Analysis of Thalassemia Gene Carriers in 19 482 Pregnant Women from 2015 to 2019 in Guilin, Guangxi / 中国实验血液学杂志

OBJECTIVE@#To investigate the types and frequencies of thalassemia genes carried by the pregnant women in Guilin, Guangxi Zhuang Autonomous Region, China.@*METHODS@#From January 2015 to December 2019, blood samples of the pregnant women who visited the Outpatients of Obstetrics clinic and Eugenics Genetic clinic in Affiliated Hospital of Guilin Medical University were collected. Gap-PCR was used to detect deletional α-thalassemia, PCR-RDB to detect the gene mutations of non-deletional α-thalassemia and β-thalassemia, and MLPA or DNA sequencing to detect rare thalassemia mutations. Different types and frequencies of thalassemia mutations carried by pregnant women were analyzed statistically.@*RESULTS@#A total of 19 482 blood samples were collected, including 3 801 thalassemia gene carriers (19.51%). Seven types of α-thalassemia gene mutation were detected with a carrier rate of 15.43%. Among them, --@*CONCLUSION@#Guilin is a high-risk area for thalassemia. Alpha-thalassemia is dominated by --

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.

Paternally originated Wolf-Hirschhorn syndrome detected by multiplex ligation-dependent probe amplification and microarray comparative genomic hybridization / 中华儿科杂志

<p><b>OBJECTIVE</b>To confirm the diagnosis of a Wolf-Hirschhorn syndrome by family study using both cytogenetic and molecular genetic techniques.</p><p><b>METHOD</b>G-band karyotyping was performed for all the 6 members in the family. Multiplex ligation-dependent probe amplification (MLPA) was used to detect the chromosome abnormality for the proband, his father and brother. Microarray comparative genomic hybridization (Array-CGH) was carried out to map the exact chromosomal breakpoints for the proband.</p><p><b>RESULT</b>The proband presented with a typical face, delayed growth and hypotonia in Wolf-Hirschhorn syndrome. His G-band karyotype was 46, XY, der(4)t(4;8) (p16.2; p23.1)pat. MLPA showed 4pter loss and 8pter gain. Array-CGH revealed an XY male with a 3.781 Mb deletion of 4p16.3-p16.2 and a 6.760 Mb duplication of 8p23.3-p23.1. The proband's brother has mental retardation and skeletal abnormalities. His G-band karyotype was 46, XY, der(8)t(4;8)(p16.2;p23.1)pat. MLPA showed 4pter gain and 8pter loss. The proband's father had normal phenotype with a balanced translocation of 46, XY, t(4;8)(p16.2;p23.1)pat. MLPA showed a normal result. The proband's grandfather showed a normal phenotype with a balanced translocation 46, XY, t(4;8)(p16.2;p23.1). The other members in the family showed normal phenotypes with normal karyotypes.</p><p><b>CONCLUSION</b>The proband has features of Wolf-Hirschhorn syndrome with partial monosomy 4p and partial trisomy 8p. The proband's brother has a partial trisomy 4p and partial monosomy 8p. The derived chromosomes are inherited from paternal balanced translocation t(4;8)(p16.2;p23.1).</p>

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>

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.