Genetic diversity and molecular evolution of Seoul virus in Hebei province, China.

Seoul virus (SEOV) is a major pathogen which causes hemorrhagic fever with renal syndrome (HFRS), and is present all over the world. However, there are currently few long-term systematic studies of SEOV's phylogenetic and evolutionary mechanisms in epidemic areas. Thus, in this study, we used RT-PCR combined with NGS to obtain the genomes of six SEOV viruses from 1993, as well as 56 Hebei province-specific tissue samples from 1999 to 2022. Phylogenetic analysis showed that the SEOV samples could be divided into seven groups and showed geographic clustering. The geographic region may be the main factor affecting the genetic diversity of SEOV. We also found that SEOV was subject to strong overall purifying selection and positive selection at certain sites during evolution. Recombination events and high nucleotide substitution rates were also shown to accelerate SEOV's evolution. Evolutionary feature of the L segment is more representative of complete genome. Our detailed analysis provides a deeper understanding of the genetic diversity and evolutionary drivers of SEOV within its primary epidemic areas. It will be important to further monitor epidemiological trends and drivers of variation to help increase our understanding of the pathogenicity of SEOV infections.

Atroposelective Access to Dihydropyridinones with C-N Axial and Point Chirality via NHC-Catalyzed [3 + 3] Annulation.

An N-heterocyclic carbene-catalyzed atroposelective [3 + 3] annulation of enals with 2-aminomaleate derivatives is described. A series of substituted dihydropyridones bearing both C-N axis and point chirality were synthesized with good diastereo- and enantioselectivity under mild conditions. This efficient strategy successfully superpositions an extra point chiral element with an axial backbone, and the generated structurally interesting atropisomers may have potential application in drug discovery.

Correction: Genetic variability of human adenovirus type 7 circulating in mainland China.

[This corrects the article DOI: 10.1371/journal.pone.0232092.].

Genetic variability of human adenovirus type 7 circulating in mainland China.

Human adenovirus (HAdV-7) is a highly contagious pathogen that causes severe respiratory illnesses. However, the epidemic patterns and genetic variability of HAdV-7 circulating in mainland China have not been well elucidated. In this study, we used Chinese HAdV sentinel surveillance data obtained from 2012-2015 to investigate the clinical features of 122 HAdV-7-positive cases and performed amplification and sequence determination of three capsid genes (penton base, hexon, and fiber) from 69 isolated viruses covering from seven provinces of China. Additionally, we compared with data from representative sequences of 21 strains covering seven more provinces in China and 32 international HAdV-7 strains obtained from GenBank database to determine the phylogenetic, sequence variations, and molecular evolution of HAdV-7. The results indicated that HAdV-7 infection occurred throughout the year, and a high proportion of severe cases (27 cases, 22.1%) exhibited infantile pneumonia. Moreover, phylogenetic analysis showed that all HAdV-7 strains could be divided into two major evolutionary branches, including subtype 1 and subtype 2, and subtype 3 was also formed according to analysis of the penton base gene. Subtypes 1 and 2 co-circulated in China before 2008, and HAdV-7 strains currently circulating in China belonged to subtype 2, which was also the predominant strain circulating worldwide in recent years. Further sequence variation analysis indicated that three genes of HAdV-7 were relatively stable across time and geographic space, particularly for viruses within subtypes, which shared almost the same variation sites. Owing to continuous outbreaks caused by HAdV-7, resulting in increased illness severity and fatality rates in China, the establishment of a national HAdV surveillance system is urgently needed for the development of effective preventive and infection-control interventions for adenovirus respiratory infections in China.

Long-term retrospective observation reveals stabilities and variations of hantavirus infection in Hebei, China.

BACKGROUND: Hemorrhagic fever with renal syndrome (HFRS) is an emerging zoonotic infectious disease caused by hantaviruses which circulate worldwide. So far, it was still considered as one of serious public health problems in China. The present study aimed to reveal the stabilities and variations of hantavirus infection in Hebei province located in North China through a long-term retrospective observation. METHODS: The epidemiological data of HFRS cases from all 11 cities of Hebei province since 1981 through 2016 were collected and descriptively analyzed. The rodent densities, species compositions and virus-carrying rates of different regions were collected from six separated rodent surveillance points which set up since 2007. The molecular diversity and phylogenetic relationship of hantaviruses circulating among rodents were analyzed based on partial viral glycoprotein gene. RESULTS: HFRS cases have been reported every year in Hebei province, since the first local case was identified in 1981. The epidemic history can be artificially divided into three phases and a total of 55,507 HFRS cases with 374 deaths were reported during 1981-2016. The gender and occupational factors of susceptible population were invarible throughout, however age of that was gradually aging. The annual outbreak peak always present in spring, while the main epidemic region had gradully altered from south to northeast. Surveillance of rodents revealed that residential rodents significantly possessed higher density and virus-carring rate than field rodents. The house rat, Rattus norvegicus, was the dominant rodent species and Seoul virus S3 sub-genotype which is continued but slightly evolving perhaps to be the sole pathogen for local HFRS cases of Hebei province. CONCLUSIONS: This long-term province-wide surveillance and epidemiological analysis has revealed the stabilities and variations of hantavirus infection in North China. In order to improve current prevention and control strategies of HFRS in China, all surveillance should be continuously enhanced and variations should be paid more attentions.

