Expression and Clinical Significance of Exosome Derived MiR-181b-5p in Children with Acute Lymphoblastic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To explore the expression level of exosome derived miR-181b-5p in different disease stages of children with acute lymphoblastic leukemia and its relationship with clinical characteristics.@*METHODS@#Bone marrow plasma samples of 86 children with ALL were collected. Exosomes were extracted by exosome extraction kit, and RNA in exosomes was extracted by TRIzol method. The levels of miR-181b-5p in the blood plasma exosomes of the patients in the newly diagnosed group, relapse group, remission group and control group were detected by qRT- PCR. The difference of miR-181b-5p expression level in each group was compared and analyzed, and the relationship between miR-181b-5p expression level and clinical characteristics was analyzed.@*RESULTS@#The expression level of exosomal miR-181b-5p in the newly diagnosed group and the relapsed group was significantly lower than that in the remission group and the control group (P< 0.05). The expression level of exosomal miR-181b-5p in T-ALL children was higher than that in B-ALL children (P<0.05). The expression level of plasma exosomal miR-181b-5p in male children was higher than that in female children (P<0.01).@*CONCLUSION@#Exosome derived miR-181b-5p changes dynamically in the course of ALL children, and can be used as a marker miRNA to monitor disease status. Exosomes can transmit information in the tumor microenvironment and serve as a potential carrier for biomolecular targeted therapy.

Single Center Clinical Analysis of Bloodstream Infection Pathogens in Children with Acute Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the clinical features, distribution of pathogenic bacteria, and drug resistance of bloodstream infection in children with acute leukemia.@*METHODS@#Clinical data of 93 blood culture-positive children with acute leukemia from January 2015 to December 2019 in Department of Pediatrics, The Second Hospital of Anhui Medical University were analyzed retrospectively.@*RESULTS@#In these 93 cases, 78 cases were in the period of neutrophil deficiency. There were 54 Gram-negative bacteria (G-) (58.1%) found through blood culture, and the top 4 strains were Escherichia coli (15.1%), Klebsiella pneumoniae (13.9%), Pseudomonas aeruginosa (6.5%), and Enterobacter cloacae (6.5%). There were 39 Gram-positive bacteria (G+) (41.9%) detected, and the top 4 strains were Staphylococcus epidermidis (10.8%), Streptococcus pneumoniae (6.5%), Staphylococcus hemolyticus (5.4%), and Staphylococcus human (5.4%). Among 74 strains of pathogenic bacteria from acute lymphoblastic leukemia (ALL) children, there were 29 strains of G+ bacteria (39.2%) and 45 strains of G- bacteria (60.8%). While in 19 strains from acute myeloblastic leukemia (AML) patients, G- bacteria accounted for 47.4% and G+ bacteria accounted for 52.6%. In 15 ALL children without neutropenia, G+ bacteria made up the majority of the strains (66.7%). In the 93 strains of pathogenic bacteria, 13 (13.9%) strains were multidrug-resistant. Among them, extended-spectrum β-lactamases accounted for 42.9%, carbapenemase-resistant enzyme Klebsiella pneumoniae 15.4%, and carbapenemase-resistant enzyme Enterobacter cloacae strains 33.3%, which were detected from G- bacteria. While, 13.3% of methicillin-resistant coagulase-negative Staphylococci accounted for 13.3% detected from G+ bacteria, but linezolid, vancomycin, teicoplanin Staphylococcus and Enterococcus resistant were not found. The average procalcitonin (PCT) value of G- bacteria infection was (11.02±20.282) ng/ml, while in G+ infection it was (1.81±4.911) ng/ml, the difference was statistically significant (P<0.05). The mean value of C-reactive protein (CRP) in G- infection was (76.33±69.946) mg/L, and that in G+ infection was (38.34±57.951) mg/L. The prognosis of active treatment was good, and only one case died of septic shock complicated with disseminated intravascular coagulation (DIC) and gastrointestinal bleeding caused by carbapenemase-resistant enzyme enterobacteriaceae.@*CONCLUSION@#G- is the major bacteria in acute leukemia children with bloodstream infection, but the distribution of ALL and AML strains is different. G- bacteria dominates in ALL, while G+ bacteria and G- bacteria are equally distributed in AML. Non-agranulocytosis accompanied by bloodstream infections is dominant by G+ bacteria. The mean value of PCT and CRP are significantly higher in G- bacteria infection than in G+ bacteria.

