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    30 June 2012, Volume 30 Issue 3
    Cloning,Expression and Immunodiagnostic Analysis of Schistosoma japonicum Calcium-binding EF-hand Domain Containing Protein
    LU Yan1,XU Bin2,JU Chuan2,MO Xiao-jin2,CHEN Shen-bo2,FENG Zheng2,WANG Xiao-ning1,3 *,HU Wei2,3
    2012, 30(3):  1-165-169. 
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    Objective   To clone and expression Schistosoma japonicum calcium-binding EF-hand domain containing protein (SjEFCAB), purify the expressed protein, and evaluate its antigenicity and diagnostic value.   Methods   The positive clone screened from egg cDNA library was used as template to amplify the SjEFCAB gene by PCR. The target fragment was cloned into prokaryotic expression vector pGEX-4T-1. The positive recombinant plasmids were transformed into E.coli BL21 and induced by IPTG for expression of the protein. The recombinant protein was purified with GST-tag affinity chromatography. Western blotting was used to analyze the antigenicity. The purified protein was used as coating antigen for indirect ELISA to evaluate its diagnostic effect. Serum samples from patients with schistosomiasis japonica (78 cases),clonorchiasis sinensis (5 cases), cysticercosis (10 cases), paragonimiasis westermani (6 cases), trichinosis (9 cases) and healthy persons (50 cases) were examined.   Results   The recombinant plasmid pGEX-4T-1-SjEFCAB was constructed and the SjEFCAB recombinant protein (Mr 8 200) was expressed in E. coli. The soluble fusion protein was purified with affinity chromatography. Western blotting analysis showed that the recombinant protein was recognized by sera of infected rabbits and pooled sera of schistosomiasis japonica patients. The sensitivity and specificity of ELISA for diagnosis of schistosomiasis japonica were 82.1% (64/78) and 95.0% (76/80), respectively. The cross reaction with sera of clonorchiasis sinensis, cysticercosis, and trichinosis patients were 1/5, 1/10, and 1/9, respectively. There was no cross reaction with sera of paragonimiasis westermani patients.  Conclusion  The recombinant SjEFCAB antigen has potential diagnostic value for schistosomiasis japonica.
    Recombinant Expression and Immunologic Identification of Bradyzoite-specific Antigen BSR4 of Toxoplasma gondii
    CHEN Xing-zhi1 *, GUO Kai1, CHEN Yong1, LIU Li-li2, SHEN Ji-long2, JIAO Yu-meng1, FANG Qiang1, SUN Xin1
    2012, 30(3):  2-170-173. 
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    Objective  To express the recombinant BSR4 protein of Toxoplasma gondii Prugniaud (PRU) strain and study its immunologic characteristics.  Methods  The recombinant plasmid pET28a(+)-BSR4 was transformed into E. coli BL21, followed by expression of BSR4 induced by IPTG and its purification. The immunoreactivity of the recombinant protein BSR4 was analyzed by Western blotting with the sera from mice infected by PRU strain of T. gondii or normal mice as the first antibody. The 3H-TdR incorporation assay was performed to determine the proliferation of splenocytes in mice infected by PRU strain stimulated by BSR4, and the stimulation index (SI) was calculated. ELISA was used to evaluate the immunoreactivity of BSR4 protein, in which 20 sera from each of acute (anti-T. gondii IgGIgM+) and chronic (IgG+IgM) toxoplasmosis patients, and healthy people were tested.  Results  The recombinant BSR4 was induced and expressed. After denaturation, renaturation and purification, the soluble protein (Mr 45 000) was obtained and detected by anti-T. gondii serum with Western blotting. BSR4 in 1, 5, and 25 μg/ml induced the proliferation of splenocytes in mice infected by PRU strain with higher SI (1.13, 0.88, and 1.17) than that of control (0.46, 0.24, and 0.49, respectively, P<0.01). ELISA showed that the recombinant BSR4 specifically reacted with the sera of toxoplasmosis patients (IgG+IgM, 20/20), not with that of the acute cases (IgGIgM+, 0/20).  Conclusion  The recombinant BSR4 shows specific immunogenicity and immunoreactivity.
    Construction of Rapid Tagging Vector of Genes in Giardia lamblia
    JU Hong-mei,WANG Yi-hui,ZHANG Xia,WANG Yun-hua,LI Ya-jie*
    2012, 30(3):  3-174-178. 
