China ; Genotype ; Measles virus ; genetics ; isolation & purification ; Molecular Epidemiology ; methods ; Nucleoproteins ; genetics ; Polymerase Chain Reaction

OBJECTIVETo investigate the impact of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) on sperm DNA fragmentation and nucleoprotein transition.

METHODSBased on the recommended methods in the WHO Laboratory Manual for the Examination and Processing of Human Semen (5th ed), we conducted routine semen analysis for 65 CP/CPPS patients and 30 healthy men. We also analyzed the results of papanicolaou staining, sperm DNA fragmentation and sperm nucleoprotein transition.

RESULTSCompared with the healthy control males, the CP/CPPS patients showed significant decreases in sperm concentration ([134.05 +/- 99.80] vs [94.75 +/- 92.07]) x 10(6)/ml, P <0.05), the percentage of morphologically normal sperm ([7.26 +/- 2.28] vs [5.61 +/- 3.40]%, P <0.05) and sperm progressive motility ([59.18 +/- 16.06] vs [47.68 +/- 17.62]%, P<0.05), but dramatic increases in sperm DNA fragmentation ([22.92 +/- 11.51] vs [43.58 +/- 17.07%, P<0.01) and sperm nucleoprotein transition ([23.26 +/- 5.97] vs [32.14 +/- 8.79]%, P<0.01).

CONCLUSIONCP/CPPS significantly reduces sperm quality and male fertility.

Adult ; Case-Control Studies ; DNA Fragmentation ; Humans ; Male ; Nucleoproteins ; genetics ; Prostatitis ; genetics ; Semen Analysis ; Sperm Count ; Sperm Motility ; Young Adult

This study aims to investigate whether the nucleoprotein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV) can impact the cellular immunity of host cells. Gene segments that encode the NP and non-structural protein (NSs) of SFTSV were inserted into eukaryotic expression vector VR1012. Host proteins that interact with NP and affect immunity were identified with co-immunoprecipitation (IP), SDS-PAGE, mass spectrometry (MS), and Western blot. Co-localization of NP and the identified host proteins was confirmed by confocal microscopy. A 60kD SSA/Ro, a protein related to immunity, interacted with NP, as found by IP and MS. Confocal microscopy showed that NP and SSA/Ro were co-localized in cytoplasm. These results indicated that SFTSV NP may specifically bind to 60kD SSA/Ro and cause a series of immune responses and clinical symptoms.
Bunyaviridae Infections ; genetics ; metabolism ; virology ; HEK293 Cells ; Humans ; Nucleoproteins ; genetics ; metabolism ; Phlebovirus ; genetics ; metabolism ; Protein Binding ; Ribonucleoproteins ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism

Ebola virus (EBOV) is a key member of Filoviridae family and causes severe human infectious diseases with high morbidity and mortality. As a typical negative-sense single-stranded RNA (-ssRNA) viruses, EBOV possess a nucleocapsid protein (NP) to facilitate genomic RNA encapsidation to form viral ribonucleoprotein complex (RNP) together with genome RNA and polymerase, which plays the most essential role in virus proliferation cycle. However, the mechanism of EBOV RNP formation remains unclear. In this work, we solved the high resolution structure of core domain of EBOV NP. The polypeptide of EBOV NP core domain (NP(core)) possesses an N-lobe and C-lobe to clamp a RNA binding groove, presenting similarities with the structures of the other reported viral NPs encoded by the members from Mononegavirales order. Most strikingly, a hydrophobic pocket at the surface of the C-lobe is occupied by an α-helix of EBOV NP(core) itself, which is highly conserved among filoviridae family. Combined with other biochemical and biophysical evidences, our results provides great potential for understanding the mechanism underlying EBOV RNP formation via the mobility of EBOV NP element and enables the development of antiviral therapies targeting EBOV RNP formation.
Crystallography, X-Ray ; Ebolavirus ; physiology ; Humans ; Nucleoproteins ; chemistry ; genetics ; metabolism ; Protein Structure, Tertiary ; Structure-Activity Relationship ; Virus Assembly ; physiology

OBJECTIVEThis study was to explore the differences in the nucleoprotein gene between rabies virus (RABV) and its vaccine strains in Guizhou province from year 2005 to 2010.

METHODSSamples from 4 rabies patients and cerebral tissue samples of 28 rabies infected dogs were collected from different districts in Guizhou province between year 2005 and 2010. Direct Immunofluorescence Assay (DFA) and RT-nested PCR assay were applied to detect the overall length of N gene sequence. Meanwhile, based on the comparison between the homology and phylogenetic tree, the differences in N gene sequence between the prevalent RABV and the RABV vaccine strains collected from NCBI database in these years.

