Table of Content

30 October 2012, Volume 30 Issue 5

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Cloning， Expression and Function Analysis of Thioredoxin-1 Protein of Schistosoma japonicum

LIU Jian 1，XU Bin 2，ZHANG Xiao 1，2，LIU Lu 1，2，HU Wei 2，3，WANG Xiao-ningLIU Jian 1，XU Bin 2，ZHANG Xiao 1，2，LIU Lu 1，2，HU Wei 2，3，WANG Xiao-ning 1 *

2012, 30(5):  1-335-340. 

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【Abstract】   Objective   To clone and express recombinant thioredoxin-1 protein of Schistosoma japonicum (SjTrx-1）， determine its enzyme activity， analyze the stage-specific transcription of SjTrx-1 gene and observe the distribution of SjTrx-1 protein in adult worm.  Methods  SjTrx-1 coding sequence identified from S. japonicum cDNA library was amplified by PCR. Subsequently， the obtained target fragment was subcloned into pET28a， and the recombinant plasmid was then transformed into E. coli BL21 (DE3） cells and induced with IPTG for protein expression. The recombinant SjTrx-1 protein was purified with Ni-NTA resin and the enzyme activity was determined by DTT/insulin reduction method. The rabbit antiserum was prepared by immunizing New Zealand white rabbit with purified recombinant SjTrx-1 protein. Indirect immunofluorescence staining with the prepared polyclonal antibody was used to investigate the distribution of SjTrx-1 protein in adult worm. Total RNA was extracted from different stages of S. japonicum and reverse-transcribed into cDNA samples. RT-PCR was used to analyze the SjTrx-1 transcription level of each development stages.  Results  The expression vector of SjTrx-1/pET28a was established and the recombinant SjTrx-1 protein was expressed as soluble form in E. coli (about Mr 38 000）. The purified SjTrx-1 protein was obtained through Ni-NTA affinity purification and was demonstrated with adequate enzyme activity in vitro. The stage-specific analysis showed that SjTrx-1 was expressed in each of the developmental stages studied， with increased level in sporocysts， schistosomula and adult worms， and relatively low level in miracidia and cercaria. Immunolocalization revealed SjTrx-1 was widely spread in the worm without tissue-specific.  Conclusion  SjTrx-1 is an abundant and widely distributed protein with increased level in sporocysts， schistosomula and adult worms.

Identification of Glycosylphosphatidylinositol-anchored Protein from Schistosoma japonicum

CAO Qin-yan，XUE Yan-feng，SHEN Li *

2012, 30(5):  2-343-348. 

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【Abstract】   Objective   To identify glycosylphosphatidylinositol （GPI） anchored protein of Schistosoma japonicum.  Methods  Based on the gene sequence of Schistosoma mansoni GPI anchored protein Sm200（GenBank Assess No：XM_002569560.1）， bioinformatics analysis was performed to find out its homologous gene sequence in S. japonicum， then a selected partial coding sequence （SjGPIs， about 933 bp） from the homologous gene sequence were amplified， and cloned into PET-28a（+） vector. The recombinant plasmid pET-28a（+）-SjGPIs were transformed into E. coli Top10 cells and induced with IPTG for protein expression. The recombinant protein SjGPIs was purified with Ni-NTA resin， and the purified recombinant SjGPIs protein was used as antigen to prepare antiserum in New Zealand rabbit. The antiserum was used to detect S. japonicum GPI-anchored protein. To identify a GPI-anchored protein， the detected protein were identified by phosphatidylinositol-specific phospholipase C （PI-PLC） digestion. White blood cells from S. japonicum-infected mice was examined whether they endocytosed GPI-anchored proteins by Western blotting.  Results   The homologous gene sequence of S. mansoni GPI Sm200 gene was found in S. japonicum genome. A 3 495 bp coding sequence was obtained， containing the complete C-terminal sequence. The selected gene sequence （SjGPIs） were amplified and the recombinant plasmid pET-28a（+）-SjGPIs was established. According to the analysis of C-terminal sequence， Western blotting and enzyme digestion of PI-PLC， a GPI-anchored protein was present in S. japonicum tegument （about Mr 200 000）， named SjGPI200. The protein was detected in white blood cells of infected mice.  Conclusion  SjGPI200 protein exists in S. japonicum， and anchored to parasite tegument via GPI.

