Objective To investigate the effects of extracellular-signal regulated kinase (ERK) on hypoxic-ischemic brain damage of neonatal rats.Methods The hypoxic-ischemic brain damage model of neonatal Wistar rats was established as following:first the right common carotid artery of the rats was ligated;2 h after operation,the rats began to inhale 8%-oxygen oxygen-nitrogen gas mixture lasting for 2 h.Rats were randomly divided into three groups:sham-group,hypoxic-ischemic group and ERK inhibitor PD98059 group (the rats were injected PD98059 2 mg/kg 10 min before the ligation).Six hours after the models were done,hippocampi and cortex of the ligation side of rats in the three groups were collected,and the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) were measured.Apoptosis of neuron was assessed by TUNEL staining.The expression of ERK1,ERK 2,Bcl-2 and Bax were examined by Western blot.The differences among the groups were analyzed with ANOVA and q test.Results Compared with the sham-group,the MDA level [(342.9± 10.8) μmol/L vs (181.5± 17.0) μmol/L,q= 6.35,P＜0.01) and the apoptosis rate of neuron [(18.80±1.37)% vs (3.53±0.34)%,q=6.06,P＜0.01) of hypoxic-ischemic group was higher,and SOD level was lower [(34.8±4.3) U/ml vs (63.4±4.3) U/ml,q=4.99,P＜0.01].While the apoptosis rate of neuron [(15.53±0.64) %] and MDA level [(252.0± 17.1) μmol/L] of PD98059 group were lower than those of hypoxic-ischemic group(q=3.87 and 5.28,P＜0.01respectively),the SOD level [(51.5 ± 3.8) U/ml] was higher than that of hypoxic-ischemic group (q=4.17,P＜0.01).There were no differences of ERK1 and ERK2 expressions among the three groups.The phosphorylated ERK1 and ERK2 levels of hypoxic-ischemic group were higher than those of sham-group (q=3.82 and 4.08,P＜0.01) and PD98059 group (q=4.79 and 5.12,P＜0.01).The expression of Bcl-2 and Bax of hypoxic-ischemic group were higher than those of sham-group (q=3.55 and 3.42,P＜0.01).Compared with hypoxic-ischemic group,Bcl-2 expression (q=3.71,P＜0.01) of PD98059 group was higher,and Bax expression (q=5.86,P ＜ 0.01) was lower.Conclusions ERK is involved in hypoxic-ischemic brain damage of neonatal rats through regulating the expression of apoptosis protein.

Objective:To investigate the effects of Astragalus on the the apoptosis of cultured human peritoneal mesothelium induced by peritoneal dialysis solution(PDS). Methods:Using RPM1640 culture medium as control,PDS group(commercial PDS containing 1.5% glucose without Astragalus),Astragalus group 1(commercial PDS containing 1.5% glucose and 1% Astragalus) and Astragalus group 2(commercial PDS containing 1.5% glucose and 2% Astragalus) were tested to explore the apoptosis of cultured human peritoneal mesothelial cell in each group.The activity of caspase-3,the terminal deoxynucleotidyl transferase(TdT) mediated dUTP nick end labeling(TUNEL) and flow cytometry analysis were used in apoptosis analysis in this study. Results: The activity of caspase-3 in the control group was defined as 1,the relative activity of caspase-3 in PDS group、 Astragalus group 1 and Astragalus group 2 were 3.26?0.91,(1.87?)0.43 and 1.67?0.32 respectively.The activity of caspase-3 in PDS group was significantly higher than those in the Astragalus group 1 and Astragalus group 2(P

Objective:To investigate the impact of Puerarin on oxidative stress in patients undergoing continuous ambulatory peritoneal dialysis(CAPD).Methods: Thirty-eight patients CAPD were randomly divided into two groups,an experimental group(n=19) receiving peritoneal dialysis fluid containing Puerarin(50 mg/2L) in vivo for two weeks and a control group(n=19) receiving dialysis fluid without Puerarin.The concentrations of oxidative stress markers in the serum and effluent such as glutathione(GSH),total-superoxide dismutase(T-SOD),malondialdehyde(MDA),Kt/V and creatinine clearance rate of the two groups(Ccr) were measured and compared before and after the treatment.Results: After the treatment,the concentrations of GSH and T-SOD in the serum and effluent were significantly higher(P

