Detecting rare thalassemia in children with anemia using third-generation sequencing.

BACKGROUND: In China, conventional genetic testing methods can only detect common thalassemia variants. Accurate detection of rare thalassemia is crucial for clinical diagnosis, especially for children that need long-term blood transfusion. This study aims to explore the application value of third-generation sequencing (TGS) in the diagnosis of rare thalassemia in children with anemia. METHODS: We enrolled 20 children with anemia, excluding from iron deficiency anemia (IDA). TGS was employed to identify both known and novel thalassemia genotypes, while sanger sequencing was used to confirm the novel mutation detected. RESULTS: Among the 20 samples, we identified 5 cases of rare thalassemia. These included ß-4.9 (hg38,Chr11:5226187-5231089) at HBB gene, α-91(HBA2:c.*91delT), αCD30(HBA2:c.91-93delGAG), Chinese Gγ+(Aγδß)0(NG_000007.3: g .48795-127698 del 78904) and delta - 77(T > C)(HBD:c.-127T>C). Notably, the -SEA/α-91α genotype associated with severe non-deletional hemoglobin H disease (HbH disease) has not been previously reported. Patients with genotypes ß654/ß-4.9 and -SEA/α-91α necessitate long-term blood transfusions, and those with the -SEA/αCD30α, Chinese Gγ+(Aγδß)0 and delta thalassemia demonstrate mild anemia. CONCLUSIONS: TGS demonstrates promising potential as a diagnostic tool for suspected cases of rare thalassemia in children, especially those suspected to have transfusion-dependent thalassemia (TDT).

Suppression of MIF-induced neuronal apoptosis may underlie the therapeutic effects of effective components of Fufang Danshen in the treatment of Alzheimer's disease.

Fufang Danshen (FFDS or Compound Danshen) consists of three Chinese herbs Danshen (Salviae miltiorrhizae radix et rhizome), Sanqi (Notoginseng radix et rhizome) and Tianranbingpian (Borneolum, or D-borneol), which has been show to significantly improve the function of the nervous system and brain metabolism. In this study we explored the possible mechanisms underlying the therapeutic effects of the combination of the effective components of FFDS (Tan IIA, NG-R1 and Borneol) in the treatment of Alzheimer's disease (AD) based on network pharmacology. We firstly constructed AD-related FFDS component protein interaction networks, and revealed that macrophage migration inhibitory factor (MIF) might regulate neuronal apoptosis through Bad in the progression of AD. Then we investigated the apoptosis-inducing effects of MIF and the impact of the effective components of FFDS in human neuroblastoma SH-SY5Y cells. We observed the characteristics of a "Pendular state" of MIF, where MIF (8 ng/mL) increased the ratio of p-Bad/Bad by activating Akt and the IKKα/ß signaling pathway to assure cell survival, whereas MIF (50 ng/mL) up-regulated the expression of Bad to trigger apoptosis of SH-SY5Y cells. MIF displayed neurotoxicity similar to Aß1-42, which was associated with the MIF-induced increased expression of Bad. Application of the FFDS composite solution significantly decreased the expression levels of Bad, suppressed MIF-induced apoptosis in SH-SY5Y cells. In a D-galactose- and AlCl3-induced AD mouse model, administration of the FFDS composite solution significantly improved the learning and memory, as well as neuronal morphology, and decreased the serum levels of INF-γ. Therefore, the FFDS composite solution exerts neuroprotective effects through down-regulating the level of Bad stimulated by MIF.

[Effect of calcineurin Aalpha gene overexpression on the myocardium apoptosis induced by hypoxia-reoxygenation and adrenergic receptors].

OBJECTIVE: To investigate the effect of calcineurin AalphacDNA (AdCnAalpha) overexpression as a result of adenovirally mediated gene transfer on neonatal rat cardiac myocyte apoptosis induced by hypoxia-reoxygenation (H/R) and adrenergic receptors. METHODS: Neonatal rat cardiac myocytes were cultured for 20 h after AdCnAalpha transfection, and treated with isoproterenol (10 micromol/L) and 24 h of hypoxia followed by 4 h of reoxygenation (24H/4R). The cardiac myocyte apoptosis induced by the treatments was assessed by flow cytometry and DNA laddering, and the levels of calcineurin, p38 and phosphorylation p38 (p-p38) were determined by Western blotting and (or) RT-PCR. RESULTS: AdCnAalpha transfection promoted cultured neonatal rat cardiac myocyte apoptosis induced by isoproterenol+24H/4R as compared with the treated cells without transfection (14.247-/+0.525 vs 10.763-/+1.554, P<0.01), along with greater phosphorylation p38 protein expression (1.60-/+0.22 vs 2.42-/+0.19, P<0.01). The levels of p38 underwent no obvious change after AdCnAalpha transfection (P<0.05). CONCLUSIONS: AdCnAalpha transfection can promote cardiac myocyte apoptosis induced by H/R and adrenergic receptors, the mechanism of which might be associated with p38 mitongen-activated protein kinase (p38MAPK) activation.