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    Objective   To construct a recombinant vector for rapid gene tagging in Giardia lamblia. Methods  To obtain the recombinant vector pGL gdh-Neo with the Neo selection marker, the Neo gene was put under the control of gdh promoter by overlap PCR and inserted into pGEM-5zf. A DNA fragment containing multiple cloning sites (MCS) followed by triple hemagglutinin(3HA) coding sequences was synthesized and cloned into the pGL gdh-Neo to construct a recombinant vector pGL MCS-3HA-gdh-Neo. Giardia H2A gene was selected as a tagging gene to validate the effectivity of the recombinant vector pGL MCS-3HA-gdh-Neo. The histone H2A coding sequence was amplified by PCR, digested with EcoRⅠ and SpeⅠ, and inserted into MCS of pGL MCS-3HA-gdh-Neo. The resulting plasmid was then linearized and transfected into Giardia trophozoites. The H2A recombinant strain selected by G418 was analyzed by PCR,Western blotting and immunofluorescence. Results  A rapid tagging recombinant vector with multiple cloning sites and triple hemagglutinin (3HA) was constructed with a length of 4 260 bp. The H2A recombinant vector was transfected into Giardia trophozoites and integrate into the Giardia genome at the correct locus. The HA-tagged H2A protein was expressed with a molecular weight(Mr) of 16 900.  Conclusion  A rapid tagging recombinant vector of genes in Giardia lamblia, pGL MCS-3HA-gdh-Neo, has been constructed.
    Kinetic Distribution of Cyst-forming Chinese Isolate of  Toxoplasma gondii in Mice
    WANG Lin1,2, HUO Xing-xing1,2, CHEN Zhao-wu1, CHEN He1, YU Li1, SONG Xiao-rong1, Faustina Halm-Lai1,3, LUO Qing-li1, SHEN Ji-long1*
    2012, 30(3):  4-179-183. 
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    Objective  To determine the kinetics of infection and cyst formation in CD1 mice following oral infec-tion with cyst-forming Chinese isolate of Toxoplasma gondii TgCtwh1(genotype China 1, ToxoDB#9).  Methods  50 CD1 female mice were obtained from specific pathogen-free(SPF) mouse colony in the Vital River Laboratories(VRL), Beijing. Mice were randomly divided into 10 groups each with 5 mice. All mice but control were peroral gavage infected with 50 cysts (1×104 bradyzoites) of TgCtwh1 isolate of T. gondii isolated from Wuhan, China. Cysts were isolated from the entire brain of mice infected with TgCtwh1 by density gradient centrifugation over Fycoll-paque plus. Animals were orally inoculated with cysts on day zero, and peripheral blood, lymph nodes, heart, liver, and brain of infected mice were collected on days 2, 4, 7, 10, 14, 21, 35, 50, and 72 post infection. Five mice were sacrificed by cervical dislocation under anes-thesia at each time of collection, and the kinetic distribution was detected by fluorescence quantitative PCR and tissue in-oculation into fresh mice. The cyst formation at various intervals after infection was also observed, as was the number of the cysts in brains and the cyst-forming rate.  Results  The body weight of the mice lessened (3.650±0.252) g post oral  infection on day 7,  and the weight was progressively decreased between day 10 [(1.730±0.017) g] and day 14 [(-0.390±0.554) g] after infection (P<0.05).  In the brain tissue, cysts were first observed on day 21 post oral infection and the cyst-forming rate was 80%, and the average diameter of cysts was 20-40 μm. While on day 35 after infection, the cysts were formed in all infected mice(cyst-forming rate was 100%) and the average diameter was 50-60 μm. In chronicinfection, DNA copies of parasites were first detected in blood, heart, liver and lymph node at 3.51±0.152, 4.100±0.198、4.220±0.209 and 4.960±0.052 respectively on day 2, then in the brain on day 4 (3.800±0.154). During the early days of infection, the parasite burden in blood was progressively increased until days 7 (5.240±0.115) then gradually decreased and become undetectable on day 35. The burden of T. gondii in the heart and brain tissues increased significantly andreached their maximum on day 14 (5.640±0.214) and day 10 (5.790±0.060), respectively, and remained a stable level thereafter. Liver and lymph tissues reached their maximum on day 7(5.310±0.038)and day 10(6.200±0.152), then gradually decreased and become undetectable on day 50.  Conclusion  The parasitemia in mice infected with T. gondii cyst-forming isolate lasts for 21 d at least, and cysts are detected in brain on day 21.
