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28 February 2023, Volume 41 Issue 1

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SPECIAL REPORT

Forty years of the Chinese Journal of Parasitology and Parasitic Diseases since its publication

SHENG Huifeng, ZHOU Xiaonong, YU Senhai, TANG Linhua, FENG Zheng, LI Shizhu, XUE Chunliang, WU Guanlin, YU Xinbing, WEN Tinghuai, CHENG Xunjia, PAN Weiqing, HU Wei, SU Chuan, WANG Tianping, WU Zhongdao, CHEN Qin, ZHANG Zhengyan, DAI Jing, LI Di, LIU Yuzhou, CAO Jianping

2023, 41(1):  1-9.  doi:10.12140/j.issn.1000-7423.2023.01.001

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The Chinese Journal of Parasitology and Parasitic Diseases had its first issue in 1983. Over the past 40 years, the journal witnesses the remarkable achievements in prevention and control of important parasitic diseases in China, marked by effective control of schistosomiasis (2015) and elimination of lymphatic filariasis (2007) and malaria (2021), and has been playing an important role providing academic exchange platform for research and control programs of parasitic diseases and parasitology, promoting domestic and international scientific exchanges. With the guidance of each editorial board, the journal strictly abides by the academic standards and perseveres the concept of presenting high-quality publication. The national and international impact of the journal has continuously increased. It has been included in important international databases such as Medline, Scopus, and Chinese Science Citation Database (CSCD) (Class C) since 1989. It has been selected as one of Chinese Science and Technology Core Journals since 1987, Core Journals in Chinese since 1992 and the World Journal Clout Index Report Scientific & Technological Periodicals (WJCI) since 2020, and was included in the High-Quality Scientific and Technological Journal Classification Catalog for Preventive Medicine and Health (2021 edition). It was also awarded as one of “the 100 Outstanding Chinese Academic Journals” (in 2009), “the First Prize of Excellent Medical and Health Journals of the Ministry of Health” (in 2005), “the First Prize of Excellent Journals of the National Health and Family Planning Commission” (in 2013), the First Prize and the Excellent Journal of “the Journal series of the Chinese Preventive Medicine Association” and “the Excellent Journal Award in East China Region”. In 2021, its core impact factor reached 1.743, the highest in its history, and continues its first rank among 31 core journals in medicine. The network informatization has been constructed since 2006, with "WeChat" platform established in 2016, which has raised the journal’s dissemination, service capacity and impact to a higher level. This article summarizes the 40-year history of the Chinese Journal of Parasitology and Parasitic Diseases, and prospects its future development direction.

EXPERT VIEWPOINT

Research progress on the effect of human volatiles on mosquito behavior

ZHANG Hong, CHENG Gong

2023, 41(1):  10-14.  doi:10.12140/j.issn.1000-7423.2023.01.002

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Mosquitoes are vectors of mosquito-borne infectious diseases such as malaria, dengue, Zika, and yellow fever. Mosquitoes detect the presence of hosts by integrating chemosensory, thermal, and visual cues. Among these, volatiles is crucial for host detection in mosquitoes. In this review, we summarized the sources and factors of human volatiles and their effects on mosquito behavior, which may provide a new approach to the development of mosquito attractants and repellents.

ORIGINAL ARTICLES

Anti-Echinococcus multilocularis effect of total alkaloids of Sophora moorcroftiana in water solution and tablet forms in vitro and in vivo

DU Tao, HU Chunhui, GAN Xuehui, GAO Pan, ZHANG Fabin

2023, 41(1):  15-22.  doi:10.12140/j.issn.1000-7423.2023.01.003

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Objective To evaluate the anti-Echinococcus multilocularis effect of total alkaloids of Sophora moorcroftiana (TA-SM) in water solution and tablet forms in vitro and in vivo. Methods The protoscolices of E. multilocularis were isolated from the Mongolian gerbils and cultured in a culture flask (10 000 protoscolices/flask). The culture flasks were randomly assigned into the culture medium control group, solvent control group, the albendazole (ABZ) positive control group, the albendazole sulfoxide (ABZ-SO) positive control group, and the low, medium and high concentration groups of TA-SM as water solution groups and as tablet form groups, in which the protoscolices were co-cultured with RPMI 1640 complete medium, or the final concentration of 0.1% dimethyl sulfoxide (DMSO), 10 μg/ml ABZ, 10.60 μg/ml ABZ-SO, or the final concentration of 5 μg/ml (low concentration), 10 μg/ml (medium concentration) and 20 μg/ml (high concentration) of TA-SM in water solution and tablet forms at 37 ℃ and 5% CO₂ for 7 days. The samples collected every day were stained with trypan blue and observed under inverted microscope to check the survival and calculate the survival rate. The relative expression level of alkaline phosphatase (ALP) in the culture supernatant and caspase 3 in protoscolices was detected on the fifth day of co-culture. Fifty-four female Kunming mice were injected with protoscolices of E. multilocularis (0.2 ml/mouse, 10⁴ protoscolices/ml) intraperitoneally and were randomly assigned into the normal saline group, tablet group and water solution group 4 months post-infection. In the water solution group and tablet group, mice were treated with 28 mg/(kg·d) of TA-SM in water solution or tablet forms, respectively, by oral gavage, once a day. In the normal saline group mice were given the same volume of normal saline only. On the 15th, 30th and 60th days of treatment, six mice in each group were randomly selected to dissect, the mass of cyst was weighed, and the rate of cyst inhibition was calculated. The liver tissue sections were prepared, and the pathological changes of the liver tissue were observed using HE staining, the ultrastructure of liver tissue was observed by scanning electron microscope (SEM). One-way ANOVA was used for inter-group comparison, and LSD-t test was used for pairwise comparison. Results On the 7th day of co-culture in vitro, the protoscolices in the medium and in the solvent control groups were oval in shape, and the protoscolices were intact and clear without damage, showing typical invagination. In the ABZ and ABZ-SO and positive control group, the protoscolices became longer, and the sucker was visible, showing the typical eversion type. In the low and medium concentration groups of TA-SM in water solution and tablet forms, the protoscolices were mainly invaginated, and occasionally everted. The morphology of the protoscolices in the high concentration groups of TA-SM in water solution and tablet forms was mostly eversion type. On the 7th day of co-culture in vitro, the survival rates of the protoscolices in the low, medium and high concentration groups of TA-SM in water solution and tablet forms were (34.7 ± 2.75)%, (31.5 ± 10.12)%, (27.6 ± 2.51)% and (30.8 ± 4.15)%, (29.8 ± 8.11)%, (25.2 ± 4.30)%, respectively, with no statistically significant difference between the TA-SM in water solution group and tablet form group (t = 1.36, 0.22, 0.83, all P > 0.05). The survival rates in the TA-SM in water solution group and tablet form group were all lower than that of the medium control group [(78.4 ± 9.68)%] (F = 32.30, 38.53; all P < 0.01) and the solvent control group [(82.7 ± 5.45)%] (F = 55.15, 67.50; all P < 0.01), but there were no significant differences with ABZ positive control group (23.85 ± 19.63)%] (F = 0.53, 0.29; all P > 0.05) or the ABZ-SO positive control group [(18.93 ± 3.08)%] (F =4.44, 3.17; all P > 0.05). On the fifth day of co-culture in vitro, the relative expression levels of ALP in the culture supernatant in the low, medium and high concentration groups of the TA-SM in water solution groups and the tablet form groups, the ABZ and ABZ-SO positive control groups, the medium and solvent control groups were 0.15 ± 0.01, 0.17 ± 0.03, 0.18 ± 0.03 and 0.21 ± 0.04, 0.29 ± 0.04, 0.32 ± 0.04; 0.17 ± 0.01 and 0.15 ± 0.01; 0.15 ± 0.01 and 0.13 ± 0.01, respectively, in which the TA-SM in tablet form groups were higher than medium and solvent control groups, ABZ and ABZ-SO positive control groups (F = 18.49, 22.34, 14.86, 18.06; P < 0.01) and the TA-SM in water solution groups (t = 6.86, 3.78, 5.19; P < 0.05). On the fifth day of co-culture in vitro, the relative expression levels of caspase 3 in protoscolices in the low, medium and high concentration groups of TA-SM in water solution groups and tablet form groups, ABZ and ABZ-SO positive control groups, medium and solvent control groups were 0.14 ± 0.01, 0.13 ± 0.01, 0.14 ± 0.01 and 0.21 ± 0.02, 0.36 ± 0.04, 0.42 ± 0.04; 0.10 ± 0.01 and 0.17 ± 0.01; 0.13 ± 0.01 and 0.10 ± 0.01, respectively, in which the TA-SM in tablet form groups were higher than the medium and solvent control groups, ABZ and ABZ-SO positive control groups (F = 58.97, 69.18, 71.81, 46.77; all P < 0.01) and the TA-SM in water solution group (t = 4.99, 10.24, 10.82; P < 0.01). The results of the in vivo experiment showed that after 60 days of treatment, the cyst inhibition rates of the tablet group and the water solution group were (24.4 ± 4.15)% and (17.2 ± 3.71)%, respectively, with a statistically significant difference between them (t = 3.15, P < 0.05). The HE staining results showed that after 60 days of treatment, the hepatocytes in the saline group were swollen, there were a large number of inflammatory cells infiltrating around, and there were a large number of vacuolar degeneration in the cytoplasm. The morphology of hepatocytes in the water solution group and the tablet group tended to be normal, the inflammatory cells were significantly reduced, and the vacuolar degeneration in the cytoplasm was reduced. SEM results showed that after 15 days of treatment, the typical vesicular structure appeared in the liver tissue lesions of mice in the saline group, while the typical vesicular structures in the liver tissue lesions of mice in the tablet group and the water solution group were significantly reduced. Conclusion TA-SM in water solution and tablet forms have certain suppressive effects on protoscolices of E. multilocularis in vitro and in vivo. The efficacy in vitro test is essentially the same as that of ABZ, and the efficacy of TA-SM in tablet form is better than that in water solution.