Influencing Factors in the Chronicity of Immune Thrombocytopenia in Children / 中国实验血液学杂志

OBJECTIVE@#To explore the influencing factors in children with chronicity immune thrombocytopenia (ITP), and to provide basis for judging the prognosis and treatment in children with ITP.@*METHODS@#The clinical data of children with ITP admitted to The Second Affiliated Hospital of Anhui Medical University in the past 5 years were retrospectively analyzed and followed up for more than 1 year. According to the inclusion criteria, the eligible cases (328 cases in total) were selected and collected through medical record system retrieval, outpatient clinic and telephone follow-up. Independent influencing factors affecting the prognosis of children with ITP were obtained through single-factor and multi-factor logistic analysis, and their predictive value for the prognosis of ITP in children were evaluated.@*RESULTS@#Of 328 children with ITP, 208 were newly diagnosed with ITP (64%), 54 were persistent ITP (16%), 66 were chronic ITP (20%), and the remission rate within 1 year was 79.9%. The results of univariate analysis showed that, age, pre-morbidity history of infection and vaccination, antinuclear antibodies, initial absolute lymphocyte count（ALC） and treatment options were related to the prognosis of the children (P<0.05). Multivariate analysis showed that the history of infection and vaccination before onset, initial treatment options, and ALC at the time of initial diagnosis were independent factors affecting the prognosis of children with ITP (P<0.05). The time for platelet recovery to 100×10@*CONCLUSION@#The initial treatment plan combined with IVIG can reduce the occurrence of chronicity in children with ITP, and its efficacy is better than that of the single corticosteroids group (the platelet recovery time is shorter); history of preceding infection or vaccination, ALC at the time of initial diagnosis are independent factors affecting the prognosis of children with ITP, and the combination of the two shows a better predictive value for the prognosis.

Relationship between Nutritional Status and Nosocomial Infection in Children with Acute Lymphoblastic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To study the risk factors and infection characteristics of nosocomial infection in children with acute lymphoblastic leukemia (ALL) and analyze the relationship between different nutritional status and nosocomial infection, early treatment response.@*METHOD@#The clinical data of 133 children with ALL treated with CCCG-ALL-2015 from June 2016 to June 2019 (chemotherapy stage, risk level, MRD), infection during hospitalization (course of infection, laboratory indicators, sites of infection, outcome) and nutritional status (sex, age, height/ length, weight) were enrolled. The Chi 2 test and Logistic regression analysis were used for statistical analysis.@*RESULTS@#The rate of nosocomial infection was 19.9% in 133 children with ALL, in which 3 were infection-related death. Sex, immunophenotype and risk showed no significantly affect on the occurrence of nosocomial infection (P＞0.05), but neutrophil count, hemoglobin level, platelet count, chemotherapy stage, length of stay in hospital and nutritional status showed affect on the occurrence of nosocomial infection (P＜0.05). Logistic multivariate regression analysis showed that chemotherapy stage, length of hospital stay, neutrophils and nutritional status were the independent risk factors, in which the respiratory tract infection was the most common. Gram-positive bacteria, Gram-negative bacteria and fungi accounted for 44.1%, 52.9% and 2.9% respectively. The negative rate of MRD in day 19 and day 46 between different nutritional status groups showed statistically significant (P＜0.05).@*CONCLUSION@#Neutrophil count, chemotherapy stage, length of stay in hospital and nutritional status are independent risk factors for nosocomial infection. Among of them, nutritional status negatively correlated with nosocomial infection, and the poorer nutritional status, the higher risk of nosocomial infection. Malnutrition, overweight and obesity can affect the early treatment response of ALL children. The level of nutrition at first diagnosis can be used as a bad factor to evaluate the early treatment response of ALL children.

