Table of Content

30 August 2010, Volume 28 Issue 4

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论著

Two New Species of Freshwater Crabs （Decapoda ∶ Potamidae） Serving as Intermediate Hosts of Paragonimus in Fujian，China

CHENGYou-zhu;LINGuo-hua;LIYou-song

2010, 28(4):  1-245. 

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【Abstract】 Objective To investigate the natural resources of the freshwater crab which can serve as the intermediate hosts of Paragonimus in Pinghe and Youxi of Fujian Province. Methods Freshwater crabs were collected. The morphological characteristics of the crabs and the habitats were observed. The crabs were dissected and examined for the presence of Paragonimus metacercariae. Results Two new species of crabs were described, named as Sinopotamon zhangzhouense sp. nov. and Bottapotamon youxiense sp. nov.. S. zhangzhouense sp. nov., holotype:♂, carapace length 35.9 mm, breadth 42.8 mm, thickness 18.6 mm, collected from Pinghe County in southwest of Fujian. （24°14.206′N, 117°12.594′E）. Distal segment of the first pleopod of male tended flattish, and showed palm nest-shaped concave, which divided into two point leafs, and longitudinal crack clearly identified in back. The end half of distal segment reversed to ventral outwardly. This species usually lived in the sluggish stream. The infection rate of Paragonimus westermani and P. cenocopiosus in S. zhangzhouense sp. nov. was 44.9%（35/78）. B. youxiense sp. nov., holotype:♂, carapace length 13.35 mm, breadth 16.63 mm, thickness 7.20 mm, collected from Youxi County in central Fujian （26°10.558′N, 118°22.012′E）. The first pleopod of male was in slightly flat shape, ample and developed, as bow-like uplift. This species usually lived in the relatively flat terrain of stream. The infection rate of P. skrjabini metacercariae in B. youxiense sp. nov. was 92.1% （58/63）. Conclusion Two new species of freshwater crabs （S. zhangzhouense sp. nov. and B.youxiense sp. nov.） serving as the intermediate hosts of Paragonimus have been described.

Evaluation on the Immuno-protective Efficacy of the Recombinant Antigen SjPGAM-SjEnol against Schistosoma japonicum in Mice

GUOFan-ji;WANGYan;LIYe;PENGJin-biao;HONGYang;QIUChun-hui;CHENShi;FUZhi-qiang;SHIYao-jun;LINJiao-jiao*

2010, 28(4):  2-251. 

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Objective To construct and express the recombinant plasmid pET32a-SjPGAM-SjEnol and evaluate its immuno-protective efficacy against the infection of Schistosoma japonicum in mice. Methods The peptides of SjPGAM and SjEnol containing the multivalent epitopes with higher binding capacity of human MHCⅡand mouse H2-dⅡ but low homology with the host were analyzed and screened through bioinfomatics. The corresponding nucleotide sequence of selected epitopes was spliced and the recombinant plasmid pET32a-SjPGAM-SjEnol was constructed and expressed in Escherichia coli BL21 cells. The antigenicity of the recombinant protein was detected by Western blotting and the protective effect induced with the recombinant was evaluated in mice. 55 BALB/c mice were randomly divided into 5 groups each with 11. Mice from groups A, B and C were injected with a mixture of recombinant protein （27 μg） pET32a-SjPGAM-SjEnol （A）, pET28a-SjPGAM （B） and pET28a-SjEnol （C） respectively together with 206 adjuvant, mice from groups D and E received adjuvant or PBS only, all injected for three times at two-week intervals. Mice were then challenged with 40±2 cercariae of S. japonicum at two weeks after the last vaccination, and sacrificed for perfusion by 6 weeks post infection. Adult worms were collected, the number of eggs in a gram of liver tissue was counted, and the rates of worm reduction and egg reduction were calculated. Serum samples were collected before vaccination, every one week after each inoculation and before sacrifice, and specific IgG was detected by ELISA. Results The sequences encoding the 96-147 aa of SjPGAM and 233-312 aa of SjEonl were chosen for constructing the recombinant plasmid, a cDNA fragment with the length of 447 bp was amplified by PCR. The recombinant plasmid was expressed in E. coli with a molecular weight of M_r 33 000. Western blotting revealed that the fusion protein was recognized by the rabbit serum specific to SjSWAP, and showed an adequate antigenicity. Vaccination experiment showed that when compared with those of the blank control, the worm reduction rate in group A was 39.7%, significantly higher than that of groups B （18.5%） and C （14.7%） （P<0.05）. The liver egg reduction rate in group A was 64.9%, also higher than that of groups B （47.5%, P<0.05） and C （30.5%, P<0.01）. ELISA showed that the serum specific IgG in group A （2.372±0.268） was much higher than that of groups D （0.490±0.138） （P<0.01） and E （0.220±0.088） （P<0.01）. Conclusion The recombinant plasmid pET32a-SjPGAM-SjEnol has been constructed, and recombinant protein pET32a-SjPGAM-SjEnol induces higher immuno-protection against S. japonicum than that of SjPGAM and SjEonl.

