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author:("WANG, hongyu")
1.  Stability of T Cell Phenotype and Functional Assays Following Heparinized Umbilical Cord Blood Collection 
Umbilical cord blood has been used for a wide variety of immunologic investigations including assessments of developmental perturbations by antenatal exposures. Recent advances in multiparameter flow cytometry have allowed finer characterization of lymphocyte phenotype and function, revealing important differences between the fetal and adult immune systems. The degree of variability between human subjects confounds the ability to draw firm conclusions. Artifacts resulting from processing techniques exacerbate this variability. The unpredictable nature of deliveries, especially of premature infants, makes it difficult to control variables such as timing of umbilical cord mononuclear cell (UCMC) isolation and method of collection. Additionally, in multicenter studies dependent on central processing, delays are inevitable. However, little available literature describes systematic testing of the degree to which processing variations affect UCMC phenotype and function. Using multiparameter flow cytometry, we tested the effect of collection technique and length of time prior to UCMC isolation on T cell phenotype and function, with the goal of creating a standardized operating procedure for a multicenter investigation. The study also provides a benchmark data set including extensive surface and functional phenotyping of umbilical cord T cells. UCMC isolation delay of up to 24 h produced similar T cell phenotype and function as tested by in vitro SEB stimulation. There were few statistically significant differences between time points based on data medians. We conclude that, for the purpose of immunologic investigations, a 24-h time delay from sample collection to mononuclear cell isolation does not introduce a significant degree of variation in T cell phenotype and function when adhering to strict standard operating procedures.
doi:10.1002/cyto.a.22203
PMCID: PMC4029066  PMID: 23027690
umbilical cord blood; T-lymphocytes; phenotype; neonate; immunologic techniques; cell isolation; flow cytometry
2.  Association between Neonatal Iron Overload and Early Human Brain Development in Premature Infants 
Early human development  2012;88(8):583-587.
Background
Emerging evidence suggests that excess iron may be detrimental for brain development. However, little is known regarding the association between neonatal iron overload and subsequent neurodevelopment during infancy in vulnerable premature infants.
Aims
To evaluate the association between neonatal iron overload and neurodevelopment in premature infants.
Study Design
Prospective cohort study
Subjects
24–32 weeks gestational age infants who had serum ferritin (SF) measured at 34–35 weeks post-menstrual age (PMA) and did not meet exclusion criteria: SF < 76 ng/ml, toxoplasmosis, syphilis, rubella, cytomegalovirus, herpes infections, chromosomal disorders, or cranio-facial anomalies were eligible. In addition, infants with sepsis or elevated C-reactive protein within 10 days before their SF measurement were excluded.
Outcome Measures
Infants were evaluated for neurodevelopmental outcome at 8–12 months of age and were deemed to have neurodevelopmental impairment if they had one or more of the following: mental developmental index < 70, abnormal neurological examination, bilateral blindness, bilateral deafness, or required occupational, physical, or speech therapy.
Results
95 infants were studied. 70 had normal iron status (SF 76–400 ng/ml) while 25 were deemed to have iron overload (SF >400 ng/ml) at 34–35 weeks PMA. There was a marginal increase in neurodevelopmental impairment among infants with iron overload compared to infants with normal iron status (64% vs. 41%, p = 0.05). However, after controlling for confounders, iron overload was not associated with neurodevelopmental impairment (Adjusted OR 0.71, 95% CI, 0.21 – 2.5).
Conclusion
Modest neonatal iron overload is not associated with neurodevelopmental impairment during infancy in premature infants.
doi:10.1016/j.earlhumdev.2011.12.030
PMCID: PMC3677745  PMID: 22349188
premature infants; neurodevelopmental impairment; oxidative stress; iron overload; latent iron deficiency
3.  Associations among Depression, Perceived Self-Efficacy and Immune Function and Health in Preadolescent Children 
Development and psychopathology  2011;23(4):1139-1147.
