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Objective: Analysis the epidemiological characteristics on rabies cases occurred in Guangxi from 2004 to 2008 and summarize the result of healthy-dog infection rabies virus investigation from 2006 to 2008. Exploring the high-occurrence and correlated factors on rabies in Guangxi., Methods: Data collected from the National Disease Surveillance System and the National Active Surveillance System for Rabies from 2004 to 2008 and Data of healthy-dog infection rabies virus investigation from 2006 to 2008 were analyzed., Results: The total rabies cases were 2463 in Guangxi from 2004-2008 and average incidence rate was 0.98 per 100 thousand per year. There were 95 counties had rabies case reported, anyway more than 10 cases occurred county number was declined while less than 5 cases rose year by year. The rabies case incidence area was expanded and the cases in middle and west area of Guangxi rose significantly. Rabies cases were reported whole year and no seasonal peak. Human rabies cases mainly were farmers, students and children. Yanger than 20 years old and elder 40 years old were the highest age groups in the population of the investigation, 83.79% cases were attacked by dogs. 78.5% cases classification category III. 83.17% cases had exposed on the upper and lower limbs, 10.56% exposed to the head, face or neck. But 67.88% cases did not receive any PEP and only 18.31% cases vaccinated and 3.63% category III exposure cases combined administration of RIG. The incubation median was 60 days. The rabies virus infection rate among randomly collection healthy-dog brain samples from 2006 to 2008 was 1.92%, 0.93% and 0.89%., Conclusion: Unsuccessful and inadequate PEP of patients were the main factors leading to the high-occurrence of human rabies in Guangxi. And there are a lot of infection rabies virus healthy-dogs alive in Guangxi also as a high-occurrence factor.

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.

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.

To construct a expression plasmid containing the full-length cDNA of rabies virus, four overlapped fragments covering full length cDNA of rabies virus street stain HN10 were cloned into pVAX1 sequentially in the genome except for the G-L noncoding region which was replaced with GFP gene. The plasmid containing the full-length viral cDNA was flanked by hammerhead ribozyme (HamRz) and hepatitis delta ribozyme (HdvRz) sequences and arranged under the control of the cytomegalovirus (CMV) promoter. The constructed plasmid could be directly used for the following procedure of producing the recombinant rabies virus street HN10.

Objective: To understand the source and distribution of rabies virus (RV) in Hunan province with viral surveillance in order to provide scientific measures for prevention and control on rabies., Methods: Brain samples from healthy-looking domestic dogs were collected in the agricultural markets at the districts of high, middle, and low incidence rates and detected by direct Immunofluorescence assay (DFA). Positive samples would be further detected by RT-PCR and the surveillance samples were detected by RT-PCR. The positive samples detected by RT-PCR were sequenced with N gene., Results: The infection rate of those healthy-looking domestic dogs with rabies virus was 2.78% in Hunan province in 2005. 23 positive samples' N gene were sequenced and their similarities were 88.8% - 100.0%. The results indicated that Hunan rabies virus N gene aberrance was mainly synonymous aberrance and did not carry obvious regional characteristics. The rabies virus were circulating among different districts in Hunan province, and the neighboring provinces such as Guizhou, Hubei, Guangxi, Jiangsu and Henan. There were no positive samples detected in salivary, blood and urine samples. There was one positive sample detected in two skin samples., Conclusion: There are dogs infected with rabies virus found in Hunan province and this study showed that rabies virus detected in Hunan had a close genetic relationship with those rabies identified in other provinces, suggesting that study on the immunity and management of dog related rabies should be strengthened.

Objective: To understand the epidemiological features of two rabies cases in Baoshan city year 2006 and 2007 and to analyze its source of infection., Methods: Questionnaires were used to do the epidemiological survey on each of the rabies cases. Brain tissue samples of rabies patients were collect to detect the rabies virus by direct immunofluorescence assay (DFA) and RT-PCR assay. Homology and phylogenetic tree were analyzed, based on the whole nucleotide and deduced amino acid sequence of P, M and N gene of rabies virus followed by molecular epidemiological analysis., Results: In July 2006, one human rabies case was identified in Longyang district, and another one in Tengchong county in Baoshan city in 2007. The degrees of exposure of these two patients was all at degree III. Two brain tissue samples among the dead patients (No. CYN0601H and CYN0701H) were confirmed positive by both DFA and RT-PCR assay. The homology analysis of P, M and N gene sequences among CYN0601H, CYN0701H and other rabies strains isolated from other provinces and other counties, showed that the samples in Baoshan city shared the highest homology with the strains in Thailand. Phylogenetic analysis indicated that the two samples were very close and all belonged to genetype 1 Lyssavirus, with the closest relationship between samples in Baoshan city and strains in Thailand., Conclusion: It was confirmed on the virus molecular level that the two patients in Baoshan city were both suffered from rabies. The prevalent strains in Baoshan city was probably imported from foreign country, suggesting that prevention and control measures on rabies virus in the boarder areas of Yunnan should be strengthened.

Objective: Evaluation of the direct rapid immumohistochemical test (DRIT) for laboratory surveillance of rabies., Methods: 72 brain specimens of domestic dogs or patients collected from Guizhou, Guangxi, Hunan, Anhui, Jiangsu and Yunnan provinces were detected by conventional methods including Direct Fluorescent-antibody Assay (DFA) and Reverse Transcription Polymerase Chain Reaction (RT-PCR), and by DRIT which was newly developed in the Rabies Section of the Centers for Disease Control and Prevention in the United States. The sensitivity and specificity of DRIT were evaluated by compare of the three results. By analysis of the index including cost of experiment, technique requirement and so on, the advancement and applicability of DRIT were discussed., Results: Compared with DFA and RT-PCR, DRIT will be more applicable for laboratories with limited funds and weak techniques because of its lower cost needed and simpler techniques required while its sensitivity and specificity are equal to the other two methods., Conclusion: DRIT is more valuable in rabies diagnosis and more applicable for extension and popularization in rabies laboratory surveillance in local CDC.