Study of Tacrolimus Intranasal Treatment for Allergic Asthma in Mice

YU Hai-qiong1，2，YUAN Ping2，HUANG Ying-hua1，LI Hui-qiong1，ZHOU Yi-ping1 *

2012, 30(5):  3-349-353. 

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【Abstract】   Objective   To explore the efficacy of intranasal treatment by immunosuppressant tacrolimus for allergic asthma and its mechanism in mice.  Methods  24 female BALB/c mice were randomly divided into 4 groups: group A （negative control）， group B （model control）， group C （low dose treatment）， and group D （high dose treatment）. Mice in group A were treated with saline （100 μl）. Other groups were sensitized intraperitoneally with allergen extracts of Dermatophagoides farinae （Der f） absorbed to Al（OH）3 at day 0， 7， and 14. From day 28， groups A， B， C， and D were intranasally treated with saline， PBS， 0.01% tacrolimus， and 0.1% tacrolimus， respectively， once per day for 7 d， and followed by intranasal challenge with 50 μl Der f extracts in the mean time. 24 h after the last challenge， the airway hyper-responsiveness （AHR） were detected. At 48 h after the last challenge, the mice were sacrificed, the bronchoalveolar lavage fluid （BALF） was collected, the lungs and spleen were aseptically removed. The total cell number and cell classification of BALF were recorded. The level of interleukin-4 （IL-4）， interleukin-5 （IL-5）， interferon-γ （IFN-γ） in BALF and in spleen cells culture supernatants was detected by ELISA. The lung inflammation and mucus secretion were observed in mice by HE （haematoxylin and eosin） staining and AB （Alcian Blue） staining.  Results  Compared with group B， AHR （P<0.05） and airway inflammation in group D significantly reduced. The number of total cells ［（29.92±5.20）×104/ml］ （P＜0.05） and eosinophils ［（4.33±0.75）×104/ml］ （P＜0.01） in group D greatly decreased than those of group B ［（59.33±5.99）×104/ml and （22.67±5.65）×104/ml］. The level of IL-4 ［（22.49±4.96） pg/ml］ （P＜0.05）, IL-5 [（43.90±13.15） pg/ml］ （P＜0.01） and IFN-γ ［（10.17±1.09） pg/ml］ （P＜0.05） in BALF significantly decreased （P<0.05） than those of group B ［（57.02±7.38）， （133.49±15.63） and （15.32±3.23） pg/ml, respectively］. The level of IL-4 ［（22.54±4.58） pg/ml］， IL-5 ［（36.31±20.85） pg/ml］ and IFN-γ ［（11.28±1.79） pg/ml］ in spleen cell culture supernatant all sig-nificantly decreased （P<0.05） than those of group B ［（56.34±6.21）， （72.32±6.23） and （18.82±1.88） pg/ml, respectively］. There was no significant difference between group C and group B.  Conclusion  Tacrolimus shows certain immune therapeu-tic effect on dust mite sensitized mice， and this effect may be attributed to its inhibition on T lymphocyte factor secretion.

Study on Immune Response in ICR Mice by Immunization with Recombinant Myophilin Vaccine against Echinococcus granulosus

MA Rui1,2, SHI Zhi-yun1, WANG Ya-na2, LI Zong-ji2, SUN Jun-feng2，ZHAO Wei1 *

2012, 30(5):  4-357-360. 

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【Abstract】   Objective   To investigate the immune response and the protection in mice induced by the recombi-nant myophilin protein of Echinococcus granulosus.  Methods  Thirty-six male ICR mice of 6-8 weeks old were randomly divided into groups A, B and C each with 12. The mice in the 3 groups were subcutaneously immunized with Eg myophilin protein, blank plasmid protein or PBS, respectively, by 3 times and challenged with protoscoleces of E. granulo-sus 2wk after the last vaccination. Mice were sacrificed 20wk after the infection, the hydatid cysts were collected for mea-suring the weight reduction. Spleens were obtained and the splenocytes were separated and cultured in vitro with EgAg or ConA stimulus for 4-5 h. The subsets of CD4+ and CD8+ T cells were measured by FACsort. The proliferation of spleno-cytes was determined by MTT method with blank plasmid and PBS as control.  Results  The average weight of the hy-datid cysts in the immunized group decreased by 69.1% in comparison to the blank plasmid and PBS groups. The CD4+ subset [(29.7±0.9)%] and CD8+ subset [（9.7±0.8）%] in group A increased significantly than group C, [(11.6±1.4)%] and [(7.8±0.2)%] respectively (P<0.01 or <0.05). The ratio of CD4+/CD8+ subsets in group A (3.061±0.015) was also higher than group C (1.487±0.106) (P<0.01). Without stimulation, the proliferation of T lymphocytes in group A（0.237±0.009）was high-er than group C (0.159±0.005) (P<0.01)， with EgAg or ConA stimulus, it was also higher in group A than that of group C(P<0.01).  Conclusion  The recombinant myophilin protein of E. granulosus can induce the proliferation of splenocytes and Th1 response in mice, and the CD4+ T cells subset may bear a part in the induced protection against the challenge of protoscoleces.

