Table of Content

30 December 2013, Volume 31 Issue 6

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Malaria Situation in the People’s Republic of China in 2012

XIA Zhi-Gui, FENG Jun, ZHOU Shui-Sen

2013, 31(6):  1-413-418. 

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Objective  To analyze malaria situation and epidemic characteristics in 2012 in China， and provide evidence and reference for National Malaria Elimination Programme（NMEP） performence.  Methods 　The epidemiological data of malaria cases reported through the annual malaria statistics reporting system in 2012 were collected and analyzed using Microsoft Excel 2010 and ArcGIS 10.0. 　Results　 Totally 2 718 malaria cases were reported from 620 counties of 31 Provinces/Municipalities/Autonomous Regions（P/M/A） in 2012, representing 39.3% reduction compared with 4 479 cases in 2011, and the annual incidence was 0.020 2/10 000. The cases were mainly reported from provinces of Yunnan（31.4%, 853/2 718）, Guangxi （8.1%, 220/2 718）, Jiangsu（7.3%, 198/2 718）, Hunan（5.8%, 158/2 718）, and Sichuan（5.7%, 155/2 718）. Among the 620 counties with reported cases, 8 counties including Motuo（8.1818/10 000） in Tibet, Ruili（8.248 9/10 000）, Yingjiang（3.021 4/10 000）, Longchuan（1.477 8/10 000）, Mangshi（1.4244/10 000）, Tengchong（3.1601/10 000）and Cangyuan（1.340 0/10 000） in Yunnan, and Shanglin（2.355 1/10 000） in Guangxi had an incidence between 1/10 000 and 10/10 000, 96 counties had an incidence between 0.1/10 000 and 1/10 000, and that of the others was below 0.1/10 000. The laboratory confirmed cases took 95.6%（2 599/2 718） while the other 4.4%（119/2 718）were clinically diagnosed. In detail, 39.7%（1 080/2 718） were P. vivax cases, 52.2%（1 419/2 718） were P. falciparum cases, 1.6%（44/2 718） were mixed infection of P. vivax and P. falciparum, and 2.1%（56/2 718） were P. ovale and P. malariae cases. However, the proportions of lab-confirmed cases in Xinjiang, Jilin, Heilongjiang, Tibet, Ningxia, Shanxi and Qinghai were below 75.0%. A total of 182（6.7%, 182/2 718） indigenous cases were reported from 41 counties in 5 provinces including 20 counties of Yunnan, 15 counties of Anhui, 4 counties of Hubei, 1 county of Tibet and 1 county of Guangxi, consisting of 38（20.9%, 38/182） clinically diagnosed cases（30 cases from Yunnan and 8 from Tibet）, 133（73.1%, 133/182）P. vivax cases（92 cases from Yunnan, 30 from Anhui, 9 from Hubei, 1 from Guangxi and 1 from Tibet）, and 9（4.9%, 9/182）P. falciparum cases as well as 2（1.1%, 2/182）mixed infections from Yunnan. The incidence of indigenous cases between 1/10 000 and 10/10 000 was found only in Motuo County of Tibet, and that of the others was below 1/10 000. Out of the 2 718 malaria cases, a proportion of 91.0% （2 474/2 718） were reported as the abroad-imported cases who distributed in 29 provinces, and the remaining 2.3%（62/2 718） were domestically-mobile cases reported from 10 provinces. Totally 145（5.3%, 145/2 718） severe cases were reported from 15 provinces and 15（0.6%, 15/2 718） malaria deaths were from 11 provinces.  Conclusion  Generally the indigenous malaria was reduced closer to the NMEP target, while malaria importation becomes an increasing challenge.

Ultrastructural Alterations of Adult Schistosoma japonicum Harbored  in Mice Treated with a Single Oral Dose of OZ78

XIAO Shu-Hua, SUN Jun, XUE Jian, DU Xi-Ling, ZHANG Hao-Bing

2013, 31(6):  2-419-427. 

