目的:检测lncRNA-DNAJC3-AS1在子宫颈鳞癌组织中的表达水平及其临床意义.方法:运用RNA原位杂交技术检测lncRNA-DNAJC3-AS1在73例宫颈鳞癌和癌旁正常组织中的表达情况,并分析lncRNA-DNAJC3-AS1与宫颈鳞癌患者病理资料和预后生存的相关性.结果:lncRNA-DNAJC3-AS1在宫颈鳞癌组织中呈阳性表达,约占83.56％(61/73);与癌旁正常组织阳性表达(5.48％,4/73)之间有统计学差异(χ2=90.096,P<0.05).高表达lncRNA-DNA-JC3-AS1的例数为57例,低表达lncRNA-DNAJC3-AS1的例数为16例.高表达的lncRNA-DNAJC3-AS1与宫颈鳞癌的分化程度和淋巴结转移密切相关(P<0.05),而与患者年龄、HPV感染、肿瘤大小和TNM分期无相关性(P>0.05).低表达组的生存时间明显长于高表达组(χ2=33.85,P<0.05).结论:lncRNA-DNAJC3-AS1可能是宫颈鳞癌的一种新的细胞癌基因,可以作为宫颈鳞癌发生和预后判断的新标志物;lncRNA-DNAJC3-AS1促进了宫颈鳞癌的发生发展过程.

Pigeon paramyxovirus type 1 (PPMV-1) is considered an antigenic and variant of avian paramyxovirus type 1 (APMV-1) that has adapted to pigeons as hosts. However, how this host-specific adaption of PPMV-1 is related to its biological characteristics is unknown. In this study, seven unique amino acids in PPMV-1 that are not present in other APMV-1 strains (n = 39 versus n = 106) were identified. R36 of the M protein was found to be not only a unique amino acid but also a positive-selection site. To investigate the role of R36 in host adaptation, a recombinant PPMV-1 with R36Q mutation was constructed. Our results indicated that the an R36Q mutation significantly attenuates pathogenicity in chickens, viral growth in both chicken embryo fibroblasts (CEFs) and pigeon embryo fibroblasts (PEFs), and virus replication and shedding in pigeons in comparison with the wild-type virus, suggesting that R36 is a key residue that evolved during the adaptation of PPMV-1 in pigeons.

In the process of modernization of the fast development,with some minority region way,way of working,the contact method of change,the native folk belief system lost past the ecological environment,caused many folk faith and form distortion and variation,cause for inheritance of a narrow and be forgotten.This paper takes Gui zhou province's qiandongnan Carey three trees town sword village folk beliefs season and the form of inheritance and protection of current situation and the problems of this issue as an example and puts forward related suggestions.

Spleen is the target organ of Newcastle disease virus(NDV).To elucidate the interaction between NDV and its hosts,and to further evaluate the pathological differences of spleen from the geese infected by the early-genotype NDV strain Herts/33(IV) and recent-genotype NDV strain JS5/05(VIId),the comparative proteomes of spleen from geese infected with these two NDVs were analyzed using two-dimensional gel electrophoresis(2-DE).Thirty-six 30-day-old geese were divided into three groups.The spleens from the geese in different groups were collected at 36 hours,72 hours and 108 hours post infection.The soluble proteins were extracted and then separated with 17 cm IPG strips(pH 5-8).Moreover,the 2-DE maps were comparatively analyzed by the software PDQuest 8.0.1.Compared with the control group,154 and 148 differentially expressed protein spots were found in Herts/33-and JS5/05-infected group,respectively.There were 86 spots shared by the two NDV-infected groups,with 52 proteins up-regulated and 34 proteins down-regulated.In addition,a total of 130 differentially expressed protein spots were identified between Herts/33-and JS5/05-infected groups,including 71 up-regulated proteins and 59 down-regulated proteins.The virulent genotype Ⅳ and genotype Ⅶd NDVs induced distinct splenic protein expression profiles after infection,which provided useful information to further elucidate the molecular mechanisms of increased pathogenicity of genotype Ⅶd NDV to waterfowl.

归渡,是中国山水画中的常见题材.“舟”作为古时人们重要的交通工具,她寄托的是归家人的一份情怀,其中掺杂着无奈、等待和焦急的复杂心情.纵观美术史,这一题材的出现有着深刻的文化根源和社会根源,它的出现和发展有着特殊的意义.自六朝至宋元,归渡题材的山水画从未间断,同时在笔墨技巧方面又有所创新,可以看出,归渡题材的山水画中饱含着文人骚客的无限情感.