Experimental animal studies and adult research consistently show that stress exposure and/or psychological symptoms are associated with poorer health and immune function. The application to children is not yet clear, however, and we lack developmental models for studies in this area. The objective of this paper was to test the hypothesis that self-reported self-efficacy and depression, two markers of psychological well-being in children, would predict immunity and rate of illnesses. The data are based on a prospective study of 141 healthy, normally developing children aged 7 – 13 years who were recruited from an ambulatory pediatric setting. Children completed self-efficacy and depression measures and had blood obtained for IL-6 plasma levels and natural killer (NK) cell functional assays on three occasions, six months apart. Parents maintained weekly child illness diaries over one year, using a thermometer to record fever. Parent psychiatric symptoms and income were used as covariates. Results indicated that, across the three occasions of measurement collected over the one-year period, higher perceived self-efficacy was significantly associated with lower plasma IL-6 concentrations. There was no overall main effect of depressive symptoms on immune measures; however, for older girls, higher depression was associated with elevated NK cell cytotoxicity and an increased rate of total illnesses and febrile illnesses. The findings provide some of the first evidence that psychological processes are associated with immunity and health in a normally developing sample of pre-adolescents. Furthermore, the pattern of results suggests a modified model of a link between psychological well-being and immunological processes in children. These results build on and expand research on the notion of allostatic load, and develop a groundwork for developmental studies in this area.
doi:10.1017/S0954579411000526
PMCID: PMC3605886  PMID: 22018086
Self efficacy; depression; interleukin 6; children’s health
4.  Randomized Controlled Trial to Improve Care for Urban Children With Asthma 
Objective
To evaluate the impact of the School-Based Asthma Therapy trial on asthma symptoms among urban children with persistent asthma.
Design
Randomized trial, with children stratified by smoke exposure in the home and randomized to a school-based care group or a usual care control group.
Setting
Rochester, New York.
Participants
Children aged 3 to 10 years with persistent asthma.
Interventions
Directly observed administration of daily preventive asthma medications by school nurses (with dose adjustments according to National Heart, Lung, and Blood Institute Expert Panel guidelines) and a home-based environmental tobacco smoke reduction program for smoke-exposed children, using motivational interviewing.
Main Outcome Measure
Mean number of symptom-free days per 2 weeks during the peak winter season (November-February), assessed by blinded interviews.
Results
We enrolled 530 children (74% participation rate). During the peak winter season, children receiving preventive medications through school had significantly more symptom-free days compared with children in the control group (adjusted difference=0.92 days per 2 weeks; 95% confidence interval, 0.50-1.33) and also had fewer nighttime symptoms, less rescue medication use, and fewer days with limited activity (allP<.01). Children in the treatment group also were less likely than those in the control group to have an exacerbation requiring treatment with prednisone (12% vs 18%, respectively; relative risk=0.64; 95% confidence interval, 0.41-1.00). Stratified analyses showed positive intervention effects even for children with smoke exposure (n=285; mean symptom-free days per 2 weeks: 11.6 for children in the treatment group vs 10.9 for those in the control group; difference=0.96 days per 2 weeks; 95% confidence interval, 0.39-1.52).
Conclusions
The School-Based Asthma Therapy intervention significantly improved symptoms among urban children with persistent asthma. This program could serve as a model for improved asthma care in urban communities.
doi:10.1001/archpediatrics.2011.1
PMCID: PMC3600609  PMID: 21383275
5.  Immunogenicity of Trivalent Influenza Vaccine in Extremely-Low-Birth-Weight, Premature versus Term Infants 
Background
Influenza vaccine immunogenicity in premature infants is incompletely characterized.
Objective
To assess the immunogenicity of trivalent, inactivated influenza vaccine (TIV) in extremely low-birth-weight (ELBW, ≤1000 grams birth weight), premature (<30 weeks gestation) infants. We hypothesized that geometric mean titers (GMT) of influenza antibody would be lower in premature than in full-term (≥37 week) infants.
Design/Methods
In this prospective, multicenter study, former premature and full-term infants ages, 6–17 months, received 2 doses of TIV during the 2006–7 or 2007–8 influenza seasons. Sera were drawn before dose 1 and 4–6 weeks after dose 2. Antibody was measured by hemagglutination inhibition.