Effect of Ketamine on the Development of Chrysomya megacephala （Diptera ∶ Calliphoridae）

LV Zhou，ZHAI Xian-dun，ZHOU Hai-mei，LI Pu，MA Jin-qi，GUAN Ling，MO Yao-nan*

2012, 30(5):  5-361-366. 

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【Abstract】   Objective   To research the pattern of larvae and pupae development when exposed to ketamine.  Methods  The larvae of Chrysomya megacephala were reared in artificial diet containing ketamine with concentration of 0, 25, 50 and 100 mg/kg respectively at 32 ℃, 28 ℃ and 24 ℃ in environmental chamber with a 12-h photoperiod and 75% humidity. 10 samples were collected from each group every 12 h from the 16th h after hatching to pupation. The max body length, weight of the larvae, growth rate of body length, weight and developmental duration of each stage were observed.  Results  The average length and weight in the treatment groups were significantly less than the control before achieving the maximum (P＜0.05）, and the growth rate of 1/2LD50 group at 24 ℃ was most retarded. No dose dependence were observed among the ketamine fed groups. The effect of ketamine dose, temperature and their interaction on larval length and weight was statistically significant (P＜0.01）. The effect of ketamine dose, temperature and their interaction account for, respectively, 20.9%, 60.2% and 18.9% of the total effect on growth of larval length, and they account for 8.3%, 85.6% and 6.1% of the total effect on growth of larval weight. The duration of larval stage in treatment groups was significantly delayed in comparison to the control at different temperatures (P＜0.05）, and the duration of prepupal stage in treatment groups was significantly delayed (P＜0.05）. However, the duration of pupal stage in treatment groups at 24 ℃ was significantly shorter than the control (P＜0.05）.  Conclusion   The time achieving maximum length and weight was significantly delayed, which results in an increased development duration of larval and prepupal stages, indicating that ketamine inhibits the growth of the larvae of C. megacephala.

Study on Hepatocyte Apoptosis of Domestic Pigs Experimentally Infected with Taenia asiatica and Taenia saginata

MOU Rong， BAO Huai-en*， ZHANG Ke，WU Jia-hong， LANG Shu-yuan

2012, 30(5):  6-367-372. 

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【Abstract】   Objective   To investigate apoptosis in liver tissue of the domestic pigs infected with eggs of Taenia asiatica and Taenia saginata.  Methods  The adult worms of T. asiatica and T. saginata were collected and identified from the taeniasis patients in Dunyun and Congjiang districts， Guizhou province. Eggs were collected from gravid proglottids and prepared by washing and centrifugation. Nineteen 20-day hybrid domestic pigs （Duroc-Yorkshire-Landrace strain） were randomly divided into T. asiatica group (6 pigs）， T. saginata group (8 pigs） and control group (5 pigs）. Each animal of experimental groups was infected with 1.5×105 eggs by stomach injection. On day 15， 32， 46 and 74  after infection， animals were sacrificed and liver samples were collected for further experiments. The liver tissues were sliced for glass slides and prepared for ultrathin sections. The apoptosis of hepatocytes was identified by terminal deoxynucleotidyl trans-ferase-mediated dUTP nick and labeling. The morphological features of liver tissue were observed under transmission elec-tron microscope.  Results  The infection rate of two experiment groups reached 100%. Better developed cysticerci were found in liver of T. asiatica group than that of T. saginata group, but the liver pathological changes caused by cysticerci were similar. On day 15 and 32 after infection, hydropic degeneration, obvious vacuolization and some balloon-like degeneration were found in hepatocytes, and focal hepatic necrosis was observed. On day 46, spotty necrosis occurred in some local liver tissues. On day 74, main damages were granulomatous reactions surrounding cysticercus and focal liver fibrosis. On day 46， apoptosis index in T. asiatica group [（15.07±3.42）%] and T. saginata group [(17.13±1.62）%] was considerably higher than that in the control [（9.53±1.06）%] (P<0.05）. On day 74， apoptosis index in T. asiatica group [（27.33±0.92）%] and T. saginata group [(34.20±0.73）%] was higher than that in the control [（13.60±2.26）%] (P<0.05）， and the apoptosis index in T. saginata group was significantly higher than that of T. asiatica group （P<0.05）. Simultaneously， morphological characteristics of apoptosis were clearly observed in hepatocytes in two experimental groups， showing shrunken， wrinkled and deformed nucleus with consolidation of chromosomes and appearance of apoptic body.  Conclusion  The hepatocyte apoptosis of domestic pig is induced in the middle and late stages of infection by the cysticerci of T. asiatica and T. saginata,  indicating that hepatic apoptosis might be related to focal liver fibrosis of the host caused by the cysticerci.