    Efficacy of Albendazole Orally Administered at Different Dosages against Trichinella spiralis Encapsulated Larvae in Mice
    LI Run-hua1,PEI Yan-jiang2,LI Qi-chao2,HUO Jiang2,DING Yu2,YIN Guo-rong3 *
    2012, 30(3):  5-184-188. 
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    Objective   To evaluate the efficacy of albendazole (ABZ) orally administered at different dosages against Trichinella spirails encapsulated larvae in striated muscle in mice.  Methods  A total of 72 BALB/c mice were divided equally into 9 groups. Each mouse was infected orally with 50 T. spiralis encapsulated larvae. At the 29th day after infection, albendazole was each orally administered to the mice of the 8 groups with doses of 50, 100, 150, 200, 250, 300, 350, and 400 mg/(kg·d), respectively, once a day at fixed time for 6 d. The control group was untreated. Mice were sacrificed at the 7th day post administration. The encapsulated larvae in diaphragmatic muscle, jugomaxillary muscle and gastrocnemius muscle were examined with pellet method. The encapsulated larva that the capsule was complete and the larva inside curled naturally with clear structure was considered survived. The therapeutic effect was estimated on the average quantity of total, survival and dead encapsulated larvae per gram muscle, total worm reduction and survival worm reduction.  Results  The total worm burden and survival worms showed a decreasing trend and the numbers of dead worms increased in diaphragmatic muscle, jugomaxillary muscle and gastrocnemius muscle when the dosage of albendazole were 50-250 mg/(kg·d), but the number of larvae in the muscles remained similar when the dosage of albendazole was greater than 250 mg/kg·d. Compared with the control group, the total and survival worms in the muscles in 200 mg/(kg·d) and the greater dose groups decreased significantly (P<0.01). In 250 mg/(kg·d) group the total worm reduction in jugomaxillary muscle, diaphragmatic muscle and gastrocnemius muscle were 50.00%, 62.62% and 57.48%, and the survival worm reduction were 79.96%, 83.25% and 80.56%, respectively.  Conclusion  Orally administered to mice for 6 d, albendazole at 250 mg/(kg·d) is a suitable dose against encapsulated larva stage of T. spiralis in muscle.
    Effect of Feeding on Different Tissues on Larva Development of  Chrysomya megacephala (Diptera ∶ Calliphoridae)
    LI Xue-bo1, LV Zhou1, WAN Li-hua1 *, WANG Yao1, YAO Lan1, WANG Qing-shan2
    2012, 30(3):  6-191-195. 
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    Objective  To observe the effect of feeding on different pig tissues on the development of Chrysomya megacephala larvae.  Methods  About 200 larvae each were reared on four different substrates, i.e. pig′s brain, liver, muscle and a mixture of minced pork muscle and fat (7 ∶ 3) at a constant temperature of 25 ℃. Length and weight of larvae and pupae were measured at 12 h interval 16 h after eclosion. 10 larvae or pupae were collected each time. The time of development, mortality, and sex ratio of adults were recorded.  Results  Three replicated experiments showed that the larvae fed on liver grew slowly, time of reaching maximum length and weight was delayed for about 24-36 h, and the duration of larva development was longer than that of other groups (P<0.01). The mean maximal larval length in mixture group [(14.89±0.39) mm] was statistically shorter than that of brain group, muscle group and liver group, [(17.81±0.54), (16.94±0.43) and (17.14±0.27) mm, respectively] (P<0.01). The mean maximal larval weight in liver group [(73.5±6.8) mg] and mixture group [(63.0±5.4) mg] was statistically lighter than brain group [(91.2±7.5) mg] and muscle group [(86.3±7.3) mg] (P<0.01). The pupal length in mixture group was statistically shorter than that of other 3 groups (P<0.01). The pupal weight of mixture group and liver group was statistically lighter than that of brain group and muscle group (P<0.01). The larval and pupal mortality of mixture group [(9.8±3.1)% and (8.9±3.1)%] was statistically higher than that of brain group [(5.5±3.1)% and (4.6±1.5)%], muscle group [(4.7±2.2)% and (3.8±2.0)%] and liver group [(5.4±2.3)% and (4.8±1.7)%] (P<0.01). There was no significant difference in the sex ratio among the four groups (P>0.05).  Conclusion  The development duration of the larvae fed on liver is longer than other groups. The body length and weight of larvae and pupae fed  on mixture diet are less than other groups with higher mortality.
