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30 June 2023, Volume 41 Issue 3

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EXPERT VIEWPOINT

The control strategies for zoonoses under climate change based on the One Health concept

CHAO Anqi, LI Huimin, HU Qinqin, ZHOU Xiaonong, GUO Xiaokui, YIN Kun

2023, 41(3):  263-269.  doi:10.12140/j.issn.1000-7423.2023.03.001

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Zoonotic diseases pose a serious threat to human health and ecological security. Climate change facilitates the crossover and spread of zoonotic diseases by affecting pathogens, hosts, vectors, and human activities, therefore threatening global public health. This article summarizes the impact of climate change on the spread of zoonotic diseases and explores the effective strategies based on the One Health approach to protect human health and safety.

ORIGINAL ARTICLES

Transcriptome analysis of mice brain chronically infected with Toxoplasma gondii and validation of the kynurenine pathway associated with depression

ZHANG Chi, CHEN Jiating, XIN Zixuan, YANG Lili, YANG Zihan, PENG Hongjuan

2023, 41(3):  270-278.  doi:10.12140/j.issn.1000-7423.2023.03.002

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Objective To screen the differentially expressed genes (DEGs) by comparing the transcriptome of the brain tissues between the mice chronically infected with Toxoplasma gondii and normal mice, to analyze the relative transcription level of DEGs in the depression-related kynurenine (KYN) pathway and to provide a theoretical basis for exploring the mechanism of depression-like symptoms caused by Toxoplasma gondii chronic infecttion in mice. Methods SV129 male mice (n = 18) were randomly and equally divided into the infection group and the control group. Mice in the infection group were intraperitoneally injected with 120 tachyzoites of T. gondii ME49 strain (200 μl), and mice in the control group were injected with the same volume of PBS. After 3 months post-infection, mice brain tissues of the two groups were collected for extraction of total RNA to undertake transcriptome sequencing for screening DEG. With the DEGs obtained, cluster analysis, gene ontology (GO) functional annotation analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) functional annotation and enrichment analysis were performed. Eight DEGs [interferon-γ (IFN-γ), indoleamine 2,3-dioxygenase 1 (IDO1), IDO2, kynurenine-3-monooxygenase (KYNU), kynurenine-3-monooxygenase (KMO), 3-hydroxyanthranilate 3,4-dioxygenase (3-HAO), vimentin (Vim) and brain-derived neurotrophic factor (BDNF)] related to KYN pathway associated with depression were selected to examine each gene’s relative transcription level by quantitative real-time PCR (qRT-PCR), using glyceraldehyde-3-phosphate dehydrogenase gene as an internal reference. Results Transcriptome sequencing found 2 295 DEGs in the brain of the mice from the infection and control groups, of which 2 016 were up-regulated and 279 were down-regulated. GO analysis showed that localisation was the most significantly enriched biological process, with a total of 257 DEGs. The most significantly enriched in cellular components was the protein-containing complex, with a total of 425 DEGs. The most significantly enriched molecular function was molecular transducer activity, with 177 DEGs. The largest number of DEGs enriched in biological process, cell component and molecular function were cell process, cell part and binding, with 1 039, 1 240 and 1 088 DEGs, respectively. KEGG analysis showed that the top three up-regulated metabolic pathways were the immune system, signaling transduction, and viral infectious disease, and the top three down-regulated pathways were signal transduction, signaling molecules and interaction and immune system. Functional enrichment analysis showed that 77 pathways were significantly enriched. The signaling pathways related to depression include tumor necrosis factor signaling pathway, neuroactive ligand-receptor interaction, NF-kappa B signaling pathway, JAK-STAT signaling pathway, necroptosis, apoptosis, chemokine signaling pathway, KYN pathway. The qRT-PCR results showed that the relative transcription levels of IFN-γ, IDO1, IDO2, KYNU, KMO, 3-HAO and Vim genes in the infection group were 3 023.08%, 355.52%, 190.17%, 496.55%, 339.92%, 212.74% and 507.34%, if the relative transcript level of control mice was taken as 100%. Compared with the control group, the transcription was significantly up-regulated (t = 3.782, 3.749, 3.226, 2.908, 2.533, 5.656, 2.948; all P < 0.05 or 0.01). The relative transcription level of BDNF was 63.32%, which was significantly down-regulated (t = 2.398, P < 0.05). The fold change of IFN-γ, IDO1, IDO2, KYNU, KMO, 3-HAO, BDNF, Vim obtained by qRT-PCR was 4.96, 1.74, 0.89, 2.10, 1.60, 1.06, -0.94, 2.18, respectively. The fold change obtained by transcriptome sequencing was 7.30, 0.55, 0.80, 3.83, 2.75, 3.53, -0.86 and 1.93, respectively. The transcriptional trend obtained by qRT-PCR was consistent with that obtained by transcriptome sequencing. Conclusion DEGs from brain tissues of mice chronically infected with T. gondii were screened. Transcriptome analysis revealed that the immune response of central nervous system of the mice with chronic infection of T. gondii was continuously activated. Seven DEGs in KYN pathway related to depression showed up-regulated transcription level.

