Mutual regulation of microglia and astrocytes after Gas6 inhibits spinal cord injury

Invasive inflammation and excessive scar formation are the main reasons for the difficulty in repairing nervous tissue after spinal cord injury.Microglia and astrocytes play key roles in the spinal cord injury micro-environment and share a close interaction.However,the mechanisms involved remain unclear.In this study,we found that after spinal cord injury,resting microglia(M0)were polarized into pro-inflammatory phenotypes(MG1 and MG3),while resting astrocytes were polarized into reactive and scar-forming phenotypes.The expression of growth arrest-specific 6(Gas6)and its receptor Axl were significantly down-regulated in microglia and astrocytes after spinal cord injury.In vitro experiments showed that Gas6 had negative effects on the polarization of reactive astrocytes and pro-inflammatory microglia,and even inhibited the cross-regulation between them.We further demonstrated that Gas6 can inhibit the polarization of reactive astrocytes by suppressing the activation of the Yes-associated protein signaling pathway.This,in turn,inhibited the polarization of pro-inflammatory microglia by suppressing the activation of the nuclear factor-κB/p65 and Janus kinase/signal transducer and activator of transcription signaling pathways.In vivo experiments showed that Gas6 inhibited the polarization of pro-inflammatory microglia and reactive astrocytes in the injured spinal cord,thereby promoting tissue repair and motor function recovery.Overall,Gas6 may play a role in the treatment of spinal cord injury.It can inhibit the inflammatory pathway of microglia and polarization of astrocytes,attenuate the interaction between microglia and astrocytes in the inflammatory microenvironment,and thereby alleviate local inflammation and reduce scar formation in the spinal cord.

Phylogenetic, phylogeographic and divergence time analysis of Anopheles subpictus species complex using ITS2 and COI sequences

Objective:To address the phylogenetic and phylogeographic relationship between different lineages of Anopheles(An.)subpictus species complex in most parts of the Asian continent by maximum utilization of Internal Transcriber Spacer 2(ITS2)and cytochrome C oxidase I(COI)sequences deposited at the GenBank.Methods:Seventy-five ITS2,210 COI and 26 concatenated sequences available in the NCBI database were used.Phylogenetic analysis was performed using Bayesian likelihood trees,whereas median-joining haplotype networks and time-scale divergence trees were generated for phylogeographic analysis.Genetic diversity indices and genetic differentiation were also calculated.Results:Two genetically divergent molecular forms of An.subpictus species complex corresponding to sibling species A and B are established.Species A evolved around 37-82 million years ago in Sri Lanka,India,and the Netherlands,and species B evolved around 22-79 million years ago in Sri Lanka,India,and Myanmar.Vietnam,Thailand,and Cambodia have two molecular forms:one is phylogenetically similar to species B.Other forms differ from species A and B and evolved recently in the above mentioned countries,Indonesia and the Philippines.Genetic subdivision among Sri Lanka,India,and the Netherlands is almost absent.A substantial genetic differentiation was obtained for some populations due to isolation by large geographical distances.Genetic diversity indices reveal the presence of a long-established stable mosquito population,at mutation-drift equilibrium,regardless of population fluctuations.Conclusions:An.subpictus species complex consists of more than two genetically divergent molecular forms.Species A is highly divergent from the rest.Sri Lanka and India contain only species A and B.

Genetic Polymorphism of Eighteen Lycium barbarum Resources Based on nrDNA ITS Sequence

[Objective] The study aimed to investigate the genetic polymorphism of eighteen Lycium barbarum resources via nrDNA ITS sequencing. [Method] The genomic DNAs from Lycium barbarum leaves were isolated by modified CTAB method for PCR amplification on the nrDNA ITS region using specifically synthesized primers; the amplified fragments were cloned and sequenced, then the sequencing results were clustered. [Result] nrDNA ITS sequences of the tested eighteen Lycium barbarum were firstly obtained in the present study. For all eighteen tested materials, the variation range of whole ITS region was 559-634 bp, with an average of 612 bp; alignment analyses showed that the whole length of internal transcribed spacer (ITS1+ITS2) was 480 bp, within which there are 194 variation sites (accounting for 40.4%) and 286 conserved sites (accounting for 59.6%). The cluster results showed that the eighteen tested materials could be grouped into three classes. [Conclusion] Analysis of nrDNA ITS sequence may avail to identify the Lycium barbarum germplasm resources.

