Complete Nucleotide Sequence of a Mumps Virus SP Strain Isolated in China

The complete nucleotide sequence of the mumps virus SP, which was isolated in China, was determined. As with other mumps viruses, its genome was 15 384 nucleotides (nts) in length and encoded seven proteins. The full-length nucleotide sequence of the SP isolate differed from other strains by 4% –6.8% at the nucleotide sequence level. Due to variations of amino acids over the full genome (including the HN and N genes), this isolate exhibited significant variations in the antigenic sites. This report is the first to describe the full-length genome of a genotype F strain and provide an overview of the diversity of genetic characteristics of a circulating mumps virus.

Isolation and Complete Nucleotide Sequence of the Measles Virus IMB-1 Strain in China

The complete nucleotide sequence of the measles virus strain IMB-1,which was isolated in China,was determined.As in other measles viruses,its genome is 15,894 nucleotides in length and encodes six proteins.The full-length nucleotide sequence of the IMB-1 isolate differed from vaccine strains (including wild-type Edmonston strain) by 4%-5% at the nucleotide sequence level.This isolate has amino acid variations over the full genome,including in the hemagglutinin and fusion genes.This report is the first to describe the full-length genome of a genotype H1 strain and provide an overview of the diversity of genetic characteristics of a circulating measles virus.

Characterizing Long-range Correlation Properties in Nucleotide Sequences

Using continuous wavelet transform as the analytical tool, the fractal characteristic of nucleotide sequences was studied. The fractal dimension of the exon and intron sequences for different species was calculated. We use the Mexican hat wavelet function as the mother wavelet and Hurst exponent to describe the long-range correlation. It is found that the Hurst exponent of intron sequence is larger than that of exon sequence for the same gene.

KoNA:Korean Nucleotide Archive as A New Data Repository for Nucleotide Sequence Data

During the last decade,the generation and accumulation of petabase-scale high-throughput sequencing data have resulted in great challenges,including access to human data,as well as transfer,storage,and sharing of enormous amounts of data.To promote data-driven biological research,the Korean government announced that all biological data generated from government-funded research projects should be deposited at the Korea BioData Station(K-BDS),which consists of multiple databases for individual data types.Here,we introduce the Korean Nucleotide Archive(KoNA),a repository of nucleotide sequence data.As of July 2022,the Korean Read Archive in KoNA has collected over 477 TB of raw next-generation sequencing data from national genome projects.To ensure data quality and prepare for international alignment,a standard operating procedure was adopted,which is similar to that of the International Nucleotide Sequence Database Collaboration.The standard operating procedure includes quality control processes for submitted data and metadata using an automated pipeline,followed by manual examination.To ensure fast and stable data transfer,a high-speed transmission system called GBox is used in KoNA.Furthermore,the data uploaded to or downloaded from KoNA through GBox can be readily processed using a cloud computing service called Bio-Express.This seamless coupling of KoNA,GBox,and Bio-Express enhances the data experience,including submission,access,and analysis of raw nucleotide sequences.KoNA not only satisfies the unmet needs for a national sequence repository in Korea but also provides datasets to researchers globally and contributes to advances in genomics.The KoNA is available at https://www.kobic.re.kr/kona/.

Isolation and Complete Genomic Sequence Analysis of a New Sindbis-like Virus

The Sindbis-like virus was first discovered in China in 1986. Its complete genomic sequence consists of more than 11 000 bp encoding more than 3 700 amino acids. It contains a 5＇ non-transcriptional region （5＇-NTR） in a non-structural region, four non-structural proteins （nsP1, nsP2, nsP3, nsP4） regions, capsids in conserved and non-conserved regions and structural El, E2, E3, 6K regions and a 3＇ non-transcriptional region （3＇-NTR）. The Sindbis-IMB was isolated from the blood of a patient suspected to have encephalitis, and was followed by identification and passage. The virus RNA was extracted from virus supernatant in infected cells and the whole genome was divided into 12 fragments; RT-PCR was then performed to amplify the 12 fragments for complete sequencing. The results showed that the whole genomic sequence of Sindbis-IMB consists of 11 717 bp encoding 3 773 amino acids. Homology comparison with other Sindbis-like isolates demonstrated the highest similarity was the YN87448 with a variation of 1% strain isolated in Yunnan Province and the second highest to the SAAR86 strain with a variation of -1.2%. The nucleotide sequence variations were present in non-structural regions, resulting in amino acids K, E, N, R, H, and L in protein sequences in positions 230, 231,443,781, 1 582, and 1746 in the new isolation respectively. Furthermore, three additional amino acids-glutamic acid, serine and alanine-were noted in nsp4 terminus as compared to the YN87448 isolate

