Objective To investigate the status on children of 3-14 years old who suffered from cerebral palsy in Liaoning province. Methods One thousand three hundred and twenty-three cases of children with cerebral palsy of 3-14 years old who received rehabilitation in city hospital, county hospital and community hospital were investigated from January 2013 to October 2016 in 14 cities in Liaoning Province. The proportion of cerebral palsy children in 3-4 years old, 4-5 years old, 8-9 years old, 5-6 years old , 6-7 years old and 7-8 years old was about 10%, and in the other age the proportion was about 7%. The proportion of men and women generally was 4:1;neonatal convulsion (252 cases, 19%), premature delivery (230 cases , 17.3%) and low birth weight infant (187 cases, 14.1%) were main risk factors and accounted for more than 10%. Spastic type cerebral palsy accounted for the highest proportion (54.35%, 719 cases)and ataxia cerebral palsy accounted for the lowest proportion (2.95%). In complications , lower intelligence accounted for the highest proportion (50.34%, 666 cases), followed by the language barrier (43.99% , 582 cases), and the other complications accounted for less than 10%.;gross motor function classification in most studied children was stageⅡ(35%) and stageⅢ(32.50%); 6.95% patients could go to school, and 84.96% patients had health insurance. Patients coming from city accounted for 69.01%, and patients coming from rural area accounted for 30.99%. Mothers′ education below primary school was 4.16% . 36.05% children received rehabilitation in comprehensive hospital, 60.09%in children′s hospital and 3.85%in maternal and child health hospital. Conclusions Spastic cerebral palsy is the main type of children with cerebral palsy in Liaoning.High risk factors include neonatal convulsions, premature birth and low birth weight infants. Most patients complicate with low intelligence and language barriers.This paper can be used as the basis of further research on prevention and treatment

OBJECTIVETo study the viremia formation in guinea-pigs infected with wild type and attenuated Japanese encephalitis virus (JEV).

METHODSGuniea pigs were inoculated intraperitoneally with different wild JEV strains and the attenuated vaccine strain and its parent virulent strain. Viremia was detected on different days following virus inoculation.

RESULTSAll the guinea-pigs inoculated with the wild JEV strains induced different levels of viremia (1.00-3.40 Lg pfu) on the 1st and 3rd day post inoculation. Using a virus titer of 10(4) pfu for inoculation, the animals inoculated with the SA14 parent strain induced relatively high viremia (10(2.4)-10(3.4) pfu), however no viremia coulds be detected on any tested days.

CONCLUSIONThe degree of viremia in guinea pigs can be used as a new method to evaluate the attenuation of JEV.

Animals ; Disease Models, Animal ; Encephalitis Virus, Japanese ; pathogenicity ; physiology ; Encephalitis, Japanese ; virology ; Guinea Pigs ; Humans ; Japanese Encephalitis Vaccines ; administration & dosage ; adverse effects ; Vaccines, Attenuated ; administration & dosage ; adverse effects ; Viremia ; virology ; Virulence ; Virus Replication

The biological and genetic characteristics of a highly neurovirulent JE virus strain SA4 were studied. Mice were inoculated intracerebrally with strain SA4 and SA14, and observed for 14 days, respectively. On different days, mice brains were harvested for titrations of the virus content in the brains. Full-length genome of SA4 was sequenced and compared with SA14 as well as other JE virus strains in the world. The results indicated that the mice inoculated by SA4 induced sickness and death more rapidly (24 hours faster) than those induced by the SA14. The virus titers in the brains of mice infected with SA4 were 0.5-1.0 lg PFU/mL higher than that infected with SA14. The sequence comparison indicated that the nucleotide and amino acid homology between SA4 and the other 21 JE strains were 84.6%-99.0% and 95.2%-99.7% respectively. Comparison with strain SA14 revealed that there were 17 amino acid differences between the two strains, of which 5 were in the E protein region. The results demonstrate that strain SA4 is a highly neurovirulent strain. The substitutions of the 17 amino acids in the SA4 strain can be the molecular basis for the biological characteristics of high neurovirulence.
Animals ; Brain ; virology ; Encephalitis Virus, Japanese ; classification ; genetics ; isolation & purification ; pathogenicity ; Encephalitis, Japanese ; mortality ; virology ; Genotype ; Humans ; Mice ; Sequence Analysis ; Viral Envelope Proteins ; genetics ; Virulence

OBJECTIVETo understand the molecular epidemiologic characteristics of hantavirus Shandong isolate JNL virus strain.