RESULTSAccording to DFA and RT-nested PCR assay, the antigen and nucleic acid of the 21 dogs and 4 human samples were both confirmed positive; whose full length of N gene sequences were both 1353 bp. The homological analysis showed that the 25 strains of RABV virus and the RABV type I virus stored by GenBank database shared a high homology in N gene nucleotide and amino acid sequences, which were 89%-100% and 98%-100%, respectively. Besides, the homology between the 25 strains of RABV virus and its vaccines in nucleotide and amino acid sequences were separately 86%-95% and 96%-100%. The N gene of vaccines for livestock shared the highest homology with HEP-Flury strain in the nucleotide and amino acid, which were 88%-89% and 98%-99%, respectively. The vaccines for human use showed its greatest homology with the CTN strain in nucleotide (86%-100%) and amino acid (96%-100%). The phylogenetic tree analysis indicated that the 25 strains of RABV virus, RABV type I virus and the CTN vaccine strains constituted one individual cluster, which was least different from the CTN vaccine for human use.

CONCLUSIONThe prevalent RABV virus, the vaccine HEP-Flury for livestock and the vaccine CTN for human use were found to be highly similar in N gene expression in Guizhou province from 2005 to 2010.

Amino Acid Sequence ; Animals ; Dogs ; Genotype ; Humans ; Molecular Sequence Data ; Nucleoproteins ; genetics ; RNA, Viral ; genetics ; Rabies ; veterinary ; virology ; Rabies Vaccines ; genetics ; Rabies virus ; classification ; genetics ; isolation & purification

OBJECTIVETo study the genetic characterization of wild-type measles viruses isolated in Xinjiang in 2003 and 2004.

METHODSPeripheral blood mononuclear cells (PBMC) from 19 suspected measles cases collected between June 2003 and April 2004 were used to isolate measles virus by cocultivation with phytohemagglutinin (PHA)-stimulated cord blood mononuclear cells (CBMC). For positive samples, 676 nucleotides of the C-terminus of the nucleoprotein (N) gene of the measles virus genome were amplified by reverse transcription-polymerase chain reaction and then sequenced. These sequences were compared with those of other measles reference strains available in GenBank or measles isolates elsewhere in China using BLAST searches and phylogenetic analyses.

RESULTS6 measles virus strains were isolated with 3 strains (XJ03-26, XJ03-27, XJ03-74) from 2003 and 3 (XJ04-146, XJ04-150, XJ04-152) from 2004. The strain XJ03-26, differed from the Chinese measles vaccine strain S-191 (genotype A) by less than 1% at nucleotide level, and therefore appeared a vaccine-associated strain. The other 5 strains as XJ03-27, XJ03-74, XJ04-146, XJ04-150 and XJ04-152 were proved to be genotype H1 strains,among which XJ03-27, XJ03-74, XJ04-150 and XJ04-152, showing their nucleotide diversity were varied from 0.5% to 1.6%, when compared to the H1a reference strain China9322, and identified as H1a strains. XJ04-146 showed a nucleotide similarity of 98.7% when compared to H1b reference strain China9475, and was identified as H1b strain. Additionally, we found that there were two sets of strain (XJ03-27 and XJ04-150; XJ03-74 and XJ04-152), with almost identical nucleotide sequences, circulating in 2003 and 2004 and both having more nucleotide variability (up to 6.1%, 27 nucleotides).

CONCLUSIONGenotype H1 measles virus had been proven to have been circulated in Xinjiang, China during 2003 and 2004. H1a was the predominant epidemic strain while H1b strain stood the next.

China ; epidemiology ; Disease Outbreaks ; Genotype ; Humans ; Measles ; epidemiology ; Measles virus ; classification ; genetics ; isolation & purification ; Nucleoproteins ; genetics ; Phylogeny ; RNA, Viral ; genetics ; Sequence Analysis, RNA ; Viral Proteins ; genetics

Paramyxovirus Tianjin strain is a novel strain of virus causing common cotton-eared marmoset fatal infection. To investigate the relationship between the gene structure and function of nucleoprotein (NP) of Tianjin strain, NP gene of paramyxovirus Tianjin strain was cloned and three domains of NP were expressed. The homologous and phylogenetic analysis of NP sequences among the paramyxovirus Tianjin strain and eight strains of Sendai viruses from GenBank were performed. The results indicated the recombinant proteins NP1, NP2 and NP3 showed the native antigenicity to the polyclonal antiserum of paramyxovirus Tianjin strain, ranking as NP3>NP1>NP2 (precedence order). The homology of NP nucleotide and the deduced amino acid sequences between paramyxovirus Tianjin strain and Sendai virus BB1 strain were 94.5%, 96.2%, respectively, whereas the identity were 85.1% - 88.7% and 92.4% - 94.7% among Tianjin strain and the 7 strains of Sendai viruses from GenBank respectively. There were 15 unique amino acid substitutions in Tianjin strain NP protein and 11 common amino acid substitutions same with BB1 strain. This research confirmed that paramyxovirus Tianjin strain might be a new genotype of Sendai virus and can be helpful in the establishment of detection assay applying recombinant NP as antigen instead of the whole virions.
Amino Acid Sequence ; Blotting, Western ; Molecular Sequence Data ; Nucleoproteins ; classification ; genetics ; metabolism ; Paramyxovirinae ; genetics ; metabolism ; Phylogeny ; Recombinant Proteins ; metabolism ; Sequence Homology, Amino Acid ; Viral Proteins ; genetics ; metabolism

OBJECTIVETo investigate the correlation of sperm DNA damage and sperm-nucleoprotein transition with acrosin activity and seminal parameters.