ITS1 SEQUENCES OF NUCLEAR RIBOSOMAL DNA IN WILD RICES AND CULTIVATED RICES OF CHINA AND THEIR PHYLOGENETIC IMPLICATIONS

The first internal transcribed spacer(ITS1) of nuclear ribosomal DNA of three wild rice species and two subspecies of cultivated rice, which are distributed in China, was amplified using PCR technique and sequenced with automated fluorescent sequencing. The sequences of ITS1 ranged from 193 bp to 218 bp in size and G/C content varied from 69.3% to 72.7%. In pairwise comparisons among the five taxa, sequence site divergence ranged from 1.5% to 10.6%. Phylogenetic analysis of ITS1 sequences using Wagner parsimony generated a single well resolved tree, which revealed that Oryza rufipogon was much more closely related to cultivated rice species than to the other two wild species. Oryza granulata was less closely related to either cultivated rice species or the other two wild species, and might be a unique and isolated taxon in the genus Oryza. The phylogenetic relationships of the three wild rice species and two cultivated rice subspecies inferred from ITS1 sequences is highly concordant with those based on the molecular evidence from isozyme, chloroplast DNA (cpDNA), mitochondrial DNA(mtDNA) and nuclear DNA (nDNA) of the genus Oryza .

Classification and Identification of Marine Penicillium Species Based on ITS Sequences of rDNA

[ Objective ] The aim of this study is to identify 6 marine fungi species by analyzing ITS nucleotide sequence, which had been primarily identified as penicillium based on morphological characteristics. [ Method ] The ITS regions of these species were cloned by molecule biology method and phylogenetic analyzed using ClustalX1.83 software. [ Result] The ITS regions of these species were sequenced, and phylogenetic analysis between the yield sequences and the ITS sequences assessed in GenBank showed that the 6 strains all belonged to penicillium. [ Condusion] The present study suggests ITS sequence analysis could not be used as an only proof, but it is a very useful supplementary tool for the classification and identification of marine peniciUium combined with morphological characteristics.

Phylogenetic Relationships of Caragana(Fabaceae) by the Use of nrITS Sequences

[Objective] The aim was to explore the phylogenetic relationships of Cara- gana （Fabaceae） by the use of nrlTS sequences. [Method] Internal transcribed spac- er （ITS） sequences of nuclear ribosomal DNA （nrDNA） from 29 taxa of Caragana species and seven close relatives （all belong to Astralinae （Adens） Benth） were used for phylogenetic analysis. [Result] Length of the entire ITS region ranges from 611 to 614 bp in Caragana species. The aligned sequences nrlTS of Caragana are 655 bp, and 170 sites are variable, with 107 phylogenetically informative sites. The phylogenetically informative sites are 16.3% of the total aligned sequences. The ITS sequences data are useful to resolve some relationships at lower taxonomic levels within Caragana. The Caragana Fabr. is not a monophyletic group with very close connection with Calophaca tianschanica. The ITS data revealed that the species of Sect. tragacanthoides were dispersed in MP tree or ME tree. Although the morphol- ogy of C. ordosica is similar to C. tibetica, the ITS results revealed an unexpectedly close relativeship to C. roborovskyi. The ITS data also indicate C. davazamcii, C. korshinskii, C. intermedia, and C. microphylla are different species. [Conclusion] ITS sequences have an important reference value in exploring the relationships of Cara- gana.

Isolation of Rice EPSP Synthase cDNA and Its Sequence Analysis and Copy Number Determination

In order to isolate the total cDNA of rice (Oryza sativa L.) epsps gene, RT-PCR was carried out with template of rice first-strand cDNA and primers designed according to rice EPSP synthase genomic sequence obtained in previous study. A 1 585-bp cDNA fragment was amplified and cloned. The 1 585-bp cDNA contains an open reading frame (ORF) comprising of 1 533 nucleotides (nt) which encodes a 511 residue polypepetides, including 67 amino acids chloroplast transit peptide and 444 amino acids EPSP synthase mature peptide. A comparison between the EPSP synthase of different sources indicates that the mature peptide shows more than 51% identity except for the fungi EPSP synthase and the transit peptide shows considerably less sequence conservation. The copy number of rice epsps gene is estimated to be one copy per haploid rice genome using southern blot. RT-PCR indicated that rice epsps gene is expressed in rice leaves, endosperms and roots and has the highest expression level in leaves.

