G,the noval insertion mutation of c.2298_2299insC is identified in Chinese patients.

Asian Continental Ancestry Group ; Bartter Syndrome ; genetics ; Child ; Chloride Channels ; genetics ; Humans ; Male ; Mutation

OBJECTIVEThis study aimed to investigate the mechanism of primary cortical neuron injury induced by high concentrations of copper by observing the effect of aceticum culture medium on apoptosis of rat primary cortical neurons and expression of cleaved caspase 3, caspase 8 and caspase 9.

METHODSPrimary cortical neurons were cultured for 72 hrs and then exposed to different concentrations of aceticum culture medium (20, 40 and 80 microM). The viability of neurons was detected by the MTT method. Apoptosis was observed by Hoechst33258 and flow cytometry Annexin V/PI. Expression of caspase 3, caspase 8 and caspase 9 was measured by Western blot.

RESULTSFollowing incubation with aceticum culture medium, apoptosis of neurons was induced. Theviability of neurons was remarkably reduced and the rate of apoptosis was tremendously increased in a concentration dependent manner. Caspase 8 and caspase 9 were activated in 20 microM of copper aceticum culture medium 4 hrs after incubation and peaked at 48 hrs in various concentrations of copper aceticum culture medium, presenting with a time and concentration dependent manner. The activated caspase 3 was observed in 20 microM of copper aceticum culture medium 24 hrs after incubation, which was later than the activated caspase 8 and caspase 9. Caspase 3 expression reached a peak 48 hrs in various concentrations of copper aceticum culture medium, presenting with a time and concentration dependent manner.

CONCLUSIONSThe apoptosis of primary cortical neurons can be induced by copper. Caspase 3, caspase 8 and caspase 9 cascade reaction may involve in the apoptosis of copper induced rat primary cortical neurons.

Animals ; Apoptosis ; drug effects ; Blotting, Western ; Caspase 3 ; physiology ; Caspase 8 ; physiology ; Caspase 9 ; physiology ; Cell Survival ; drug effects ; Cells, Cultured ; Cerebral Cortex ; drug effects ; Copper ; toxicity ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the association of single nucleotide polymorphisms (SNP) rs22833188 and rs2833195 in TIAM1 gene with the susceptibility to Kawasaki disease (KD) and its clinical characteristic in children.

METHODSA case-control study was performed in this study. One hundred and eighty-eight children with KD and 197 normal children served as controls were enrolled. The genotypes of two SNPs rs22833188 and rs2833195 in TIAM1 gene were detected using PCR-RFLP.

RESULTSThere were no significant differences in the genotype (AA, AG and GG) and allele frequencies of SNP rs2833188 between the KD and control groups. Significant differences in the genotype (CC, GC and GG) frequency of SNP rs2833195 were noted between the KD and control groups (P=0.017). The frequency of C allele in the KD group was higher than in the control group (P=0.015). The polymorphism of SNP rs2833188 was associated with the occurrence of rash (P=0.011), and the polymorphism of SNP rs2833195 was associated with the occurrence of conjunctival hyperemia (P=0.021).

CONCLUSIONSThe polymorphism of rs2833195 in TIAM1 gene is associated with the susceptibility to KD. The polymorphisms rs2833188 and rs2833195 in TIAM1 gene may be associated with some clinical characteristics in children with KD.

Child ; Child, Preschool ; Female ; Genetic Predisposition to Disease ; Genotype ; Guanine Nucleotide Exchange Factors ; genetics ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; genetics ; Polymorphism, Single Nucleotide ; T-Lymphoma Invasion and Metastasis-inducing Protein 1

