BACKGROUND:It is important to establish an ideal mouse model of severe aplastic anemia for investigating the mechanism and finding new therapies for aplastic anemia. OBJECTIVE:To establish a severe aplastic anemia mouse model by using recombinant human interferon-γand busulfan. METHODS:Sixty healthy Kunming female mice were randomly divided into two groups:model group (n=50) and control group (n=10). The model group was given recombinant human interferon-γat a dose of 1×104 U/d by intraperitoneal injection and busulfan at a dose of 18 mg/(kg·d) through stomach feeding for 7 days. The same volume of physiological saline was given to control group. Multi-parameters, including general condition, body weight, blood cellcount, morphology and biopsy of bone marrow were analyzed in two groups. RESULTS AND CONCLUSION:At day 7 after treatment, the weight, white blood cellcount, hemoglobin, blood platelet, reticulocyte count in model group were significantly lower than control group (P<0.05). Bone marrow smears and biopsy of model group showed marked reduction of bone marrow proliferation and increases of percentages of non-hematopoietic cellclusters and adipose tissue. The oil drop and fat vacuole were apparently seen in the model group. Severe aplastic anemia mouse model can be established by using recombinant human interferon-γand busulfan successful y, which is economic, stable and easy to operate.

Objective To observe the effects of endoplasmic reticulum stress (ERS) on the activation of monocytes induced by high glucose and explore the underlying mechanism.Methods The monocyte cell line THP-1 was stimulated with high glucose,and then treated with molecular chaperone betaine.The levels of glucose regulation protein 78 (GRP78) and p-JNK,which were associated with ERS were detected by real-time PCR and Western blotting.The proliferation of the cell line was detected by MTT method.Transwell and immunofluorescence were applied to observe the chemotaxis and phenotype of cells respectively.Results The levels of GRP78 and p-JNK of THP-1 cells stimulated by high glucose were significantly increased compared with the normal control group (all P ＜ 0.05).The proliferation and chemotactic were also enhanced (all P ＜ 0.05).The number of cells in M1 phenotype was increased remarkably (P ＜ 0.05).All the indexes above could be rescued by betaine.Conclusion The activation of THP-1 cells can be induced by high glucose through ERS,while molecular chaperone betaine can reverse the activation.

Objective To study the mechanism of restenosis following percutaneous transluminal coronary angioplasty(PTCA),and to replicate a dynamic model of cell proliferation and remoulding of vascular wall at different time points in rabbits after intimal injury.Methods The model of restenosis in common carotid artery was established by balloon injury in 70 rabbits.The indexes such as lumen area,thickness and area of intima and media,and cross sectional area bounded by the external elastic lamina(EELA) were respectively measured by computer image analysis technology at the 1st,3rd,5th, 7th,14th,28th and 35th day after the injury.Results Endothelial cells were denudated at the 1st day after injury.The proliferation of vascular smooth muscle cell(VSMC) was detected on the surface of lumen at 3 days after injury.At the 7th day after injury,the neointima was formed and continuously thicken.The thickness and area of the neointima as well as extracellular matrix were gradually increased after 14 days,and were maximal after 35 days.The thickness and area of media were also gradually increased during 3～14 days and decreased after 28 days.Compared with non-injured vessel,the medial area was obviously increased at the 14th day.The lumen area was decreased at the 5th～7th day after injury and was obviously less than that of non~injured vessel after 14 days.The EELA was gradually increased at the 1st～7th day after injury,and reached its maximum at the 14th day.The EELA was declined gradually after 28 days.Conclusion The progress of restenosis(RS) can be simulated through the model of restenosis in common carotid artery of rabbit established by balloon injury.The intimal proliferation and vascular remodeling are the leading pathogenesis of restenosis.

OBJECTIVETo identify the changes in serum insulin like growth factor-I (IGF-I) and IGF binding proteins (IGFBPs) in children with nephrotic syndrome (NS) and the effect of glucocorticoid on serum IGF-I and IGFBPs.

METHODSWe measured serum IGF-I and IGFBPs levels by radioimmune assay and immune radiomagnetic assay in 36 children with NS, consisting of an active stage group (ANS, n = 12), a remission stage group (RE, n = 12), an active stage group with glucocorticoid treatment (GNS, n = 12), and a normal control group (NC, n = 10).

