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The clinical features and laboratory tests results of two cases with Fanconi anemia (FA) who were admitted to the Department of Children′s Hematology and Endocrinology, the Provincial Hospital Affiliated to Shandong First Medical University in 2017 were analyzed.Sanger sequencing and multiplex ligation-dependent probe amplification(MLPA) of FA-related genes was carried out.One case was female, 4 years and 3 months old.The other case was a 6-year-old male.The main manifestations were recurrent fever, asthenia and bleeding points in both legs.The girl had milk coffee spots scattered on her legs and waist, and her left thumb nail was absent.The boy had no obvious physical examination abnormality, but his left atrium and left ventricle were large and segmental myocardial damage could be seen by echocardiography.Bone marrow biopsies of both cases showed hypo-proliferation (40%) or extremely low proliferation (10%), and no megakaryocyte was found.There were no significant abnormalities in chromosome aberration, single cell gel electrophoresis, cluster of differentiation(CD) 41, CD 55, and CD 59 and chromosome karyotype.Gene sequencing revealed that the two children had compound heterozygous mutations of Fanc A gene, which came from parents.The heterozygous mutation of c1838delT was found in the exon 21 of the female child and her father, which resulted in amino acid shift mutation pIi613Tfs*27.The heterozygous deletion mutations in exons 1-3 of Fanc A gene were found in the female child and her mother by the MLPA results.The gene sequencing analysis of the male child and his family members showed the heterozygous mutation of c4124_4125del in the exon 41 of the child and his mother, which resulted in amino acid shift mutation p. T1375Sfs*49.The heterozygous deletion mutations were observed in exons 23-40 of the male child and his father, according to the MLPA results.The main basis of diagnosis of FA is to sequence the related genes of suspected children.The c1838delT is a new mutation of Fanc A gene.
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Objective To investigate the relationship between the expression of GATA6 mRNA and the occu-rrence of congenital heart disease(CHD). Methods A total of 60 cases of CHD (CHD group)were diagnosed at Obstetrical Department,Rizhao City Maternal and Child Health Hospital,and 60 cases of normal pregnancy women (control group)were collected in March 2013 to May 2017. The expression levels of mRNA of GATA6 in the peripheral blood of pregnant women in 2 groups were detected by using real-time quantitative PCR (qRT-PCR)technique,and the relevant information of 2 groups of mothers before and during pregnancy was collected so as to investigate the rela-tionship between mRNA and GATA6 in CHD. Results There were no significant differences in age,body mass index (BMI),passive smoking,alcohol consumption,pregnancy medication,pregnancy-induced hypertension,high -density lipoproteincholesterol (HDL-C),total cholesterol (TC),triacylglycerol (TG)between pregnant women in the congenital heart disease group and the control group (all P > 0. 05). The CHD group showed a statistical difference significance in the rate of taking folic acid(81. 67％)and gestational diabetes incidence(8. 33％),serum level of LDL-C[(3. 59 ± 0. 46)mmol/ L]compared with the healthy control group [96. 67％,0,(3. 20 ± 0. 44)mmol/ L] (all P < 0. 05). The expression level of GATA6 mRNA in peripheral blood of patients with congenital heart disease (0. 014 ± 0. 005)was significantly lower than that of the control group (0. 129 ± 0. 031),and the difference was statis-tically significant (t = 28. 386,P = 0. 000). Logistic regression analysis showed that the level of in GATA6 mRNA ex-pression increased in the peripheral blood and folic acid intake were protective factors for CHD (all P < 0. 05)during pregnancy;while fetal gestational diabetes,elevated levels of LDL-C are risk factors for congenital heart disease (all P < 0. 05)fetus. Conclusion Down regulation of GATA6 mRNA expression in the maternal peripheral blood is one of the independent risk factors for fetal congenital heart disease.
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To observe the effect of high-mobility group box 1 (HMGB1) on autophagy and chemotherapy resistance in human leukemiacell line (K562) cells, and to explore the underlying mechanisms. Methods: The K562 cells were cultured in vitro and divided into 6 groups: a chemotherapeutic group, a chemotherapeutic control group, a HMGB1 preconditioning group, a HMGB1 preconditioning control group, a HMGB1 siRNA group and a siRNA control group. The chemotherapeutic group was further divided into a vincristine (VCR) group, an etoposide (VP-16) group, a cytosine arabinoside (Ara-C) group, a adriamycin (ADM) group and a arsenic trioxide (As2O3) group. The cell activity was evaluated by cell counting kit-8. The protein levels of HMGB1, microtubule-associate protein1light chain3 (LC3), AMP-activated protein kinase (AMPK) and mammalian target of rapamycin (m-TOR) were determined by Western blotting. The level of serum HMGB1 was evaluated by enzyme-linked immunosorbent assay (ELISA). The autophagy was examined by monodansylcadaverine staining and observed under transmission electron microscopy. Results: Compared with the control group, the cell activity was significantly decreased and the level of serum HMGB1 was significantly increased in the chemotherapeutic (VCR, VP-16, Ara-C, ADM and As2O3) groups (all P<0.05). Compared with the control group, the cell activity and the level of serum HMGB1 were significantly increased in the HMGB1 preconditioning group (both P<0.05). Compared with the siRNA control group, the cell activity and the level of serum HMGB1 were significantly decreased in the HMGB1 siRNA group (both P<0.05). Compared with the control group, the expression of LC3-II and the formation of autophagic bodies were increased in the HMGB1 preconditioning group (both P<0.05), the p-AMPK expression was increased and p-mTOR expression was decreased (both P<0.05). Conclusion: HMGB1 can increase the autophagy and promote chemotherapy resistance through the pathway of AMPK/m-TOR in K562 cells.