Cloning and Sequence Analysis of Prophenoloxidase from Haemocytes of the Red Swamp Crayfish,Procambarus clarkii

The full-length cDNA sequence of prophenoloxidase was obtained through RACE technology. The complete cDNA sequence is 3 721-bp long, containing an open reading frame （ORF） of 1 881 bp, a 154-bp 5′-untranslated region, and a 1 686- bp 3′-untranslated region with three potential functional poly（A） signals （AATAAA）. The molecular mass of the deduced amino acid sequence （627 aa） was 72.3 kDa with an estimatedpI of 5.88. It contained putative copper-binding sites （copper A： 131, 135, 167 and copper B： 301,305, 341）, and a tentative complement-like motif （GCGWPDHL）. Eight potential N-linked glycosylation sites were predicted to be present in P. clarkii prophenoloxidase. Similar to those in other arthropod prophenoloxidases reported so far, no signal peptide was detected in the crayfish prophenoloxidase. The phylogenetic trees confirmed that P. clarkii prophenoloxidase was most closely related to that of freshwater crayfish P. leniusculus and more closely related to other crustacean prophenoloxidases from shrimp, prawn, and lobster than to the insect prophenoloxidases. Besides, two putative introns were found in this sequence of genomic DNA.

Cloning and sequence analysis of β-actin gene from Aedes albopictus (Diptera: Culicidae)

Objective： To obtain the complete β-actin gene from Aedes albopictus. Methods： Total RNA was extracted from C6/36 cells. Degenerate primers were designed based on the β-actin sequences of An. gambiae, Ae. aegypti, Cx. pipiens pallens and D. melanogaster. By RT-PCR, the product was amplified, purified, cloned into the pGT vector and sequenced. The β-actin sequence was aligned and phylogenetically analyzed by the BLAST program and the CLUSTAL W program. Results： A sequence of 1132 bp including an open reading frame of 1131 bp was obtained （GenBank DQ657949）. The deduced protein had 376 amino acids. Aligned to SWISS-PROT, it exhibited a high level of identity with β-actins from Anopheles, Drosophila and Culex at the amino acid sequence level. Phylogenetic analysis indicated that Ae. albopictus β-acfin was much more homologous with invertebrate β- actin than with vertebrate β-actin. Conclusion： The gene may be used as the internal control in the experiments of Ae. albopictus.

GenBase: A Nucleotide Sequence Database

The rapid advancement of sequencing technologies poses challenges in managing the large volume and exponential growth of sequence data efficiently and on time.To address this issue,we present GenBase(https://ngdc.cncb.ac.cn/genbase),an open-access data repository that follows the International Nucleotide Sequence Database Collaboration(INSDC)data standards and structures,for efficient nucleotide sequence archiving,searching,and sharing.As a core resource within the National Genomics Data Center(NGDC)of the China National Center for Bioinformation(CNCB;https://ngdc.cncb.ac.cn),GenBase offers bilingual submission pipeline and services,as well as local submission assistance in China.GenBase also provides a unique Excel format for metadata description and feature annotation of nucleotide sequences,along with a real-time data validation system to streamline sequence submissions.As of April 23,2024,GenBase received 68,251 nucleotide sequences and 689,574 annotated protein sequences across 414 species from 2319 submissions.Out of these,63,614(93%)nucleotide sequences and 620,640(90%)annotated protein sequences have been released and are publicly accessible through GenBase’s web search system,File Transfer Protocol(FTP),and Application Programming Interface(API).Additionally,in collaboration with INSDC,GenBase has constructed an effective data exchange mechanism with GenBank and started sharing released nucleotide sequences.Furthermore,GenBase integrates all sequences from GenBank with daily updates,demonstrating its commitment to actively contributing to global sequence data management and sharing.