    Expression of Tumor Necrosis Factor-α and Caspase-3 Protein in Monocytes Adjacent to the Invaded Echinococcus multilocularis in Liver
    YANG Hong-qiang1,2, MA Shao-bo1, BIAN Zhi-yuan1, Li Jiang1, ZOU Hong3, ZHANG Shi-jie1,2, PENG Xin-yu1,2, CHEN Xiao-ping2*
    2012, 30(3):  7-201-205. 
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    Objective   To investigate the expression and its significance of tumor necrosis factor-α (TNF-α) and caspase-3 protein in monocytes adjacent to the invaded Echinococcus multilocularis in liver.  Methods  40 female Kunming mice were randomly divided into experimental group (n=20) and sham operation (control) group (n=20). Mice in experimental group were infected with 20% E. multilocularis suspension (0.1 ml per mouse) through abdominal opening injection in liver and the mice in control group were injected with equal physiological saline. The mice were sacrificed at 6 months post-infection for observing the growth and metastasis of E. multilocularis. Pathological changes were observed by HE staining. The expression of TNF-α and caspase-3 protein in hydatid cyst and metastasis tissue were detected by immunohistochemistry staining and the apoptosis of the monocytes was measured by TUNEL.  Results  After 6 months post-infection, E. multilocularis were spread over the liver of the mice in experimental group. Metastasis rate of lymph nodes was 45.0%(9/20). Infiltration of monocytes was observed around E. multilocularis in liver and lymph nodes with metastasis by HE staining. Immunohistochemistry showed that the positive expression rate of TNF-α and caspase-3 protein in monocytes was 100% and 100%, and 95% and 100% respectively around the cyst in experimental group, while the expression rate was only 5% and 0 respectively in the liver of the control mice (P<0.01). The monocytes showed significant apoptosis by TUNEL in experimental group with a positive expression rate of 100%, with a significant difference between experiment group and the control (P<0.01). Conclusion  In the process of alveolar Echinococcus infection, the high expression of TNF-α protein might be associated with the apoptosis of monocytes, which may inhibit the host immunological function.
    Preliminary Study on the Difference in Immune Protection between Echinococcus granulosus Recombinant Ferritin and Recombinant mMDH
    LIU Li-hua1,ZHAO Jia-qing2,JIANG Bo3,CHEN Chang-yi4, WANG Yuan1,
    2012, 30(3):  8-206-209. 
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    Objective  To evaluate the immunoprotective activity of the Egrecombinant ferritin and Egrecombinant mMDH proteins in mice.  Methods  Thirty ICR mice were divided into 3 groups and immunized by injection of adjuvantemulsified rEgferritin, rEgmMDH and PBS, respectively, in multiple spots at back, for 3 times with an interval of 2 wk. Two weeks after the last immunization, the mice of the 3 groups were infected intraperitoneally with 0.1 ml suspension containing about 1 500 Echinococcus granulosus (Eg)  protoscoleces. The mice were sacrificed 22 wk after infection and the Eg cysts were collected and measured. Spleens were taken for detecting CD4+ T cells and CD8+ T cells and ratio calculated.  Results   Eg cysts were found in 30% (3/10) of the mice in the rEgferritin group with 5 cysts altogether; cysts were received in all the mice in the rEgmMDH group with 118 cysts totally; and cysts were found in 7 of 9 mice in the PBS control with 35 cysts totally. The mice in the rEgferritin group showed an 84.7% protection but revealed no protection in the rEgmMDH group. The CD4+ T cells were significantly higher in the rEgferritin group than the control, but no statistical difference was found in CD8+ T cells and CD4+/CD8+ ratio between the 2 groups. There was no considerable change in the T cells and ratio in the rEgmMDH group compared to the control.  Conclusion  The Egrecombinant ferritin can inhibit the growth of Eg while the Egrecombinant mMDH seems promoting its growth in mice.
    Mycoplasma hominis Symbiosis and Trichomonas vaginalis Metronidazole Resistance
    WANG Pin-jia1,XIE Cheng-bin2 *
    2012, 30(3):  9-210-213. 