Clinical Analysis of Deep Venous Thromboembolism in Children with Acute Lymphocytic Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore clinical characteristics and outcome of deep vein thrombosis(DVT) in children with acute lymphoblastic leukemia (ALL).</p><p><b>METHODS</b>A tatol of 266 patients were diagnosed as ALL from January 1, 2010 to May 31, 2016. The clinical data of 12 cases of patients with DVT were retrospectively analyzed, 183 cases diagnosed before January 1, 2015 were received chemotherapy with the scheme of SCMC-05. The other cases were treated by the scheme of CCCG. All the patients received central venous catheter.</p><p><b>RESULTS</b>The DVT happened in 12 cases including 10 cases of limb DVT and 2 cases of intacranial venous sinus thrombosis. The DVT mostly occured in intermediate risk ALL patients, the infection and coagulopathy existed in most patients. They were treated with low molecular heparin(LWHP), among them 5 cases were given extubation; the thrombus disappeared in 6 cases after 1 week; the intracranial venous sinus thrombosis in 1 case did not obviously improved after 6 months of treatment. The ALL children with DVT were treated with LWHP when using L-ASP, as a result no thrombuses happened.</p><p><b>CONCLUSION</b>Centralvenous catheter and chemotherapeutic drugs were the major cause of DVT. Abnormal coagulation, infection, and risk stratification are another risk factors for thrombosis. ALL children thrombosis are benefited from LWHP prevention when using L-ASP again.</p>

Correlation of SOCS3 mRNA Expression Level with Clinical Characteristics and Early Response to Treatment in Pediatric Acute Lymphoblastic Leukemia / 中国实验血液学杂志

<p><b></b>Obejective: To investigate the expression level of suppressor of cytokine signaling SOCS3 mRNA in children's acute lymphoblastic leukemia(ALL); to analyze the relationship between the expresion level of SOCS3 mRNA and disease status and risks of ALL, and to explore the application of SOCS3 mRNA in evaluation of ALL disease status, risk and target therapy.</p><p><b>METHODS</b>The expression levels of SOCS3 mRNA in bone marrow mononuclear cells from 45 cases of newly diagnosed ALL at initial diganosis and induction remission and 13 normal children as controls were detected by SYBR Green fluorescence quantitative PCR; the correlation of SOCS3 mRNA expression level with risk and clinical characteristics of ALL was analyzed by means of statistical method; the immunofluorescence histochemistry method and laser confocal microscopy were used to detect the sites and expression level of SOCS3 mRNA in bone marrow samples of ALL patients.</p><p><b>RESULTS</b>The SOCS3 mRNA expression level at initial diagnosis of 45 ALL patients was significantly lower than that of normal controls (P<0.05), and that in induction remission of 45 patients was not significant different from normal controls (P<0.05); the SOCS3 mRNA expression level at initial diagnosis of patients with standasd and high risk was higher than that of patients with low risk (P<0.05). The 45 patients were divided into high expression and low expression groups according to SOCS3 mRNA expression level at initial diagnosis by median method, comparison of clinical characteristics in 2 groups was found that the SOCS3 mRNA high expression group had even more higher leukocyte count in peripheral blood, even more higher LDH level and much more poor prognostic genes; in addition, the high expression group showed more higher risk in comparison between 2 group.</p><p><b>CONCLUSION</b>The SOCS3 mRNA expression is down-regulated at initial diagnosis, while recovered to normal level after induction remission of disease, thus the SOCS3 can be used as an indicator for evaluation of disease status. The high expression of SOCS3 mRNA up-regulates the disease risk, therefore the SOCS3 mRNA can be used as a factor for evaluation of ALL risk. The treatment of ALL via regulation of SOCS3 mRNA expression level maybe can become a new way. The regulation of SOCS3 mRNA expression level maybe can become a new way for treatment of ALL.</p>

