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Lei Ding

Publications and source records attributed to Lei Ding.

At least 37 records · Page 2Linked to original sources

Short-term lineage analysis of dorsally derived Olig3 cells in the developing spinal cord.

We examined the migration and differentiation of cells expressing Olig3, a basic helix-loop-helix transcriptional factor, in the developing spinal cord. Distribution of Olig3 lineage cells was demonstrated with in situ hybridization and X-gal staining in an Olig3-lacZ knock-in mouse. Olig3-positive cells first appeared in the dorsal spinal cord, except for the roof plate. Some of the dorsal Olig3 lineage cells co-expressed Islet1/2, Math1, or Brn3a, markers for dorsal interneuron. LacZ-positive cells were observed in the ventral-most part of the E10.5 spinal cord, suggesting that some dorsal Olig3 lineage cells migrate into the ventral-most part by E10.5. Ventral-ward migration of dorsal cells and contribution to commissural interneurons were substantiated by electroporation of EGFP expression plasmid in the dorsal spinal cord of chick embryo. Dorsal midline cells were also LacZ-positive during development. These findings suggest that dorsal Olig3 cells contribute to dorsal midline cells and commissural interneurons at intermediate and ventral levels.

Animals↗

Estimation of the cortical connectivity by high-resolution EEG and structural equation modeling: simulations and application to finger tapping data.

Today, the concept of brain connectivity plays a central role in the neuroscience. While functional connectivity is defined as the temporal coherence between the activities of different brain areas, the effective connectivity is defined as the simplest brain circuit that would produce the same temporal relationship as observed experimentally between cortical sites. The most used method to estimate effective connectivity in neuroscience is the structural equation modeling (SEM), typically used on data related to the brain hemodynamic behavior. However, the use of hemodynamic measures limits the temporal resolution on which the brain process can be followed. The present research proposes the use of the SEM approach on the cortical waveforms estimated from the high-resolution EEG data, which exhibits a good spatial resolution and a higher temporal resolution than hemodynamic measures. We performed a simulation study, in which different main factors were systematically manipulated in the generation of test signals, and the errors in the estimated connectivity were evaluated by the analysis of variance (ANOVA). Such factors were the signal-to-noise ratio and the duration of the simulated cortical activity. Since SEM technique is based on the use of a model formulated on the basis of anatomical and physiological constraints, different experimental conditions were analyzed, in order to evaluate the effect of errors made in the a priori model formulation on its performances. The feasibility of the proposed approach has been shown in a human study using high-resolution EEG recordings related to finger tapping movements.

Algorithms↗

[Treating high strength antibiotic wastewater by micro-aerobic hydrolysis and acidification process].

The effect of micro-aerobic hydrolysis and acidification to high strength antibiotic wastewater treatment is studied. The results demonstrate that micro oxygen enhanced the physiological metabolizability of facultative hydrolytic and acidogenic bacteria, and aerating stirring improved the hydraulic condition. Degree of acidification (AD) and volatile fatty acid (VFA) in the effluent reached 58.64% and 4825 mg/L with the shortest HRT 10 h and the maximal OLR 20 kg/(m3 x d), respectively. Wastewater biodegradability was improved greatly and rising BOD5/COD was about 17%, which offered good substrate for post aerobic treatment. The effluent quality was relative stable with the fluctuant influent, and COD and SS were 7000-8000 mg/L and 150-300 mg/L, respectively, while the removal efficiencies of COD and SS were 15%-30% and 90%-95%. The changement of VFA lagged behind that of AD in the effluent, and AD would represent the effect of hydrolysis and acidification process more properly. The sludge bed in the bottom of the reactor was the main reaction field for VFA production, and the height of the reactor for stable VFA production increased with the increase of OLR. The filling field mainly aimed to entrap SS in the effluent and benefited little to VFA production. Sludge in the reactor was mainly little sludge particles with the size of 0.5-1.0 mm and flocculent sludge.

Anti-Bacterial Agents↗

Involvement of extracellular signal-regulated kinase/mitogen-activated protein kinase pathway in multidrug resistance induced by HBx in hepatoma cell line.