Polarization effect of Echinococcus granulosus antigen B on the mouse macrophage RAW264.7

JIAO Hongjie, QI Wenjing, GUO Gang, BAO Jianling, WU Chuanchuan, SONG Chuanlong, LI Jun, ZHANG Wenbao, YAN Mei

2023, 41(1):  23-28.  doi:10.12140/j.issn.1000-7423.2023.01.004

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Objective To investigate the regulatory effect of Echinococcus granulosus antigen B (AgB) on macrophage polarization. Methods After cultivated for 24 h, the RAW264.7 macrophages cells were designated to 6 groups: M1, M2, AgB, AgB+M1, AgB+M2 and blank control (M0), 3 wells each group. After all the cells attached to the well wall for 3 h, the AgB、AgB+M1、AgB+M2 group was respectively added with natural AgB extracted from sheep hydatid cyst fluid (1 000 ng/ml, final concentration); 1 h post-stimulation, the M1 and AgB+M1 group was respectively added with lipopolysaccharide (LPS, final concentration 100 ng/ml), and IFN-γ (20 ng/ml, final concentration) to stimulate differentiation for 20 h; M2 and AgB+M2 group was added with interleukin 4 (IL-4) and IL-13 (final concentration 20 ng/ml) to stimulate differentiation for 20 h; the control group was cultured in parallel without changing medium. The morphology of macrophage cells were observed microscopically. Total RNA of the macrophages in all groups was extracted for performing RT-PCR to detect the relative transcription levels of the surface markers on stimulated macrophages, including arginase 1 (Arg-1) and tumor necrosis factor α (TNF-α). The relative expression levels of Arg-1 and inducible nitric oxide synthase (iNOS) were analyzed by Western blotting. The change of IL-10 and TNF-α expression in the culture supernatant of stimulated macrophages were detected by ELISA. Results After stimulation and differentiation, most cells in the M1 group and AgB+M1 group were irregularly shaped and had antennae. The cells of M2 group and AgB+M2 group were mostly round or oval, and very few were irregular. The cells of M0 group and AgB group were partly round and oval, and partly irregular. RT-PCR showed that the relative transcription levels of Arg-1 mRNA in the M2 group and the AgB+M2 group were 189.49 ± 68.43 and 435.83 ± 123.57, respectively (t = 246.30, P < 0.01). They were higher than those in the M0 group (1.00 ± 0.00), M1 group (1.87 ± 1.29), AgB group (2.37 ± 2.06), AgB+M1 group (3.96 ± 1.92) (t = 188.50, 187.60, 187.10, 185.50, P < 0.01; t = 434.80, 434.00, 433.50, 431.90, all P < 0.01). The relative transcription levels of TNF-α mRNA in the M1 group and the AgB+M1 group were 8.34 ± 2.92 and 8.10 ± 1.54, respectively (t = 0.24, P > 0.05). They were higher than that of the M0 group (1.00 ± 0.00), M2 group (1.37 ± 0.64), AgB group (2.86 ± 0.44) and AgB+M2 group (1.62 ± 0.27) (t = 7.34, 6.97, 5.48, 6.71, P < 0.01; t = 7.10, 6.74, 5.24, 6.48, P < 0.01). Western blotting showed that the relative expression level of Arg-1 protein in the M2 group was 1.18 ± 0.35, which was higher than that in the M1 group (0.33 ± 0.18) and the AgB+M1 group (0.58 ± 0.10) (t = 0.67，0.61, P < 0.01). There was no significant difference on the Arg-1 protein relative expression level between the AgB group (1.05 ± 0.17) and the AgB+M2 group (0.97 ± 0.27) (t =0.20, 0.13, P > 0.05). There was statistical significance in AgB+M2 group compared with M1 group and AgB+M1 group (t = 0.52, 0.48, P < 0.05). The iNOS relative expression levels of M1 group and AgB+M1 group were 0.95 ± 0.21 and 0.88 ± 0.02 (t = 0.07, P > 0.05), respectively. They were higher than those in M0 group (0.03 ± 0.00), M2 group (0), AgB group (0) and AgB+M2 group (0) (t = 0.92, 0.95, 0.95, 0.95, P < 0.01; t = 0.85, 0.88, 0.88, 0.88, P < 0.01). The ELISA results showed that the expression level of IL-10 cytokine in the cell supernatant was 166.67 ± 56.67 in the AgB+M1 group and 213.33 ± 16.67 in the AgB+M2 group, respectively. They were higher than those in the M0 group (0.00 ± 0.00), M1 group (43.33 ± 36.67), M2 group (50.00 ± 43.00) and AgB group (47.50 ± 25.00) (t = 166.70, 123.30, 116.70, 119.20, all P < 0.05. t = 213.30, 170.00, 163.30, 165.80, P < 0.01). The expression levels of TNF-α cytokines in M1 group and AgB+M1 group were 833.13 ± 3.09 and 745.63 ± 118.00, respectively (t = 87.50, P > 0.05). They were higher than those in M0 group (217.50 ± 32.26), M2 group (224.69 ± 17.68), AgB group (308.44 ± 4.42), AgB+M2 group (251.25 ± 1.33) (t = 615.60, 608.40, 524.70, 581.90, P < 0.01; t = 528.10, 520.90, 437.20, 494.40, P < 0.01). Conclusion AgB can up-regulate the expression of Arg-1 in macrophages and polarize it towards M2 type, which may be an important regulatory molecule in the host-parasite immune responses, involving in the immune regulation of macrophages.

Expression and functional analysis of recombinant peptidyl-prolyl cis-trans isomerase gene of Babesia microti