Association of Single Nucleotide Polymorphisms of ABCB1,OPRM1 and COMT with Pain Perception in Cancer Patients

Pain perception is influenced by multiple factors. The single nucleotide polymorphisms（SNPs） of some genes were found associated with pain perception. This study aimed to examine the association of the genotypes of ABCB1 C3435 T,OPRM1 A118 G and COMT V108/158M（valine 108/158 methionine） with pain perception in cancer patients. We genotyped 146 cancer pain patients and 139 cancer patients without pain for ABCB1 C3435T（rs1045642）,OPRM1 A118G（rs1799971） and COMT V108/158M（rs4680） by the fluorescent dye-terminator cycle sequencing method,and compared the genotype distribution between groups with different pain intensities by chi-square test and pain scores between groups with different genotypes by non-parametric test. The results showed that in these cancer patients,the frequency of variant T allele of ABCB1 C3435 T was 40.5%; that of G allele of OPRM1 A118 G was 38.5% and that of A allele of COMT V108/158 M was 23.3%. No significant difference in the genotype distribution of ABCB1 C3435T（rs1045642） and OPRM1 A118G（rs1799971） was observed between cancer pain group and control group（P=0.364 and 0.578）; however,significant difference occurred in the genotype distribution of COMT V108/158M（rs4680） between the two groups（P=0.001）. And the difference could not be explained by any other confounding factors. Moreover,we found that the genotypes of COMT V108/158 M and ABCB1 C3435 T were associated with the intensities of pain in cancer patients. In conclusion,our results indicate that the SNPs of COMT V108/158 M and ABCB1 C3435 T significantly influence the pain perception in Chinese cancer patients.

Cloning and Sequence Analysis of Envelope Glycoprotein E1 Gene of Rubella Virus, JR23 Strain

To construct an expression vector containing the E1 glycoprotein gene of rubella virus for the study on the effect of mutation of the E1 gene glycoprotein and the analysis of phylogenetic differences of sequences, the gene encoding the E1 envelope glycoprotein was amplified from rubella virus, Jinan strain JR23, by RT-PCR and ligated into PMD-18T vector. The clones that carried the E1 gene were identified after amp r selection and analysis of restriction enzyme digestion. After sequencing this gene was analyzed by Danstar and Winstar programs, and the map of phylogenetic tree was drawn. The clone of E1 glycoprotein was thus constructed. It was found that the sequence differences between JR23 strain and the TCRB strain from Japan and those between JR23 strain and Thomas strain of England were rather small with difference values of 0.9% and 1.2% respectively. Yet those between JR23 strain and BRD2 strain from Beijing and those between JR23 strain and XG379 strain from Hong Kong were comparatively larger with difference values of 7.6% and 7.3% respectively. The sequence of JR23 strain with other strains was less than 3% except the NC strain (3.7%). It concludes that the construction of E1 glycoprotein gene offers an approach to study the relationship between structures and functions of E1 gene and its gene products. In the phylogenetic tree, it shows that there are significant differences in the sequences of rubella virus isolated in China, and this might be helpful to develop an effective subunit vaccine.

Video Transmission Secrecy Improvement Based on Fractional Order Hyper Chaotic System

In the Digital World scenario,the confidentiality of information in video transmission plays an important role.Chaotic systems have been shown to be effective for video signal encryption.To improve video transmission secrecy,compressive encryption method is proposed to accomplish compression and encryption based on fractional order hyper chaotic system that incorporates Compressive Sensing(CS),pixel level,bit level scrambling and nucleotide Sequences operations.The measurement matrix generates by the fractional order hyper chaotic system strengthens the efficiency of the encryption process.To avoid plain text attack,the CS measurement is scrambled to its pixel level,bit level scrambling decreases the similarity between the adjacent measurements and the nucleotide sequence operations are done on the scrambled bits,increasing the encryption.Two stages are comprised in the reconstruction technique,the first stage uses the intra-frame similarity and offers robust preliminary retrieval for each frame,and the second stage iteratively improves the efficiency of reconstruction by integrating inter frame Multi Hypothesis(MH)estimation and weighted residual sparsity modeling.In each iteration,the residual coefficient weights are modified using a mathematical approach based on the MH predictions,and the Split Bregman iteration algorithm is defined to resolve weighted l1 regularization.Experimental findings show that the proposed algorithm provides good compression of video coupled with an efficient encryption method that is resistant to multiple attacks.