Effect of locking galectin-receptor interaction on the immunopathology of small intestine of Toxoplasma gondii-infected mice

OU Yangran, LIU Xingzhuo, HUANG Shiguang, LYU Fangli

2023, 41(3):  279-285.  doi:10.12140/j.issn.1000-7423.2023.03.003

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Objective To investigate the regulatory effect of galectin-receptor interaction on the small intestine immunopathology of Toxoplasma gondii-infected mice. Methods Eighteen female BALB/c mice were randomly divided into 4 groups: 4 mice in uninfected group (naive group), 4 mice in lactose group (lactose group), 5 mice in T. gondii infection group (Tg group) and 5 mice in T. gondii infection + lactose group (Tg+lactose group). Each mouse in the Tg group and the Tg+lactose group was intraperitoneally (i.p.) injected with 1 000 tachyzoites of T. gondii RH strain, while the naive group and lactose group were i.p. injected with 0.2 ml PBS. Starting from day 0 post infection, each mouse in the Tg+lactose group and the lactose group was i.p. injected with 0.2 ml 0.2 mol/L of lactose, while each mouse in the naive group and the Tg group was i.p. injected with an equal volume of PBS, once in the morning and once in the evening for 7 consecutive days. After infection with T. gondii, the mice survival time in each group was recorded. The mice were euthanized on the 7th day after infection to collect middle segment of jejunum from each mouse for prepareing paraffin sections, which were stained with hematoxylin and eosin (HE) to observe the pathological changes; from the lower segment of jejunum of each mouse, total RNA was extracted and reverse-transcribed, and used in quantitative real-time reverse transcription PCR (qRT-PCR) with β-actin as an internal reference gene to detect the relative mRNA expression level of surface antigen 1 (SAG1), galectin-3, galectin-9, T cell immunoglobulin mucin 3 (Tim-3), leukocyte differentiation antigen 137 (CD137), interleukin 12 (IL-12), interferon-γ (IFN-γ), IL-10, IL-4, transforming growth factor β (TGF-β), chemokine receptor 2 (CCR2) and chitinase 3 like molecule 3 (Ym1). Results After infection with T. gondii, there was no mice died in the naive group and the lactose group. The survival time of the mice in the Tg group was 182-188 h, and the survival time of the mice in the Tg+lactose group was 180-182 h; the difference of the survival time between the two groups was statistically significant (χ² = 19.52, P < 0.05). HE staining showed no inflammation in the mice jejunal tissue in the naive group and lactose group. Shortened intestinal villus, shallower intestinal crypts, necrosis of epithelial cells at the top of villi and inflammatory cell infiltration in the intestinal mucosa were observed in the mice small intestine from the Tg group and Tg+lactose group. Compared with the Tg group, the pathological change of the mice small intestine in the Tg+lactose group was more severe. The qRT-PCR results showed that the relative mRNA expression of SAG1 in the mice small intestine of the Tg+lactose group was 9.17 ± 1.65, which was higher than that in the Tg group (1.00 ± 0.84, t = 4.40, P < 0.05). The relative mRNA expression levels of galectin-3 in the mce small intestine of the naive group, lactose group, Tg group, and Tg+lactose group were 1.00 ± 0.28, 1.71 ± 0.31, 2.46 ± 1.11, and 7.10 ± 1.57, respectively (F = 10.15, P < 0.01). The mRNA expression levels of galectin-9 in the 4 groups were 1.00 ± 0.31, 1.44 ± 0.26, 3.21 ± 1.01, and 7.00 ± 1.08, respectively (F = 14.53, P < 0.01). The mRNA expression levels of Tim-3 in the 4 groups were 1.00 ± 0.12, 0.88 ± 0.28, 1.64 ± 0.31, and 4.89 ± 0.69, respectively (F = 19.15, P < 0.01). The mRNA expression levels of CD137 in the 4 groups were 1.00 ± 0.42, 1.03 ± 0.30, 0.89 ± 0.11, and 3.84 ± 0.77, respectively (F = 8.46, P < 0.01). The mRNA expression levels of IL-12 in the 4 groups were 1.00 ± 0.35, 1.14 ± 0.56, 12.37 ± 4.43, and 18.42 ± 3.89, respectively (F = 10.18, P < 0.01). The mRNA expression levels of IFN-γ in the 4 groups were 1.00 ± 0.56, 1.65 ± 0.53, 5.57 ± 1.84, and 21.26 ± 6.48, respectively (F = 10.38, P < 0.01). The mRNA expression levels of IL-10 in the 4 groups were 1.00 ± 0.20, 1.10 ± 0.25, 8.65 ± 2.52, and 21.98 ± 3.96, respectively (F = 20.84, P < 0.01). The mRNA expression levels of IL-4, TGF-β, and CCR2 in the mice small intestine among the naive group, lactose group, Tg group, and Tg+lactose group had no statistically significant differences (F = 1.09, 4.74, and 2.03, P > 0.05). Ym1 mRNA expression was not detected in the naive group and lactose group, and Ym1 mRNA expression levels between the Tg group and the Tg+lactose group had no statistically significant difference (t = 0.24, P > 0.05). Conclusion Blockage of galectins-receptor interaction in mice infected with T. gondii leads to increased parasite burden in small intesting tissues, and aggravated pathological impairment, as well as upregulated expression of galectin-3, galectin-9, Tim-3, CD137, IL-10 and IFN-γ.

