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Glial cells, consisting of astrocytes, oligodendrocyte lineage cells, and microglia, account for >50% of the total number of cells in the mammalian brain. They play key roles in the modulation of various brain activities under physiological and pathological conditions. Although the typical morphological features and characteristic functions of these cells are well described, the organization of interconnections of the different glial cell populations and their impact on the healthy and diseased brain is not completely understood. Understanding these processes remains a profound challenge. Accumulating evidence suggests that glial cells can form highly complex interconnections with each other. The astroglial network has been well described. Oligodendrocytes and microglia may also contribute to the formation of glial networks under various circumstances. In this review, we discuss the structure and function of glial networks and their pathological relevance to central nervous system diseases. We also highlight opportunities for future research on the glial connectome.
Subject(s)
Animals , Neuroglia/physiology , Neurons/physiology , Astrocytes , Microglia/physiology , Oligodendroglia , MammalsABSTRACT
Accurate methods for identifying pelvic lymph node metastasis (LNM) of prostate cancer (PCa) prior to surgery are still lacking. We aimed to investigate the predictive value of peripheral monocyte count (PMC) for LNM of PCa in this study. Two hundred and ninety-eight patients from three centers were divided into a training set (n = 125) and a validation set (n = 173). In the training set, the independent predictors of LNM were analyzed using univariate and multivariate logistic regression analyses, and the optimal cutoff value was calculated by the receiver operating characteristic (ROC) curve. The sensitivity and specificity of the optimal cutoff were authenticated in the validation cohort. Finally, a nomogram based on the PMC was constructed for predicting LNM. Multivariate analyses of the training cohort demonstrated that clinical T stage, preoperative Gleason score, and PMC were independent risk factors for LNM. The subsequent ROC analysis showed that the optimal cutoff value of PMC for diagnosing LNM was 0.405 × 109 l
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Objective To study the differences of the phenoloxidase (PO) relative activity among ribbed shelled Oncomelania hupensis, smooth shelled O. hupensis and Cipangopaludina chinensis. Methods The crude PO fluid was extracted from the soft tissue of O. hupensis and C. chinensis by homogenation and centrifugation. The PO activity was detected with catechol as the substrate in the reaction systems. Results The PO relative activities in the ribbed shelled O. hupensis, smooth shelled O. hupensis and C. chinensis were (25.72 ± 2.27), (14.56 ± 1.24) U / mL and (13.72 ± 1.06) U / mL. The PO relative activity in the smooth shelled O. hupensis was higher than that in the ribbed shelled O. hupensis (q = 21.46, P < 0.05) and C. chinensis (q = 12.00, P < 0.05), while the difference between the PO relative activities of the latter two was not statistically significant (q = 1.62, P > 0.05) . Conclusion There is a difference in the relative PO activity between O. hupensis and C. chinensis, which may be related to the living environment of snails.
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Cytochrome P450 family is a kind of biocatalyst widely existing in nature. It has many functions such as catalyzing the biosynthesis of plant secondary metabolites and regulating phytoremediation. Based on the analysis of proteome data of Tripterygium wilfordii,the CYP450 gene of T. wilfordii was preliminarily analyzed and predicted by various bioinformatics methods. The results showed that after the expression of T. wilfordii suspension cells was induced by methyl jasmonate,the proteomic data of T. wilfordii were obtained and analyzed,and 10 CYP450 proteins of T. wilfordii were finally screened out. By analyzing the phylogenetic tree constructed with CYP450 gene of Arabidopsis family,the 10 CYP450 proteins were clustered into 6 different CYP450 families. The physical and chemical properties of CYP450 proteins in different families were different. The secondary structure of CYP450 proteins was mainly composed of irregular curls. Eight subcellular localization results of CYP450 proteins were chloroplasts and the rest were plastids. Subsequently,the conserved domains( heme active sites) shared by CYP450 genes were found by analyzing the results of multiple sequence alignment. Finally,by analyzing the transcriptome data of T. wilfordii,the expression distribution of T. wilfordii in different tissues was preliminarily confirmed,which verified its correlation with the biosynthesis of active components of T. wilfordii,and provided important genetic resources for the analysis of biosynthesis pathway of active components of T. wilfordii.
