ObjectiveTo investigate the clinical phenotypes of familial hypercholesterolemia (FH) caused by exon 13 A606T mutation in low deusity lipoprotein receptor.MethodsClinical data of the suffered family were collected and analyzed,as well as measurement of perivascular intima-medial thickness and follow-mediated-dilation function by ultrasonography.ResultsThere were totally 11 sufferers including 4 males and 9 females,aged 8-90 years,with 2 homozygotes and 9 heterozygotes.Among them, one homozygote showed angina pectoris and hematuria,both homozygotes had skin xanthomata.TC,TG,LDL-C and HDL-C were(7.39 ± 1.30) mmol/L,(0.93 ± 0.36) mmol/L,( 11.76 ± 1.10) mmol/L and ( 1.22 ±0.17) mmol/L,respectively.The left/right sided intima-medial thickness of the common,internal,external and bulb carotid artery were ( 1.15 ±0.45) mm/( 1.30 ±0.60) mm,(0.82 ±0.30) mm/( 1.00 -0.66)mm,(0.77 ±0.28) mm/(0.78 ±0.30) mm and ( 1.40 ±0.59) mm/( 1.46 ±0.71 ) mm respectively.The brachial artery flow mediated dilation rate was (4.85 ±4.80)％.Echocardiography revealed 2 patients with cardiac valvular disease and 3 with atrium septum aneurysm. ConclusionFH patients show a variety of phenotypes incuding extraordinary hypercholesterolemia,subcutaneous xanthomata and premature coronary heart disease.

Objective To investigate low density lipoprotein receptor (LDLR)gene mutation in familial hypercholesterolemia (FH) patients. Methods The proband was given clinical diagnosis of homozygous FH based on marked features and blood lipid tests results. After apoB100R3500Q mutation was excluded, the promoter region and all of the 18 exons of LDLR gene were amplified by touch-downpolymerase chain reaction (PCR). The PCR products were analyzed by single-strand conformationalpolymorphism (SSCP). The PCR products with abnormal single strands were sequenced directly. Thesecondary structures of the mutational and wild type proteins were analyzed and compared byANTHEPROT5.0, and then the tertiary structures of the mutant and wild type LDLR were predicted atSWISS MODEL homepage online. Results A homozygous mutation A606T at exon 13 of the patients wasfound by SSCP and confirmed by DNA sequencing. GOR Ⅰ method in ANTHEPROT5.0 indicates that therandom coils and turns would replace some helixes at the mutation site. The online prediction from theSWISS MODEL homepage indicates the backbone structure of the mutant LDLR has no difference from thewild type one. Conclusion The results suggest the A606T mutation of LDLR gene is the cause of the FH inthis pedigree.

Objective To evaluate the clinical significance of minimal residual disease (MRD)detecion in ALL-B of children by flow cytometric (FCM).Methods 52 cases of children with ALL-B were performed bone marrow MRD by FCM analisis after induction therapy,3 moths therapy,and 6 moths therapy.After that,MRD detection was performed every 6 months. According to disease risks, three group were categorized,standard risk (SR),imidiete risk (IR) and high risk(HR).Results After 6 months,SR groups MRD positive cases were 4/21(19 ％),IR groups MRD position cases were 8/23 (35 ％),HR groups MRD position cases were 5/8 (63 ％).9 cases relapsed in all 52 patients.There were significant differrence in replased rate between the positive and negtive MRD (P＜0.001). Conclution The dynamic detection of MRD by FCM can be used to evaluate the therapeutic effect and prognosis of children with ALL-B. It is also useful in adjusting treatment strategy and for following up in children with ALL.