Molecular Phylogeny Supports the Validity of Polypedates impresus Yang 2008

The taxonomy of the Asian tree frogs of the Polypedates leucomystax complex, a group of widespread and morphologically similar species, is very controversial. To ascertain the taxonomic status of these species, we investi- gated the historical relationships among representative samples based on - 2 kb of nucleotide sequences from the mito- chondrial 12S rRNA, tRNAvalinc, and 16S rRNA genes. Our phylogeny resolved five well supported lineages （A, B, C, D and E） in the P leucomystax complex. Polypedates impresus from Yunnan, China, Polypedates cf. mutus and Polypedates cf. megacephalus from Guangxi and Yunnan, China, and Laos constructed Clade C, which is monophyletic. In order to recognize the unique position of this clade, we considered P impresus in Clade C as a valid species. Following our phy- logeny, Chinese Polypedates, corresponding to the other four clades, should include four species： P. mutus （Clade A）; P. braueri （Clade B）; P impresus （Clade C） and P. megacephalus （Clade D）. P Ieucomystax （Clade E） is mainly distributed in the Malaysia, Indonesia and Philippines.

Genetic Characterization of Fusion Protein of Human Respiratory Syncytial Virus: Beijing

Objective Fusion protein is a subunit of the human respiratory syncytial virus(HRSV)and a potential vaccine candidate.Thus,a study on the genetic characteristics of F protein was considered important for further investigations in this field.The aim of this study was to determine the prevalence and genetic diversity of the F gene of HRSV infections in hospitalized pediatric patients in Beijing with acute lower respiratory tract infections and to compare the circulating genotypes that are currently found worldwide.Methods HRSV particles were amplified by RT-PCR and the PCR products were purified for sequencing.Further analysis was carried out by Bioedit and MEGA 3.0 biological software programs.Results Seventy-six samples(23.1%)were positive for HRSV.The percentage of cases in patients younger than 1year was 84.21%.Among the six Beijing isolates,four belonged to subgroup A,whose respective F genes shared97.0%-97.4%nucleotide sequence identity and 92.1%-93.0%amino acid sequence identity.The other two isolates belonged to subgroup B.Here,97.3%and 98.2%sequence identity were found at nucleotide and amino acid levels,respectively.Conclusions Phylogenetic analysis of nucleotide sequences revealed that those four isolates within subgroup A were monophyletic and closely related to each other,but those two within subgroup B distributed in two distinct clusters.Subgroup A and B strains co-circulated,indicating that two different transmission chains occurred in Beijing from 2003-2004.

Sequence Information on Simple Sequence Repeats and Single Nucleotide Polymorphisms through Transcriptome Analysis of Mungbean

Mungbean （Vigna radiata （L.） Wilczek） is a unique species in its ability to fix atmospheric nitrogen, with early maturity, and relatively good drought resistance. We used 454 sequencing technology for transcriptome sequencing. A total of 150 159 and 142 993 reads produced 5 254 and 6 374 large contigs （〉_500 bp） with an average length of 833 and 853 for Sunhwa and Jangan, respectively. Functional annotation to known sequences yielded 41.34% and 41.74% unigenes for Jangan and Sunhwa. A higher number of simple sequence repeat （SSR） motifs was identified in Jangan （1 630） compared with that of Sunhwa （1 334）. A similar SSR distribution pattern was observed in both varieties. A total of 8 249 single nucleotide polymorphisms （SNPs） and indels with 2 098 high-confidence candidates were identified in the two mungbean varieties. The average distance between individual SNPs was -860 bp. Our report demonstrates the utility of transcriptomic data for implementing a functional annotation and development of genetic markers. We also provide large resource sequence data for mungbean improvement programs.

