Expression and significance of toll-like receptors 7 and 8 in brain and lung tissues of death cases caused by EV71 infection / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the significance of toll-like receptors (TLR) -7 and -8 in the pathogenesis of infection caused by Enterovirus type 71 (EV71) through measuring the expression of TLR7 and TLR8 in brain and lung tissues from the death cases caused by EV71 infection.</p><p><b>METHODS</b>Nine children who died of EV71 infection (EV71 group) were selected as study subjects, and 7 children who died of accidents or non-infectious diseases were used as the control group. Brain and lung tissues from the death cases in both groups at autopsy were collected, and immunohistochemistry was applied to detect the expression of TLR7 and TLR8 in lung and brain tissues in both groups. Integrated optical density (IOD) was applied for semi-quantitative analysis of the expression of TLR7 and TLR8.</p><p><b>RESULTS</b>Immunohistochemical results showed that the expression of TLR7 and TLR8 in lung and brain tissues was strongly positive in the EV71 group, and the IOD values in the EV71 group were also significantly higher than those in the control group (P<0.05). There was no significant difference in the expression of TLR7 and TLR8 between lung and brain tissues in the EV71 group (P>0.05).</p><p><b>CONCLUSIONS</b>TLR7 and TLR8 are highly expressed in lung and brain tissues from the patients who die of severe EV71 infection, suggesting that TLR7 and TLR8 may be involved in the pathogenesis of brain and lung damages caused by severe EV71 infection.</p>

Expression of Cysteinyl leukotriene receptor 1 in bronchial epithelial cell regulated by tumor necrosis factor-α / 中华实用儿科临床杂志

Objective To explore whether TNF-α involves in the modulation of Cysteinyl leukotriene receptor 1 (CysLT1) expression in bronchial epithelial cells.Methods The bronchial epithelial cell lines 16HBE cells were stimulated with different concentration (0.00,0.05,0.50,5.00,20.00 μg/L) of TNF-α for 48 hours,and CysLT1 mRNA in 16HBE cells was measured by reverse transcription(RT)-PCR.CysLT1 expression was detected by immunohistochemistry.Results 16HBE cells did not express CysLT1,after the cells were treated with TNF-α,obvious expression of CysLT1 were detected by immunohistochemistry.The weak CysLT1 mRNA expression was observed by RT-PCR in 16HBE cells,and after the cells were treated with TNF-α for 48 hours,CysLT1 mRNA expression were upregulated.When the concentrations of TNF-α were 0.00,0.05,0.50,5.00,and 20.00 μg/L respectively,the relative intensities of CysLT1 mRNA/β-actin were 0.048,0.105,0.177,0.182,0.495,respectively.Conclusions TNF-α can upregulate CysLT1 mRNA expression in 16HBE ceils in a dose-dependent manner.When infected by virus,respiratory tract produces abundant TNF-α.The TNF-α can upregulate the expression of CysLT1 in epithelial cells,enhance inflammation reaction in respiratory tract.This may explain partially the mechanism of exacerbation of asthma induced by respiratory tract infection.

