[Effect of comprehensive care for chronic filariasis patients with lymphedema].

OBJECTIVE: To evaluate the effect of chronic filariasis patients with lymphedema after comprehensive cared. METHODS: A total of 386 chronic filariasis patients with lymphedema received the comprehensive care including soaking feet by TCM, washing feet by single Chinese medicine or clear water, preventing and eliminating invasive wound, physical training, raising the limb, and wearing suitable shoes. The attack frequency of inflammation of lymphatic vessels, the stage of lymphedema disease, and leg circumference were observed before and after the care. RESULTS: After the comprehensive care, the attack rates of inflammation of lymphatic vessels decreased from 9.65% to 4.67%, and there was a significant difference (χ2 =7.34, P=0.0067). The proportion of the patients with high stage of lymphedema disease decreased significantly (all P <0.01), and the average leg circumference decreased of 1.06 cm. The skin appearance improved significantly. Other signs, such as pitting edema, ulcer, and skin folds also improved significantly (all P <0.01). CONCLUSION: The comprehensive care for chronic filariasis patients with lymphedema has a remarkable effect and is worthy of further application.

Pontibacter yuliensis sp. nov., isolated from soil.

A Gram-staining-negative, rod-shaped and pink bacterium was isolated from the soil of a Populus euphratica forest in the Taklamakan desert, Xinjiang, China. It was designated strain H9X(T). A 16S rRNA gene sequence homology search indicated that the isolate was most closely related to the family Cytophagaceae. The 16S rRNA gene of strain H9X(T) displayed 94.2-96.3 % sequence identities to those of type strains of other species of the genus Pontibacter. It only possessed menaquinone-7. The major cellular fatty acids of the novel isolate were iso-C15 : 0, C16 : 1ω5c summed feature 3 (containing C16 : 1ω6c and/or C16 : 1ω7c) and summed feature 4 (comprising anteiso-C17 : 1 B and/or iso-C17 : 1 I). The major polar lipids were phosphatidylethanolamine, one unknown aminophospholipid, one unknown glycophospholipid and several unknown phospholipids. The DNA G+C content of this bacterium was 55.2 mol%. Based on the phenotypic and genotypic data presented, it can be concluded that this isolate represents a novel species of the genus Pontibacter, for which the name Pontibacter yuliensis sp. nov. is proposed. The type strain is H9X(T) ( = CCTCC AB 2013047(T) = KCTC 32396(T)).

Genomic and functional characterization of three new venom peptides from the scorpion Heterometrus spinifer.

Three new cysteine-free venom peptides, which are referred to as Heterin-1, Heterin-2 and Spiniferin, respectively, were identified from the scorpion Heterometrus spinifer. Heterin-1, Heterin-2 and Spiniferin contain 43, 24 and 13 amino acid residues, respectively. Genomic analysis showed that the genomic organizations of the three peptides are consistent with those of the known Na(+), K(+) or Cl(-)-channel specific toxins from scorpions; this suggests that the genes of the cysteine-free and cysteine-rich peptides from scorpions were derived from a common ancestor. Antimicrobial assay demonstrated that Heterin-1 possesses potent activities against both Gram-positive and Gram-negative bacteria. Among the tested bacterial species, Heterin-1 is the most active against Bacillus megaterium and Micrococcus luteus with MICs of 4.0 µM and 4.0 µM, respectively. Heterin-2 is able to potently inhibit the growth of Gram-positive bacteria with MICs from 5.6 µM to 30.0 µM; however, it has weaker activities against the tested Gram-negative bacteria. It is interesting to see that deletion of the C-terminal random coiled tail (KKD) in Heterin-2 markedly changed the antimicrobial specificity and activity of the peptide. Spiniferin has very weak antimicrobial activities against both Gram-positive and Gram-negative bacteria. We found that introducing three net charges into the polar face of Spiniferin significantly increased its antimicrobial activity against the majority of the tested bacteria; however, in some instances, net charge on the polar face is not important for the antimicrobial activity of the peptide. These studies have expanded our understanding of the diversity, evolution and structure/function relationships of the cysteine-free peptides from scorpions.

[Prediction of hookworm incidence with time-series model in Jiangsu Province].