In vitro Killing of Adult Trichinella spiralis by Exogenous Nitric Oxide

WANG Xiao-li1，WANG Yuan-yuan1，FANG Qiang1 *，XUE Yu-qin1，SHEN Ji-long2

2012, 30(5):  7-374-377. 

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【Abstract】   Objective   To investigate the lethal effect of exogenous nitric oxide on adult worms of Trichinella spiralis in vitro.  Methods  Adult worms of T. spiralis isolated from the small intestine of Trichinella-infected BALB/c mice were cultured in RPMI 1640 medium with sodium nitroprusside（SNP） in different final concentration of 0， 0.02， 0.05， 0.10， 0.20， 0.50， and 1.00 mmol/L， 1.00 mmol/L SNP+0.15 mmol/L hemoglobin （Hb）， 1.00 mmol/L SNP+0.15 mmol/L FeSO4， 1.00 mmol/L SNP+1.00 mmol/L L-cysteine （L-cyst）， 1.00 mmol/L SNP+0.15 mmol/L FeSO4 +1.00 mmol/L L-cyst， respectively， and incubated at 37 ℃ in a humidified 5% CO2 atmosphere. On the 4th day after incubation， the adult worms were stained with safranin， and observed under light microscope. The worm mortality in the groups was analyzed.  Results  Under concentration of 0.02 and 0.05 mmol/L， SNP was not cytotoxic to adult T. spiralis with an inhibition of （1.4±1.2）% and （3.2±1.0）%， respectively. The worm mortality in the groups of SNP 0.10， 0.20， 0.50， and 1.00 mmol/L was （9.9±1.8）%， （37.7±2.5）%， （50.1±3.5）%， and （80.8±1.1）%， respectively， significantly higher than that of negative control group ［（1.9±0.2）%， P<0.05］. There was a positive linear  correlation between the worm mortality and SNP concentration in the range of 0.02-1.00 mmol/L. Combination of hemoglobin， L-cyst， FeSO4 and  FeSO4+L-cyst with 1.00 mmol/L SNP led to a decrease of the mortality from （80.8±1.1）% to （56.5±3.7）%， （69.8±2.3）%， （74.8±2.4）%， （72.7±5.6）%， respectively.  Conclusion  Exogenous nitric oxide released from SNP can kill adult worms of Trichinella spiralis. However， hemoglobin and L-cysteine+FeSO4 can reverse its lethal effect on the parasites.

A Cross Analysis on the Theoretical Testing Results on Diagnosis of Common Parasitic Diseases: National Technique Competition for Parasitic Disease Diagnosis in 2011

WANG Qiang1， LI Shi-zhu1， ZHANG Li1， FU Qing1， LIU Wei1， XU Jing1， CHEN Ying-dan1，

2012, 30(5):  8-378-381、386. 