Objective: To investigate the effect of Astragalus on peritoneal macrophages function in stable continuous ambulatory peritoneal dialysis(CAPD) patients. Methods: 28 stable CAPD patients were included in this study.All the patients were treated with peritoneal dialysis fluid containing Astragalus(20 ml/2 L) in vivo for one week.Peritoneal macrophages function such as phagocytosis ratio、bactericidal ratio、excretion of cytokines(TNF-? and NO) and mitochondrial dehydrogenases activity(MTT assay) were examined ex vivo just before and after the treatment. In vitro,peritoneal macrophages isolated from effluent were incubated with RPMI 1640、CDF or CDF containing various concentrations of Astragalus. Then peritoneal macrophages function mentioned above were examined again and compared. Results:No significant difference was found in TNF-? level in effluent between before and after the treatment (100?63)ng/L vs (116?60)ng/L, (P=0.192). While comparing with pre-treatment, peritoneal macrophages function was improved significantly after the addition of Astragalus into dialysate (phagocytosis ratio: (34.8?12.7)% vs (43.4?9.3)%,P0.05).The results in these two groups were significantly lower than those in Astragalus groups(P0.05). While significant decreasing in NO level and mitochondrial dehydrogenases activity were observed in 8% Astragalus groups as compared with those in 2% Astragalus group(P

Objective: To investigate the effect of Astragalus mongholicus injection(AM) on the secretion and mRNA expression of transforming growth factor((TGF-?1)) and basic fibroblast growth factor(bFGF) in cultured human peritoneal mesothelial cells((HPMC).) Methods: HPMC got from patients underwent surgical operation were cultured in vitro.At first,cells from the third passage were incubated with RPMI1640 culture medium containing 0.1% FBS for 24 hours.Then,they were divided into control group,PDS group,AM group 1,AM group 2 and AM group 3 for testing.Each group was supplemented with equal volum of RPMI1640 culture medium containing 20% FBS.After 24 hour incubating,mitochondrial dehydrogenases activity(MTT assay),the levels of TGF-?1 and bFGF in supernatant of the cell culture and the mRNA expression of TGF-?1 and bFGF were detected.Results: Significant decrease of mitochondrial dehydrogenase activities were observed in PDS group as compared with those in control group and AM groups(P0.05).The levels of TGF-?1 and bFGF in supernatant of the cell culture were significantly lower in control group than those in PDS group.Marked lower levels of TGF-?1 and bFGF were found in AM group 1,AM group 2 and AM group 3 as compared with those in PDS group(P0.05).The mRNA expressions of TGF-?1 and bFGF in PDS group increased significantly as compared with those in control group.And significant mRNA expressions of TGF-?1 and bFGF were found in PDS group when compared with those in AM groups(P

The medical law science interdisciplinary talents play an crucial role in solving medical service dispute and alleviating contradictory. Based on the questionnaire survey and the interview to the court and law offices as well as the analysis of the society's demand for the knowledge structure and ability of medicine law professionals, we proposed the construction of the training mode for medicine law professionals with the ability to adapt to the social demand.

ObjectiveToexploretheeffectof ceremideonprocess of peritoneal mesothelial cells(PMCs) apoptosis induced by peritoneal dialysis solution(PDS).Methods PMCs were cultured with normal DMEM,1.5％ PDS and 4.25％ PDS.4.25％ mannitol was used as high osmotic pressure control.Ceremide were detected by LC-MS-MS.Flow cytometry was used in apoptosis analysis.Bax,p53 and bcl-2 protein expressions were detected by Western blotting.Results (1) PDS caused the increase of intracellular ceremide in PMCs,and normal and high osmotic pressure controls had no such effect.As the acidic sphigomyelinase inhibitor,desipramine significantly inhibited the production of ceramide induced by 4.25％ PDS [(56.08±12.24) μg/L vs (91.25:t:15.89) μg/L,P＜0.01]. (2) Compared with 1.5％ PDS,4.25％ PDS stimulated PMCs apoptosis (26.65％±6.21％ vs 4.04％±1.86％,P＜0.01),up-regulated bax and p53 proteins expression (P＜0.01),and down-regulated bcl-2 protein exprssion(P＜0.05).Desipramine obviously inhibited the apoptosis induced by 4.25％ PDS,decreased bax and p53 proteins expression,increased bcl-2 protein expression(P＜0.05).Exogenous C2-ceremide reversed the effect of desipramine(P＜0.05).Conclusion The increase of intracellular ceremide may play an important role in the PMCs apoptosis induced by high glucose PDS.

