Correlation of free radical level and apoptosis after intracerebral hemorrhage in rats

Objective To investigate the correlation of perihematomal free radical level and neuronal apoptosis following the intracerebral hemorrhage （ICH）. Methods Animals were randomly divided into 4 groups： sham operation group, model group, 1 mg/kg edaravone group, and 3 mg/kg edaravone group. Each group was then divided into seven subgroups, in which the rats were correspondingly killed at 6 h, 12 h, 24 h, 48 h, 72 h, 7 d or 14 d （n = 1 in each subgroup of the sham group, and n = 6 in each subgroup of the other 3 groups）. By Horseley-Clarke technique, autoblood （80 μL） were administered into the left caudate putamen of SD rats in a double administration-withdrawal way. Rats in the sham group were needled in but not administered with autoblood. The ICH model was then evaluated by Bederson＇s scale. Around the hematoma, the levels of malonaldehyde （MDA） and hydroxyl radical were tested by spectrophotometer, and the process of apoptosis was tested by terminal deoxynucleofidyltransferase-mediated dUTP-biotin nick end labeling （TUNEL） method. Results （1） ICH significantly increased the levels of MDA and hydroxyl radicals. Significant differences in MDA and hydroxyl radical contents were observed among the four groups. （2） In the sham group, a small number of TUNEL-positive cells were found. In the other three groups, the TUNEL-positive cells were observed at 6 h, increased significantly at 24 h, and reached peak level at 3 d, then fell profoundly at 7 d, but remained detectable at 14 d. （3） The positive correlation existed between apoptosis and free radical level （r = 0.2003）, and existed between apoptosis and MDA content （r = 0.6563） in the brain. Conclusion Post-hemorrhagic apoptosis was related to the production of free radicals, indicating that the elevated free radicals following the ICH could induce neuron and glial cell apoptosis.

Study on Establishment of cpSSR Marker Technology and Optimization of Its Reaction System in Jatropha curcas Linn

[ Objective] The aim of this study was to establish the optimum cpSSR-PCR system for Jatropha curcas Linn. [ Method] cpSSR-PCR amplification system for Jatropha curcas Linn influenced by five factors including Taq DNA polymerase, Mg^2＋ , DNA template, dNTP and primer were optimized from several levels. [ Result] The optimum concentration of 20 μl reaction system was 10 × Buffer, 2.00 mmol/L Mg^2＋ , 2 U/μl Taq DNA polymerase, 0.2 mmol/L dNTP, 0.2 μmol/L primer and 35 ng/μl DNA template. [ Conclusion] The optimum annealing temperature for cpSSR-PCR reaction system is 52 ℃, and the cpSSR reaction system is steady and reproducible.

Mechanized Seed Production of Xinhunyou No.6 through Mixed Seeding of Parents' Seeds

Hybrid rice Xinhunyou No. 6 could be cultivated by mixing male and fe- male parents and performing seedling raising followed by transplanting, mechanical planting or mechanical direct seeding according to the production methods of com- mercial rice. During flowering stage, leaf cutting, application of ＂920＂ and pollination were conducted; after pollination, bentazon with a certain concentration was sprayed to kill the male parent; and the hybrid rice was harvested mechanically. Before storage, color separation was performed to remove little remaining male parent, thereby achieving whole-process mechanization of hybrid rice seed production. This study introduced mechanized seed production of Xinhunyou No. 6 through mixed- seeding from the aspects including mechanical direct seeding, seeding raising fol- lowed by transplanting and mechanical planting.

Discussion on AFLP Molecular Markers in Piper methysticum and Pepper

The aim of the research was to discuss the genetic relationships between Piper methysticum, Pepper and other wild species in Pepper genus. DNA was extracted from leaves which belonged to 28 germplasms including 6 materials of P. methysticum, 21 maerials of cultivated and wild Pepper, 1 material of Peperomia pellucida belonged to different genus. Premiers with good band-type and high polymorphism and resolution were selected from 64 pairs of primers for AFLP amplification and the clustering analysis was conducted with MVSP3.13f software. 191 bands were amplified by 4 pairs of premiers, 189 of which had polymorphism, being 98.6%. 28 germplasms were classified into 6 different groups at the genetic similarity coefficient of 0.52 by silver staining AFLP, in which 6 materials of Piper methysticum were clustered into a single group, indicating that P. methysticum belonged to Pepper family of Pepper genus but were distantly related to the others. The research provided the basis for selecting rootstocks for P. methysticum graft, molecular identification of P. methysticum and the fingerprint construction of P. methysticum.