RESULTS1) Compared to NC, serum levels of IGF-I and IGFBP-3 were decreased (P < 0.01); serum levels of IGFBP-1 and IGFBP-2 were increased (P < 0.01) in the ANS group. 2) Serum levels of IGF-I and IGFBP-3 were higher and IGFBP-1 and IGFBP-2 were lower in the RE Group than in theANS Group (P < 0.01). 3) Compared to the ANS group, serum levels of IGF-I and IGFBP-3 were increased (P < 0.01) and serum levels of IGFBP-1 and IGFBP-2 were decreased (P < 0.01) in the GNS group. 4) A correlation was found between serum levels of IGFBP-3 and albumin in the active stage group (r = 0.76, P < 0.01). There was also a correlation between serum levels of IGF-I and IGFBP-3 and an inverse correlation between the serum level of IGF-I and serum levels of IGFBP-1 and IGFBP-2 in the ANS group. No other correlations were observed.

CONCLUSIONSThe serum levels of IGF-I and IGFBPs are altered in children in the active stage of NS, but return to normal in the remission stage. GC treatment may influence serum IGF-I and IGFBPs in children with NS. Changes in IGF-I and IGFBPs levels may play a role in the growth retardation of NS children.

Child ; Dexamethasone ; pharmacology ; Female ; Glucocorticoids ; pharmacology ; Humans ; Insulin-Like Growth Factor Binding Proteins ; blood ; Insulin-Like Growth Factor I ; analysis ; Male ; Nephrotic Syndrome ; blood

BACKGROUND:As the high proliferation and low apoptosis of the bone marrow in polycythemia vera patients, hematopoietic stem cels transplanted into NOD/SCID mice can differentiate into erythroid cels, but whether hematopoietic stem cels transplantation could improve the hematopoietic function of aplastic anemia mice is not yet reported. OBJECTIVE:To investigate whether transplantation of bone marrow mononuclear cels with JAK2V617F mutation from polycythemia vera patients can influence hematopoietic reconstruction in aplastic anemia mice. METHODS:Severe aplastic anemia mouse models were established by using recombinant human interferon-γplus busulfan, and then, these mouse models were randomly divided into experimental group (n=10) and control group (n=10). Bone marrow mononuclear cels isolated from polycythemia vera patients with positive JAK2V617F mutation were transplanted into the mice in the experimental group via tail vein at 5 days after drug withdrawal.The same volume of normal saline was administered to the control group. Routine peripheral blood test, morphology of bone marrow cels, bone marrow biopsy, and percentage of CD45+ cels in the peripheral blood and marrow were determined at 14 days after transplantation. RESULTS AND CONCLUSION: At 14 days after transplantation, pancytopenia occurred in the experimental group, bone marrow smears showed scattered lymphocytes and hematopoietic progenitors, and bone marrow biopsy presented that hematopoietic tissues were reduced and a smal amount of granulocyte cels and erythroblasts could be seen, but megakaryocytes were rare. In contrast to the control group, there was no improvement in the hematopoietic function of mice in the experimental group. CD45+ cels were detectable in the peripheral blood and bone marrow in the experimental group, but not in the control group; and a higher percentage of CD45+ cels was measured in the bone marrow than in the peripheral blood of experimental group mice. Experimental findings indicate that bone marrow mononuclear cels from polycythemia vera patients with positive JAK2V617F mutation can be engrafted into aplastic anemia mice, but cannot improve the hematopoietic function of mice.

OBJECTIVETo study the levels and roles of cytokines TNF-α, IL-6 and IL-10 in bronchoalveolar lavage fluid (BALF) in children with Mycoplasma pneumoniae pneumonia (MPP).

METHODSThe levels of TNF-α, IL-6 and IL-10 in BALF were measured using ELISA in children with MPP at acute stage (n=45) and at remission stage (n=30). Twenty children without lung lesions severed as the control group.

RESULTSThe TNF-α, IL-6 and IL-10 levels in BALF were higher in children with MPP at acute stage than those in the control group (P<0.05). The levels of TNF-α and IL-6 in BALF at remission stage were reduced to the levels similar to the control group and were significantly lower than those at the acute stage in children with MPP. However, the levels of IL-10 in BALF remained at higher levels at remission stage in children with MPP.

CONCLUSIONSThe levels of TNF-α, IL-6 and IL-10 in BALF increase in children with MPP at acute stage, suggesting that the cytokines may be involved in the pathogenesis of MPP.

Adolescent ; Bronchoalveolar Lavage Fluid ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Interleukin-10 ; analysis ; Interleukin-6 ; analysis ; Male ; Pneumonia, Mycoplasma ; etiology ; immunology ; Tumor Necrosis Factor-alpha ; analysis

BACKGROUNDGenetic association studies on populations of European origin have identified the DCDC2 gene as a susceptibility locus for developmental dyslexia. Here, we sought to investigate the association of DCDC2 polymorphisms with developmental dyslexia in children of Han Chinese origin.