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    Objective   To investigate the relation of Mycoplasma hominis symbiosis and the resistence of Trichomonas vaginalis to metronidazole.  Methods  From November 2010 to July 2011, 160 isolates of T. vaginalis were collected from the genital tract secretion of gynecological out-patients at the Sichuan Provincial Hospital for Women and Children. The minimum lethal concentration (MLC) to metronidazole of these isolates was determined by an in vitro sensitivity assay with different concentration gradients of metronidazole (from 1 to 1 024 μg/ml), and M. hominis DNA in T. vaginalis was detected by polymerase chain reaction (PCR) technique with specific 16S rRNA primers. After clearance of M. hominis from the parasites by 32 μg/ml doxycycline, MLC was determined and compared with that before clearance.  Results  MLC of metronidazole in T. vaginalis ranged from 1 to 256 μg/ml, with 61.3% isolates (98/160) ranging from 1 to 8 μg/ml, 26.3% isolates (42/160) ranging from 16 to 32 μg/ml, and 12.5% isolates (20/160) ranging from 64 to 256 μg/ml. 61 isolates were PCR positive for M. hominis DNA in the 160 isolates of T. vaginalis. The M. hominis DNA positive rate was significantly higher in the T. vaginalis isolates with higher MLC than those isolates with lower MLC (P<0.01). However, when M. hominis was cleared by doxycycline from 8 isolates among the 61 ones, no change was observed in sensitivity of the isolates to metronidazole.  Conclusion  M. hominis symbiosis might be associated with the metronidazole-resistance of T. vaginalis.  However, it needs direct evidence.
    Cloning of AGO2 and Dcr-2 Gene Fragments and Analysis of Their Transcription Level in Different Developmental Stages of Aedes albopictus
    CAI Yan-li,ZHENG Xue-li*
    2012, 30(3):  10-214-217. 
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    Objective   To perform molecular cloning of the AGO2 and Dcr-2 gene fragments associated with RNA interference pathway of Aedes albopictus and characterize the transcription level of the two genes across all life stages of the mosquito.  Methods  The degenerate primers were designed based on the conserved regions of AGO2 and Dcr-2 amino acid sequences, and the AGO2 and Dcr-2 cDNA fragments were amplified from total RNA of a female mosquito by RT-PCR. The PCR products were cloned into pMD18-T vector and transformed into E. coli DH5α strain, and the positive clones were selected and sequenced, with the results for homology analysis by Blastx. The specific primers were designed according to the sequences of AGO2 and Dcr-2 from Ae. albopictus, which were used to investigate the transcription levels of these two genes from eggs, Ⅰ and Ⅱ instars larvae, Ⅲ and Ⅳ instars larvae, pupa, male and female mosquitoes by semi-quantitative RT-PCR.  Results  The AGO2 and Dcr-2 cDNA fragments obtained were 326 bp and 491 bp in length, with the Accession number of JQ764670 and JQ764671, respectively. The Blastx analysis showed that the AGO2 and Dcr-2 amino acid sequences shared 91% similarity to AGO2 of Ae. aegypti and 98% to Dcr-2 of Ae. albopictus. The transcription of AGO2 and Dcr-2 genes was detected in all life stages of Ae. albopictus, with the highest level of mRNA in female mosquitoes, which was 3.1 times and 15.5 times higher for AGO2 and Dcr-2 than in male mosquitoes, respectively, and significantly higher than other developmental stages (P<0.05).  Conclusion  The AGO2 and Dcr-2 cDNA sequences have been partially obtained and the hightest transcription level found in female Ae. albopictus, suggesting that AGO2 and Dcr-2 are the key genes of RNA interference in female mosquitoes.
    Impact of Pesticide Use on the Density of Anopheles anthropophagus and Malaria Incidence
    PAN Jia-yun1,ZHENG Xiang1 *,QIAN Hui-lin1,LIU Chao-fan1,GUO Chuan-kun2,JIANG Miao-gen3,CHEN Huan-lu4,GU Zheng-cheng1
    2012, 30(3):  11-218-223. 
