A comparative analysis of neonatal critical illness score and score for neonatal acute physiology, perinatal extension, version II / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the accuracy and clinical utility of neonatal critical illness score (NCIS) and score for neonatal acute physiology, perinatal extension, version II (SNAPPE-II) in predicting the "dead and abandoned" risk in critically ill neonates.</p><p><b>METHODS</b>A total of 269 critically ill neonates were divided into two groups according to their prognosis: dead/abandoned and improved/cured. The accuracy of these two scoring systems, NCIS and SNAPPE-II, in predicting the "dead and abandoned" risk was compared.</p><p><b>RESULTS</b>The dead/abandoned group had a significantly higher SNAPPE-II score than the improved/cured group (P<0.001), while there was no significant difference in the NCIS score between the two groups (P=0.091). The children who were in line with the individual indicator in the NCIS results had a significantly higher "dead and abandoned" risk than those who were not (P=0.005).</p><p><b>CONCLUSIONS</b>SNAPPE-II is more accurate in early prediction of the "dead and abandoned" risk in critically ill neonates compared with NCIS. NCIS has the ability to predict the "dead and abandoned" risk in children in line with the individual indicator.</p>

Prospective clinical study of radix astragali and its compound prescription for treatment of β-thalassemia in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of radix astragali and its compound prescription for treatment of β-thalassemia in children.</p><p><b>METHODS</b>This study was a randomized, controlled, double-blind clinical trial. Fifty-seven children with β-thalassemia were randomly assigned to radix astragali, compound prescription (radix astragali+ codonopsis pilosula + tortoise plastron) and placebo control groups after stratifying the patients according to disease type (intermedia and major). The parameters of hematology and safety were assessed after 12 weeks of treatment.</p><p><b>RESULTS</b>After 12 weeks of treatment, the mean Hb elevation levels in children with β-thalassemia intermedia from the compound prescription and the radix astragali groups were 1.21±1.12 and 1.05±0.80 g/dL respectively compared with -(0.28±0.51) g/dL in the placebo control group (P<0.01). Mean Hb levels in the compound prescription and radix astragali groups were significantly higher than in the placebo control group (P<0.05). Therapy with both radix astragali and its compound prescription increased fetal hemoglobin, red blood cell, mean corpuscular hemoglobin and reticulocyte levels in children with β-thalassemia intermedia. The total effective rates were 64% and 62% in children with β-thalassemia intermedia from the compound prescription and radix astragali groups respectively, which was significantly higher than in the placebo control group (9%; P<0.01). Therapy with radix astragali or its compound prescription in children with β-thalassemia major had similar but less favourable effects than the same therapy in children with β-thalassemia intermedia. White blood cell, neutrophil, platelet and hepatic and renal functions were not adversely affected by the medicines.</p><p><b>CONCLUSIONS</b>Therapy with radix astragali or its compound prescription is effective and safe in children with β-thalassemia.</p>

Preparation of polyclonal antibody of recombinant human thioredoxin-1 and its protective effects on neonatal rats with endotoxemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To clone the gene human thioredoxin 1 (hTrx-1) expressing its protein in the E.coli expression system and to obtain its polyclonal antibody, and to study the protective effects of hTrx-1 on neonatal rats with endotoxemia.</p><p><b>METHODS</b>DNA encoding hTrx-1 from fetal liver cells was isolated by RT-PCR. The hTrx-1 was cloned to the prokaryotic expression plasmid PET-28a to induce its protein expression in the E.coli expression system. The purified hTrx-1 was injected into rats to prepare polyclonal antibody. Newborn Sprague-Dawley rats were randomly assigned to three groups: control, lipopolysaccharide (LPS) and hTrx-1 (n=12 each). The control and the LPS groups were intraperitoneally injected with normal saline and LPS (5 mg/kg), respectively. The hTrx-1 group received an intraperitoneal injection of hTrx-1 (10 mg/kg) 30 minutes before LPS injection. The mortality rate 24 hrs after injection was compared between the three groups.</p><p><b>RESULTS</b>The prokaryotic expression plasmid PET-28a-hTrx-1 was constructed. The hTrx-1 protein was expressed and purified. The polyclonal antibody of hTrx-1 with the titer of 1∶51200 was prepared. The mortality rate of the control, LPS and hTrx-1 groups was 0, 67% and 17%, respectively (χ2=14.400, P<0.01).</p><p><b>CONCLUSIONS</b>The polyclonal antibody of hTrx-1 is prepared successfully. The hTrx-1 protein has protective effects on neonatal rats with endotoxiamia.</p>

