Clinical and genetic characteristics of epilepsy of infancy with migrating focal seizures in Chinese children.

OBJECTIVE: Epilepsy of infancy with migrating focal seizures (EIMFS) is a rare and severe developmental epileptic encephalopathy. The aim of this study was to improve our understanding of EIMFS by using phenotype-genotype correlation. METHODS: We recruited, performed clinical genetic testing, and summarized the clinical features and genetic characteristics in five patients with EIMFS in China. RESULTS: The five recruited patients included 2 males and 3 females. The median age of seizure onset was 2 months (range, day 3 to 3 months). All patients exhibited the characteristics of clinically migrating focal motor (tonic or clonic) seizures. Typical migrating ictal electrical patterns were found in 1 patient; the remaining four patients presented with overlapping seizures with different areas of ictal onset in differing hemispheres. All the patients had the associated variants, including KCNT1, SCN1A, SCN2A, TBC1D24 and ALG1. All patients received two or more antiseizure medications, and 1 patient became seizure-free, 1 reported >75 % seizure reduction, 2 reported >50 % seizure reduction, and 1 patient showed no improvement. Varying degrees of psychomotor developmental delays were observed in all patients. CONCLUSIONS: The course of EIMFS could be related to the type of gene variant present, and different genes may have specific clinical features. Larger cohorts are required to elucidate such potential phenotype-genotype correlations.

[DuzhongButiansu Capsules improve adenine-induced reproductive dysfunction in male rats].

OBJECTIVE: To study the effect of DuzhongButiansu Capsules (DBC) on adenine-induced reproductive dysfunction (RD) in male rats. METHODS: Eighty male SD rats were randomly divided into six groups, blank control (n = 8), solvent control (n = 8), RD model control (n = 16), Shengjing Capsules (SJC) (n = 16), low-dose DBC (n = 16) and high-dose DBC (n = 16). The RD model was made by intragastric administration of adenine at 200 mg/kg/d for 5 successive weeks in the latter four groups of animals, and in the meantime the rats in the latter three groups were treated intragastrically with SJC at 0.560 mg/kg/d and DBC at 0.242 and 0.968 mg/kg/d, respectively. At the end of the fourth week, all the rats were mated with female ones in a 1:1 ratio for 7 days. Then the male rats were killed and the right epididymides collected for detection of sperm concentration and motility, and the female ones sacrificed after fed for another 2 weeks and the numbers of pregnancies and fetal rats were recorded. The heart, liver, spleen, lung, kidney, thymus, testis, epididymis and seminal vesicle were harvested for obtainment of the visceral coefficients and semen parameters, observation of the histopathological changes in the testis, epididymis and kidneys by HE staining, measurement of the levels of serum T, E2, FSH and LH by ELISA, detection of the contents of serum glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), malondialdehyde (MDA) and creatinine (Scr), blood urea nitrogen (BUN), and determination of the expressions of Bax, Bcl-2, Caspase-3 and Caspase-9 proteins in the renal tissue by immunohistochemistry. RESULTS: No statistically significant difference was observed between the blank control and solvent control groups in any of the indexes obtained (P > 0.05).Compared with the blank controls, the rats in the RD model control group showed significantly decreased sperm concentration (ï¼»40.67 ± 7.37ï¼½vs ï¼»27.10 ± 2.72ï¼½ ×106/ml, P < 0.01), sperm motility (ï¼»54.75 ± 3.92ï¼½%vs ï¼»25.60 ± 4.83ï¼½%, P < 0.01) and pregnancy rate (85.7% vs 43.8%, P < 0.01). The rats in thelow- and high-dose DBCgroups exhibited remarkable increases in sperm concentration (ï¼»53.00 ± 4.55ï¼½% and ï¼»65.63 ± 12.47ï¼½% ×106/ml, P < 0.01) and sperm motility (ï¼»53.50 ± 8.83ï¼½% and ï¼»54.33 ± 7.92ï¼½ %, P < 0.01), and so did those in the high-dose DBC group in pregnancy rate (54.5%, P < 0.01).After medication, the animals showed markedly increased body weight and visceral coefficients of the testis, epididymis and seminal vesicle (P < 0.05 or P < 0.01), recovered morphology of the testis, epididymis and kidneys, reduced levels of Scr, BUN, FSH, LH and MDA in the serum (P < 0.05 or P < 0.01), increased contents of T, SOD and GSH-PX (P < 0.05 or P < 0.01), down-regulated expressions of Bax, Caspase-3 and Caspase-9 and up-regulated expression of Bcl-2 in the renal tissue (P < 0.05 or P < 0.01). CONCLUSIONS: DBC can improve adenine-induced reproductive dysfunction in male rats, which may be attributed to its effects of inhibiting the apoptosis of proteins, improving oxidative stress and elevating the levels of reproductive hormones.