AIM: To investigate the molecular mechanism of the influence of HBx protein on multidrug resistance associated genes: multidrug resistance 1 (MDR-1), multidrug related protein (MRP-1), lung resistance related protein (LRP) in hepatoma cells and the potential role of extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) pathway in this process. METHODS: A cell model stably expressing the HBx protein was established by liposome-mediated transfection of HBx gene into HepG2 cell line. The expression of multidrug resistance associated genes and proteins was detected by RT-PCR and Western blot. AnnexinV-FITC/PI assay was used to confirm the multidrug resistance (MDR) phenotype of transfected cells by fluorescence cytometry (FACS). The ERK/MAPK pathway activation was measured by Western blot through comparing the ratio of phosphorylation of ERK/MAPK to total ERK/MAPK protein. After treated with the ERK/MAPK pathway inhibitor U0126, the HBx-expressing cells were harvested. Then RT-PCR, Western blot and FACS were used to analyze the alterations in the expression of multidrug resistance associated genes and the MDR phenotype after exposure. RESULTS: Compared with the control group, the transfected cells showed a higher expression of MDR associated genes and proteins. Marked elevations in MDR-1 (64.3%), MRP-1 (87.5%) and LRP (90.8%) were observed in the transfected cells (P<0.05). RT-PCR revealed that the over-expression of MDR associated proteins was due to amplification of such genes (MDR1 2.9 fold, MRP1 1.67 fold, LRP1.95 fold). Furthermore, we found that the ERK/MAPK activity was remarkably high in the HBx-expressing cells. The activation of ERK/MAPK, as measured by the ratio of phosphorylated ERK bands normalized to the total ERK bands, was increased by 2.3-fold in HBx-transfected cells compared with cells transfected with the empty vector. After treated with the ERK/MAPK pathway inhibitor, the level of MDR associated genes and proteins in the transfected cells decreased to some extent. Compared with controls, a significant decrease in MDR-1 mRNA (53.3%), MRP-1 mRNA (59.7%) as well as LRP mRNA (56.4%) was observed in the U0126 treated transfected cells after 12 h. Western blot also demonstrated that the protein expression of these MDR associated genes slightly reduced after treated with U0126 for 12 h (MDR-1 40.1%, MRP-1 29.4%, LRP35.7%). This change was accompanied with the rise of cell apoptosis ratio confirmed by Annexin V-PI detection. The apoptosis index of U0126-treated cells increased by 1.28 fold, compared with that of transfected cells. Obviously, the MDR phenotype of these cells was obviously related with increased activities of the ERK/MAPK pathway. CONCLUSION: HBx protein might be one of the causes for the occurrence of MDR in HCC, and ERK/MAPK pathway might be involved in this change.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Improved preservation of X-gal reaction product for electron microscopy using hydroxypropyl methacrylate.

In lineage tracing analysis, the beta-galactosidase (beta-gal) gene is a commonly used as a reporter gene because it is relatively stable and highly sensitive in histochemical detection using 5-bromo-4-chloro-3-indolyl-beta-d-galactoside (X-gal). Clear determination of the types and characteristics of labeled cells requires transmission electron microscopic (TEM) examination of their morphology. X-gal staining, which involves the precipitate formed by the reaction between beta-gal and X-gal, is usually recognized as a light blue or green reaction product on light microscopic (LM) examination. However, the standard protocol for TEM preparation weakens the intensity of or results in the loss of X-gal reaction product at the step of substitution of ethanol with Epon using propylene oxide. To solve this problem, we show that hydroxypropyl methacrylate achieves good preservation of X-gal reaction products. The protocol presented here appears to be useful for lineage determination by TEM of all types of X-gal-stained tissues.

Animals↗

Motor imagery classification by means of source analysis for brain-computer interface applications.

