The prenatal weekly temperature exposure and neonatal congenital heart disease: a large population-based observational study in China.

We aim to explore the link between maternal weekly temperature exposure and CHD in offspring and identify the relative contributions from heat and cold and from moderate and extreme atmospheric temperature. From January 2019 to December 2020, newborns who were diagnosed with CHD by echocardiography in the Network Platform for Congenital Heart Disease (NPCHD) from 11 cities in eastern China were enrolled in the present study. We appraised the exposure lag response relationship between temperature and CHDs in the distributed lag nonlinear model and further probed the pooled estimates by multivariate meta-analysis. We further performed the exposure-response curves in extreme temperature (5th percentile for cold and 95th for hot events). We also delve into the cumulative risk ratios (CRRs) of temperature on CHDs in general and subgroups. In this study, 5904 of 983, 523 infants were diagnosed with CHDs. The temperature-CHD combination performed positive significance in two exposure windows, gestational weeks 10-16 and 26-31, and reached the maximum effect in the 28th week. Compared with extreme cold (5th, 6.14℃), these effects were higher in extreme heat (95th, 29.26℃). The cumulative exposure-response curve showed a steep nonlinear rise in the hot tail but showed non-significance at low temperatures. In this range, the CRRs of temperature showed an increment to a ceiling of 3.781 (95% CI: 1.460-10.723). The temperature- CHD curves for both sex groups showed a general growth trend. No statistical significance was observed between these two groups (P = 0.106). The cumulative effect of the temperature related CHD was significant in regions with lower education levels (maximum CRR was 9.282 (3.019-28.535)). A degree centigrade increase in temperature exposure was associated with the increment of CHD risk in the first and second trimesters, especially in extreme heat. Neonates born in lower education regions were more vulnerable to temperature-related CHDs.

Geographical and Socioeconomic Factors Influence the Birth Prevalence of Congenital Heart Disease: A Population-based Cross-sectional Study in Eastern China.

Neonatal congenital heart disease (CHD) is the most common congenital anomaly. As a practical matter of people's livelihood, cardiac ultrasonography was performed on potential CHD children in 11 cities eastern China. In this study, we aimed to document the birth prevalence of CHD and its socioeconomic and geographical distribution, as supported by this public health policy. In this study, the diagnosis of CHD was made based on echocardiography. Geographical and socioeconomic factors were determined by the Statistical Bulletin on National Economic and Social Development (SBNESD). 51857 newborns from the Network Platform for Congenital Heart Disease (NPCHD) from January to December 2019 in 11 cities eastern China were included. The total birth prevalence of CHD was 5.79 per 1000 births. The study on the low-income areas, mountainous areas, areas with low medical institution bed level, and with high qualification of medical personnel reported a signifcantly higher birth prevalence of CHD compared with high-income cities, flat areas, areas with high medical institution bed level, and with low qualification of medical personnel. ASD, VSD, PDA, PS, TOF, atrioventricular septal defect, coarctation of the aorta, TAPVD, TGA and pulmonary atresia are the most frequent subtypes. ASD, VSD, PDA, PS, atrioventricular septal defect, coarctation of the aorta and pulmonary atresia showed a female preponderance, while TOF, TGA and TAPVD showed a male preponderance. Our study gives a relatively realistic prevalence of CHD after cardiac ultrasound examination of newborns suspected positive with CHD. Significant differences across geographical regions, income levels, and health service access were observed. In the future, population-wide cardiac ultrasound screening, prospective birth defect registries, and systematic medical follow-up programs covering the entire eastern or even China are needed to determine the exact birth prevalence.

Maternal air pollution exposure and neonatal congenital heart disease: A multi-city cross-sectional study in eastern China.

