Objective To summarize the experience in the diagnosis and treatment of symptomatic splanchnic artery dissection. Methods A total of 21 patients with symptomatic splanchnic artery dissection, who were admitted to the Affiliated First Hospital of China Medical University during the period from June 2006 to March 2014, were included in this study. Combined with the literature, the clinical data, including the diagnosis and treatment, were analyzed. Results Contrast-enhanced abdominal CT and CT angiography revealed superior mesenteric artery dissection in 15 cases, celiac artery dissection in 5 cases and splenic artery dissection in one case. Conservative therapy was employed in 5 patients; among them one was complicated with hepatic artery thrombosis. Of the 16 patients who received endovascular stent placement, additional intestinal resection was performed in 2 and transcatheter thrombolysis treatment in other 2. No procedure-related severe complications occurred in perioperative period. All the patients were followed up for 2-74 months (mean of 19.1 months). In patients who received endovascular stent placement, the abdominal pain and the bloody stool were relieved or disappeared, and no abdominal pain recurred. CT angiography showed that in-stent blood flow was fluent, the stent was in good position, and neither stenosis nor thrombosis was observed. One patient with superior mesenteric artery dissection died of stroke three months after the treatment. Conclusion It is very important to make early diagnosis and to adopt early treatment for symptomatic splanchnic artery dissection. CT angiography can confirm the diagnosis in most cases, but attention should be paid to some atypical manifestations. For the treatment of splanchnic artery dissection, endovascular stent placement is mini-invasive, safe and reliable.

Objective To evaluate the technique and result of endovascular treatment for right subclavian artery stenosis or occlusion.Methods Seventeen patients [13 males,4 females ; (56 ± 11)years old] with right subclavian artery stenosis or occlusion were treated with endovascular surgery which included recanalization,balloon angioplasty and stenting via femoral or brachial artery route.Cerebral protection devices were used in 6 cases to avoid cerebral embolism.Results Sixteen of the seventeen patients acquired successful recanalization in 8 cases with subclavian artery stenosis (100％ technical success rate) and in other 8 cases with subclavian artery occlusion (88.9％ technical success rate).Five cases were treated with balloon angioplasty,and 11 cases were treated with balloon angioplasty combined with stenting.Good patency was seen in the 16 cases immediately after the procedure.The cerebral protection devices prevented all the cases from cerebral embolism and were retrieved suceessfully.Sixteen cases were followed up from 1 to 66 months [mean (24 ± 18) months].Restenosis was found in one case 10 months later and was successfully treated with re-PTA.One case with aortoarteritis died of cerebral infarction 18 months later.No symptom recurrence was found in other cases and ultrasound or CTA of followup showed excellent patency.Conclusions Balloon angioplasty and stenting are safe and effective for the treatment of right subclavian artery occlusion.

Objective To study the relationship between erythromycin sensitivity and drug-resistance gene in Mycoplasma pneu-moniae(Mp).Methods In 46 erythromycin-resistant MP clinical isolates,domain Ⅴ of 23S rRNA was amplified by polymerase chain reaction(PCR),followed by direct automatic sequencing method.The DNA sequences were compared to find molecular mecha-nisms of drug resistance.Results Among the 46 erythromycin-resistant Mp clinical isolates,44(95.65%)harbored an A-to-G tran-sition mutation at position 2063 in the 23S rRNA gene and 2(4.35%)harbored an A-to-G transition mutation at position 2064,but no A-to-C transition mutation at position 2063 and C-to-G/A transition mutation at position 2617 were detected.Conclusion E-rythromycin-resistant of Mp clinical isolates were closely related to A-to-G transition mutation at position 2063/2064 in domain Ⅴof 23S rRNA genes and the most important was the A2063G transition mutation.Rapid and accurate identification of the genetic mutations in domain Ⅴ of 23S rRNA may be help to diagnose the infection of Mycoplasma pneumoniae,provide the drug-resistant information,and instruct the application of antibiotics reasonably and effectively.