Efficacy and safety of levetiracetam in the off-label treatment of neonatal seizures.

Background: Treatment of neonatal seizures includes etiotropic and anticonvulsant treatments. However, anticonvulsant use in neonates is off-label and requires ethical review.Objective: To investigate the efficacy and safety of levetiracetam for neonatal seizures and to establish a predictive model.Methods: We retrospectively analyzed 125 neonatal seizure cases (phenobarbital 66 cases, levetiracetam 59 cases). The efficacy, safety and tolerability of levetiracetam were evaluated by cox regression survival analysis and a regression tree prediction model for the 16-week time point.Results: There was no significant difference between phenobarbital and levetiracetam treatment group in short-term efficacy (p > 0.05). But the cumulative survival function suggested that levetiracetam treatment group was better than phenobarbital (p = 0.026) in long-term efficacy evaluation. Neurodevelopmental assessments at 16 weeks showed that levetiracetam had better effect on the neurodevelopmental level (Gesell scores in response) than phenobarbital (p = 0.011). The main adverse events with levetiracetam were irritability and anorexia. According to the regression tree prediction model, the top three factors influencing the therapeutic effect were pre-treatment seizure frequency, age of onset and etiological classification.Conclusion: Levetiracetam shows good efficacy, safety and tolerability for the long-term neonatal seizure treatment.

Asperpyrone F, a new dimeric naphtho-γ-pyrone from the edible fungus Pleurotus ostreatus.

A new dimeric naphtho-γ-pyrone, asperpyrone F (1), along with six known ones, asperpyrones B (2) and C (3), fonsecinones A (4) and B (5), aurasperones A (6) and E (7), have been isolated from the solid culture of the edible fungus Pleurotus ostreatus. The structures of 1-7 were determined mainly by NMR and MS experiments. The absolute configuration of compound 1 was assigned via the circular dichroism (CD) data analysis. Compounds 1-7 showed modest antioxidant and immunomodulatory activities. All compounds were isolated from the fungus P. ostreatus for the first time.

Demonstration and reference of DRGs case management in a medical center in Taiwan of China / 中华医院管理杂志

Since the first edition of Tw-DRGs was developed in Taiwan in 2002, the system introduction of the hospital management level has been successfully completed. This paper detailed the organizational structure of DRGs case management in local hospitals ( DRGs promotion group, front-end medical care, ICD end, and the medical expenses reporting). It also introduced the operation process of identifying, locking and adjusting process of the pre-hospital, in-hospital and post-hospital aspects of inpatients, and provides in the end useful experiences for the promotion of DRGs case management in China′s mainland hospitals.

Effects of gap junctions blockers on fast ripples and connexin in rat hippocampi after status epilepticus.

OBJECTIVE: To assess the dynamic changes in the average and peak spectral power of fast ripples (FRs) in the hippocampi after interventions with valproate sodium (VPA), carbenoxolone (CBX) and quinine (QUIN). METHOD: Adult rats were used to establish a lithium-pilocarpine (pilo) epileptic model, and were assigned to a lithium-pilocarpine (PILO), VPA + PILO, QUIN + PILO or CBX + PILO group. Intracranial electroencephalography was performed before and after status epilepticus (SE). The hippocampal connexin (CX) 43, CX32 and CX36 expressions were analyzed via western blotting. RESULTS: The time required for the disappearance of SE after chloral hydrate injection was lower in the intervention groups than in the PILO group (p < 0.05). Seizures induced CX43 expression, but had no significant effects on CX36 or CX32 expressions. Pretreatment with VPA, QUIN and CBX inhibited CX43, CX36 and CX32 expression after SE. The average spectral power of the FRs was significantly lower in the VPA + PILO and QUIN + PILO groups than in the PILO group at 10 min after SE, 10 min before chloral hydrate injection, and 10 min after chloral hydrate injection (p < 0.05). The average spectral power of FRs was lower in the CBX + PILO group than in the PILO group at 10 min after SE (p < 0.05). The average spectral power of FRs in the 3 intervention groups recovered to the baseline level at 10 min after chloral hydrate injection and persisted for 3 days after SE. The dynamic changes in the average and peak spectral power of FRs were similar. SIGNIFICANCE: After SE, CX may participate in pathological FR generation by establishing abnormal electrical synaptic transmission. Gap junction blockers can inhibit various CX expressions, and thus decrease FR energy and alleviate the degree of seizure. These findings could contribute to the development of new anti-epileptic drugs with novel mechanistic targets.

