Schwann-like adipose-derived stem cells as a promising therapeutic tool for peripheral nerve regeneration:effects of cholinergic stimulation

Schwann-like adipose-derived stem cells and nerve injury:Peripheral nerve injuries(PNIs)are a common clinical problem usually as a consequence of trauma.Despite optimal surgical management,PNI has a lifelong impact on function and wellbeing of the patient.The peripheral nervous system(PNS)has regenerative capability,in contrast to the central nervous system(CNS),and is dependent on the plasticity of the peripheral glia,Schwann cells(SCs).Despite this regenerative capability,PNI recovery of sensorial and motor function is always incomplete causing pain,cold intolerance,paralysis and impairment of activities of daily living.

Clinical significance of monocyte-derived dendritic cell activation in patients with acute coronary syndrome

Background: Acute coronary syndrome (ACS) is an amplified state of inflammation and immune reaction. Dendritic cells (DCs) expressing various Toll-like receptors (TLRs) have been observed in atherosclerotic lesions, however, the clinical significance of DCs in pathogenesis of ACS has not been completely investigated. Methods: Ten patients with ACS and 10 patients with stable angina pectoris (SAP) were enrolled in this study. Monocyte-derived DCs were generated from CD14+ cells by culturing with granulocyte macrophage colony-stimulating factor and interleukin (IL)-4 for 6 days. Expression of cell surface CD86 and CD83 were measured by flowcytometry. Expression of genes, including CD86, CD83, CCL19, CCR7, TLR2, TLR4, TLR5, TLR8, and TLR9, were measured by real-time PCR. Plasma IL-6 and tumor necrosis factor (TNF)-α levels were also measured. Results: The number of CD86+CD83+DCs in the ACS group was significantly higher than that in the SAP group (P P P +CD83+ cells and plasma levels of IL-6 (P = 0.88, P +CD83+ cells TNF-α levels (r = 0.78, P < 0.0001). Conclusions: These results demonstrated that mono-cyte-derived DCs are activated in patients with ACS, suggesting that activated DCs may play an important role in the pathogenesis of ACS.

Disrupted regulatory T cell homeostasis in inflammatorybowel diseases

In the gut, where billions of non-self-antigens from the food and the microbiota are present, the immune response must be tightly regulated to ensure both host protection against pathogenic microorganisms and the absence of immune-related pathologies. It has been well documented that regulatory T cells(Tregs) play a pivotal role in this context. Indeed, Tregs are able to prevent excessive inflammation, which can lead to the rupture of intestinal homeostasis observed in inflammatory bowel diseases(IBDs). Both the worldwide incidence and prevalence of such diseases have increased throughout the latter part of the 20^(th) century. Therefore, it is crucial to understand how Tregs suppress the colitogenic immune cells to establish new treatments for patients suffering from IBDs. In this review, we will first summarize the results obtained in animal model studies that highlight the importance of Tregs in maintaining intestinal homeostasis and describe the specific suppressive mechanisms involved. Next, our current knowledge about Tregs contribution to human IBDs will be reviewed, as well as the current therapeutic perspective on using Tregs for clinical IBD treatment and the challenges that remain to be resolved to ensure both the safety and effectiveness of these therapies in targeting this critical immune-regulatory cell population.

Tissue with high intelligence quotient Adipose-derived stem cells in neural regeneration

The aim of the present review is to highlight the possible neuroregenerative potential ol adipose-derived stem cells. The key property of stem cells is plasticity including self-renewal, multilineage differentiation, and migration, whereas the required property is transplantability. For a long time, embryonic stem cells were thought to be the only source of pluripotency, a dogma that has been challenged during the last decade. Today, an alternative option might be adipose-derived stem cells, as easily accessible, ethical and autologous cellular source. Recent knowledge of adipobiology increasingly recognizes that adipose tissue is the major endo- and paracrine organ of the human body. Likewise, numerous neuropetides, neurotrophic factors, neurotransmitters, hypothalamic and steroid hormones and their receptors are shared by adipose tissue and brain. Accordingly, the regenerative potential of neuroprotective factor-secreting adipose-derived stem cells is outlined. Whether the possible benefits of adipose stem cell-based therapy may be mediated via cell transdifferentiation and/or paracrine mechanisms remains to further be evaluated.

