Compound heterozygous mutations of Fanc A gene in two children with Fanconi anemia / 中华实用儿科临床杂志

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.

Influence of HMGB1/MAPK/m-TOR signaling pathway on cell autophagy and chemotherapy resistance in K562 cells / 中南大学学报(医学版)

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.

Effects of parental anxiety and depression on the feelings and self-concept of children with leukemia / 肿瘤研究与临床

Objective To evaluate the anxiety and depression problems in children parents with leukemia, as well as the problems' influence to the psychosocial characteristics of leukemia children.Methods Twenty long-term survivors of childhood leukemia (group A), thirty children newly diagnosed as leukemia (group B) and fifty age-matched healthy controls (group C) completed the questionnaires allowing assessment of symptoms associated with anxiety, depression and self-concept. At the same time, the anxiety and depression in the parents of children with leukemia were also measured using SAS and SDS. Results The anxiety and depression scores of the parents in study group (48.56±9.23, 51.86±9.53) were much higher than that in normal population (37.23±12.59, 41.88±10.57) (P = 0.000, 0.000, respectively), the positive rate of anxiety and depression symptoms among group B was significantly higher than that among group A (60.0 % vs 25.0 %, 46.7 % vs 20.0 %; P <0.05, respectively). There was a significant positive relation between the depression and anxiety scores in the parents of children with leukemia(r =0.947, P =0.0000). Group A scored significantly higher on subscales of somatization/panic (6.11 ±4.36), generalized anxiety (5.72±4.56),social phobia (7.67±4.19) and the total scale (25.8±13.98) than group C (the score was 3.68±3.39, 2.54±2.99,4.24±2.88 and 15.9±10.52, respectively) (P<0.05, respectively), group B scored significantly higher on subscales of social phobia (6.03 ±2.16) than group C (4.24±2.88) (P =0.016). There was a significantly positive relation between the depression score in children with leukemia and the anxiety (r = 0.309, P = 0.029) & depression(r = 0.342, P = 0.015) scores in their parents. Group A scored significantly higher on the total score of self-concept(60.8±6.25) as well as the subscales of happiness(7.95±1.32) than group G (64.48±7.89 vs 8.64±1.19) (P =0.039, 0.026, respectively); and group B scored significantly higher on subscales of behavior (12.47±1.25), intelligence(10.80±2.12), physical appearance and attributes (8.40±2.66), anxiety (9.93±1.29) and the total scale (59.83±5.87) than group C (14.00±2.17, 12.60±2.96, 9.64±2.30, 11.38+2.18, 64.48±7.89) (P <0.05, respectively). There was a significantly positive relation between the score of gregarization subscale in leukemic children and the anxiety score in their parents (r = 0.337, P = 0.017). Conclusion The findings of our studies have suggested that the parents of children with leukemia are at risk for psychological difficulties, and which have a great influence on the psychological health of their children.

Effects of HLA disparity of two umbilical cord blood units on human engraftment in SCID mice / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the feasibility and characteristics of human engraftment in HLA disparate cord blood transplantation.</p><p><b>METHODS</b>Two human HLA-haploidentical or HLA-mismatched cord blood units were transplanted into sublethally irradiated severe combined immunodeficiency (SCID) mice. The characteristics of engraftment, hematopoietic and immunological reconstitution between the two groups were compared.</p><p><b>RESULTS</b>Two mixed cord blood units can engraft in SCID mice with donor-recipient chimerism and reconstitute hematopoiesis and immunological functions. No unfavorable factors had been observed. Only one of the two cord blood units which had higher colony forming ability in vitro could engraft in most SCID mice as shown by HLA-DQB(1) gene detection. Two HLA-haploidentical cord blood units were simultaneously engrafted in 3 SCID mice.</p><p><b>CONCLUSION</b>Double HLA-haploidentical or HLA-mismatched cord blood can engraft in SCID mice and reconstitute hematopoietic and immunological functions. HLA disparity has no significant effect on survival and engrafting rate. However, in less HLA disparity group, two cord blood units were prone to engraft simultaneously.</p>