Subject(s)
Computational Biology , Cytochrome P-450 Enzyme System , Chemistry , Phylogeny , Plant Proteins , Chemistry , Proteomics , Tissue Distribution , TripterygiumABSTRACT
Objective To study the differences of the phenoloxidase (PO) relative activity among ribbed shelled Oncomelania hupensis, smooth shelled O. hupensis and Cipangopaludina chinensis. Methods The crude PO fluid was extracted from the soft tissue of O. hupensis and C. chinensis by homogenation and centrifugation. The PO activity was detected with catechol as the substrate in the reaction systems. Results The PO relative activities in the ribbed shelled O. hupensis, smooth shelled O. hupensis and C. chinensis were (25.72 ± 2.27), (14.56 ± 1.24) U / mL and (13.72 ± 1.06) U / mL. The PO relative activity in the smooth shelled O. hupensis was higher than that in the ribbed shelled O. hupensis (q = 21.46, P < 0.05) and C. chinensis (q = 12.00, P < 0.05), while the difference between the PO relative activities of the latter two was not statistically significant (q = 1.62, P > 0.05) . Conclusion There is a difference in the relative PO activity between O. hupensis and C. chinensis, which may be related to the living environment of snails.
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Tripterygium wilfordii 3-hydroxy-3-methylglutaryl coenzyme-A reductase (TwHMGR) is an important regulation site in terpenoids metabolic pathway in cytoplasm which is the first speed limit enzyme of MVA pathway. In order to investigate the effects of TwHMGR on the biosynthesis of triptolide and celastrol in Tripterygium wilfordii, the overexpression of TwHMGR (OE-HMGR) was studied in this paper. We cloned the full-length of TwHMGR to construct overexpression vector by Gateway technology then delivered the expression vector into Tripterygium wilfordii suspension cells by gene gun. qRT-PCR was used to detect the expression of TwHMGR:the expression of TwHMGR was increased to 1.75 folds over the control group (empty vector:pH7WG2D) in the overexpression group. The accumulation of triptolide and celastrol in the suspension cells of Tripterygium wilfordii was detected by UPLC, revealing that:the contents of triptolide and celastrol were increased to 163.93% and 190.04% of over the control group in the overexpression group. Based on these findings, the positive effect on the accumulation of active terpenoids, triptolide and celastrol in Tripterygium wilfordii was found and the results laid a foundation of the synthetic biology research on important active terpenoids in Tripterygium wilfordii.
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Objective: To clone the full-length cDNA of phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase in Tripterygium wilfordii (TwPTEN) and predicted its biological functions. Methods: The specific primers were designed based on the transcriptome data of Tripterygium wilfordii, TwPTEN gene was cloned by Polymerase Chain Reaction. The full-length cDNA of TwPTEN was then analyzed by a series of bioinformatics analysis. Results: It was showed that the full length of TwPTEN cDNA was 2 247 bp encoding 614 amino acids. The theoretical isoelectric point was 5.84 and the molecular weight was about 66 900. Sequence alignment and phylogenetic analysis demonstrated that TwPTEN had relative close relationship to PTEN from other species. Differential expression analysis revealed that the relative expression level of TwPTEN increased significantly after being induced by methyl jasmonate (MeJA). It indicated that the TwPTEN gene was relative to the biosynthesis of secondary metabolites. Conclusion: PTEN gene is firstly cloned from T. wilfordii, which provides a basis for further studying the functions of TwPTEN.
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In this study, we cloned a monoterpene synthases, TwMS from Tripterygium wilfordii suspension cells. TwMS gene contained a 1 797 bp open reading frame (ORF), encoding a polypeptide of 579 amino acids, which deduced isoelectric point (pI) was 6.10 and the calculated molecular weight was 69.75 kDa. Bioinformation analysis showed that the sequence of TwMS was consistent with the feature of monoterpene synthases. Differential expression analysis revealed that the relative expression level of TwMS increased significantly after being induced by methyl jasmonate (MeJA). The highest expression level occurred at 24 h. TwMS protein was successfully expressed in Escherichia coli BL21 (DE3), which laid the foundation for identifying the function of T. wilfordii monoterpene synthases.