Background Six nucleotide (nt) insertion in the 5'-noncoding region (NCR) of the nucleoprotein (NP) gene of Newcaslte disease virus (NDV) is considered to be a genetic marker for recent genotypes of NDV, which emerged after 1960. However, F48-like NDVs from China, identified a 6-nt insert in the NP gene, have been previously classified into genotype III or genotype IX. Results In order to clarify their phylogenetic position and explore the origin of NDVs with the 6-nt insert and its significance in NDV evolution, we determined the entire genome sequences of five F48-like viruses isolated in China between 1946 and 2002 by RT-PCR amplification of overlapping fragments of full-length genome and rapid amplification of cDNA ends. All the five NDV isolates shared the same genome size of 15,192-nt with the recent genotype V-VIII viruses whereas they had the highest homology with early genotype III and IV isolates. Conclusions The unique characteristic of the genome size and phylogenetic position of F48-like viruses warrants placing them in a separate geno-group, genotype IX. Results in this study also suggest that genotype IX viruses most likely originate from a genotype III virus by insertion of a 6-nt motif in the 5'-NCR of the NP gene which had occurred as early as in 1940 s, and might be the common origin of genotype V-VIII viruses.

Seventy-nine Newcastle disease viruses (NDV) isolated from clinical specimens of different poultry species including chickens, pigeons (Columba livia), geese and ostriches in Eastern China during 2005–2008 were characterized biologically and phylogenetically. The results showed genetic diversity of these viruses: three class I viruses and one genotype I and 12 genotype II viruses of class II circulating in chickens were avirulent; four genotype VIb viruses isolated from pigeons were moderately virulent; and two genotype III viruses and 57 genotype VIId viruses were highly virulent. The three class I viruses were further classified as genotypes 2 and 3. The very high F protein sequence identity of one genotype I virus with strain Queensland V4 and 12 genotype II viruses with strain La Sota indicated that these viruses originated from the two vaccine strains. Two genotype III viruses shared greater than 99% sequence identity with the moderately virulent vaccine strain Mukteswar but exhibited significantly higher virulence, suggesting that they evolved from the vaccine virus and that the Mukteswar vaccine should be banned in China. Fifty-seven of the 63 virulent NDVs in this study belonged to genotype VIId, indicating its predominance in Eastern China. Genotype VIId viruses could be further classified into two subgroups. Four of the five NDVs isolated from pigeons belonged to genotype VIb, indicating its host-specific preference. Both the genotype VIb and VIId NDVs showed low amino acid similarity to the vaccine strains currently used in China, implying the urgent need to develop better vaccines against the most prevalent NDVs in China.

With Wu Zhen's "Pine & Springs Painting" as the breakthrough points, this paper elaborates on the integration of poem and painting, introducing the close relationship between poem,painting and scene. Finally, it comes to the following conclusion:only the integration of subjective and objective as well as affection and scene and the succession of the predecessors' experience can help produce an art composition with both mood and affection.

To establish a stable technique of two-dimensional gel electrophoresis(2-DE) for chicken embryo fibroblasts (CEF) infected with Newcastle disease virus(NDV),cell culture,sample preparation,loading capacity,isoelectric focusing,staining and other steps were optimized using NDV-infected CEF as model system. About 800 protein spots on 17 cm IPG strip(pH 4 to 7) stained with blue sliver were analyzed by the software PDQuest8.0.1,and the match rate of protein spots was over 90%. The analysis of multiple 2-DE gels revealed that a total of 36 protein spots differentially expressed during NDV infection,including 16 up-regulated protein spots,15 down-regulated protein spots,3 new emerging protein spots and 2 disappear protein spots. Those differentially expressed protein spots were useful to further investigation of the interaction between NDV and its hosts.

In order to investigate the molecular epidemiology of Newcastle disease virus in China,forty-three Newcastle disease viruses(NDV) isolated from Eastern China during 2006-2008 were characterized phylogeniclly.Genetic analysis based on the HN gene sequences revealed that the isolates were classified into four genotypes: 2 isolates belonged to genotype I,3 to genotype Ⅱ,1 to genotype Ⅵ and 37 to genotype Ⅶd.The genotype Ⅶd NDVs could be further classified into two subgroup,Ⅶd 1and Ⅶd 2.Compared of Ⅶd 1 viruses,the viruses in Ⅶd 2 presented the three amino acid sequence mutations of T102I,A118E and T443M in HN protein.Besides,the results showed that the separation rate of the E347K mutation isolation in the HN protein linear epitope in China is increasing with time.