银屑病患者皮损处CD4^+ CD25^+ Foxp3^+调控T细胞的识别

Background: Depletion of CD4+CD25+Foxp3+naturally occurring regulatory T cells (Treg) induces autoimmune phenomena. These cells have not yet been fully characterized in the skin of psoriatic patients. Objectives: To prove that the Zenon immunofluorescent labeling technique is suitable for the demonstration of co-localization of T-cell markers and in particular to show the distribution of Treg in psoriatic skin. Methods: In biopsies derived from normal and psoriatic skin, CD4+CD25+, CD4+CD45RO+, CD8+CD25+, CD8+CD45RO+and CD4+CD25+Foxp3+cells in the dermis and in the epidermis were immunophenotyped, using a quantitative immunofluorescent labeling technique (Zenon), analyzed and compared using image analysis. Results: The immuno-fluorescent labeling technique was shown to be an easy and reliable tool to demonstrate co-localization of T-cell markers.In psoriasis, all pathogenic T-cell subsets (CD4+CD25+,CD4+CD45RO+, CD8+CD25+and CD8+CD45RO+cells) were significantly increased in the dermis and in the epidermis, as compared to normal skin (all p < 0.05). Using this labeling technique we were able to reveal CD4+CD25+Foxp3+Treg in psoriatic dermis, but not in the dermis of normal skin (p < 0.0001). Conclusions: The Zenon immunofluorescence technique in combination with image analysis is suitable for the demonstration of co-localization of T-cell markers in tissue. Increased numbers of pathogenic T cells (CD4+CD25+,CD4+CD45RO+, CD8+CD25+and CD8+CD45RO+) were shown in the dermis and epidermis, whereas CD4+CD25+Foxp3+Treg were identified in psoriatic skin with a predilection for the upper dermis.

Use of tetracycline hydrochloride and alizarin complexone for immersion marking black rockfish Sebastes schlegelii

We tested the utility of chemical marking techniques in the juvenile black rockfish Sebastes schlegelii. Juveniles （30-40 mm total length） were immersed in a range of tetracycline hydrochloride （TC） solutions at concentrations ranging from 300 to 500 mg/L, and alizarin complexone （ALC） solutions at concentrations ranging from 200 to 400 mg/L in filtered sea water （salinity of 30） for 24 h, respectively. Otoliths （sagittae, asteriscus）, scales, fin rays （dorsal, pectoral, ventral, anal, and caudal fin rays）, and fin spines （dorsal, ventral, and anal fin spines） were sampled and used to detect fluorescent marks after a 60-day growth experiment. With the exception of 300 mg/L TC, acceptable marks were produced in the otoliths and fin spines by all concentrations of TC and ALC. In particular, we observed clearly visible marks in the sagittae, asteriscus, and fin spines under normal light at concentrations of200~00 mg/L, 250-400 mg/L, and 250-400 mg/L ALC, respectively. Scales and fin rays had acceptable marks at much higher concentrations （_〉350 mg/L TC, 〉250 mg/L ALC for scales and _〉350 mg/L TC, 〉300 mg/L ALC for fin rays）. The best mark quality （i.e., acceptable marks were observed in all sampled structures after immersion marking） were obtained following immersion in TC at between 350-500 rag/L, and ALC between 300-400 mg/L. In addition, there was no significant difference in survival and growth of TC and ALC marked fish compared to their controls up to 60 days post-marking （P〉0.05）.

Progress on Mapping of Quantitative Trait Loci in Crops

Most important agricultural traits of crops are controlled by many genes. These traits have complicated genetic basis and are difficult for genetic analysis. Due to application of molecular marker techniques in the last two decades, genetic and molecular dissection of quantitative traits has become possible. In this paper, recent progress on mapping of quantitative trait loci in crops was reviewed.

Relaxation labeling for non-rigid point matching under neighbor preserving

Non-rigid point matching has received more and more attention.Recently,many works have been developed to discover global relationships in the point set which is treated as an instance of a joint distribution.However,the local relationship among neighboring points is more effective under non-rigid transformations.Thus,a new algorithm taking advantage of shape context and relaxation labeling technique,called SC-RL,is proposed for non-rigid point matching.It is a strategy that joints estimation for correspondences as well as the transformation.In this work,correspondence assignment is treated as a soft-assign process in which the matching probability is updated by relaxation labeling technique with a newly defined compatibility coefficient.The compatibility coefficient is one or zero depending on whether neighboring points preserving their relative position in a local coordinate system.The comparative analysis has been performed against four state-of-the-art algorithms including SC,ICP,TPS-RPM and RPM-LNS,and the results denote that SC-RL performs better in the presence of deformations,outliers and noise.

