Objective To prepare an internal quality control substance of free hemoglobin (FHb),and evaluate its perform-ance.Methods ① 5 normal whole human blood samples with known hemoglobin (Hb)concentration was selected and dilu-ted with distilled water in the ratio of 1∶500,1∶1 000,1∶2 000,1∶4 000 respectively.After 20 FHb solutions was fully mixed,the concentration of FHb solutions was tested and compared with its theoretical data.②One of the highest value and the lowest value solutions were chosen as qualitye control substance,packed and stored in -20℃ fridge.Each group was an-alyzed 20 days continuously in the routine conditions,where average (-x )of the groups,standards deviation (s )and coeffi-cients of variation (CV)were calculated and Levey-Jennings control chart was conducted.Then,the test was repeated every week,and the data on the chart was updated accordingly until (CV)of the reagents were changed.The average (-x)of the-groups,standards deviation (s)and coefficients of variation uality control material was calculated during the whole test peri-od.Results ①The theoretical value (-x ±s)of the concentration of free hemoglobin was 125.44±93.04 mg/L,and the actu-al value was 125.22±93.08 mg/L,there was no statistic significance (t value was 0.706,P >0.05).② During 20 continu-ously daystest,the detected values (-x ±s)of the highest quality control material was 303.55±3.70 mg/L;the values of the lowest quality control material was 69.29 ± 1.88 mg/L.The coefficients were 1.22% and 2.68% respectively.Both the highest and the lowest quality control material were under control during the whole detecting period.The (-x ±s )values of the highest was 302.56±3.99 mg/L,the CV value of it was 1.32%;The (-x ±s)values of the lowest was 69.04±1.88 mg/L,the CV value of it was 2.71%.Conclusion The preparation method of internal quality control substance of free hemoglo-bin is convenient and stable,and was applicable in blood station’s and clinical laboratory.

Objective:To analyze the clinical characteristics and diagnosis and treatment of children with Kasabach-Merritt phenomenon (KMP).Methods:A retrospective analysis was conducted on the clinical data and follow-up data of 8 patients diagnosed KMP in Beijing Children′s Hospital, Capital Medical University from January 2016 to January 2019.The clinical data included laboratory examination, diagnosis, treatment and prognosis.Results:Among the 8 children with KMP, 6 cases were male and 2 cases were female.The median onset age was 4 (0-17) months, 2 cases of neonatal onset.The median onset to the diagnosis time was 59 (34-140) days; 6 cases with bone destruction; 6 cases had misdiagnosis and mistreatment history, they were misdiagnosed as idiopathic thrombocytopenic purpura, Evans syndrome, abnormal bone and joint development; 4 cases were Kaposiform hemangioendothelioma; 8 cases were used alone or combined with the application of hormones, Sirolimus, and Vincristine, 7 patients underwent interventional therapy.All patients survived with a median follow-up period of 487 (112-1 033) days.Median time of platelet count returned to normal was 24.5 (7-60) days, and median time of fibrinogen returned to normal was 20 (7-30) days.Median time of D-dimer dropped to a normal was 105 (40-240) days.Conclusions:Children with concurrent platelet count and coagulation abnormalities should be considered with KMP.Doctors need to identify the potential visceral vascular lesions.Early diagnosis and treatment are important, which can improve the clinical prognosis of patients.

The clinical and laboratory data of a patient with chronic refractory immune thrombocytopenia (ITP) who had a significant increase in the proportion of inhibitory T cells in the hematological oncology center of Beijing Children′s Hospital Affiliated to Capital Medical University in February 2018 and regularly followed up in the outpatient department received a retrospective analysis.This 8-year-old patient′s clinical features were 6 years of skin and mucous membrane bleeding spots and petechiae recurring, with occasionally nasal epistaxis.Physical examination: cardiopulmonary abdomen and nervous system examinations are normal, and no superficial lymphadenopathy is touched.Blood routine indicated that the platelets were 2.00×10 9/L, and white blood cell count and hemoglobin level were normal.Bone marrow suggested that hyperplasia was significantly active, with more than 300 megakaryocytes.The patient was diagnosed with ITP, and he was treated with first-line treatment with gamma globulin and oral Corticosteroids.The first-line treatment with high-dose Dexamethasone therapy was repeated, and the second-line treatment was low-dose Rituximab combined with high-dose Dexamethasone.Evaluations had been conducted with every relapsed course, and Cyclosporine was administered orally on the basis of the highly suppressive T cells.After half a month, the child achieved partial remission and continued for 3 months, and then reached a complete remission of 6 months (till this paper). It is suggested that, with ITP as an immunological disease with high heterogeneity, the immune abnormality index is expected to become a breakthrough in the development of precise treatment.