Sequence Analysis of ITS Region of rDNA of Alternaria Nees. from Some Areas of China

[Objective] The study aimed to identify Alternaria Nees from some areas of China at molecular level by analyzing the rDNA ITS sequence. [ Method ] The DNA sequences coding for the 5.8S rDNA and the flanking internal transcribed spacers （ ITS1 and ITS2） were amplified by PCR with universal primers ITS4 and ITS5 and subsequently sequenced for 34 Alternaria isolates from different areas of China. [Result] Sequences analysis showed that 5.8S rDNA was 159 bp and no variation in tested 34 isolates. There had variables sites in ITS. The isolates that had same sequences as A. tenuissima or A. alternata all put up eurytopicity to area and host. The variables sites of the isolates showed the diver- sity of Alternaria in the hosts of Oleaceae, Rosaceae and Solanaceae. At the same time that ITS could not clearly separated the isolates was indicated. The results indicated that the phylogenetic relationship were not closely related to the geographical origin and hosts of these isolates. [ Conclusion] The sequence analysis of ITS region could provide theory basis for the identification of Alternaria Nees..

Preliminary Study on the ITS Sequences of nrDNA from Rhodiola alsia

[Objective] The study aimed to analyze the ITS sequences of nrDNA from Rhodiola alisa and investigate the difference of evolution rate between nrDNA and trnS-trnG and rpl20-rps12 sequences of cpDNA(chloroplast DNA).[Method]Total DNA was extracted from silica-dried leaves of R.alsia by using modified CTAB method.With the extracted DNA sample as template,nrDNA ITS region was amplified,then purified and sequenced.In addition,the yielded ITS sequences were also compared with the known trnS-trnG and rpl20-rps12 sequences of cpDNA from R.alsia.[Result]The ITS sequence of nrDNA from R.alsia was 701 bp in length,of which 13 variable sites were found with a percentage of 1.85%.Of the 13 variable sites,8 were caused by point mutations,5 were the results of insertions or deletions.The(A+T)content and(G+C)content were 46.9% and 53.1%,respectively.The nucleotide diversity(π)was 0.004 27.[Conclusion]The ITS region of nrDNA from R.alsia was more conservative and evolved more slowly than the trnS-trnG and rpl20-rps12 sequences of its cpDNA.

Identification and rDNA-ITS Sequence Analysis of a New Anthracnose Pathogen of Dracaena fragrans in Yunnan Province

In this study, identification and rDNA-ITS sequence analysis of an anthracnose pathogen on Dracaena fragrans were carried out. [Method] D. fra-grans leaves with lesions were used as experimental materials to isolate anthrac-nose pathogen. Morphological observation, rDNA-ITS amplification and sequence analysis were performed to identify the pathogen strain. [Result] Caonidia of the iso-lated anthracnose pathogen were straight or curved, el iptic to crescent, with 2-5 oil droplets, 7.5-20 × 4.5-5 μm. According to molecular phylogenetic analysis, the iso-lated pathogen strain was identified as a new species, which was named Col-letotrichum dracaena-fragrantis sp. nov. [Conclusion] This study provided theoretical basis for the prevention and control of anthracnose.

Sequencing and Analysis of ITS Sequences of Bangia atropurpurea

[Object]To providing new data for phylogenesis of Bangia atropurpurea by the sequencing of internal transcribed spacer（ITS region）of rDNA.[Method]B.atropurpurea was collected from Niangziguan Spring in Shanxi,the DNA was extracted and the primers were designed for PCR amplification so as to obtain ITS gene sequence.[Result]The homology of ITS region between B.atropurpurea and Porphyra oligospermatangia was 75%,which was 79% between B.atropurpurea and P.yezoensis.[Conclusion]Compared with other genes,the ITS sequence had a greater evolution rate.The differences in classification and distribution resulted in the sequence diversity.