Seroprevalence, cross antigenicity and circulation sphere of bat-borne hantaviruses revealed by serological and antigenic analyses.

Bats are newly identified reservoirs of hantaviruses (HVs) among which very divergent HVs have been discovered in recent years. However, their significance for public health remains unclear since their seroprevalence as well as antigenic relationship with human-infecting HVs have not been investigated. In the present study archived tissues of 1,419 bats of 22 species from 6 families collected in 5 south and southwest provinces in China were screened by pan-HV RT-PCR following viral metagenomic analysis. As a result nine HVs have been identified in two bat species in two provinces and phylogenetically classified into two species, Laibin virus (LAIV, ICTV approved species, 1 strain) and Xuan son virus (XSV, proposed species, 8 strains). Additionally, 709 serum samples of these bats were also analyzed by ELISA to investigate the seroprevalence and cross-reactivity between different HVs using expressed recombinant nucleocapsid proteins (rNPs) of LAIV, XSV and Seoul virus (SEOV). The cross-reactivity of some bat sera were further confirmed by western blot (WB) using three rNPs followed by fluorescent antibody virus neutralization test (FAVNT) against live SEOV. Results showed that the total HV seropositive rate of bat sera was 18.5% (131/709) with many cross reacting with two or all three rNPs and several able to neutralize SEOV. WB analysis using the three rNPs and their specific hyperimmune sera demonstrated cross-reactivity between XSV/SEOV and LAIV/XSV, but not LAIV/SEOV, indicating that XSV is antigenically closer to human-infecting HVs. In addition a study of the distribution of the viruses identified an area covering the region between Chinese Guangxi and North Vietnam, in which XSV and LAIV circulate within different bat colonies with a high seroprevalence. A circulation sphere of bat-borne HVs has therefore been proposed.

Spatiotemporal patterns of hemorrhagic fever with renal syndrome in Hebei Province, China, 2001-2016.

Hemorrhagic fever with renal syndrome (HFRS) is highly endemic in China, where approximately 90% of the total human cases in the world are reported. The Hebei province, one of areas with the highest prevalence, has reported HFRS cases every year in the last two decades. This study describes the spatiotemporal patterns of HFRS in the Hebei province from 2001 to 2016, detects the high-risk spatiotemporal clusters of HFRS, and provides valuable information for planning and implementation of local preventive measures. For the purpose of the analysis, HFRS cases recorded during the sixteen years in the Hebei province were aggregated into three temporal periods (2001-2006, 2007-2012, and 2013-2016). Spatiotemporal analyses, including Global spatial autocorrelation analysis and Kulldorff's scan statistical analysis, were applied to analyze te spatiotemporal clusters of HFRS at the county level. The results revealed that the spatial extent of the HFRS epidemic in the Hebei province changed dynamically from 2001 to 2016, which indicated that a comprehensive preventative strategy should be implemented in the northeastern regions of the Hebei province in spring.

Genovariation Study of Hantavirus in Main Endemic Areas of Hemorrhagic Fever with Renal Syndrome in Hebei Province, China.

BACKGROUND: Hemorrhagic fever with renal syndrome (HFRS) is an important infectious disease in Hebei Province. At present, cases from the northeast regions of the province account for >80% of the total incidences. However, studies that examine the region-specific genetic variations of the Hantavirus (HV), the causative pathogen for HFRS, have been lacking. METHODS: Rodents were collected in northeast Hebei Province from 2004 to 2013, and the HV strains used in this study were isolated in 1993. Lung tissues were isolated from the rodents and HV antigen was detected by indirect immunofluorescence. The M1 and M2 fragments of HV M region were amplified by reverse transcription polymerase chain reaction (RT-PCR), cloned into pMDl9-T vector, sequenced and compared with representative standard stains for homology and phylogenetic analysis. RESULT: A total of 21 samples of HV antigen-positive were collected. Real-time PCR analysis revealed that the 19 rodent lungs and two HV strains were positive for the SEO virus. 11 samples were chosen to sequence, and they shared 95.8%-99.8% in nucleotide homology, and 83.6%-99.2% when compared to the standard strains of SEO virus. Phylogenetic analysis demonstrated that all strains were grouped into the same S3 subtype. CONCLUSION: SEO was the major epidemic genotype of HV in the main HFRS endemic areas in Hebei Province, and S3 was the major subtype. There was minor genetic variation in HV over short term periods, while long term variations were higher.