Hematologic parameters and genotype analysis in 166 children with HbH disease in the North Guangxi region / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the characteristics of genotype spectrum and hematologic parameters in children with HbH disease in the North Guangxi region.</p><p><b>METHODS</b>HbH disease was identified by clinical manifestations, routine blood tests and hemoglobin electrophoresis in 166 children who came form the North Guangxi region. Genotypes were determined by Multi-PCR combined with PCR reverse dot blot. DNA sequencing was used when the genotype could not be identified by regular methods.</p><p><b>RESULTS</b>Of the 166 children with HbH disease, 8 genotypes were identified: --SEA/-α3.7 (82 cases), --SEA/-α4.2 (40 cases), --SEA/αCSα (38 cases), --SEA/αQSα (1 case), --SEA/αWSα (1 case), --SEA/αCD43/44 (-C) α (1 case), --SEA/-α3.7 plus CD17 (A→T) (1 case) and --SEA/-α4.2 plus CD41-42(-TTCT) (1 case). One case was confirmed as the heterozygote of --SEA and an unknown mutation. In the 134 cases with complete medical data, 2 had normal hemoglobin levels, 36 manifested mild anemia, 90 manifested moderate anemia, and 6 (genotype: --SEA/αCSα) showed severe anemia because of the coexistence of infection. Children with the genotype of --SEA/-α3.7 (69 cases), --SEA/-α4.2 (31 cases) and --SEA/αCSα (34 cases) had hemoglobin levels of 62-120, 69-127 and 34-110 g/L respectively. The hemoglobin level in the --SEA/αCSα group was significantly lower than in the deletional HbH disease group (genotypes: --SEA/-α3.7 and --SEA/-α4.2 ) (P<0.05). In contrast, MCV levels in the --SEA/αCSα group were significantly higher than in the deletional HbH disease group (P<0.05).</p><p><b>CONCLUSIONS</b>The genotype spectrum of HbH disease is diverse in the North Guangxi region. Deletional genotype is prevalent. The disease is heterogeneous. The children with --SEA/αCSα HbH disease have severer anemia and higher MCV levels than those with deletional HbH disease.</p>

Chest X-ray findings in children with enterovirus 71 infection / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the characteristics of the chest X-ray images in children infected with enterovirus 71.</p><p><b>METHODS</b>A total of 120 children with enterovirus 71 infection between April, 2010 and July, 2011 were classified into three groups according to the disease condition: mild (31 cases), severe (43 cases) and life-threatening (46 cases). The period from the onset of clinical symptoms to the first chest X-ray imaging examination and the results of the first chest X-ray findings were compared among the three groups.</p><p><b>RESULTS</b>The period from the onset of clinical symptoms to the first chest X-ray imaging examination in the mild, severe and life-threatening groups was 26-48 hrs (median 37 hrs), 10-36 h (median 23 hrs) and 2-36 hrs (median 19 hrs) respectively. Chest X-ray abnormalities were initially observed at 30 hrs after the onset of clinical symptoms in the mild group, at 23 hrs in the severe group and at 2 hrs in the life-threatening group (P<0.01). The mild group presented an initial imaging abnormality rate of 5.8%, the severe group 81.3% and the life-threatening group 100%. The life-threatening group showed a significantly higher initial X-ray abnormality rate than the other two groups (P<0.01). In terms of chest X-ray performance, the mild group usually presented lung marking thickening or vagueness. Most children in the severe group presented lung effusion and consolidation. Signs of pulmonary edema were found in the life-threatening group, and lesions in the life-threatening group were characterized by wide distribution and many lung lobe involvements.</p><p><b>CONCLUSIONS</b>The interval between the onset of clinical symptoms and the initial chest X-ray examination, the period of time of, and the onset of clinical symptoms, at which chest X-ray abnormalities, the abnormality rate and the severity of chest X-ray findings may be paralleled to the clinical situation in children with enterovirus 71 infection.</p>

Effect of Iron Deficiency on Hemoglobin A2 Level in Patients with β-Thalassemia / 实用儿科临床杂志

Objective To determine the effect of iron deficiency on hemoglobin A2(HbA2) expression in patients with β-thalassemia.Methods The participants were recruited from the out-patient clinics of the Pediatrics Department and Obstetrics Department of Affiliated Hospital of Guilin Medical College and from some β-thalassemia major families.Blood samples from the participants were used for blood smear tests and hemoglobin electrophoresis and to analyze serum ferritin (SF),3 alpha-globin gene deletions,and 17 beta-globin point mutations.Results Of the 408 individuals,304 were assigned to group A (normal controls),26 to group B (iron deficiency),56 to group C (β-thalassemia),and 22 to group D (β-thalassemia combined with iron deficiency). The results for the comparison of the mean HbA2 values among pairs of groups were as follows: group A vs group B,q=5.074 7,P<0.05; group A vs group C,q=37.650 8,P<0.05; group A vs group D,q=16.043 0,P<0.05;group C vs group D,q=7.682 9,P<0.05; Group B vs group D,q=15.806 6,P<0.05. There were no significant correlation between SF and HbA2 in all 4 groups.Conclusions Iron deficiency decreased the HbA2 level in both controls and individuals with β-thalassemia. HbA2 levels decreased significantly in individuals with both β-thalassemia and iron deficiency as compared with β-thalassemia group alone. However,they remained significantly higher than both the control and iron-deficient groups. Therefore,the elevation of HbA2 could be used to diagnose β-thalassemia reliably even in the presence of iron deficiency.