Characterization of enhancer trap and gene trap harboring Ac／Ds transposon in transgenic rice

Insertion mutagenesis has become one of the most popular methods for gene functions analysis.Here we report a two-element Ac/Ds transposon system containing enhancer trap and gene trap for gene tagging in rice.The excision of Ds element was examined by PCR amplification.The excision frequency of Ds element varied from 0% to 40% among 20 F2 populations derived from 11 different Ds parents.Southern blot analysis revealed that more than 70% of excised Ds elements reinserted into rice genome and above 70% of the reinserted Ds elements were located at different positions of the chromosome in rice.The result of histochemical GUS analysis indicated that 28% of enhancer trap and 22% of gene trap tagging plants displayed GUS activity in leaves, roots,flowers or seeds.The GUS positive lines will be useful for identifying gene function in rice.

Intestinal microflora molecular markers of spleen-deficient rats and evaluation of traditional Chinese drugs

AIM:To find a rapid and efficient analysis method of gastrointestinal microflora in Pi-deficient(spleen-deficient) rats and to evaluate traditional Chinese drugs.METHODS:Enterobacterial repetitive intergenic consensus-PCR(ERIC-PCR) based assay was performed to examine changes of intestinal microflora in two Pi-deficienct animal models and to evaluate the efficacy of four traditional Chinese drugs as well as a probiotic recipe and another therapy in Pi-deficient rats.RESULTS:A molecular marker was identified for Pi-deficiency in rats.The pharmacodynamic evaluation system,including identified molecular markers(net integral area and abundance of DNA bands),Shannon's index for diversity of intestinal microflora,and Sorenson's pairwise similarity coefficient,was established.The four major clinical recipes of traditional Chinese drugs for Pi-deficiency in rats,especially at their medium dose(equivalence to the clinical dose),produced more pronounced recovery activities in Pi-deficient rats,while higher doses of these recipes did not show a better therapeutic effect but some toxic effects such as perturbation deterioration of intestinal microflora.CONCLUSION:Both fingerprint analysis and identified marker can show Pi-deficiency in rats and its difference after treatment.The identified molecular marker may be applied in screening for the active compounds both in relative traditional Chinese drugs and in pharmacodynamic study of Pi-deficiency in rats.

LTAG Technology Passed Appraisal

Recently the LTAG technology combining selective hydro-saturation of LCO with selective catalytic cracking technology for producing high-octane gasoline or light aromatic hydrocarbons has passed technical appraisal.

Development of a SCAR marker for male gametophyte of Gracilariopsis lemaneiformis based on AFLP technique

The red alga Gracilariopsis lemaneiformis(Bory) is an economically valuable macroalgae. As a means to identify the sex of immature Gracilariopsis lemaneiformis, the amplifi ed fragment length polymorphism(AFLP) technique was used to search for possible sex- or phase-related markers in male gametophytes, female gametophytes, and tetrasporophytes, respectively. Seven AFLP selective amplifi cation primers were used in this study. The primer combination E-TG/M-CCA detected a specifi c band linked to male gametophytes. The DNA fragment was recovered and a 402-bp fragment was sequenced. However, no DNA sequence match was found in public databases. Sequence characterized amplifi ed region(SCAR) primers were designed from the sequence to test the repeatability of the relationship to the sex, using 69 male gametophytes, 139 female gametophytes, and 47 tetrasporophytes. The test results demonstrate a good linkage and repeatability of the SCAR marker to sex. The SCAR primers developed in this study could reduce the time required for sex identifi cation of Gracilariopsis lemaneiformis by four to six months. This can reduce both the time investment and number of specimens required in breeding experiments.

High-efficiency 18S microalgae barcoding by coalescent,distance and character-based approaches: a test in Chlorella and Scenedesmus

The relatively conserved 18S is often used in the phylogenetic analysis of microalgae. However, whether it can really help in barcoding microalgae needs to be evaluated. In this study the multiple approaches of coalescent, distance and character-based barcoding are first employed in C hlorella and Scenedesmus to test the efficiency of 18S sequences for barcoding green microalgae. We show that most Chlorella and Scenedesmus species, including the cryptic species, can be distinguished by 18S sequences with all coalescent General Mixed Yule-coalescent(GMYC), poisson tree process(PTP), and P ID, distance(ABGD) and character-based approaches. Both GMYC and PTP analyses produce more genetic groups. The P ID and ABGD analyses only cluster some species. All species(apart from a few of lineages) can be separated in character-based barcoding analysis with more than three character attributes. In comparison with previous barcoding results with r bcL, tufA, ITS and 16 S, 18S produces good resolution in identifying Chlorella and Scenedesmus. Our results reveal that 18S is highly efficient in identifying taxa of green microalgae at species level, based on a combination of multiple barcoding approaches. Combining 18S with other gene markers may be useful in barcoding microalgae.