Objective To investigate copper-induced apoptosis of primary cultured rat cortical neurons and the protective effect of thioredoxin,an apoptosis signal regulated kinase-1(ASK1)inhibitor,and explore the role of ASK1-mediated c-Jun N-terminal kinase(JNK)/caspase-3 signaling pathway in the neurotoxicity of copper.Methods The changes in the viability and apoptosis of primary cultured rat neurons following exposure to cupric acetate and thoredoxin were assessed using MTT assay and flow cytometry.Western blotting was used to detect the expressions of phosphorylated ASK1(p-ASK1),p-JNKand caspase-3 in the exposed cells.Results Administration of cupric acetate in the cell culture resulted in a concentration-dependent increase of cell apoptosis and a reduction of the viability of the neurons,and the effect was antagonized by thoredoxin.The expression of p-ASK1 and p-JNK began to increase in the neurons at 4h after cupric acetate exposure,and reached the peak level at 48h,showing a time-and concentration-dependent pattern of the changes.Activated caspase-3 was expressed at 24h after the exposure,and the peak expression occurred at 48h.The application of thoredoxin significantly lowered the expressions of p-ASK1,p-JNK and caspase-3,and increased the viability and reduced the apoptotic rate in the exposed neurons(P＜0.05).Conclusions Copper can induce apoptosis of primary cultured rat cortical neurons,in which process ASK1-mediated JNK/caspase-3 signaling pathway may play a critical role.Thoredoxin can protect the cortical neurons from injuries induced by the copper.

BACKGROUND: Poly(3-hydroxybutyrate-4-hydroxybutyrate) (P3HB4HB) is a kind of polymer material that can be completely degraded, has good film-forming property and physical properties, but has poor hydrophilicity. OBJECTIVE: To prepare P3HB4HB/polyvinyl alcohol (PVA) coaxial electrospun scaffolds, and to investigate the physical properties and biocompatibility of scaffolds in vitro. METHODS: We prepared P3HB4HB electrospun scaffold, PVA electrospun scaffold and P3HB4HB/PVA coaxial electrospun composite scaffold, and then detected the morphology and characterization, contact angle, and tensile mechanical properties of the scaffolds. Passage 4 bone marrow mesenchymal stem cells (BMSCs) from Sprague-Dawley rats were seeded on the three kinds of scaffolds. Cell adhesion rate was detected at 1, 3, 6 hours after seeding; cell proliferation was detect at 1, 3, 5, 7 days after seeding; and cell viability was observed fluorescence staining at 7 days after seeding. Passage 4 BMSCs were seeded onto the three kinds of scaffolds followed by 14 days of osteogenic and chondrogenic induction. Then, alizarin red staining and toluidine blue staining were used to verify BMSCs differentiation potentials. RESULTS AND CONCLUSION: (1) Scaffold morphology: Under the scanning electron microscope, the structure of the scaffold in each group was a three-dimensional interconnected network. The fiber diameters of P3HB4HB electrospun scaffold and P3HB4HB/PVA electrospun scaffold were homogeneous and ordered. The P3HB4HB/PVA scaffold showed an obvious core-shell structure under the transmission electron microscope. (2) Scaffold characterization: The tensile strength, tensile modulus and maximum stress of the P3HB4HB and P3HB4HB/PVA scaffolds were significantly higher than those of the PVA electrospun scaffold (P < 0.05). The contact angle of the P3HB4HB/PVA composite scaffold was less than 90°. (3) Cell adhesion rate was ranked as follows: PVA electrospun scaffold group >P3HB4HB/PVA composite scaffold group > P3HB4HB electrospun scaffold group (P < 0.05). (4) Proliferation and activity of cells: The cell proliferation of the P3HB4HB/PVA composite scaffold group was faster than that of the other two groups at 5 and 7 days (P < 0.05). There were more viable cells on the PVA electrospun scaffold and composite scaffold than on the P3HB4HB electrospun scaffold. (5) Cell differentiation: Osteogenesis and cartilage specific staining of the composite scaffold were stronger than those in the other two groups. Overall, the P3HB4HB/PVA coaxial electrospun scaffold has good biocompatibility and a certain mechanical strength.