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【Abstract】  Objective  To observe the ultrastructural alterations of adult Schistosoma japonicum induced by synthetic trioxolane OZ78.  Methods  Eight out of ten mice infected with 40-60 S. japonicum cercariae for 35 d were treated orally with OZ78 at a single dose of 400 mg/kg. Four groups of two mice were killed at 24 h, 3 d, 7 d, and 14 d post treatment, and schistosomes were recovered by perfusion technique, fixed, and examined by transmission electron microscopy. Schistosomes obtained from the remaining two untreated mice served as control.  Results  After infected mice were treated with OZ78 for 24 h, the prominent alterations in tegument of both male and female worms were observed, which revealed in flattened surface due to swelling of cytoplasmic processes, irregular expansion in distal end of cytoplasmic processes accompanied by decrease in rod-like and discoid-like secretory bodies, focal lysis of tegumental matrix; fusion of some cytoplasmic processes to form a large piece, disruption or disappearance of basal membrane, and destruction of internal structures in sensory organelles. In the subtegument, no or slight swelling and focal lysis of muscle bundles were seen, while the syncytium beneath the muscle showed enlargement of nucleus with indistinction of partial nuclear membrane, formation of small vacuoles due to focal lysis of chromatin, and emergence of degenerated mitochondria in perinuclear cytoplasm. As to parenchymal tissues, the major alterations included degeneration of mitochondria, formation of some small vacuoles and myelin-like structures. In gut epithelial cells, the prominent alterations were irregular enlargement of nucleus with light lysis of nucleoli and fusion of partial bi-layer nuclear membrane, degeneration of mitochondria in cytoplasm and collapse of microvilli. At this time point, in the vitelline cells of female worms, the most significant alteration was the collapse of many vitelline droplets, which led to release of the vitelline balls, followed by their lysis and fusion. Three to 7 d post treatment, the damage to the worms aggravated either in extent or in severity along with time. The significant damages to male and female worms were fusion of cytoplasmic processes, peeling or collapse of damaged cytoplasmic processes resulting in exposure of muscle bundles, severe destruction of sensory organelles and syncytium, focal or extensive swelling and lysis of muscle bundles, emergence of some larger piece of degenerated parenchymal tissues and severe damage to the gut epithelial cell. While in the vitelline cells of female worms, decrease in the number of vitelline droplet, focal lysis of nucleus and extensive lysis of parenchymal tissues among the vitelline cells were also observed. Fourteen days post OZ78 dosing, male and female worms which survived the treatment showed some renovation in damaged tegument and subtegument, while most gut epithelial cells and vitelline cells still revealed in prominent injury.  Conclusion  The results demonstrate that OZ78 possesses an extensive damage to the ultrastructure in tegument and subtegument tissues including syncytium, parenchymal tissues, gut epithelial cells, and vitelline cells of adult S. japonicum.

Surveilance and Response for Schistosomiasis Japonica Based on Sentinel Mice Examination for Cercariae-infested Water in Risk Region, 2012

ZHENG Hao, LI Shi-Zhu, CAO Chun-Li, ZHANG Li-Juan, SUN Le-Ping, YANG Kun, TU Zu-Wu, LI Yi-Yi, YANG Wei-ping, GU Xiao-Nan, WU Zi-Song, FENG Xi-Guang, ZHU Rong, XU Jing, XIAO Ning, ZHOU Xiao-Nong

2013, 31(6):  3-428-432. 

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Objective  To monitor water body infestation in schistosomiasis high-risk areas with sentinel mouse technique.  Methods  A total of 72 surveillance sites from 47 counties were selected in Hunan, Hubei, Jiangxi, Anhui, Jiangsu, Yunnan, and Sichuan. The water infectivity of Schistosoma japonicum was determined in the surveillance sites by using sentinel mice during June-July and September, 2012.  Results  Among the 3 283 sentinel mice which were placed in 72 sites, 3 062 （93.3%） were recovered and dissected. Infected sentinel mice were found in six sites accounting for 8.3% （6/72） of the total surveillance sites, with an occurrence rate of sites with infected mice of 8.3% in June-July and 2.8% in September. 33 infected mice were discovered with a total infection rate of 1.08% （33/3 062）. 1 085 adult worms were collected, with a mean worm burden of 32.9 worms per mouse in infected sentinel mice. 4 positive sites were in Hunan and 2 were in Jiangxi. Local acute schistosomiasis or suspected local acute cases which detected elsewhere were reported in 2 positive sites. Some follow-up activities were conducted in the 6 positive sites.  Conclusion　 Compared with those in 2010, the schistosomiasis risk areas are shrinking in 2012. However, some regions are still the schistosomiasis high-risk areas.