SUN Jiahui, SONG Peng, CHEN Muxin, ZHOU Yan, LIN Lin, CHEN Jiaxu, CAI Yuchun

2023, 41(1):  29-35.  doi:10.12140/j.issn.1000-7423.2023.01.005

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Objective To express the recombinant peptidyl-prolyl cis-trans isomerase gene of Babesia microti (BmPPIase) in prokaryotic cells and analyze its function. Methods BmPPIase information was screened and analyzed by bioinformatics. The target fragment BmPPIase was obtained via the total gene synthesis, to construct the recombinant plasmid pET28a-BmPPIase, which was then transfected into the competent cells of Escherichia coli BL21 for prokaryotic expression. The expressed recombinant protein was analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the protein was purified via nickel column affinity. The recombinant protein concentration was estimated based on the amount of bovine serum albumin (BSA) reference standard. New Zealand rabbits were immunized with the recombinant protein four times (1.5 mg dose each on the day 1, 15, 29, and 43) at multiple sites on their back to elicit antibodies. Carotid blood was collected on the day 53 after primary immunization, to examine specific polyclonal antibody titer using enzyme-linked immunosorbent assay (ELISA), and Western blotting was used to analyze the immunogenicity of the polyclonal antibody. Further, immunofluorescence assay was used to localize its distribution in B. microti. The suppressive effect of the protein was assayed by co-incubation with mice infected blood cells in vitro, and the experiment set recombinant BmPPIase groups (added the protein 500, 250, 100, 50, and 10 μg/ml at final concentration), splicing factor 1 group (unrelated protein control, added 500, 250, 100, 50, 10 μg/ml), BSA group (100 μg/ml), and blank control. After incubation for 48 h, the culture was stained with ethidium bromide, and the proportion of infected red blood cells was measured using flow cytometry to analyze the effect of the protein on the infection of B. microti to host red blood cells. Results Bioinformatics revealed that the BmPPIase was a cyclophilin type of PPIase and was homologous to the same protein in Babesia bovis, Theileria, Blastocystis hominis, and Plasmodium. As expected, the restriction enzyme digestion of the recombinant plasmid showed that the size of the inserted gene was 531 bp, which was validated using sequencing. SDS-PAGE showed that the recombinant protein BmPPIase was a soluble protein with a relative molecular weight of 19 000. After purification and elution, the protein concentration was 1.5 mg/ml. ELISA showed that the specific polyclonal antibody titer of the recombinant protein was > 1 : 80 000. In addition, Western blotting revealed that the polyclonal antibody could specifically recognize the recombinant protein, and the immunofluorescence localization assay showed that BmPPIase was distributed on the surface of the parasite as well as secreted. In vitro inhibition experiments showed that the rBmPPIase protein at different concentrations inhibited the infection of host cells to a certain level, the relative infection rate was (47.0 ± 1.2)% at 500 μg/ml. However, with the decrease of protein concentration, the inhibition effect of BmPPIase gradually weakened, the relative infection rate was (64.0 ± 0.3)% at 250 μg/ml, (78.0 ± 1.9)% at 100 μg/ml and (82.0 ± 0.25)% at 50 μg/ml respectively. The relative infection rate was (93.0 ± 0.22)% at 10 μg/ml with no significant effect. Conclusion The recombinant protein BmPPIase is a secretory protein distributed on the parasite surface, showing significant suppressive effect on B. microti infection of host cells in vitro.

Establishment and application of a specific primers-based qPCR targeting asexual stage 18S rDNA for detecting five Plasmodium species

LI Mei, XIAO Ning, XIA Zhigui

2023, 41(1):  36-43.  doi:10.12140/j.issn.1000-7423.2023.01.006

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Objective To establish a qPCR method for detecting the 18S rDNA of the asexual stage (stage A) of 5 Plasmodium species and to evaluate its detection efficacy. Methods The 18S rDNA sequences of stage A, sporozoite stage （stage S） and oocyst (stage O) of five human malaria parasite species and the sequence from human were downloaded from GenBank and aligned to confirm the distribution pattern of the conserved and variable regions. Specific primers of Plasmodium genus were designed in the conserved regions and the specific primers of species were designed in the variable regions. Specific primer pairs were screened by the qPCR amplifying the DNA of the five Plasmodium species, Microtus babesi and Leishmania infantis as templates, and the optimal annealing extension temperature and primer concentration of the specific primer pairs were screened. The plasmids of short 18S rDNA fragments (amplified with screened primers) and long 18S rDNA fragments (amplified DNA fragments with conserved primers) of the five Plasmodium species of stage A were constructed respectively. The qPCR amplification was carried out with plasmid DNA [(5 × 10¹） - (5 × 10⁸) copy/μl] of the short fragments and plasmid DNA [(5 × 10⁴）- (5 × 10⁹) copy/μl] of the long fragment as templates, and the amplification efficiency, the minimum detection limit [including the minimum plasmid concentration and the corresponding cycle threshold (Ct) value, the coefficient of variation of repeated detection], the maximum concentration of non-target DNA when there was no amplification, and the maximum copy concentration ratio between the different Plasmodium species in the detectable mixed DNA were determined respectively. Using the DNA of the 18 blood samples infected with Plasmodium parasites (6 samples of P. falciparum, 6 samples of P. vivax, 2 samples of P. malariae, 4 samples of P. ovale, and 1 sample of P. knowlesi), which were diluted by 5 folds as the template, the selected specific primers were used for qPCR and one-step reverse transcription qPCR amplification respectively, and nested PCR amplification was performed at the same time to calculate and compare the density of the samples at the lowest detection limit. Using selected specific primers, qPCR amplification was performed on the DNA from 70 malaria patients’ blood samples (26 samples of P. falciparum, 14 samples of P. vivax, 14 samples of P. malariae, 9 samples of P. ovale, 3 samples of P. knowlesi, 4 samples of mixed infections), 65 whole blood samples of malaria-negative patients, and Amoeba sp. (1 sample), M. babesi (1 sample), and Leishmania donovani (3 samples) samples to calculate the sensitivity, specificity and conformity. Chi-square test, t-test, Mann-Whitney test or Wilcoxon rank sum test were used to compare the results of different PCR methods. Results Sequence alignment results showed that the consistency of the 18S rDNA sequence of the five Plasmodium species of parasites in stage A was 89.3%-96.7%, higher than those in stage S (60.0%-92.1%). Nine conserved regions and eight variable regions were mapped. Except for the 18S rDNA fragments of stage O of P. vivax and stage S of P. malraiae, the sequence length of the conserved regions was the same with consistency of ≥ 92%. One pair of generic primers and 5 pairs of specific species primers were selected for detecting the five species of human malaria parasites. When amplified with qPCR, the amplification efficiency of all primer pairs were 90.6%-98.8%, the optimal annealing extension temperature was 57.8 ℃, and the optimal final primer concentration was 0.3 μmol/L. The minimum detection limit for the short fragment plasmid was 5 × 10² copies/μl, the corresponding Ct value is 32.3-33.1 and the coefficient of variation of repeated detection is 0.5%-3.7%. The highest concentration of non-target DNA was (5 × 10⁶)-(5 × 10⁸) copies/μl when no amplification presented. The maximum copy concentration ratio between different Plamodium species in the mixed DNA that can be detected reached 10⁴. The average minimum paparsite density detected by one-step reverse transcription qPCR, qPCR and nested PCR were (4.1 ± 5.1), (6.0 ± 4.3) and (8.2 ± 2.9) parasites/μl respectively, and the lowest detected protozoa density was 0.01, 0.3 and 2.0 parasites/μl respectively, and the coefficient of variation were 125.6%, 72.2% and 35.8%, respectively. The sensitivity of the one-step reverse transcription qPCR is higher than those of the other two methods (Wilcoxon rank sum test, Z = -2.0, -2.0; P < 0.05). There was no statistically significant difference in the sensitivity between qPCR and nested PCR (t = -1.7, P > 0.05). The results of practical application showed that, the sensitivity and specificity of the qPCR were both 98.6%. The coincidence rate between the application results of the qPCR and the standard results was 98.6%, and their difference was not statistically significant (χ² = 0.0, P > 0.05). Conclusion The qPCR detection system using specific primers based on the 18S rDNA of Plasmodium spp. asexual stage has been established which can be applied in identifying malaria parasites in blood samples.