AIM To construct a non-normalized cDNA library from Agkistrodon acutus venom gland as an imtial step to develop new and more effective venom by genetic engineering technique for screening and expressing target genes. METHODS The total RNA was extracted from fresh venom gland using Trizol. mRNA was reversely transcripted to cDNA using superscriptⅡ reverse transcriptase. Second-strand synthesis was performed using DNA polymeraseⅠ. After adding EcoRⅠ adaptor, phosphorylating the end and digesting with XhoⅠ, the cDNA was collected in five fractions (<0.25 kb, 0.25-0.5 kb, 0.5-1 kb, 1-2 kb and >2 kb) using the QIAquick Gel Extraction kit and ligated to pBluescriptⅡ vectors. The five libraries obtained were plated by infecting E.coli DH10B, constructing a cDNA library of Agkistrodon acutus venom gland. Sequencing clones at random, 8696 high quality 5′ end expressed sequenced tags (ESTs) were obtained and analyzed. The initial sequences were assembled into 2855 clusters. Among which, one of the clusters (Agkihagin) consisting of 74 ESTs was identified as a novel metalloprtoteinase based on RT-PCR and sequence analysis. RESULTSThe titers of library were 2.048×106. The novel metalloproteinase belonged to PⅢ type metalloproteinase. Its open reading frame was composed of 1827 nucleotides and coded a pre-zymogen of 608 amino acid with zinc-binding domain for metalloproteinase and Asp-Glu-Cys-Asp(DECD) domain for disintegrin. CONCLUSION The capacity of cDNA library of venom gland is above the general level of cDNA library. It would be a helpful platform to construct a catalog for transcripts in the venom gland of the Agkistrodon acutus. The sequence analysis indicates that the deduced amino acid sequence of the identified gene for metalloproteinase share the highest 87% identity with the metalloproteinase genes of other snakes in the GenBank. It lays a good foundation for the study of structure-function relationships of snake venom metalloproteinases.

With the development of genomics and bioinformatics, especially the extensive applications of high-throughput sequencing technology, more transcriptional units with little or no protein-coding potential have been discovered. Such RNA molecules are called non-protein-coding RNAs (npcRNAs or ncRNAs). Among them, long npcRNAs or ncRNAs (lnpcRNAs or lncRNAs) represent diverse classes of transcripts longer than 200 nucleotides. In recent years, the lncRNAs have been considered as important regulators in many essential biological processes. In plants, although a large number of lncRNA transcripts have been predicted and identified in few species, our current knowledge of their biological functions is still limited. Here, we have summarized recent studies on their identification, characteristics, classification, bioinformatics, resources, and current exploration of their biological functions in plants.

Plant non-specific lipid transfer proteins(nsLtps) have been reported to be involved in plant defense activity against bacterial and fungal pathogens.In this study,we identified 135 (122 putative and 13 previously identified) Solanaceae nsLtps,which are clustered into 8 different groups.By comparing with Boutrot's nsLtp classification,we classified these eight groups into five types (Ⅰ,Ⅱ,Ⅳ,Ⅸ and Ⅹ).We compared Solanaceae nsLtps with Arabidopsis and Gramineae nsLtps and found that (1) Types Ⅰ,Ⅱ and Ⅳ are shared by Solanaceae,Gramineae and Arabidopsis;(2) Types Ⅲ,Ⅴ,Ⅵ and Ⅷ are shared by Gramineae and Arabidopsis but not detected in Solanaceae so far;(3) Type Ⅶ is only found in Gramineae whereas type Ⅸ is present only in Arabidopsis and Solanaceae;(4) Type X is a new type that accounts for 52.59% Solanaceae nsLtps in our data,and has not been reported in any other plant so far.We further built and compared the three-dimensional structures of the eight groups,and found that the major functional diversification within the nsLtp family could be predated to the monocot/dicot divergence,and many gene duplications and sequence variations had happened in the nsLtp family after the monocot/dicot divergence,especially in Solanaceae.

Objective To construct the UAS-Hey transgenic fly strains to provide a tool for researching the function of Hey gene in the fly development.Methods The Hey gene coding sequence was amplified by reverse transcription PCR and subcloned into pUAST expression vector.The pUAS-Hey recombinant plasmid was constructed and microinjected into the embryos of wild type flies.The UAS-Hey transgenic flies with red eyes were screened out with mini-white marker,then the balancing and mapping of these transgenic strains were performed.The identification was performed by using the PCR amplification method.Results The pUAS-Hey recombinant plasmid was successfully constructed,and seven independent transgenic strains were obtained by microinjection and transgenic fly screening and balance.PCR analysis confirmed that the P[mini-white,UAS-Hey] was integrated into genomes of transgenic strains and was in expressible region.Conclusion The UAS-Hey transgenic flies are successfully constructed,which lays the foundation for further studies of the function and regulation of Hey gene with GAL4/UAS system.