OBJECTIVE: To examine the association between air pollution and neonatal congenital heart disease (CHD), and evaluate the cumulative burden of CHD attributed to above certain level for ambient air pollution exposure. METHODS: We identified newborns who were diagnosed as CHD by echocardiography in Network Platform for Congenital Heart Disease (NPCHD) from January 2019 to December 2020 in 11 cities eastern China. The exposure lag response relationship between air pollutants (PM2.5, PM10, SO2, NO2, CO, and O3) concentration and CHDs was calculated by the distributed lag nonlinear model (DLNM). We further calculated the cumulative risk ratios (CRRs) of each air pollutant above reference concentrations on CHDs. RESULTS: A total of 5904 CHDs from 983, 523 newborns were enrolled in this study. A 10 µg/m3 increase in PM2.5, PM10, SO2, NO2, CO and O3 exposure was associated with an increased risk of higher CHD incident RR = 1.025, 95% CI: 1.016-1.038 for PM2.5 in the third trimester, RR = 1.001, 95% CI: 1.000-1.002 for PM10 in the third trimester, 1.020, 95%CI: 1.004-1.036 for NO2 in the third trimester, RR = 1.001, 95%CI: 1.000-1.002 for O3 in the first trimester, all P value < 0.05). Cumulative effect curves of PM2.5, PM10, SO2, NO2, CO, and O3 were observed as sub-linear with a maximum of 1.876 (95%CI:1.220-2.886), 1.973 (95%CI:1.477,2.637), 2.169 (95%CI:1.347-3.493), 2.902 (95%CI:1.859-4.530), 1.398 (95%CI:1.080-1.809), 2.691 (95%CI:1.705-4.248), respectively. Significant associations were observed for air pollutants and CHDs in cities with higher average education years and babies concepted in cold season. CONCLUSIONS: Our findings could provide growing evidence regarding the adverse health effects of air pollution on CHD, thereby strengthening the hypothesis that air pollutants have harmful impacts on cardiac development. Further studies are needed to verify the associations.

Case Report: Prenatal Diagnosis of a Fetus With Harlequin Ichthyosis Identifies Novel Compound Heterozygous Variants: A Case Report.

BACKGROUND: Harlequin ichthyosis (HI) is the most severe form of the keratinizing disorders, and it is characterized by whole-body hard stratum corneum. ABCA12 has been identified as the major disease-causing gene of HI. METHODS: A case of HI was prenatally diagnosed by ultrasonography and genetic tests. The fetus had been found with dentofacial deformity and profound thickening of the palm and plantar soft tissues. Chromosomal microarray analysis (CMA) and whole exome sequencing (WES) were then performed on the amniotic fluid to identify germline pathogenic variants for the fetus. Candidate variants were verified by Sanger sequencing. RESULTS: Compound heterozygous frameshift variants (p.Q719QfsX21; p.F2286LfsX6) of ABCA12 were identified for the fetus, suggesting the former variants were maternally inherited and the latter paternally inherited. The fetus was terminated. CONCLUSION: A prenatal molecular diagnosis is an important approach for the prevention of HI. In the study, we provided a successful case of genetic counseling for a family with an HI baby.

Paternal Low-Level Mosaicism-Caused SATB2-Associated Syndrome.

Mutations of SATB2 (OMIM#608148) gene at 2q33.1 have been associated with the autosomal dominant SATB2-associated syndrome (SAS), which is still short of comprehensive diagnosis technologies for small deletions and low-level mosaicism. In this Chinese Han family, single nucleotide polymorphism array identified a 4.9-kb deletion in the SATB2 gene in two consecutive siblings exhibiting obvious developmental delay and dental abnormalities but failed to find so in their parents. Prenatal diagnosis revealed that their third child carried the same deletion in SATB2 and the pregnancy was terminated. To determine the genetic causes behind the inheritance of SATB2 deletion, gap-PCR was performed on peripheral blood-derived genomic DNA of the family and semen-derived DNA from the father. Gap-PCR that revealed the deletions in the two affected siblings were inherited from the father, while the less intense mutant band indicated the mosaicism of this mutation in the father. The deletion was 3,013 bp in size, spanning from chr2: 200,191,313-200,194,324 (hg19), and covering the entire exon 9 and part of intron 8 and 9 sequences. Droplet digital PCR demonstrated mosaicism percentage of 13.2% and 16.7% in peripheral blood-derived genomic DNA and semen-derived DNA of the father, respectively. Hereby, we describe a family of special AT-rich sequence-binding protein 2-associated syndrome caused by paternal low-level mosaicism and provide effective diagnostic technologies for intragenic deletions.

[Phenotypic and genetic analysis of a sibpair with partial deletion of SATB2 gene caused by 2q33.1 microdeletion].