Effect of excretory-secretory antigen TPx of Cysticercus cellulosae on activation of dendritic cells in piglets

YE Jingming, HE Wei, LIU Huiyuan, YU Xiao, LUO Bo, LIU Meichen, ZHOU Biying

2023, 41(3):  286-293.  doi:10.12140/j.issn.1000-7423.2023.03.004

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Objective This study investigates the effect of thioredoxin peroxidase (TPx) in the excretory-secretory antigen (ESA) of Cysticercus cellulosae on activation of dendritic cell (DC) in piglets. Methods Healthy piglet medulla-derived DC were cultured in vitroo, in which lipopolysaccharide (LPS) was added at a final concentration of 100 ng/ml on 7 d for stimulation for 2 days, and then continuously cultured for 2 more days to collect immature DC (imDC) and mature DC (mDC) separately. The morphological changes of DCs were observed by light microscopy and scanning electron microscopy on 1 to 9 d of culture. The expression of surface markers CD1 and major histocompatibility complex (MHC-Ⅱ) was detected by flow cytometry. The 7 d imDC was used in the assay with the assigned groups of negative control, TPx, ESA and LPS positive control, to which RPMI 1640 medium, TPx (50 μg/ml), ESA (50 μg/ml) and LPS (100 ng/ml) was added, respectively, to stimulate for 48 h for examining the expression of DC surface markers MHC-Ⅱ, CD80 and CD86 using flow cytometry and for detecting secretion levels of tumor necrosis factor-α (TNF-α), interleukin (IL-6), IL-10, IL-12 in DC culture supernatant by ELISA. One-way ANOVA was used for comparison between multiple groups, and LSD-t test was used for two-way comparison between groups. Results Under ligth microscope, imDC were ovoid in shape with single form at the first day of culture; with the extension of culture time, DC increased in size, appeared pseudopods and spines and other features, and changed from ovoid to irregular shape. Scanning electron microscopy showed that compared with imDC, mDC had irregular morphology, roughly long shuttle shape, and numerous protrusions of different lengths radiating from the cytosol, which were distributed in a dendritic pattern, a typical dendritic structure. Flow cytometry showed that the expression of CD1 and MHC-Ⅱ in imDC was (0.113 ± 0.005)% and (0.430 ± 0.016)%, respectively, which was lower than that of mDC (21.400 ± 0.327)% and (21.333 ± 0.450)% (t = 130.341, 92.906, both P < 0.05). The expression levels of MHC-Ⅱ, CD80, and CD86 in the TPx group were (15.300 ± 0.245)%, (22.900 ± 0.374)% and (13.033 ± 0.249)%, respectively, which were lower than those in the LPS positive control group (19.000 ± 0.374)%, (31.600 ± 0.082)%, and (21.300 ± 0.245)% (t = 11.53, 46.32, 43.84, all P < 0.05) and the ESA group (18.365 ± 0.618)%, (40.400 ± 0.356)% and (30.300 ± 0.283)%] (t = 9.55, 93.17, 91.57, all P < 0.05). The MHC-Ⅱ expression level in the TPx group was higher than that of the negative control group (12.133 ± 0.492)% (t = 9.87, P < 0.05). ELISA results showed that IL-6 level in DC of the TPx group was 15.682 ± 0.660, which was ower than that of 21.041 ± 0.901 in the control group (t = 6.51, P < 0.05); TNF-α (35.711 ± 4.196), IL-6, IL-10 (22.216 ± 1.357) and IL-12 (16.799 ± 0.523) were all lower than those of the LPS positive control group 169.037 ± 7.823, 42.118 ± 1.932, 34.730 ± 1.772, 52.504 ± 2.431 (t = 36.79, 32.09, 13.09, 35.05, all P < 0.05); IL-12 level were lower than that of the ESA group at 23.854 ± 1.020 (t = 6.93, P < 0.05). Conclusion TPx mediates immune tolerance by inducing high expression of DC surface molecules MHC-Ⅱ, low expression of CD80 and CD86, and reducing the secretion levels of IL-6.