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Kaurenoic acid oxidase involved in biosynthesis pathway of gibberellin. According to the transcriptome database, the specific primers were designed and used in cloning the full-length cDNA of TwKAO, the bioinformatic analysis of the sequence was performed. The qRT-PCR were used to detect the expression level of TwKAO after MeJA treatment.The full-length cDNA of the TwKAO was 1 874 bp encoding a polypeptide of 487 amino acids.The calculate molecular weight was about 56.02 kDa,and the theoretical isoelectric point (pI) was 8.89. The relative expression level of TwKAO was deduced by MeJA and reached the highest at 12 h after the treatment.Plant tissue expression analysis indicated that, TwKAO expressed the highest in leaves,while lowest in roots.For the first time, we cloned and analyzed the expression characteristics of TwKAO, which laid a foundation for deep analysis of growing development and terpenoid secondary metabolites in T. wilfordii.
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<p><b>OBJECTIVE</b>To investigate the molecular mechanism of haemophilia B caused by the novel mutation of Arg327Ile (R327I) in FIX gene.</p><p><b>METHODS</b>The R327I, R327Ala(A), R327Lys(K), R327Asn(N) and a replacement mutant (FIXβFVII), in which FIX β strand 324-329 was replaced by that of FVII 298-303, expression plasmids were constructed with site-directed mutagenesis method based on the wild-type (WT) FIX expression plasmid. The HEK293 cell was transiently transfected, then the activity of FIX (FIX:C) was assayed by one stage method in the conditioned medium, while the FIX:Ag in both the conditioned media and the cell lysates was measured by ELISA. The molecular weight and the semi-quantity of expressed FIX were analyzed by Western blot. Fluorescent protein expression plasmid was constructed to investigate the synthesis and secretion of the FIX R327I mutation in the viable cells.</p><p><b>RESULTS</b>FIX:C of the R327I mutant protein was 4.49% of the level of the WT in the conditioned medium, and the FIX:Ag of the R327I mutant protein in the conditioned medium and the cell lysates was 31.02% and 129.29% compared to that of WT, respectively. The mutation was characterized as cross-reaction material reduced (CRMR). The viable cell fluorescent assays showed that the R327I protein was more in both the viable cells and in lysosome than that of WT. The FIX:C of the R327A, R327K, R327N and FIXβFVII mutants was reduced compared to that of WT, the reduction of FIX:C of FIXβFVII was the most significantly amount among all the mutants in medium. FIX:Ag of all the mutants in the medium, except that the R327K increased, was reduced. The result of Western blot showed that the molecular weight of R327I protein was the same as that of WT, but the amount of the protein was much less compared with WT in the conditioned medium.</p><p><b>CONCLUSION</b>The abnormal synthesis and secretion as well as the abnormal function of the R327I mutant protein causes haemophilia B. The residue of R327 as well as the β strand domain of R327 located play important roles of the specific function of FIX.</p>
Subject(s)
Humans , Factor IX , Genetics , HEK293 Cells , Hemophilia B , Genetics , Pathology , Mutagenesis, Site-Directed , Mutation , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To evaluate the contribution of platelet and leukocyte activation in pathogenesis primary pulmonary hypertension (PPH).</p><p><b>METHODS</b>Pulmonary hypertension was induced by subcutaneous injection of 2% monocrotaline (MCT) in male Prague-Dawley (SD) rats. Blood samples were collected at the third week after MCT injection, and flow cytometry was used to determine the fibrinogen-binding platelet, CD11b expression on leukocyte and platelet-leukocyte aggregation.</p><p><b>RESULT</b>Three weeks after MCT injection, rats exhibited higher right ventricular systolic pressure (RVSP) and mean pulmonary artery pressure(mPAP), as compared with controls. MCT induced vascular remodeling characterized by vascular medial wall thickening in pulmonary muscular arteries. The ratio of platelets fibrinogen binding was increased in rats 3 weeks after MCT injection than that of control group[(4.08 +/-1.59)% compared with (1.45 +/- 0.61)%, P<0.01]. CD11b expression in monocytes and neutrophils, but not in lymphocytes was increased significantly 3 weeks after MCT injection (P <0.01). Platelet-neutrophil aggregations increased in MCT injected rats as compared with controls (P <0.01).</p><p><b>CONCLUSION</b>Rats of PPH model demonstrate enhanced circulating platelet and leukocyte activation, which may contribute to the pathogenesis of PPH.</p>