Allograft inflammatory factor-1 alleviates liver disease of BALB/c mice infected with Schistosoma japonicum.

Allograft inflammatory factor-1 (AIF-1) plays an important role in various inflammatory conditions. Our previous study demonstrated that AIF-1 was over-expressed in the liver of BALB/c mice infected with Schistosoma japonicum and played significant role in the pathogenesis of schistosomiasis. The aim of this study was to focus on the effect of AIF-1 treatment on liver fibrosis and necrosis of BALB/c mice infected with S. japonicum. Seventy-two BALB/c mice were infected with cercariae of S. japonicum and then divided into three groups: AIF-1-treated group, saline-treated group, and control group. The vital signs, liver function, egg load, and hepatic pathological changes of the mice were assessed, and the levels of AIF-1 and TNF-α in the liver and spleen were measured at 5, 8, and 14 weeks postinfection. The treatment of AIF-1 on the mice infected with S. japonicum suppressed the expression of TNF-α and increased the effectiveness of AIF-1 in the liver and spleen at 14 weeks postinfection. Histopathological analysis and Masson trichrome staining for the liver tissues showed that the liver fibrosis and necrosis were alleviated previously compared with other infected mice at 14 weeks postinfection. The treatment of AIF-1 on the mice infected with S. japonicum can alleviate hepatic fibrosis and necrosis which indicate that AIF-1 use may prevent and cure the liver fibrosis.

Molecular phylogenetics of Triculine snails (Gastropoda: Pomatiopsidae) from southern China.

Triculine (Gastropoda: Rissooidea: Pomatiopsidae) snails are involved in the transmission of schistosomiasis and paragonimiasis; their distributions are mainly across southeastern Asia and southern China. In the present investigation, partial sequences of COI, 16S, and 28S were examined to infer the phylogenetic relationships among the species rich and poorly understood gastropod. Samples were collected from 12 geographic locations in six provinces of southern China. Several methods such as maximum parsimony, maximum likelihood and distance analysis were used in phylogenetic analyses among these taxa. The resultant phylogenetic trees showed a similar topology irrespective of the phylogenetic methods used. The taxa fell into two clades, with those from Fujian, Guangxi, and Zhejiang Provinces in one clade and those from Hunan, Sichuan and Hubei in the other. Among the taxa in Hubei Province, five formed a monophyletic clade, but Tricula sp. H-SHY fell into a sister clade of Tricula hortensis of Sichuan, whilst Tricula hongshanensis formed a single clade. Sister taxa Tricula pingi and Tricula hsiangi formed well-supported clade within almost all the trees. These results, while preliminary, represent the first attempt to reconstruct a phylogeny for Triculinae across China.

[Genetic variation of two mitochondrial DNA molecules from three isolates of Oncomelania hupensis].

OBJECTIVE: To study the genetic variation of two mitochondrial DNA molecules (CO1 and Cytb gene) of Oncomelania hupensis isolated from different areas. METHODS: Snails were collected from Jingxi of Guangxi, Yueyang of Hunan and Eryuan of Yunnan. Genomic DNA was extracted from the snails, Col and Cytb gene fragments were amplified by PCR, then purified and sequenced. Sequences of each isolates were edited by using Clustal W (1.82) software, and the nucleotide composition, transition and transversion were accounted by using MEGA (3.1) software. The genetic distances were computed with Kimura method and phylogenetic trees were constructed with UPGMA and MP method respectively. RESULTS: CO1 and Cytb gene fragments were about 700 bp and 600 bp (including 2 primers) respectively. A total of 106 mutation spots (15.9%) were tested in CO1 homological nucleotides, and 165 mutation spots (28.5%) were tested in Cytb homological nucleotides. The distance matrix between Guangxi isolate and Hunan isolate was 0.051 and 0.031 for CO1 gene and Cytb gene respectively; while that between Guangxi and Yunnan isolates was 0.158 and 0.405 respectively. Phylogenetic trees constructed by UPGMA and MP took on the similar topo-structure: isolates of Guangxi and Hunan clustered into one group, while the Yunnan isolate exhibited as another group. CONCLUSION: Oncomelania hupensis in Guangxi, Hunan and Yunnan are of relatively rich polymorphism in CO1 and Cytb genes in general.