Study on Molecular Phylogeny of Schistosoma bovis Based on Mitochondrial DNA Sequence and Gene Order

XIAOJing-ying;CAILian-shun;NagatakiMitsuru;TokuhiroShinji;JarillaBlancaR.;ShimadaMasaaki;DavidBlair;AgatsumaTakeshi*

2010, 28(4):  3-256. 

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Objective To determine the nucleotide sequence of the partial mitochondrial （mt） genome and the order of the mitochondrial protein-coding genes for Schistosoma bovis for analysis of possible phylogenetic position of this species in the genus Schistosoma. Methods　 The genomic DNA of adult worms were extracted by the GNT-K method. The target regions were amplified by PCR using a degenerated primer and specific primer. The PCR products were purified before ligating into the pGEM1 T-vector system. Recombinant plasmids were amplified in Escherichia coli， extracted and purified using routine methods. The nucleotide sequences were determined with an ABI PRISM 3100-Avant DNA sequencer using a BigDye Terminators v3.1 Cycle Sequencing Kit （Applied Bio-systems， CA， USA） with two T-vector specific primers （T7 and SP6）. Positive colonies were sequenced with two internal specific primers to obtain the full sequence of each fragment on both strands by means of primer walking. Sequences of related schistosomes were retrieved from GenBank and aligned with our data. Gene trees were constructed using neighbor joining methods. 　 Results　 The nucleotide sequence was determined and the gene order of this region in S. bovis was found as follows: NADHdehydrogenase4（nad4）-trnQ （Gln）-trnK（Lys） -NADH dehydrogenase 3（nad3）-trnD （Asp）-NADH dehydrogenase 1（nad1）. The gene order covering such region of S. bovis was similar to that of the African Schistosoma species，but strikingly different from the Asian species. Phylogenetic trees inferred from the alignment including partial nad4，nad3，partial nad1 and partial nad4+nad3+nad1 sequence for other 8 Schistosoma spp., respectivly， revealed that S. bovis is placed proximally to S. haematobium in the African sub-group， which is identical with those placed by gene order in the African clade. Conclusion The mtDNA analysis based on mitochondrial DNA sequence and the gene order strongly support the hypothesis that S. bovis belongs to the African schistosome clade rather than the Asian Schistosoma species.

Inhibition of Pyruvate Kinase mRNA Expression in Giardia lamblia by Specific Hammerhead Ribozyme

FENGXian-min;CAOLi-jing;WANGFeng-yun;ZHANGXi-chen;LUSi-qi*

2010, 28(4):  4-260. 

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Objective To inhibit the expression of pyruvate kinase （PK） mRNA in Giardia lamblia by specific hammerhead ribozyme. Methods The constructed hammerhead-GCV vector （pGCV-PKH） which aims to PK mRNA was electroporated into G. lamblia trophozoites （group A）. Electroporated trophozoites （group B） and normal trophozoites （group C） served as control. Trophozoites in each group were collected at 24, 48, 72 and 96 h post-electroporation, respectively. The concentrations of trophozoites were calculated and the growth curves were constructed. At 24，48，72 and 96 h post-electroporation, mRNA of each group was detected by RT-PCR and realtime PCR, respectively. The PK activity was tested by ultraviolet spectrophotometry. Results The growth curve showed that the growth of trophozoites was considerably depressed after 96 h post-electroporation. RT-PCR result displayed that the specific ribozyme mRNA was detected in group A from 24 h to 96 h post-electroporation. At 24 and 48 h after transfection, the PK mRNA level of group A decreased to 5% （5±0.17） and 8% （8±0.19） of the level in group C, respectively; and the PK activity of group A decreased to 32%（32±0.64） and 38% （38±0.65） of the level in group C. Conclusion PK mRNA expression in G. lamblia has been inhibited by specific hammerhead ribozyme.