Dynamic Changes in Spectral and Spatial Signatures of High Frequency Oscillations in Rat Hippocampi during Epileptogenesis in Acute and Chronic Stages.

OBJECTIVE: To analyze spectral and spatial signatures of high frequency oscillations (HFOs), which include ripples and fast ripples (FRs, >200 Hz) by quantitatively assessing average and peak spectral power in a rat model of different stages of epileptogenesis. METHODS: The lithium-pilocarpine model of temporal lobe epilepsy was used. The acute phase of epilepsy was assessed by recording intracranial electroencephalography (EEG) activity for 1 day after status epilepticus (SE). The chronic phase of epilepsy, including spontaneous recurrent seizures (SRSs), was assessed by recording EEG activity for 28 days after SE. Average and peak spectral power of five frequency bands of EEG signals in CA1, CA3, and DG regions of the hippocampus were analyzed with wavelet and digital filter. RESULTS: FRs occurred in the hippocampus in the animal model. Significant dynamic changes in the spectral power of FRS were identified in CA1 and CA3. The average spectral power of ripples increased at 20 min before SE (p < 0.05), peaked at 10 min before diazepam injection. It decreased at 10 min after diazepam (p < 0.05) and returned to baseline after 1 h. The average spectral power of FRs increased at 30 min before SE (p < 0.05) and peaked at 10 min before diazepam. It decreased at 10 min after diazepam (p < 0.05) and returned to baseline at 2 h after injection. The dynamic changes were similar between average and peak spectral power of FRs. Average and peak spectral power of both ripples and FRs in the chronic phase showed a gradual downward trend compared with normal rats 14 days after SE. SIGNIFICANCE: The spectral power of HFOs may be utilized to distinguish between normal and pathologic HFOs. Ictal average and peak spectral power of FRs were two parameters for predicting acute epileptic seizures, which could be used as a new quantitative biomarker and early warning marker of seizure. Changes in interictal HFOs power in the hippocampus at the chronic stage may be not related to seizure occurrence.

Synthetic promoters consisting of defined cis-acting elements link multiple signaling pathways to probenazole-inducible system.

Probenazole (3-allyloxy-1,2-benzisothiazole-1,1-dioxide, PBZ), the active component of Oryzemate, could induce systemic acquired resistance (SAR) in plants through the induction of salicylic acid (SA) biosynthesis. As a widely used chemical inducer, PBZ is a good prospect for establishing a new chemical-inducible system. We first designed artificially synthetic promoters with tandem copies of a single type of cis-element (SARE, JERE, GCC, GST1, HSRE, and W-box) that could mediate the expression of the ß-glucuronidase (GUS) reporter gene in plants upon PBZ treatment. Then we combined different types of elements in order to improve inducibility in the PBZ-inducible system. On the other hand, we were surprised to find that the cis-elements, which are responsive to jasmonic acid (JA) and ethylene, also responded to PBZ, implying that SA, JA, and ethylene pathways also would play important roles in PBZ's action. Further analysis demonstrated that PBZ also induced early events of innate immunity via a signaling pathway in which Ca(2+) influx and mitogen-activated protein kinase (MAPK) activity were involved. We constructed synthesized artificial promoters to establish a PBZ chemical-inducible system, and preliminarily explored SA, JA, ethylene, calcium, and MAPK signaling pathways via PBZ-inducible system, which could provide an insight for in-depth study.

Post-harvest induced production of salvianolic acids and significant promotion of antioxidant properties in roots of Salvia miltiorrhiza (Danshen).