A study on the (CA)n in FVIII gene in Han ethnic group in Guangxi Zhuang Autonomous Region by amplification polymorphisms combined with silver staining / 中华儿科杂志

<p><b>OBJECTIVE</b>Hemophilia A is an inherited bleeding disorder caused by defects in factor VIII (FVIII) gene. In the present study, the frequencies of the microsatellite alleles at introns 13 and 22 in the factor VIII gene were analyzed in the group of Han nationality in Guangxi Zhuang Autonomous Region to explore their diagnostic value for hemophilia A. These two sites were also used to detect the carriers in 13 hemophilia A families.</p><p><b>METHODS</b>Ninty-one individuals of Han ethnic group in Guangxi Zhuang Autonomous Region (135 X chromosomes) and 13 HA families were subjected to molecular studies. First, these two fragments were PCR amplified simultaneously. Then, silver staining was used later to show their polymorphisms. The investigators selected one sample at random to obtain its lengths of the PCR products at these two sites by ABI310 PCR amplifier. After counting its repeated numbers of (CA) according to the documents concerned, the repeated numbers of the other samples could be counted easily.</p><p><b>RESULTS</b>In the 91 individuals, 6 and 4 alleles were detected at these two sites, respectively. At intron 13 the allele frequencies ranged from 0.0002 to 0.5408 and polymorphism information content (PIC) was 0.5899. At intron 22 the allele frequencies ranged from 0.0444 to 0.4963 and its PIC was 0.5359. The actual heterozygosity for intron 13 and intron 22 were 0.6364 (28/44) and 0.5227 (23/44), respectively. In 13 hemophilia A families with positive history, 9 of them were diagnosed by this method and the diagnosis rate was 69%.</p><p><b>CONCLUSION</b>With high PICs, (CA)n at intron 13 and intron 22 were two valuable sites in the diagnosis of hemophilia A in the population of Han ethnic group in Guangxi Zhuang Autonomous Region. Compared with some other HA restrictive fragment length polymorphisms (RFLP), intron 22 (GT)n (AG)n was more informative.</p>

Influence of Pranlukast on Expression of Eotaxin-3 in Bronchial Epithelia Cells / 实用儿科临床杂志

Objective To investigate whether leukotriene D4 (LTD4) regulates eotaxin-3 (Eot-3) expression in bronchial epithelial cells, and study effect of pranlukst on the regulation.Methods BEAS-2B cells and normal human bronchial epithelia cells were pre- treated with LTD4 for 1 hour,stimulated with interleukin-4, the cells were incubated for 24 hours. Eot-3 protein in supernatant were measured by enzyme linked immunosorbent assay(ELISA). The cells were pretreated with pranlukast in different concentration, then the above procedure was repeated. Results The untreated bronchial epithelial cell expressed Eot-3 protein on a very low level. After stimulating with IL-4 and incubating for 24 hours, Eot-3 production increased significantly. Pretreating the cells with LTD4 enhanced the inducing effect of IL-4. Pranlukast inverted the upregulation of LTD4. Conclusions Upregulating the expression of Eot-3 induced by IL-4 on bronchial epithelial cells may explain partially the mechanism of leukotrienes involving airway allergic inflammation of asthma. The invertion impact on upregulation of LTD4 by pranlukast may be one of mechanisms that leukotrienes receptor antagonist cure asthma.