We report a pilot study of performing classification of motor imagery for brain-computer interface applications, by means of source analysis of scalp-recorded EEGs. Independent component analysis (ICA) was used as a spatio-temporal filter extracting signal components relevant to left or right motor imagery (MI) tasks. Source analysis methods including equivalent dipole analysis and cortical current density imaging were applied to reconstruct equivalent neural sources corresponding to MI, and classification was performed based on the inverse solutions. The classification was considered correct if the equivalent source was found over the motor cortex in the corresponding hemisphere. A classification rate of about 80% was achieved in the human subject studied using both the equivalent dipole analysis and the cortical current density imaging analysis. The present promising results suggest that the source analysis approach could manifest a clearer picture on the cortical activity, and thus facilitate the classification of MI tasks from scalp EEGs.

Algorithms↗

Tetraspanin protein CD9 is a novel paranodal component regulating paranodal junctional formation.

The axoglial paranodal junction is essential for the proper localization of ion channels around the node of Ranvier. The integrity of this junction is important for nerve conduction. Although recent studies have made significant progress in understanding the molecular composition of the paranodal junction, it is not known how these membrane components are distributed to the appropriate sites and interact with each other. Here we show that CD9, a member of the tetraspanin family, is present at the paranode. CD9 is concentrated in the paranode as myelination proceeds, but CD9 clusters become diffuse, associated with disruption of the paranode, in cerebroside sulfotransferase-deficient mice. Immunohistochemical and Western blot analysis showed that CD9 is distributed predominantly in the PNS. Ablation of CD9 in mutant mice disrupts junctional attachment at the paranode and alters the paranodal components contactin-associated protein (also known as Paranodin) and neurofascin 155, although the frequency of such abnormalities varies among individuals and individual axons even in the same mouse. Electron micrographs demonstrated that compact myelin sheaths were also affected in the PNS. Therefore, CD9 is a myelin protein important for the formation of paranodal junctions. CD9 also plays a role in the formation of compact myelin in the PNS.

Animals↗

Estimation of the effective and functional human cortical connectivity with structural equation modeling and directed transfer function applied to high-resolution EEG.

Different brain imaging devices are presently available to provide images of the human functional cortical activity, based on hemodynamic, metabolic or electromagnetic measurements. However, static images of brain regions activated during particular tasks do not convey the information of how these regions are interconnected. The concept of brain connectivity plays a central role in the neuroscience, and different definitions of connectivity, functional and effective, have been adopted in literature. While the functional connectivity is defined as the temporal coherence among the activities of different brain areas, the effective connectivity is defined as the simplest brain circuit that would produce the same temporal relationship as observed experimentally among cortical sites. The structural equation modeling (SEM) is the most used method to estimate effective connectivity in neuroscience, and its typical application is on data related to brain hemodynamic behavior tested by functional magnetic resonance imaging (fMRI), whereas the directed transfer function (DTF) method is a frequency-domain approach based on both a multivariate autoregressive (MVAR) modeling of time series and on the concept of Granger causality. This study presents advanced methods for the estimation of cortical connectivity by applying SEM and DTF on the cortical signals estimated from high-resolution electroencephalography (EEG) recordings, since these signals exhibit a higher spatial resolution than conventional cerebral electromagnetic measures. To estimate correctly the cortical signals, we used a subject's multicompartment head model (scalp, skull, dura mater, cortex) constructed from individual MRI, a distributed source model and a regularized linear inverse source estimates of cortical current density. Before the application of SEM and DTF methodology to the cortical waveforms estimated from high-resolution EEG data, we performed a simulation study, in which different main factors (signal-to-noise ratio, SNR, and simulated cortical activity duration, LENGTH) were systematically manipulated in the generation of test signals, and the errors in the estimated connectivity were evaluated by the analysis of variance (ANOVA). The statistical analysis returned that during simulations, both SEM and DTF estimators were able to correctly estimate the imposed connectivity patterns under reasonable operative conditions, that is, when data exhibit an SNR of at least 3 and a LENGTH of at least 75 s of nonconsecutive EEG recordings at 64 Hz of sampling rate. Hence, effective and functional connectivity patterns of cortical activity can be effectively estimated under general conditions met in any practical EEG recordings, by combining high-resolution EEG techniques and linear inverse estimation with SEM or DTF methods. We conclude that the estimation of cortical connectivity can be performed not only with hemodynamic measurements, but also with EEG signals treated with advanced computational techniques.

