Adolescent ; Case-Control Studies ; Child ; Child, Preschool ; Heart Defects, Congenital ; blood ; immunology ; Humans ; Infant ; Lymphocyte Subsets ; Th1 Cells ; immunology ; Th2 Cells ; immunology

Objective To investigate the chemical constituents of Peperomia blanda. Methods Guided by HPLC detection, integrated methods including vacuum liquid chromatography (VLC), ODS and semi-preparative RP-HPLC were used for separation. The structure was determined by spectral analyses including ESI-MS, 1D NMR, 2D NMR and ECD spectra. Results Four compounds were isolated and identified as (7S,7’S,8R,8 ’R)-7-(5-methoxy-3,4-methylenedioxyphenyl)-7-(4-hydroxy-3,5-dimethoxy-phenyl)-8, 8-dihydroxymethyltetrahydrofuran (1), 7, 8-trans-8,8-trans-7,8-cis-7, 7-(4-hydroxy-3,5-dimethoxy phenyl)-8,8-di-acetoxymethyltetrahydrofuran (2), (+)-(7S, 7S, 8R, 8R)-4, 4-dihydroxy-3, 3, 5, 5-tetramethoxy-7, 9, 7, 9-diepoxylignane (3), and (6R, 7E, 9R)-9-hydroxy-4, 7-megastigmadien-3-one (4). Conclusion Compound 1 is a new tetrahydrofuran lignan, compounds 2-4 were isolated from P. blanda for the first time.

OBJECTIVERefractory temporal lobe epilepsy (TCE) shows a unique type of hippocampal damage, referred to as hippocampal sclerosis. The mechanisms underlying drug-refractoriness in TCE are poorly understood, which may be connected with pharmacoresistance to antiepileptic drugs (AEDs). Some studies show that expression of the multidrug resistance gene (mdr1a and mdr1b) and p-glycoprotein encoded by mdr1a and mdr1b are high in the brain, especially in the hippocampus, and the expression may lead to reduction of AEDs concentration in the brain. But most of these studies focused on acute epileptic activity shortly after status epilepticus (SE), spontaneous seizures are seldom studied. The authors used a rat model of kainic acid induced spontaneous seizures to investigate expression of mdr1a and mdr1b mRNA, and explore whether topiramate (TPM) affects expression of mdr1a and mdr1b in the hippocampus.

METHODSSeizures were induced by intraperitoneal injection of 10 mg/kg kainic acid at postnatal day 28. Control rats were injected with sodium chloride. All rats were divided into 4 groups 1 week after spontaneous seizures developed: status epilepticus complicated with spontaneous seizures (SE, n = 8) group, status epilepticus complicated with spontaneous seizures treated with TPM (SE + TPM, n = 9) group, spontaneous seizures without status epilepticus (N-SE, n = 7) group, spontaneous seizures without status epilepticus treated with TPM (N-SE + TPM, n = 8) group, control (n = 7) group and control treated with TPM (control + TPM, n = 7) group. The treated rats were given therapeutic dose of TPM (25 mg/kg). All the rats were killed on the 42nd day of administration. The mdr1a and mdr1b mRNAs in the hippocampus were measured by RT-PCR.

RESULTSExpression of mdr1a and mdr1b mRNA in the hippocampus increased significantly in the SE + TPM group, SE group and N-SE + TPM group compared with control group (P < 0.001 or < 0.05). The mRNA in SE + TPM group increased significantly compared with the SE group, too (P < 0.01). The mdr1a and mdr1b mRNA expression in the hippocampus in control + TPM and N-SE groups did not change.

CONCLUSIONFrequent seizures, especially status epilepticus resulted in overexpression of mdr1a and mdr1b mRNAs in the hippocampus. The drug-refractoriness mechanism in TCE may be related to overexpression of mdr1a and mdr1b mRNAs. TPM could enhance the expression of mdr1a and mdr1b mRNAs in the hippocampus. Seizure activity and TPM are likely to be the main determinant in enhancing mdr1a and mdr1b mRNA expression in epilepsy.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Animals ; Anticonvulsants ; pharmacology ; Fructose ; analogs & derivatives ; pharmacology ; Hippocampus ; metabolism ; Kainic Acid ; RNA, Messenger ; metabolism ; Rats ; Seizures ; chemically induced ; drug therapy ; metabolism ; Status Epilepticus ; drug therapy ; metabolism

3-Hydroxybutyric Acid ; blood ; Animals ; Dietary Carbohydrates ; administration & dosage ; Dietary Fats ; administration & dosage ; Dietary Proteins ; administration & dosage ; Disease Models, Animal ; Epilepsy ; diet therapy ; metabolism ; pathology ; Hippocampus ; metabolism ; Kainic Acid ; Ketone Bodies ; metabolism ; Male ; Mossy Fibers, Hippocampal ; pathology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, Kainic Acid ; analysis ; genetics

OBJECTIVEKetogenic diet (KD) is a high fat, low protein, low carbohydrate diet. Its antiepileptic effect is certain but the underlying mechanism is unknown. The aim of the study was to reveal the possible mechanism from the view points of synaptic reorganization and GluR(5) expression in hippocampus.