OBJECTIVE: To explore the feasibility of autoregressive integrated moving average (ARIMA) to predict the infection rates of hookworm in Jiangsu Province. METHODS: From 1990 to 2006, the infection rates of hookworm were used for a training data set. As to obtain a stationary data set, the training data set was second-order differenced using the version SAS 9.0. The model parameters were screened by using the minimum information criterion. The ARIMA model was constructed to predict the infect rates of hookworm form 2007 to 2011. RESULTS: The time-series model ARIMA (1, 2, 0) was confirmed preliminarily. The model fitted well the training data set. The predictive infection rates were main accordance with the actual status of hookworm from 2007 to 2011, and the most minimum error was only 9.23%. CONCLUSION: The model constructed has a good predictive effect and applied value for control of hookworm.

Three new antimicrobial peptides from the scorpion Pandinus imperator.

Three novel cysteine-free venom peptides, which were referred to as Pantinin-1, Pantinin-2 and Pantinin-3, respectively, have been identified from the scorpion Pandinus imperator by cDNA cloning strategy. The precursor of each peptide consists of a signal peptide, a mature peptide with no disulfide bridges, and an acidic propeptide with a typical processing signal. Each of the three peptides is an α-helical, cationic and amphipathic molecule with 13 or 14 amino acid residues. Their amino acid sequences are homologous to those of some 13-mer antimicrobial peptides isolated from scorpions. Antimicrobial assay showed that all the three peptides possess relatively strong activities against Gram-positive bacteria and a fungus, but have very weak antimicrobial activities against Gram-negative bacteria. Toxicity assay showed that the three peptides exhibit very low or mild hemolytic activities against human red blood cells. It is interesting to see that Pantinin-3 is able to potently inhibit the growth of vancomycin-resistant Enterococcus (VRE) S13, a pathogen that can cause a number of human infections; this suggests that Pantinin-3 has great potential to be applied in the treatment of VRE infections. Our findings gain new insights into the structure/function relationships of the small linear cationic antimicrobial peptides from scorpions, and provide new templates for designing of antimicrobial agents targeting antibiotic-resistant pathogenic bacteria.

Molecular and bioinformatical characterization of a novel superfamily of cysteine-rich peptides from arthropods.

The full-length cDNA sequences of two novel cysteine-rich peptides (referred to as HsVx1 and MmKTx1) were obtained from scorpions. The two peptides represent a novel class of cysteine-rich peptides with a unique cysteine pattern. The genomic sequence of HsVx1 is composed of three exons interrupted by two introns that are localized in the mature peptide encoding region and inserted in phase 1 and phase 2, respectively. Such a genomic organization markedly differs from those of other peptides from scorpions described previously. Genome-wide search for the orthologs of HsVx1 identified 59 novel cysteine-rich peptides from arthropods. These peptides share a consistent cysteine pattern with HsVx1. Genomic comparison revealed extensive intron length differences and intronic number and position polymorphisms among the genes of these peptides. Further analysis identified 30 cases of intron sliding, 1 case of intron gain and 22 cases of intron loss occurred with the genes of the HsVx1 and HsVx1-like peptides. It is interesting to see that three HsVx1-like peptides XP_001658928, XP_001658929 and XP_001658930 were derived from a single gene (XP gene): the former two were generated from alternative splicing; the third one was encoded by a DNA region in the reverse complementary strand of the third intron of the XP gene. These findings strongly suggest that the genes of these cysteine-rich peptides were evolved by intron sliding, intron gain/loss, gene recombination and alternative splicing events in response to selective forces without changing their cysteine pattern. The evolution of these genes is dominated by intron sliding and intron loss.

[Comparison between two expression ways on cysteine protease of Clonorchis sinensis].

OBJECTIVE: To compare the prokaryotic expression system and eukaryotic expression system for the expression of cysteine protease of Clonorchis sinensis, and the diagnostic efficiency of their objective products. METHODS: According to the sequence of cysteine protease of C. sinensis, two pairs of primers were designed to amplify the genes from the total cDNA of C. sinensis. The genes were cloned into plasmid pET28a (+) and pPIC9K, respectively, and these recombinant plasmids were transformed into E. coli BL21 and GS115 separately after they were identified through double digests and sequencing. The cysteine protease of C. sinensis was expressed and purified, and then the sero-diagnostic effects of the purified proteins for clonorchiasis by ELISA were compared. RESULTS: The cysteine protease of C. sinensis was expressed as inclusion bodies in BL21, and its yield was 6.8 mg/L, while it was expressed as a kind of soluble protein in GS115, and its yield reached to 65.00 mg/L. Their sensitivities for serodiagnosis of clonorchiasis were 95.00% and 93.30%, respectively, and their specifities were 91.67% and 94.10%, respectively, with no statistically significant differences between them (all P values were above 0.05). CONCLUSION: The application value on cysteine protease of C. sinensis expressed through eukaryotic expression system is higher than that expressed through prokaryotic expression system.