Molecular Cloning and Sequence Analyses of cDNAs Encoding Seven C-type Lectin-like Protein Subunits from Daboia russellii siamensis

Total mRNA was extracted from a venom gland of snake Daboia russellii siamensis following the manufacturer＇s protocol of the PolyATtract System 1000 kit purchased from Promega Biotech. cDNAs encoding C-type lectins were amplified by RT-PCR and subcloned into a pMD18-T vector. Fourteen positive clones were selected for nucleotide sequencing and seven cDNAs encoding various snake venom C-type lectin-like protein precursors, designated as DRS-L1, DRS-L2, DRS-L3, DRS-L4 DRS-L5, DRS-L6 and DRS-L7, were obtained. Amino acid sequences of these proteins were deduced and each contains a carbohydrate recognition domain. Of all the deduced protein sequences, only DRS-L1 seemed to represent a closer sequence similarity to α subunits of other known snake venom C-type lectin-like proteins using the BLAST program. Homology comparison combined with analysis of cysteine position indicate that DRS-L1 and DRS-L2 are probably the light chain LC2 and LC1 of factor X activator from Daboia russellii siamensis venom, respectively. DRS-L3 and DRS-L4 might be the β subunits of higher molecular weight C-type lectin-like proteins while DRS-L5 and DRS-L6 might be β subunits of lower molecular weight C-type lectin-like proteins. DRS-L7 might be the β subunit of a platelet membrane glycoprotein Ib-binding protein similar to echicetin.

Mitochondrial targeting sequence of magnetoreceptor MagR:More than just targeting

Iron-sulfur clusters(ISC)are essential cofactors for proteins involved in various biological processes,such as electron transport,biosynthetic reactions,DNA repair,and gene expression regulation.ISC assembly protein IscA1(or MagR)is found within the mitochondria of most eukaryotes.Magnetoreceptor(MagR)is a highly conserved A-type iron and iron-sulfur cluster-binding protein,characterized by two distinct types of iron-sulfur clusters,[2Fe-2S]and[3Fe-4S],each conferring unique magnetic properties.MagR forms a rod-like polymer structure in complex with photoreceptive cryptochrome(Cry)and serves as a putative magnetoreceptor for retrieving geomagnetic information in animal navigation.Although the N-terminal sequences of MagR vary among species,their specific function remains unknown.In the present study,we found that the N-terminal sequences of pigeon MagR,previously thought to serve as a mitochondrial targeting signal(MTS),were not cleaved following mitochondrial entry but instead modulated the efficiency with which iron-sulfur clusters and irons are bound.Moreover,the N-terminal region of MagR was required for the formation of a stable MagR/Cry complex.Thus,the N-terminal sequences in pigeon MagR fulfil more important functional roles than just mitochondrial targeting.These results further extend our understanding of the function of MagR and provide new insights into the origin of magnetoreception from an evolutionary perspective.

Molar-tooth Carbonate Sequences and Sr Isotopes in the Neoproterozoic for Stratigraphic Correlation: Research in the Jilin-Liaoning-Xuzhou-Huaiyang Area of the Sino-Korean Plate and Its Correlation with the Yangtze Plate

Based on a study of Neoproterozoic carbonates in the Jilin-Liaoning-Xuzhou-Huaiyang area, especially its cyclic sequence stratigraphy and Sr isotopes, two maximum sea flooding events (at 820 Ma and 835 Ma) have been identified. The resulting isochronous stratigraphic correlation proves that these Precambrian strata were connected between the Qingbaikou and the Nanhuan systems with a time range from 750 Ma to 850 Ma. The disappearance of microsparite carbonate and coming of a glacial stage offer important evidence for worldwide stratigraphic correlation and open a window for further correlation of the stratigraphic successions across the Sino-Korean and Yangtze Plates. A new correlation scheme is therefore provided based on our work.

Sedimentology and Sequence Stratigraphy of Marine to Lacustrine Deltaic Deposits in a Craton Basin and Their Controlling Factors: Shan 2 Member–He 8 Member(Guadalupian–Lopingian, Permian), Southeast Ordos Basin, North China

