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β?carbonic anhydrases(β?CAs)are ubiquitous metalloenzymes which active site contains a zinc ion(Zn2+),and they could catalyze the hydration of carbon dioxide to bicarbonate and protons efficiently and are involved in many biological pro?cesses,such as respiration,pH and CO2 homeostasis,biosynthetic reactions,virulence regulation and so on,and may play a critical role in the life activity of many organisms which contain these enzymes. β?CAs are widely distributed in fungi,bacteria, algae,plants and a small number of protozoan and metazoan except vertebrates. Therefore,as potential drug targets for design?ing and developing antibacterial and anti?parasitic drugs,β?CAs promise a broad application prospect. This paper focuses on the distribution,physiological function and the progress of researches on β?CAs in parasites and their vectors.
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ABSTRACT:To explore the feasibility of DNA barcoding in the identification of sandflies ,the mitochondrial cytochrome c oxidase subunit I (COI) genes of 9 dominant species in China from three genera in the Phlebotominae were studied .The in‐traspecies and interspecies genetic distances were calculated using the Kimura‐2‐parameter model ,and the phylogenetic trees were reconstructed by neighbor‐joining (NJ) method .Results showed that all of sandflies species were successfully distinguish‐ed by the phylogenetic trees .The average intraspecific genetic distance (0 .8% ) was much less than the average interspecific ge‐netic distance (11 .2% ) .The study based on COI sequences indicates that the sandflies from China could be well distinguished with the commonly used COI barcodes ,which are potentially well utilized in sandflies species identification .
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The paper selectively introduces the famous and richly historical parasite collections in the world.Over the years these institutions or museums have proven to be valuable resource for researchers and students around the world for locating information about specimen holdings,accessions,category of type specimens for each collection listed,services(including resource sharing),and background of the reservation institutes.The information presented here could provide useful references for researchers and teachers.
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In order to get more nutrition from outside the erythrocyte,new channels were induced by malaria par-asite.These channels play an important role in physiology of the parasitized cell.They are of interest both as potential targets in their own right and as potential drug targeting routes capable of mediating the entry of cytotoxic drugs into the app-ropriate compartment of the infected cell.It is hoped that this new anti-malarial strategy will help to create a sustainable anti-malaria-drug-development portfolio for the treatment of malaria.
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This paper reports the rectification results of the tribe aedini mosquitoes formerly recorded in China, using the classification system proposed by Reinert during the recent years. Among all the 171 species of Chinese aedini mosquitoes examined, 160 species could be included in the new classification system. The other 11 species were listed in traditional taxonomic status for further study. The proposed new classification system of the Chinese aedini mosquitoes contained 29 genera, i.e. Aedes, Armigeres, Ayurakitia, Bothaella*, Bruceharrisonius*, Christophersiomyia*, Collessius*, Danielsia*, Downsiomyia*, Edwardsaedes*, Finlaya*, Fredwardsius*, Gilesius*, Heizmannia, Himalaius*, Hopkinsius*, Hulecoeteomyia*, Jihlienius*, Kenknightia*, Luius*, Mucidus*, Neomelaniconion*, Ochlerotatus, Phagomyia*, Scutomyia*, Stegomyia*, Tanakaius*, Udaya, and Verrallina. Among them, 22 genera (*) were new records in China. Besides, the authors made a significant revision to the following 4 species recorded formerly in 《Fauna Sinica, Insecta Vol. 8, Diptera: Culicidae》: Ae. (Edw.) antuensis as the synonym of Ed. pingpaensis, while Ae. (Sin.) occidentayunnanus, Ae. (Och.) flavidorsalis, and Ae. (Fin.) subsimilis should be rectified as Hz. (Mat.) occidentayunnana, Oc. albineus, and Ud. subsimilis, respectively.