METHODSThe complete M and S gene of the JNL virus isolated from Shandong Province was amplified by RT- PCR, and the purified PCR product was cloned into T vector for sequencing.

RESULTSThe results revealed that the JNL M segment was 3615 bp in length, encoding 1135 amino acids, and the S segment was 1698 bp encoding 429 amino acids, JNL belongs to HTN virus. The comparison of homology with HTN and SEO types showed that the difference of M and S complete sequences between JNL and all other HTN virus strains reached 20.0%-20.6%, and 15.5%-16.0%, respectively. Phylogenetic tree also showed that the position of JNL is located at a different clade.

CONCLUSIONSHTN virus Shandong local isolate JNL strain is a new specific HTN subtype virus.

DNA, Viral ; analysis ; Hantaan virus ; classification ; genetics ; isolation & purification ; Hemorrhagic Fever with Renal Syndrome ; virology ; Humans ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Sequence Homology, Nucleic Acid

In order to reveal the phenotypic characteristics of 17 JE virus strains isolated from different years, plaque sizes, mice neurovirulence and mice neuroinvasiveness of the isolates were studied and compared. BHK21 cell monolayers were used for testing the plaque sizes. The virus neurovirulence was tested in 9-11g mice inoculated intracerebrally and the virus neuroinvasiveness was tested in 9-11g and 14-16g by subcutaneous inoculation. Results showed that all the viruses produced clear plaques on the BHK21 cell monolayers with different sizes and all the virus strains appeared high neurovirulence in the mice with higher than lg8. 0/0.03 mL virus titers, while no apparent difference among them. The neuroinvasiveness (subcutaneous virulence) tested in the 9-11g mice had shown a little difference, but when tested in the 12-14 g mice,the difference was apparent. The results demonstrated that JEV in nature were highly neurovirulent with no apparent difference. However the neuroinvasiveness of the JEV in nature was greatly different, which didn't relate to the years of isolation and genotypes, but most of the viruses isolated from patients showed higher neuroinvasiveness.
Animals ; Cell Line ; China ; Culicidae ; virology ; Encephalitis Virus, Japanese ; genetics ; isolation & purification ; pathogenicity ; Encephalitis, Japanese ; virology ; Genotype ; Humans ; Mice ; Phenotype ; Viral Plaque Assay ; Virulence

CTN-1 is one of the rabies vaccine strains for human use in China, but there has been no report on the full-length gene sequence of CTN-1. In this study, the full-length gene of CTN-1 was amplified by RT-PCR, each PCR product was cloned into T vector and then sequenced, assemblied and compared with other vaccine strains as well as the wild Chinese rabies isolates. The phylogenetic tree of G gene was constructed and the genetic homology was analyzed. The results revealed that CTN-1 was 11 925nt (GenBank accession number: FJ959397)in length and belonged to the genotype I. The full-length nucleotide homologies among CTN-1 and other rabies virus strains were between 81.5%-93.4%, of which the lowest 81.5% was between CTN-1 strain and bat isolate SHBRV, and the highest 93.4% was between CTN-1 and Chinese isolate HN10. The phylogenetic analysis revealed that the majority of Chinese isolates could be grouped into the same clade with the CTN-1 strain, but aG and some vaccine strains from abroad such as Flury, PM, PV, ERA, RC-HL and a few Chinese strains were grouped in another clade. Comparsion of the G protein genes also showed that the homologies among CTN-1 and most of the Chinese isolates were higher than that of the other vaccine strains to those Chinese strains. Therefore, it suggests that the CTN-1 strain is more suitable and rational to be used for the production of rabies inactivated vaccine in China than the others.
Genome, Viral ; genetics ; Humans ; Molecular Sequence Data ; Phylogeny ; Rabies ; prevention & control ; virology ; Rabies virus ; classification ; genetics ; immunology ; Sequence Alignment ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; Viral Vaccines ; genetics

OBJECTIVETo construct the anti-lung tumor gene differentially expressed bank of wild mouse and to explore the mechanisms of the TW wild mouse suppressing the occurring of lung tumor.

METHODSUsing suppression subtractive hybridization (SSH) technique, the differentially expressed genes between TAF1 mouse and A/wy mouse were selected out and the subtracted cDNA bank was constructed. 166 clones were performed DNA sequencing and then were assayed by blast programme.