To evaluate the pathogenicity of Newcastle disease virus(NDV) strains of different genotypes in pigeons,JS-7-05-Ch,WX-10-07-Pi,JS-5-05-Go and F48E8 NDV strains that belonged to genotype Ⅲ,Ⅵ,Ⅶ and Ⅸ,respectively,were chosen to artificially infect two-month-old pigeons.Clinical symptoms and pathological changes of pigeons were observed,and humoral antibody response,virus shedding and virus distribution in tissues were examined.The results showed that NDVs could cause 100 % morbidity in pigeons and the mortality of pigeons infected with genotype Ⅲ-,Ⅵ-,Ⅶ-and Ⅸ-infected groups was 0,0,100 % and 50 %,respectively.Pigeons infected with WX-10-07-Pi experienced longer time in virus shedding and showed higher ratio of virus isolation when compared with other pigeons,which indicated that the Ⅵb NDVs might have genetic advantages over other genotypes in NDV outbreaks in pigeons.

Pair of primers and a TaqMan probe were synthesized according to M gene conservative sequence of Newcastle disease virus.The positive recombinant plasmid containing M gene of NDV ZJ1 strain isolated from goose was used as a positive quantitative template to establish a standard curve.And then a rea1-time fluorescent quantitative RT-PCR assay was established.The method has a good linear relationship within the 106 to 101 copies,with which 3 copies·μL-1 of the virus nucleic acid can be detected in the initial template,and has similar sensitivity with traditional virus isolation methods.The conforming rate of positive sum and negative sum with traditional virus isolation method was 90.0% and 99.8% respectively in detecting 500 clinic cloacal swab samples.The result showed that the constructed method paved the way for the early and rapid detection of NDV as well as quantitative analysis for the infection degree of NDV.

Eleven avirulent Newcastle disease viruses(NDVs) were isolated from ducks in China from 2002 to 2007.Phosphoprotein(P) genes of these NDVs were amplified by RT-PCR and sequenced.The P gene sequences were analyzed using bioinformatic software(SNAP and CODEML) to examine genetic variability and their effect on virus evolution.The P gene sequences of twenty eight NDV reference strains in GenBank were also included for comparison in the present study.Phylogenetic analysis of the P gene indicated that Class I genotype 2 Chinese viruses were closely related to some European and American viruses and Class I genotype 3b viruses formed a separate branch,which geographically distributed within China only.Class II genotype Ib viruses diverged from the representative strains of genotype I and the vaccine virus Queensland V4,the latter is extensively used in China.The bioinformatic analysis revealed graphic patterns of the dS and dN values and confirmed the prevalence of positive selection at terminal regions of P proteins(aa 62-113 and 137-198).Class I genotype 3b virus was unique to China.The P protein of different genotypes was most highly variable and the variability was presented in a few localized regions under negative and positive selections.

We isolated and identified 201 Newcastle disease viruses (NDVs) from domestic ducks in a 5-year surveillance study at live bird markets in Eastern China. Seventy-three of these isolates were characterized biologically and genetically. Fusion protein (F) genes of these isolates were amplified by reverse transcription-polymerase chain reaction and sequenced. Intracerebral pathogenicity index tests in 1-day-old specific-pathogen-free chickens and the mean death time of embryonated fowl eggs in addition to the cleavage site analysis of the F-protein precursor for these viruses showed that they were all avirulent NDVs. Phylogenetic analysis based on partial sequences of the F gene showed that 30 isolates clustered into the class I clade and the other 43 isolates clustered into genotype I of class II, but diverged from the vaccine virus Queensland V4, which is extensively used in China. Most class I viruses (18/30) formed a separate branch closest to the Hong Kong live bird market strains that have been recently designated as genotype 3, while the rest (12/30) were closely related to some European viruses within genotype 2. All of the 43 class II genotype I viruses diverged from viruses originally assigned to genotype Ia and formed a separate sublineage designated as Ib with water bird isolates from the Far East, suggesting the possible transmission between the wild and domestic waterfowl. The results in the present study clearly showed that the domestic duck population carries avirulent NDVs with genetic divergence regularly and may act as one of the important reservoirs.

To explore the molecular features of Class Ⅰ Newcastle disease viruses (NDV) and the role of NDV evolution during 2002 to 2007 in China,we andlyzed nucleoprotein (NP) gene sequences of eight ClassⅠ NDV. The NP gene sequences of twenty-six NDV reference strains in GenBank were also used in this study. The NP gene sequences were evaluated using bioinformatic software(DNAStar and MEGA4). Phylogenetic analysis of NP gene indicated that Class Ⅰ genotype 2 viruses were closely related to some European and American viruses within ClassⅠ;that ClassⅠ genotype 3 viruses diverged from other ClassⅠ viruses. It was found that NP protein was relatively conserved,especially in 1 to 400 amino acids,and amino acid substitutions were major in a region at P proteins (400 to 480). ClassⅠ genotype 2 and 3 viruses had different origins. The NP protein of different genotypes was relatively conserved,but in carboxylic- terminal (C-terminal) region of NP protein,amino acid sequence was highly variable,which may have the effect on its function.