METHODSWe undertook a case-control genetic association study on 76 dyslexic children and 79 non-dyslexic matched controls. We isolated DNA from oral mucosal cell samples and genotyped two DCDC2 coding-sequence single nucleotide polymorphisms, rs2274305 and rs6456593, in each sample using SNaPshot single nucleotide extension. We compared the allele and genotype frequencies between the groups using the χ(2) test and analyzed the relationship between dyslexia and the polymorphism at both loci using unconditional logistic regression. We also predicted haplotypes and compared their frequencies between the two groups.

RESULTSThe differences in the genotype distribution and the allelic genes of the two single nucleotide luci of the DCDC2 gene, rs2274305 and rs6456593, between the two dyslexic and non-dyslexic groups were statistically meaningless (P > 0.05). The differences in the haplotype distributions of the DCDC2 gene between the dyslexic and normal group were statistically meaningless (P > 0.05).

CONCLUSIONThe DCDC2 gene may not be a susceptibility factor for developmental dyslexia among the Han Chinese. However, methodological issues may have prevented the detection of positive associations.

Asian Continental Ancestry Group ; Child ; Dyslexia ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Haplotypes ; genetics ; Humans ; Male ; Microtubule-Associated Proteins ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; genetics

OBJECTIVETo investigate the feasibility and benefits of co-culture of cryopreserved islets with small intestinal submucosa (SIS).

METHODSPurified rat islets cryopreserved for one month were divided into SIS group and control group, and after culture in standard islet culture media RPMI1640 for 1 week, the morphology and function of the islets were assessed.

RESULTSThe SIS protects the fragile islets from damage by cryopreservation, and increased the recovery from (60.6-/+3.3)% to (91.7-/+1.8) % (P<0.05). Compared with the control group, incubation of the islets of the SIS group in high-glucose (16.7 mmol/L) solution resulted in significantly enhanced insulin secretion (23.7-/+1.6 vs 12.5-/+1.1 mU/L, P<0.05). When the islets were incubated in high-glucose solution containing theophylline, the calculated stimulation index of SIS group was about 3-fold higher than that of the control group.

CONCLUSIONCo-culture of cryopreserved rat islets with SIS can increase the recovery of islet cells and improve their function.

Animals ; Coculture Techniques ; Cryopreservation ; methods ; Glucose ; pharmacology ; Insulin ; secretion ; Intestinal Mucosa ; cytology ; drug effects ; physiology ; Intestine, Small ; cytology ; drug effects ; physiology ; Islets of Langerhans ; cytology ; drug effects ; physiology ; Male ; Rats ; Rats, Wistar ; Theophylline ; pharmacology

OBJECTIVE: To explore the genetic basis of a fetus with ventricular septal defect (VSD) by using modified noninvasive prenatal testing (NIPT) for the detection of microdeletion syndromes. METHODS: Chromosomal karyotypes of the fetus and its parents were analyzed by G-banding technique. Next generation sequencing (NGS) was used to detect genomic copy number variations (CNVs) in cell-free fetal DNA. The results were verified by fluorescence in situ hybridization (FISH). RESULTS: The fetus and its parents all had a normal karyotype at 320-400 band level. NGS revealed a deletion of 1.30 Mb at 7q11.23 in the fetus, with a 93% overlap with that of Williams-Beuren syndrome (WBS). The father also had a deletion of 1.42 Mb at 7q11.23, with a 99% overlap with that of WBS. Modified NIPT also detected the 1.30 Mb deletion at 7q11.23 in the fetus. The result of FISH has confirmed the above results. CONCLUSION It is necessary to carry out genetic testing on fetuses with VSD. NGS can detect fetal microdeletion syndromes and help to trace their parental origin. The modified NIPT for fetal chromosomal microdeletions/microduplication syndromes is highly accurate.
DNA Copy Number Variations ; Female ; Genetic Testing ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Pregnancy ; Prenatal Diagnosis ; Williams Syndrome

Objective To estimate the real number of novel influenza A(H1N1 ) infection in Beijing, 2009. Methods A multiplier model (Impact 2009 v 1.0 software) based on Monte Carlo approach was used to estimate the real number of novel influenza A (H1N1 ) based on the number of influenza-like illness (ILI) cases, novel influenza A(H1N1 ) positive rate among ILI cases and rate on clinical visit of ILIs in secondary and tertiary hospitals. Results There were 1.80 million (90%CI: 1.46-2.30) estimated novel influenza A (H1N1) cases in 2009 in Beijing with the rate of infection as 11.0%. One reported case would represent 167 real infections. The highest age groups of infection were 0-4 years and 5-14 years, being 32.5% and 33.3%, respectively. Conclusion Laboratory-confirmed infections with novel influenza A (H1N1 ) only represented a fraction of the total cases in a population, suggesting that it was imperative to estimate the real number of novel influenza A (H1N1) infection.