Surveillance of human soil-transmitted nematode infection in Anhui Province during 2016—2020

CAO Zhiguo, WANG Tianping, JIN Wei, GUO Jianduo, ZHU Lei, LIU Daohua, WANG Min, LI Qiyang, HU Mingchuang

2023, 41(1):  44-51.  doi:10.12140/j.issn.1000-7423.2023.01.007

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Objective To understand the prevalence of soil-transmitted nematode (STN) infection in Anhui Province from 2016 to 2020 to provide scientific basis for formulating the STN control strategy. Methods The surveillance for STN was performed in Anhui Province according to the National Surveillance Program for Clonorchiasis and Soil-borne Nematodiasis (provisional version) during 2016—2020. A certain number of counties (cities, districts) were selected yearly as surveillance sites, each of which was further divided into 5 sectors according to its geographic location, i.e., east, west, south, north, and central. From each sector, one township was randomly selected, and an administrative village was selected from the township to conduct surveillance. In the sampled village, no less than 200 permanent residents over 3 years old were cluster sampled. Fecal samples (> 30 g) from the enrolled residents were collected and were examined for STN eggs using the modified Kato-Katz thick smear method (two slide-reading for each sample). Children aged 3-9 were additionally examined for Enterobius vermicularis eggs with the cellophane anal swab method. The infection rate and intensity of STN were calculated and descriptively analyzed. Chi-square test and Chi-square trend test were used to compare the infection rates. Results From 2016 to 2020, 6, 16, 14, 14 and 21 surveillance sites were selected, the STN infection rate in the population in the 5 years was 2.14% (129/6 032), 0.76% (125/16 409), 1.26% (185/14 704), 0.88% (132/14 982) and 0.58% (126/21 575), respectively. The infection rate showed an overall downward trend, with a decrease of 72.90% in 2020 compared to 2016. The differences between the years were statistically significant (χ² = 143.68, P < 0.05). The infection rate of hookworm was 1.94% (117/6 032), 0.53% (87/16 409), 1.07% (158/14 704), 0.52% (78/14 982) and 0.43% (92/21 575), respectively, while that of Ascaris lumbricoides, Trichuris trichiura and E. vermicularis was relatively low (0.01%-0.24%). Among the 71 surveillance sites in the province, the highest STN infection rate was found in Qianshan City (11.02%, 103/935) in 2016, followed by Guzhen County (4.22%, 43/1 018) in 2020; and there were 19 surveillance sites with infection rate greater than 1.00%, mainly distributed in northern Anhui, accounting for 26.76% (19/71) of all the surveillance sites. From 2016 to 2020, the STN infection rate in males was 1.98% (62/3 136), 0.67% (54/8 009), 0.99% (70/7 037), 0.95% (67/7 052), 0.53% (56/10 592) and in females was 2.31% (67/2 896), 0.85% (71/8 400), 1.50% (115/7 667), 0.82% (65/7 930), 0.64% (70/10 983), respectively, there was no significant differences between genders (χ² = 0.82, 1.57, 0.73, 1.10; P > 0.05) except for 2018 (χ² = 7.54, P < 0.05). The STN infection rate showed an increasing trend with age and decreasing trend with education levels. The highest infection rate was found in the age group of "≥ 71" and the illiterate and primary school education group, which was 1.73% (11/635), 1.19% (23/1 926), 2.70% (51/1 890), 1.68% (33/1 970), 1.00% (29/2 906) and 3.08% (110/3 567), 0.99% (111/11 263), 1.63% (160/9 794), 1.14% (117/10 235), 0.61% (81/13 345), respectively. For the STN specie compositions, the composition ratio of hookworm was the highest, which was 90.70% (117/129), 69.60% (87/125), 85.41% (158/185), 59.09% (78/132) and 73.02% (92/126), respectively. For infection severety degrees, mild infection accounted for the majority, while moderate infection and severe infection were rare. Conclusion During the 13th Five-Year Plan period, the STN infection rate in Anhui Province showed a downward trend, being at a low prevalence status. Hookworm is the main infection species, and northern Anhui is the key area for STN control in the future.

Anisakis infection in marine fishes and resident awareness of anisakiasis in Qingdao in 2021

LI Dan, JI Fengying, WANG Longjiang, WANG Yongbin, LI Yuejin, YAN Ge, XU Yan, BU Xiuqin, ZHANG Benguang

2023, 41(1):  52-58.  doi:10.12140/j.issn.1000-7423.2023.01.008

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Objective To investigate the Anisakis infection in marine fishes in the Yellow Sea and local resident awareness of anisakiasis in Qingdao. Methods In October 2021, marine fishes were purchased from Fisherman’s Wharf in Qingdao and Taixi Wharf in Langya. The viscera and muscle of the marine fishes were disected to search for Anisakis larvae. The Anisakis infection in different fish species, and different organs were examined to anlyze the correlation between the infection intensity and body length and weight of the marine fishes. Five villages were randomly selected in the West Coast New District to conduct a questionnaire survey on anisakiasis knowledge among residents who were willing to participate in the survey. The knowledge awareness rate and behavior habits of the residents on Anisakis were analyzed. Surveillance for anisakiasis was conducted in local sentinel hospital. The Fisher exact test was used to compare the infection rate and the responders proportion; Spearman rank correlation analysis was performed; Chi-square test was used for analyzing the knowledge awareness rate. Results A total of 196 fish from 22 species were collected, of which 70 were infected with Anisakis. The infection rate was 35.7% (70/196). Total 13 species of marine fishes were found infected with Anisakis, accounting for 59.1% (13/22), among which Muraenesox cinereus, Pleuronichthys cornutus, Gadous macrocephaius, Scomber japonicas (all 10/10) and Nibea albiflora (9/9) were all infected with Anisakis larvae. The infection rate of Anisakis was significantly different between different species (χ² = 147.93, P < 0.01). The average infection intensity was 34.8 larvae/fish (2 439/70). The highest infection intensity was found in Gadous macrocephaius (189.3 larvae/fish), followed by Scomberomorus niphonius (25.7 larvae/fish). In Larimichthys polyactis and Pennahia argentata, Anisakis larvae were only found in mesentery and intestinal wall, accounting for 4/4 and 5/5, respectively. In Esox reicherti and Sebastiscus marmoratus, Anisakis larvae were only found in the body cavity, accounting for 13/13 and 2/2, respectively. Rank correlation analysis show that the infection intensities of Anisakis in L. polyactis and E. reicherti were positively correlated with body length (r = 0.71, 0.81, all P < 0.05) and body weight (r = 0.64, 0.80, all P < 0.05), while the infection intensity of Anisakis in Lophiomus litulon was negatively correlated with body weight (r = -0.98, P < 0.01). The infection intensity of other infected fishes was not correlated with body length and body weight. A total of 387 local residents were surveyed, 8.5% (33/387) of which had heard of anisakiasis, 4.9% (19/387) had knowledge on the harm of anisakiasis, and 7.0% (27/387) preferred to eat raw or semi-raw marine fishes, 17.8% (69/387) of residents processed raw and cooked food with shared cutting boards. The knowledge awareness rates of male and female were 11.2% (18/160) and 6.6% (15/227), respectively (χ² = 7.69, P < 0.05). The knowledge awareness rate of residents in Zhuangjiatong Village [36.7% (26/71)] was higher than that of other villages (χ² = 81.15, P < 0.01). The knowledge awareness rate in ≥ 60 years old residents was the highest, which was 11.8% (16/136), but there was no significant difference among age groups (χ² = 10.89, P > 0.05). Sentinel hospitals monitor did not find any patient infected with Anisakis. Conclusion The infection rate of Anisakis in marine fishes in Qingdao is comparatively high. The knowledge awareness rate on preventrion and control of anisakiasis in resident is low. The locals comsume raw or semi-cooked marine fishes, and process raw and cooked food with shared cutting boards.

Epidemiological trends of soil-transmitted nematode infections in Jiangxi Province, 2015—2021