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    Objective   To investigate the impact of the application of chemical pesticides on the distribution of Anopheles anthropophagus in rice fields and the malaria incidence.  Method  Twenty-four villages from 16 counties in the provinces of Zhejiang, Sichuan and Guangxi were chosen for the surveys in the period of 1983-1987. For the survey of An. anthropophagus, indoor human bait trapping until midnight and catching the mosquitoes in all the nets in early morning were carried out to get the density and population ratio of the mosquitoes. Historical data on Anopheles spp., malaria incidence, acreage of the single or double season cropping of rice and on the quantity of chemical pesticide used in rice fields were collected from the study areas. Results  In Hang-jia-hu region of Zhejiang Province, double season rice cropping was performed at that time, the quantity of pesticides used in 1973 was 45 kg/hm2, which was as high as 50 times than that in the 1950s. The density of An. anthropophagus decreased yearly, no An. anthropophagus could be found at 11 survey points in late 1980s. The malaria incidence dropped to less than 1 per 10 000. In Leshan and Yibin areas of Sichuan Province, the major cultivation was single cropping, pesticides were applied in paddy fields since 1960s, and the average quantity of pesticides used was 8.6 kg/hm2 during 1970s-1980s. No significant difference on the density of An. anthropophagus was revealed between 1980s (86.2%) and 1960s (82.2%)  (χ2=0.63, P>0.05). After mid-1980s, pesticide use gradually increased, and reached to 18.18 kg/hm2 in average in the years after 2000. The density of An. anthropophagus decreased, no An. anthropophagus was found in 2010 in the area surveyed and no malaria cases were reported as well. With double season cropping in Huanjiang County of Guangxi, the pesticide amount consumed was 1.79 kg/hm2, 25.13 kg/hm2 and 7.68 kg/hm2 in paddy fields in 1960s, 1970s, and 1980s, respectively. The proportion of An. anthropophagus in anophelines was 52%(1 747/3 392) in the beginning of the 1980s. After the year 2000, the average pesticide use increased to 20.38 kg/hm2 in paddy fields. It was difficult to find An. anthropophagus in human dwellings after 2008. The average annual malaria incidence dropped to 0.14 per 10 000.  Conclusion  Change of farming activities and especially use of chemical pesticides in high quantity at the rice fields undermine the breeding environments of An. anthropophagus, greatly reduce the mosquito population and therefore the malaria incidence.
    Pathogen Identification of 10 Suspected Cases  of Sparganosis mansoni
    ZENG Qing-ren1,HE Mei1,WANG Fang1,ZHANG Zu-ping1 *,SU Zhan-san2,ZHOU Jun2,LIU Bao-an3,LAN Zhi-hua2,HU Mian-juan1,CAI Li-ting1
    2012, 30(3):  12-224-227,232. 
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    Objective  To diagnose 10 cases of clinically suspected cases of sparganosis mansoni by pathogen identification.  Methods  In the period from August 2009 to August 2011, 10 biopsy specimens were obtained from 10 patients of four hospitals to identify the pathogen. Among the 10 cases, 4 cases showed abdominal subcutaneous mass, 3 showed eyelid swelling, 1 displayed brain lesions, 1 showed pulmonary mass, and 1 showed pleural effusion. There was one parasite each from three patients with eyelid swelling, and one patient with abdominal subcutaneous mass, which were observed by naked eye and microscope morphologically and histologically. Specimens from other six cases were examined by microscope after paraffin embedding, sectioning, and HE staining. For further identification, the parasite biopsy tissue specimens were detected by immunohistochemistry with Sparganum mansoni-immunized rabbit serum as the primary antibody.  Results  Three intact worms, from three patients with eyelid swelling, showed typical S. mansoni morphological characteristics. One residue parasite from the abdominal subcutaneous mass showed network structures and full of calcareous corpuscles in the body under microscope same as that of S. mansoni. The histological structure in three of the six sections showed typically the body wall with folds, which was dense, thick and deeply eosine stained, part of the tegument outside was covered by micro-hairs. In the worm body there was net-like loose structure and calcareous corpuscles without cavity. The structure of the other three worm sections was atypical. The six worm sections were positive by immunohistochemical detection.  Conclusion  The 10 clinically suspected cases are diagnosed as sparganosis mansoni.
    Research Advances of Interleukin-15 in Anti-parasitic Infection
    LI Jia-yuan1,2, LI Zhong-yuan1,2, WANG Yan1,2, ZHU Xing-quan1, XU Min-Jun1 *
    2012, 30(3):  13-196-200. 
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    Interleukin-15(IL-15) is a pro-inflammatory cytokine and also a chemotactic factor for a variety of immune cells, which participates in and regulates the inflammatory response and immune response to organisms. Up to now, the molecular structure and receptor of IL-15 have been widely researched, with important advances in its signal transduction way. However, its application in parasitology is still a relatively new topic. This article summarizes the latest research development of IL-15 in anti-parasitic infection, prospects its mechanism and application.