Establishment and application of a rolling circle amplification method based on novel-miR1 for detection of Cysticercus pisiformis infection in rabbit

CHEN Guoliang, WANG Liqun, LI Yanping, LIU Tingli, LI Hong, ZHANG Shaohua, LUO Xuenong, QIANG Wenjun

2023, 41(3):  294-299.  doi:10.12140/j.issn.1000-7423.2023.03.005

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Objective To establish a rolling circle amplification (RCA) method for detection of Cysticercus pisiformis infection in rabbits. Methods The novel-miR1 derived from C. pisiformis in rabbit serum served as the diagnostic target to establish an RCA diagnostic method for cysticercosis pisiformis fection in rabbit. To establish the RCA method, ligation sequence and the locking probe sequence were designed, and five important reaction conditions were optimized, including the ratio of ligation sequence and padlock probe, ligation reaction time, ligase dosage, amplification reaction time, and dNTP dosage. The sensitivity of the RCA method was assessed, the novel-miR1 standard was serially diluted into 9 samples of different concentrations ranging from 1 fmol/L to 100 nmol/L. The sensitivity and specificity of the optimized RCA methods were assayed using the serum miRNA from 20 healthy rabbits and 20 C. pisiformis infected rabbits (Laboratory preserved samples), and analyzed by the receiver operating characteristic curve (ROC curve) method. To evaluate the application effectiveness of the RCA, 20 female rabbits were infected by gavage with 1 000 eggs of T. pisiformis for collection of serum every month post-infection, which were used to prepare miRNA for application in RCA method under optimized condition. Results The agarose gel electrophoresis results showed that the RCA amplification products remained in the sampling wells of agarose gel, forming a bright band. The tests to optimize RCA condition indicated that the concentration of ligation sequence was 2 μmol/L, padlock probe was 1 μmol/L (ratio of ligation sequence and padlock probe was 2∶1), T4 DNA ligase was 350 U, and the efficacy of ligation reaction was found the highest when ligating for 180 min. The optimal amplification reaction system was 0.5 μmol/L dNTP and amplification reacted 240 min in 100 μl amplification reaction system. Lastly, the RCA method limit of detection was proved to be 10 pmol/L. The RCA method detected showed that the average serum miRNA fluorescence intensity of the samples from healthy rabbits and C. pisiformis infected rabbits were 53.298 ± 1.707 and 97.498 ± 5.892, respectively, which was statistically significant (t = 7.206, P < 0.01). ROC curve showed that the RCA method cut-off value was 61.69 and both sensitivity and specificity were 95%, the area under the curve (AUC) is 0.955 0, likelihood ratio was 19.00. Using the RCA method to detect 20 healthy rabbits’ serum miRNA found 19 samples were negative and 1 positive. The results of RCA detection of the C. pisiformis infected rabbits serum miRNA at different times found that 17 were positive, 2 were suspected, 1 were negative 1 month post-infection. One was negative, and 19 were positive 2 mouths post-infection. Three were negative, and 17 were positive 3 months post-infection. Conclusion An RCA method was established for detecting C. pisiformis infection in rabbits. It was demonstrated that novel-miR1 could be detected by RCA in the serum of rabbits within 3 months post-infection of C. pisiformis, showing good application potential.