Analysis of incidence and related factors of hemorrhagic fever with renal syndrome in Hebei Province, China.

BACKGROUND: Hemorrhagic fever with renal syndrome (HFRS) is an important infectious disease in Hebei province, China. The average annual incidence is 3.10/100000. Although the incidence of HFRS is stable with a general decline in trend since 2004, an increase in the trend was observed in 2011. Few studies have been conducted to investigate the underlying risk factors for this disease. METHODS: The epidemiological data of HFRS and the population data, meteorology, and remote sensing aspects of cities in Hebei province from 1999 to 2011 were collected. The epidemiological data included the population density in the region, seasonal data and rat density and rat's virus carriage rate. The data were analyzed by descriptive study, correlation analysis and multivariate linear analysis. RESULTS: There were 26837 cases of HFRS was reported from 1999 to 2011. The infection occurred mainly in males, aged 20∼49 years, who were farmers. Rattus norvegicus was the main host animal. The incidence of HFRS was related to NDVI value, rat density and rat's virus carriage rate by multiple linear regression (F = 25.936, P<0.01). The pseudo- R2value for the model was 0.644. CONCLUSION: The incidence of HFRS was related to NDVI value, rat density and rat's virus carriage rate. Control of these factors should be used to prevent HFRS in Hebei province.

Application of an autoregressive integrated moving average model for predicting the incidence of hemorrhagic fever with renal syndrome.

The Box-Jenkins approach was used to fit an autoregressive integrated moving average (ARIMA) model to the incidence of hemorrhagic fever with renal Syndrome (HFRS) in China during 1986-2009. The ARIMA (0, 1, 1) × (2, 1, 0)(12) models fitted exactly with the number of cases during January 1986-December 2009. The fitted model was then used to predict HFRS incidence during 2010, and the number of cases during January-December 2010 fell within the model's confidence interval for the predicted number of cases in 2010. This finding suggests that the ARIMA model fits the fluctuations in HFRS frequency and it can be used for future forecasting when applied to HFRS prevention and control.

[Genotyping and sequence analysis on G2 segment of hantavirus carried by rat in Hebei province].

OBJECTIVE: To know the genotype and subtype of hantavirus (HV) carried by host animals in HFRS natural epidemic area of Hebei province. METHODS: According to the conservative sequence of 76-118 and R22 strains, specific primers were designed. RT-nested PCR was used to amplify partial M segments from the positive rat lungs screened by IFA. Agarose gel electrophoresis was used to identify the types. Nucleotides were assayed from partial products after purification and reclaim. Gene analysis was carried on with DNAStar package. RESULTS: 32 specimens, which were positive screened by IFA, were amplified the specific segment (418bp) and all belonged to type SEO. Sequencing results of 10 partial segments indicated that G2 segment had little variability and nucleotide homology reached to 98.0%-100.0%. Comparing with the R22 and 76-118 strains, homology was 93.3%-94.3% and 67.7%-69.0% respectively. CONCLUSION: According to G2 segment, SEO was the major type in Hebei HFRS natural epidemic area and S3 was the major subtype. HV which belonged to the same subtype had high homology and genetic materials were correspondingly stable. Different rats could carry the same subtype of HV.

[Genotype and sequence analysis on G2 segments of hantavirus from HFRS patients in Hebei Province].

OBJECTIVE: To know the genotype and subtype of hantavirus (HV) which infected persons in Hebei province. METHODS: According to G2 coding region of 76-118 and R22 strains, specific type primers were designed to detect and identity the types of HV in HFRS patients' sera with RT-nested PCR. Nucleotides were assayed from partial products after purification and reclaim. Then, gene analysis was done with DNAStar package. RESULTS: 17 out of 69 positive serum specimens were successfully detected by RT-PCR and the detection rate was 24.64%, among which, or= 14 days were 0. 17 positive specimens were all belonged to SEO. The nucleotide homology of 9 typical specimens was 92.0%-100%. Between HeB7 and other 8 specimens was 92%-95%, and they belonged to different subtypes. When HeB7 compared with R22 strain, it was 97.7%. HeB7 and R22 belonged to S1 subtype. The 8 specimens except HeB7 was 95.7%-100% and they all belonged to S3 subtype. When compared with 76-118 strain, 9 specimens' nucleotide homology was only 70.3%-72.7%, belonged to different type. CONCLUSION: SEO was the major type of HV from HFRS patients in Hebei province, S3 was the major subtype and S1 was also existed. In a certain area, the HV which belonged to the same type was correspondingly conservative, and had the characteristic of regional stability.