Effects of sodium butyrate on acetylation of histone in gamma-globin gene promoter regions in K562 cells: a study using chromatin immunoprecipitation / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop a real-time PCR-based chromatin immunoprecipitation (ChIP) assay for determining the effect of sodium butyrate on acetylation of histone in gamma-globin gene promoter regions in K562 cells.</p><p><b>METHODS</b>K562 cells were grown in the presence or absence of 0.5 mmol/L sodium butyrate for 48 h, and 1=10(7) cells per group were used for real-time PCR-based ChIP with anti-acetylated histone H3 or H4 antibodies. The levels of acetylated histone H3 and H4 (acH3 and acH4) in Ggamma- and Agamma-globin gene promoter regions were measured.</p><p><b>RESULTS</b>In the K562 cells with sodium butyrate treatment or without any treatment, the levels of acH3 or acH4 in Ggamma- or Agamma-globin gene promoter were higher than that in the necdin gene (negative control). Compared with the untreated K562 cells, the cells treated with 0.5 mmol/L sodium butyrate showed a 3.1-fold or 2.6-fold increase in acH3 or acH4 in Ggamma-globin gene promoter region, with also a 3.7-fold or 3.2-fold increase in acH3 or acH4 in Agamma-globin gene promoter region, respectively (P<0.01).</p><p><b>CONCLUSION</b>We have successfully developed a real-time PCR-based ChIP assay for analyzing the acetylation of histone H3 and H4 in gamma-globin gene promoter regions. Our results support the role of sodium butyrate in increasing the level of acetylated histone in gamma-globin gene promoter regions.</p>

Risk factors for adverse neonatal outcomes in 254 twins / 中国当代儿科杂志

<p><b>OBJECTIVE</b>The aim of this study was to identify the risk factors for adverse neonatal outcome in twins in order to provide a basis for the improvement of the survival and neonatal outcomes of twins.</p><p><b>METHODS</b>Data from 254 twins admitted to Nanfang Hospital of Southern Medical University From January 2005 to December 2009 were retrospectively studied. Risk factors for adverse neonatal outcomes were assessed by logistic regression analysis.</p><p><b>RESULTS</b>Of the 254 twins, 84 (33.1%) had an adverse outcome, including 10 (3.9%) neonatal deaths. Logistic regression analysis demonstrated that gestational age (≤34 weeks), cord abnormalities, meconium-stained amniotic fluid and 5-min Apgar scores (≤7) were independent risk factors for adverse neonatal outcomes (OR=4.434, 4.731, 3.424, 18.958, respectively; P=0.021, 0.001, 0.037, 0.011, respectively). Conception by assisted reproductive technology was shown as a protective factor for adverse neonatal outcomes (OR=0.389, P=0.037).</p><p><b>CONCLUSIONS</b>The twins with gestational age ≤34 weeks, cord abnormalities, meconium-stained amniotic fluid or 5-min Apgar scores (≤7) are subject to adverse neonatal outcome.</p>