Preparation of Echinococcus granulosus peptide embedded in chitosan quaternary ammonium salt nanoparticles

WU Xiaoying, HU Yuan, CAO Jianping

2023, 41(3):  300-305.  doi:10.12140/j.issn.1000-7423.2023.03.006

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Objective To prepare nanoparticles (NPs) containing the epitope peptide FL46 of Echinococcus granulosus using hydroxypropyltrimethyl ammonium chloride chitosan (HACC) and sodium tripolyphosphate (TPP) as carriers, to screen the optimal preparation conditions, and evaluate the encapsulation effect and morphological characterisation. Methods The FL46-HACC-NPs suspension was prepared by slowly adding FL46 solution dropwise to HACC solution using the ionic cross-linking method, stirring at 800 r/min and adding TPP solution dropwise. The average particle size, polydispersity index (PDI) and surface potential (Zeta) were measured using a nanoparticle size potentiometer, the microscopic morphology was observed under the transmission electron microscope and the encapsulation rate was calculated by BCA protein concentration measurement assay kit. A three-factor, four-level orthogonal experiment was conducted with HACC concentration (0.25, 0.50, 0.75, 1.00 mg/ml), TPP concentration (0.25, 0.50, 0.75, 1.00 mg/ml), and HACC to TPP mass ratio (8∶1, 9∶1, 10∶1, 11∶1), with single factor method to examine the effect on the characterisation of FL46-HACC-NPs. Results FL46-HACC-NPs solution can be clarified, milky and precipitated at different mass concentrations and mass ratios of HACC and TPP. After single-factor experiments, the optimal preparation conditions were screened for small particle size and high encapsulation rate. The optimal preparation conditions were HACC concentration of 0.25 mg/ml, TPP concentration of 0.50 mg/ml, and the mass ratio of HACC to TPP 9∶1. Under the optimal conditions, spherical or polygonal FL46-HACC-NPs with the particle size of (307.37 ± 2.17) nm, PDI of (0.228 ± 0.008), Zeta potential of (29.77 ± 0.46) mV and encapsulation rate of (75.95 ± 1.60) % can be prepared. Transmission electron microscopy shows that the FL46-HACC-NPs have a homogeneous morphological structure and are sphere-like or polygonal in shape. Conclusion Optimised FL46-HACC-NPs can be effectively prepared by the ion cross-linking method, which can provide basis for the study of the protective effect of E. granulosus polypeptide vaccine.

Sequence analysis of mitochondrial co1 and nd1 genes in Echinococcus granulosus in Yunnan Province

LI Benfu, WANG Zhengqing, XU Qian, ZI Jinrong, YAN Xinliu, PENG Jia, LI Jianxiong, CAI Xuan, WU Fangwei, YANG Yaming