The Shan 2 Member, Shan 1 Member and He 8 Member of the Mid-Late Permian Shanxi and lower Xiashihezi formations, in the southeastern Ordos Basin, together comprise -150 m of deltaic deposits. This sequence records an overall evolution from deep marine environment to shallow lake associated with braided river, braided river delta and meandering river delta. Core description, well log interpretation, and stable isotope analysis, including carbon, oxygen and strontium, were conducted to understand the sedimentary evolution of Shan 2 to He 8 Member. The Shanxi Formation, which consists of the Shan 2 and Shan 1 members, is characterized by a tidal-influenced meandering river delta environment and a higher j13C value and S7Sr/S6Sr ratio and a lower jlSo value. The He 8 Member, the basal part of the Xiashihezi Formation, is featured by a braided river to braided river delta system and a lower j13C value, S7Sr/S6Sr ratio, and a higher jlSo value. Four third-order depositional sequences separated by five sequence boundaries are determined. Coarsening upward sequences of the Shan 2 Member-He 8 Member indicate a general regression trend, which can be correlated to global sea-level fall occurring during the Roadian-Wuchiapingian, as also evidenced by previous published zircon U-Pb results. The coal-bearing sequence （Shanxi Formation） to non-coal-bearing sequence （He 8 Member）, as well as a decrease of 87Sr/86Sr, suggest a trend from humid to arid climates. A combined effect of sea-level drop and a small uplift at the end of Shanxi Formation are proposed.

Inverse Leaf Aging Sequence (ILAS) and Its Significance of Wheat

The implication of the revelation of the inverse leaf aging sequence（ILAS） of wheat needs to be probed in theory and practice.Since 2005,the comparison experiment of the ILAS and the conventional leaf aging sequence（CLAS） has been carried out to measure the canopy temperature and some important biological parameters of wheat.In nature,there existed the phenomenon that the wheat leaves aged in an sequence opposite to the conventional sequence and some of the leaves of ILAS wheat aged sequentially differed from those of CLAS wheat,i.e.,the penultimate leaves rather than the flag leaves aged last among the leaves at different leaf positions;in correspondence with the inverse leaf aging sequence,there appeared an unconventional leaf color structure at the late fruiting stage,which had yellow upper leaves and green lower leaves,opposite to the conventional leaf color structure that had green upper leaves and yellow lower leaves;the chlorophyll concentrations,soluble protein concentrations,transpiration rates,and net photosynthetic rates of the penultimate leaves of ILAS wheat unconventionally surpassed those of their flag leaves as their growth moved forward from one stage to another stage,and the ILAS wheat characteristically presented cold canopy temperature,i.e.,a cold tail canopy temperature at the late fruiting stage,or a cold canopy temperature at the whole fruiting stage;because ILAS wheat was unique in physiological process,its kernel weights were obviously higher than those of CLAS wheat,which closely related to the ＂relay＂ kernel-filling mode,which was different from the kernel-filling mode of CLAS wheat under which the flag leaves act as the main nutrient supply source at the whole fruiting stage.This study provided a new idea and approach for the theoretical exploration on wheat fruiting and aging,wheat yield further improvement,and cold type wheat and cold tail wheat breeding.

Effect of cobalt and its adding sequence on the catalytic performance of MoO_3/Al_2O_3 toward sulfur-resistant methanation

The effect of promoter cobalt and the sequences of adding cobalt and molybdenum precursors on the performance of sulfur-resistant methanation were investigated. All these samples were prepared by impregnation method and characterized by N2-adsorption, X-ray diffraction(XRD), temperature-programmed reduction(TPR) and laser Raman spectroscopy(LRS). The conversions of CO for Mo-Co/Al, Co-Mo/Al and CoMo/Al catalysts were 59.7%, 54.3% and 53.9%, respectively. Among these catalysts, the Mo-Co/Al catalyst prepared stepwisely by impregnating Mo precursor firstly showed the best catalytic performance. Meanwhile, the conversions of CO were 48.9% for Mo/Al catalyst and 10.5% for Co/Al catalyst. The addition of cobalt species could improve the catalytic activity of Mo/Al catalyst. The N2-adsorption results showed that Co-Mo/Al catalyst had the smallest specific surface area among these catalysts. CoMoO4species in CoMo/Al catalyst were detected with XRD, TPR and LRS. Moreover, crystal MoS2which was reported to be less active than amorphous MoS2was found in both Co-Mo/Al and CoMo/Al catalysts. Mo-Co/Al catalyst showed the best catalytic performance as it had an appropriate surface structure, i.e., no crystal MoS2and very little CoMoO4species.