Our recent investigation in the protist Trichomonas vaginalis suggested a DNA sequence periodicity with a unit length of 120.9 nt, which represents a sequence signature for nucleosome positioning. We now extended our observation in higher eukaryotes and identified a similar periodicity of 175 nt in length in Caenorhabditis elegans. In the process of defining the sequence compositional characteristics, we found that the 10.5-nt periodicity, the sequence signature of DNA double helix, may not be sufficient for cross-nucleosome positioning but provides essential guiding rails to facilitate positioning. We further dissected nucleosome-protected sequences and identified a strong positive purine (AG) gradient from the 5'-end to the 3'-end, and also learnt that the nucleosome-enriched regions are GC-rich as compared to the nucleosome-free sequences as purine content is positively correlated with GC content. Sequence characterization allowed us to develop a hidden Markov model (HMM) algorithm for decoding nucleosome positioning computationally, and based on a set of training data from the fifth chromosome of C. Elegans, our algorithm predicted 60%-70% of the well-positioned nucleosomes, which is 15%-20% higher than random positioning. We concluded that nucleosomes are not randomly positioned on DNA sequences and yet bind to different genome regions with variable stability, well-positioned nucleosomes leave sequence signatures on DNA, and statistical positioning of nucleosomes across genome can be decoded computationally based on these sequence signatures.

The emergence of next-generation sequencing (NGS) technologies has significantly improved sequencing throughput and reduced costs.However,the short read length,duplicate reads and massive volume of data make the data processing much more difficult and complicated than the first-generation sequencing technology.Although there are some software packages developed to assess the data quality,those packages either are not easily available to users or require bioinformatics skills and computer resources.Moreover,almost all the quality assessment software currently available didn't taken into account the sequencing errors when dealing with the duplicate assessment in NGS data.Here,we present a new user-friendly quality assessment software package called BIGpre,which works for both Illumina and 454 platforms.BIGpre contains all the functions of other quality assessment software,such as the correlation between forward and reverse reads,read GC-content distribution,and base Ns quality.More importantly,BIGpre incorporates associated programs to detect and remove duplicate reads after taking sequencing errors into account and trimming low quality reads from raw data as well.BIGpre is primarily written in Peal and integrates graphical capability from the statstics package R.This package produces both tabular and graphical summaries of data quality for sequencing datasets from Illumina and 454 platforms.Processing hundreds of millions reads within minutes,this package provides immediate diagnostic information for user to manipulate sequencing data for downstream analyses. BIGpre is freely available at http://bigpre.sourceforge.net/.

Histone H3 lysine 4 trimethylation (H3K4me3) is well known to occur in the promoter region of genes for transcription activation.However,when investigating the H3K4me3 profiles in the mouse cerebrum and testis,we discovered that H3K4me3 also has a significant enrichment at the 3' end of actively transcribed (sense) genes,named as 3′-H3K4me3.3′-H3K4me3 is associated with ～15％ of protein-coding genes in both tissues.In addition,we examined the transcriptional initiation signals including RNA polymerase II (RNAPII)binding sites and 5′-CAGE-tag that marks transcriptional start sites.Interestingly,we found that 3′-H3K4me3 is associated with the initiation of antisense transcription.Furthermore,3′-H3K4me3 modification levels correlate positively with the antisense expression levels of the associated sense genes,implying that 3′-H3K4me3 is involved in the activation of antisense transcription.Taken together,our findings suggest that H3K4me3 may be involved in the regulation of antisense transcription that initiates from the 3′ end of sense genes.In addition,a positive correlation was also observed between the expression of antisense and the associated sense genes with 3'-H3K4me3 modification.More importantly,we observed the 3'-H3K4me3 enrichment among genes in human,fruitfly and Arabidopsis,and found that the sequences of 3'-H3K4me3-marked regions are highly conserved and essentially indistinguishable from known promoters in vertebrate.Therefore,we speculate that these 3'-H3K4me3-marked regions may serve as potential promoters for antisense transcription and 3′-H3K4me3 appear to be a universal epigenetic feature in eukaryotes.Our results provide a novel insight into the epigenetic roles of H3K4me3 and the regulatory mechanism of antisense transcription.