OBJECTIVE: To analyze the genotype and phenotype of a sibpair with partial deletion of SATB2 gene caused by 2q33.1 microdeletion. METHODS: Both children have featured mental retardation and development delay, and were subjected to karyotyping, single nucleotide microarray (SNP array) and real-time fluorescence quantitative PCR analysis. Karyotyping and SNP Array analysis were also carried out on their parents to verify the origin of mutation. RESULTS: Both sibs had a normal karyotype. SNP array showed that sib 1 had arr[hg19]2q33.1(200 192 328 - 200 197 269)×1 (4.9 kb), 2q35 (218 105 663 - 218 816 675)×3 (711 kb), while sib 2 had arr[hg19]2q33.1(200 192 328 - 200 197 269)×1 (4.9 kb), 2q35 (218 105 663-218 810 908)×3 (705.2 kb). The deletion has partially overlapped with that of 2q33.1 microdeletion syndrome and involved part of the SATB2 gene. The result of real-time fluorescence quantitative PCR assay was consistent with that of SNP assay. The duplication has originated from their father and has not been associated with any disease phenotypen. CONCLUSION: Both sibs have carried partial deletion of SATB2 gene and had similar clinical phenotypes. Haploinsufficiency of such gene probably underlies the clinical manifestations in both patients.

A multiplex liquid-chip assay based on Luminex xMAP technology for simultaneous detection of six common respiratory viruses.

We utilized one-step multiplex reverse transcription-PCR (RT-PCR) and Luminex xMAP technology to develop a respiratory multiplex liquid-chip assay (rMLA) for simultaneous detection of 6 common respiratory viruses, including influenza virus type A (FluA) and type B (FluB), para-influenza virus type 3 (PIV-3), respiratory syncytial virus (RSV), human metapneumovirus (MPV) and a threatening virus to China, Middle East Respiratory Syndrome coronavirus (MERS-CoV). Performance of rMLA was evaluated by comparing with real-time RT-PCR. Detection data from clinical specimens showed that the rMLA had diagnostic sensitivities of 97.10% for FluA, 94.59% for FluB, 98.68% for PIV-3, 94.87% for RSV and 95.92% for MPV (No Data for MERS-CoV due to the lack of positive specimens). Data of analytical sensitivities showed that the detection limits of the rMLA assay were 5-25 viral RNA copies per µl for FluA, FluB, PIV-3 and MERS-CoV, approximate to the real-time RT-PCR assay; while the values were 8 and 22copies/µl for MPV and RSV, lower than the real-time RT-PCR(78 and 114 copies/µl respectively). The results indicated that the rMLA is a sensitive, specific detection tool and comparable to real-time RT-PCR, especially suitable for high-throughput detection of respiratory specimens.

Investigation of rare chronic lipoid pneumonia associated with occupational exposure to paraffin aerosol.

OBJECTIVES: Occupational exposure to paraffin is an infrequent cause of lipoid pneumonia (LP) and related data are scare. We investigated the possible relationship between three rare cases of chronic LP and occupational exposure to paraffin aerosol in an iron foundry. METHODS: The three cases of LP and their workplaces were investigated using data from field investigations, air monitoring, pulmonary radiological examinations, cell staining, and lung biopsies. RESULTS: The patients had long-term occupational exposure to paraffin. X-ray diffraction testing revealed that the raw material from the workshop was paraffin crystal. The air concentrations of paraffin aerosol in workplaces were significantly higher than outdoor background levels. Small diffuse and miliary shadows with unclear edges were observed throughout the whole lungs via radiography. Computed tomography revealed diffuse punctate nodules and a high density of stripe-like shadows in both lungs (ground-glass opacity in a lower lobe, and a mass-like lesion and high translucent area near the bottom of the lung). Lipid-laden macrophages were found in the sputum and bronchial lavage. A broadened alveolar septum and local focal fibrosis were also discovered via lung biopsy. The inflammatory reaction in the lung tissues appeared to resolve over time. CONCLUSIONS: These three rare cases of chronic LP in workers during molding and repair processes were associated with occupational paraffin aerosol exposure. Therefore, primary prevention is essential for molding or repairing workers in the iron foundry, and a differential diagnosis of occupational chronic LP (vs. pneumoconiosis) should be considered when treating these workers.

Macrophage infiltration promotes invasiveness of breast cancer cells via activating long non-coding RNA UCA1.