CHEN Zhe, GE Jun, JIANG Weisheng, QIU Tingting, WEN Qi, ZENG Xiaojun

2023, 41(1):  59-67.  doi:10.12140/j.issn.1000-7423.2023.01.009

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Objective To understand and analyze the endemic status and transmission trends of soil-transmitted helminth (STH) infection in Jiangxi Province during 2015—2020 to provide scientific basis for formulating strategy of transmission control and interruption, and evaluate the control effect. Methods From 2015 to 2021, 13, 86, 86, 86, 86, 24 and 24 counties (cities and districts) in Jiangxi Province were selected as national surveillance sites and provincial surveillance sites. The surveillance sites were randomly selected from five geographical sections in the east, west, south, north and center, from each section, one administrative village in one township (town) was randomly selected as surveillance site. Fecal samples were collected from each enrolled participant and examined for STH eggs (two slide-reading each sample) using the modified Kato-Katz thick smear method. The infection rate and intensity of different worm species were calculated, and the infection prevalence in different eco-region was analyzed. The disposable income data of rural residents in Jiangxi Province were collected from Jiangxi Statistical Yearbook from 2015 to 2021. A questionnaire based survey was conducted to collect information on the construction of sanitary toilets in each surveillance site from the village committee from 2016 to 2021. The infection rate and constituent ratio were compared by the Chi-square test. The correlation study was conducted using Pearson correlation analysis. Results From 2015 to 2021, the infection rates of STH were 1.8% (126/7 137), 1.3% (947/71 766), 1.1% (803/70 547), 0.9% (604/69 507), 0.9% (628/69 620), 0.7% (151/21 884) and 0.9% (202/21 757) respectively, showing a decreasing trend year by year, with statistically significant differences in infection rates between years (χ² = 158.639, P < 0.01). From 2015 to 2021, the infection rates of hookworm were 1.1% (82/7 137), 0.6% (466/71 766), 0.6% (450/70 547), 0.5% (336/69 507), 0.5% (362/69 620), 0.4% (98/21 884) and 0.7% (148/21 757) respectively. From 2015 to 2021, the infection rates of Ascaris lumbricoides were 0.3% (24/7 137), 0.5% (393/71 766), 0.4% (287/70 547), 0.3% (221/69 507), 0.3% (225/69 620), 0.2% (40/21 884) and 0.2% (42/21 757) respectively. From 2015 to 2021, the infection rates of Trichuris trichiura were 0.3% (21/7 137), 0.2% (125/71 766), 0.1% (74/70 547), 0.1% (57/69 507), 0.1% (49/69 620), 0.1% (13/21 884) and 0.1% (14/21 757) respectively. The infection rates of hookworm, A. lumbricoides and T. trichiura were significantly different between years (χ² = 75.456, 112.506, 75.383, all P < 0.01). From 2015 to 2021, the infection rates were 2.0% (73/3 623), 1.3% (469/35 788), 1.1% (378/35 207), 0.8% (273/34 703), 0.9% (295/34 801), 0.6% (65/11 110) and 0.8% (88/10 620) in males and 1.5% (53/3 514), 1.3% (478/35 978), 1.2% (425/35 340), 1.0% (331/34 804), 1.0% (333/34 819), 0.8% (86/10 774) and 1.0% (114/11 137) in females. The infection rates of male and female were significantly different between years (χ² = 122.146, 49.615, P < 0.01). The infection rates of female were statistically higher than that of males in 2018 (χ² = 5.450, P < 0.05). From 2015 to 2021, the infection rates of soil-transmitted nematodes were the highest in the over-70-year-old age group, which were 3.4% (19/554), 2.3% (129/5 559), 2.1% (127/5 965), 2.0% (129/6 507), 1.7% (119/7 105), 1.3% (36/2 811) and 2.1% (68/3 177) respectively, with statistically significant differences in infection rates among the different age groups (χ² = 39.763, 150.459, 212.140, 219.129, 132.674, 43.997, 115.627, P < 0.01). From 2015 to 2021, the severely infected accounted for 1.2% (1/82), 3.9% (18/466), 1.3% (6/450), 1.5% (5/336), 0.3% (1/362), 2.0% (2/98) and 5.4% (8/148) of the total hookworm infected villagers, respectively. The differences of constituent ratio in each year were statistically significant （χ² = 22.853, P < 0.01）. Severe A. lumbricoides infection was only detected in 2016, accounting for 1.0% (4/393) of the total A. lumbricoides infection cases. T.trichiura severe infection were only detected in 2015, accounting for 4.8% (1/21) of the total T. trichiura infection cases. The infection rates of STH in 2016—2017 and 2019—2021 were the highest in Zhejiang-Fujian mountain and hill ecological regions. They were 2.3% (316/13 852), 1.5% (217/14 402), 1.4% (194/14 063), 1.2% (69/5 527) and 1.1% (60/5 535) respectively. In 2015 and 2018, the highest infection rates were 2.5% (63/2 471) and 1.2% (234/18 883) in Hunan-Jiangxi hill and mountain and Yangtze River basin ecological region, respectively. The differences in infection rates among different ecological regions in each year were statistically significant (χ² = 21.112, 156.920, 67.104, 73.315, 73.493, 37.022, 9.351, P < 0.01 or P < 0.05). From 2015 to 2021, the disposable income of rural residents in Jiangxi Province were 10 117, 11 139, 12 138, 13 242, 14 460, 15 796 and 16 981 yuan and the coverage rates of sanitary latrines in surveillance villages were 82.7% (200 994/243 000), 85.7% (256 437/299 398), 87.3% (257 553/294 951), 88.9% (250 381/281 699), 92.1% (69 428/75) and 93.8% (77 259/82 376), respectively. The infection rates of soil-transmitted nematodes in Jiangxi Province were negatively correlated with the disposable income data among the rural residents (r = -0.834, P < 0.05) while were no correlation with the coverage rate of sanitary latrines in surveillance villages (r = -0.799, P > 0.05) in Jiangxi Province during 2016—2021. Conclusion The infection rate of STH in Jiangxi Province in 2016—2021 shows an overall decline year by year, suggesting a low transmission trend. The infection rate in 41 counties has reduced to below 1.0% for 3 consecutive years.

Analysis of the causes of misdiagnosis of seven imported malaria cases in Shanghai from 2020 to 2021

ZHANG Yaoguang, JIANG Li, WANG Zhenyu, ZHU Min, ZHU Qian, MA Xiaojiang, YU Qing, Chen Jian

2023, 41(1):  68-74.  doi:10.12140/j.issn.1000-7423.2023.01.010

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Objective Comprehensive analysis of laboratory test results of the samples from imported malaria cases to determine the causes of misdiagnosis, to provide reference for reducing the occurrence of misdiagnosis. Methods A total of 112 samples of imported malaria cases reported by each district centers for disease control and prevention (CDCs) from 2020 to 2021 were collected, including blood smears, anticoagulated blood, and Plasmodium genomic DNA extracted by district CDCs (district CDC group). The samples were rechecked by the municipal malaria reference laboratories through microscopic examination, genomic DNA extraction (municipal CDC group) and multiplex PCR assay, respectively. Among the samples of microscopic positives, where as real time quantitative PCR (qPCR) negatives reported by the district CDC, additional qPCR was conducted to plot amplification curves, determine the cycle threshold (Ct) values and compare the concentration and purity of DNA between the two groups. Results Microscopic examination was performed for 112 samples and nuclear acid detection were performed for 110 samples by the district CDCs. 108 were positive samples of Plasmodium and 4 were negative samples. The microscopic concordance rate was 93.8% (105/112), as reviewed by the municipal CDC. Among them, the positive concordance rate was 94.1% (96/102) and the negative concordance rate was 9/10. A total of seven imported malaria cases were screened for misdiagnosis. Three samples had qualitative identification mistakes by the district CDCs (two P. falciparum cases were misdiagnosed as negative, and one negative case was misdiagnosed as P. falciparum). Four samples have species identification mistakes, one sample was P. malariae misdiagnosed as P. ovale, one sample was P. malariae misdiagnosed as undefined, and two samples were P. ovale misdiagnosed as P. vivax. The concordance rate of nucleic acid detection was 96.4% (106/110), as reviewed by the municipal CDC, with a positive concordance rate of 96.2% (102/106) and a negative concordance rate of 4/4. In seven misdiagnosed cases, there were no amplifications of multiplex PCR the district CDC group, and normal amplifications in the municipal CDC group. The qPCR detections were amplified normally in both district and municipal CDC groups, and the corresponding samples in the municipal CDC group had lower Ct values and the amplification curve entered the exponential growth phase earlier. The DNA concentrations in the district CDC group were all greater than those in municipal CDC group, while municipal CDC group had higher DNA purity. Conclusion Relevant district CDCs need to improve their microscopic examination capability for Plasmodium, and use appropriate DNA extraction kits and qPCR kits for Plasmodium nucleic acid detection.