Establishment of Policy Indicators of Adaptation to the Impact of Climate Change on the Transmission of Schistosomiasis and Malaria in China

QIAN Ying-Jun, LI Shi-Zhu, XU Jun-Fang, ZHANG Li, FU Qing, ZHOU Xiao-Nong

2013, 31(6):  4-433-437. 

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Objective  To set up a framework of indicators for schistosomiasis and malaria to guide the formulation and evaluation of vector-borne disease control policies focusing on adaptation to the negative impact of climate change. 　Methods　 A 2-level indicator framework was set up on the basis of literature review, and Delphi method was applied to a total of 22 and 19 experts working on schistosomiasis and malaria, respectively. The result was analyzed to calculate the weight of various indicators. 　Results 　A total of 41 questionnaires was delivered， and 38 with valid response(92.7%). The system included 4 indicators at first level，　i.e. surveillance， scientific research， disease control and intervention, and adaptation capacity building, with 25 indicators for schistosomiasis and 21 for malaria at the second level. Among indicators at the first level, disease surveillance ranked first with a weight of 0.32. Among the indicators at the second level， vector monitoring scored the highest in terms of both schistosomiasis and malaria. 　Conclusion  The indicators set up by Delphi method are practical，universal and effective ones using in the field， which is also useful to technically support the establishment of adaptation to climate change in the field of public health.

Serodiagnosis of the Recombinant Multi-epitope Antigens from Antigen B Subunits of Echinococcus granulosus

JIANG Li, FENG Zheng, ZHANG Yao-Guang, WANG Zhen-Yu

2013, 31(6):  5-438-442. 

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Objective  To evaluate the diagnostic value of six recombinant multi-epitope antigens and three AgB subunit antigens from antigen B subunit of Echinococcus granulosus.  Methods  A linker sequence was inserted into the sequence of MEA-26 to make it an MEA-49. I-TASSER on-line server was used to analyze protein structure. The reactivity of two multi-epitope recombinant antigens with the same target sequence but in different tertiary structure was compared. The reactivity of six multi-epitope antigens（MEA-8, MEA-20, MEA-26, MEA-36, MEA-49, and MEA-52）, 3 subunit antigens （AgB1, AgB2, and AgB4）, and 2 control antigens （Trx and linker） was determined by indirect ELISA. The assays were performed on 232 serum samples separated as follows: 112 sera from patients with cystic echinococcosis, 35 sera from individuals with alveolar echinococcosis, 43 sera from patients with cysticercosis and 42 sera from healthy individuals. Their diagnostic performance was assessed by receiver operating characteristic （ROC） curve analysis.  Results  Tertiary structure prediction showed that the epitope regions of MEA-26 were closer to each other and aligned in parallel, while that in MEA-49 were farther apart from each other and formed two independent domains. Serological analysis revealed that the mean P/N value （2.88±2.02）, sensitivity （92%） and diagnostic efficiency （89%） of MEA-49 were higher than that of MEA-26 （2.54±2.02, 78% and 82%）. MEA-20　（2.24±1.31）, MEA-26 （2.54±2.02）, MEA-36 （2.44±1.51）, MEA-49（2.88±2.02） and MEA-52（2.50±1.37） showed a high reactivity to the sera from patients with cystic echinococcosis, which was superior to that of AgB1（2.15±1.26）. There was no significant difference in the reactivity to sera from individuals with alveolar echinococcosis between multi-epitope antigens and AgB1（P>0.05）. MEA-52 showed a high diagnostic sensitivity in cysticercosis cases（1.27±0.70）, superior to that of AgB1 （0.95±0.13） （P<0.01）. The reactivity of MEA-8 （1.04±0.15） to sera from healthy individuals was significantly higher than that of AgB1 （0.89±0.07） （P<0.01）. ROC analysis showed in the cases of cystic echinococcosis, the diagnostic sensitivity accomplished with AgB1, AgB2, and AgB4 was 77%, 55%, and 66%, respectively; the multi-epitope antigens of MEA-49（92%）, MEA-36（92%）, MEA-52 （87%）, and MEA-26（78%） revealed a higher sensitivity than AgB1.  Conclusion  The reactivity of multi-epitope antigens is superior to that of AgB subunit antigens. The reactivity of MEA-49 is higher than that of MEA-26 which has the same target sequence but in different tertiary structure.