Objective: Characterization of RdRp gene (L) of rabies virus aG and CTN181 strain in China., Methods: Overlapped fragments were amplified and assembled. Then characterization and phylogenetic analyses as well as prediction of functional regions were performed using biologic softwares., Results: L gene of CTN181 and aG strains were composed with 6387nt and 6384nt respectively and there were two repeated ATG in the start of L gene in CTN181 strain. In addition, some mutations and new functional regions were discovered and presumed to be crucial to the role of these regions in replication of rabies virus. Analyses of L gene provide new insight into the functional regions, and phylogenetic analyses showed that human vaccine strain aG were independent to others recommended by WHO., Conclusions: The characterization of complete L genes will be helpful for research on new functional regions, antiviral drugs targeted at RdRp and phylogenetic analyses.

Objective: Characterization of rabies virus phosphoprotein through analyzation of genetic variations about rabies virus phosphoproteins in high-incidence regions in China., Methods: The nucleotide sequence of the P gene of Guangxi, Guizhou and Hunan provinces positive sample's were sequenced, and the P region's similarity and phylogenetic analyses were completed by using softer wares., Results: The similarity of P region's nucleotide sequence is 82.1%-100%, while, the similarity of amino acid sequence is 87.5%-100%. A little variation in phosphoprotein cannot influence its biological functions., Conclusion: All rabies viruses isolated from Guangxi, Guizhou and Hunan provinces belong to genotype 1 and share same phylogenesis and same genome characteristic; Virus distribution presents unique Characterization; Some virus isolates from Hunan province and Thailand may come from the same virus.

Objective: To investigate the situation of dog rabies and analyze it's relationship with human rabies., Methods: In Guizhou, Guangxi and Hunan provinces which suffered from rabies most heavily, one or two cities were selected respectively from regions with high-, middle-, low-incidence rate of human rabies as investigation spots where Dogs' brain specimens were collected and detected with both direct immunofluorescence assay (IFA) and RT-PCR., Results: A total of 2887 specimens were collected and 66 of these were positive for IFA and RT-PCR. Therefore, the rate of positive specimens was 2.3%. However, there was not a close relationship in the incidence rate of dog rabies and human rabies., Conclusion: Dog's infection situation of rabies contributed to the severe epidemic of human rabies.

Objective: To observe the therapeutic effect and influential factors of strong stimulation of acupoints around eyes by reducing method of twisting the needle for treatment of adolescent myopia, and to compare the different curative effects of both weak and strong stimulus intensities and wearing glasses in daytime., Methods: According to the principle of knowing and permission, the patients were divided into a strong stimulation group (60 cases, 120 eyes), a weak stimulation group (60 cases, 120 eyes) and a wearing glasses group (60 cases, 120 eyes). Both the strong stimulation group and the weak stimulation group were treated by reducing method of twisting the needle, with Cuanzhu (BL 2), Yuyao (EX-HN 4), Sizhukong (TE 23), Sibai (ST 2) and Taiyang (EX-HN 5) selected. Each acupoint was twisted 360 and 90 times per min for the strong stimulation group, and 90 degrees and 60 times per min for the weak stimulation group; the wearing glasses group were not treated with acupuncture, but with wearing glasses in daytime., Results: After treatment with the reducing method of twisting the needle, the vision improved (P < 0.01), with more obviously increased in the strong stimulation group than the weak stimulation group (P < 0.01), and there was no significant change of vision in the wearing glasses group. The age and the myopic degree of the patient before treatment were closely related with the change of vision after treatment (P < 0.01)., Conclusion: The reducing method of twisting the needle has a definite clinical therapeutic effect on adolescent myopia; strong stimulation has a better effect than the weak one; the age and the myopic degree of the patient possibly influence the therapeutic effect.

Objective: To study genetic damage of mice caused by the volatile organic compounds (VOC) of decoration materials., Methods: Fifty-five hotel guest rooms newly decorated within 6 months and 18 hotel guest rooms not decorated within 3 years were selected to determine the concentrations of 6 main VOC (benzene, methylbenzene, dimethylbenzene, ethyl acetate, butyl acetate, formaldehyde) in the air. Mice were exposed to VOC with the concentrations of 5, 10, 20, 40 times respectively as high as those present in the newly decorated rooms in an exposure cabinet for 15 days. DNA damage of peripheral lymphocytes of the mice was determined by single cell gel electrophoresis (SCGE) and bone marrow micronucleus test., Results: The concentrations of benzene, methylbenzene, dimethylbenzene, ethyl acetate, butyl acetate and formaldehyde in the rooms newly decorated within 6 months (6.50, 3.00, 6.70, 41.33, 1.70 and 0.14 mg/m(3) respectively) were significantly higher than those in rooms not decorated within 3 years (0.08, 0.94, 1.38, 0.25, 0.25, 0.01 mg/m(3), P < 0.01). DNA damage rates of peripheral lymphocytes in the concentrations of 10, 20, 40 times of exposure groups were significantly higher than those in the control groups (P < 0.05 or P < 0.01), and the frequencies of micronucleus in the mice exposed to 40 times of concentration was significantly higher than that in control group., Conclusion: High concentrations of the volatile organic chemical compounds may cause genetic damage in mice. SCGE test is more sensitive than micronucleus test.