Effect of low-dose hydroxyurea with sodium butyrate on globin gene expression in human erythroid progenitor cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of combined use of low-dose hydroxyurea (HU) and sodium butyrate (NaB) on the expression of 7 globin genes (zeta, alpha, epsilon, Ggamma, Agamma, delta, and beta) in human erythroid progenitor cells.</p><p><b>METHODS</b>Human erythroid progenitor cells were cultured using a two-step liquid culture system and treated with HU and NaB either alone or in combination. The inhibitory effects of the agents on the cell growth were monitored with trypan blue exclusion assay, and the changes in the mRNA of the 7 globin genes were detected using RT-PCR.</p><p><b>RESULTS</b>Low-dose HU combined with NaB resulted in significantly lower inhibition rate of the erythroid progenitor cells than routine dose HU and NaB used alone (28.56% and 38.80%, respectively, P<0.05). Compared with untreated cells (0.653-/+0.092 and 0.515-/+0.048), HU combined with NaB significantly increased the expression of Ggamma-and Agamma- mRNA (1.203-/+0.018 and 0.915-/+0.088, respectively, P<0.05), and HU and NaB used alone produced similar effects (1.305-/+0.016 and 0.956-/+0.029 for HU, and 1.193-/+0.070 and 0.883-/+0.012 for NaB, P>0.05). HU and NaB, either used alone or in combination or at different doses, caused no significant changes in the other globin genes (zeta, alpha, epsilon, delta and beta) (P>0.05).</p><p><b>CONCLUSION</b>Low-dose HU combined with NaB can up-regulate gamma globin gene expression, especially Ggamma-mRNA expression, to decrease the growth inhibition on human erythroid progenitor cells in vitro, but produces no significant effect on the expressions of zeta, alpha, epsilon, delta and beta genes.</p>

Astragalus polysaccharides-induced gamma-globin mRNA expression in K562 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of Astragalus polysaccharides (APS) in inducing the mRNA expression of Agamma- and Ggamma-globin in K562 cells.</p><p><b>METHODS</b>K562 cells were treated with APS at the concentration of 150, 300, and 450 mg/L, with Na-butyrate (NaB)-treated cells serving as the positive control and untreated cells as the blank control. Benzidine staining was used to examine the changes in hemoglobin synthesis in K562 cells after the treatments, and RT-PCR was employed to investigate the mRAN expression of Agamma- and Ggamma-globin.</p><p><b>RESULTS</b>Compared with the untreated cells, APS treatment (300 mg/L) for 48 h resulted in a significant increase of the percentages of benzidine-positive cells from (4.37-/+0.58)% to (15.67-/+1.80)%, and also in significantly increased expression of Agamma-globin and Ggamma-globin mRNAs by 3.59-/+0.16 and 5.02-/+0.81 folds, respectively (P=0.000).</p><p><b>CONCLUSION</b>APS potently enhances the mRNA expression of Agamma- and Ggamma-globin in K562 cells and warrants further evaluation as a potential therapeutic agent for beta-thalassemia.</p>

Adenovirus infection in hospitalized children with pneumonia in Guangzhou from 2005 to 2007 / 南方医科大学学报

<p><b>OBJECTIVE</b>To understand the characteristics of adenovirus infection in hospitalized children with pneumonia in Guangzhou area.</p><p><b>METHODS</b>The infection rate, hospitalization time and hospitalization expenses of adenovirus-infected hospitalized children with pneumonia in Guangzhou area from 2005 to 2007 were analyzed.</p><p><b>RESULTS</b>The total adenovirus infection rate was 6.04% in these children, with a male to female ratio of 1.47:1, showing significantly higher infection rate in female (7.92%) than in male patients (5.21%, P<0.05). The hospital stay and hospitalization costs between male and female children showed no significant difference (P>0.05). Adenovirus-infected children from birth to six years old accounted for 90.50% of the total adenovirus-infected children, and the infection rate in 0 to 1-year-old children (3.71%) was significantly lower than that in elder children (P<0.05). Although the infection rate in winter (8.44%) was significantly higher than that in the other seasons (P<0.05), the cases from March to August accounted for 60.11% of the total infected cases. Furthermore, the infection rate in 2007 (4.31%) was significantly lower than that in 2005 and 2006 (7.11% and 6.71%, respectively, P<0.05).</p><p><b>CONCLUSION</b>Adenovirus infection is an important pathogen in hospitalized children with pneumonia in Guangzhou area, and the infection rates differed between gender, age, season and the years.</p>

Gamma-globin synthesis in K562 cells induced with Tortois plastron, Astragali, Salviae miltiorrhizae and Codonopsis pilosulae / 中国实验血液学杂志