There is now considerable evidence supporting the view that macrophage infiltration is playing a critical role in the proliferation and progression of breast cancer but the underlying molecular mechanisms remain largely unknown. To this end, using long non-coding RNA (lncRNA) expression profiling, we examined changes in lncRNA expression in breast cancer cells treated with conditioned medium (CM) from cultured human THP-1 macrophages. We found that treatment with macrophage CM induced the expression of numerous lncRNAs, including urothelial cancer associated 1 (UCA1). Knockdown of UCA1 using shRNA inhibited AKT phosphorylation and abolished invasiveness of tumor cells induced by macrophage CM. Consistent with these results; we further showed that UCA1 level was significantly enhanced in human primary breast tumors and correlated with advanced clinical stage, supporting its role in promoting carcinogenesis and progression of breast cancer. Together, these results suggest that macrophage could promote invasiveness of breast cancer cells by enhancing expression of lncRNA UCA1.

[Sequencing and analysis of N gene of street rabies virus isolated from different hosts in Zhejiang province].

OBJECTIVE: To elucidate the characteristics of genetic variability and its relationship with prevalence, through sequencing and analysis of N gene among street rabies virus isolated from different hosts (homo sapiens, ferret badger, dog) in Zhejiang province. METHODS: Samples were screened and confirmed by direct fluorescence assay and reverse transcript PCR. Sequences were analyzed using bio-information software. RESULTS: Eighteen street rabies virus strains were identified, including 2 from homo sapiens, 5 from ferret badger, and 11 from dog. Similarities of N gene and N protein were calculated to be 89.7%-100.0% and 98.4%-100.0% respectively. Mutations occurred in N gene were almost non-sense mutations. In addition,Data from phylogenetic analysis showed that all these strains could be classified into traditional genotype 1. CONCLUSION: The prevalence of rabies viruses among different hosts in Zhejiang province had certain regional properties. Rabies viruses isolated from the same kind of host or from the same/adjacent county/counties had the closest relationship. However, the characteristics of rabies virus prevalent in homo sapiens were somewhat complicated. In summary, the transmission of street rabies virus in Zhejiang province was from dogs to ferret badgers and homo sapiens, and the virus could circulate and cross-regional transmit among dogs and ferret badgers.

[Tracing investigation and diagnosis of one transfusion-transmitted malaria case in Zhejiang Province].

OBJECTIVE: To identify the sources of infection and the mode of transmission of a malaria case with unknown origin. METHODS: Clinical data of the case were collected and the epidemiological investigation was conducted. The blood samples of the patient and the suspected infection source (blood donor) were detected by microscopy, rapid diagnostic test strip (RDT) and nested PCR. RESULTS: The patient did not visited malaria endemic areas. After a blood transfusion, the patient had chills and fever, and was confirmed as falciparum malaria by microscopy with bone marrow and peripheral blood smears and RDT. The blood donor was a worker returned from Africa. Before blood donation she was sick like malaria carrier, and took anti-malarial drug. She was then confirmed as falciparum malaria by RDT and microscopy. The blood samples from the patient and the blood donor were diagnosed as falciparum malaria by nested PCR, and the similarity of the small subunit rRNA (SSU rRNA) sequence was 100%, showing they were mix-infected with K1 and MAD20 genotypes of Plasmodium falciparum. CONCLUSION: This patient is confirmed P. falciparum infection via blood transfusion from a donor who returned from Africa.

[Secretory expression of salivary ATP diphosphohydrolase (apyrase) from Aedes albopictus in Pichia pastoris].

The gene-coding mature apyrase protein from Aedes albopictus was amplified by RT-PCR and cloned in frame with the a-factor secretion signal peptide into Pichia pastoris secreting expression vector pGAPZalpha-A resulting in the pGAPZa-A-apyrase. After being linearized by Bln I restriction enzyme, the recombinant pGAPZalpha-A-apyrase was trans-formed into Pichia pastoris GS115 by electroporation. Recombinant strains pGAPZalpha-A-apyrase/GS115 were screened on YPDS plates containing Zeocin and identified by PCR. The recombinant protein of apyrase (M(r) 60000) has been expressed in the supernatant of Pichia pastoris.

[Laboratory testing for a case of imported Plasmodium ovale infection in Zhejiang Province].

Blood sample obtained from a patient, which returned from Equatorial Guinea, with clinical diagnosis of Plasmodium infection was confirmed as imported P. ovale infection by etiology and molecular biological methods. 50 microl blood was obtained before taking anti-malarial drugs to make thin and thick blood smears, Giemsa stained, and observed by microscopy. Genomic DNA was extracted from the blood sample, and detected for DNA fragment of P. ovale, P. vivax, P. falciparum or P. malariae by real-time fluorescent quantitative PCR. P. ovale parasites were found in both thin and thick blood smears, and confirmed by quantitative PCR. With the results of laboratory testing, epidemiological history and clinical manifestations, the patient was diagnosed as imported P. ovale infection.