2023, 41(3):  306-311.  doi:10.12140/j.issn.1000-7423.2023.03.007

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Objective To analyze the genotype and sequence polymorphisms of mitochondrial cytochrome oxidase 1（co1）and NADPH dehydrogenase subunit 1 (nd1) in Echinococcus granulosus from Yunnan Province. Methods Echinococcus of animal origin was collected from cysts isolated from liver or lung lesions of cattle, sheep and free-range pigs at slaughterhouses in Shangri-La, Daguan, Eryuan, Lushui and Weixi counties of Yunnan Province. Echinococcus of human origin was obtained from the focal tissues surgically removed from hospitals in Jianchuan, Yunlong, Longyang and Yulong Counties, then the E. granulosus was selected after pathogenic identification, and used to extract genomic DNA with a DNA extraction kit, followed by PCR amplification and sequencing of co1 and nd1 genes. The sequence homology and single nucleotide polymorphisms were analyzed using BLAST aligment. Phylogenetic trees based on co1 and nd1 were constructed by the neighbor-joining method using MEGA-X software. Results A total of 62 samples of Echinococcus were collected, 36 of which were pathogenically confirmed as E. granulosus. A total of 32 gene co1 sequences were successfully sequenced, of which 15 were type G1 and 17 were type G5, with a length of 824 bp and 807 bp, respectively. After submitting to GenBank database, the login numbers obtained were OP413393-OP413402, OP413498-OP413506 and OP420520. A total of 34 nd1 genes were sequenced successfully, of which 11 were type G1 and 23 were type G5, with lengths of 882 bp and 888 bp, respectively. The entry numbers obtained by submitting the GenBank database were OP471626-OP471638. The results of sequence polymorphism analysis showed that 1.94% (16/824) mutation sites in the co1 type G1 sequence and 3.10% (25/807) mutation sites in the G5 type sequence were found. In the nd1, 0.45% (4/882) and 2.93% (26/888) mutation sites were found in the type G1 sequence and the G5 type sequence, respectively. The results of multiple genotypic sequence comparison showed that there were 6 homologous sequences of the co1 gene, 4 of which were type G1, the genetic distances of homologous sequences were 0.000 0-0.001 1, 0.000 0-0.000 6, and 0.000 0-0.001 7, respectively. Also included two G5 types. The genetic distances of homologous sequences were 0.000 0-0.002 5 and 0.000 0-0.001 2, respectively. There were 3 homologous sequences in nd1 gene, of which 1 was type G1, and the genetic distance was 0.000 0-0.001 1, 2 of which were type G5, genetic distance was 0.000 0 and 0.000 0-0.001 1. Phylogenetic tree analysis showed that the phylogenetic tree based on co1 and nd1 genes formed two branches of type G1 and type G5, type G1 was closely related to the gene sequences of central and western China (Qinghai, Ningxia, Gansu, Sichuan), Bhutan, Iran and Jordan in the Middle East (GenBank entry number AF297617.1, KJ628328.1, EU072106.1, MW138946.1, AB688602.1). The G5 Type was closely related to Vietnam, China's Guangxi, the United Kingdom, Poland, Zambia, France, Pakistan, Brazil, Kenya and Namibia (GenBank login numbers MW558412.1, MN058591.1, KU378107.1, MZ322608.1, KU743915.1, KU743919.1, MN886291.1, KX010903.1, KU743918.1 and KX138068.1). Conclusion The prevalent isolate of E. granulosus in Yunnan Province are of genotype G1 and G5, and both co1 and nd1 gene reveals single nucleotide polymorphisms.

Effect evaluation and factor analysis of ultrasonic manifestations in the diagnosis of hepatic alveolar echinococcosis

RAOWAN Tuolehong, ABUDUSALAMU Abulikemu, YANG Lingfei, CHEN Lu, LI Zhao, JIA Fang, SONG Tao