Analysis of hospitalization expenses and influencing factors in surgery for patients with cystic echinococcosis in Ningxia during 2016—2021

DUAN Hongju, LI Lingjie, WU Xianglin, YAN Fang, QI Rongting, MA Tianbo

2023, 41(1):  75-80.  doi:10.12140/j.issn.1000-7423.2023.01.011

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Objective To investigate the hospitalization expenses of patients with cystic echinococcosis undergoing surgery in Ningxia for providing reference basis to reduce the economic burden hospitalized patients. Methods Data from patients who underwent cystic echinococcosis surgery from 2016 to 2021 in Ningxia were collected, including the patient’s basic information, the number of hospitalisation days and the cost of surgery and other information. SPSS 20.0 statistical software was used for statistical analysis. The average hospitalization cost was expressed in median per capita. Z test and F test were used for comparison among groups. The hospitalization expense was assigned as dependent variable. The surgery year, gender, age, occupation, education level, number of operations, hospital grade, the time interval from diagnosis to operation and the days staying in hospital were taken as independent variables for multiple linear logistic regression analysis. Result A total of 486 patients’ information undergoing surgery were collected, of them the total hospital cost, average cost per patient and the median cost was 11.391 million yuan, 23 438.7 yuan, and 20 362.5 yuan. Hospitalization expense mainly include 17 categories, the top 3 median expenses were drug fee (8 030.0 yuan), consumble material fee (5 000.0 yuan) and operation fee (3 436.4 yuan). The median hospitalization expenses of men and women were 20 126.1 yuan and 20 931.4 yuan, respectively, with no significant difference (Z = -0.552, P > 0.05). The median hospitalization expenses of patients aged 40-59 years was the highest, which was 20 835.5 yuan. The difference of hospitalization expenses among different age groups was statistically significant (Z = 8.2, P < 0.05). Among different occupational patients, the median hospitalization expense of other occupational patients was the highest, which was 22 655.0 yuan. There was no statistically significant difference in hospitalization expense of different occupational patients (Z = 1.838, P > 0.05). The median hospitalization expenses of illiterate patients was the highest, which was 23 286.4 yuan. There was no statistically significant difference in hospitalization expenses of patients with different education levels (Z = 6.729, P > 0.05). The hospitalisation cost of patients with a time interval of more than 1 year from diagnosis to surgery (20 830.1 yuan) was higher than that of patients with an interval of ≤ 1 year (19 127.0 yuan) (Z = -2.121, P < 0.05). From 2016 to 2021, the hospitalization expense of patients with one operation (20 274.3 yuan) is lower than that of patients with multiple operations (20 963.0 yuan) (Z = -2.079, P < 0.05). The median hospitalization expense of patients with hospitalization time ≤ 21 days (19 768.9 yuan) was lower than that of patients with hospitalization time > 21 days (30 732.0 yuan) (Z = 9 069.5, P < 0.01). There was no statistically significant difference between the median hospitalization expenses of patients with medical insurance (20 313.0 yuan) and those without medical insurance (29 798.5 yuan) (Z = -1.392, P > 0.05). The median hospitalization cost of patients undergoing hepatectomy was the highest (31 640.5 yuan), and the median hospitalization cost of patients undergoing different surgical methods was statistically significant (Z = 22.174, P < 0.01). From 2016 to 2021, the median hospitalization expense increased from 17 689.5 yuan to 27 090.0 yuan (Z = 27.560, P < 0.01). The median hospitalization expenses of provincial, municipal and county hospitals were 26 227.0 yuan, 16 979.0 yuan and 12 988.1 yuan, respectively. The median hospitalization expenses decreased with the decline of hospital level (Z = 146.875, P < 0.01). The average days staying in provincial and municipal hospitals is 14 days, and that in county-level hospitals is 15 days. There was a statistically significant difference in the number of days staying in hospitals of different levels (F = 9.594, P < 0.05). Multivariate logistic regression analysis showed that four factors, including operation mode, hospital level, days staying in hospital and operation year, were related to hospitalization expenses (P < 0.05). Among 486 patients, the total subsidy of the national echinococcosis project, the average cost per person and the median cost per person were 2 732 224.6 yuan, 5 621.9 yuan and 6 000.0 yuan. The total, the average capita and median of out-of-pocket expenses for each patient were 1 864 120.1 yuan, 8 176.0 yuan and 6 169.0 yuan respectively. After medical insurance and national hydatid disease project subsidies, the majority had no out-of-pocket payment accounting for 53.1% (258/486), followed by 15.2% (74/486) paid 1 000-4 999 yuan out-of-pocket and 15.2% (74/486) paid 5 000-9 999 yuan out-of-pocket. The percentage of patients with no out-of-pocket fee increased from 41.7% (45/108) in 2016 to 63.6% (35/55) in 2020 and decreased to 35.9% (23/64) in 2021. Conclusion From 2016 to 2021, the median hospitalization expenses of patients for surgery on cystic echinococcosis in Ningxia increased year by year, while the proportion of patients with no out-of-pocket fees increased year by year, but decreased to 35.9% in 2021. It is suggested that the subsidy standard of the hydatid disease prevention and control project should be appropriately raised, and patients are encouraged to choose a nearby designated hospital for treatment, so as to reduce the hospitalization costs.

TEACHING RESEARCH

Design and development of a virtual simulation project for schistosomiasis control based on the One Health concept

CHEN Lin, ZHU Jifeng, QIU Jingfan, XU Zhipeng, ZHANG Donghui, CHEN Lu, HE Jian, LI Wei, YANG Kun, JI Minjun

2023, 41(1):  81-84.  doi:10.12140/j.issn.1000-7423.2023.01.012

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Human parasitology is one of the theoretical bases for national infectious disease prevention and control, clinical diagnosis and treatment. In order to improve the undergraduate education of human parasitology, Nanjing Medical University integrated the One Health concept into the teaching of human parasitology, established the Chinese and English versions of “Schistosomiasis Prevention and Control Virtual Simulation Project”. The project is based on controlling schistosomiasis in Zanzibar aided by the Jiangsu Institute of Schistosomiasis Control and Prevention, which uses virtual simulation technology to restore the epidemic scene of African schistosomiasis. It completes the practical training of students through Internet technology. This project breaks through the limitation of time and space, simulated and trained the students’ comprehensive judgment ability and on-site response-ability, investigated the students’ theoretical mastery and practical application ability, strengthened the students’ practical thinking of parasitic disease prevention, and helped students establish the concept of One Health, which has important teaching significance. This paper introduces the design and development of this project.

REVIEWS

Research progress on the role of schistosome and its derivatives on modulation of immune dysregulatory diseases

LI Gen, SUN Tongjun, QIAN Yayun, LI Qianqian, YANG Xiaodi

2023, 41(1):  85-91.  doi:10.12140/j.issn.1000-7423.2023.01.013

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Schistosoma sp. is a parasite infect in mammals including humans. During co-evolution of host and schistosomes, the parasite have developed mechanisms that manipulate the host innate and adaptive immunity to create a microenviroments that benefi its survival. Recently, the pharmacological effects of schistosome infection and relevant molecules in the management of allergy, autoimmune and metabolic diseases has attracted great attention. This article reviews the modulatory role and underlying mechanisms of schistosomes and their derivatives in immune dysregulatory diseases, to provide a basis for subsequent laboratory and clinical investigations.

Research advances in the regulatory role of non-coding RNA in leishmaniasis

JIANG Tiange, ZENG Wenbo, LI Zhongqiu, ZHANG Yi

2023, 41(1):  92-97.  doi:10.12140/j.issn.1000-7423.2023.01.014

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Leishmania, which can parasitize in the macrophages of humans and animals, causing leishmaniasis, a neglected tropical disease that is estimated to kill up to 40 000 people each year. Non-coding RNA (ncRNA) is a genomic transcript that is not translated into proteins, and plays an important regulatory role in the occurrence and development of a variety of diseases, especially in inflammatory and immune responses. At present, there are few studies on the mechanism of action of ncRNA in leishmaniasis, and the existing studies have shown that in the early stage of Leishmania infection with the host, ncRNA plays an important regulatory role in regulating the proliferation of the parasites in the host and the establishment of infection by regulating antigen presentation, immune response, autophagy, apoptosis. This article reviews the mechanism of action of ncRNA in leishmaniasis, which is critical for the prevention, diagnosis and treatment of leishmaniasis.

Advances in parasite epitope vaccine research

WU Xiaoying, HU Yuan, CAO Jianping

2023, 41(1):  98-102.  doi:10.12140/j.issn.1000-7423.2023.01.015

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Parasitic diseases not only seriously threaten human health, but also cause huge economic losses to animal husbandry. At present, developing safe and effective vaccines is one of the strategies for the prevention and control of parasitic diseases. Epitope vaccine has the advantages of high specificity, safety, flexible design, easy production and storage, and can induce humoral and cellular immunity simultaneously. In recent years, great progress had been made in the epitope vaccines for parasitic diseases. This paper have reviewed the recent advances of vaccine epitope design and related vaccine research in parasitic diseases, in order to provide reference for the further development of effective vaccines.