We studied the epidemiological characteristics of human bocavirus 4 (HBoV4) in children with a- cute gastroenteritis in Lanzhou (China). A total of 331 stool specimens were collected from children aged < 5 years with acute diarrhea at the First Hospital of Lanzhou University between July 2012 and June 2013. Specimens of HBoV were identified by nested polymerase chain reaction assays. Compared with related sequences in GenBank, the HBoV-positive strain isolated in the present study was,quite surprisingly, a rare genotype named HBoV4. This strain was a typical HBoV4,with high levels of nucleotide and amino acid homology to the Thailand strain, JQ267789 (98.9% and 98.7%, respectively), and the USA strain, GQ506568 (97.6% and 97.4%, respectively). This is the first report of HBoV4 as the causative agent for acute gastroenteritis in pediatric patients in China. This strain is one of two genotypes of HBoV that are currently circulating.
Child, Preschool ; China ; Feces ; virology ; Female ; Gastroenteritis ; virology ; Human bocavirus ; classification ; genetics ; isolation & purification ; Humans ; Infant ; Male ; Molecular Sequence Data ; Parvoviridae Infections ; virology ; Phylogeny

A method for simultaneous determination of 12 kinds of chlorinated disinfection byproducts (DBPs) in drinking water was developed based on liquid-liquid extraction gas chromatography equipped with electron capture detector (GC/ECD).The procedural standard calibration was adopted to eliminate the interference of different matrix.The method detection limits for 12 DBPs were 0.08-0.21 μg/L and the entire analytical procedure was finished in 21.50 min.The recoveries were in the range of 80.9%-115.7% and the relative standard deviations (RSD) were between 0.9% and 9.9% at different concentration levels (5 and 50 μg/L) in tap water and surface water.The correlation coefficients for all 12 kinds of DBPs were greater than 0.99 in the linearity range of 0.5-200 μg/L.The method was applied to determine DBPs in drinking water and source water.This method was rapid and competent for detection of volatile DBPs in drinking water.

The dosage-efficacy/toxicity relationship of the 50% alcohol extracts of Polygonum multiflorum was comparatively investigated on either normal or CCl4-induced chronic liver injury rats, by determining the general condition, serum biochemical indices and liver histopathology, coupled with the factor analysis. The dosages were 10 and 20 g raw materials per kg body weight. Compared with the normal control group, the normal high dose group showed significant increases of the serum alanine transaminase (ALT), total bilirubin (TBIL), high mobility group box 1 (HMGB-1) and interleukin-1β (IL-1β) (P < 0.05 or P < 0.01), as well the frequent incidences of inflammatory cell infiltration, hepatic sinus enlargement and fiber stripes formation in histopathological sections. Compared with the model control group, the model low dose group showed significant declines of serum ALT, aspartate transaminase (AST) and total bile acid (TBA) (P < 0.05), as well the alleviation of vacuoles of hepatocytes, but no amelioration of the inflammatory cell infiltration and fibrous tissue hyperplasia; moreover, the model high dose group showed significant degeneration declines of serum HMGB-1, tumor necrosis factor-α (TNF-α) and IL-1β (P < 0.05, P < 0.01), as well the evident alleviation of vacuoles degeneration of hepatocytes, inflammatory cells infiltration and fibrosis degree. The factor analysis showed that the low dosage treatment had almost neither injuring effect on the normal rats nor protective effect on the model rats; while the high dosage treatment showed observable injuring effect on the normal rats, expressed by the significant increases of the factor-1 (HMGB-1, TNF-α and IL-1β as the main contributors) and factor-2 (TBIL, ALT and TBA as the main contributors) relative to the normal control group. The liver protective effect of the high dosage treatment could be observed with the significant reduction of the factor-1, indicating the effective alleviation of the expression of inflammatory cytokines. In conclusion, it could illustrated the phenomenon of symptom-based prescription theory of Polygonum multiflorum on rat livers: the high dosage of the herb had either an injuring effect on normal rats, or a therapeutic effect on the rats with chronic liver injury.