Genetic Structure of the Red-spotted Tokay Gecko, Gekko gecko(Linnaeus, 1758)(Squamata: Gekkonidae) from Mainland Southeast Asia

This study was performed to explore the genetic diversity and genetic structure of red-spotted tokay geckos(Gekko gecko) from 23 different geographical areas in Thailand, Lao PDR and Cambodia. The mitochondrial tRNAGln/tRNA-Met/partial NADH dehydrogenase subunit 2 from 166 specimens was amplified and sequenced. A total of 54 different haplotypes were found. Highly significant genetic differences occurred between populations from different localities. The haplotype network revealed six major haplogroups(G1 to G6) belonging to different clades(clade A–E). Clade D and clade E were newly observed in this study. Haplogroup G4(clade D) was a sympatric population with haplogroup G1(clade B). The populations from northern Thailand were divided into two distinct haplogroups separated by mountain range. Genetic structure and genetic differentiation of the tokay in Southeast Asia was related to the geographical region sampled, spatial distance and natural barriers. Our results indicate that red-spotted tokay geckos from mainland Southeast Asia are cryptically diverse. Morphological comparisons, in addition to an intensive genetic investigation covering the whole species range, are needed to clarify the systematic and population structure of this species group.

Genetic Differentiation of the Forest Crested Lizards,Calotes emma alticristatus Schmidt,1925 and C.emma emma Gray,1845(Squamata:Agamidae)Relating to Climate Zones in Thailand

In Thailand,the forest crested lizard,Calotes emma consists of two subspecies,C.emma alticristatus Schmidt,1925 and C.emma emma Gray,1845.This study was performed to determine the genetic diversity and differentiation of C.emma from 16 different localities throughout Thailand.A total of 116 samples were analyzed using the mitochondrial cytochrome c oxidase subunit 1(CO1).Of these,65 and 51 of C.e.alticristatus and C.e.emma were classified into 23(N1-N23)and 21(S1-S21)haplotypes,respectively.There was no shared haplotype between subspecies or between different populations within each subspecies.These haplotypes were classified into four(north-A to north-D)and three(south-A to south-C)haplogroups of C.e aliristaus and C.e.emma,respectively.Phylogenetic analyses retrieved four lineages(classified asⅠtoⅣ).LineagesⅠandⅡcontained the four haplogroups of C.e.alicristatus,whereas lineageⅡandⅣcontained three haplogroups of C.e.emma.These two subspecies live separately in different climate zones,ie.C.e.alticristatus is found in an equatorial winter dry climate,whereas C.e.emma inha bits areas with an equa torial monsoonal clima te.

Multilocus enzyme electrophoresis analysis of Echinostoma revolutum and Echinostoma malayanum(Trematoda:Echinostomatidae) isolated from Khon Kaen Province,Thailand

Objective:To explore the genetic variation and differentiation of 2 echinostomes from genus Echinostoma,i.e.Echinostoma revolutum(E.revolution) and Echinostoma malayanum(E. malayanum) from Khon Kaen Province,Thailand.Methods:These parasites were compared at 22 enzymes encoding a presumptive 30 loci by using multilocus enzyme electrophoresis (MEE) technique.Results:Twenty-two loci can be used as diagnostic markers to differentiate these 2 species.E.revolutum and E.malayanum had fixed genetic differences at 70%of loci, whereas both species had fixed genetic differences from the liver fluke,Opisthorchis viverrini at 91%of loci.Intraspecific variation within a population of E.revolutum was observed at 5 polymorphic loci.Conclusions:MEE is a powerful technique to investigate genetic variation and differentiation of E.revolutum and E.malayanum.