Objective:To explore the clinical characteristics and prognostic factors of children with relapsed acute lymphoblastic leukemia (ALL).Methods:The clinical data of 52 children with relapsed ALL in Children's Hospital of Shanxi Province from January 2010 to April 2019 were retrospectively analyzed. The clinical characteristics of the children were summarized and the prognostic factors after recurrence were analyzed.Results:Till May 1, 2019, 5 out of 52 children gave up treatment after diagnosis and were lost to follow-up. For the remaining 47 children with successful follow-up, the median age at initial diagnosis was 60 months (11-168 months), the median time from initial diagnosis to relapse was 21 months (2-112 months), the median follow-up time was 5.5 months (1.0-69.0 months), and the 2-year overall survival (OS) rate after relapse was 31%. Nine patients accepted allogeneic hematopoietic stem cell transplantation after the second time complete remission, the median time from diagnosis to transplantation was 4.5 months (3.0-7.0 months), and the median follow-up time was 22 months (4-69 months). The 2-year OS rates in relapsed children with white blood cell count < 50×10 9/L and ≥ 50×10 9/L at initial diagnosis were 39% and 13%, respectively (χ 2=5.623, P=0.018). The 2-year OS rate after relapse in standard-risk, intermediate-risk and high-risk groups were 72%, 31% and 8%, respectively (χ 2=10.068, P=0.007). The 2-year OS rate after relapse in very early relapse, early relapse and late relapse groups were 0, 33% and 79%, respectively (χ 2=30.066, P < 0.01). The 2-year OS rate after relapse in chemotherapy with or without radiotherapy group, transplantation group and irregular treatment group were 57%, 89% and 0, respectively (χ 2=26.885, P < 0.01). Cox multivariate analysis showed that relapse time was the independent risk factor affecting the prognosis of children with relapsed ALL ( HR=0.340, 95% CI 0.146-0.789, P=0.012). Compared with the transplantation group, the risk of death in the chemotherapy with or without radiotherapy group and the irregular treatment group was significantly higher ( HR=12.313, 95% CI 1.266-119.758, P=0.031; HR=20.699, 95% CI 2.230-192.129, P=0.008), suggesting that hematopoietic stem cell transplantation is a protective factor for the prognosis of children with relapsed ALL. Conclusions:The relapse of ALL in children mainly happens in very early and early time. The main part of relapse is bone marrow, and there are many high-risk patients at initial diagnosis. The risk group at initial diagnosis, white blood cell count at initial diagnosis, relapse time, and treatment after relapse are the risk factors affecting the prognosis, and the relapse time and hematopoietic stem cell transplantation are the independent prognostic factors.

Objective:To evaluate the short-term efficacy and the improvement of quality of life of enzyme replacement therapy (ERT) with Imiglucerase on children with Gaucher disease(GD) through the same time monitoring.Methods:Six children diagnosed as GD who were treated by ERT with Imiglucerase in the Department of Hematology of the Children′s Hospital of Shanxi Province from May 2019 to May 2020 were recruited.Every 3 months, the sizes of the liver and spleen was palpated, the change of bone pain was recorded, and the haematological index was examed.The volumes of the liver and spleen at 1-year treatment were measured by CT.Bone involvement was examined by magnetic resonance imaging (MRI). In addition, the body weight, height, and the 36-Item Short Form Survey (SF-36) were measured and compared with pre-treatment levels.These data were analyzed statistically by SPSS 25.0 and the difference between pretherapy and post-treatment was compared by paired t test. Results:Six children diagnosed as GD received ERT with Imiglucerase.No adverse events were reported.Decreased volumes of the liver and spleen, and increased hemoglobin level and platelet count were detected after 3-6 months of ERT.After 1 year of ERT, hemoglobin level significantly increased compared with pre-treatment level ( t=4.200, P=0.008). Although the platelet count increased at 1-year ERT, it was comparable with pre-treatment level ( t=2.260, P=0.073). The volumes of liver and spleen decreased by (22.10±15.28)% ( t=2.725, P=0.042) and (47.10±18.42)% ( t=3.162, P=0.034) after 1 year of ERT, respectively.During the first year of ERT, the height and weight increased (6.17±2.86) cm ( t=5.286, P=0.003) and (4.08±2.01) kg ( t=4.975, P=0.004), respectively.SF-36 score increased significantly from (489.35±103.99) points to (632.75±73.34) points ( t=5.740, P=0.002). After 1 year of ERT, 1 patient still had bone pain, and 2 cases were worse in bone MRI, which may be attributed to the short period of follow-up and insufficient dose, and another 3 had no change in bone MRI. Conclusions:ERT ameliorates GD-associated anemia, organomegaly and growth retardation, and improves the growth rate of body mass and height and the quality of life in the short period.However, short-term ERT does not improve the bone disease.

