.@*Results@#There were 32 analyzable cases with 112 no-bleeding history target joints, 42.9% (48/112) were elbow joints. Arthropathies were revealed in 34.8% (39/112) of them by joint structural and functional examination and 46.2% (18/39) were ankles (χ2=8.379, P=0.015) . Ultrasound showed abnormalities in 18.3% (20/109) joints, X-ray showed abnormalities in 3.8% (3/79) joints and HJHS showed abnormalities in 25.3% (20/79) joints. There was no correlation between ultrasound and HJHS (r=0.015, P=0.895), no correlation was found between X-ray and HJHS (r=-0.101, P=0.390) either, which suggested that joint structural and functional examination could not replace each other. The related risk factors of arthropathies in this group were >4.91 years old (OR=3.917, 95%CI:1.610-9.528) and combining with target joint (OR=3.530, 95%CI:1.316-9.465).@*Conclusions@#Detecting the joint structure and function on no bleeding history joints in pre-school hemophilia A children could reveal the arthropathies and majority of them were ankle arthropathies. Joint structural and functional examinations could not replace each other. For patients more than 5 years old and those with target joints, the joints with no complaint of bleeding should be examined regularly to reveal the arthropathies in time.
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Objective To improve substantially the prediction accuracy of histopathology subgroup through combined analysis of clinical and biological features,and accordingly build a decision tree to predict the histopathology subgroup in intermediate and high - risk neuroblastoma. Methods A total of 62 intermediate and high - risk neuro-blastoma patients were included retrospectively in this study,who received chemotherapy and surgery at Beijing Chil-dren's Hospital (BCH),Capital Medical University between January 2015 and January 2017. The fin-dings of urinary vanillylmandelic acid (VMA),and homovanillic acid (HVA),serum neuron - specific enolase (NSE),lactate dehy-drogenase (LDH)and ferritin,ultrasound,CT,MRI,positron emission tomography - computed tomography (PET -CT),bone marrow aspiration and biopsy,MYCN gene,and histopathology were collected and analyzed retrospectively. Statistical analysis was performed by using SAS 9. 4. Univariate and multivariate Logistic regression analysis were con-ducted to select potentially useful characteristics for prediction. Based on the results of Logistic regression analysis,a classification tree was developed to predict histopathology subgroup. Results To identify the characteristics related to histopathology,tumor markers and six clinico - pathologic factors were evaluated by univariate analysis. The results showed that unfavorable histopathology(UH)was more frequently associated with bone marrow metastasis,older age,as well as higher serum NSE,ferritin and LDH levels. The result of multivariate analysis showed that age and NSE were significant independent predictors of histopathology. The adjusted odds ratio(OR)of NSE and age was 33. 2 and 13. 0, respectively. The area under the receiver - operating - characteristic (AUC)of the prediction mo-del was 0. 889. The sensitivity and specificity were 91. 90％ and 76. 00％,respectively. Furthermore,to provide a visua-lization of the sig-nificant predictors found by Logistic regression analysis,a decision tree was developed for predicating of histopathology. Conclusions Age and NSE are significant independent predictors of histopathology subgroup. The decision tree based on age and NSE can help to predict the histopathology subgroup in intermediate and high - risk neuroblastoma effective-ly.
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<p><b>OBJECTIVE</b>To determine the percentage of hypercholesterolemic patients who had met the criteria as total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C), defined by the Chinese National Recommendations for Prevention and Treatment of Dyslipidemia.</p><p><b>METHODS</b>Adult patients with hypercholesterolemia, who had been receiving the same lipid-lowering therapy for at least 2 months, were enrolled. Lipid levels were determined at the time of enrollment, to assess whether the patients' lipid levels had reached the criteria for treatment. Patients' cardiovascular risk factors and lipid-lowering treatments were also collected.</p><p><b>RESULTS</b>One hundred and eighty patients with mean age of 65.8 were studied. Of these, 6.7% had no risk factors and no definite disease of atherosclerosis (low-risk group), 65.5% had risk factors but no documented atherosclerosis (high-risk group), and 27.8% had established atherosclerosis diseases or diabetes mellitus. Overall, only 44% of patients achieved both TC and LDL-C target levels. The success rates were higher among low and high-risk groups than that among patients with atherosclerosis or diabetes mellitus. The relationship between four different lipid-lowering drug therapies and successful patient outcome was also investigated. The success rates were 51.8% for simvastatin, 42.9% for pravastatin, 31.6% for fluvastatin, 12.5% for other drugs respectively.</p><p><b>CONCLUSION</b>More than half of the hypercholesterolemic patients receiving lipid-lowering therapy had not achieved TC and LDL-C target levels. Data from this study indicated that a significant gap still existed between dyslipidemia prevention principles and clinical practices, suggesting that more aggressive treatment of dyslipidemia is needed.</p>