Genetic variation and phylogenetic relationship of Hypoderaeum conoideum(Bloch, 1782) Dietz, 1909(Trematoda: Echinostomatidae) inferred from nuclear and mitochondrial DNA sequences

Objective:To explore genetic variations of Hypoderaeum conoideum collected from domestic ducks from 12 different localities in Thailand and Lao PDR,as well as their phylogenetic relationship with American and European isolates.Methods:The nucleotide sequences of their nuclear ribosomal DNA(ITS),mitochondrial cytochrome c oxidase subunit 1(CO1),and NADH dehydrogenase subunit 1(ND1)were used to analyze genetic diversity indices.Results:We found relatively high levels of nucleotide polymorphism in ND1(4.02%),whereas moderate and low levels were observed in CO1(2.11%)and ITS(0.96%),respectively.Based on these polymorphisms,the 20 ND1,12 CO1,and 18 ITS haplotypes were classified,and several common haplotypes were observed in all samples.At least three major lineages,namely American,European and Asian lineages,have been classified by phylogenetic analyses based on ND1 sequences.Conclusions:Our report demonstrates that the ND1 gene is the most suitable genetic marker to explore genetic variation and phylogenetic relationship of Hypoderaeum conoideum.However,a combination of all loci for ND1,CO1 and ITS would be of great value toward further genetic investigation of this endemic worldwide parasite.Thus,comprehensive molecular genetic analyses of Hypoderaeum conoideum from its worldwide distribution is needed to further understanding of the evolutionary and systematic relationships of this parasite.

Genetic Diversity and Population Structure of the Oriental Garden Lizard,Calotes versicolor Daudin,1802(Squamata:Agamidae)along the Mekong River in Thailand and Lao PDR

Calotes versicolor Daudin,1802,is geographically widespread along the Mekong River basin.The Mekong River is play important role as a significant natural barrier to several terrestrial animals living on different sides.This study aims to analyze the genetic diversity and population structure of C.versicolor populations collected from different sides of Mekong River using mitochondrial cytochrome c oxidase subunit 1(CO1)sequences.We obtained sequences of 200 individuals from 18 sampling localities from left and right sides of the Mekong River in Lao PDR and Thailand respectively.Overall,91 haplotypes were detected,which reflect high levels of genetic diversity in this species at the study areas.Haplotype network and phylogenetic analyses revealed that there were six major lineages(lineage C–lineage H)of C.versicolor populations within the Mekong River,whereas lineages A and B have previously been found from China and Vietnam.The genetic distance among C.versicolor was significantly related to spatial distance,however,the Mekong River had no significant effect on genetic distance.Our findings,together with previous studies,suggests that C.versicolor in Asia is a species complex with other cryptic lineages being likely but there is a need for further exploration.Thus,comprehensive genetic,biological and ecological studies of C.versicolor should be conducted throughout its entire distribution range.

In Vitro Evaluation of Two Tissue Substitutes for Gingival Augmentation

Three-dimensional collagen matrices of porcine origin are being used as substitutes for soft tissue grafts in periodontal plastic surgery in search of aesthetic and natural results. This in vitro study aimed to compare Fibro-Gide® (GeistlichBiomaterials) and Mucoderm® (BotissBiomaterials) matrices during the initial phase of soft tissue formation. For this purpose, samples of 5 × 5 mm were obtained, and then human fibroblasts were plated on them. After 24, 48 and 72 h, cell viability was assessed using an MTT assay, and the secretion of type I collagen, MMP-2, TIMP-1 and TIMP-2 was analyzed by ELISA immunoassay. The control group (C) consisted of cells plated on polystyrene without the matrices. The morphology of the surfaces was also examined using scanning electron microscopy (SEM), as was the average roughness (Ra) of the samples by a profilometer. Topographic analysis revealed that roughness was significantly higher on Mucoderm® than on Fibro-Gide® (p 0.05). The synthesis of type I collagen, MMP-2 and TIMP-1 were significantly higher from cells plated on Fibro-Gide® than on Mucoderm®, in all time points (p ® than on Mucoderm® (p ® induced an increase in type I collagen, MMP-2 and TIMP-1 and TIMP-2.