Histological Observation of the Central Nervous System in Simulium （Wilhelmia）xingyiense（Diptera ∶ Simuliidae）

XUN Hui, DING Kai-Ze, YANG Ming, CHEN Han-Bin

2013, 31(6):  6-443-447. 

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Objective  To investigate the morphological characters of the central nervous system in mature larvae， mature pupae and newly emerged adults of Simulium (Wilhelmia) xingyiense.  Methods  From August to November 2009, the blackfly larvae were collected from the rivulets nearby Niujiao Island in Huaxi of Guiyang City. The mature larvae of S. (Wi.) xingyiense were confirmed based on the diagnostic characteristics of gill spots, postgenal cleft, and rectal gill. The mature pupae were obtained from the rivulets of Da′ao Town and Qingyan Town in Guiyang in March 2011, which were selected according to the characteristics of cocoon and respiratory filaments. Nervous system of the larvae， pupae and newly emerged adults was observed under the light microscope with HE-stained paraffin sections.  Results  The central nervous system was composed of brain, subesophageal ganglion and ventral nerve cord. The brain of the larva was divided into two narrowly interconnected egg-shaped lobes. Ventral nerve-cord of the larva consisted of three pairs of thoracic ganglia and eight pairs of abdominal ganglia. The brain of the pupa and adult was composed of protocerebrum, deutocerebrum, and tritocerebrum. The ventral nerve cord of the pupa and adult was similar to that of larva. From outside to inside, the structure characters of the brain and ganglia were similar with nerve sheath, neurocyte and neuropile. The neuropile of protocerebrum contained a pair of mushroom bodies, a central complex and a pair of accessory lobes. The optic lobe was composed of medulla interna, medulla externa and lamina ganglionaris. The deutocerebrum consisted of the antennal lobe and the dorsal lobe. The tritocerebrum connected to the subesophageal ganglion by perioperative esophageal nerve.  Conclusion  The central nervous system of S. (Wi.) xingyiense is similar to other simuliid blackflies. There is a difference in the number of abdominal ganglion of the mature larva.

Effects of Anti-osteopontin Antibody on Expression of MMP-2 and TGF-β1 in Hepatic Alveolar Hydatid Tissue of Gerbil

LI Tong, WU Xiang-Wei, ZHANG Yong-Guo, GAO Liang-Liang, CAO Yu-Wen, SUN Hong, ZHANG Shi-Jie

2013, 31(6):  7-450-453. 

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Objective  To observe the effect of anti-osteopontin antibody on the level of matrix metalloproteinase （MMP-2） and TGF-β1 in gerbils infected with Echinococcus multilocularis.  Methods  One hundred and eighty gerbils were infected with echinococcus protoscoleces（approximately 400 for each gerbil） by abdominal opening inoculation in liver. The gerbils were randomly divided into three groups: anti-osteopontin antibody experiment group（group A）, rabbit serum injection group（group B）, and model group（group C）. Gerbils in groups A and B were injected with anti-osteopontin antibodies and rabbit serum （0.15 ml/gerbil） via tail vein, respectively. Ten gerbils from each group were sacrificed at 20, 60, 100, 140, 180, and 220 days post-infection, respectively. The liver tissue with hydatid cysts were collected and the expression of MMP-2 and TGF-β1 was observed by immunohistochemistry staining（SP method）.  Results  E. multilocularis hydatid tissue spreaded over the liver and abdominal cavity. There was no significant difference in the number of MMP-2-positive gerbils among the three groups（P＞0.05）. At 100, 140, and 180 days post-infection, the number of TGF-β1-positive gerbils in group A （3, 2, and 2） was considerably less than that of group B （8, 8, and 9） and group C （8, 9, and 9）（P<0.05）.  Conclusion  Anti-osteopontin antibody can reduce the expression of TGF-β1 in hepatic alveolar hydatid tissue of gerbils at certain time, but have no effect on MMP2.

Proteomic Study of the Hippocampus Tissue from Rats with Chronic Toxoplasma gondii Infection

ZHOU Yong-Hua, FAN Hong-Jie, ZHANG Ying, HUANG Yu-Zheng, XU Yong-Liang, TAO Yong-Hui, GAO Qi

2013, 31(6):  8-454-457. 