    Advances in Epigenetic Researches of Toxoplasma gondii
    YANG Pei-liang 1,CHEN Xiao-guang 2 *
    2012, 30(3):  14-228-232. 
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    Toxoplasma gondii undergoes a complex life cycle that involves multiple development stages,hosts and environments. The ability to transform from one stage to another and adapt to changing environments demands precise regulation of gene expression. Bioinformatic surveys of the sequenced genomes of T. gondii revealed a peculiar absence of DNA-binding transcription factors that are well-conserved from yeast through humans, but a wealth of epigenetic machinery present in T. gondii. Evidence from reports demonstrates that remodeling of the chromatin structure particularly through post-translational modifications of histones, such as acetylation,methylation, phosphorylation,ubiquitination,and sumoylation,is potentially a major process that coordinates regulation of its gene expression. In addition, no-coding RNAs may play an important role in modulating gene expression of T. gondii. These results provide reliable foundations for prevention of toxoplasmosis by revealing its pathogenic mechanism.
    The Signaling Systems in Echinococcus multilocularis
    NI Xing-wei,YAN Hong-bin,LOU Zhong-zi,JIA Wan-zhong*
    2012, 30(3):  15-233-237. 
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    Signaling pathway is the way by which cells receive various stimulation signals, and produce a series of corresponding responses, such as cell proliferation, differentiation and apoptosis. During infection with Echinococcus multilocularis, both parasite and host cells may secrete many cytokines such as insulin, which make stimulating signals transmitted into the cells through their receptors on the surface of cells. As a result, the parasite can grow and proliferate in the host. Study on related signaling pathways and their blocking drugs will play a crucial role in the control of alveolar echinococcosis caused by the larvae of E. multilocularis.
    Research Advances on Circumsporzoite Protein of Plasmodium
    HUANG Yan-ting, LU Xue-mei, JIN Xiao-bao, ZHU Jia-yong*
    2012, 30(3):  16-238-242. 
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    Circumsporzoite protein (CSP) is found in all the mature malaria parasites, which forms a dense coat on the sporozoite′s surface. CSPs contain approximately 400 amino acids and are organized into three domains: an N-terminal domain containing a conserved pentapeptide called region Ⅰ, a highly repetitive species-specific central domain, and a C-terminal domain containing another conserved sequence called region Ⅱ. It has been reported that the CSP fulfills vital roles in invading to the mosquito′s salivary glands, binding sporozoite to liver cells, and inactivating host cell protein synthesis machinery. Recently, researches pointed out that both of the vaccine and the targeted-drug-delivery-system based on CSP antigen reveal an immense prospect. This review presents a compilation of the protein at the molecular characterization, function and application level that have been described to date.
    Survey on a New Case of Visceral Leishmaniasis in Shanxi Province
    CAI Hui-xia1,2,ZUO Su-jun3,XU Wen-xing4,WU Wei-ping1 *,LI Jun1,XU Ke-jun5
    2012, 30(3):  17-188-190. 
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    A new case of visceral leishmaniasis (kala-azar) was reported by Changzhi CDC of Shanxi Province in September 2011. The case was investigated clinically and epidemiologically. The patient was a two-year-old boy who lived in Huangnian Town of Changzhi County in Shanxi Province. Clinical examination showed hepatosplenomegaly, consistent decrease of blood cells and Leishman-Donovan body in the bone marrow smear. The rK39 immune diagnosis test showed strongly positive. The case was diagnosed as kala-azar. After one course treatment of sodium stibogluconate, the patient's condition improved markedly. There were no cases of kala-azar in this region historically. Blood samples of 17 individuals and 5 domestic animals including 3 dogs were all negative in the rK39 immunodiagnostic test. It is speculated that the potential risk of kala-azar transmission exists in this region.
    Allele Genetype Analysis of Plasmodium vivax Merozoite Surface Protein1 (PvMSP-1) from Dangdong,Liaoning Province
    GENG Ying-zhi1, MAO Ling-ling1, TENG Cong1, AN Chun-li2, WANG Bo1, CHEN Jing-yi1, YAO Wen-qing1 *
    2012, 30(3):  18-242-244. 
