AIM: To investigate the antiplatelet aggregation of naringenin and its possible mechanism . METHODS:The effects of naringenin at different concentrations on adenosine diphosphate ( ADP)-induced platelet aggre-gation and platelet spreading on immobilized fibrinogen were detected by aggregational detector and observed under fluores -cence microscope, respectively.The expression of phosphoinositide 3-kinase (PI3K) and the phosphorylation of Akt were analyzed by Western blot .RESULTS:Naringenin significantly inhibited ADP-induced platelet aggregation in a dose-de-pendent manner in vitro and in vivo, and inhibited platelet ' s spreading on immobilized fibrinogen in vitro.Further studies indicated that naringenin inhibited platelet activation accompanied by attenuating not only the activation of PI 3K, but also the phosphorylation of Akt .Moreover, naringenin combined with LY294002 additively inhibited the ADP-induced platelet aggregation.CONCLUSION:Naringenin may inhibit platelet activation by regulation of PI 3K/Akt signaling pathway.

AIM: To investigate effect of naringenin on ADP-induced platelet aggregation and its possible mechanism.METHODS: The levels of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) were measured in the platelets with ADP stimulation using ELISA in the presence or absence of different concentrations of naringenin.The effect of naringenin at different concentrations on the change of phosphodiesterase (PDE) activity was measured by high efficiency liquid chromatography.The effects of naringenin at different concentrations on phosphorylation of vasodilator-stimulated phosphoprotein (VASP) at positions Ser157 and Ser239 in washed platelets with ADP stimulation were analyzed by Western blot.The phosphorylation of VASP at Ser239 was also analyzed in the presence of protein kinase A (PKA), protein kinase G (PKG), or protein kinase C (PKC) inhibitors before incubation with naringenin.The platelet aggregation was measured in the presence of PKA or PKG inhibitors before incubation with naringenin.RESULTS: Naringenin elevated cGMP levels significantly but not cAMP levels in the platelets with ADP stimulation in a dose-dependent manner.Naringenin inhibited PDE activity.Naringenin increased the phosphorylation of VASP at Ser239 in a dose-dependent manner in the platelets with ADP stimulation but only modest changes in the phosphorylation at position Ser157.The phosphorylation level of VASP at Ser239 position was inhibited when the platelets were treated with PKA inhibitor before incubation with naringenin.Incubation of platelets with neither PKG nor PKC inhibitors before treatment with naringenin affect the phosphorylation of VASP at Ser239.Pretreatment with PKA inhibitor but not PKG inhibitor significantly reversed the antiplatelet aggregation by naringenin in ADP-stimulated platelets.CONCLUSION: Naringenin may inhibit platelet activation through the elevation of cGMP-and PKA-mediated VASP phosphorylation.

Humans ; Muscular Dystrophy, Duchenne ; physiopathology ; therapy ; Respiratory Physiological Phenomena

OBJECTIVETo analyze clinical characteristics of a combination of dystrophinopathies and Klinefelter's syndrome (karyotype 47, XXY) in one patient.

METHODThe patient was diagnosed as Duchenne muscular dystrophy (DMD) and Klinefelter's syndrome in Beijing Children's Hospital in March, 2013. The clinical manifestations, physical examinations and laboratory test results were analyzed respectively. The clinical characteristics of four cases reported previously were analyzed as well.

RESULTThe 8.5 years old boy presented with symptoms of walking disorder and developmental delay. The patient had facial dysmorphism, waddling gait, Gower's manoeuvre and enlarged calves.Serum creatine kinase level was 21 040 U/L, and he had mild intellectual impairment. Deletions of exons 49-54 of the dystrophin gene were found.Gene dosage analysis revealed a heterozygous deletion in his mother. Five cases have been reported till now, their age ranged from 3.5 to 18 years; 3 of them were DMD, while the other 2 cases were Becker muscular dystrophy (BMD). One of them, detected in pedigree study, whose weakness was minimal in contrast to the proband. The others came to the hospital because of walking disorder or developmental delay. All the patients had enlarged calves, some of them also had Gower's manoeuvre and waddling gait. The patients' height was between 3 rd and 50 th percentile, while 2 of them had facial dysmorphism.Some degree of mental impairment is usual. Their serum creatine kinase were 2 469-24 750 U/L.One of them was detected in pedigree study. Three of them were diagnosed by muscle biopsy, while in the other one mutation analysis was used.

CONCLUSIONThe combination of dystrophinopathies and Klinefelter's syndrome is quite rare, and has clinical features of these two diseases. Mutation analysis (or muscle biopsy) and karyotype analysis can finally diagnose the syndrome.

Child ; Creatine Kinase ; blood ; DNA Mutational Analysis ; Dystrophin ; genetics ; metabolism ; Exons ; genetics ; Gene Deletion ; Heterozygote ; Humans ; Intellectual Disability ; Klinefelter Syndrome ; complications ; diagnosis ; genetics ; Male ; Muscle Weakness ; etiology ; Muscular Dystrophy, Duchenne ; complications ; diagnosis ; genetics ; Mutation ; Pedigree

Objective To investigate the effects of skin/muscle incision and retraction(SMIR)on mechanical paw withdrawal threshold and the ability of spatial learning and memory in immature rats after adulthood. Methods 27 male SD rats aged 3 weeks and weighing 60 ~ 80 g were randomly divided into 3 groups(n = 9):control group(group C),sham operation group(group Sham)and skin/muscle incision and retraction group (group SMIR). Group SMIR received operation for skin/muscle incision and retraction. Sham group received skin/muscle incision but no retraction.No surgery was operated on C group. Pain behavior was assessed by mechanical paw withdrawal threshold(MWT)to von Frey filament stimulation before and 1,3,7,12,22 and 32 days after operation.The effects of spatial learning and memory function were assessed by Morris water-maze test at 33 days after operation. Results Mechanical paw withdrawal threshold of group SMIR decreased 1 day after operation (P<0.05)and showed no significant difference before and 3,7,12,22,32 days after operation in 3 groups(P >0.05). In Morris water-maze test,compared with Sham and C group,the average escape latency in SMIR was sig-nificantly longer in the water maze navigation experiment(P < 0.01);the ratios of time and path in the quadrant of the platform were obviously lower in SMIR(P < 0.01). There was no statistical difference between sham and C group(P>0.05).Conclusion SMIR did not cause chronic pain but may cause a decrease in the ability of spatial learning and memory in immature rats.

AIM: Previous studies have suggested that big stick design (BSD) method can only be used in clinical trials of two treatments with equal proportion, which has good statistical performance and has become the recommended choice of randomized methods. This study expands BSD method, so that it can be applied to three groups, and provides more randomized methods for clinical trials. METHODS: On the basis of BSD method used in two treatments with equal proportion, the derivation conditional allocation probability of BSD method used in three treatments with equal proportion was carried out. BSD method was compared with simple randomization (SR) method, permuted block design (PBD) method and block urn design (BUD) method by Monte-Carlo simulation in balance and randomness. RESULTS: In terms of balance, PBD method was the best, followed by BUD method, BSD method, and SR method was the worst. In terms of randomness, SR method was the best, followed by BSD method, BUD method and PBD method. The comprehensive performance showed that BSD method was better than BUD method, PBD method and SR method. CONCLUSION: The expanded BSD method used in three treatments with equal proportion has good comprehensive performance, and it can be the recommended randomization method for clinical trials of three treatments with equal proportion.