2023, 41(3):  312-318.  doi:10.12140/j.issn.1000-7423.2023.03.008

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Objective To understand the different ultrasonic manifestations of hepatic alveolar echinococcosis (HAE), to provide a reliable basis for improving the detection and diagnostic accuracy of HAE under conventional ultrasonography. Methods From January 2018 to April 2021, the patients who underwent routine ultrasound examination in the First Affiliated Hospital of Xinjiang Medical University and were diagnosed as HAE through surgery and pathology were recruited in the study. The relevant data, including gender, age, nationality, and ultrasonic image classification were collected. The clearest and complete images of HAE lesions in the original ultrasound images of the study subjects were selected for analysis, and the ultrasound manifestations of the lesions (location, size, boundary and shape, solid portion echo, calcification, liquefaction necrosis, blood flow signals, bile ducts and vascular invasion, etc.) were recorded, and values were assigned. Multiple lesions were described separately. The coincidence between post-operation pathological findings and the ultrasound diagnosis of the lesion was determined. Univariate analysis was conducted on the various ultrasound manifestations of HAE lesions, with statistically significant factors set as independent variables, while the coincidence with ultrasound diagnosis of lesions as dependent variables. A multivariate logistic regression analysis was conducted to establish a regression model, which was validated with the receiver operating characteristic (ROC) curve. The nomogram was constructed based on the results screened using multivariate logistic regression analysis, and its performance was evaluated using the ROC curve, calibration curve and decision analysis curve. Results The total number of HAE patients included in this study was 141. The age ranged from 9 to 65 years old, with an average age of (37.4 ± 13.6) years. There were 71 males and 70 females in this study, including 87 Tibetan patients accounting for 61.7%. Among the 141 HAE patients, 28 had single lesions, and 113 had multiple lesions. The final number of HAE lesions with inclusive criteria was 170. The results of univariate analysis showed that the difference in lesion location was not statistically significant (χ² = 1.952, P > 0.05). The differences in lesion size, boundary and shape, internal echo, calcification, liquefaction necrosis, blood flow signal, and bile ducts and vascular invasion were statistically significant (χ² = 39.026, 18.601, 15.743, 47.205, 34.151, 6.597, 21.766, all P < 0.05). The 4 factors including lesion size (OR = 0.180, 95% CI: 0.020-1.645), calcification (OR = 0.037, 95% CI: 0.002-0.590), liquefaction necrosis (OR = 0.282, 95% CI: 0.042-1.867), and blood flow signal (OR = 20.746, 95% CI: 3.720-115.686) were correlated with the ultrasound diagnosis compliance of HAE lesions. The accuracy analysis results of the logistic regression model predicted that the sensitivity of the regression model was 96.7% (118/122), the specificity was 83.3% (40/48), the positive predictive value was 93.7% (118/126) and the negative predictive value was 90.9% (40/44). The area under the ROC curve was 0.918 (95% CI: 0.859-0.977), the sensitivity was 95.1% and the specificity was 85.4%. The area under the ROC curve of the nomogram constructed based on the results of multivariate logistic regression analysis was 0.833. The sensitivity was 86.9% and the specificity was 70.8%. The calibration curve was close to the reference curve, and the decision analysis curve deviated significantly from both reference curves. Conclusion The size, calcification, liquefaction necrosis, and blood flow signal of the HAE lesions were important factors affecting ultrasound diagnosis.