Results
Over two years, 41 premature and 42 full-term infants were enrolled; 36 and 33 of these infants, respectively, had post-vaccination titers available. Premature infants weighed less (mean 1.3 – 1.8 kg difference) at the time of immunization than full-term infants. Pre-vaccination titers did not differ between groups. Premature infants had higher post-vaccination antibody GMT than full-term infants to H1 (2006–7, 1:513 v. 1:91, P=0.03; 2007–8, 1:363 v. 1:189, P=0.02) and B/Victoria (2006–7, 1:51 v. 1:10, P=0.02). More premature than full-term infants had antibody titers ≥ 1:32 to B/Victoria (85% v. 60%, p=0.04) in 2007–8. Two (5%) premature and 8 (19%) full-term infants had adverse events, primarily fever, within 72 hours after vaccination. No child had medically-diagnosed influenza.
Conclusions
Former premature infants had antibody responses to two TIV doses greater than or equal to those of full-term children. Two TIV doses are immunogenic and well tolerated in ELBW, premature infants 6–17 months old.
doi:10.1097/INF.0b013e31820c1fdf
PMCID: PMC3090695  PMID: 21273938
Premature infant; very low birth weight infant; influenza vaccines; immunization; vaccines
6.  Restrictive Anorexia Nervosa And Set-Shifting in Adolescents: A Biobehavioral Interface 
The Journal of Adolescent Health  2011;49(1):99-101.
PURPOSE
Set-shifting is a neurocognitive concept defined as the ability to switch tasks flexibly. Set-shifting scores are worse in adults with Restrictive Anorexia Nervosa (AN-R) than in controls. Adolescence is a developmental period when young people must respond flexibly to new situations. The purpose of this study is to compare the set-shifting scores of 24 adolescent females with AN-R and 37 matched normal adolescent controls (ages 14–20).
METHODS
Methods included sociodemographic, psychological and biologic data, and neurocognitive testing using the Behavior Rating of Executive Function – Self- and Parent-Reports, the Cambridge Neuropsychological Automated Battery, and the Wisconsin Card-Sorting Test. Statistical analyses included t-tests, multiple analysis of variance, and correlations.
RESULTS
Study and control subjects were similar on sociodemographic data and intelligence quotient. There were differences in Body Mass Index and the Eating Disorder-3 evaluation. Significant differences in the composite score of set-shifting between the study and control groups were found using multiple analysis of variance.
CONCLUSIONS
Adolescent females with AN-R had significantly worse set-shifting scores than did control subjects. Future studies of adolescent AN-R subjects should include biologic (fMRI) and neurocognitive measures to determine the mechanisms at the brain-behavioral interface so that treatment can be directed specifically to set-shifting deficits.
doi:10.1016/j.jadohealth.2010.11.259
PMCID: PMC3286875  PMID: 21700167
7.  Parental Assessment of Executive Function and Internalizing and Externalizing Behavior in Primary Hypertension after Antihypertensive Therapy 
The Journal of pediatrics  2010;157(1):114-119.
Objective
To determine the change in parental ratings of executive function and behavior in children with primary hypertension following antihypertensive therapy.
Study design
Parents of untreated hypertensive subjects and controls completed the Behavior Rating Inventory of Executive Function (BRIEF) to assess behavioral correlates of executive function and the Child Behavior Checklist (CBCL) to assess internalizing and externalizing behaviors. Hypertensive subjects subsequently received antihypertensive therapy to achieve casual BP < 95th percentile. After 12 months, all parents again completed the BRIEF and CBCL.
Results
Twenty-two subjects with hypertension and 25 normotensive control subjects had both baseline and 12-month assessments. Hypertensive subject’s blood pressure improved (24-hr systolic BP load: mean baseline vs. 12-months, 60 vs. 25%, p < 0.001). Parent ratings of executive function improved from baseline to 12-months in the hypertensives (BRIEF Global Executive Composite T-score, Δ = −5.9, p = 0.001) but not in the normotensive controls (Δ = −0.36, p = 0.83). In contrast, T-scores on the Child Behavior Checklist Internalizing and Externalizing summary scales did not change significantly from baseline to 12-months in either hypertensive or control subjects.