METHODSEpilepsy was induced in Sprague-Dawley rats by kainic acid at postnatal day 28, all control animals were fed with normal rodent chow, whereas experimental rats were fed with ketogenic feed for 8 weeks. Spontaneous recurrent seizures were recorded. Mossy fiber sprouting and neuron damage in hippocampus were investigated by Timm staining and Nissl staining. Western blot and RT-PCR methods were applied to detect the expression of GluR(5) and GluR(5) mRNA in hippocampus.

RESULTSKD-fed rats (1.40 +/- 1.03) had significantly fewer spontaneous recurrent seizures than control diet-fed rats (7.36 +/- 3.75). The mean A of mossy fiber sprouting in the inner molecular layer of dentate gyrus was markedly higher in KA induced animals than that in saline control animals but it was similar in different diet fed groups. No significant differences were found in the mean A of Timm staining in CA(3) area and Nissl staining of neuron in hilus, CA(3) and CA(1) area. After KA kindling, KD-fed animals [(189.38 +/- 40.03)/mg pro] had significantly higher GluR(5) expression in hippocampus than control diet-fed animals [(128.79 +/- 46.51)/mg pro] although their GluR(5) mRNA was the same.

CONCLUSIONMossy fiber sprouting may be responsible for epileptogenesis in KA induced model and KD can suppress seizures in these animals. KD may upregulate young rat GluR(5) in inhibitory interneurons of CA(1) thus lead to an increased inhibition to prevent the propagation of seizure.

Animals ; Blotting, Western ; CA1 Region, Hippocampal ; metabolism ; pathology ; CA3 Region, Hippocampal ; metabolism ; pathology ; Chromosome Pairing ; drug effects ; Dentate Gyrus ; metabolism ; pathology ; Diet, Ketogenic ; methods ; Disease Models, Animal ; Epilepsy ; chemically induced ; diet therapy ; genetics ; metabolism ; pathology ; Excitatory Amino Acid Agonists ; Hippocampus ; drug effects ; metabolism ; pathology ; Kainic Acid ; Male ; Mossy Fibers, Hippocampal ; metabolism ; pathology ; Pyramidal Cells ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Rats ; Receptors, Kainic Acid ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective Ketogenic diet(KD) is a high fat, low protein, low carbohydrate diet. Its antiepileptic effect is certain but the underlying mechanism is unknown. The aim of the study is to reveal the possible antiepileptic mechanism of KD treatment from the view points of synaptic reorganization and GluR5,GluR6 mRNA in hippocampus. Methods Sprague-Dawley rats were induced by kainic acid (KA) at postnatal day 28 as experimental group, control animals were injected the same volume of saline. Animals from each group were divided into two parts, each of which was fed normal rodent chow or ketogenic diet for 8 weeks. Spontaneous recurrent seizures were recorded. Spatial learning and memory ability was evaluated by Morris water maze. Mossy fiber sprouting and neuron damage in hippocampus were investigated by Timm staining and Nissl staining. RT-PCR method was applied to detect the expression of GluR5, GluR6 mRNA in hippocampus. Results KD-fed rats had significantly fewer spontaneous recurrent seizures(1.40±1.03) than control diet-fed rats(7.36±3.75). In water maze testing, all groups demonstrated improvement in water maze performance with significantly decreased escape latencies during the testing days(F=33.93 , P<0.001),but no significant differences were found in the time to platform among the four groups(F=1.24 , P=0.32). The mean A of mossy fiber sprouting in the inner molecular layer of dentate gyrus was markedly higher in KA-induced animals than that in saline control animals but it was similar in different diet-fed groups. No differences were found in the mean A of Timm staining in CA3 area and Nissl staining of neurons in hilus, CA3 and CA1 areas. In KA+KD group rats, GluR6 mRNA was statistically higher (48.16±11.53) than that in KA+ND group rats(30.57±15.57, t=2.40, P<0.05), but no significant difference of GluR5mRNA was found in these two groups (t'=-0.09, P>0.05). Conclusion KD had prominent antiepileptic effect on KA-induced rats and it had no significant impairment on spatial learning and memory for the developing brain. KD can not prevent the MFS in young KA-induced rats, but it may play its antiepileptic role by keeping the high level expression of GluR6 mRNA in the hippocampus, due to its specific neuroprotective action, to inhibit the excitatory transmission at the MF pathway.

OBJECTIVECleidocranial dysplasia (CCD) is a dominantly inherited skeletal dysplasia caused by mutations in the osteoblast-specific transcription factor-encoding gene, core binding factor α1 (CBFA1). Over 90 mutations in CBFA1 gene have been published to date in 500 independent cases of CCD, including missense mutations, deletions, insertions, frameshift, and splice mutations. However, mutational screening of the CBFA1 gene is still far from saturation, and more novel mutations will be identified to enrich the insights into the molecular basis for the pathogenesis of CCD. The aim of this study was to explore the clinical and image features and detect the mutations of CBFA1 gene in two CCD families.