Danshen, the dried roots of Salvia miltiorrhiza, is an extremely valued Traditional Chinese Medicine. Previously, we have demonstrated that salvianolic acid B (SaB), the important bioactive ingredient in this herb, was a post-harvest product. Here, we further reported that all salvianolic acids (SAs) in the roots were post-harvest products of the drying process. In addition, the results of various radical scavenging activity assays, including lipid peroxidation (1), DPPH (2), hydroxyl (3) and superoxide (4), were significantly increased along with the accumulation of total salvianolic acids in the process. The contents of chemical targets and antioxidant activities both reached the highest value under thermal treatment at 130 °C for 80 min. In this dehydration period, contents of SaB, and sum of nine SAs increased from 0.01% to 5.51%, and 0.20% to 6.61%; and IC50 of antioxidant activity decreased from 4.85 to 2.69 (1); 7.75 to 0.43 (2); 2.57 to 1.13 (3) and 17.25 to 1.10 mg/mL. These results further supported the hypothesis that the newly harvested plant roots were still physiologically active and the secondary metabolites might be produced due to dehydration stress after harvest. Our findings supplied an important and useful theoretical basis for promoting the quality of Danshen and other medicinal plant materials.

[Investigations of fluorescence spectral correction for a two-component mixed solution with overlapping absorption spectra].

The inner filter effect (IFE) and the absorption depth from the fluorescence material will directly lead to an artifact for intensity and spectral shape of the fluorescence spectra, and they restrict the application of fluorescence analysis. A two-component mixed solution with overlapping absorption spectra was used to develop a new method based on physical absorption model to correct the influences of IFE and the absorption depth on fluorescence intensity. The spectral investigations of terthiophene/ quinquethiophene mixture solution show that a good correction result can be obtained by using the above correction method, and the error after correction is less than 5 %.

Regulation of leaf senescence and crop genetic improvement.

Leaf senescence can impact crop production by either changing photosynthesis duration, or by modifying the nutrient remobilization efficiency and harvest index. The doubling of the grain yield in major cereals in the last 50 years was primarily achieved through the extension of photosynthesis duration and the increase in crop biomass partitioning, two things that are intrinsically coupled with leaf senescence. In this review, we consider the functionality of a leaf as a function of leaf age, and divide a leaf's life into three phases: the functionality increasing phase at the early growth stage, the full functionality phase, and the senescence and functionality decreasing phase. A genetic framework is proposed to describe gene actions at various checkpoints to regulate leaf development and senescence. Four categories of genes contribute to crop production: those which regulate (I) the speed and transition of early leaf growth, (II) photosynthesis rate, (III) the onset and (IV) the progression of leaf senescence. Current advances in isolating and characterizing senescence regulatory genes are discussed in the leaf aging and crop production context. We argue that the breeding of crops with leaf senescence ideotypes should be an essential part of further crop genetic improvement.

Ectopic overexpression of AtHDG11 in tall fescue resulted in enhanced tolerance to drought and salt stress.

Tall fescue (Festuca arundinacea Schreb.) is a cool-season perennial grass, which has been conventionally grown in the temperate area. However, as a major type of cool-season turf grass, its growth has been extended to the sub-tropical climate or even to the transitional climate between the sub-tropical and the tropical, and, in some cases, to heavily salinized lands. The extended growth imposes a serious challenge to its tolerance to the abiotic stress, particularly to drought, salt and high temperature. Here, we report a successful introduction of Arabidopsis AtHDG11 into the tall fescue via Agrobacterium-mediated transformation. The ectopic overexpression of AtHDG11 under the control of CaMV 35S promoter with four enhancers resulted in significantly enhanced tolerance to drought and salt stress. No obvious adverse effects on growth and development were observed in the transgenic plants. The enhanced stress tolerance was associated with a more extensive root system, a lower level of malondialdehyde, a nearly normal Na(+)/K(+) ratio, a higher level of proline and a kinetically accelerated induction of SOD and CAT activities observed in the transgenic plants during drought and/or salt stress, indicating that an enhanced ROS scavenging capability might play a significant role in the acquired tolerance to the abiotic stress.

Analysis of probenazole-responsiveness of rice RPR1 upstream fragments.

RPR1 (rice probenazole-responsive) is a rice gene, the expression of which is responsive to probenazole (PBZ), a synthetic compound that may act as a plant defense activator. It has been shown that RPR1 gene may be involved in disease resistance responses. In this study, a series of amplified fragments from the rice RPR1 promoter region, including 2,416 bp, 1,574 bp, 819 bp, 568 bp and 208 bp fragments upstream to the ATG translation start site, were prepared and linked to the coding region of beta-glucuronidase (GUS) gene. Analysis of GUS gene transient expression in rice calli demonstrated that the 568 bp fragment was sufficient for probenazole responsiveness. Analysis of GUS gene stable expression in Arabidopsis thaliana indicated that the 2,416 bp and 1,574 bp fragments drove GUS expression only in shoot apical meristem and petiole. Identification of these PBZ-responsive fragments provides a basis on which PBZ-inducible gene regulatory systems can be constructed for experimental analysis of gene expression and for field application.