Expression and Characterization of the FABP and Eg95Fusion Gene from Echinococcus granulosus

WANGLing;HUHui-xia;ZENGSu-xiang;WUWei-hua;GANWen-jia;HULi-ping;HUXu-chu*

2010, 28(4):  5-265. 

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Objective To construct and express a fusion gene of fatty acid binding protein （FABP） with Eg95 which are protective antigen genes of Echinococcus granulosus, and investigate the immunological characteristics of the recombinant protein. Method Using cDNA fragments encoding FABP and Eg95 genes from E. granulosus Qinghai sheep strain as templates, a fusion gene FABP·Eg95 was amplified by asymmetric polymerase chain reaction through a linking sequence encoding four glycine residues. The PCR products of fusion gene were subcloned into the pET28a （+） vector. The recombinant plasmid was transformed into Escherichia coli BL21（DE3） cells and induced with IPTG. The recombinant protein was detected by SDS-PAGE and purified by Ni-IDA affinity chromatography, and its immunogenicity was analyzed by Western blotting. Results The fusion gene length was about 795 bp. Double enzyme digestion analysis and DNA sequencing confirmed that the fusioin gene was cloned into pET-28a （+）. The recombinant protein （M_r 31 000） was expressed in inclusion body in E. coli expression system, and was purified by Ni-IDA affinity chromatography. Western blotting analysis testified that the recombinant protein could be recognized by sera of cystic echinococcosis patients, but not by sera of healthy persons and schistosomiasis patients. Conclusion The FABP·Eg95 fusion gene has been constructed, and the purified recombinant protein has been confirmed with immunogenicity.

Changes of Spleen Cytokines in Mice Immunized with Recombinant Bb-Eg95-EgA31 Vaccine against Echinococcus granulosus

ZHOUBi-ying;CHENYa-tang;LIWen-gui*;YANGMei

2010, 28(4):  6-272. 

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Objective To investigate the level of cytokines in spleen from mice immunized with recombinant Bifidobacteria bifidum（Bb）-Eg95-EgA31 vaccine of Echinococcus granulosus（Eg） and challenged with Eg protoscoleces. Methods 56 female BALB/c mice were randomly divided into 7 groups: Recombinant Bb-Eg95-EgA31 vaccine subcutaneous injection （group A）， intramuscular injection （B）， intranasal immunization （C）， oral administration （D）， blank vector control （E）， Bb control （F） and MRS control （G）. Mice in all groups were challenged with 50 Eg protoscoleces on the 8th week after vaccination and sacrificed on the 25th week after infection. Spleens were collected to prepare splenocytes, which were cultured under activation of crude Eg antigen （EgAg）， with concanavalin A （ConA） or lipopolysaccharide （LPS） stimulation and non-stimulation as control. The splenocyte supernatants were collected to determine the levels of interferon-γ（IFN-γ）， interleukin-12 （IL-12）， tumour necrosis factor-α （TNF-α） and IL-10 by ELISA. Results The level of IFN-γ in groups A ［（137.5±23.2） pg/ml］， B ［（162.5±23.2） pg/ml］， C ［（250.0±53.5） pg/ml］ and D ［（215.0±37.4） pg/ml］ was higher than that of group G ［（50.0±10.7） pg/ml］ （P＜0.01）. The level of IL-12 in groups A ［（27.5±4.6） pg/ml］， B ［（32.5±4.6） pg/ml］， C ［（45.0±5.4） pg/ml］ and D ［（35.0±5.4） pg/ml］ was higher than that of group G ［（15.0±5.4） pg/ml］ （P＜0.01）. The level of TNF-α in groups A ［（275.0±46.3） pg/ml］， B ［（325.0±46.3） pg/ml］， C ［（450.0±53.5） pg/ml］ and D ［（350.0±53.5） pg/ml］ was higher than that of group G ［（150.0±53.5） pg/ml］ （P＜0.01）. The level of IL-10 in groups A ［（37.5±4.6） pg/ml］， B ［（35.0±5.4） pg/ml］， C ［（25.0±5.4） pg/ml］ and D ［（32.5±4.6） pg/ml］ was lower than that of group G ［（45.0±5.4） pg/ml］ （P＜0.05 or P＜0.01）. Conclusion The recombinant Bb-Eg95-EgA31 vaccine can induce a protective Th1 type immune response in mice.