Effects of destruxins on free calcium and hydrogen ions in insect hemocytes

Destruxins, cyclohexadepsipeptidic mycotoxins isolated from the ento mopathogenic fungus Metarhizium anisopliae, inhibit innate insect immunity. However, their mechanism of action remains unclear. In this study, the effects ofdestruxins on changes in free calcium and hydrogen ions in the hemocytes ofExolontha serrulata, Bombyx mori and the Spodoptera litura SL1 cell line were detected using laser scanning confocal mi croscopy （LSCM）. An instant Ca2＋ influx of hemocytes induced by destruxins A and B （DA and DB） was recorded. The DA/DBdependent Ca2＋ influx was not influenced by the Ca2＋ channel inhibitors 2aminoethoxydiphenyl borane （2APB） and U73122. It also had an apparently different LSCM profile from that of the ionomycindependent Ca2＋ influx. However, the instant Ca2＋ influx was not seen in the SL1 cells; on the contrary, a slow, moderate enhancement of intracellular Ca2＋ was observed. Meanwhile, an instant intracellular free H＋ decrease aroused by DA and DB was found. DB at 20/zmol/L and DA at 690/zmol/L significantly reduced intracellular free H＋ levels. Furthermore, the vacuolar H＋ATPase （VATPase） inhibitor bafilomycin A1 had obvious effects on the decreases ofintracellular free H＋ in hemocytes. These results suggest that the mechanism of DA/DBdependent Ca2＋ influx is perhaps not related to Ca2＋ channels and ionophores; rather, the intracellular free H＋ decrease might be due to VATPase inhibition.

A key role for PTP1B in dendritic cell maturation, migration, and T cell activation

Dendritic cells(DC)are the major antigen-presenting cells bridging innate and adaptive immunity,a function they perform by converting quiescent DC to active,mature DC with the capacity to activate naı¨ve T cells.They do this by migrating from the tissues to the T cell area of the secondary lymphoid tissues.Here,wedemonstrate thatmyeloid cell-specific genetic deletion of PTP1B(LysM PTP1B)leads to defects in lipopolysaccharide-driven bone marrow-derivedDC(BMDC)activation associated with increased levels of phosphorylated Stat3.We showthatmyeloid cell-specific PTP1Bdeletion also causes decreased migratory capacity of epidermal DC,aswell as reduced CCR7 expression and chemotaxis to CCL19 by BMDC.PTP1B deficiency in BMDC also impairs their migration in vivo.Further,immature LysM PTP1B BMDC display fewer podosomes,increased levels of phosphorylated Src at tyrosine 527,and loss of Src localization to podosome puncta.In co-culture with T cells,LysM PTP1B BMDC establish fewer and shorter contacts than control BMDC.Finally,LysMPTP1BBMDCfail to present antigen to T cells as efficiently as controlBMDC.These data provide first evidence for a key regulatory role for PTP1B in mediating a central DC function of initiating adaptive immune responses in response to innate immune cell activation.

Genetics analysis of haptoglobin gene in Fujian Han nationality

To study the genetic features(characteristics)of haptoglobin gene,four different age groups of Fujian Han people were investigated.The phenotypes of the hap-toglobin of four different groups were analyzed by using polyacrylamide gel electrophoresis.The frequency of Hp^(1) in the population of Fujian Han nationality accounted for 0.340,among which children,youths,middle aged and elder groups were 0.307,0.338,0.363 and 0.383,respect-ively.The Hp^(0-0)phenotype frequency was 0.026 in which the four age groups accounted for 0.032,0.046,0.014 and 0.014,respectively.The frequency of Hp^(1)gene is rising with increasing age.The frequency of Hp^(0-0)phenotype is highest in the middle aged group and then tends to drop with increasing age.