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 Objective  To observe the proteome changes in the hippocampus tissue of rats with chronic Toxoplasma gondii infection.  Methods  Six male SD rats were randomly divided into control group and infection group. Each rat in infection group was intraperitoneally injected with 4×107 purified T. gondii tachyzoites. Rats in the control group received equivalent volumes of sterile normal saline. At the fifth day post-infection, blood samples were taken from the lateral tail vein and Giemsa staining of blood cells was performed to find Toxoplasma gondii. Rats were dissected at the 10th week post-infection, total protein in the hippocampus was separated by using two-dimensional gel electrophoresis（2-DE）. After Coomassie blue staining, the Image Analysis software was used to select and separate proteins on the gel. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry（MALDI-TOF-MS） was used for peptide mass fingerprint （PMF）. Proteins were identified by using Mascot software to search the MSDB and SwissProt databases.  Results  Microscopy examination of blood smears confirmed that the rats in infection group were all infected by T. gondii. The number of protein spots of rats from infection group and control group was 311±19 and 327±13, respectively. Compared with the control group, 5 protein spots disappeared, 4 protein spots were up-regulated and 7 were down-regulated in the infection group. The 9 differentially expressed protein spots were identified by MALDL-TOF-MS: phosphoglycerate kinase 1, similar to alpha-enolase, glutamine synthetase, creatine kinase, creatine kinase B-type, ATP synthase, aconitase 2, mitochondrial precursor, actin and an unnamed protein. The first three proteins were up-regulated and the other five proteins were down-regulated in infection group. 　Conclusion  Nine differential expression proteins are found from the hippocampus tissue in rats chronically infected with T. gondii and normal SD rats.

Prevalence and Control of Schistosomiasis in Jiangxi Province during 2002-2012

CHEN Hong-Gen, GU Xiao-Nan, ZENG Xiao-Jun, LIN Dan-Dan, HANG Chun-Qin, LV Chang-Biao, CHEN Zhe, LI Zhao-Jun

2013, 31(6):  9-458-463. 

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Objective  To analyze the status of prevalence and control of schistosomiasis in Jiangxi Province from 2002 to 2012.  Methods  Endemic status and control effectiveness during the period from 2002 to 2012 was analyzed by using a combination of field survey, data collection and retrospective investigation.  Results  From 2002 to 2012, schistosomiasis control made significant achievements in Jiangxi Province. The number of counties which have reached the criteria of transmission interruption of schistosomiasis increased from 19 in 2002 to 22 in 2012. The number of patients with schistosome infection decreased from 128 331 in 2002 to 73 102 in 2012. The positive rate of serological test and fecal examination in residents decreased from 12.2% and 4.2% in 2002 to 5.2% and 0.4% in 2012, respectively. Acute infection reduced from 146 cases in 2002 to 3 cases in 2012, but advanced cases increased by 42.4%. The positive rate of fecal examination in cattle reduced from 4.7% in 2002 to 1.3% in 2012. The density of living snails and infected snails significantly decreased to 0.083 5/0.1 m2 and 0.000 037/0.1 m2 in 2012, respectively. However, the snail-ridden areas slightly increased. Jiangxi Province reached the criteria of endemic control for schistosomiasis in 2008. During 2002-2012, control activities were intensified. Examination and chemotherapy for human increased by 65.2% and 65.5%, while 23.4% and 251.1% for cattle. Mean while, area with snail control activities increased by 617.8%. An integrated strategy of infection source control was implememted in the Province.　 Conclusion  During 2002 to 2012, endemic index of schistosomiasis in Jiangxi showed a steady decline after 2005. Currently, schistosomiasis prevalence shows a low level. The endemic indicators of infection in human and livestock as well as Oncomelania snails have been kept stable at low level.

Prevalence of Angiostrongylus cantonensis Infection in Snails for Sale in Fuzhou and Xiamen

LI Li-Sha, ZHANG Rong-Yan, FANG Yan-Yan, OUYANG Rong, XIE Han-Guo, JIANG Dian-Wei, XIE Xian-Liang, CHEN Zhu-Yun, ZHENG Guo-Bin

2013, 31(6):  10-464-466. 