Tremendous intron length differences of the BmKBT and a novel BmKBT-like peptide genes provide a mechanical basis for the rapid or constitutive expression of the peptides.

The cDNA sequence encoding a novel BmKBT-like peptide (referred to as BmKBy) was cloned and sequenced from the scorpion Mesobuthus martensii Karsch. Functional analysis indicated that both BmKBT and BmKBy possess strong toxicity in mice, but very weak toxicity in cotton bollworm. Phylogenetic analysis showed that BmKBy and BmKBT represent evolutionary intermediates between the α- and ß-toxins from scorpions. The genomic sequences of BmKBT and BmKBy were also obtained. It is interesting to see that two genes, which contain an intron of 225 and 1529bp, respectively, exactly code for the BmKBT peptide. One gene, which contains an intron of 1312bp, codes for BmKBy. Given that genes with long introns favor constitutive expression, whereas those with short introns are rapidly regulated in response to stimulations, the BmKBT_a and BmKBT_b genes provide a mechanical basis for either constitutive expression or rapid generation of the toxic peptides in response to different signals.

[Epidemiological survey of echinococcosis in some areas of Jiangsu Province].

OBJECTIVE: To understand the prevalence of echinococcosis in some areas of Jiangsu Province, so as to provide the evidence for making practical control measures. METHODS: The cases were determined by a retrospective survey and case-clues survey. All canine feces were tested by immunological fecal antigen in the villages of the local infection suspected cases. The antigen-positive feces were detected for E. chinococcus eggs by Kato-Katz technique. A serological investigation was administrated to search infected persons by double methods of enzyme-labeled immune assay and point immuno-gold filtration assay in the focus groups around the local infection suspected cases, and some students aged from seven to twelve years in their townships and in five different-orientation townships sampled randomly in their counties. The antibody-positive people were confirmed further by using imaging detection. RESULTS: There were 10 imported cases and 16 local infection suspected cases. Twenty canine feces were positive among 1 938 samples, and the positive rate was 1.03%. However, no pathogen was found. Of 12 473 serum samples, 72 were positive in sera immunological antibody test (0.58%), but no cystic nodules were found by imaging detection. CONCLUSION: There is no direct evidence for confirming the presence of infectious source and foci of echinococcosis in Jiangsu Province. However, the surveillance of echinococcosis is still needed.

[Surveillance of soil-transmitted nematodiasis in northern Jiangsu Province from 2006 to 2010].

OBJECTIVE: To evaluate the control effect on soil-transmitted nematodiasis in surveillance sites of northern Jiangsu Province. METHODS: According to "The Monitoring Program on Soil-transmitted Nematodiasis in Jiangsu Province", the eggs of soil-transmitted nematodes were detected by the Kato-Katz technique and cellophane anal swab technique, and the infection rates were predicted by the horizontal average speed development method in the surveillance sites from 2006 to 2010. RESULTS: The overall infection rate of soil-transmitted nematodes declined to 3.13% and the intensities of the infections were mild in past 5 years in the northern Jiangsu Province, and the infection rates of Ascaris lumbricoides, hookworm, Trichuris trichiura and Enterobius vermicularis were 0.77%, 0.89%, 1.49 and 1.19%, respectively. The overall infection rates showed a downtrend. The predicted result also demonstrated that the future infection rates should keep a continuous downtrend, but the decreasing speed should be slower than before. CONCLUSION: The infections of soil-transmitted nematodes in the northern Jiangsu Province have been controlled basically, and we should adjust the control strategy including surveillance and chemotherapy in the main risk population.

[Epidemic situation and control of soil-transmitted nematodiasis in Jiangsu Province].

OBJECTIVE: To analyze the prevalence changes and the control measures of soil-transmitted nematodiases in Jiangsu Province in the recent 20 years and evaluate the control effect, so as to explore the control strategies and measures appropriate to current epidemic characteristics. METHODS: The data on surveillance, control measures and control effect of soil-transmitted nematodiases in Jiangsu Province from 1990 to 2009 were collected and analyzed. RESULTS: Different control programs were taken in different control stages in Jiangsu Province. A total of 106 916.6 thousand person-times accepted chemotherapy and 2 042.9 thousand person-times were surveyed in the last 20 years. The infection rate of soil-transmitted nematodiases was 1.31% in 2009, which decreased by 97.79% comparing with 59.32% in 1990. The results showed that the control effectiveness was significant. CONCLUSIONS: The epidemic situation of soil-transmitted nematodiases has been controlled effectively in Jiangsu province. In order to consolidate the control achievements, the control strategies should put emphasis on health education, water supply and sanitation and environment sanitation improvement, meanwhile, the surveillance and medication in key population should continue to be strengthened, as well as soil-transmitted control among migrant population.