Objective To study the main genes molecular characteristics and evolutionary relationship of porcine and human group A rotavirus (GARV) type G9 in Hebei Lulong region.Methods A stool specimen from diarrhea piglet marked as LLP48 in 2008 in Hebei Lulong region,which was G9 GARV positive and 4 stool specimens collected from hospitalized children with diarrhea under the age of five from 2009 to 2011 in Lulong region which were type G9 GARV positive were selected and the main genes were amplified by RT-PCR.After sequencing,the obtained genes were analyzed through MEGA,DNAStar software and other biological sequence alignment,homology and phylogenetic analyses were performed.Results Porcine GARV LLP48 in Lulong region and four strains of Lulong human GARV were highly homologous in VP7,VP6,NSP4 and NSP2 gene fragments,and nucleotide and amino acid homology respectively was 89.4％-94％ and 94.8％-98.2％ ; VP4 fragment of the nucleotide homology (amino acid) is low,71.4％-71.6％ (68.2％-69.0％).Phylogenetic analysis showed that LLP48 genes encoding VP7,VP6,NSP4 and NSP2 were closely related to the strain derived from human source,and VP4 gene closely related to the strains derived from porcine source.Conclusion Lulong porcine GARV LLP48 strain may be a natural reassorted product of porcine VP4 and human VP7,VP6,NSP4 and NSP2.

The liver injury induced by Polygonum multiflorum Thunb. (PM) was investigated based on idiosyncratic hepatotoxicity model co-treated with lipopolysaccharide (LPS) at a non-hepatotoxic dose. Sprague-Dawley (SD) rats were intragastrically administered with three doses (18.9, 37.8, 75.6 g crude drug per kg body weight) of 50% alcohol extracts of PM alone or co-treated with non-toxic dose of LPS (2.8 mg·kg(-1)) via tail vein injection. The plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were assayed and the isolated livers were evaluated for histopathological changes. The dose-toxicity relationships of single treatment of PM or co-treatment of LPS were investigated comparatively to elucidate the idiosyncratic hepatotoxicity of PM. The results showed that no significant alterations of plasma ALT and AST activities were observed in the groups of solo-administration of LPS (2.8 mg·kg(-1), i.v.) or different dosage (18.9, 37.8 and 75.6 g·kg(-1), i.g.) of PM, compared to normal control group (P > 0.05); while significant elevations were observed in the co-administration groups of PM and LPS. Treatment with LPS alone caused slight infiltration of inflammatory cells in portal area but no evident hepatocytes injury. Co-treatment with LPS and PM (75.6 g·kg(-1), i.g.) caused hepatocyte focal necrosis, loss of central vein intima and a large number of inflammatory cell infiltration in portal areas. When further reduce the dosage of PM, significant increases of plasma ALT and AST activities (P < 0.05) were still observed in co-administration groups of LPS and PM (1.08 or 2.16 g·kg(-1)), but not in LPS or PM solo-administration groups. Nevertheless, the co-treatment of low dosage of PM (0.54 g·kg(-1)) with LPS did not induce any alteration of plasma ALT and AST. In conclusion, intragastric administration with 75.6 g·kg(-1) of PM did not induce liver injury in normal rats model; while the 2 folds of clinical equivalent dose of PM (1.08 g·kg(-1)) could result in liver injury in the LPS-based idiosyncratic hepatotoxicity model, which could be used to evaluate the idiosyncratic hepatotoxicity of PM.