OBJECTIVETo investigate the association between two single nucleotide polymorphisms (SNPs), rs9390754 and rs4840200, in the glutamate receptor 2 (GRIK2) gene and the genetic susceptibility to epilepsy (EP) in the Han population in Central China.

METHODSA case-control study was performed in 284 EP children (including 132 children with refractory epilepsy) and 315 normal children from Central China. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to determine the genotypes of the two SNPs rs9390754 and rs4840200. The genotype frequency was compared between groups.

RESULTSThe frequencies of GG, GA, and AA genotypes of SNP rs9390754 showed a significant difference between the EP and normal control groups (P=0.016). The allele frequency also showed a significant difference between the two groups (P=0.002). The frequencies of CC, CT, and TT genotypes of SNP rs4840200 and allele frequency showed no significant differences between the two groups. The C allele frequency of SNP rs4840200 in the refractory EP subgroup was significantly higher than in the non-refractory EP subgroup (OR=1.435, 95% CI: 1.021-2.016, P=0.037).

CONCLUSIONSIn the Han population in Central China, the polymorphisms of SNP rs9390754 in the GRIK2 gene may be associated with EP susceptibility, and the C allele of SNP rs4840200 may be a genetic risk factor for the development of drug resistance in children with EP.

Child ; Child, Preschool ; Epilepsy ; etiology ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Polymorphism, Single Nucleotide ; Receptors, Kainic Acid ; genetics ; Risk Factors

Objective To investigate whether NADPH oxidase is involved in brain damage of ATP7Btx-J mice through interfering oxidative stress.Methods ATP7Btx-J mice (20 weeks old),wild-type (WT) mice (20 weeks old) and apo-ATP7Btx-J mice (given the NADPH oxidase inhibitor apocynin) were chosen in our study; apo-ATP7Btx-J mice were treated by daily oral administration with 200 mg/kg of apocynin since 16 weeks old till 20 weeks old.Copper concentration was determined by inductively coupled plasma-mass spectrometry (ICP-MS) and NADPH oxidase activity was detected by colorimetric method.The superoxide level was measured using superoxide-sensitive fluorescent probe dihyroethidine (DHE).The protein expression level of Cleaved caspase-3 was analyzed by Western blotting.The level of neuronal apoptosis was assayed with TUNEL method.Results The copper content in the striatum region of ATP7Btx-J mice was significantly higher than that of wild-type (WT) mice (P＜0.05).The activity ofNADPH oxidase and concentration of superoxide anion in the striatum region of ATP7Btx-J mice were significantly higher than those of WT mice (P＜0.05); those in the striatum region of apo-ATP7Btx-J mice were significantly lower than those of ATP7Btx-J mice (P＜0.05).The protein expression of Cleaved caspase-3 and the level of neuronal apoptosis in the striatum region of ATP7Btx-J mice were significantly higher than those of WT mice (P＜0.05); those in the striatum region of apoATP7Btx-J mice were significantly lower than that of ATP7Btx-J mice (P＜0.05).Conclusion NADPH oxidase may play a role in neuronal apoptosis in the striatum region of ATP7Btx-J mice through oxidative stress,and apocynin can protect the nervous system decreasing the NADPH oxidase level.