This study was purposed to investigate the effects of tortois plastron, astragali, salviae miltiorrhizae and codonopsis pilosulae on gamma-globin gene synthesis in K562 cells in vitro. Benzidine staining was used to clarify the dose-and time-dependent effects of tortois plastron, astragali, salviae miltiorrhizae and codonopsis pilosulae on hemoglobin synthesis in K562 cells and Western blotting was performed to determine the level of hemoglobin F (alpha(2)gamma(2)). The results indicated that the K562 cells treated with 4 kinds of traditional Chinese medicine had different stain rates of benzidine for: 23.5% (tortois plastron), 19.8% (astragali), 15.8% (salviae miltiorrhizae) and 14.5% (codonopsis pilosulae) at 6 days after the treatment. Western blot indicated that synthesis of HbF increased. It is concluded that tortois plastron, astragali, salviae miltiorrhizae and codonopsis pilosulae enhance globins-gamma synthesis level and increase hemoglobin F level in K562 cells, the effect of which resembles that of sodium butyrate.

Rearrangement of immunoglobulin heavy chain variable region genes in human neonates / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the rearrangement of immunoglobulin (Ig) heavy chain variable region (V(H)) genes in human neonates with different gestational ages (GA).</p><p><b>METHODS</b>Peripheral blood from the neonates with GA of 27 weeks (4 cases), 28-32 weeks (9 cases), 33-36 weeks (12 cases), and 37-42 weeks (13 cases) was collected. RT-PCR was used to amplify the Ig V(H) gene, and the PCR products were separated by electrophoresis and analyzed using 6% denaturing PAGE gel.</p><p><b>RESULTS</b>All Ig V(H) family genes had several rearranged genes in each GA group, and the neonates with different GA showed no significant difference in the median molecular weight for each rearranged Ig V(H) family gene.</p><p><b>CONCLUSION</b>The neonates with GA of 27-42 weeks exhibit diversity in Ig V(H) gene rearrangement, and for the same Ig V(H) family, the median length of the arranged Ig V(H) genes is independent of the gestational age.</p>

Specific inhibition of gene expression of lung resistance-related protein by short interfering RNA / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate inhibitory effect of short interfering RNA (siRNA) on the expression of lung resistance-related protein (LRP) in leukemia cells.</p><p><b>METHODS</b>The eukaryotic vectors of LRP, pcDNA3.0/LRP, were constructed. The transfection protocol of K562 cells grown in standard conditions consisted of different combinations of pcDNA3.0/LRP, pEGFP-C1 expressing mammalian enhanced green fluorescent protein (GFP), and their gene-specific siRNAs. RT-PCR and flow cytometry were employed to evaluate the mRNA and protein expression of LRP and fluoroscopy was performed for assay of GFP expression in the transfected cells.</p><p><b>RESULTS</b>Compared with untreated K562 cells, pcDNA3.0/LRP-transfected cells showed increased LRP mRNA and protein expression and the positive cell percentage reached 30%. In the cells co-transfected with LRP gene-specific siRNA and pcDNA3.0/LRP, both LRP mRNA and protein expression decreased significantly to a level defined as negative results; the GFP expression showed no significant difference between the cells transfected with pEGFP-C1 and those co-transfected with LRP gene-specific siRNA and pEGFP-C1. LRP mRNA and protein expressions were also similar between the cells transfected with pcDNA3.0/LRP and those co-transfected with GFP gene-specific siRNA and pcDNA3.0/LRP.</p><p><b>CONCLUSIONS</b>The LRP gene-specific siRNA we designed is capable of degrading LRP mRNA and inhibiting the protein expression effectively and specifically, which shed light on the potential application of siRNA for gene-specific therapy to reverse LRP-induced multidrug resistance of leukemia cells.</p>

Analysis of therapeutic effectiveness of Nanfang ALL 99 protocol in childhood acute lymphoblastic leukemia patients / 中华儿科杂志