OBJECTIVE:The purpose of this research is to establish a simple rapid amplification of cDNA ends (RACE) strategy for direct mapping of the 3' end and 5' end of the genomic RNA of Newcastle disease virus (NDV), and to analyze the leader and trailer sequence of NDV strains belonging to different genotypes.METHODS:Classic RNA Ligase Mediated Rapid Amplification of cDNA Ends (RLM-RACE) was specifically modified for mapping both ends of the NDV genome. 3'-RACE was carried out by genomic RNA ligation with 5' end phosphated adaptor CL+, and the 5' end was obtained by first strand cDNA with adaptor CL+.RESULTS:A modified RLM-RACE strategy was established in this paper, which proved simple, low-cost, repetitive and could be specifically used to map genome ends of NDV. By using this method, the leader and trailer sequence of 5 NDV strains, termed JS/5/05/Go, JS/07/04/Pi, JS/07/16/Pi, JS/7/05/Ch and JS/9/05/Go, belonging to genotype III, VI and VII was determined, respectively.CONCLUSION:The initial 8nt at the 3' and 5' ends of the genome of genotype I-VI NDV strains were complementary, whereas, the complementary sequences of strain JS/5/05/Go were up to 9 nt due to a mutation from T to C at the 9th nt in the 5' end. The 3' end of NDV genomic and anti-genomic RNA was predicted to form a potential hairpin structure. The U-->C(T-->C) mutation was located in the circle part of the hairpin in the 5' end of anti-genomic RNA, and had no visible influence on the formation of RNA secondary structure. However, the sequence of the circle part of the hairpin was changed from 3'-UUUC-5' to 3'-UCUC-5', more similar to the 3'-UCUUA-5' in the hairpin of genomic RNA.

The biological properties of six chicken-origin H9N2 subtype avian influenza viruses(AIVs) isolated from eastern China were characterized and the molecular phylogenetic analysis of HA and NA genes was made.All these viruses were non-pathogenic to SPF chicken with IVPI scores of 0,and grouped into two subgroups by monoclonal antibody of 4E7 with hemagglutination inhibition(HI) test.The sequence analysis of HA gene showed that the amino acid sequence in HA cleavage motif was RSSR↓GLF in all six strains,and the amino acid residues of potential glycosylation sites in isolates of Ck/HD/16/07 and Ck/HD/112/05 were different from those of other four isolates.The sequence analysis of NA genes indicated that NA gene was assigned to N2 subtype,and NA gene in four isolates deleted three amino acid residues in the stalk,whereas there was no deletion in other two isolates.

Twenty Newcastle disease virus(NDV)strains were isolated from diseased chicken and geese in field outbreaks during 2005 and 2006 in some regions of Jiangsu and Guangxi,and the antigenic analysis of the all NDV isolates had been done based on the reaction spectrum with a panel of monoclonal antibodies to the HN glycoprotein.The entire ORFs encoding HN protein of these NDV isolates were amplified by RT-PCR successfully,cloned and sequenced.The resultant sequences of HN genes of 13 isolates of chicken origin and 7 isolates of goose origin were gained and analyzed.The results of reaction spectrum showed that there were some distinct differences in the antigenic epitopes among the 20 NDV isolates.And the sequences revealed that the coding regions of the HN genes of these isolates all consisted of 1716 nt characteristic of virulent strains of NDV,coding for 571 amino acids.Neucleotides sequence homology were found to be from 94.8%to 100%among 18 NDV isolates of genotypeⅦ,and the neucleotides sequence homology between all the isolates and the other genotypeⅦstrains of recent years in China ranged from 92.1%to 99.6%.The deduced amino acid sequences and the receptor-binding regions of HN proteins between the NDV isolates of chicken origin and of goose origin were compared and analyzed.The results showed that some unique amino acid substitutions were found in the genome of the NDV isolates,and the close genetic similarity provided evidence for epidemiological linkage between the NDV isolates of chicken origin and of goose origin in the same period.

Eighty-four Salmonella enterica isolates were collected from chickens with clinical signs. 41 of the 84 isolates(49%) were found to be resistant to tetracycline. Of the 41 tetracycline-resistant isolates,the tetracycline-resistant genes were detected by PCR amplification as follows:tet(A)gene was the sole tetracycline-resistant gene found in all the isolates of S. Pullorum and S. Gallinarum;among the strains of S. Enteritidis and S. Derby,8 strains carried a tet(A)gene,17 carried a tet(B),and 10 carried a tet(G)gene. No tet(C) gene was detected in all 41 tetracycline-resistant isolates. The tetracycline-resistant genes located in the conjugation plasmids and were not related to gene cassettes.