Conclusions
Children with hypertension demonstrated improvement in parental ratings of executive function after 12 months of antihypertensive therapy.
doi:10.1016/j.jpeds.2009.12.053
PMCID: PMC2904985  PMID: 20227722
Neurocognitive function; blood pressure; treatment
8.  Diagnostic Assays for Active Infection with Human Herpesvirus 6 (HHV-6) 
Background
Human herpesvirus 6 (HHV-6) causes ubiquitous infection in early childhood with lifelong latency or persistence. Reactivation of HHV-6 has been associated with multiple diseases including encephalitis. Chromosomal integration of HHV-6 also occurs. Previous studies have suggested that the detection of HHV-6 DNA in plasma is an accurate marker of active viral replication.
Objective
We sought to determine whether PCR assays on plasma could correctly differentiate between primary HHV-6 infection, chromosomal integration of HHV-6 and latent HHV-6 infection.
Study Design
We performed qualitative PCR, real-time quantitative PCR (RQ-PCR), and reverse-transcriptase PCR (RT-PCR) assays on samples of peripheral and cord blood mononuclear cells, as well as plasma, from groups of subjects with well defined HHV-6 infection, including subjects with chromosomally integrated HHV-6.
Results and Conclusions
The detection of HHV-6 DNA in plasma was 92% sensitive compared to viral isolation for the identification of primary infection with HHV-6. All plasma samples from infants with chromosomally integrated HHV-6 had HHV-6 DNA detectable in plasma while only 5.6% were positive by RT-PCR. The specificity of plasma PCR for active replication of HHV-6 was 84% compared to viral culture while the specificity of RT-PCR was 98%. Our results demonstrate that qualitative or quantitative PCR of plasma is insufficient to distinguish between active viral replication and chromosomal integration with HHV-6. We found a higher specificity of RT-PCR performed on PBMC samples compared to PCR or RQ-PCR performed on plasma when evaluating samples for active HHV-6 replication.
doi:10.1016/j.jcv.2010.02.007
PMCID: PMC2855742  PMID: 20211581
Human herpesvirus 6; chromosomally integrated HHV-6 (CI-HHV-6); polymerase chain reaction; real time quantitative polymerase chain reaction (RQ-PCR); reverse transcriptase polymerase chain reaction (RT-PCR)
9.  Perceptions of Body Image by Persons With Prader-Willi Syndrome and Their Parents 
Prader-Willi syndrome is a genetic disorder characterized by obesity. The Figure Rating Scale (Stunkard, Sorensen, & Schulsinger, 1983) was completed by 43 individuals with this syndrome to determine their level of dissatisfaction with their body. Their parents also completed this scale regarding their child to determine whether they were dissatisfied with their child’s body status. Results showed that individuals with Prader-Willi syndrome were dissatisfied with their body. Parents also were dissatisfied with their child’s body. Results of this study demonstrate that the responses of persons with Prader-Willi syndrome on the Figure Rating Scale show significant discrepancies between how they think they look and how they wished they looked.
doi:10.1352/1944-7558-115.1.43
PMCID: PMC2865429  PMID: 20025358
10.  Parental Assessments of Internalizing and Externalizing Behavior and Executive Function in Children with Primary Hypertension 
The Journal of pediatrics  2008;154(2):207-212.
Objective
To determine the relations between hypertension and parental ratings of behavior and executive functions in children with primary hypertension and to examine the potential moderating influence of obesity.
Study Design
Hypertensive and normotensive control groups were matched for age, sex, race, intelligence quotient, maternal education, household income, and obesity. Parents completed the Child Behavior Checklist (CBCL) to assess Internalizing and Externalizing problems and the Behavior Rating Inventory of Executive Function (BRIEF) to assess behavioral correlates of executive function.