OBJECTIVETo explore the effect of liraglutide on myocardial ischemia/reperfusion (I/R) injury in rats and related mechanisms.

METHODSTwenty-four male SD rats were divided into sham group, I/R injury group, and liraglutide group by table of random number (n = 8 each). Myocardial I/R injury model was established by occlusion of the left anterior descending artery for 30 min followed by 2 h of reperfusion. HE stain method was used to observe cardiomyocyte status under light microscope, myocardial tissue samples were stained with TTC to measure the myocardial infarction size, protein expression of p53 and caspase-3 was analyzed by immunohistochemical technique and Western blot respectively, xanthine oxidase method was used to detect SOD activity, Thiobarbituric acid method was used to measure the concentration of MDA.

RESULTSCompared with the I/R group, the degree of myocardial damage of liraglutide group was significantly reduced and the myocardial infarct area was significantly lower ((44 ± 8) % vs. (62 ± 8) %, P<0.05) while protein expression of caspase-3 and p53 in liraglutide group was significantly downregulated (0.19 ± 0.03 vs. 0.24 ± 0.02 and 0.27 ± 0.03 vs. 0.39 ± 0.04, P<0.05). SOD activity was significantly increased and MDA concentration was significantly decreased (74.20 ± 11.10 vs. 44.04 ± 14.30 and 4.41 ± 1.07 vs. 8.72 ± 2.20, P<0.05) in liraglutide group compared to I/R group.

CONCLUSIONLiraglutide protects myocardium against I/R injury possibly through reducing cardiomyocytes apoptosis and oxidation.

Animals ; Apoptosis ; Caspase 3 ; Coronary Vessels ; Liraglutide ; Male ; Myocardial Infarction ; Myocardial Reperfusion Injury ; Myocardium ; Myocytes, Cardiac ; Oxidation-Reduction ; Rats ; Rats, Sprague-Dawley