[Process for lymphatic filariasis elimination in Jiangsu Province, China].

OBJECTIVE: To review the process of lymphatic filariasis elimination in Jiangsu Province, and summarize the control experience. METHODS: The data of epidemic status, control and surveillance of lymphatic filariasis were collected and analyzed in Jiangsu Province from 1950s to 1990s. RESULTS: Lymphatic filariasis was endemic in 71 counties (cities) among all 75 counties (cities) in Jiangsu Province. Filariasis bancrofti distributed widely in the province, and in some areas of the south of Jiangsu, there were filariasis bancrofti and Brug' s filariasis co-existing. Before 1971, the microfilaria rate (mfr) in some areas had ever reached 20. 15% , from 1979, large scale control programme was carried out and the mfr in all endemic villages decreased to below 1% , and in 1989, the rate dropped to 0.016%. By 2001, the whole regions of the province reached the criteria of filariasis elimination. CONCLUSION: The elimination of lymphatic filariasis in Jiangsu Province provides valuable experience for lymphatic filariasis control in other areas.

[Performance evaluation of rural latrine renovation in key schistosomiasis control areas, Jiangsu Province].

OBJECTIVE: To evaluate the hygienic, environmental, social and economic benefits achieved by rural latrine renovation in key schistosomiasis control areas, Jiangsu Province. METHODS: Eights villages (evaluation villages) that finished and the three villages (control villages) that did not finish the latrine renovation work were sampled and investigated through field visits, questionnaire, referring to information and so on, to collect relevant data, and human waste samples were detected for laboratory indicators in hygiene and environment, in key schistosomiasis control areas in three counties (cities, districts), Jiangsu Province. RESULTS: A total of 11 villages and 220 households were investigated. In the evaluation villages, the owning rate and quality conformity rate of three-format sanitary latrines were 98.6% and 98.8%, respectively. The human schistosome infection rate, intestinal parasite infection rate, the incidence of intestinal infectious diseases, and related medical costs decreased by 100%, 44.5%, 34.2% and 82.8%, respectively, compared with those before latrine renovation. In laboratory testing, the removal rates of fecal coliform values, CODcr, BOD5 and ammonia nitrogen in the third cell of household latrine were 99.99%, 68.50%, 63.17% and 52.30%, respectively, compared to those in the first cell. The village appearance had changed fundamentally, the villagers were satisfied with the latrine renovation, and their health knowledge and health behavior improved significantly. CONCLUSIONS: Rural latrine renovation in key schistosomiasis control areas has got great achievements in hygienic, environmental, social and economic benefits. It plays an important role in the promotion of schistosomiasis control, economic development and social harmony.

[Cloning, expression and evaluation on effect in serological diagnosis of cysteine protease of Clonorchis sinensis].

OBJECTIVE: To clone and express the cysteine protease of Clonorchis sinensis and evaluate its effect on immunodiagnosis of human clonorchiasis. METHODS: Based on a cysteine protease gene fragment of C. sinensis (CS-CP, GenBank accession: AF093242), a pair of primers were designed and amplified from total cDNA of C. sinensis, and the gene was cloned into plasmid pPIC9K and expressed in Pichia pastoris GS115. The expressed product was purified. Ten BALA/c mice were immunized with the purified CS-CP, and the anti CS-CP antibody in the sera of immunized mice was tested with ELISA. Finally, its effect on serodiagnosis was evaluated with Dot-ELISA and Western blot. RESULTS: The CS-CP gene (approximately 927 bp) was successfully amplified, cloned and expressed in Pichia pastoris GS115. The sera of mice immunized with purified rCS-CP could be recognized by the soluble antigen of C. sinensis adult worms and its special anti CS-CP antibody titer was high (1: 64 000). The rCS-CP was probed by Dot-ELISA and Western-blot with sera from patients with clonorchiasis and other parasitic infections, and it had a sensitivity of 91.7% (55/60) in diagnosis of clonorchiasis and a specifity of 97.6% (82/84) for healthy population. The rCS-CP had no cross-reaction with the patients of schistosomiasis japonica, but had a cross-reactivity of 20.0% (2/10) with the patients with paragonimiasis westermani. CONCLUSIONS: rCS-CP possesses a favorable diagnostic effect and it is a better serodiagnostic one among recombinant antigens.