Adult↗

Mycophenolate mofetil combined with prednisone for diffuse proliferative lupus nephritis: a histopathological study.

Mycophenolate mofetil (MMF) is a potential new treatment for diffuse proliferative lupus nephritis. This study examines the clinical and histopathological effects, and potential mechanisms, of combination MMF/prednisone therapy in diffuse proliferative lupus nephritis. Nine patients with diffuse proliferative lupus nephritis confirmed by renal biopsy received MMF/prednisone for six months when repeat biopsies were performed. Clinical and histopathological parameters of activity and chronicity were studied. Collagens were detected by Sirus red staining; leucocyte phenotype, osteopontin (OPN), fibrinectin (FN), alpha-smooth muscle actin (alpha-SMA) and TGF-beta1 were detected by immunohistochemistry. The changes of clinical and histopathologic parameters were assessed and compared to histopathologic indicators. Eight of the nine patients achieved clinical remission; renal function deteriorated in one. Histopathological activity indices reduced significantly (9.56 +/- 2.83 versus 5.22 +/- 1.86, P < 0.01); however, the chronicity indices did not change (3.56 +/- 1.42 versus 3.22 +/- 1.20). T-cell and monocyte/macrophage infiltration. OPN expression and the percentage of proliferative cells in both glomerulus and tubulo-interstitium decreased significantly. Other features of chronic lesions, except for glomerular collagen deposition, did not change. In conclusion, MMF/prednisone therapy was effective for our patients with proliferative lupus nephritis. The active inflammatory lesions could be ameliorated through reduction of lymphocyte and monocyte/macrophage infiltration, inhibition of cell proliferation and downregulation of adhesive molecules. However, the chronic fibrotic lesions could not be significantly reduced.

Actins↗

[Screening for low-density lipoprotein receptor gene mutations in familial hypercholesterolemia Chinese].

OBJECTIVE: To screen the mutations of low-density lipoprotein receptor (LDLR) gene in Chinese familial hypercholesterolemia (FH) patients. METHODS: 7 patients with clinical phenotype of homozygous FH and their parents were investigated for mutations in all the eighteen exons of LDLR gene. Screening was carried out using PCR-SSCP and direct DNA sequencing and LDLR gene mutation database was searched to identify the alteration. In addition, the apolipoprotein B gene (apo B) was screened for known mutations (R3500Q) that caused familial defective apo B100 (FDB) with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Nine mutations in the LDLR gene were identified in the FH patients. All the mutations except C255R have not been published in the LDLR gene mutation database. No mutation of apo B100 (R3500Q) was observed. CONCLUSIONS: Chinese FH patients may have specific spectrum and regional difference of LDLR gene mutations. Apolipoprotein B-100 gene mutation might not be the main cause of hypercholesterolemia patients in China.

Adolescent↗

[Colleagues Association: A medical organization in modern Japan].

Colleagues Association was established in Tokyo, Japan in June, 1902 as a professional medical group aiming at the spread of advanced Japanese medical science in China and to other neighboring countries. Until February 1946, it carried out many medical activities such as opening hospitals and publishing journals for over four decades. It flaunted the banner of medical spread and exchange, but on the other hand, it provided Japanese army with medical service and facilities and resisted European and American medical activities after being supervised by the Japanese government with the aggression against China. So it changed into a medical organization to assist the Japanese government to invade Asia.

China↗

[Expression and clinical significance of HIF-1a protein in hepatocellular carcinoma tissues].