Clinical Significance of Hepcidin in the Diagnosis of Infant Iron Deficiency Anemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical diagnostic value and significance of hepciden level by detecting the expression of serum hepcidin before and after treatment of infant iron deficiency anemia (IDA) with or without vitamin D deficiency.</p><p><b>METHODS</b>A total of 60 cases of infamt IDA were divided into A and B groups, the group A consisted of 20 IDA infants with vitamin D deficiency, group B consisted of 48 IDA infants without vitamin D deficiency and the control group included 26 healthy infants. Blood examination including HGB, MCV, MCH and MCHC was performed by hematological analyzer, the level of serum ferritin was assayed by chemiluminescence immunoassay, the levels of hepcidin and 25- (OH) D were assayed by ELISA.</p><p><b>RESULTS</b>The levels of serum hepcidin in group A, B and control group before treatment were (29.16 ± 7.50), (27.11 ± 7.10) and (29.25 ± 8.39) ng/ml, respectively (P > 0.05). The level of serum hepcidin in group A and B after treatments was significantly higher than that in control group [ (36.21 ± 5.68) ng/ml vs (29.25 ± 8.39) ng/ml, P < 0.01; (34.16 ± 4.54) ng/ml vs (29.25 ± 8.39) ng/ml, P < 0.01]; but there were no significantly difference between group A and B (P > 0.05). The serum ferritin positively correlated with hepcidin in group B both before and after treatments (r = 0.352 and 0.367, P < 0.05, P < 0.05).</p><p><b>CONCLUSION</b>The level of serum hepcidin has an important significance in poccess of evaluatng for therapeutic effect in infant iron deficiency anemia, but the interference effect of vitamin D deficience should be eliminated when the expression level of hepcidin is applicated for diagnosis and differential diagnosis.</p>

The effect of parathyroid hormone on osteogenic activities of human osteosarcoma cell line SaOS-2 / 中国地方病学杂志

Objective To observe the effects of recombinant human parathyroid hormone 1 to 34(referred to as hPTH) on the expression level of alkaline phosphatase(ALP) and bone gla protein(BCP) in human osteosarcoma cell line SaOS-2(referred to as SaOS-2 cells). Methods SaOS-2 cells were subcultured and treated with 1, 10 and 100 nmol/L hPTH for 12, 24 and 48 h. Total cellular RNA was extracted, cDNA was synthesized by reverse doses of hPTH, different duration of action, and their interaction on the expression level of ALP mRNA of SaOS-2 cells was significantly different(F = 29.32, 2.92, 7.64, all P < 0.05). The expression level of ALP mRNA(0.78 ± 0.43, 0.71 ± 0.05, 0.75 ± 0.19, 0.76 ± 0.14) of SaOS-2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h(1.01 ± 0.16, 1.37 ± 0.38, 1.49 ± 0.16, 2.52 ± 0.70, all P< 0.05) and 24 h (1.80 ± 0.47, 1.30 ± 0.36, 1.27 ± 0.17, 1.17 ± 0.11, all P< 0.05). The expression level of ALP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 hours was higher than that of the control(P < 0.05); the expression level of ALP mRNA of SaOS-2 cells after treatment with 1, 10 and 100 nmol/L hPTH for 24 h interaction on the expression level of BGP mRNA of SaOS-2 were significantly different (F = 8.26, 10.33, 5.51, all P< 0.05). The expression level of BGP mRNA(1.17 ± 0.28, 0.98 ± 0.08, 0.92 ± 0.17 and 0.84 ± 0.59) of SaOS2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h( 1.01 ± 0.14, 1.21 ± 0.18, 1.34 ± 0.30, 1.68 ± 0.62, all P< 0.05), and 24 h(1.71 ± 0.35, 1.41 ± 0.47, 1.28 ± 0.31 and 1.01 ± 0.18, all P < 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 h was higher than that of those groups treated with 0 and 1 nmol/L hPTH(all P< 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 10 and 100 nmol/L hPTH for 24 h and 48 h was lower than those of the control(all P < 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 24 hours was lower than that the group treated with 1 nmol/L hPTH(P < 0.05). Conclusions In vitro, hPTH significantly enhances osteogenic activities of human osteoblast in a short time, however, with prolonged stimulation time, osteogenic activity can show a downward trend.