Objective Calcium-sensing receptor(CaSR)belongs to the family C of G-protein coupled receptors.This study was carried out to observe the influence and mechanism of CaSR on anoxia/reoxygenation(A/R)-induced cardiomyocytes apoptosis.Methods The model of A/R injury was established through anoxia for 2 hours and reoxygenation for 24 hours in cultured cardiomyocytes of neonatal rats.Cardiomyocytes were randomly divided into three groups:control group,A/R group and GdCl3 group(300μmol/L GdCl3 was added to the culture medium at the beginning of reoxygenation).Apoptosis of cardiomyocytes was assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL).The expression of CaSR,cysteine-requiring aspartate protease(caspase)-3,9 and cytochrom c (Cytc)were analyzed by Western blot.Results The TUNEL showed that cardiomyocytes apoptosis was 17%±3% in A/R group,and was higher than that in the control group.At the same time,expression of CaSR in A/R group was markedly increased in response to control group.Compared with A/R group,GdCl3,a specific activator of CaSR,further enhanced cardiomyocytes apoptosis to (28±4)% and decreased the ability of cardiomyocytes to (51.2±6.8)%,along with an increment in CaSR,caspase-3,caspase-9 and Cytc expressions.Conclusion CaSR is involved in anoxia/reoxygenation-induced apoptosis of neonatal rat cardiomyocytes through Cytc-caspase-3 pathway.

ObjectiveTo observe the influence of calcium - sensing receptor (CaSR) on Fas/Fas ligand (Fas L) pathway during anoxia/reoxygenation (A/R)- induced cardiomyocytes apoptosis in neonatal rat. MethodsSingle cells were dissociated from minced hearts of 2 - day - old Wistar rats with a 0. 25％ solution of crude trypsin and then cultured as monolayers at a density of 5 x 104cells/cm2 in DMEM medium equilibrated with humidified air containing 5％ CO2 at 37C. Three days after the cells were seeded, the cultured cardiomyocytes were randomly divided into three groups. ( 1 ) control group: cardiomyocytes were continuously cultured for 26 hours in DMEM medium.(2) A/R group: cardiomyocytes underwent anoxia for 2 hours and reoxygenation for 24 hours.(3) GdCl3 group: 300μmol/L GdCl3 was added to the culture medium at the beginning of reoxygenation. Apoptosis of cardiomyocytes was assessed by flow cytometer and morphological alterations were observed with transmission electron microscope.The expression of CaSR, Fas and Fas L were analyzed by Western blot.Results The result of flow cytometer showed that cardiomyocytes apoptosis was 12. 18％ ± 1.54％ in A/R group,and was higher than that in the control group (P ＜0. 01 ). At the same time, mitochondrial cristae dissolution and disappearance could be detected. Compared with A/R group, GdCl3, a specific activator of CaSR, further enhanced cardiomyocytes apoptosis to 20. 25％ + 2. 87％ ( P ＜ 0. 01 ), along with an increment in CaSR, Fas and Fas L expressions ( P ＜ 0. 05 ). ConclusionCaSR is closely involved in cardiomyocytes apoptosia during anoxia/reoxygenation. CaSR could induce apoptosis of neonatal rat cardiomyocytes through Fas/Fas L receptor pathway.

Objective To explore the clinical manifestations, electroencephalographic characteristics and therapeutic effect of drugs in children with Jeavons syndrome. Methods The clinical and electroencephalographic characteristics and thera-peutic effect of drugs were analyzed in 4 children with Jeavons syndrome. Results Among the four children there were 3 female and 1 male. The age at the onset of the disease was from 1 to 6 years. The typical clinical manifestations of this disease were brief, fast and repeated eyelid myoclonia (EM) with or without absence seizure. The typical electroencephalography (EEG) in two patients showed 3-6 Hz generalized spike and waves and polyspikes burst, and the eye closure and intermittent photic stimu-lation helped to induce discharges and clinical events. The typictal EEG in the other two patients showed 3.0-3.5 Hz generalizedδslow wave rhythm burst. The drugs of choice for treatment was sodium valproate monotherapy in two cases, levetiracetam in one case, sodium valproate combined with levetiracetam in one case. During the follow-up, seizures were controlled in one case, decreased in frequency in two cases and were still frequent in one case. Conclusions Jeavons syndrome is one of the idiopathic and generalized epileptic syndromes and characterized by EM with or without absence seizure. Video EEG monitoring plays an important role in the diagnosis of this disease. Sodium valproate and levetiracetam were effective for this disease.