实验研究

Influence of Iron Ion on the Growth of Trichomonas vaginalis In Vitro

YUANYu-qing;XUEChang-gui*

2010, 28(4):  7-276. 

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Objective To study the influence of iron ion on the growth of Trichomonas vaginalis in vitro. Methods Quantitative pure incubation of T. vaginalis trophozoites with an initial density of 1×105/ml was carried out at 37 ℃ in TYM （trypticase-yeast extract-maltose） medium （pH 6.0） with additional 100, 200, 300, and 400 μmol/L iron ion, respectively, and in TYM medium without additional iron ion as control. The numbers of live and dead trichomonads were counted under an optical microscope after trypan blue staining, and the growth curve and survival rate of T. vaginalis were drawn to calculate the generation time of T. vaginalis. The minimal lethal concentration （MLC） of metronidazole was tested by serial dilution method when the trophozoites of T. vaginalis were incubated in TYM medium with 200 μmol/L iron ion and in control group without additional iron ion. Results The maximum densities of 2.9×106, 3.2×106, 3.1×106, and 2.8×106/ml were obtained when the trophozoites were incubated for 40 h in TYM medium with 100, 200, 300, and 400 μmol/L iron ion, respectively, while the maximum density was achieved at 50 h in the control, which was 2.5×106/ml. The generation time was （6.8±0.7） h in 400 μmol/L iron ion group, longer than those in the groups with 100-300 μmol/L iron ion, which were （4.8±0.3）, （4.8±0.2）, and （5.0±0.4） h, respectively （P﹤0.05）. While the generation times in 100-400 μmol/L groups were all shorter than that of the control ［（10.2±3.1） h］ （P﹤0.05）. MLC of metronidazole for T. vaginalis in TYM medium with 200 μmol/L iron ion was （23.44±11.56） μg/ml, significantly lower than that of the control ［（31.25±15.44） μg/ml］ （P﹤0.05）. Conclusion In TYM medium with 100-400 μmol/L iron ion, T. vaginalis trophozoites reproduce faster, and the MLC of metronidazole to the parasites is lower.

Effect of Six Anthelmintics in Oral Treatment of Mice Infectedwith Armillifer agkistrodontis Nymphs

XULi-li;XUEJian;ZHANGYong-nian;QIANGHui-qin;XIAOShu-hua*

2010, 28(4):  8-279. 

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Objective To observe the effect of praziquantel， mebendazole， tribendimidine， ivermectin， artemether and dihydroartemisinin against Amillifer agkistrodontis nymphs harbored in mice. Method Thirty-five mice infected each with 40 eggs of Amillifer agkistrodontis for 25-37 weeks were divided into 10 groups （2-8 mice per group）. Among them, nine groups were treated orally with praziquantel 500 mg/（kg·d）×5 d or 500 mg/（kg·d）×14 d， mebendazole 300 mg/（kg·d）×5 d， tribendimidine 300 mg/（kg·d）×5 d， ivermectin 8-10 mg/（kg·d）×3 d or 15 mg/（kg·d）×14 d， artemether 400 mg/（kg·d）×5 d and dihydroartemisinin 200 mg/（kg·d）×5 d， respepctively. The remaining untreated group served as control. All mice were sacrificed 1-3 weeks post-treatment for collection and separation of Amillifer agkistrodontis nymph cysts from abdominal wall， lipid tissue and viscera including liver， spleen and outer wall of intestine. The nymphs were released after tearing up the cyst with forceps, placed in the normal saline to observe their motor activity and counted. Results The difference of mean nymph burden between the drug treated groups and control group was not statistically significant （P>0.05） with mean nymph reductions of 8.3%-35.0%. Meanwhile， the appearance of the cyst， the size， colour， morphology and motor activity of nymphs were also similar to that of the control. Conclusion Praziquantel，mebendazole，tribendimidine，ivermectin，artemether and dihydroartemisinin exhibit no effect against Amillifer agkistrodontis harbored in mice under the conditions in the experiment.

