Aim A method of coculture of brain capillary endothelial cells (BCECs) and astrocytes of rats was used to evaluate nanoparticle' s blood-brain barrier (BBB) transcytosis and toxicity at the endothelial tight junction. Methods A lipophilic fluorescent probe, 6-coumarin, was incorporated in poly(ethyleneglycol)-poly (lactide) nanoparticle using double emulsion/solvent evaporation method. BCECs and astrocytes were firstly isolated from brain of newborn rats and characterized by their morphology and immunocytochemistry staining, separately. Subsequently, a coculture model with BCECs on the top of micro-porous membrane of cell culture insert and astrocytes on the bottom side was established. The permeability of 14C-labeled sucrose and nanoparticle were determined, separately. Results The meanweight-based diameter of 6-coumarin loaded nanoparticles was ( 102.4 ± 6.8) nm, with zeta potential of ( - 16.81 ± 1.05) mV. BCECs were positive for factor Ⅷ staining and glial fibrillary acidic protein was tight junction between BCECs in the coculture model could be visualized by both scanning electron microscopy and transmission electron microscopy. The unchanged paracellular transport of sucrose proved vivo situation for examination of the permeability of nanoparticle and toxicity evaluation.

Objective To explore the release and percutaneous penetration of cyclovirobuxine D patches at different concentrations in vitro. Methods The release curves of cyclovirobuxine D patch in vitro were fitted by ritger-peppas mathematical model, and the patch release mechanism was discussed according to the fitting parameters. At the same time, compared the percutaneous permeability characteristics of 0.25,0.5,1.0,2.0 mg.( cm2 )-1 of cyclovirobuxine D patch by using a modified Franz diffusion cell, with isolated rat skin serving as transdermal barrier. Results Ritger-Peppas model fitting equation for cyclovirobuxine D patch [1.00 mg.(cm2)-1]was: Mt/M=0.964 6 t1.621 6.And the percutaneous penetration curve was best fitted to Higuchi kinetics equation.The drug release rate from the patch matrix was greater than the rate of penetration through the skin, indicating the patch at the time through rat′s skin was a passive diffusion process, and transdermal process was rate-limited by skin. Conclusion Kinetics equation fitting is an effective method for analyzing drug release and permeation behavior of cyclovirobuxine D patch in vitro.

Purpose To optimize the preparation of NC-1900 loaded MePEG-PLA nanoparticles (NPs). Methods MePEG-PLA copolymers of different molecular weight synthesized by solvent poly-merization method were used to prepare NC-1900 loaded MePEG-PLA NPs by double emulsion/solvent evaporation method.Orthogonal experimental design and multiple regressions were used to optimize the preparation method with nanoparticle size and NC-1900 encapsulation efficiency (EE) as res-ponse variables.NPs were characterized by particle size and Zeta potential detector and transmission electron microscope.The leakage of NC-1900 from NPs was evaluated by high-performance liquid chromatography (HPLC) detection. Results MePEG3000-PLA44800 NPs prepared according to the optimized conditions had a mean diameter of (77 ± 11) nm and EE of (21.4 ± 0.1) %.Only 5% and 15% of NC-1900 were leaked in pH 7.4 PBS and blank plasma at the end of 48 h,respectively. Conc-lusions The optimized MePEG3000-PLA44800 NPs is a favorable carrier for NC-1900.

Brain Diseases ; diagnosis ; Child, Preschool ; Epilepsy ; diagnosis ; Female ; Fever ; Humans ; Infant ; Male ; Skin ; pathology ; Tomography, X-Ray Computed ; Tuberous Sclerosis ; diagnosis

Objective To assess the efficacy and safety of two different protocols of ketogenic diet (KD)-eating on demand or eating at regular intervals for refractory epilepsy in children.Methods Sixty children with refractory epilepsy were randomly divided into eating on demand group (n =30) and eating at regular intervals group (n =30) by random number table method.After taking the whole amount of KD,the capillary blood ketone and glucose level and urine ketone were monitored every 6 hours in 72 continuous hours.Seizure frequency and onset time were recorded.Antiepileptic efficacy and diet tolerability of the two groups were evaluated on 4 weeks,12 weeks,24 weeks and 48 weeks after initiating the diet.Adverse effects were monitored.Results After treatment of 4 weeks,the complete seizure remission rates of eating on demand group and eating at regular intervals group were 33.3％ (10/30) and 30.0％ (9/30) respectively,which suggested a comparable efficacy for two groups (P ＞ 0.05).The day when KD started to work was averaged (6.18 ± 2.42) d and (8.63 ± 2.63) d respectively.The group of eating on demand showed a faster onset of action (P ＜0.05).After treatment of 12 weeks,24 weeks and 48 weeks,complete seizure remission rates of eating on demand group were 30.0％ (9/30),34.8％ (8/23) and 36.8％ (7/19) respectively;the eating at regular intervals group were 33.3％ (10/30),30.4％ (7/23) and 44.4％ (8/18) respectively.The two groups had no significant difference (P ＞ 0.05).One year later,the treatment retention rates of the two groups were 63.3 ％ (19/30) and 60.0％ (18/30) respectively.There was no significant difference (P ＞ 0.05).The adverse effects mainly including transient gastrointestinal symptoms and metabolic disturbances were mostly tolerable and curable.Conclusion The two different protocols of KD-eating on demand and eating at regular intervals are both effective and well-tolerated for refractory epilepsy in children.While protocol of eating on demand is more easier to achieve ketotic state and the effect is more quickly,so it can be more easily received by children.Therefore in clinical practice,we can choose flexible eating time according to children's eating habits,which can improve the therapeutic compliance.