Repression of AtCLH1 expression results in a decrease in the ratio of chlorophyll a/b but doesnot affect the rate of chlorophyll degradation during leaf senescence.

To explore the possible regulatory role of chlorophyllase (Chlase) in chlorophyll (Chl) degradation during leaf senescence, RNAi Arabidopsis (Arabidopsis thaliana) plants were constructed to repress the expressions of AtCLH1 and/or AtCLH2. Transcript levels of AtCLH1 and/or AtCLH2 were dramatically lowered and Chlase activity was correspondingly inhibited, but the Chl degradation kinetics was not affected in the RNAi plants. Results of further analysis indicated that the Chl a/b ratio decreased in AtCLH1 RNAi lines, in comparison with the increasing Chl a/b ratio in the wide type during leaf senescence. In addition, an induced Chlase activity was consistently detected at the initial stage of senescence in all the plants examined. In contrast, transcript levels of both AtCLH1 and AtCLH2 decreased dramatically upon the initiation of senescence in both the wide-type and the RNAi plants. Interestingly, compared with the wide type, lower but still significant transcript levels of the RNAi targeted Chlase gene(s) were sustained during the whole period of dark incubation in all the three RNAi lines examined, indicating the functioning of some compensatively regulating mechanism. Based on these results, along with related reports, we conclude that Chlase might be required at the initial stage of leaf senescence, quite likely playing a role in converting Chl b to a.

Characterization of a novel developmentally retarded mutant (drm1) associated with the autonomous flowering pathway in Arabidopsis.

A developmentally retarded mutant (drm1) was identified from ethyl methanesulfonate (EMS)-mutagenized M2 seeds in Columbia (Col-0) genetic background. The drm1 flowers 109 d after sowing, with a whole life cycle of about 160 d. It also shows a pleiotropic phenotype, e.g., slow germination and lower germination rate, lower growth rate, curling leaves and abnormal floral organs. The drm1 mutation was a single recessive nuclear mutation, which was mapped to the bottom of chromosome 5 and located within a region of 20-30 kb around MXK3.1. There have been no mutants with similar phenotypes reported in the literature, suggesting that DRM1 is a novel flowering promoting locus. The findings that the drm1 flowered lately under all photoperiod conditions and its late flowering phenotype was significantly restored by vernalization treatment suggest that the drm1 is a typical late flowering mutant and most likely associated with the autonomous flowering pathway. The conclusion was further confirmed by the revelation that the transcript level of FLC was constantly upregulated in the drm1 at all the developmental phases examined, except for a very early stage. Moreover, the transcript levels of two other important repressors, EMF and TFL1, were also upregulated in the drm1, implying that the two repressors, along with FLC, seems to act in parallel pathways in the drm1 to regulate flowering as well as other aspects of floral development in a negatively additive way. This helps to explain why the drm1 exhibits a much more severe late-flowering phenotype than most late-flowering mutants reported. It also implies that the DRM1 might act upstream of these repressors.

Tetraenol, a novel sesquiterpenoid from the relict plant Tetraena mongolica in China.

A novel furansesquiterpenoid, tetraenol, was isolated from a relict shrub plant, Tetraena mongolica, collected from the northern desert of the Ningxia Hui Autonomous Region. The structure of the new compound was elucidated on the basis of spectroscopic analysis.

[Screening for lifespan-extension mutants with paraquat in Arabidopsis].

Genetic analyses of lifespan in model animals have revealed that extended lifespans are closely associated to increased resistance to oxidative stress. In the model plant Arabidopsis, late-flowering mutants are also found to be more tolerant to oxidative stress. However, Arabidopsis mutants with extended lifespans are poorly studied so far. In this study, a screening system for mutants with extended lifespans in Arabidopsis was established using paraquat, a potent herbicide that exacerbates O2-. radical production. The relationship between lifespan and resistance to oxidative stress was examined with one of the mutants, SFNA-9-4. Compared to that of wild type, the lifespan of SFNA-9-4 is significantly extended, and its resistance to oxidative stress is also significantly elevated. These results suggest that, as in Drosophila, paraquat can also be used to screen for mutants with extended lifespans in Arabidopsis.