现场研究

Investigation on Transmitting Vectors of Visceral Leishmaniasis in Jiashi County, Xinjiang

GUDeng-an*;ZUOXin-ping;YisilayinOSMAN;LANQin-xian;JINChang-fa;ZHOUXiao-jun;WEIFu-rong;ZHANGYi

2010, 28(4):  9-283. 

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Objective To investigate sandfly vectors transmitting visceral leishmaniasis, including species and seasonal distribution in Jiashi county of Xinjiang Uygur Autonomous Region. Methods Sandflies were collected in the field, counted and identified. The specimens were dissected to analyze the gonotrophic cycle and to find infection of promastigotes. The resting places were observed by using oil-paper and sandfly-capturing trap. Results 4 540 sandflies were collected with 99.9% of Phlebotomus wui and only 0.1% Sergentomyia minutus sinkiangensis. On the seasonal distribution, the first peak appeared by the end of May and the first ten-day of June, and the second peak was in the middle of August. Observation showed that the activity of sandflies occurred mainly from 22：00 to 4：00, reaching to the maximum in the midnight. Analysis on the gonotrophic cycle revealed that Ph. wui was an exophilic species and appeared nocturnally for feeding with preference to human blood. Natural infection with promastigotes was found in 4 sandflies, more in the field than the residential area. Resting places included the aperture on the wall of livestock sheds and in the caves. Conclusion Ph. wui is the predominant species in Jiashi, with higher infection rate of natural promastigotes in the field and with two life generations annually.

Evaluation of Recombinant 29 000 Extra Membranous Protein forthe Immunodiagnosis of Schistosomiasis japonica

WANGPing;RENCui-ping;WANGTian-ping;SHENJi-jia*

2010, 28(4):  10-286. 

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Objective To evaluate the recombinant membranous protein 29 000 （rSj29） of Schistosoma japonicum in the immunodiagnosis of schistosomiasis by indirect ELISA. Methods 394 serum samples collected from two low-endemic villages in Anhui Province were tested by rSj29-ELISA and AWA-ELISA using adult worm antigen (AWA) of S. japonicum. Meanwhile all subjects were each tested by 9 examinations with 3 fecal samples by modified Kato-Katz method, and by indirect hemagglutination assay （IHA）. Results It was found that the positive detection rates of stool examination, IHA, rSj29-ELISA and AWA-ELISA were 4.8% （19/394）, 62.2% （245/394）, 68.3% （269/394）, and 89.9%（354/394）, respectively. There was no statistical difference between rSj29-ELISA and IHA （χ²=3.2， P>0.05） with the coin-cidence of 80.7%（318/394）. The coincidence of IHA, rSj29-ELISA and AWA-ELISA with stool examination was 42.6% （168/394）, 36.0% （142/394）, and 15.0% （59/394）, respectively. Conclusion The indirect ELISA with rSj29 antigen shows similar effect as IHA for the diagnosis of schistosomiasis.

信息报道

Analysis on the Award-winning Research Achievements by the National Institute of Parasitic Diseases

ZHANGGuo-qing;GUANYa-yi;ZHANGMin-qi;TAOBi-ying;TANGLin-hua

2010, 28(4):  11-289. 