Results
Thirty-two hypertensive subjects and 32 normotensive controls (10 –18y) were enrolled. On the CBCL, hypertensives had higher Internalizing T-scores (53 vs. 44.5, p = 0.02) with 37% falling within the clinically significant range vs. 6% of controls (p = 0.005). Internalizing score increased with increasing BMI percentile in hypertensive, but not normotensive, subjects. Hypertensives had worse BRIEF Global Executive Composite (GEC) T-scores compared with controls (50 vs. 43, p = 0.009).
Conclusions
Children with both hypertension and obesity demonstrate higher rates of clinically significant internalizing problems, and hypertensives (irrespective of obesity) demonstrate lower parental ratings of executive function compared with normotensive controls.
doi:10.1016/j.jpeds.2008.08.017
PMCID: PMC2633107  PMID: 18823913
Neurocognitive function; anxiety; depression; blood pressure
11.  The Associations between Psychosocial Stress and the Frequency of Illness, and Innate and Adaptive Immune Function in Children 
Brain, behavior, and immunity  2008;22(6):933-940.
Objective
Family processes have a substantial impact on children’s social and emotional well-being, but little is known about the effects of family stress on children’s physical health. To begin to identify potential links between family stress and health in children, we examined associations between specific aspects of family psychosocial stress and the frequency of illnesses in children, measures of innate and adaptive immune function, and human herpesvirus 6 (HHV-6) reactivation.
Study Design
Prospective study of 169 ambulatory school-age children and parents. Parents completed multiple assessments of stress at 7 sequential six-month visits and maintained weekly illness diaries for their children over three years using a thermometer to record fever. Children had blood obtained for HHV-6 and immune function studies at each visit including natural killer (NK) cell function and the percentage of CD4 and CD8 cells associated with immune control of cytomegalovirus (CMV).
Results
Parental psychiatric symptoms were associated with a higher frequency of illnesses: for each 1 unit increase in symptom score children had an increased 1-year rate of total illnesses of 40% (rate ratio, 1.40; 95% CI, 1.06–1.85) and febrile illnesses of 77% (rate ratio, 1.77, 95% CI, 1.00–3.13). Parental psychiatric symptom scores were also associated with enhanced NK cell function (estimate, 0.15; 95% CI, 0.05–0.26) and increased percentages of CD8+CD28-CD57+ cells in the blood of CMV seropositive children (estimate, 2.57; 95% CI, 0.36–4.79). HHV-6 reactivation was not detected.
Conclusions
There is an association between specific psychosocial stress exposure and rates of illness and immune function in normally developing children.
doi:10.1016/j.bbi.2008.01.007
PMCID: PMC2516370  PMID: 18308510
12.  Left ventricular mass index in children with white coat hypertension 
The Journal of pediatrics  2008;153(1):50-54.
Objective
To determine if children with white coat hypertension (WCH) have evidence of target-organ damage by comparing left ventricular mass index (LVMI) of subjects with WCH to that of matched normotensive and hypertensive controls.
Study design
Each WCH subject was matched by body mass index (± 10%), age (± 1 year), and sex to a normotensive control and to a hypertensive control. Echocardiograms were reviewed to determine LVMI for each subject. These triple matches were analyzed using repeated measures analysis of variance to detect differences in LVMI between the three groups.
Results
Twenty-seven matched triplets were established. The groups were comparable for sex, age, and body mass index (BMI). Mean LVMI was 29.2, 32.3, and 35.1 g/m2.7, for normotensives, WCH, and sustained hypertensives, respectively (normotensive vs. WCH, p = 0.028; WCH vs. sustained hypertensive, p = 0.07). Left ventricular hypertrophy was not present in any subject in the normotensive or WCH groups, but was present in 26% of the sustained hypertensive subjects (p < 0.001).
Conclusions
After controlling closely for BMI, children with WCH had a LVMI which was intermediate between that of normotensives and sustained hypertensives, suggesting that WCH may be associated with hypertensive end-organ effects.
doi:10.1016/j.jpeds.2008.01.025
PMCID: PMC2516747  PMID: 18571535

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