OBJECTIVE: To investigate the expression and clinical significance of HIF-1a protein in hepatocellular carcinoma (HCC) tissues. METHODS: Immunohistochemistry (IHC), Western blotting and RT-PCR techniques were used to detect the expression of the HIF-1a gene protein in 35 HCC, 26 cirrhotic and 15 normal liver tissues. Their relationship with the pathological characteristics of the tumors were also analyzed. RESULTS: The positive rates of HIF-1a expression in HCC tissues was 94%, which was similar to the positive rates of HIF-1a expression in liver cirrhosis tissues of 92%, but was higher than that in normal hepatic tissues of 7%, but the residual proliferatic hepatic trabeculae among the necrotic liver cells and the fibrotic tissues expressed HIF-1a strongly in comparison with the cirrhotic liver tissues. The expression intensity of HIF-1a protein of the cirrhotic liver tissues was stronger than that in HCC; the results by Western blotting and RT-PCR were in accordance to that by IHC. In addition, the expression intensity in HCC had a negative correlation in differentiation degree and a positive correlation to intrahepatic and extrahepatic metastases but no correlation was found between HIF-1a expression and the existence of portal vein tumor emboli, prognosis and the status of HBsAg. CONCLUSION: HIF-1 protein was expressed in HCC and cirrhotic liver tissues, and was only affected by the factor of hypoxia. The expression of HIF-1a protein is associated with the differentiation of the tumor and its intrahepatic and extrahepatic metastases but was not related to the existence of portal vein tumor emboli, prognosis and the status of HBsAg. This phenomenon may provide a new idea for the treatment of liver cancer.

Adolescent↗

Physiological and biochemical changes in summer and winter diapause and non-diapause pupae of the cabbage armyworm, Mamestra brassicae L. during long-term cold acclimation.

Previous studies have demonstrated that cold hardiness in the cabbage armyworm Mamestra brassicae is more developed in winter diapause pupae (WD) than in summer diapause pupae (SD) after short-term cold acclimation. In this study, we investigated specific physiological and biochemical differences between diapause and non-diapause pupae (ND) of M. brassicae after acclimation at 5 degrees C for up to 4 months. The period required for adult emergence in ND pupae at 20 degrees C was about 20.2 d and independent of the months acclimated. However, SD and WD pupae reached this level only after 1- and 4-month acclimation, respectively. Levels of respiration (O(2) consumption and CO(2) release) in ND(0) (the subscript indicates the months of acclimation at 5 degrees C) was eightfold higher than in WD(0) and SD(0), decreased sharply in ND(1) and reached diapause pupae (WD and SD) levels in ND(2). Maximal cold hardiness developed during cold acclimation was found in WD(3), SD(2) and ND(2) pupae. During cold acclimation, glycogen content remained at higher levels in WD and SD than in ND. Lipid content in ND decreased but remained high in SD and WD. These results suggest that diapause pupae show large amounts of metabolic reserves in the form of glycogen and lipid during diapause development. To our knowledge, this is the first report to compare the physiological and biochemical differences among summer and winter diapause pupae and non-diapause pupae in the same insect species during long-term cold acclimation.

Acclimatization↗

[Hepatitis B x protein activated vascular endothelial growth factor expression through hypoxia inducible factor-1 pathway].

OBJECTIVE: To investigate whether hepatitis B x protein (HBx) stimulates vascular endothelial growth factor (VEGF) through hypoxia inducible factor-1 (HIF-1 alpha) pathway. METHODS: Two plasmids including pIRES-EGFP-HBx and pTK-Hyg were co-transfected to a hepatocellular carcinoma cell line SMMC-7721. With fluorescence-positive and fluorescence-negative hygromycin-resistant colonies selected, expressions of VEGF and HIF-1 alpha in protein or/and mRNA level were detected. RESULTS: Fluorescence-positive cells were stably integrated with HBx, in which expression of HIF-1 alpha and VEGF were upregulated. Fluorescence-negative cells did not express HBx, VEGF or HIF-1 alpha. CONCLUSION: HBx can activate VEGF through HIF-1 alpha pathway.

Cell Line, Tumor↗

Drosophila Enhancer of zeste protein interacts with dSAP18.