METHODSWe collected 535 semen samples, assessed sperm DNA damage by sperm chromatin dispersion test, and analyzed the correlation of sperm DNA damage and sperm-nucleoprotein transition with acrosin activity and seminal parameters according to the WHO criteria.

RESULTSStatistically significant differences were observed in sperm DNA damage among sperm-nucleoprotein transition, acrosin activity, sperm concentration and the percentage of grade a + b sperm (P < 0.01). Sperm DNA damage was positively correlated with age, sperm-nucleoprotein transition, sperm concentration and the percentage of grade d sperm (P < 0.01 or P < 0.05), but negatively correlated with acrosin activity (P < 0.001). Stepwise linear regression analysis demonstrated that age, sperm concentration, the percentage of grade d sperm, sperm-nucleoprotein transition and acrosin activity were independent variables related to the DNA fragmentation index (DFI). The abnormality rates of sperm-nucleoprotein transition, acrosin activity, sperm concentration and graded a + b sperm were significantly higher in the sperm DNA damage group (DFI > or = 30%) than in the normal control (DFI < 30%) (P < 0.01).

CONCLUSIONSperm DNA damage is closely related with sperm-nucleoprotein transition, acrosin activity and seminal parameters, which may become another important independent parameter for the evaluation of sperm quality.

Acrosin ; genetics ; Adult ; Chromatin ; DNA Damage ; DNA Fragmentation ; Humans ; Infertility, Male ; genetics ; Male ; Nucleoproteins ; genetics ; metabolism ; Sperm Count ; Sperm Motility ; Spermatozoa

Ebola virus (EBOV) causes highly lethal hemorrhagic fever in humans and nonhuman primates and has a significant impact on public health. The nucleoprotein (NP) of EBOV (EBOV-NP) plays a central role in virus replication and has been used as a target molecule for disease diagnosis. In this study, we generated a monoclonal antibody (MAb) against EBOV-NP and mapped the epitope motif required for recognition by the MAb. The MAb generated via immunization of mice with prokaryotically expressed recombinant NP of the Zaire Ebola virus (ZEBOV-NP) was specific to ZEBOV-NP and able to recognize ZEBOV-NP expressed in prokaryotic and eukaryotic cells. The MAb cross-reacted with the NP of the Reston Ebola virus (REBOV), the Cote-d'Ivoire Ebola virus (CIEBOV) and the Bundibugyo Ebola virus (BEBOV) but not with the NP of the Sudan Ebola virus (SEBOV) or the Marburg virus (MARV). The minimal epitope sequence required for recognition by the MAb was the motif PPLESD, which is located between amino acid residues 583 and 588 at the C-terminus of ZEBOV-NP and well conserved among all 16 strains of ZEBOV, CIEBOV and BEBOV deposited in GenBank. The epitope motif is conserved in four out of five strains of REBOV.
Animals ; Antibodies, Monoclonal ; immunology ; Ebolavirus ; chemistry ; immunology ; Epitope Mapping ; methods ; Escherichia coli ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Nucleoproteins ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo perform pathological observation and etiological identification of specimens collected from dairy cows, beef cattle and dogs which were suspected of rabies in Inner Mongolia in 2011, and analyze their etiological characteristics.

METHODSPathological observation was conducted on the brain specimens of three infected animals with Hematoxylin-Eosin staining, followed by confirmation using immunofluorescence and nested RT-PCR methods. Finally, phylogenetic analysis was conducted using the virus N gene sequence amplified from three specimens.

RESULTSEosinophilic and cytoplasmic inclusion bodies were seen in neuronal cells of the CNS; and rabies non-characteristic histopathological changes were also detected in the CNS. The three brain specimens were detected positive. N gene nucleotide sequence of these three isolates showed distinct sequence identity, therefore they fell into different groups in the phylogenetic analysis. N gene in the cow and dog had higher homology with that in Hebei isolate, but that in the beef cattle had higher homology with that in Mongolian lupine isolate and Russian red fox isolate.

CONCLUSIONRabies were observed in the dairy cow, beef cattle and canine in the farm in Inner Mongolia, in 2011, which led to a different etiologic characteristics of the epidemic situation.

Acetazolamide ; Animals ; Brain ; pathology ; Cattle ; Cattle Diseases ; epidemiology ; pathology ; Dog Diseases ; diagnosis ; epidemiology ; Dogs ; Mongolia ; epidemiology ; Nucleoproteins ; genetics ; Phylogeny ; Rabies ; epidemiology ; veterinary ; Rabies virus ; genetics ; Time Factors