Genetic evaluation and testing for hereditary forms of cancer in the era of next-generation sequencing

The introduction of next-generation sequencing(NGS) technology in testing for hereditary cancer susceptibility allows testing of multiple cancer susceptibility genes simultaneously. While there are many potential benefits to utilizing this technology in the hereditary cancer clinic, including efficiency of time and cost, there are also important limitations that must be considered. The best panel for the given clinical situation should be selected to minimize the number of variants of unknown significance. The inclusion in panels of low penetrance or newly identified genes without specific actionability can be problematic for interpretation.Genetic counselors are an essential part of the hereditary cancer risk assessment team, helping the medical team select the most appropriate test and interpret the often complex results. Genetic counselors obtain an extended family history, counsel patients on the available tests and the potential implications of results for themselves and their family members(pre-test counseling), explain to patients the implications of the test results(post-test counseling), and assist in testing family members at risk.

Mycobacterium tuberculosis bacteremia in a human immunodeficiency virus-negative patient with liver cirrhosis:A case report

BACKGROUND With the increasing prevalence of human immunodeficiency virus(HIV),the incidence of Mycobacterium tuberculosis(M.tuberculosis)bacteremia has also increased.As a common affliction of acquired immunodeficiency syndrome patients,M.tuberculosis infection is associated in these patients with severe sepsis and high mortality.In contrast,M.tuberculosis bacteremia is rarely seen in HIVnegative patients,and M.tuberculosis has never been reported from the blood of patients with liver cirrhosis.CASE SUMMARY We evaluated a 55-year-old Chinese male patient who had been admitted to the hospital with abdominal distension of unknown cause of one-week duration,accompanied by diarrhea,shortness of breath,and occasional fever.Based on these indicators of abnormal inflammation and fever,we suspected the presence of an infection.Although evidence of microbial infection was not found in routine clinical tests and the patient did not show typical clinical symptoms of infection with M.tuberculosis,next-generation sequencing of blood samples nevertheless demonstrated the presence of M.tuberculosis,which was subsequently isolated from blood samples grown in conventional Bac T/ALERT FA blood culture bottles.CONCLUSION Our findings demonstrate that HIV-negative liver cirrhosis patients can also be infected with M.tuberculosis.

Sequence analysis of a soil-borne wheat mosaic virus isolate from Italy shows that it is the same virus as European wheat mosaic virus and Soil-borne rye mosaic virus

The complete sequence of the two RNAs of a furovirus isolate fromdurum wheat in Italy was determined. Sequence comparisons and phylogenetic analysis were done to compare the Italian virus with Soil-borne wheat mosaic virus (SBWMV) from the USA and with furovirus sequences recently published as European wheat mosaic virus (EWMV), from wheat in France, and Soil-borne rye mosaic virus (SBRMV), from rye and wheat in Germany. Over the entire genome, the Italian isolate RNA1 and RNA2 had respectively 97.5% and 98.6% nucleotide identity with EWMV, 95.5% and 85.8% with SBRMV-G and 70.6% and 64.5% with SBWMV. The Italian isolate was therefore clearly distinct from SBWMV. The European isolates all appear to belong to the same virus and the name Soil-borne cereal mosaic virus may resolve earlier ambiguities.

High-throughput sequencing analysis of differential microRNA expression in the process of blocking the progression of chronic atrophic gastritis to gastric cancer by Xianglian Huazhuo formula(香连化浊方)