Objective: To investigate the clinical characteristics and prognosis of children B-cell acute lymphoblastic leukemia (B-ALL) with TEL-AML1 fusion gene positive. Methods: Clinical characteristics, therapeutic effects and prognostic factors of 55 children B-ALL patients with TEL-AML1 fusion gene positive in Children's Hospital of Shanxi from January 2013 to June 2018 were retrospectively analyzed. Kaplan-Meier method was used to evaluate 3-year event-free survival (EFS) rate and overall survival (OS) rate. Influencing factors of EFS and OS were evaluated by using Cox regression analysis. Results: TEL-AML1 fusion gene was positive in all 55 children, and no other fusion gene positive was merged. There were 4 patients (7.3%) ≥10 years old. At initial diagnosis, 33 patients (60.0%) had hepatomegaly, 28 patients (50.9%) had splenomegaly, and 27 patients (49.1%) had superficial lymphadenectasis. There were 5 patients (9.1%) with white blood cell count ≥50×10⁹/L, and 19 patients (34.6%) had abnormalities of chromosome. All the 55 children were divided into the low risk group [36 cases (65.5%)], the intermediate risk group [18 cases(32.7%)], high risk group [1 case (1.8%)] according to Morphology, Immunology, Cytogenetics and Molecular Biology (MICM) and adjusted risk. After regular treatments, 50 patients achieved complete remission (CR) on the 15th day. The CR rate after one-course of induction therapy was 100.0%. On the 33rd day, 43 patients (78.2%) had minimal residual disease (MRD) <10^-4, 12 patients (21.8%) had MRD≥10^-4 and MRD<10^-2, 1 patient (1.8%) had MRD≥10^-3 at the 12th week. During three to six months, the negative rate of fusion gene was 61.8% (34/55). There were 3 deaths (5.5%), and one (1.8%) of them died of recurrence, and the recurrence time was 27 months from the initial diagnosis; the other 2 cases (3.6%) died of infection during chemotherapy. In 55 patients, the 3-year EFS rate and OS rate was 90.3% and 93.2%, respectively. The 3-year EFS rate and OS rate in the low risk group was 100.0% both; the 3-year EFS rate and OS rate in the intermediate risk group was 78.7% and 86.6%, respectively; the 3-year EFS rate and OS rate in the high risk group was 0 both and one died. EFS rate and OS rate in low risk group were higher than those in the intermediate risk group, and the differences were statistically significant (P < 0.05). The EFS rate was 92.0% and 0 at the 12th week MRD<10^-3 group and MRD≥10^-3 group, and OS rate was 95.0% and 0 at the 12th week MRD<10^-3 group and MRD≥10^-3 group (P < 0.05). Cox multivariate analysis showed that MRD at the 12th week was an independent risk factor influencing EFS and OS (OR= 2.971, 95% CI 1.330-6.633, P= 0.008; OR= 2.884, 95% CI 1.295-6.419, P= 0.009). Conclusions Children B-ALL patients with TEL-AML1 fusion gene positive have a low recurrence rate, high survival rate and good prognosis. Risk stratification and the 12th week MRD are the influencing factors of the prognosis.

Objective:The matria-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS) and sequencing methods were performed to assess the methodology (biochemistry methods) for identifying the Pandoraea sputorum and provide the more preferred approach to identify Pandoraea species. Methods:This paper is a study on performance evaluation of identification method. Ten lines of Pandoraea sputorum were isolated from blood cultures of inpatients were collected at Zhongnan Hospital of Wuhan University from July to October 2018 and were confirmed by the cultural characteristics, colonial morphology and Gram′s stain. Further identification was carried out by using the manual biochemical method (API 20NE), automatic biochemistry systems(BioMerieuxVITEK 2 Compact, BD Phoenix-100andThemo ARIS 2X), MALDI-TOF MS (BioMerieuxVITEK MS and Bruker MALDI Biotyper) and the sequencing methods of the 16S rRNA to identify the Pandoraea sputorum. Results:Pandoraea sputorum was non-fermented gram-negative bacteria that are non-motile, oxidase positive, and catalase positive. Ten lines of Pandoraea sputorum were identified as Achromobacter denitrificans, Alcaligenes faecalis or Cupriavidus pauculus and the accuracy rate of genus and species identification was 0 by API 20NE. Among all the samples, six lines were identified as the Pandoraea spp. with the accuracy rate of genus identification was 6/10 by VITEK 2 Compact; whereas the other four lines were identified as the Burkholderia cepacia, Sphingomonas paucimobilis, Ralstonia pickettii, or "Low Discrim" . All of these were identified as "No Identification" by Phoenix-100, which the accuracy rate of genus and species identification was 0. Seven isolates were identified by ARIS 2X as Stenotrophomonas maltophilia, Acinetobacter lwoffii, Sphingomonas paucimobilis, Pseudomonas luteola, Acinetobacter baumannii/Acinetobacter haemolyticus; whereas the other three lines were identified as rare species, thus, the accuracy rate of genus and species identification was 0. Both VITEK MS and MALDI Biotyper indicated all the isolates were Pandoraea sputorum with the accuracy rate of genus and species identification was 10/10. 16S rRNA sequencing for the 10 isolates showed they have 100% of similarity to Pandoraea sputorum by blasting with Genebank. Conclusions:Methods based on biochemical reactions often failed to accurately identify the Pandoraea sputorum to species level. MALDL-TOF MS and 16S rRNA sequencing technology identify Pandoraea sputorum efficiently and precisely enough.