Objective: To summarize the outcomes of different types of pulmonary atresia in neonates treated by ductus arteriosus stenting. Methods: This study was a retrospective cohort study. A total of 19 neonates who had pulmonary atresia treated by ductus arteriosus stenting in Xinhua Hospital, Shanghai Jiao Tong University School of Medicine from April 2014 to June 2021 were included. They were divided into the intact ventricular septum (PA-IVS) group and the ventricular septal defect (PA-VSD) group. Ductus arteriosus stents were implanted by different approaches. These children were followed up regularly at the 1, 3, 6, and 12 months after the surgery and annually since then to evaluate the outcome. Independent sample t-test was used for the statistical analysis. Results: There were 12 children in PA-IVS group and 7 in PA-VSD group. All of them were full term in fants. The gestational age of the PA-IVS group and the PA-VSD group was (38.8±1.1) and (37.7±1.8) weeks, the birth weights were (3.2±0.4) and (3.4±1.1) kg, and the age at operation was (10±9) and (12±7) days, respectively, without significant difference (all P>0.05). Among the 12 children with PA-IVS, 9 had stents successfully implanted through the femoral artery and 3 through the femoral vein. Of the 7 children with PA-VSD, 2 had the stents successfully implanted via the femoral artery and 2 failed, and the remaining 3 had stents successfully implanted via the left carotid artery. There was no postoperative thromboembolism, arteriovenous fistula, pseudoaneurysm or other vascular complications. Five children with PA-VSD who had successful operations were followed up at 6 months of age. They all had the operation for pulmonary atresia, repair of the ventricular septal defect, removal of arterial duct stents, and ligation of the arterial duct. All children survived without any stent displacement or stenosis and biventricular circulation was achieved during the follow-up. Conclusions: Ductus arteriosous stenting can be the first-stage treatment for children with PA-IVS and PA-VSD. In addition to the traditional femoral vein and femoral artery approach, the carotid artery can be used as a route for stent placement.
Child ; Infant, Newborn ; Humans ; Infant ; Pulmonary Atresia/surgery* ; Ductus Arteriosus ; Retrospective Studies ; China ; Heart Defects, Congenital ; Ductus Arteriosus, Patent/surgery* ; Heart Septal Defects, Ventricular ; Stents

OBJECTIVETo explore demographic characteristics, current diagnosis and treatment patterns of chronic myelogenous leukemia (CML) patients in China.

METHODSData of hospitalized CML patients in 2005 whole year and outpatient information (July 1 through September 30, 2006) from 15 hospitals throughout China were analyzed.

RESULTSA total of 1824 CML cases were analyzed, including 722 inpatients and 1102 outpatients. The male/female ratio was 1.78:1. The median age at diagnosis was 40.02 (2.45 - 83.29) years old, 90.41% of the patients were diagnosed at chronic phase. Proportion of accelerated phase or blast crisis patients increased to 21.66% during study period. 93.20% of the patients received blood routine and bone marrow morphologic examination at diagnosis and in monitoring; 70.29% were performed cytogenetic analysis and 51.54% performed molecular measurement in addition. The most common therapy for CML treatment was hydroxycarbamide. The proportion of patients treated with imatinib and interferon was 37.45% and 25.55%, respectively. Of 722 inpatients, 164 (22.72%) received hemotopoietic stem cell transplantation (HSCT). The proportions of accelerated phase and blast crisis patients treated with imatinib were 48.28% and 48.42%, respectively, being significantly higher than that of chronic phase patients (35.9%) (P < 0.05). The mean imatinib dosage administered in the three phases patients did not differ significantly. Imatinib resistance rates were 6.87% and 16.28% for outpatient and inpatient, respectively. In the outpatient group, the primary resistance to imatinib occurred comparably to the secondary resistance (68.75%), while primary resistance was predominant in inpatient group (65.71%). The intolerance rates of imatinib for outpatient and inpatient were 3.21%, 11.63%, respectively. The majority of patients treated with imatinb were not monitored in time: 63.38% patients evaluated hematologic response after 3 months of treatment, proportions of patients received cytogenetic examination after 6 months and 12 months of treatment were 41.41% and 27.35%, respectively. Mean cost for HSCT was 213 092 +/- 125 890 RMB.

CONCLUSIONSCML in China tends to afflict younger population than in Western countries. Most patients were diagnosed in the chronic phase. Due to restriction of financial support, only one third of CML patients were treated with imatinib, and the majority of the treated were not monitored in time. Clinicians should pay attention to resistance and intolerance to imatinib treatment in accelerated phase or blast crisis patients.

Benzamides ; therapeutic use ; China ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; Piperazines ; therapeutic use ; Pyrimidines ; therapeutic use