The Drosophila Enhancer of zeste [E(z)] gene encodes a member of the Polycomb group of transcriptional repressors. Here we report evidence for direct physical interaction between E(Z) and dSAP18, which previously has been shown to interact with Drosophila GAGA factor and BICOID proteins. dSAP18 shares extensive sequence similarity with a human polypeptide originally identified as a subunit of the SIN3A-HDAC (switch-independent 3-histone deacetylase) co-repressor complex. Yeast two-hybrid and in vitro binding assays demonstrate direct E(Z)-dSAP18 interaction and show that dSAP18 is capable of interacting with itself. Co-immunoprecipitation experiments provide evidence for in vivo association of E(Z) and dSAP18. Gel filtration analysis of embryo nuclear extracts shows that dSAP18 is present in native protein complexes ranging from approximately 1100 to approximately 450 kDa in molecular mass. These studies provide support for a model in which dSAP18 contributes to the activities of multiple protein complexes, and potentially may mediate interactions between distinct proteins and/or protein complexes.

Animals↗

Seasonal changes in glycerol content and enzyme activities in overwintering larvae of the Shonai ecotype of the rice stem borer, Chilo suppressalis Walker.

Enzymes associated with glycogen metabolism and glycerol synthesis in larvae of the Shonai ecotype of the rice stem borer, Chilo suppressalis, were investigated over the winter in 2000-2001. Glycerol content was scarcely detected in September and October, rapidly increased in November and December, peaked in January, and then decreased. Glycogen was converted to glycerol over the winter until February, and glycerol was reconverted to glycogen in March. The trehalose content remained constant over the winter. The activities of enzymes associated with glycerol synthesis changed with the season. Glycerol accumulation was accomplished by activation of glycogen phosphorylase, inhibition of fructose-1,6-bisphosphatase and pyruvate kinase, and activation of enzymes associated with glycerol synthesis, mainly glyceraldehyde-3-phosphatase and polyol dehydrogenase with glyceraldehyde activity. These changes led to a diversion of triose phosphates into the pathway of glycerol synthesis. However, activities of the two initial enzymes of the hexose monophosphate shunt were not activated and remained relatively constant, but high during the period of active glycerol synthesis. Both decreasing temperature in the field and the transition from the diapause to the post-diapause state may be responsible for the changes in activities of enzymes associated with glycerol synthesis.

Animals↗

Diapause development and acclimation regulating enzymes associated with glycerol synthesis in the Shonai ecotype of the rice stem borer larva, Chilo suppressalis walker.

Overwintering larvae of the Shonai ecotype of the rice stem borer, Chilo suppressalis, enter diapause in early September and terminate diapause at the end of October. Cold acclimation at 0 degrees C did not influence glycerol, trehalose or glycogen content in larvae collected on 22 September. Acclimation at 0 degrees C increased the glycerol content and reduced the glycogen content significantly in larvae collected on 2 October and 22 November compared with acclimation at 15 degrees C. These results indicate that overwintering larvae at different phases of diapause development respond differently to the low temperature stimulus for glycerol synthesis. Thus, we evaluated the metabolic rearrangements associated with glycerol synthesis during diapause development and after temperature acclimation. Larvae collected on 2 October were acclimated at 15 degrees C for 15 and 60 days. Some of those acclimated at 15 degrees C were then moved to 0 degrees C for 15 days. The larvae acclimated at 15 degrees C for 15 days were in deep diapause and accumulated little glycerol, while larvae acclimated at 15 degrees C for 60 days were nearly ready to emerge from diapause and accumulated glycerol at 155.5 &mgr;mol/g. When larvae acclimated to 15 degrees C for 15 days were transferred to 0 degrees C, glycerol accumulation was stimulated to the same extent (ca 140 &mgr;mol/g) as it was in larvae that were acclimated to 15 degrees C for 60 days and then transferred to 0 degrees C. These results indicate that low temperature has a cumulative effect on glycerol production in larvae at different phases of diapause development. Glycerol accumulation was accomplished by activation of glycogen phosphorylase and inhibition of fructose-1,6-bisphosphatase, and activation of enzymes associated with glycerol synthesis, mainly glyceraldehyde-3-phosphatase and polyol dehydrogenase with glyceraldehyde activity.

Journal Article↗