SHORT COMMUNICATIONS

Difference analysis of the gut microbiome in patients with echinococcosis

CAO Deping, WU Defang, PANG Mingquan, PENG Xiaohong, LI Dayu, FAN Haining

2023, 41(1):  103-107.  doi:10.12140/j.issn.1000-7423.2023.01.016

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Fecal samples of 13 patients (7 patients with alveolar echinococcosis, 6 patients with cystic echinococcosis as infected group) and 13 healthy family members caring for patients (as control group) were collected in the Department of Hepatobiliary Surgery of Qinghai University Affiliated Hospital from October to December in 2020. The fecal DNA was extracted and the concentration was determined, and the cDNA library was synthesized by reverse transcription, and then sequenced using the metagenomic next generation sequencing. The metagenome second-generation sequencing was used to observe the changes of intestinal flora. Linear discriminant analysis was used to analyse the differences in intestinal flora between echinococcosis patients and healthy controls. The results revealed that the flora with higher relative abundance in the alveolar echinococcosis group were Alistipes onderdonkii, Bacteroides dorei, A. finegoldii, and the flora with higher relative abundance in the cystic echinococcosis group were A. shahii, Methanobrevibacter smithii, Streptococcus vestibularis. In the healthy individuals group, there were high relative abundance of B. caccae, Bifidobacterium adolescentis, Phascolarctobacterium succinatutens. Among them, A. onderdonkii, A. shahii and B. caccae are more meaningful flora. The differential flora in this study may be the intestinal microbial marker of echinococcosis, which deserves further to study.

Investigation on the insecticide resistance of Anopheles sinensis in Guizhou Province from 2018 to 2020

SHI Weifang, TIAN Zhenzao, ZHOU Jingzhu, HUANG Xueping, ZHOU Xuemei, WU Guoyan, LI Qiong, LIAO Qilang, WANG Dan

2023, 41(1):  108-111.  doi:10.12140/j.issn.1000-7423.2023.01.017

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To determine the insecticide resistance of Anopheles sinensis in Guizhou Province, A. sinensis was captured from the fields of Congjiang County, Bozhou District and Libo County of Guizhou in August 2018 to 2020. The mosquitoes were tested by the filter paper contact method, which is recommended by WHO. The susceptibility and insecticide resistance level of the captured mosquitoes to the diagnostic dose of 4% DDT, 0.05% deltamethrin and 5% malathion were evaluated. A Chi-square test was used to compare the knockdown rates after 60 min exposure and the corrected mortality rate between different study sites. After 60 min exposure, the knockdown time of the first mosquito to 0.05% deltamethrin was the longest in all study area, which takes 8.63 min, 17.52 min and 14.87 min in Congjiang, Bozhou and Libo; for 4% DDT, the knockdown time of the first mosquito was 6.33 min, 2.95 min and 11.88 min; for 5% malathion, the knockdown time of the first mosquito was 7.95 min, 2.15 min and 9.80 min, respectively. In Congjiang, Bozhou and Libo, after exposed for 60 min, the knockdown rates of tested mosquitoes to 5% malathion was the highest in all study areas, which were 83.9%, 39.0%, 59.0% respectively and there were significant differences in the knockdown rates among the three study areas (χ² = 53.98, P < 0.05). For 4% DDT, the knockdown rates were 61.9%, 33.3% and 36.2%; for 0.05% deltamethrin, the knockdown rates were 20.0%, 34.2%, 11.3%, respectively. The knockdown rates for 50% (KT₅₀) of tested mosquitoes in Congjiang, Bozhou, Libo for 4% DDT were 51.29 min, 127.65 min and 80.05 min; for 0.05% deltamethrin were 208.27 min, 74.69 min and 420.14 min; for 5% malathion were 29.33 min, 75.60 min and 42.19 min, respectively. The adjusted mortality rates of tested mosquitoes in Congjiang, Bozhou, Libo for 5% malathion were 90.1%, 95.0% and 95.9% respectively, which suggested the existence of resistance. There were no significant differences among the three study areas (χ² = 4.64，P > 0.05). For 4% DDT, the mortality rates were 65.6%, 38.6% and 55.0%; for 0.05% deltamethrin, the mortalityrates were 35.2%, 35.2% and 10.4%, respectively, which suggested that all tested mosquitoes had developed high level of resistance. This study suggested that An. sinensis in Guizhou Province have developed high level of resistance to deltamethrin, and maintained resistance to DDT and may have possible resistance to malathion.

Prevalence of trematode metacercariae in Pseudorasbora parva and species identification in Qiqihaer area

ZHANG Fengyu, LIU Liu, ZHANG Jing, ZHANG Hao

2023, 41(1):  112-116.  doi:10.12140/j.issn.1000-7423.2023.01.018

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To understand the prevalence of trematode metacercariae and their species in Pseudorasbora parva from Nenjiang River around Qiqihaer City. P. parvas were collected from Nenjiang River around Qiqihaer City at the begining of each month from June to November in 2021. The prevalence of metacercaria of trematodes was examined by direct tablet, and the metacercaria of trematodes were isolated and purified by digestion. The metacercaria of trematodes were preliminarily classified by morphology. Mice (50-80 metacercaria/mouse) aged 6 weeks and ducks (80-100 metacercaria/duck) aged 2 weeks were fed with different types of metacercaria. The mice and ducks were dissected on the 30th and 25th day after infection, and the adult worms in the hepatobiliary duct were collected and stained with magenta acetate for morphological identification. The prevalence of metacercaria infection in P. parva was analyzed by month and season. SPSS 26.0 software was used for statistical analysis, and the prevalence was compared by χ² test. From June to November 2021, a total of 600 P. parva were collected, and 371 were infected with metacercaria, with an prevalence of 61.8%. A total of three different type metacercaria were obtained. According to the morphology, it was preliminarily identified as C. sinensis, M. orientalis, M. taiwanensis metacercaria, the prevalences were 47.7%(286/600), 15.5% (93/600) and 23.7% (142/600), respectively. The adult worms obtained from mice and ducks infected with the three types of metacercaria of trematodes were identified by morphology as C. sinensis, M. orientalis and M. taiwanensis. From June to November, the prevalences of C. sinensis metacercaria in P. parva were 26.0% (26/100), 31.0% (31/100), 52.0% (52/100), 66.0% (66/100), 63.0% (63/100) and 48.0% (48/100), respectively. The prevalences of M. orientalis metacercaria were 5.0% (5/100), 11.0% (11/100), 15.0% (15/100), 17.0% (17/100), 26.0% (26/100) and 19.0% (19/100), respectively. The prevalences of M. taiwanensis metacercaria were 17.0% (17/100), 15.0% (15/100) 19.0% (19/100), 23.0% (23/100), 32.0% (32/100) and 36.0% (36/100), the prevalences of the three metacercaria types in different months have statistical significance (χ² = 53.61, 19.51, 20.11; P < 0.05). The prevalences of metacercaria of C. sinensis, M. orientalis and M. taiwanensis in summer and autumn were 36.3% (109/300), 59.0% (177/300); 10.3% (31/300), 20.7% (62/300); 17.0% (51/300) and 30.3% (91/300), respectively, there were statistically significant differences in the prevalence of the three metacercaria in summer and autumn (χ² = 30.89, 12.23, 14.71; all P < 0.05). The difference of prevalences between three kinds of metacercaria in summer and autumn was statistically significant (χ² = 22.29, 33.96; all P < 0.05). P. parvas were infected with metacercaria of C. sinensis, M. orientalis, M. taiwanensis in the Qiqihar section of the Nenjiang River. The prevalence of three types of metacercaria were the highest in September, October and November respectively.