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    Nested PCR method was used to amplify the Plasmodium vivax merozoite surface protein 1 (PvMSP-1) gene fragment containing the ICB5 and ICB6 region from Plasmodium vivax in Liaoning Province. The PCR products were digested by PvuⅡrestriction endonuclease and the digested fragments were observed by 1.5% agarose gel electrophoresis,and all followed by sequencing analysis and comparion. In 11 field isolates of P. vivax, two kinds of DNA fragments with 470 and 400 bp were produced respectively. After PvulI digestion,two Sal-1 type fragments (120 and 350 bp) were obtained from 5 samples of 470 bp. Single band of 400 bp appeared in 1 samples as Belem type. Two bands of 120 and 280 bp appeared from another 1 sample as recombination type III,and other 4 bands with 120 and 240 bp as Korean isolate. The principal types of PvMSP-1 alleles exist in malaria endemic areas in Liaoning Province with no mixed infection of two different type alleles.
    Killing Effect of Garlic Extract on Schistosoma japonicum cercariae and Oncomelania snails
    YANG Peng, DUAN Hong-yan, WANG Ce, SUN Jin-lian, LI Cheng-cheng, WANG Qing-li, SHEN Xiang-zhi, WANG Long, DONG Xing-qi, YANG Yi-mei*
    2012, 30(3):  19-245-246. 
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    The killing effect of different concentrations of garlic extract solution on Schistosoma japonicum cercariae and Oncomelania snails was observed under dissecting microscope. Mice were infected by cercariae through the abdominal skin daubed by garlic solution or by deionized water as control. The results showed that the cercariae were killed in (77.33±25.01) s in average, it needed (73.00±1.73)-(299.67±18.96) s under the garlic solution concentrations of 50.00%- 0.79% respectively, while the cercariae kept alive in 600 s in the control. The snails were killed in 1 d by 100% garlic solution but no death in the control in 2 d. No mouse daubed with different concentrations of garlic solution was found infected by schistosomes while 100% of the control mice got infected. It is concluded that the garlic shows satisfactory effect in killing cercariae and Oncomelania snails, and may prevent schistosome infection by daubing the skin.
    Pilot Observation on the Effect of Integrated Control Measures against Clonorchiasis in Pengjiang District of Jiangmen City
    ZHENG Nan-cai*,HUANG Bao-ming,LI Feng-ling,ZHOU Li-li,Liang Bai-nian,LUO Chun-hong,CHEN Xiao-jun,WU Er-ming,TAN Yan-fang
    2012, 30(3):  20-247-249. 
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    From June to December in 2008,five villages were randomly chosen from Pengjiang District of Jiangmen city and about five hundred residents from each village were examined for clonorchiasis by Kato-Katz method(three slides per specimen). Fifty residents from each village were re-examined one month after treatment. One year later 50 treated residents were chosen from Dalin village and Sanya village for fecal examination. Questionnairing was conducted to determine the knowledge rate on clonorchiasis prevention among residents. The percentage and usage of sanitary toilets were investigated. The average infection rate of clonorchiasis from five villages was 21.5%(537/2501). 86.6%(465/537)of clonorchiasis received treatment voluntarily. One month after treatment the infection rate in four villages declined significantly. The positive rate showed no significant difference between one month and one year after treatment in Dalin and Sanya villages(P>0.05). Questionairing indicated that 41.2%(170/413)of the clonorchiasis cases ate raw fish frequently, which was significantly higher than those non-infected people[4.2%,8/192](P<0.05). After health education, the knowledge awareness rate raised from 23.1%(135/584)to 84.5%(349/413)(P<0.05). The dissemination and usage of sanitary toilets were 93.2%(38 068/40 848)and 100%, respectively.
    mRNA Transcription of Translation Elongation Factor and Primase in Different Segments of Moniezia expansa
    WANG Zheng-rong, BO Xin-wen*, ZHAO Wen-juan, KANG Li-chao
    2012, 30(3):  21-250-252. 
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    A SYBR green real-time fluorescent quantitative PCR assay was developed for detection of Moniezia expansa mRNA with its beta-tubulin as an internal control. The results showed a good linear relationship (>0.99) between the Ct value and the concentration of positive plasmid for each gene from scolex and various proglottids. Real-time PCR showed that the expression abundance of translation elongation factor and primase was different. In conclusion, the transcription level of translation elongation factor and primase was high in both scolex and immature segment, suggesting that they may play a role in the development of scolex and immature segment.