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Objective To examine the inhibitory effect of antisense nucleic acid on the in vitro translation of esterase mRNA from dipterex resistant Culex pipiens pallens . Methods 18 mer nucleic acid was synthesized and complementary to the translation initiation site of mRNA of dipterex resistant mosquitoes. The ODNs were annealed to the corresponding mRNA molecules and they were added to rabbit reticulocyte cell free system. The translation products were analyzed by SDS PAGE. After fixing, the gel was exposed to X ray film by autoradiography for analysis of protein synthesis. Results Six ?mol/L of ODNs elicited a 50% reduction in specific protein expression , and 20 ?mol/L of ODNs inhibited the expression of esterase by 80%. The SDS PAGE showed that the band of reduced amounts of 65 kDa protein for resistant mosquito was almost the same as that for sensitive sample. Conclusion Antisense oligonucleic acids to the esterase mRNA of dipterex resistant mosquito could effectively inhibit its in vitro translation.
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Objective To investigate the mosquito abundance and their relative species composition within and outside the Rare Birds National Nature Reserve of Yancheng,Jiangsu Province. Methods Sampling was carried out between May and Oct. 2004 at two weeks interval in two foci (the Reserve and nearby residential district) in Sheyang County. Mosquitoes were collected with the modified CDC light trap. Density was calculated,and species were identified. Environmental temperatures,rainfall and relative humidity were monitored during the study. Results A total of 40 912 mosquitoes were captured in the two foci. The sampled mosquitoes were identified as 4 species belonging to three genera (Anopheles sinensis,Culex pipiens pallens,Cx. tritaeniorhynchus,and Armigeres subalbatus). The most abundant mosquito species was An. sinensis and Cx pipiens pallens,which accounted for 97.7% of the whole number. 92% and 8% of the total amount of mosquitoes were collected from the nature reserve and residential district respectively. The most abundant species in the nature reserve and residential district was An. sinensis(60.6%) and Cx. pipiens pallens(76%),respectively. Within the nature reserve,there were two peaks occurred in adult abundance,in mid-and late July and mid-Sept. The abundance of mosquitoes in the area was positively correlated to the temperature (r=0.765,P=0.005). Conclusion The wetland is an ideal breeding place for An. sinensis and Cx pipiens pallens. The peaks of mosquito abundance are in mid-and late July and mid-Sept. It is of importance to carry out surveillance on mosquito vectors with pathogen-transmitting potential.
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The isolation and culture of pathogenic free-living amoebae are useful in the diagnosis and research. This review focuses on the methods of isolation and cultivation of pathogenic free-living amoebae, including sample treatment, culture conditions, passage culture, pathogen detection, and maintenance.
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This paper presents a new revised "Checklist of the Anopheline Mosquitoes in China" based on the development of the mosquito-taxonomic researches during the years of 1988-2007. The new checklist contained 61 species (subspecies) of anopheline mosquitoes all in China. Twelve species among the past records were omitted because of their invalid specific names which were allocated into following categories: ① A doubtful record in China, with no typical specimen up to date since last century, e.g. Anopheles campestris reported in Yunnan; ② Misidentification: An. atroparvus and An. indiensis; ③ Confirmed as synonyms by hybridizing experiments or molecular identification, including 9 species as follows: An. changfus, An. dazhaius, An. kiangsuensis, An. anthropophagus, An. kunmingensis, An. xiaokuanus, An. junlianensis, An. yutsushiroensis (part) and An. fluviatilis. Meanwhile, the following rectified 4 anopheline mosquito species should be added to the new checklist: An. belenrae, An. lesteri, An. pullus, and An. baimaii.
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Objective To confirm the diagnosis of a human case with atypical vivax-malaria from Yunnan Province by molecular technique. Methods DNA was extracted from blood films of unidentified sample, and of four known Plasmodium species (P. vivax, P. falciparum, P. knowlesi, and P. cynomolgi). A DNA-based diagnosis with the polymerase chain reaction (PCR) method targeting the small subunit ribosomal RNA (SSU rRNA) genes of genus- and species-specific (two human malaria species and P. knowlesi) was introduced. Results The PCR amplification with primer pair specific for P. knowlesi produced a single fragment of 150 bp. Sequence analysis showed that the amplified fragment was identical to the sequence of P. knowlesi. Conclusion The patient was naturally infected with P. knowlesi.
