[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.

[New animal hosts of rabies virus in mountain areas in Zhejiang province].

OBJECTIVE: To understand the prevalence of rabies among wild animals and the animal species in rabies epidemic areas of Zhejiang province. METHODS: One hundred and sixty samples were collected from the brain tissues of cats, stoats, Apodemus agrarius, Moschus chinensis, and Sus scrofa in Lishui and Chunan cities of Zhejiang province. Each sample was divided into four parts: cerebrum, mesencephali, cerebellum and gyms hippocampi which were used to determine the positive samples by detection of rabies virus specific antigens and nucleotides, using DFA and RT-PCR methods. RESULTS: Positive slides in the tests contained a glaring, apple green brilliance fluorescence using rabies virus specific monoclonal antibody against nucleoprotein. Using Nested-PCR method targeted at part of N gene, five positive samples were identified which consisting of four positive samples from stoats with positive ratio as 8.33% (4/48) and one positive sample from Apodemus agrarius with positive ratio as 1.75% (1/57). However, no positive result was found from cats, Moschus chinensis, and Sus scrofa samples. CONCLUSION: Rabies virus positive samples were identified from stoats and Apodemus agrarius in the mountain areas, with biological diversity in Lishui and Chunan cities of Zhejiang province, indicating that stoats and Apodemus agrarius might have played a role in human rabies and acted as host of rabies virus. In order to effectively prevent and control rabies virus under these complicated geographical and ecological environment, we must understand and evaluate the infection situation among animals in these regions.

[Complete genome sequencing and analyses of rabies viruses isolated from wild animals (Chinese Ferret-Badger) in Zhejiang province].

OBJECTIVE: Based on sequencing the full-length genomes of two Chinese Ferret-Badger, we analyzed the properties of rabies viruses genetic variation in molecular level to get information on prevalence and variation of rabies viruses in Zhejiang, and to enrich the genome database of rabies viruses street strains isolated from Chinese wildlife. METHODS: 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 N genes from Chinese Ferret-Badger, sika deer, vole, dog. Vaccine strains were then determined. RESULTS: The two full-length genomes were completely sequenced to find out that they had the same genetic structure with 11 923 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. CONCLUSION: The two 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 two full-length genomes, the similarity in amino acid level was dramatically higher than that in nucleotide level, so that the nucleotide mutations happened in these two genomes were most probably as synonymous mutations. Compared to the referenced rabies viruses, the lengths of the five protein coding regions did not show any changes or recombination, but only with a few-point mutations. It was evident that the five proteins appeared to be stable. The variation sites and types of the two ferret badgers genomes were similar to the referenced vaccine or street strains. The two strains were genotype 1 according to the multi-sequence and phylogenetic analyses, which possessing the distinct geographyphic characteristics of China. All the evidence suggested a cue that these two ferret badgers rabies viruses were likely to be street virus that already circulating in wildlife.

Proliferating progenitor cells: a required cellular element for induction of ischemic tolerance in the brain.

Permanent cerebral blood flow reduction results in brain injury (stroke), whereas transient ischemic stress results in preconditioning, which can ameliorate the extent of irreversible brain injury from subsequent ischemia-the phenomena of ischemic tolerance. Neurogenesis in the brain occurs after both ischemic injury and the brief ischemia resulting in preconditioning. As neurogenesis is regarded as having an intrinsic neuroprotective role in the brain, we investigated the possible role of these endogenous progenitor cells in the induction of ischemic tolerance. Methylazoxymethanol acetate (MAM) was injected in wild-type mice to attenuate precursor cell proliferation and ganciclovir was used to diminish newly generated cells in GFAP/HSV-TK mice. Both MAM and ganciclovir significantly attenuated ischemia-induced progenitor cell proliferation in the subventricular zone, dentate gyrus, penumbra, and corpus callosum as quantified by 5-bromo-2'-deoxyuridine- or Ki-67-positive cells. Attenuation of ischemia-induced progenitor cell proliferation in the brain blocked the induction of ischemic tolerance. Further the number of TUNEL (TdT-mediated dUTP nick end labeling)-positive cells was considerably increased in MAM-treated animals, whereas MAM did not cause cell death in sham-operated controls. The results of this study suggest a role for endogenous progenitors in the protective effect of ischemic tolerance.