<p><b>OBJECTIVE</b>With more precise diagnostic criteria and risk classifications, more effective therapy administered in clinical trials, and better supportive care, the outcome of children with acute lymphoblastic leukemia (ALL) has been improved dramatically. Today, approximately 80% of children treated for this disease in developed countries enjoy long-term event free survival (EFS) and in most instances, would be cured. In this study, treatment outcome of 82 childhood ALL patients in the hospital were analyzed, and ways for how to improve the EFS rate in childhood ALL were explored.</p><p><b>METHODS</b>Eighty-two patients with ALL were enrolled into the Nanfang ALL 99 protocol which derived from German BFM ALL 95 and Hong Kong-Singapore acute lymphoblastic leukemia 97 (HK-SG ALL 97). Dexamethasone instead of hydrocortisone was used for triple intrathecal therapy. Standard at risk patients who had been irregularly treated in other hospitals for short periods of time were classified as at intermediate risk. When ANC was > or = 1.0 x 10(9)/L and platelet > or = 100 x 10(9)/L, chemotherapy was started. Life table method was used to estimate survival rate and statistical analysis was done by using software SPSS for Windows.</p><p><b>RESULTS</b>From March 1999 to September 2003, 82 childhood ALL patients were treated with the Nanfang ALL 99 protocol and 78 (95.1%) patients attained complete remission (CR) in a median time of 33 days. Out of 82 patients, 13 patients dropped out of the the Nanfang ALL 99 protocol because of financial difficulty or other reasons. Sixty nine patients were consecutively treated with the Nanfang ALL 99 protocol. The overall EFS rate at 2 years, 3 years and 5 years were 91.3%, 85.9% and 75.2%, respectively, with a median observation duration of 34 months. Three patients died of complications (4.3%). The disease relapsed in 6 patients and they died finally.</p><p><b>CONCLUSION</b>The outcome of patients treated with the Nanfang ALL 99 protocol was favorable, and the mortality rate of this chemotherapeutic protocol was low. This protocol was well tolerated by Chinese patients and therefore the protocol is worthy of application in China.</p>

Effect of Sodium Butyrate on Phosphorylation of Histone at ?-Globin Gene Promoter Regions in K562 Cells / 实用儿科临床杂志

Objective To explore the effect of sodium butyrate(NaB) on phosphorylation/ acetylation of histone H3(ph/acH3) at G?-globin gene and A?-globin gene promoter regions in K562 cells.Methods K562 cells were devided into 2 groups:K562 cells were grown in the presence or absence of 0.5 mmol?L-1NaB for 48 h [K562(NaB) group] and untreated K562 cells group(K562 group).Semi-quantitative RT-PCR was employed to measure the levels of G?-globin mRNA and A?-globin mRNA.The real time PCR-based chromatin immunoprecipitation(ChIP) was used to detect the levels of ph/acH3 at G?-globin gene and A?-globin gene promoter regions.Results Compared with the K562 group,there was a 1.4-fold(t=-149.022,P=0.000) and 1.2-fold(t=-13.363,P=0.000) increase in G?-globin mRNA and A?-globin mRNA,respectively,in K562(NaB) group.The level of ph/acH3 at G?-globin gene and A?-globin gene promoter region increased by 2.9-fold(t=-12.833,P=0.006) and 3.2-fold(t=-10.484,P=0.000),respectively,in K562(NaB) group,compared with the K562 group.The %Input value of G?-globin and A?-globin promoter fragment was 10.0-fold(P=0.000) and 9.5-fold(P=0.000) higher than that value of Necdin gene promoter fragment in the K562(NaB) group,while the %Input value of G?-globin and A?-globin promoter fragment was 3.2-fold(P=0.000) and 2.7-fold(P=0.000) higher than that value of necdin gene promoter fragment in K562 group.Conclusions NaB improves the phosphorylation and acetylation of H3 at ?-globin gene promoter regions,and this may be one of the mechanisms of expression of ?-globin genes induced by NaB.