[Studies on the protective effect of the mutant of Sj23 DNA vaccine against schistosomiasis].

OBJECTIVE: To investigate the protective immunity of the vaccine against schistosomiasis, a mutant of Mr 23 000 membrane protein DNA (Sj23DNA) without the homologous sequence of ME491. METHODS: The mutant of Sj23 DNA with no homologous sequence of ME491 on the cell membrane of human melanoma was obtained by overlap PCR. The mutant was transfected into human embryonic kidney cells of the line HEK293. Indirect fluorescent antibody test (IFAT) was used to detect the expressed protein. Expression of the mutant of Sj23DNA in muscular cells of mice was conducted through vaccinating the mouse with 100 microg purified plasmids by injecting them into the quadriceps muscle of thigh. Four weeks after the immunization, the quadriceps muscles were taken and cryostat sections were prepared for detecting the expression by IFAT. Forty BALB/c mice were randomly divided into four groups and injected with the mutant of pcDNA3-Sj23 plasmid DNA, pcDNA3-Sj23 plasmid DNA, pcDNA3 blank plasmid (100 microg per mouse) and sterile saline (30 microl per mouse) respectively. Four weeks after the immunization, mice were challenged with cercariae (40+/-2 cercariae per mouse) by abdominal skin penetration. Mice were then killed 6 weeks later, perfusion and squash methods were carried out to collect the adult worms and the number of eggs per gram of liver tissue was calculated. Worm and egg reduction rates were used to evaluate the protective immunity. RESULTS: Specific fluorescence was demonstrated in muscular cells of mice vaccinated with the mutant of pcDNA3-Sj23. The worm reduction rate and egg reduction rate were 40.3% and 42.8% respectively in the mutant of pcDNA3-Sj23 group, which were higher than those in the pcDNA3-Sj23 plasmid group (33.1% and 28.9% respectively). The difference between these two groups was significant (P<0.05). CONCLUSION: The modified Sj23DNA without the homologous sequence of ME491 induces higher protection against Schistosoma japonicum infection in mice than that of Sj23DNA.

[Study on the geographic information system databases regarding the control of schistosomiasis in Zhongxiang, Hubei province, China].

OBJECTIVE: Using geographic information system (GIS) and the remote sensing techniques (RS), we developed a schistosomiasis database and geographic distribution map in Zhongxiang city,Hubei province in order to display and analyze the endemic situation longitudinally after the water conservancy project is completed. METHODS: Epidemiological data of schistosomiasis and the correlated climate and hydrology data for the last 30 years were collected and the relevant GIS databases were established under Artificial Neural Networks(ANN) and network training of Landsat TM images. RESULTS: GIS database of schistosomiasis in Zhongxiang city, Hubei province and its vicinity areas were developed including 1 maps regarding the epidemic situation of schistosomiasis. The areas of snail distributing were 4.4 hm2, 8.2 hm2, 24 hm2, 130.4 hm2, 8.13 hm2 and 7.53 hm2, respectively. CONCLUSION: The maps created by GIS database and RS techniques supported the complicated query on space and property, providing a new way in keeping,updating and analyzing available data. The techniques used should be able to provide evidence for the control of schistosomiasis to this water conservancy project.

[Studies on the genetic variation of two mitochondrial DNA molecules of Schistosoma japonicum].

OBJECTIVE: To study the variation of Schistosoma japonicum through two mitochondrial DNA molecules. METHODS: Genomic DNA was isolated with kit, and the mitochondrial NADH dehydrogenase 1 (ND1) and cytochrome c oxidase I (COI) gene fragments were amplified by polymerase chain reaction (PCR) and sequenced. The gene trees were constructed and the acquired data were analyzed with the help of bioinformatics. RESULTS: The gene trees showed that the Taiwan isolate and the mainland isolates can be divided in two groups: a group from the hilly region (Yunnan and Sichuan), another group from the lake region (Hunan, Jiangxi and Anhui); isolates from Hubei are at a different position on the gene trees. CONCLUSION: There are variations among the geographic isolates of Schistosoma japonicum in China, nevertheless, they have close kinship.