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Objective To evaluate the predominance， characteristic and shortage of the scientific activities at the National Institute of Parasitic Diseases （NIPD）， through analysis on its award-winning research achievements. Methods Information was collected on the award-winning research achievements by the Institute in the last 45 years. Time trend，disease category， subject， award type， award grade， award level and project property of the award-winning research achievements were analyzed by using SPSS11.0 software. Results Totally， 78 research achievements were awarded 128 times in the last 45 years. 43.6% awards were at province and ministry level； 23.1% awards were at nation level. Awards involved in schistosomiasis， malaria， and kala-azar accounted for 33.3%， 28.2% and 12.8%， respectively. As for subject，preventive medicine， pharmacy and biology accounted for 28.2%， 18.0% and 16.7%， respectively； 82.1% of the awards belonged to applied researches. Conclusion NIPD has a strong capacity in scientific research. To get more achievements in future， it is essential to closely integrate the experiment research to disease control.

专家论坛

A Proposal Concerning Renovation of Chinese Terms for Cestodes with Special References to Metacestodes and Spargana

WENTing-huan*

2010, 28(4):  12-296. 

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Some of the Chinese names involved in the cestode life cycle were not in coincidence with that of the Latin terms and their definitions， and may be imprecise in application. The term of metacestode has been misled into defining that is the larval stage in intermediate hosts or the infective stage to definite hosts; there was no specific Chinese term for sparganum which used to express it with same meaning and word as plerocercoid; the terms of oncosphere， procercoid and plerocercoid including cercomer were incorrectly translated; same meaning of hydatid and hydatid cyst were vaguely read in Chinese literature and protoscolex is concerning to a larva in the relevent literature. A renovated unified system of naming the various stages and phases of cestode development including adult in Chinese was proposed.

综述

Progress on Molecular Biology of Pneumocystis ——for Hundred Years since its Discovery

LIUQiu-ying;SIXiao-bei;ANChun-li*

2010, 28(4):  13-300. 

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2009 marks the 100th anniversary of the first description of Pneumocystis，an organism that was initially classified as a protozoan and later in 1988，was phylogenetically reclassified in the fungal kingdom. The organism was ignored for its first 50 years but that has subsequently been recognized as an important pathogen in immunocompromised patients，especially patients with acquired immune deficiency syndrome（AIDS）. This paper reviewed recent progress on the molecular biology，involving in the life cycle and metabolism of the organism.

Research Progress on the Relationship between Three Kinds of Liver Fluke Infections and Cholangiocarcinoma

LIUGuo-xing;WUXiu-ping;WANGZi-jian;BAIXue;LIUMing-yuan*

2010, 28(4):  14-305. 

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iver fluke disease caused by Clonorchis sinensis, Opisthorchis viverrini and O. felineus is a food-borne zoonotic parasitic disease and widely prevalent in Asia. The definitive hosts including human beings get the infection by ingestion of raw or undercooked fish or shrimp infested with metacercariae. Long-term or severe infection of the flukes can lead to dysfunction of the liver, such as cholelithiasis, cholecystitis, and so on. Researches indicated that there is an etiology relation between the fluke infection and cholangiocarcinoma. This paper reviews the relationship and possible mechanisms of the liver fluke-associated cholangiocarcinoma.

Application of PCR and Related Techniques in the Examination of Orientia tsutsugamushi

ZHANYin-zhu;GUOXian-guo*;ZUOXiao-hua

2010, 28(4):  15-312. 

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With the application of PCR technique in the medical field, PCR and its related techniques have made progress on detecting Orientia tsutsugamushi, which have played an important role in the early diagnosis of tsutsugamushi disease and pathogen research. This paper summarizes the application of PCR and related techniques in detecting O. tsutsugamushi．

震区防病

The Risk Evaluation and Response to the Spread of Hydatid Disease after Yushu Earthquake in Qinghai Province

WANGLi-ying;WUWei-ping*;LIShi-zhu;FUQing;WANGQiang;TIANTian;YANGShi-jie

2010, 28(4):  16-317. 

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Yushu of Qinghai Province is the mixed endemic area for cystic and alveolar hydatid diseases. Reviewing the previous data on the prevalence of the disease and the information from the communicable disease surveillance network during 2004 to April 15, 2010, possible impact of the Yushu earthquake was evaluated. It revealed that post-earthquake conditions might bring about risk factors for an increase of disease transmission, which included more deaths of livestock, increase of the number of stray dogs, poorer hygienic condition for local residents, etc. Countermeasures are proposed for preventing the spread of the disease in the disaster area.