Objective: To compare the efficacy and safety of short course and high-dose Dexamethasone (HDD) and conventional Prednisone as first-line strategy for children newly diagnosed as primary immune thrombocytopenia (ITP). Methods: This study analyzed pre-experimental data of a single center prospective randomized controlled clinical trial.Newly diagnosed but untreated ITP patients enrolled at the Department of Blood and Cancer Center, Beijing Children's Hospital, Capital Medical University from November 2016 to May 2017 were randomized into HDD group[Dexamethasone 0.6 mg/(kg·d), intravenous injection for 4 days]and Prednisone group[Prednisone 2 mg/(kg·d) for 14-28 days and then tapered within 1-2 months, the course of treatment less than 3 months]. Initial response, sustained response and adverse effects after therapy were observed in 2 groups. Results: Sixty-six children with ITP were included in the study: 32 patients were in the HDD group and 34 patients were in the Prednisone group.Two groups were matched in the baseline characteristics including gender, age, platelet counts and disease course before therapy and bleeding assessment (all P>0.05). The initial response (the response of HDD group within 10 days of treatment and Prednisone group within 28 days of treatment): overall initial response had no statistical difference between the HDD group and the Prednisone group[90.6%(29/32 cases) vs.100.0%(34/34 cases), χ²=1.528, P>0.05]. HDD group had a lower incidence of complete response compared with that in the Prednisone group[54.4%(19/32 cases) vs.94.1%(32/34 cases), χ²=11.330, P=0.001]; median time of response in two groups showed no statistically difference (2 d vs.1 d, Z=-0.149, P>0.05). There was no significant difference in the recovery of skin and mucosal bleeding after treatment between the Dexamethasone group and the Prednisone group (Z=-1.413, -1.031, all P>0.05). The sustained response (the response lasted for up to 6 months and above): overall and complete sustained response had no statistically difference between the HDD group and the Prednisone group [92.9%(26/28 cases) vs.85.3%(29/34 cases), P=0.594; 78.9% (15/19 cases) vs.81.3%(26/32 cases), P=1.000]. Log-rank test showed no significant difference in the duration of response between 2 groups (P=0.341). The side effects in the Prednisone group included weight gain or Cushing sign (94.1%) and mental and emotional changes (23.5%); in the HDD group 15.6% of children had infection, without other glucocorticoid-related side effects.There was no significant difference in the incidence of infection between two groups[15.6%(5/32 cases) vs.26.5%(9/34 cases), P=0.281]. All of the above infections were of respiratory tract infections and mild gastrointestinal infections. Conclusions Efficacy of the HDD group in the initial and sustained responses is similar, but side effects were apparently lower compared with that in the Prednisone group.However, a large multicenter randomized controlled clinical study is necessary to confirm this result.

Objective To explore the clinical features and points of diagnosis and treatment for congenital thrombotic thrombocytopenic purpura (TTP) in children.Methods The clinical manifestations,laboratory tests,genetic analysis and treatments of 5 children with congenital TIP hospitalized in Beijing Children's Hospital,Capital Medical University from February 2015 to July 2017 were analyzed retrospectively.Results Among the 5 children with congenital TTP diagnosed by genetic monitoring and enzymology,there were 1 male and 4 females,3 cases had suspicious positive family history,the age of onset was several hours after birth (range several hours after birth to 28 months).The main clinical manifestations were recurrent moderate to severe thrombocytopenia in 5 cases,mild to moderate hemolytic anemia in 4 cases,proteinuria or hematuria in 2 cases,and nervous system involvement in 1 case.The recurrence time was 1.5 (range 1.0 to 5.0) times per year and most of the inducing factors were respiratory and (or) digestive tract infections.Laboratory test showed that ADMATS13 enzyme activity were 0 in 4 cases,the enzyme activity was 100％ in 1 case due to plasma infusion before examination.ADMATS13 enzyme antibody detection of all 5 cases were negative.Genetic analysis of all 5 children showed complex heterozygous mutations at different loci of ADAMTS13 gene,among which 8 loci were previously unreported,details are as follows:missense mutations in 4 cases (c.1564T＞C(p.522C＞R),c.1510G＞T(p.504D＞Y),c.4154A＞C (p.1385Q＞P) and c.G3854C (P.R1285P));frameshifi mutations in 3 cases(c.2875_2876insT (p.959Lfs29),c.2362_2363delGG (p.788G＞Gfs56) and c.1335delC (p.F445fs)),shear mutation in one case(IVS21 + 1A＞G).The patients in the acute phase were all treated with fresh frozen plasma infusion (10 ml/(kg· d)),continuous application for 7-14 days).Platelets gradually returned to normal and clinical symptoms improved.The follow-up time was 27 months (range 11-35 months).All the children survived,among whom 2 cases were treated with prophylaxis and monitoring platelet stability above 200×109/L,3 cases were treated on-demand only when platelet decreased and monitoring platelet stability above 100× 109/L.Conclusions The main clinical manifestation of congenital TTP is recurrent thrombocytopenia with or without hemolytic anemia.The key point of treatment is plasma infusion.Genetic testing is helpful for early diagnosis.