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Objective  To investigate the prevalence of Angiostrongylus cantonensis infection in snails for sale from Fuzhou and Xiamen of Fujian Province.  Methods  During 2009-2012, two markets and five restaurants from each city were selected as surveillance sites. A. cantonensis infection rate in Pomacea canaliculata, Bellamya aeruginosa and Cipangopaludina cathayensis was examined two times per month. More than 50 P. canaliculata or C. cathayensis, and about 500 g B. aeruginosa were collected in each site. A. cantonensis larvae infection was determined by lung-microscopy in P. canaliculata, and by tissue homogenate method in C. cathayensis and B. aeruginosa, respectively.  Results  In markets, a total of 5 744 P. canaliculata were collected, and the infection rate of A. cantonensis larvae was 13.8% （753/5 744） with the lowest prevalence in 2009 （8.4%, 28/334） and the highest one in 2011 （16.7%, 361/2　160）. The overall infection rate of A. cantonensis showed an increasing trend over the past years （P＜0.05）. In restaurants, 879 P. canaliculata snails were examined in Xiamen City, and the infection rate was 12.8% （877/6 879）. No significant difference was found among years （P＞0.05）. A. cantonensis larvae were found from P. canaliculata for sale in different seasons with no statistical difference （P＞0.05）. 19 843 B. aeruginosa snails were collected in markets from the two cities, and the infection rate was 0.2% （31/19 843）. The infection rate was highest in 2011 （0.3%, 16/5 953） and lowest in 2010 （0.04%, 2/4 706）. All the 361 C. cathayensis snails were negative.  Conclusion  A. cantonensis larvae are found in P. canaliculata and B. aeruginosas from markets and restaurants of Fuzhou and Xiamen in different seasons during the years.

Analysis on Theses of the Chinese Journal of Parasitology and Parasitic Diseases in 2009-2012

YI Feng-Yun, QU Lin-Ping, YAN He, SHENG Hui-Feng

2013, 31(6):  11-467-470. 

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The published articles at the Chinese Journal of Parasitology and Parasitic Diseases in 2009-2012 were statistically analyzed. Among 547 papers published in the four years， original articles occupied 45.3%（248/547）. The number of authors was 2 712， with an average cooperation degree of 5.0, and the co-authorship accounted for 95.4% of the papers. Authors were mainly from colleges/universities （51.9%, 284/547）, institutions for disease control （34.4%, 188/547） and hospitals health centers（13.7%, 75/547）. The average publishing delay was 212, 141, 191 and 207 d in 2009-2012. Statistical analysis reflected the characteristics and academic level for improving the quality of the journal, and revealed the latest development and trends.

Research Progress on Polymorphism in the Merozoite Surface Protein-3α Gene of Plasmodium Vivax

XU Chao, XU Xiu-Lai, HUANG Bing-Cheng

2013, 31(6):  12-473-476. 

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The merozoite surface protein-3α of Plasmodium vivax（PvMSP-3α） is a surface protein that is expressed during the asexual blood stages. With a high polymorphism,  PvMSP-3α gene has been used as a suitable epidemiology and genotype marker. Moreover, it might be an important malaria vaccine candidate. This paper summarizes the research progress on the molecular structure, gene polymorphism and genotypes of PvMSP-3α.

Cloning and Bioinformatics Analysis of Rhoptry Protein 11 of Toxoplasma gondii

ZHANG Xiao-Lei, ZHANG Jin-Shun, JIA Xiao-Hui, XU Yun-Peng, ZHANG Ying, WANG Chun-Miao, WANG Yan, LU Zhi-Min, ZHAO Jian-Ling, JIA Tian-Jun

2013, 31(6):  13-447-449. 

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Total RNA was extracted from tachyzoites of RH strain of Toxoplasma gondii. The open reading frame of ROP11 gene was amplified by using a pair of specific primers designed according to the coding sequence of ROP11 gene （Accession No. DQ077905）. The RT-PCR product was digested by restriction enzyme EcoRⅠ and NotⅠ, and then ligated into a pGEX-6P-2 vector. The recombinant plasmid was transferred into E. coli XL-Blue. The positive clones was selected by colony PCR, and confirmed by the double restriction enzyme digestion and sequencing. The RT-PCR product was 1 548 bp. The recombinant plasmid was confirmed by colony PCR and double restriction enzyme digestion. Sequencing results showed that the obtained ROP11 gene was 1 548 bp （Accession No. KC456639）. There was a high sequence consistency （99%） between the obtained ROP11 gene sequence and the Toxoplasma ROP11 gene from GenBank. Bioinformatics analysis showed that the ROP11 protein （Mr 57 020） consisted of the signal peptide （amino acids 1-26）, 12 conservative domains, a serine/threonine protein kinase catalytic domain （amino acids 170-511）, and two potential N-glycosylation sites.