Objective To analyse the value of video-EEG (VEEG) in the infantile spasms (IS)children treated with adrenocorticotrophic hormone (ACTH). Methods The clinical data of 65 children with IS in our hospital from Jan 2005 to Dec 2009 were collected and the characteristics of VEEG were analyzed before and after treated with ACTH. Results All the 65 cases appeared hypsarrhythmia in the interphase before treated with ACTH. Cluster spasms and the characteristics of VEEG were recorded. After treated with ACTH,40 of 65 cases (61.5%) with epileptic seizure were controlled,in whom 27 cases (67.5% ,27/40)with hypsarrhythmia disappeared or improved obviously. The other 25 of 65 cases (38.5%) with epileptic seizure were uncontrolled,in whom only 5 cases (20. 0% ,5/25) with hypsarrhythmia disappeared. The rate of hypsarrhythmia disappeared or improved was significantly different between the epileptic seizure controlled and uncontrolled children (x2 = 13. 888, P ＜ 0. 000). Conclusion VEEG is not only important to make a definite diagnosis for IS,but also can provide an evidence in evaluating the effect of ACTH in IS children.

Objective To explore the common pathogen of infantile convulsions associated with mild gastroenteritis, and to study the differences between the seizures caused by the two kinds of virus.Methods RT-PCR was used to detect Rotavirus (RV) and Norwalkvirus (NoV) in stool and cerebrospinal fluid of 30 cases with infantile convulsions associated with mild gastroenteritis. The differences between the frequency of seizures caused by two kinds of virus were analyzed by statistical methods (two-sample t-test).Results 17/30 (56. 7%) were RV-positive in stool and 3/17 (17. 7%) in cerebrospinal fluid; 6/30 (25.0%) were NoV-positive in stool and 1/6 (16. 7%) in cerebrospinal fluid. The seizure frequency with NoV infection was (4. 33 ± 1.75) times, and RV infection patients was (2. 53 ± 1.12) times (P ＜ 0. 01).The seizure frequency of CSF virus-positive children was (4. 75 ± 1.71) times compared to (2. 63 ± 1.21)times in virus-negative children (P ＜ 0.01). Conclusion The common pathogens causing infantile convulsions associated with mild gastroenteritis were RV and NoV. The degree of NoV infection affecting the central nervous system may be greater than RV. The presence of the virus in cerebrospinal fluid may lead to higher incidence of seizures,but their exact roles related to the occurrence of seizures remain to be further studied.

[Objective] To learn about the genetic diversity,we studied the five X-chromosomal STR (X-STR) loci in Guangdong Han Nationality Groups.[Methods] The five Loci (DXS6803,DXS981,DXS6809,DXS6789,and DXS7132) were amplified in a pentaplex PCR reaction.PCR products were analyzed using capillary electrophoresis and ABI prism 3100 Genetic Analyzer,with GeneMapper ID 3.1 Analysis Software.[Results] A total of 363 individuals (181 unrelated male and 182 unrelated female) from Guangdong Han population were tested,54 alleles were observed for these loci.Polymorphism information content is 0.6935 ～ 0.8177.Power of discrimination in females was 0.8976 ～ 0.9562.Mean exclusion chance for X-STR in standard trios with daughters was 0.7805 ～ 0.8467.[Conclusion] The five loci in the multiplex system provide high polymorphism information for forensic identification and paternity testing,particularly for difficult paternity deficiency cases.

Objective We aimed to investigate whether magnetic stent has preventive effect on in-stent restenosis by observing expressions of matrix metalioproteinase (MMP)2,MMP9,tissue inhibitor of matrix metalloproteinase (TIMP)1 and TIMP2 after balloon angioplasty,bare and magnetic stent implantation in rabbits.Methods Rabbits underwent balloon angioplasty,bare and magnetic stent implantation in the left iliac arteries.The changes of MMPs and TIMPs were examined at various time points in the injured arteries using the methods of zymography,Western blot analysis,reverse transcription-polymerase chain reaction (RT-PCR) and morphometric analysis.Results Balloon angioplasty group (BA) and magnetic stent group (MS) showed lower intrinsic gelatinolytic activity and higher expression of TIMPs with less intimae hyperplasia;Whereas bare stent (BS) group exhibited higher intrinsic gelatinolytic activity and lower expression of TIMPs with significant intimae hyperplasia.Conclusion Magnetic stent probably has preventive effect on in-stent restenosis by changing intrinsic matrix metalloproteinases activity and expression of TIMPs.

Objective To investigate the preventive effect of magnetic stent on coronary restenosis after percutaneous arterial stenting.Methods Twenty rabbits were divided randomly into 2 groups.Bare stent(BS group,n=10)or magnetic stent(MS group,n=10)wasimplanted in the left iliac artery of the rabbits of the 2 groups,respectively.Aspirin (25mg,qd )was administered orally to the rabbitsof both groups from 3 days before stenting until the rabbits were executed.Unfractionated heparin (2500u,qd) was delivered subcuta-neously after stenting for 7days.Five rabbits of each group were randomly selected to be executed at 7 or 30 days.Stmctural changesin the iniured arteries were studied by optical microscopey,transmissive electronic microscopey and immunohistochemistry.ResultsAt 7 days.more myofibroblasts were found migrating from adventitia to tunica media and intima in BS group than in MS group.insidethe media and intima,large amount of smooth muscle cells of synthetic type were observed.At 30 days after stenting,in magnetic group,most uascular smooth musele cells(SMCs)under the intima had transformed to contractile type and only little extracellular matrix(ECM)was observed around the SMCs;whereas,in BS group,the SMCs remained to be synthetic type and large amount of ECM wasobserved around the SMCs.which was composed mainly of proteoglycans and glycoproteins. Conclusions Magnetic stent caninhibit proliferation and migration of SMCs and reducing the production of ECM.and therefore,may prevent restenosis after coronarystenting.