OBJECTIVE:To explore the mechanism of Xianglian Huazhuo formula(香连化浊方,XLHZ)blocking the development of chronic atrophic gastritis(CAG)to gastric cancer(GC)through bioinformatics analysis and in vitro.METHODS:Pathological morphology of gastric mucosa of rats were observed.High-throughput sequencing was used to analyze the miRNA expression profile of gastric mucosa.The miRanda,miRDB and miRWalk databases were used to predict the differential target genes.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were performed for differential target genes.Real-time quantitative reverse transcription polymerase chain reaction(qRTPCR)was used to verify the differentially expressed miRNAs and target genes.Western blot,EdU,wound healing and flow cytometry were used to observe the effect of XLHZ on epithelial-mesenchymal transition(EMT)markers,proliferation,migration,apoptosis and cell cycle of CAG cells in vitro.RESULTS:A total of five differentially expressed miRNAs and four differential target genes were screened in this study.GO analysis showed that the target genes were enriched in regulation of neuron development,regulation of transcription factor activity and regulation of RNA polymerase.KEGG pathways database differences in gene enrichment of target genes in the Wnt signaling pathway,Phospholipase D signaling pathway and mitogen-activated protein kinase signaling pathway.qRTPCR confirmed that miRNAs and its target genes were consistent with the screening results.In vitro,our study revealed that XLHZ could increase the expression of Ecadherin,decrease the expression of transforming growth factorβ1,vimentin andβ-catenin,inhibite the proliferation and migration of CAG cells,cause cell cycle arrest at G0/G1 and G2/M phase,induce the apoptosis of CAG cells,and prevent the progression of CAG to GC.CONCLUSION:This study provided a new idea for the mechanism of blocking the progression of CAG to GC by XLHZ,which may be related to the expression of miR-20a-3p,miR-320-3p,miR-34b-5p,miR-483-3p and miR-883-3p and their target genes transferrin receptor,nuclear receptor subfamily 4 member 2,delta like canonical Notch ligand 1 and a kinase anchor protein 12 in CAG.In the future,we will continue to investigate the linkage between the active ingredients of XLHZ and the relevant miRNAs and their target genes,so as to provide more sufficient experimental basis for clinically effective prevention of CAG to GC.

The complete genome of Enterovirus 71 China strain

Five overlapping clones covering the full genome of Enterovirus71 China strain SHZH98 were obtained and then the sequences were determined by the chain termination method. It showed that the full length of EV71 SHZH98 genome (not including Poly A tail) is 7408 bp. There are some diversities on the lengths and sequences of 5′ UTR and 3′ UTR between SHZH98 and the other EV71 strains. In P1 capsid region, which is closely associated with viral immunogenicity, EV71 strain SHZH98 shares the highest homology with Taiwan strains; but in P2 and P3 non-structural gene regions there are higher identities with Coxsakievirus A16 and EV71 strains MS, BrCr than with Taiwan strains. Phylogenetic tree constructed by structural gene region indicates that China strain SHZH98 has a closer relationship with Taiwan strains, however, in the non-coding region it has a closer relationship with Coxsakievirus A16, EV71 strains MS and BrCr. EV71 China strain was analyzed at the molecular level. The results will contribute to the basic study on enteroviruses and the EV71 prevention in China.

Clinical efficacy of a combination of Percoll continuous density gradient and swim-up techniques for semen processing in HIV-1 serodiscordant couples

To evaluate the clinical efficacy of a procedure comprising a combination of Percoll continuous density gradient and modified swim-up techniques for the removal of human immunodeficiency virus type 1 （HIV-1） from the semen of HIV-1 infected males, a total of 129 couples with an HIV-1 positive male partner and an HIV-1 negative female partner （serodiscordant couples） who were treated at Keio University Hospital between January 2002 and April 2012 were examined. A total of 183 ejaculates from 129 HIV-1 infected males were processed. After swim-up, we successfully collected motile sperms at a recovery rate as high as 100.0% in cases of normozoospermia （126/126 ejaculates）, oligozoospermia （6/6）, and asthenozoospermia （36/36）. The recovery rate of oligoasthenozoospermia was 86.7% （13/15）. In processed semen only four ejaculates （4/181：2.2%） showed viral nucleotide sequences consistent with those in the blood of the infected males. After using these sperms, no horizontal infections of the female patients and no vertical infections of the newborns were observed. Furthermore, no obvious adverse effects were observed in the offspring. This protocol allowed us to collect HIV-1 negative motile sperms at a high rate, even in male factor cases. We concluded that our protocol is clinically effective both for decreasing HIV-1 infections and for yielding a healthy child.