Paternity Testing of Selected Abaca （Musa textilis L. Nee） Hybrids Using Morphometric Markers

Investigation into the paternity of four abaca （Musa textilis L, Nee） hybrids was done to ascertain the mode of transmission of selected morpho-agronomic traits and to detect possible heterosis. In situ morphological characterization was undertaken using twenty five qualitative and six quantitative characters. Results revealed that a great majority of the qualitative traits were shared by both parents and their hybrids. For the rest, the qualitative traits were inherited from one or the other parent though some variant phenotypes （i.e. chimerism） were also noted in the hybrids. Cases ofheterosis were also observed and this could be exploited to increase fiber yield in the hybrids. Though inconclusive due to factors such as the heterogenous nature of abaca plants in the field and the susceptibility of morphological traits to environmental fluctuations, this study has provided baseline information on abaca hybridity that can be verified using more robust technologies as molecular markers.

Clones identification of Sequoia sempervirens (D. Don) Endl. in Chile by using PCR-RAPDs technique

A protocol of polymerase chain reaction-random amplified polymorphic DNAs (PCR-RAPDs) was established to analyse the gene diversity and genotype identification for clones of Sequoia sempervirens (D. Don) Endl. in Chile. Ten (out of 34) clones from introduction trial located in Voipir-Villarrica, Chile, were studied. The PCR-RAPDs technique and a modified hexadecyltrimethylammonium bromide (CTAB) protocol were used for genomic DNA extraction. The PCR tests were carried out employing 10-mer random primers. The amplification products were detected by electrophoresis in agarose gels. Forty nine polymorphic bands were obtained with the selected primers (BG04, BF07, BF12, BF13, and BF14) and were ordered according to their molecular size. The genetic similarity between samples was calculated by the Jaccard index and a dendrogram was con- structed using a cluster analysis of unweighted pair group method using arithmetic averages (UPGMA). Of the primers tested, 5 (out of 60) RAPD primers were selected for their reproducibility and high polymorphism. A total of 49 polymorphic RAPD bands were detected out of 252 bands. The genetic similarity analysis demonstrates an extensive genetic variability between the tested clones and the dendrogram depicts the genetic relationships among the clones, suggesting a geographic relationship. The results indicate that the RAPD markers permitted the identification of the assayed clones, although they are derived from the same geo- graphic origin.

^(18)F-Labeling techniques for positron emission tomography

Much research effort has been made to understand various biological processes at levels of molecules using molecular imaging techniques. Because of great sensitivity, high resolution, and rapid detection, positron emission tomography （PET） imaging is becoming one of the most used imaging techniques for medical diagnose and pre-clinical studies. Here we provide a review on molecular imaging and PET imaging. An introduction is also provided on ~SF-fluorine labeling techniques for the preparation of PET imaging probes. A summary and comparison of currently available ~SF-fluorine labeling methods is provided. The per- spectives for 18F-fluorine labeling techniques are also given.

DNA-associated click chemistry

This review highlights the most recent advances in click chemistry associated with DNA.Cu[I]-catalyzed azides-alkynes Huisgen cycloadditions(CuAAC)and a strain-promoted alkyne-azide cycloaddition(SPAAC)are two popular click reactions that have great impact in DNA science.The simplicity,versatility,orthogonality,and high efficiency of click reaction along with a stable triazole product have been instrumental for the successful application of this reaction in the field of nucleic acid chemistry.CuAAC and SPAAC reactions have been widely used for DNA modification,including DNA labeling,metallization,conjugation,cross-linking,and ligation.Modified oligodeoxynucleotides obtained from click reaction have been extensively applied in the fields of drug discovery,nanotechnology,bio-conjugation,and material sciences,among others.The most recent advances in the synthesis and applications of clickable DNAs are discussed in detail in this article.

A code-based approach for labeling in complex irregular regions

Labeling information in a complex irregular region is a useful procedure occurring frequently in sheet metal and the furniture industry which will be beneficial in parts management.A fast code-based labeler(FCBL) is proposed to accomplish this objective in this paper.The region is first discretized,and then encoded by the Freeman encoding technique for providing the 2D regional information by 1D codes with redundancies omitted.We enhance the encoding scheme to make it more suitable for our complex problem.Based on the codes,searching algorithms are designed and can be extended with customized constraints.In addition,by introducing a smart optimal direction estimation,the labeling speed and accuracy of FCBL are significantly improved.Experiments with a large range of real data gained from industrial factories demonstrate the stability and millisecond-level speed of FCBL.The proposed method has been integrated into a shipbuilding CAD system,and plays a very important role in ship parts labeling process.