Surveillance of resistance of Anopheles sinensis in some areas of Henan Province

HE Zhiquan, HU Yabo, WANG Dan, ZHANG Hongwei, LIU Ying, YANG Chengyun, QIAN Dan, JI Penghui, JIANG Tiantian, LU Deling

2023, 41(1):  117-120.  doi:10.12140/j.issn.1000-7423.2023.01.019

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To understand the resistance of Anopheles sinensis to insecticides, An. sinensis were captured near sheepfolds, pigstys or cowsheds in four counties (districts), including Pingqiao District of Xinyang City, Xiangfu District of Kaifeng City, Xiangcheng District of Xuchang City, and Tanghe County of Nanyang City in Henan Province from July to September 2021. The An. sinesis was identified by morphology. The insecticide susceptibility test of female An. sinensis was determined by the filter-paper bioassay, which is recommended by WHO, using 0.05% deltamethrin (0.017 8 g/m²), 0.15% beta-cyfluthrin (0.053 4 g/m²), 0.1% propoxur (0.035 7 g/m²), and 5% malathion (1.78 g/m²). The result showed that a total of 1 334 An. sinensis were used for insecticide susceptibility test in Pingqiao District, Xiangfu District, Xiangcheng County and Tanghe County. The mosquitoes were exposed to the four insecticide films, including, beta-cyfluthrin, deltamethrin, propoxur and malathion. The longest knockdown time of the first mosquito exposed to deltamethrin was seen in mosquitoes collected from Tanghe was 25.85 min, and the shortest knockdown time of the first mosquito exposed to deltamethrin was seen in mosquitoes collected from Pingqiao, which was 2.83 min. The knockdown rate of cypermethrin against the tested mosquitoes from Pingqiao District, Xiangfu District, Xiangcheng County and Tanghe County after 60 min was 24.42% (21/86), 2.47% (2/81), 37.31% (25/67) and 45.45% (25/55), respectively (χ² = 39.355, P < 0.05）. The knockdown rate of deltamethrin against mosquitoes from all different sites were 75.86% (66/87), 2.33% (2/86), 10.81% (8/74) and 20.35% (23/113), respectively (χ² = 142.630, P < 0.05). The knockdown rate of propoxur against mosquitoes from all different sites were 48.28% (42/87), 22.62% (19/84), 50.50% (51/101) and 72.34% (34/47), respectively (χ² = 32.635, P < 0.05). The knockdown rate of malathion against mosquitoes from all different sites were 66.29% (59/89), 15.69% (16/102), 82.54% (104/126) and 91.84% (45/49), respectively (χ² = 133.489, P < 0.05). The mortality rate of mosquitoes from Tanghe exposed to beta-cypermethrin and propoxur for 24 h was 70.91% and 80.60%, respectively, and the mortality rate of mosquitoes from Pingqiao exposed to deltamethrin for 24 h was 85.85%, which had reached resistance. The 24 h mortality rates of mosquitoes from Pingqiao and Tanghe to malathion were 97.43% and 97.67%. The resistance levels were possible resistance; The 24 h mortality rates of mosquitoes from Xiangfu and Xiangcheng to malathion were 64.23% and 99.21%. The resistance levels were resistance and susceptible. The resistance level of An. sinensis to malathion varies from susceptible to resistance but has developed resistance to deltamethrin, beta-cyfluthrin, and propoxur in four surveillance sites of Henan Province.

Analysis of Blastocystis hominis infection in long-term hospitalized patients in Tianmen City

ZHU Mingchao, ZHU Ya, ZHAO Jianzhong, YUAN Huizhen

2023, 41(1):  121-124.  doi:10.12140/j.issn.1000-7423.2023.01.020

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Basic information (gender, age, fecal characteristics, occult-blood test results, clinical signs and symptoms, etc) of long-term hospitalized patients in Tianmen First People’s Hospital during June—September 2021 collected. The fecal samples of patients were collected and examined under microscope by direct smear with normal saline. The DNA were extracted from the suspected Blastocystis hominis positive fecal samples. Specific primers of B. hominis BhRDr and RD5 were used to amplify the target bands, and the positive fecal samples were sent to Norsay Genomics Research Center for sequencing. Giardia lamblia and fragile bicarbonate amoeba infection were also observed in the fecal samples of the patients. Anticoagulant whole blood and non-anticoagulant venous blood were collected for blood analysis. Among the 857 patients, 42 cases were positive for B. hominis, with a positive rate of 4.90%. The positive rate of males was 4.27% (19/445) and that among the females was 5.58% (23/412). There was no significant difference between the gender groups (χ² = 0.791, P < 0.05). In different age groups, the positive rate was the highest in the ≥ 61 years old group (6.38%, 22/345). There was no significant difference in positive rates among different age groups (χ² = 2.700, P > 0.05). Among different fecal characters, the positive rate of the loose stool samples was the highest (18.37%, 27/147) (χ² = 98.930, P < 0.05). The positive rate was 6.72% (9/134) in patients with hematochezia and 4.56% (33/723) in patients without hematochezia, with no significant difference (χ² = 1.123, P > 0.05). Among different clinical symptoms, patients with abdominal pain and diarrhea had the highest positive rate (4/11) (χ² = 53.956, P < 0.05). Six patients were infected with B. hominis mixed with G. Lamblia, and two were infected with amoeba fragile. The red blood cell count and hemoglobin of B. hominis infected patients were (3.97 ± 7.39) × 10¹²/L and (119.02 ± 21.49) g/L, both of which were lower than that of the non-infected patients [(4.27 ± 0.44) × 10¹²/L, (126.52 ± 14.30) g/L] (t = 2.556, 2.235; P < 0.05). There were four genotypes of B. hominis (ST1, ST3, ST6 and ST7), and ST3 was the dominant one, accounting for 57.1% (24/42).

CASE REPORTS

A case of human Fasciola hepatica infection

LU Shuihuan, LUO Yanying, CAO Lijun, ZHOU Xianbao, HUANG Yingwen, HE Xueqiang

2023, 41(1):  125-127.  doi:10.12140/j.issn.1000-7423.2023.01.021

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A 66-year-old female patient, who was a farmer, went to the local county hospital on February 15, 2022 for treatment due to upper abdominal pain and nausea for 7 days. The abdominal CT examination showed dilatation of the common bile duct, intrahepatic bile duct stones in the left lobe of the liver and dilatation of the left lobe of the bile duct, and atrophy of the left lobe of the liver. She was transferred to the 924 Hospital of the Joint Logistics Support Force of the Chinese People’s Liberation Army in the following day. The patient lived in the mountain area of Guilin, Guangxi and had history of drinking uncooked mountain spring water and ate raw Houttuynia cordata for a long time. There was positive tenderness but no rebound pain under the xiphoid process at admission examination. Laboratory test showed normality in routine blood test. Abdominal enhanced MRI examination showed that the left lobe of liver was atrophied, the intrahepatic bile duct in the left lobe of liver and common bile duct were significantly dilated, and granular or strip T2 low signal shadow was seen in the bile duct. After excluding the relevant surgical contraindications, endoscopic retrograde cholangiopancreatography was performed. During the operation, a viable Fasciola hepatica was removed with a basket, and the nasal bile duct was retained for drainage. The worm was transferred to the Department of Human Parasitology in Basic Medical College of Guilin Medical College and identified as Fasciola hepatica. After the operation, 0.6 g of albendazole and 0.6 g of praziquantel were given orally every 12 hours for 2 days. After two months of operation, the patient had no symtoms. CT examination showed atrophy of the left lobe of the liver and dilation of intrahepatic bile duct in the left lobe of the liver and common bile duct.

A case of atypical cerebral cysticercosis

DUAN Yueli, YU Yiqi, ZHU Haoxiang, WANG Xinyu

2023, 41(1):  128-130.  doi:10.12140/j.issn.1000-7423.2023.01.022

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In October 2021, a 55-year-old female patient from Bengbu, Anhui, was admitted to Huashan Hospital Affiliated to Fudan University with the main complaint of “intermittent low fever, dry cough with hydrocephalus for more than 1 year”. In the previous year, the patient repeatedly developed fever with a body temperature of 37.5-38.0 ℃ and paroxyresis of dry cough. Head CT examination revealed hydrocephalus, and laparoscopic-assisted ventriculoperitoneal shunt was performed in the hospital on September 28. The fever and cough remained after the operation, and enhanced head MRI showed “abnormal enhancement shadows in the sellar region and basal cistern”. A total number of 208 721 copies of Taenia solium sequence was detected in the cerebrospinal fluid based on metagenomic next-generation sequencing (mNGS), and the serum cysticercosis antibody was positive. In the epidemiological history, the patient denied eating raw pork, but had eaten incompletely cleaned raw vegetables for many times in the past 2 years. Combining imaging findings, laboratory tests and mNGS results, the patient was diagnosed with “cerebral cysticercosis”. The patient then started to take albendazole for two courses, the total amount of each course was 150 mg/kg, the first course lasted for 12 days, and the second course lasted for 8 days, and the patient had no noticeable discomfort during the treatment. After the first course of treatment, enhanced head MRI showed abnormal enhancement shadow in the saddle region and basal cistern, and the enhancement degree was less than that of the previous film. The re-examination of the head CT showed hydrocephalus, changes after the lateral ventricular drainage, the extent of ventricular enlargement had slightly reduced, and the density of the saddle area was not uniform. Regular follow-up was undertaken at the end of the course of treatment by other hospitals.