Effect of different doses of recombinant human bone morphogenetic protein 2 on osteogenic activities of human osteosarcoma cell line SaOS-2 / 中国地方病学杂志

Objective To investigate the effect of recombinant human bone morphogenetic protein 2 (rhBMP-2) on the osteogenic activities of human osteosarcoma cell line SaOS-2. Methods SaOS-2 cells were exposed to rhBMP-2 for 12,24,48 h at 0(control) ,2,20,200 μg/L, respectively. The mRNA expression of alkaline phosphatase(ALP) and bone gla(BCP) were detected by real time polymerase chain reaction. Results The mRNA expression of ALP and BGP of SaOS-2 cells increased gradually with rhBMP-2. The mRNA expression of ALP of the 20 μg/L group exposed for 48 h(1.60 ± 0.64), and the 200 μg/L group exposed for 12,48 h(1.70 ± 0.41, 1.80±0.19) were significantly higher than those of control (12 h: 0.80±0.25, 48 h: 0.74±0.21, allP<0.05). The mRNA expression of BGP of the 2 μg/L group exposed for 24 h(1.67 ± 0.33), the 20 μg/L group exposed for 12,24 h(2.42 ± 0.13,1.82 ± 0.14) and the 200 μg/L group exposed for 12,24 h(1.46 ± 0.11,1.24 ± 0.07) were significantly higher than those of control( 12 h: 1.01 ± 0.14, 24 h: 0.84 ± 0.12, all P< 0.05). Conclusions rhBMP-2 can promote the mRNA expression of ALP and BGP of SaOS-2 cells. They have a dose-response relationship, but represent a different dose-response effect.

Effect of fluoride on bone metabolism in rats / 中国地方病学杂志

Objective To study the effect of fluoride on bone metabolism in rats, and to understand the mechanism of pathogenesis of skeletal fluorosis. Methods A total of 80 Wistar rats were randomly divided into 4 groups that included control group (distilled water), low-dose group(NaF, 50 mg/L), medium-dose group (NaF,100 mg/L) and high-dose group(NaF, 150 mg/L), respectively. After being bred for 12 weeks, the rats were put to death (etherization). Incidence of dental fluorosis was estimated, and serum was collected. Radioimmunoassay was employed to detect the levels of osteocalcin (BGP), parathyroid hormone (PTH) and calcitonin (CT), respectively.Colorimetry method was employed to determine the levels of alkaline phosphatase (ALP) and acid phosphatase (ACP). Results Incidence of dental fluorosis between the four groups was significantly different statistically(x2 =82.81 ,P ＜ 0.01 ). The incidence was significantly different(x2 = 22.67, 40.00, 40.00, all P＜ 0.01 ) between low-dose ( 80%, 16/20), medium-dose ( 100%, 20/20), high-dose groups ( 100%, 20/20) and control group (0,0/20),respectively. Serum levels of BGP, PTH, CT were significantly different between the groups(F = 38.614, 20.778,3.023, P ＜ 0.01 or ＜ 0.05). There was no significant difference between the four.groups of ALP and ACP in serum (F = 0.609,2.895, all P ＞ 0.05 ). Serum BGP in low-dose, medium-dose and high-dose groups[ ( 19.60 ± 12.79),(33.41 ± 10.81 ), (39.46 ± 9.51 )mg/L, respectively] was significantly higher than that of the control group[ (7.35 ± 3.22)mg/L, all P ＜ 0.01 ]. Serum PTH in low-dose, medium-dose and high-dose groups[ (72.27 ± 25.38), (67.80 ± 12.01), (106.52 ± 36.37)pmol/L] was significantly higher than that of the control group[(47.08 ± 9.22)pmol/L,all P ＜ 0.01 ]. Serum PTH of the high-dose group was significantly higher than that of the low-dose and the mediumdose groups(all P ＜ 0.01 ). Serum CT in medium-dose and high-dose groups[ ( 13.39 ± 2.07), ( 15.05 ± 4.77)pmol/L ] was significantly lower than that of the control group[ (26.06 ± 28.31 ) pmol/L, all P ＜ 0.05 ] and also significantly lower than that of the low-dose group [ (24.49 ± 14. 10) pmol/L, all P ＜ 0.05 ]. Conclusions Fluoride affects bone metabolism in rats, BGP, PTH and CT play a key role in the pathogenesis of skeletal fluorosis.