Anti-tumor Effect of the Whole Worm Extract of Ascaris lumbricoides on Lewis Lung Carcinoma in Mice

YANG Xiao-Jun, YANG Jun-Ping, HUANG Yan-Qin, LIANG Hua, YUAN Keng

2013, 31(6):  14-470-472. 

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Forty-five C57BL/6 mice were randomly divided into five groups（A-E）. Group B and D served as the control group of A and C. Each mouse of group A was intraperitoneally injected with 0.1 ml whole worm extract of Ascaris lumbricoides every other day, and 10 days later injected with 0.1 ml Lewis lung carcinoma（LLC） cells at right axillary subcutaneously region. Mice of group B were injected with normal saline and then developed tumor model. Each mouse of group C was injected with 0.1 ml LLC cells, and two days later, injected with 0.1 ml whole worm extract of A.lumbricoides every other day for 5 times. After the tumor model developed, mice in group D were injected with normal saline. Group E was the negative control group. Time intervals between implantation and active growth and tumor weight were recorded. Tumor inhibition rate was calculated. The average time interval between tumor implantation and measurable tumor growth for groups A, B, C and D was （7.0±1.1）, （6.0±0.7）, （9.0±1.2） and （7.0±0.9） days. Tumor weight of group A [（722.2±413.5） mg] was heavier than that of group B [（338.9±282.2） mg] （P＜0.05）. The tumor inhibition rate was the highest in group C （33.3%）. Tumor weight of group C [（237.8±101.8） mg] was lighter than that of group D [（356.7±176.9） mg] （P＜0.05）. The results indicated that the tumor formation is affected by the whole worm extract of A. lumbricoides which may have an inhibitory effect on tumour growth.

Therapeutic Efficacy of Allitridin and Azithromycin on Cryptosporidium Infection in Mice

WANG Dong, ZHANG Yuan-Yuan

2013, 31(6):  15-477-479. 

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Seventy Kunming mice were randomly divided into four groups named as normal control group（10）, Cryptosporidum infected group（20）, allitridin treatment group（20） and azithromycin treatment group（20）. After infection, each mouse in allitridin treatment and azithromycin treatment groups were given 1.5 mg/d allitridin or 2 mg/d azithromycin by gavage for 12 days, respectively. The mice were observed and the oocysts shedding in fecal pellets were counted daily after treatment. The level of secretory IgA（sIgA） in intestinal fluid of proximal jejunum was determined. The pathological changes in mucous membrane of proximal jejunum were observed by HE staining. Compared with infection group, the level of oocyst shedding was considerably lower in the two treatment groups（P＜0.05）. There was a significant difference in sIgA level in intestinal fluid between allitridin treatment group（5.16 μl/ml） and azithromycin treatment group（3.7 μl/ml）（P＜0.05）. Pathological examination of intestinal mucous membrane showed that swollen mucous membrane and focal necrosis of tissue were observed in infection group, and less inflammatory cell infiltration occurred in the two treatment groups. Allitridin and azithromycin are effective in the treatment of Cryptosporidium infection. Allitridin can enhance the defense function of intestinal mucosa.

Cloning，Expression and Analysis of Der f Mag 29 Allergen of Dermatophagoides farinae

SUN Jin-Xia, SHU Li-Li, ZHOU Ying, BIAN Yong-Hua, YANG Li, WANG Nan, CUI Yu-Bao

2013, 31(6):  16-480-482. 

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The full-length Mag 29 gene of Dermatophagoides farinae was amplified by RT-PCR with a pair of specific primers. The PCR product was cloned into pCold TF DNA vector. The constructed plasmid pCold TF-Mag 29 was transformed into E.　coli BL21 and followed by expression of the protein induced by IPTG. The recombinant protein was analyzed by SDS-PAGE. The full-length Mag 29 gene was 429 bp. A specific band （M_r 63 000） were detected in the whole cells, the supernatant, and the precipitate. Bioinformatics analysis revealed that Mag 29 protein was composed with 142 amino acid residues with a calculated molecular weight of M_r 15 100, and its secondary structure was composed of alpha helix （55.63%）, extended strand（3.52%）, and random coil （40.85%）. The Mag 29 allergen was a hydrophilic and cytoplasmic protein, and shared a high degree homology with the heat shock protein 70 family.