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Objective To identify the recombinant aldolase (ALD) of Plasmodium falciparum, and to develop monoclonal antibodies (McAbs) against the recombinant ALD. Methods ALD gene was amplified by PCR from genomic DNA of FCC1/HN strain, and expressed in E.coli DH5?. BALB/c mice were immunized with the recombinant ALD of P. falciparum via celiac injection for 3 times with 2 weeks interval. Three days after a booster injection, spleen cells of the immunized mice were used for producing McAbs. The immune serum was tested by IFAT and Western blotting. Results BALB/c mice immunized with purified aldolase protein developed strong immune response to the antigen, and the titer of specific antibody reached 1∶105 in all immune sera after the third immunization. Moreover, immune sera specifically recognized the cultured P. falciparum. Western blotting showed that the immune sera recognized specifically a Mr 41 000 band of crude malaria antigen. No cross-reaction with human red cells was detected. Seven positive hybridoma cell lines were obtained after 3 rows of selection. All the McAbs′subclasses belong to IgG1. IFAT showed that only 4 McAbs could recognize the cultured P.falciparum. Conclusion Plasmodial aldolase has been successfully expressed and purified, and the established hybridoma cell lines can secrete McAbs specific to the aldolase of P. falciparum.
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A blood film slide taken from a patient previously diagnosed as vivax malaria in Mojiang County, Yunnan Province, showing atypical forms. The ring forms had multinuclei, and the late trophozoites trended to form band. The schizonts and gametocytes were somewhat alike to Plasmodium vivax. PCR amplification confirmed that the patient was infected by P.knowlesi.
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Objective To develop and identify the monoclonal antibodies (McAbs) against Region II~+ motif in circumsporozoite protein of Plasmodium falciparum. Methods BALB/c mice were immunized with 12 peptides within Region Ⅱ~+ in circumsporozoite protein of P. falciparum. Spleen cells isolated from the immunized mice were fused with myeloma cell. After three times screening with ELISA, 3 positive hybridoma cell lines were obtained. Results ELISA test indicated that the McAbs reacted with recombinant circumsporozoite protein fragment containing tandemly repeat region and conserved Region II~+. IFA test showed that the McAbs recognized not only the sporozoites of P. falciparum, but also the sporozoifes of P. yoelii. Conclusion McAbs obtained can probe the Region II~+ motif in circumsporozoite protein of P. falciparum, which might also recognize that of other Plasmodium species.
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A study under laboratory control was conducted for 3 months to investigate the dynamic changes of local skin reaction and histopathology in guinea pigs exposed to the continuous and repeated bites of mosquitoes (Aedes albopictus)at 1, 6, and 24h after biting.Histopathologically, there were massive cumulations of basophils in the upper dermis of the infected skin, which coincided well with the degree of skin reaction. It would seem reasonable to suggest that hypersensitive guinea pigs show both immediate reaction and cutaneous basophil hypersensitivity.The occurrence of basophils in Skin reaction to mosquito biting, or to other blood-sucking dipterous biting has not previously been reported.
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Regular surveillance of malaria epidemics is fundamenta l f or malaria epidemiology.Malaria early warning system(MEWS)based on malaria sur veillance is a systematic framework for the prediction of malaria epidemics.The surveillance included environmental factors,transmission potential,risk factors ,morbidity and mortality,and weather condition.The MEWS will enable the authorit ies to make early and correct forecast of malaria and take proper preventive mea sures.
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Objective: To understand the morphologic foundation for differences in drug-resistance, virulence and immunity between chloroquine-sensitive (N) and chloroquine-resistant (RC) strains of Plasmodium berghei. Methods: The RC strain and the N strain were compared concerning the formation and morphologic features of digestive vacuoles and haemozoin with transmission electron microscope. Results: There was a single large digestive vacuole and multiple micro-single-mem-braned vacuole-like structures in the trophozoites of the N strain, and haemozoins centralized and fused during their schizo-gony were situated under the plasma membrane. Whereas there were few digestive vacuoles in the trophozoites of the RC strain, but with multiple micro-single-membraned vacuole-like structures instead. The RC strain formed obviously less hemo-zoins than that of the N strain and the hemozoins were not centralized and fused during the schizogony. Conclusion: The RC strain forms multiple single-membraned food vacuole-like structures in the trophozoites, and has different mechanism for detoxifying free heme with N strain and the features may be the foundation for the difference in drug-resistance, virulence, immunity between RC strain and N strain of Plasmodium berghei.