Sedimentary microtopography in sequence stratigraphic framework of Upper Ordovician and its control over penecontemporaneous karstification,No.I slope break,Tazhong,Tarim block

In order to reveal the relationship between the penecontemporaneous karstification and sedimentary microtopography in sequence stratigraphy,the sequence stratigraphic framework of Lianglitage formation in Upper Ordovician is studied according to the well drilling,logging,geophysical data,detailed observations of core and the paleontology.The Lianglitage formation belongs to the sequence Ⅳ of Ordovician.The second member of Lianglitage formation is prograde sedimentation in highstand systems tract,and is favorable for developing reef flat.The development scale and thickness of reef flat are controlled by the variation of secondary sea level.The types and characteristics of karst in the highstand systems tract show that the late highstand systems tract is dissolved and cemented by the meteoric fresh water and mixed water.Penecontemporaneous karstification is developed at the top of parasequence and high place of geomorphology.Atmospheric diagenetic lens is formed.The developing regulations and controlling factors of penecontemporaneous karstification can provide new clues to the prediction and exploration of favorable reservoir in this area.

Architecture of volcanic sequence and its structural control of Yingcheng Formation in Songliao Basin

Yingcheng Formation is a set of volcanic strata composed of lava rocks,volcanic clastic rocks and sedimentary rocks,filled in some fault depressions in Songliao Basin,early Cretaceous.The study about litho-facies succession of Yingcheng Formation in the outcropped area of the southeast margin and in Xujiaweizi fault depression and its distribution based on analysis of seismic data,shows that the sequence of volcanic strata is quite different from the clastic sedimentary sequence.To study the architecture of volcanic sequence and its structural control of Yingcheng Formation in Songliao Basin,in this work,dividing of the volcanic sequence and dating of the sequence boundaries were finished firstly,then displacement and displacement rate of faults were calculated.The results show that,sample ages of top of the first member,the seconde member,and the third member are 127 Ma,115 Ma,110.7 Ma,respectively and sample age of the bottom of the third member is 114.7 Ma.The maximum displacement and displacement rate of the fault 1 are 3 km and 300 m/Ma,respectively,and those of the fault 2 are 3 km and 1000 m/Ma.Studies suggest that,the cooling unit of lava rock or pyroclastic rock is a basic genetic stratigraphic unit in volcanic sequence stratigraphy.Cooling units can construct a parasequence reflecting a volcanic eruption stage.A sequence was superimposed by some parasequences,responding to a volcanic active cycle.There are three types of volcanic sequences in Yingcheng Formation: type of explosion,type of effusion and type of mixed explosion-effusion.The surface of the volcanic sequence,an unconformity surface widely spread and traced in seismic profiles,is a base for analysis of volcanic sequence.The development of volcanic sequence was controlled by faulting,and the curves of fault displacement(rate)can reflect this control.The preservation of volcanic sequence was controlled by the type of volcanic structure and the regional subsidence,also different from that of the sedimentary.The type of volcanic structure of Xujiaweizi was a volcanic depression during the forming of Yingcheng Formation,and the breakdowns of volcanoes and structural subsidence were key factors in the volcanic sequences preservation.

A preliminary analysis of phylogenetic relationships of Arundinaria and related genera based on nucleotide sequences of nrDNA （ITS region） and cpDNA （trnL-F intergenic spacer）

Phylogenetic relationships of Arundinaria and related genera (Pleioblastus, Pseudosasa, Oligostachyum, Bashania, Clavinodum, etc.) were assessed by analyzing the sequences of the nrDNA internal transcribed spacer (ITS) and the cpDNA trnL-F intergenic spacer (IGS). Comparison with trnL-F IGS sequence, the ITS region provided the higher number of parsimony informative characters, and the interspecific variation of the ITS sequence was higher than that of the trnL-F IGS sequence.The tree obtained by combining both sets of data showed that the species sampled in Arundinaria and the related genera were monophyletic and divided into two clades. The relationships and positioning of all the taxa surveryed (including A. oleosa, A. hsienchuensis, A. chino, A. amara, A. yixingensis, A. amabilis, A. fortunei, A. pygmaea, A. gramineus, A. fargesii, A. faberi, A. hupehense, Pseudosasa japonica cv. Tsutsumiana, P. japonica and Brachystachyum densiflorum) were also discussed. The results from the sequences were broadly consistent with morphological characters, appearing all these taxa sampled belong to the genus of Arundinaria. The topologies of the trees generated from individual data and the combined data were similar.