RESULTSAmong the blast results of 166 differentially expressed clones, 87 known genes (mRNA or cDNA) were in homology with 134 clones and were divided into 7 classifications according to the biological role.14 DNA fragments were in homology with 32 clones, in which 20 clones were in homology with 9 mouse DNA sequences, 2 clones were in homology with one bacterial gene sequences and 3 clones were clone vector.

CONCLUSIONWith SSH technique, the anti-lung tumor gene differentially expressed bank of wild mouse are successfully constructed.

Animals ; DNA, Complementary ; genetics ; Female ; Gene Expression Profiling ; Gene Library ; Lung Neoplasms ; genetics ; Male ; Mice ; Mice, Inbred Strains ; genetics ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; genetics ; Tumor Cells, Cultured

To sequence and analyze the full-length gene sequence of rabies vaccine virus aG strain. The full-length gene sequence of aG strain was amplified by RT-PCR by 8 fragments,each PCR product was cloned into vector pGEM-T respectively, sequenced and assemblied; The 5' leader sequence was sequenced with method of 5' RACE. The homology between aG and other rabies vaccine virus was analyzed by using DNAstar and Mega4. 0 software. aG strain was 11 925nt(GenBank accession number: JN234411) in length and belonged to the genotype I . The Bioinformatics revealed that the homology showed disparation form different rabies vaccine virus. the full-length gene sequence of rabies vaccine virus aG strain provided a support for perfecting the standard for quality control of virus strains for production of rabies vaccine for human use in China.
Amino Acid Sequence ; Antigens, Viral ; genetics ; immunology ; Base Sequence ; China ; Genome, Viral ; genetics ; Genotype ; Humans ; Molecular Sequence Data ; Phylogeny ; Rabies ; immunology ; prevention & control ; virology ; Rabies Vaccines ; immunology ; Rabies virus ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Species Specificity

BACKGROUNDTo determine if the attenuated Japanese encephalitis (JE) virus SA14-14-2 vaccine strain interacts efficiently with Culex tritaeniorhynchus and Culex pipiens quinquefasciatus, and further to acquire a new knowledge of its characteristics and safety for human beings.

METHODSLaboratory colonies of the two species of mosquitoes were set up and were inoculated intrathoracically with the attenuated vaccine virus and wild JE virus (Nak), both of which were used with different dilution from 10(-1) to 10(-9). Subsequently, the virus titers in the mosquitoes were detected by the plaque assay.

RESULTSInoculated with the vaccine strain, two species of mosquitoes were infected with the titers ranged from 10(0)-10(-3), and the maximum titers in Culex tritaeniorhynchus and Culex pipiens quinquefasciatus were 4.48 logPFU/ml and 5.63 logPFU/ml, respectively. Inoculated with wild JE virus, Culex pipiens quinquefasciatus was infected with titers ranged from 10(0)-10(-5), and the maximum titer in the mosquitoes was 6.59; Culex tritaeniorhynchus was infected with titers ranged from 10(0)-10(-4) and the maximum titer was 5.74 logPFU/ml.

CONCLUSIONBy intrathoracic infection, the attenuated JE virus SA14-14-2 vaccine strain can replicate in both species of mosquitoes.

Animals ; Culex ; classification ; virology ; Encephalitis Virus, Japanese ; genetics ; growth & development ; immunology ; Encephalitis, Japanese ; virology ; Humans ; Insect Vectors ; virology ; Japanese Encephalitis Vaccines ; immunology ; Species Specificity ; Vaccines, Attenuated ; immunology ; Viral Plaque Assay

OBJECTIVETo construct infectious Japanese encephalitis virus (JEV) based on the in vitro-ligated cDNA template of the vaccine strain SA14-14-2, and identify the virus.

METHODSFull-length genomic cDNA of JEV SA14-14-2 strain was ligated and then RNA was transcribed in vitro, the infective virus was obtained by transfecting the RNA into Vero cells and identified.

RESULTSThe infective clone of JEV was constructed, the virulence was weaker than the wild virus.

CONCLUSIONIt was possible to construct infectious clone from the production strain of live attenuated Japanese B encephalitis vaccine.

Animals ; Animals, Newborn ; Base Sequence ; Cells, Cultured ; Cercopithecus aethiops ; Cloning, Molecular ; DNA, Complementary ; genetics ; Encephalitis Virus, Japanese ; genetics ; immunology ; pathogenicity ; Encephalitis, Japanese ; pathology ; virology ; Genome, Viral ; Japanese Encephalitis Vaccines ; immunology ; Mice ; RNA, Viral ; genetics ; Vaccines, Attenuated ; immunology ; Vero Cells ; Virulence