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【Abstract】   Objective   To understand the theoretical level among technicians from disease control and prevention institutions on basic knowledge of parasitic disease diagnosis.  Methods  A national competition on basic knowledge of parasitic disease detection was organized in September， 2011， with 119 technicians participating from disease control and prevention institutions at province， prefecture or county level in 30 provinces. Database was constructed with the essential information of participants and scores of testing results. T-test or one-way Anova methods were used to analyze the scores by gender， age， professional title， insititutions and places of participants.  Results  The scores of the competitors were in the range of 40-93 with an average value of 66.37±11.80 and a passing rate of 74.95%. No difference was shown by gender， age and professional title（P>0.05）. Except the knowledge of helminth detection, the score on the knowledge in other aspects was higher among the technicians from provinces with control activities of schistosomiasis and other parasitic infections than those from provinces without the control activities (P<0.05). The same was true in technicians from areas with or without malaria control activities (P<0.05).  Conclusion  The level of basic knowledge on parasite detection is low in general among technicians at the institutions of parasitic disease control, and therefore, training needs to be strengthened.

Influence Factors of Enterobius vermicularis Infection among Pupils in Chongqing City

WU Cheng-guo1 *，XIE Jun1，LUO Xing-jian1，LEI Qun-jian2，LIU Qing-rong1，XIAO Bang-zhong1，LI Shan-shan1

2012, 30(5):  9-382-386. 

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【Abstract】   Objective   To investigate Enterobius vermicularis infection among primary school students and its influence factors in Chongqing. Methods  Beibei and Changshou were selected as investigated points from October to December 2011. One primary school was randomly chosen from each of the 5 different directions in every investigated point. Adhesive cellophane anal swab was used to examine pinworm eggs for 3 consecutive days. Information on children′s family， hygiene habits and school environment was collected by questionnairing．  Results  The total infection rate of E. vermicularis was 6.7%(71/1 071）. The infection rate in rural schools (7.9%， 60/755） was higher than that of urban schools (3.8%， 12/316） (χ2=6.1169， P<0.05）. The rate in males and females was 6.3%(34/536） and 7.1%(38/535）， respectively (χ2=0.2463， P>0.05）. Among the investigated children aged 6~12 years, the infection rate in 6-year-old children (16.03%） was highest. There was a statistical significance among age groups (χ2=29.1492， P<0.01）. With the increase of age， the rate decreased. Multivariate logistic regression analysis showed that location (OR=0.411）， age groups (OR=0.714）， education level of mothers (OR=0.568）， materials of classroom-ground (OR=0.116） and types of boarding (OR=0.272） were the influence factors on E. vermicularis infection in primary schools (P<0.05）.  Conclusions  Pinworm control should more focused on rural children， younger group， mothers with lower education， classroom with cement ground and lodging schools in Chongqing City.

Epidemiological Analysis of Imported Malaria in Henan Province in 2011

CHEN Wei-qi*，SU Yun-pu，DENG Yan，ZHANG Hong-wei

2012, 30(5):  10-387-390. 

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【Abstract】   Objective   To analyze the epidemiological characteristics of imported malaria in Henan Province in 2011， and provide scientific basis for the control of imported malaria.  Methods   Data of imported malaria were collected and analyzed by SPSS11.5 in 2011.  Results  145 imported malaria cases were reported. Among them， 110 （75.9%） were falciparum malaria， 31 （21.4%） were vivax malaria， 3 （2.1%） were quartan malaria， and 1 （0.7%） was ovale malaria. The ratio of males to females was 47.3 ∶ 1 （142/3）. The average age was （37.7±9.4） years old. 113 patients returned from Africa， accounted for 77.9% of the total. The main reason for going abroad among the cases was labor export （82.8%， 120/145）. The cases were reported every month. The median interval from symptom appearing to diagnosis was 4 d， only 19 patients （13.1%） were diagnosed within 24 h. Eighty-six cases （63.2%） were reported by municipal medical units and CDC. Forty cases （29.4%） were reported by county level medical units and CDC. A total of 143 cases recovered with chemotherapy and two cases died.  Conclusion  In order to avoid the death of malaria cases and reduce the risk of secondary transmission， malaria screening and health education for those returned from malaria-endemic areas should be strengthened and the diagnosis and treatment capabilities be improved at county medical units.

Malaria Conjugate Vaccine

QIAN Feng，XU Hu-ji

2012, 30(5):  11-393-395、400. 

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【Abstract】  Many malarial antigens are poor immunogens in human. Chemical conjugation is one of the ways to enhance the immunogenicity of those poor immunogens. Its value has been demonstrated in malaria vaccine research. This article introduces the chemical linkers and the carrier proteins, which are often used for protein conjugation, and summarizes the research progress in malarial conjugate vaccine development.

Immune Evasion Molecules of Toxoplasma gondii

ZHENG Bin，LU Shao-hong*

2012, 30(5):  12-396-400. 