研究简报

Genotyping of Plasmodium vivax in China-Myanmar Border by Circumsporozoite Protein

XUGui-Quan;ZHANGZai-xing*;LINa;DAIHong-jian

2010, 28(4):  17-267. 

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By using a series of type-specific primers for Plasmodium vivax circumsporozoite protein （CSP） gene and nested PCR, genotyping was conducted for the specimens of Plasmodium vivax isolated from China-Myanmar border. In 174 isolates of P. vivax, four genotypes, namely, tropical zone family strain, temperate zone family strain, genotype-mixed infection and PV-II type, were identified each accounting for 54.6 %, 35.6%, 6.9%, and 2.9%, respectively. The tropical zone family strain was dominant in the border area. There was no significant difference on the P.v CSP genotype constitution between Laza isolate of Myanmar and Tengchong isolate of Yunnan, China

Establishment of Loop-mediated Isothermal Amplification （LAMP） for Detecting Pneumocystis carinii

ZHANGHui;LIUXue-qing;HERong-zhi;JIATian-jun;ZHANGJin-shun*

2010, 28(4):  18-307. 

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Animal model of Pneumocystis carinii pneumonia （PCP） was established for acquiring lung tissue infected with P. carinii. After DNA from rat lungs was extracted， nuclear ribosome small subumit 18s rDNA of P. carinii was amplified by loop-mediated isothermal amplification method at 63 ℃ for 60 min. The product was digested by restriction enzyme ApalⅠ. The results showed that 18s ribosome DNA （rDNA） of P. carinii was cloned into vector pGEX6p2， and the positive clones screened. Therefore, the loop-mediated isothermal amplification has been established for detecting P. carinii.

Observation on the Ultrastructure of Spirometra mansoni Plerocercoid

TANGGui-wen;CHENGYan*;LIUXian-lin;CHANGAo-shuang

2010, 28(4):  19-314. 

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Plerocercoids of Spirometra mansoni were collected from muscles of the frogs. Specimens were treated following the routine procedure, embedded， sliced and stained. The ultrastructure of plerocercoid was observed with transmission electron microscopy. It was found that the wall of plerocercoid consisted of tegument and parenchyma. Thornshape microtriches distributed over the outer surface of the tegument. Matrix zone had a lot of granular discoidal bodies， vesicles， mitochondria and endoplasmic reticulum. Most of the mitochondria were near the basal membrane. Parenchyma zone consisted of muscular layer， tegument cells， parenchymal cells， excretory system, and so on. Many cytoplasmic pathways of tegumentary cells strech into muscular layer, suggesting that the tegument may be the absorptive site of nutrients.

In vitro Co-cultivation of Toxoplasma gondii Tachyzoites with Rat Brain Astrocytes

LIDong-na;LIANGYou-sheng;ZHOUYong-hua;ZHANGHuan-xiang;SHENGHai-ying;LUOWei;GONGWei;ZHUGEHong-xiang*

2010, 28(4):  20-320. 

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Purified astrocytes were cultured in plates. When astrocytes grew over 80% of the plate, tachyzoites of Toxoplasma gondii RH strain were added for co-culture. In the period of 0-72 h, change of the astrocytes and tachyzoites was observed after Giemsa staining. In 0-48 h, monodansylcadaverine （MDC） was used to study the action of autophagy in the process of tachyzoites invading astrocytes. At 1 h co-culture, tachyzoites had entered in astrocytes and the autophagosomes appeared. At 4 h, the autophagosomes increased pronouncedly. However, after 12 h, number of autophagosomes considerably decreased and damage of the cells occurred. 48 h later, autophagosomes disappeared and more astrocytes were destroyed. At 72 h most cells destroyed and tachyzoites were released. The result showed that autophagy is inhibited when the astrocytes were in vitro infected by tachyzoites.

病例报告

Two Cases of Thelaziasis

LiLiu-ping*

2010, 28(4):  21-276. 

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Two Imported Cases of Visceral Leishmaniasis in Kunming

DENGZhi-jie;LINJia

2010, 28(4):  22-封二. 

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Radiotherapy In a Case of Osteohydatidosis

BAOYong-xing;MAORui;XIEZeng-ru;WENHao*

2010, 28(4):  23-封三. 

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