The shrimp IKK-NF-KB signaling pathway regulates antimicrobial peptide expression and may be subverted by white spot syndrome virus to facilitate viral gene expression

The IκB kinases IKKα and IKKβ and the IKK-related kinases TANK-binding kinase 1 （TBK1） and IKKε are the master regulators of the NF-κB signaling pathway. Although this pathway has been extensively studied in mammals, less attention has been paid in crustaceans, which have significant economic value. Here, we report the cloning and functional studies of two IKK homologs, LvlKKβ and LvlKKε, from Pacific white shrimp, Litopenaeus vannamei. LvlKKβ and LvlKKα mRNAs are widely expressed in different tissues and are responsive to white spot syndrome virus （WSSV） infection. When overexpressed in Drosophila S2 cells, LvlKKβ but not LvlKKε activates the promoters of NF-κB pathway-controlled antimicrobial peptide genes （AMPs）, such as the Penaeidins （PENs）. In HEK 293T cells, both LvlKKβ and LvlKKε activate an NF-κB reporter. The silencing of LvlKKβ or LvlKKε using double-stranded RNA （dsRNA）-mediated RNA interference （RNAi） decreases the expression of L. vannamei AMPs, including PENs, lysozyme and crustins. Intriguingly, LvlKKβ- or LvlKKε-silenced L. vannameiare resistant to WSSV infection. We hypothesized that successful infection with WSSV requires the activation of the IKK-NF-κB signaling pathway to modulate viral gene expression. We constructed luciferase reporters for 147 WSSV genes. By screening, we found that the WSSV051, WSSV059, WSSV069, WSSV083, WSSV090, WSSV107, WSSV244, WSSV303, WSSV371 and WSSV445 promoters can be activated by LvlKKβ or LvlKKε in Drosophila S2 cells. Taken together, our results reveal that LvlKKβ and LvlKKε may participate in the regulation of shrimp AMPs and that WSSV may subvert the L. vannamei IKK-NF-κB signaling pathway to facilitate viral gene expression.

Molecular mechanisms underpinning sarcomas and implications for current and future therapy

Sarcomas are complex mesenchymal neoplasms with a poor prognosis.Their clinical management is highly challenging due to their heterogeneity and insensitivity to current treatments.Although there have been advances in understanding specific genomic alterations and genetic mutations driving sarcomagenesis,the underlying molecular mechanisms,which are likely to be unique for each sarcoma subtype,are not fully understood.This is in part due to a lack of consensus on the cells of origin,but there is now mounting evidence that they originate from mesenchymal stromal/stem cells(MSCs).To identify novel treatment strategies for sarcomas,research in recent years has adopted a mechanism-based search for molecular markers for targeted therapy which has included recapitulating sarcomagenesis using in vitro and in vivo MSC models.This review provides a comprehensive up to date overview of the molecular mechanisms that underpin sarcomagenesis,the contribution of MSCs to modelling sarcomagenesis in vivo,as well as novel topics such as the role of epithelial-to-mesenchymal-transition(EMT)/mesenchymal-to-epithelial-transition(MET)plasticity,exosomes,and microRNAs in sarcomagenesis.It also reviews current therapeutic options including ongoing pre-clinical and clinical studies for targeted sarcoma therapy and discusses new therapeutic avenues such as targeting recently identified molecular pathways and key transcription factors.

SCREENING FOR HUMAN CDC2H GENOMIC DNA CLONE'

The concept of the cell division cycle gene (cdc gene) originated from genetic studies on the yeast Schizosaccharomyces pombe. Among the 40—50 cdc genes known to be required specifically for cell cycle progression in the budding and fission yeasts, a most interesting cdc2 gene isolated from yeast S. pombe has attracted particular attention. It was shown