[Analysis of the gene mutation in an inherited FVII deficiency patient].

OBJECTIVE: The identify the gene defect of an inherited FVII deficiency patient. METHODS: The promoter, all the exons and exon-intron boundaries and 3' UTR of F7 gene of the proband were analyzed by direct sequencing. The defected mutations were confirmed by sequencing the complementary strand. The mutations would be screened in the related database and 150 healthy donors to identify the SNP. By splice site prediction, we analyzed the pathogenesis of defected mutations. RESULTS: Genetic analysis revealed G to A transition at 15975 in the intron 6 of F7 gene (IVS6-1G>A) and A to G transition at 16813 in the intron 7 of F7 gene (IVS7+7 A>G). According to the fruitfly, the acceptor site could not be recognized when a G to A substitution took place. The closest candidate splice site was located 132 bp downstream. The distance was probably too far to allow the use of the cryptic splice site and resulted in the skipping of exon6. A to G transition at 16813 in the intron 7 of F7 gene could not change the splice site, but modify a different molecular interaction that is important for the splice process. CONCLUSION: The heterozygous mutation of IVS6-1G>A combined with polymorphism of IVS7+7 A>G in F7 gene relates to the FVII deficiency.

[Sensitivity and specificity of noninvasive prenatal fetal RhD genotesting: a meta-analysis].

OBJECTIVE: To explore the sensitivity, specificity and clinical validity of fetal Rh genotyping from maternal blood. METHODS: A comprehensive literature search of PubMed, Embase and Web of Science was performed for describing fetal RhD determination from maternal blood. The inclusion criteria were established based on the validity criteria for diagnostic research. And the eligible entries were collected and analyzed with MetaDisc4.0. RESULTS: This meta-analysis included 55 studies with a total of 17 138 samples. The random-effect model was used for analysis because of heterogeneity. The pooled sensitivity and specificity were 98.5% and 97.3% respectively. The SROC curve was plotted and the area under the curve (AUC) calculated (AUC = 0.994). The subgroup and sensitivity analyses were performed. The sensitivity of 25 studies with samples<100 (94.6%) was significantly lower than those of 19 studies with samples 100-300 (98.5%) and 11 studies with samples>300 (99.0%) (χ² = 36.800, 106.062, P < 0.05). The sensitivity of 19 studies with samples 100-300 (98.5%) was not different from that of 11 studies with samples >300 (99.0%)( χ² = 3.068, P > 0.05). CONCLUSIONS: Noninvasive prenatal diagnosis of fetal RhD status from maternal blood represents a significant achievement in the application of research with high sensitivity and specificity. It may be applied for screening testing of all RhD⁻ negative pregnant women.

MnTBAP increases BMPR-II expression in endothelial cells and attenuates vascular inflammation.

AIMS: The endothelium plays an important role during vascular inflammation. Previous data have demonstrated a high expression level of manganese-superoxide dismutase (MnSOD) in endothelial cells and suggested an important role of MnSOD in several cardiovascular diseases. Manganese (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) has been shown to mimic some of the effects of MnSOD and prevented the development of diabetes and obesity. However, its effect on vascular inflammation and the underlying mechanism is still unknown. METHODS AND RESULTS: Leukocyte adhesion was evaluated in-vivo and in-vitro using dynamic flow chamber and intravital microscopy in mice. Expression of adhesion molecules induced by TNFα and adhesion of leukocytes to the vessel wall were inhibited by MnTBAP. The anti-inflammatory effect of MnTBAP was partly mediated by up-regulation of the BMPR-II and Smad dependent pathway. Additionally, MnTBAP decelerated the turn-over of endogenous BMPR-II. CONCLUSION: Our data demonstrate that MnTBAP activates Smad signaling, preserves the turn-over of BMPR-II and elicits anti-inflammatory effects in endothelial cells, partly mediated by BMPR-II. This finding suggests a potential therapeutic impact of MnTBAP in the treatment of vascular inflammation.

Puerarin attenuates smoke inhalation injury by regulation of Th1/Th2 expression and inhibition of Th17 cells in rats.

OBJECTIVE: Puerarin, a kind of traditional Chinese medicine, possesses immunomodulatory property. However, the immunomodulatory effects of puerarin on smoke inhalation injury have not been determined. The aim of the current study was to investigate the therapeutic efficacy of puerarin on gunpowder smog-induced acute lung injury in rats via regulation of Th1/Th2/Th17 expression. MATERIALS AND METHODS: Wistar rats were equally randomized to four groups (normal control group, puerarin control group, smoke inhalation injury group, puerarin treatment plus smoke inhalation injury group). The severity of lung injury was evaluated by histopathology, myeloperoxidase (MPO) activity in lung homogenates, cell counting in bronchoalveolar lavage fluid (BALF), and lung vascular permeability parameters including lung wet to dry weight ratio and protein concentration in BALF. Flow cytometry was used to analyze the expression of Th1/Th2/Th17 lymphocytes in blood of rats. RESULTS: Puerarin showed significant therapeutic effects against neutrophil infiltration and tissue injury, as evidenced by histopathological findings and MPO activity. Lung vascular permeability was also relieved by puerarin administration. Additionally, puerarin significantly decreased the number of neutrophils and lymphocytes in BALF compared with smoke inhalation injury group. Furthermore, puerarin increased Th1 immunity and reduced Th2 and Th17 responses and thereby altering the Th1/Th2/Th17 imbalance induced by smoke inhalation. CONCLUSIONS: Our findings suggested that puerarin suppressed inflammatory responses in gunpowder smog-induced acute lung injury by regulation of Th1/Th2/Th17 expression, and may be a potential therapeutic agent for smoke inhalation injury.

Protective effects of edaravone combined puerarin on inhalation lung injury induced by black gunpowder smog.

OBJECTIVE: The present study aimed to investigate the combined effects of puerarin with edaravone on inhalation lung injury induced by black gunpowder smog. MATERIALS AND METHODS: Male Wistar rats were divided into five groups (control group, edaravone group, puerarin group, edaravone combined with puerarin group and inhalation group). The severity of pulmonary injuries was evaluated after inducing acute lung injury. Arterial blood gas, inflammatory cytokines, biochemical, parameters, cell counting, W/D weight ratio and histopathology were analyzed. Results in lung tissues, either edaravone or puerarin treatment alone showed significant protective effects against neutrophil infiltration and tissue injury, as demonstrated by myeloperoxidase activity and histopathological analysis (all p<0.05). In addition, combined treatment with both edaravone and puerarin demonstrated additive protective effects on smog-induced lung injury, compared with single treatment. CONCLUSIONS: Combination of edaravone and puerarin shows promise as a new treatment option for acute lung injury/acute respiratory distress syndrome patients.