[Pathogen identification and clinical diagnosis for one case infected with Babesia].

OBJECTIVE: To identify the pathogen and make diagnosis on a case who was misdiagnosed as malaria. METHODS: Clinical and epidemiological information of the suspected case was collected. Blood samples during hospitalization were collected and examined microscopically. Genomic DNA from the blood samples was amplified by Babesia 18S RNA genus- and species- specific primers, respectively, and the amplified products were used in sequencing and BLAST sequence analysis. RESULTS: The case had a fever over 20 days repeatedly with anaemia (RBC 2.59 x 10(12), HB 5.5 g/L) and hepatosplenomegaly. The unidentified parasites were found in the bone marrow and blood smear after Giemsa staining. Epidemiological information revealed that this case had a history of blood transfusion and tick bites. 1 625 bp and 449 bp band generated by PCR amplification from blood sample using Babesia genus- and species-specific primers, and the sequence homology was 99% in comparison to Babesia microti (AB241632) with BLAST analysis. CONCLUSION: The clinical information, epidemiological history, and the PCR identification confirm the diagnosis of Babesia microti infection.

[Temporal and spatial population dynamics of rabies virus isolates in China].

In order to study phylogeography, population dynamics and molecular evolution of rabies viruses (RABVs) isolates from China, especially spatio-temporal dynamics, the timescale of RABVs evolution and its pattern of migration, we performed an extensive comparative analysis of RABV N gene sequence data, representing 167 isolates sampled from 20 provinces in a 78-year period (from 1931 through 2009). The available Chinese isolates could be divided into two distinct clades:Phylogroup clades I comprised Chinese group 1-4; Phylogroup clades II contained Chinese group 5-8. We found no evidence for positive selection (dN/dS>1) acting at any codon and found strong selective constraints for N gene. Bayesian Markov Chain Monte Carlo (MCMC) analysis suggested that the Chinese rabies viruses originated within the last 2000 years and the mean rates of nucleotide substitution for the N gene were approximately 4 x 10(-4) substitutions per site per year. The analyses of the spatial and spatio-temporal evolution indicated that RABV isolates from China migrated among different Provinces.

Evolutionary dynamics of rabies viruses highlights the importance of China rabies transmission in Asia.

Rabies in Asia is emerging as a serious public health issue. To explore the possible origin, phylogenetic relationships, and evolutionary dynamics of Asian Rabies viruses (RABV), we examined 200 complete nucleoprotein (N) gene sequences from RABV isolates in the region. Phylogeny supported the classification of Asian RABVs into five distinct clusters in lyssavirus genotype 1. Our geospatial and temporal analyses demonstrated that China appears to be the prime source of Asian RABVs. Understanding of rabies transmission and associated human activities, such as dog translocation, can help rabies control and elimination in Asia through collaborative efforts or programs.

[Molecular epidemiology investigation of rabies virus nucleoprotein genes in China].

OBJECTIVE: To study the relationship between the distribution of rabies virus and genetic variation, the genetic characterization and variation of rabies virus strains in China were analyzed. METHODS: The downstream 720 nucleotides of Nucleoprotein (N) gene coding region of the rabies specimens from different areas and host animals were sequenced, and then homology and phylogenesis were analyzed. RESULTS: Nucleotide similarities of 34 N gene sequences were 87.5%-100%, and the deduced amino acid similarities were 93.3%-99.6%. Most of the nucleotide variations were synonymous mutations. CONCLUSION: The 34 rabies specimens all belong to genotype I and are of regional characteristic. The rabies viruses in high-incidence areas in China are of various origins and present the transmission tendency from high-incidence areas to surrounding regions. There may be cross-infection and mutual spread of rabies virus between wildlife and domestic animals as well as native and foreign animals.

Transmission dynamics of rabies in China over the last 40 years: 1969-2009.