METHODIn this study, the clinical features were investigated in two CCD families, radiological and CT examinations regarding osseous malformation were carried out over the entire body of these patients with CCD. Blood (2 ml) was drawn from all affected individuals, unaffected family members and one hundred unrelated normal controls, Genomic DNA was extracted from whole blood with PureGene DNA extraction kit and PCR was performed with eight pairs of PCR primers for exons 0 to 7 of the CBFA1 gene. The mutations of CBFA1 gene were screened in these two CCD families.

RESULT(1) The clinical features of patients with CCD include delayed closure of fontanelles, frontal bossing, dysplasia of clavicles, late tooth eruption, and other skeletal anomalies. X-ray and CT examination showed the bulging calvarium, patent fontanelles, wide cranial sutures, multiple Wormian bones, dental dysplasia or aplasia of clavicles. (2) Two mutations were identified, one is novel missense mutation (c.1259C > T[p.T420I]) in CBFA1 gene exon 7, other (c.577C > T[p.R193X]) was reported in Chinese cases with CCD for the first time.

CONCLUSION(1) The clinical and image features of patients in two CCD families include delayed closure of fontanelles, frontal bossing, dysplasia of clavicles, late tooth eruption, and other skeletal anomalies. (2) The T420I and R193X mutations of CBFA1 were reported, expanding the spectrum of CBFA1 mutations causing CCD.

Child ; Child, Preschool ; Cleidocranial Dysplasia ; genetics ; pathology ; Core Binding Factor Alpha 1 Subunit ; genetics ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Male ; Mutation ; Pedigree ; Phenotype

OBJECTIVETo study the chemical constituents from the whole plant of Dracocephalum moldavica.

METHODThe compounds were isolated by using column chromatography with RA polystyrene resin, polyamide and silica gel as packing materials, and the structures of the compounds were identified by means of spectral data.

RESULTeight compounds were identified as apigenin(I), luteolin(II), kaempferol(III), isorhamnetin(IV), tilianin(V), agastachoside(VI), acacetin-7-O-(6-O-Malonyl-beta-D-glucopyranoside) (VII) and syringaresinol(VIII).

CONCLUSIONCompounds I, II and III were isolated from genus Dracocephalum for the first time and compounds IV, VII and VIII were isolated from Dracocephalum moldavuca for the first time.

Apigenin ; chemistry ; isolation & purification ; Chromatography, Thin Layer ; Furans ; chemistry ; isolation & purification ; Lamiaceae ; chemistry ; Lignans ; chemistry ; isolation & purification ; Luteolin ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry

OBJECTIVETo study the chemical constituents from the bark of Morus macroura.

METHODThe compounds were isolated with silica gel column chromatography and their structures were elucidated on the basis of chemical evidences and spectral analysis (IR, EI-MS, 1H-NMR, 13C-NMR).

RESULTFive compounds were identified as alpha-amyrin acetate (1), gult-5-en-3 beta-yl acetate (2), 3 beta-hydroxylup-20 (29)-en-28-oic acid (3), 3 beta-hydroxylup-12-en-28-oic acid (4), butyrospermol acetate (5).

Morus ; chemistry ; Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Organic Chemicals ; Plant Bark ; chemistry ; Plant Extracts ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry

OBJECTIVE To study the changes of symptoms and signs of laryngopharyngeal reflux diseases(LPRD) after treatment with esomeprazole.METHODS The suspected LPRD patients were treated with esomeprazole for 8 weeks.Reflux finding score(RFS) and reflux symptoms index(RSI) score were evaluated before and after treatment.RESULTS In RSI,84％ of the patients had the symptom of hoarseness or a problem with voice,87％ had clearing of throat,71％had excess throat mucus,58％ had difficulty swallowing food,52％ had coughing after eat or after lying down,68％ had breathing difficulties or chocking episodes,79％ had troublesome or annoying cough,92％ had lump in your throat or sensation of something sticking in throat,and 32％ had heartburn,chest pain,indigestion or stomach acid coming up.In the RFS,45％ of the patients had pseudosulcus,57％ had ventricular obliteration,94％ had erythema/hyperemia,85％ had vocal cord edema,82％ had diffuse laryngeal edema,83％ had posterior commissure hypertrophy,10％ had granuloma/granulation,and 58％ had thick endolaryngeal mucus.The RSI and RFS score had statistical difference before and after treatment(P＜0.001).CONCLUSION The main symptoms of LPRD were lump in throat or sensation of something sticking in throat,clearing of throat and hoarseness or a problem with voice.The main signs of LPRD were erythema/hyperemia,vocal cord edema and posterior commissure hypertrophy.The PPI has marked improved in symptoms and signs of LPRD.