Objective:To investigate the relationship between NUDT15 gene polymorphism and tolerance to treatment with 6-mercaptopurine (6-MP) in children with acute lymphoblastic leukemia (ALL).Methods:Fifty-eight children diagnosed with ALL in Shanxi Children's Hospital from January 2019 to December 2020 were recruited. All of them were treated with CCLG-ALL2018 chemotherapy regimen and the bone marrow showed complete remission. They received 6-MP oral treatment during maintenance treatment. Single nucleotide polymorphism of NUDT15 gene was detected by real-time fluorescence quantitative polymerase chain reaction. The bone marrow suppression after 6-MP treatment and 6-MP tolerance dose in patients with different NUDT15 genotypes were analyzed.Results:Among 58 patients, 3 patients had NUDT15 TT genotype, 46 patients had CC genotype and 9 patients had TC genotype. During maintenance treatment with 6-MP, the differences in leukocyte count, hemoglobin and platelet count among the three groups of patients with different NUDT15 genotypes were statistically significant (all P < 0.05). Among 58 patients, 23 (39.66%) patients had varying degrees of neutropenia after medication, including 16 cases of NUDT15 CC genotype, 5 cases of TC genotype and 2 cases of TT genotype. There was a statistically significant difference in bone marrow suppression among the three groups ( H = 29.10, P < 0.05). The dosages of 6-MP used in patients with TT, CC and TC genotypes were (10.4±8.8) mg·m -2·d -1, (41.5±1.3) mg·m -2·d -1 and (36.7±2.4) mg·m -2·d -1, respectively, and the difference was statistically significant ( F = 16.95, P < 0.05). Conclusions:Children with different NUDT15 genotypes have different tolerance to 6-MP, and NUDT15 gene polymorphism is associated with 6-MP intolerance during maintenance treatment in children with ALL, which may affect the treatment of the disease.

Objective To explore the change in 25 hydroxyvitamin D[25 -(OH)D]level in school - aged children with orthostatic hypertension (OHT). Methods Nineteen cases of school - aged children with OHT confirmed diagnosis by head - up tilt table test at the Department of Pediatric Cardiovasology,Children′s Medical Center,the Second Xiangya Hospital,Central South University,from October 2014 to February 2017,were selected as OHT group, including 17 males and 2 females,and their ages were from 7 to 14(11. 21 ± 2. 70)years old. Nineteen healthy children including 17 males and 2 females and aged 8 to 14(11. 05 ± 2. 35)years old who had a healthy examination of child care at the hospital in the same period were selected as healthy control group. In two groups of children all possible basic diseases were eliminated,such as severe liver and kidney disease,abnormal thyroid function and metabolic bone disease and/ or the long - term use of 25 -(OH)D metabolism drugs,accepted the serum 25 -(OH)D detection. Results (1)There was no significant difference in age and gender between the OHT group and the healthy control group(t = 0. 559,P > 0. 05;χ2 = 0. 000,P > 0. 05). The 25 -(OH)D levels were significantly lower in the OHT group than those in the healthy control group [(39. 62 ± 10. 65)nmol/ L vs. (64. 83 ± 10. 28)nmol/ L,t = - 7. 422,P <0. 01]. (2)25 -(OH)D levels had no correlation with age,gender,height,body mass,systolic pressure,or diastolic blood pressure (r = 0. 254,0. 047,0. 195,0. 019,- 0. 191,- 0. 184,all P > 0. 05). Taking 25 -(OH)D level as dependent variable,age,gender,height,body mass,systolic pressure,diastolic blood pressure as independent variables, multiple stepwise regression equation to predict 25 -(OH)D level was not fit. Conclusion Lower level of 25 -(OH)D may be one of the mechanisms for the onset of the school - aged children with OHT.