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   【Abstract】  Toxoplasma gondii can live in the host for a long time relying on effective immune escape mechanisms and cause a chronic infection. Different antigenic molecules in the parasitophorous vacuole membrane， rhoptry and cytoplasm play an important role in evading the host′s immune response. They can effectively help avoid the host immune response by inhibiting excessive inflammatory response and production of nitric oxide， affecting gene expression of some cytokines， depleting the specific immune substances， reducing the function of immune cells and so on. This paper summarizes the latest research advances in molecules related to the immune evasion of T. gondii.

Research Advances in Interplay of Host Immune Mechanism and Echinococcus multilocularis Metacestodes

LOU Zhong-zi，LI Hong-min，YAN Hong-bin，NI Xing-wei，JIA Wan-zhong*

2012, 30(5):  13-401-405. 

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【Abstract】  Through affecting on host innate and acquired immune responses, Echinococcus multilocularis orchestrates various interplays that are beneficial not only to facilitate its intrahepatic proliferation and maturation during life cycle, but also to limit pathological process in its intermediate host. This review reveals the role of the metacestode′s immune-related molecules in modulating host responses and optimizing its own survival.

Research Progress on the Diagnosis Technology of Canine Visceral Leishmaniasis

DING Dan， WANG Jun-yun*

2012, 30(5):  14-406-410. 

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 【Abstract】  Canine visceral leishmaniasis is caused by Leishmania infantum. Infected dogs， either symptomatic or asymptomatic, are considered as the major reservoirs for zoonotic visceral leishmaniasis. Accurate and rapid detection of canine leishmanial infection is crucial for control of human visceral leishmaniasis due to its role in the transmission of the infection to vectors. Various techniques based on parasitology， immunology and molecular biology have been studied and evaluated for detecting canine leishmanial infection. This article reviews the progress in techniques and methods for its diagnosis.

How to Establish the Monitoring and Evaluation System in Elimination Stage of Malaria in China：The Experience from Malaria Indicator Survey System

DUAN Meng-juan 1，DUAN Ji-hui 2，HE Guo-ping 1 *

2012, 30(5):  15-411-414. 

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 【Abstract】   Malaria is one of the most important public health problems around the world. Most region of China has embarked a transition from interruption of malaria transmission to elimination. This paper summarizes the main function and construction, the key parts, and the advantages in national field work of Malaria Indicator Survey (MIS） system, indicates the difficulties of establishing a Monitoring and Evaluation (M&E） System in malaria elimination in China and draws lessions from the MIS.

Application of Flow Cytometry in the Detection of Mouse Malaria Parasites in Living Cells

LIU Yi， XI Wei

2012, 30(5):  16-340-342. 

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Peripheral blood samples were obtained from Plasmodium berghei-infected mice， and stained with Vybrant DyeCycle Green and anti-mouse CD71 PE. With normal mouse as negative control， the blood samples were tested by flow cytometry at 0， 30， and 60 min after staining. There was no positive signal in the erythrocytes from negative control and the unstained erythrocytes from the infected mouse. However， the positive signal was detected in the stained erythrocytes from the infected mouse. The results showed an adequate consistency by intra-class correlation coefficient （ICC） test （ICC=0.999， P<0.01）. Flow cytometry with the use of Vybrant DyeCycle Green and anti-mouse CD71 PE in staining living cells can be used for the detection of malaria parasite-infected blood. In a given period， there is no negative impact on the detection of unfixed cells.

Observation on Salivary Gland Polytene Chromosomes of Simulium qianense （Diptera ∶ Simuliidae）

HUANG Li1，ZHANG Chun-lin1 *，HUANG Ruo-yang2，CHEN Han-bin1

2012, 30(5):  17-353-356. 

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 【Abstract】  The salivary gland polytene chromosomes of Simulium qianense was observed with wild larvae collected from Guiyang， Guizhou Province. Using an upgraded method of preparation， the polytene chromosomes of the larval salivary glands of S. qianense were observed and analysed. The results showed three pairs of chromosomes （2n=6） arranged from the longest to the shortest. Chromosome Ⅰ was metacentric while chromosomes Ⅱ and Ⅲ were submetacentric. The centromeric regions distinctly expanded and were easily recognized. The nucleolar organizer situated near the centromeric band of the short arm of chromosome Ⅱ. The Balbiani ring and double bubble all located near the tip of the short arm of chromosome Ⅱ. There were inversions in some individuals, with an inversion frequency of 33% （32/97）. The study revealed constant localization and morphology of the principal landmarks （namey， centromere， nucleolar organizer， Balbiani ring， double bulge， band and puff） which can be considered as distinguishable features of S. qianense.