BACKGROUND: Rabies is a serious reemerging zoonosis in China. The molecular evolution and transmission patterns of rabies virus inferred from historical data can provide guidelines for better disease control and prevention in the future. OBJECTIVES: To investigate the epidemiology and evolutionary dynamics of the rabies virus in China. STUDY DESIGN: The molecular evolution of 132 viral glycoprotein gene sequences of Chinese rabies viruses collected in 17 provinces and 3 municipalities between 1969 and 2009 was analyzed. RESULTS: Phylogenetic analysis revealed that Chinese rabies viruses are subdivided into 6 lineages (A-F) within Lyssavirus genotype 1. Lineage A represents the widely dispersed cosmopolitan lineage while lineage B is closely related to Arctic-like rabies viruses. The remaining lineages (C-F) are typical of those circulating across much of Southeast Asia. The evolutionary rate for Chinese rabies virus was 1.532 x 10(-4) substitutions per site per year, and the corresponding common ancestor was in about 1115. CONCLUSIONS: The phylogeographic structure demonstrated Chinese rabies viruses have been transmitted intra-provincially and extra-provincially due to human-related activities.

[Sequencing and analyses on glycoprotein gene of rabies viruses isolated in Zhejiang province, China].

OBJECTIVE: Based on sequencing the genomes of glycoprotein (GP) gene of rabies viruses isolated in Zhejiang, we analyzed the properties of rabies viruses genetic variation in molecular level, and to compare with those of other representative vaccine strains and street virus strains, get the information about rabies viruses variation. METHODS: Suckling mice against rabies virus were selected. Overlapped fragments were amplified by RT-PCR and full-length genomes were assembled to analyze the nucleotide and deduced protein similarities and phylogenetic analyses of the GP genes. RESULTS: The fourteen full-length genomes were completely sequenced and they had the same genetic structure with 1575 nts and deduced protein with 524 aa. Genetic analysis revealed that the nucleotide and amino acid homologies of GP gene from Zhejiang strains and other vaccine strains or street virus strains were 82.3% - 99.9% and 85.1% - 99.8%. The fourteen strains were genotype 1 according to the phylogenetic analyses. The GP amino acids of Zhejiang strains rabies virus strains without any recombination occurred in GP and no larger variation appeared in the major antigenic sites. CONCLUSION: The comprehensive analysis based on the first-level structure of GP demonstrated that it was possible that some advantageous antigenic epitopes existed in certain areas and potential antigenic determinants. It was evident that the GP gene of Zhejiang strains appear to be stable and their sequence similarity with the representative strains of street virus in China were higher than those of other vaccine strains. Some differences showed in the genetic structure and evolution relationship among Zhejiang strains, other street strains in other regions and vaccine strains.

[Sequencing and analysis of complete genome of rabies viruses isolated from Chinese Ferret-Badger and dog in Zhejiang province].

Based on sequencing the full-length genomes of four Chinese Ferret-Badger and dog, we analyze the properties of rabies viruses genetic variation in molecular level, get the information about rabies viruses prevalence and variation in Zhejiang, and enrich the genome database of rabies viruses street strains isolated from China. Rabies viruses in suckling mice were isolated, overlapped fragments were amplified by RT-PCR and full-length genomes were assembled to analyze the nucleotide and deduced protein similarities and phylogenetic analyses from Chinese Ferret-Badger, dog, sika deer, vole, used vaccine strain were determined. The four full-length genomes were sequenced completely and had the same genetic structure with the length of 11, 923 nts or 11, 925 nts including 58 nts-Leader, 1353 nts-NP, 894 nts-PP, 609 nts-MP, 1575 nts-GP, 6386 nts-LP, and 2, 5, 5 nts- intergenic regions(IGRs), 423 nts-Pseudogene-like sequence (psi), 70 nts-Trailer. The four full-length genomes were in accordance with the properties of Rhabdoviridae Lyssa virus by BLAST and multi-sequence alignment. The nucleotide and amino acid sequences among Chinese strains had the highest similarity, especially among animals of the same species. Of the four full-length genomes, the similarity in amino acid level was dramatically higher than that in nucleotide level, so the nucleotide mutations happened in these four genomes were most synonymous mutations. Compared with the reference rabies viruses, the lengths of the five protein coding regions had no change, no recombination, only with a few point mutations. It was evident that the five proteins appeared to be stable. The variation sites and types of the four genomes were similar to the reference vaccine or street strains. And the four strains were genotype 1 according to the multi-sequence and phylogenetic analyses, which possessed the distinct district characteristics of China. Therefore, these four rabies viruses are likely to be street viruses already existing in the natural world.