Influence of Astragalus Polysaccharides on Fetal Hemoglobin Synthesis and Cell Proliferation in K562 Cells / 实用儿科临床杂志

Objective To explore the effects of astragalus polysaccharides(APS) on fetal hemoglobin(HbF) synthesis and cell proli-feration in K562 cells.Methods K562 cells were chosen as the cell model and cells treated with Na-butyrate(NaB) were taken as the po-sitive control.Western blot was applied to study the level of HbF expression in K562 cells and Trypan blue dye exclusion test was employed to analyze the influence of APS(150 mg/L,300 mg/L,450 mg/L)on K562 cells proliferation.Results 1.Dosage effect:when compared with untreated K562 cells,the HbF expression level increased to(1.56?0.03),(1.78?0.04) and(1.51?0.32) fold,respectively after 48 h treated with different concentrations of APS(150 mg/L,300 mg/L,450 mg/L,F=310.476 P=0).The best inducing concentration was 300 mg/L(P=0.005).2.Time course: HbF levels raised up gradually and the maximum was(2.88?0.27) fold over baseline(P=0) at 48-60 h in the presence of 300 mg/L APS.Then it went to decline.There was statistical significance of HbF expression between K562 cells treated with 300 mg/L APS or NaB [(2.88?0.27) folds,P=0].3.Effects of APS on K562 cells proliferation:the highest reduction of the cell proliferative was obtained in K562 cells cultured in the presence of 0.5 mmol/L NaB.As detected by Trypan blue exclusion met-hod,growth rate of cells stimulated by APS was affect in a dose dependent manner,and significantly higher than NaB.For example,the inhibition rate at 48 hours was 20.45% for 300 mg/L APS but 79.55% for 0.5 mmol/L NaB(P=0).Conclusion APS has ability to induce HbF synthesis in K562 cells and revealed less cells reduction than that of NaB.

Analysis of Respiratory Syncytial Virus Infection in Hospitalized Children with Pneumonia in Guangzhou Area from 2005 to 2007 / 实用儿科临床杂志

6-11 years old were 9.67%, 6.81%, 3.49% and 0.80%, respectively.Furthermore, the infection rates between each two age stages were significantly different(Pa0.05).4.Infection rates in 2005,2006 and 2007 were 4.0%, 8.92%, 8.85%,respectively.Infection rates between 2005 and 2006,2007 were significantly different(Pa

Effect of Sodium Butyrate on Erythroid Lineage by Expressional Profiles Microarray in K562 Cells / 实用儿科临床杂志

1)which represented 340 genes and 171 down-regulated(SLR

Relationship between Phosphorylation of p38 and Erythroid Differentiation of Human K562 Erythroleukemia Cells / 实用儿科临床杂志

Objective To investigate the role of directly constitutive activation of p38 mitogen-activated protein kinases(p38MAPKs)signaling in ?-globin gene expression and fetal hemoglobin(HbF)induction,and provide direct data for the relationship between phosphorylation of p38 and erythroid differentiation of human K562 erythroleukemia cells.Methods The human K562 erythroleukemia cells were transfected with pCDNA 3.1-MKK3(Glu)and pCDNA 3.1-MKK3(Ala)recombinant plasmids by lipofectamineTM 2000.Then,the stable cell lines overexpressing constitutively active p38 and constitutively inhibitive p38 activation were established by the addition of G418 to select single cell G418-resistant clones and identification with reverse transcriptase-polymerase chain reaction(RT-RCR)and Western blot assays,named K562-MKK3(Glu)and K562-MKK3(Ala)cells,respectively.Furthermore,the direct effects of constitutively active p38 on the ?-globin gene expression and HbF induction were analyzed by RT-PCR and benzidine staining,respectively.Results The results of RT-PCR and Western blot showed that there were no evident changes in the mRNA and protein levels of p38 for various cell models,but compared with K562,K562-vect,and K562-MKK3(Ala)cells,the phosphorylation of p38 and expression of ?-globin levels in K562-MKK3(Glu)cells were significantly up-regulated.The results of benzidine staining displayed that the mean percentages of positive cells stained by benzidine in K562,K562-vect,K562-MKK3(Ala),K562-MKK3(Glu)cells,and K562-MKK3(Glu)cells treated with SB203580 were(3.2?1.4)%,(3.7?1.2)%,(2.8?0.9)%,(32.6?5.3)%,and(7.8? 2.3)%(q = 7.56 P