Scanning Electron Microscope Observation of Plerocercoid from  Rana nigromaculata in Guizhou Province

XU Jing1，CHEN Yan1 *，TANG Gui-wen1，LIU Xian-lin2

2012, 30(5):  18-372-373. 

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【Abstract】  Five integral and active plerocercoids collected from naturally infected frogs were fixed by glutaraldehyde?-osmic acid， dehydrated in graded series of ethanol， dried via vacuum freeze and coated with carbon gold. They were then observed by scanning electron microscopy. The results showed that the anterior and posterior ends of the plerocercoids were slightly swell and without segmentation. Annular furrows ran irregularly along the body surface. The anterior end was drawn inside and had a deep dorsoventral hollow， around which， the plasma membrane displayed a lip bulge. The posterior end was similar to the anterior one in the shape， but a fissure in its central part was comparatively narrower and shallower than the hollow in the anterior end. There were thousands of pits and grooves on the body surface, unequal in size. The whole body surface of the plerocercoid was densely covered by sharp spine-like microtriches.

Investigation on Babesia in Ticks Infested on Police Dogs in Selected Areas of China

WEI Fu-rong1， LAN Qin-xian1， ZHU Dan1，YE Jun-hua2， LIU Qin1， ZHANG Yi1 *

2012, 30(5):  19-390-392. 

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【Abstract】  399 tick specimens were collected from the body surface of police dogs in Chongqing municipality，  provinces of Fujian， Guangdong， Hainan， Guangxi， Hebei， Henan， Shanxi， Jiangsu and Zhejiang. Nested-PCR and se-quence testing were taken to investigate the prevalence of Babesia sp. in ticks.  The results showed that Babesia vogeli was found in ticks infested on the body surface of police dogs， with a positive rate of 5.3%. The prevalence in Chongqing， Guangdong， Guangxi， Hainan and Zhejiang was 4/16， 3.6%（1/28）， 12.5%（11/88）， 3.3%（4/121） and 1/15， re-spectively. It suggested that there was a certain rate of infected ticks infested on the body suriface of police dogs, which contrubuted to the potential threat to staff. The prevention and control measures should be strengthened.

Preparation of Agar-Paraffin Double-Embedded Longitudinal Sections of Schistosoma japonicum

WANG Ke-geng1，ZENG Qing-ren2，ZHANG Yu-kuai1，ZHOU Jun3，CAI Li-ting，LIANG Yu1，LIU Yan1,2 *

2012, 30(5):  20-415-417. 

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【Abstract】  Schistosoma japonicum adults are pre-embedded in a double-layer agar and made the block, then dehydrated with alcohol, isobutyl alcohol and n-butyl alcohol. Various staining procedures can be conducted after conventional sectioning and dewaxing. Complete longitudinal serial sections of the pre-embedded worms can be obtained, and the desired sections can be easily located accurately.

Serological Investigation of Toxoplasma gondii Infection in HIV Positive Cases in Dali and Dehong of Yunnan

YOU Ying-xia1，LI Wei1，SHEN Li-jie1，2 *，NIE Da-ping1

2012, 30(5):  21-418-419. 

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 【Abstract】  Serum samples were collected from HIV positive cases （927） and HIV negative ones （80） from June 2010 to August 2011 in Dali and Dehong Prefectures of Yunnan. Serum anti-Toxoplasma gondii IgG was detected by ELISA. The overall anti-Toxoplasma gondii IgG positive rate among HIV positive cases  and HIV negative ones was 35.1% （325/927） and 23.8％ （19/80）， respectively. In HIV positive cases, the seropositive rate was 30.3％ （178/588） in Dali and 43.4％ （147/339） in Dehong. The seropositive rate was significantly different among ethnic groups (χ2=28.433， P<0.05）. No significant difference was found among age groups(χ2=4.248，P＞0.05）, and the age group of 41-60 showed the highest positive rate (36.1%， 103/285）. The seropositive rate was 35.6%（203/571） in males and 34.3% (122/356） in females (χ2=0.158，P＞0.05）.