Polymorphism Analysis on the Genotypes of Circumsporozoite Protein of Plasmodium vivax with PRC-RFLP

LIU Ying, ZHOU Rui-Min, CHEN Wei-Qi, QIAN Dan, ZHAO Yu-Ling, ZHAO Xu-Dong, XU Bian-Li, SU Yun-Pu, ZHANG Hong-Wei

2013, 31(6):  17-483-485. 

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Thirty-seven blood samples from Plasmodium vivax-infected patients were collected in Henan Province in 2011. The gene for circumsporozoite protein（PvCSP） was amplified by nested PCR. According to the epidemiological data, among the 37 Plasmodium vivax malaria cases, 26 were indigenous cases and 11 were imported cases. A fragment of 750 bp was obtained in the 37 blood samples. The 750 bp PCR product was digested with restriction endonuclease Alu-Ⅰ and Mva-Ⅰ, and genetic polymorphism was investigated with PCR-RFLP. PCR-RFLP analysis of PvCSP revealed that the parasites were PV-Ⅰtype（97.3%, 36/37） and PV-Ⅱ type（2.7%, 1/37）. 26 samples from indigenous cases were exclusively PV-Ⅰ type with PV-Ⅰ-b type（57.7%, 15/26） and PV-Ⅰ-a type（42.3%, 11/26）. Among 11 samples from imported cases, PV-Ⅰ-a type accounted for 90.9% （10/11）, and 1 PV-Ⅱ type accounted for 9.1% （1/11）, no PV-Ⅰ-b type were detected.

Evaluation of Transfection Effectiveness Using Fluorescein-labelled Oligonucleotides and Entraster-R siRNA Transfection into Plasmodium falciparum

ZHOU Hong-Chang, GAO Yu-Hui, SHAO Sheng-Wen, ZHANG Hui, ZHANG Ting

2013, 31(6):  18-487-489. 

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 The cultured Plasmodium falciparum parasites were synchronized twice by 5% sorbitol treatment twice（8-hour window）, and then incubated at 37 ℃ for 16 h. Parasites were transfected with fluorescein-labelled oligonucleotides （group A） or fluorescein-labelled oligonucleotides+Entranster-R siRNA transfection reagent （group B）. After 5 h a part of parasites was evaluated by fluorescence microscopy and flow cytometry. The rest of parasites were washed with RPMI 1640 medium, and then incubated with 500 μl new medium containing 2% fresh erythrocytes for another 12 h, and detected by flow cytometry. The fluorescein-labelled oligonucleotides were localized in erythrocytes in group B, but nearly no fluorescence was observed for group A. Flow cytometry analysis indicated that the transfection efficiency of group B [（47.40±3.39）%] was higher than that of group A [（0.60±0.27）%]. In the second cell cycle, the transfection efficiency in group B was （26.85±2.90）%, while that of group A was nearly zero. The results indicated that Entranster-R siRNA transfection reagent may increase the oligonucleotides transfection efficiciency.

Ultrastructure of the Digestive System in Dermatophagoides farinae （Acariformes  ∶  Pyroglyphidae）

WANG Yue-Meng, LIU Xiao-Yu, JIANG Cong-Li, HUANG Li-Nian, SUN Xin, LIU Zhi-Gang

2013, 31(6):  19-490-492. 

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Fifty living mites(Dermatophagoides farinae) were fixed in 2.5% glutaraldehyde, postfixed in 1% osmium tetroxide, dehydrated in a graded ethanol series, embedded in embedding medium. The ultrastructure of the digestive tract in D. farinae was observed by serial ultrathin sections with a transmission electron microscope. The alimentary canal of D. farinae consists of the cuticle-lined foregut and hindgut separated by a microvilli-lined midgut (anterior midgut, posterior midgut). There are different types of epithelial cells in the anterior midgut. The microvilli of epithelial cells in posterior midgut are longer than that of the anterior midgut. In posterior midgut, the food bolus is surrounded by the peritrophic membrane. The midgut is the main site of digestion and absorption.