Surveillance of echinococcosis in Qinghai Province in 2020

WANG Wei, CAI Huixia

2023, 41(3):  319-324.  doi:10.12140/j.issn.1000-7423.2023.03.009

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Objective To understand the epidemic situation of echinococcosis in Qinghai Province, to provide the scientific basis for optimization of prevention and control measures. Methods Echinococcosis surveillance was performed among residents, primary school students, intermediate hosts, definitive hosts, and questionnarire survey was conducted for primary school students in 39 echinococcosis endemic counties in Qinghai Province in 2020 (32 in typeⅠ and 7 in type Ⅱ). One or several nearby administrative villages (natural villages) were randomly sellected from each surveillance county as the surveiooance sites, each of which will not be repeated within 5 years. No less than 1 000 residents were surveyed at each surveillance, and examined using abdominal ultrasonography. From places near the surveillance site in the county, 1-5 primary schools were selected, in which all students in Grades 1 and 6 were examined by abdomen ultrasound scanning, with at least 500 students being examineds. A total of 300-500 cattle or sheep raised in the county were sampled from the surveillance county’s centralized slaughterhouse, and examined by inspection with naked eyes or palpation on the cystic matter or hard nodule of liver and the lung, with which autopsy was followed to determine Echinococcus infection. In each surveillance county with endemic alveolar echinococcosis, ten areas having people’s activity were selected to catch small rodents. From each selected area, no less than 50 animals were examined by dissection of livers and lungs to determine Echinococcus multilocularis infection. Ten dog owners were randomly selected from each administrative village in all epidemic townships in the surveiooance county to collect fresh domestic dog feces sample, one from each owner, while in type Ⅰ endemic county additional fecal samples of stray dogs and wild canines were collected. The Echinococcus antigen was detected using dog coproantigen detection kit. A questionnaire survey on echinococcosis prevention and control knowledge and behavior was undertaken in students of several classes selected from grade 3 to grade 6 in the schools under surveiooance scheme, with no fewer than 300 persons being surveyed in each county. Results In 2020, a total of 73 191 individuals in Qinghai Province were investigated by B-ultrasound, and 294 patients were with echinococcosis positive (including 5 new cases), with an overall prevalence of 0.40%. The prevalence in local residents and school children was 0.59% (276/46 922) and 0.07% (18/26 269), respectively. The residents and school children in type Ⅰ endemic counties had a prevalence rate of 0.70% (276/39 364) and 0.08% (18/22 646), respectively. There were no echinococcosis patients among type Ⅱ inhabitants or primary school students, showing a significant difference in the prevalence of echinococcosis between type Ⅰ and Ⅱ endemic counties (χ² = 53.32, P < 0.01). A total of 11 626 domestic animals were examined, and the total echinococcosis infection rate was 0.55% (64/11 626). The infection rate among sheep was 0.28% (18/6 491) and that among cattle was 0.89% (46/5 135). A total of 7 121 small rodents were investigated in the alveolar echinococcosis endemic county, and the infection rate of E. multilocularis was 0.38% (27/7 121). The positive rate of Echinococcus coproantigen was 0.54% (208/38 496) in 38 496 domestic dog feces samples. The positive rates of domestic dogs in type Ⅰ and Ⅱ endemic counties were 0.63% (177/27 882) and 0.29% (31/10 614), respectively, and the difference was not significant (χ² = 0.07, P > 0.05). A total of 7 253 fecal samples from stray dogs and wild canines were examined, and found coproantigen positive rate was 0.65% (47/7 253). Among them, the positive rates in type Ⅰ and Ⅱ endemic counties were 0.63% (43/6 853) and 1.00% (4/400) respectively, and the difference was not statistically significant (χ² = 0.82, P > 0.05). A total of 12 216 primary school students participated in the questionnaire survey. The awareness rate of knowledge on echinococcosis prevention was 97.8% (11 944/12 216), and the excellent rate was 88.6% (10 827/12 216). The awareness rates among students in type Ⅰ and Ⅱ endemic counties were 97.7% (9 865/10 096) and 98.1% (2 079/2 120) (χ² = 1.01, P > 0.05), and the excellent rates were 89.0% (8 986/10 096) and 86.8% (1 841/2 120) (χ² = 8.16, P < 0.01), respectively. The analysis of endemic situation of echinococcosis in different topographic areas of type Ⅰ endemic counties in Qinghai Province showed that the prevalence in people of Qingnan area, Qaidam Basin, arround the lake area and Hehuang Valley were 1.00% (261/25 980), 0.22% (21/9 354), 0.06% (10/17 200) and 0.02% (2/9 446). There was significant difference in the prevalence among residents in the four topographic areas (χ² = 271.31, P < 0.01). The positive rate of Echinococcus coproantigen from domestic dogs was 0.56% (34/6 043), 0.92% (21/2 283), 0.94% (62/6 613), and 0.46% (60/12 943) in the Qingnan area, Qaidam Basin, arround the lake area, and Hehuang Valley, respectively. The positive rate of coproantigen differed significantly between the area around the lake and Qingnan, Hehuang Valley, Qaidam Basin, and Hehuang Valley (χ² = 5.90, 15.86, 7.64; P < 0.05 or P < 0.01). The positive rates of coproantigen in stray dogs and wild canines were 0.75% (24/3 190), 0.38% (3/800), 0.72% (13/1 812) and 0 (0/1 051), respectively. There were significant differences in the positive rates of fecal antigen between street dogs and wild canines in Qingnan area, around the lake area and Hehuang Valley (χ² = 7.95, 7.58, P < 0.01). The infection rate of cattle in Qaidam Basin was the highest at 1.71% (12/700), which was significantly different from that in Qingnan area 0.87% (20/2 307) and around the lake area 0.57% (7/1 228) (χ² = 4.22, 6.58, P < 0.05). Alveolar echinococcosis endemic counties are mainly distributed in Qingnan area and around the lake area, where the infection rate of Echinococcus in small rodents was 0.39% (24/6 121) and 0.60% (3/500), respectively (χ² = 0.49, P > 0.05). Conclusion Both echinococcosis patients and new cases in Qinghai Province are distributed in typeⅠendemic counties in southern Qinghai Province, and the transmission cycle has not been entirely interrupted. The risk of infection in the population remains.

Investigation of fecal parasite pathogens in domestic dogs based on high-throughput sequencing

XIE Yi, WANG Ying, WANG Xu, SHI Dandan, FU Meihua, LI Chunyang, WU Weiping, DAN Bazeli, LIAO Sa, ZHANG Kaige, DENG Xueying, GUAN Yayi

2023, 41(3):  325-330.  doi:10.12140/j.issn.1000-7423.2023.03.010

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Objective To investigate the parasitic infection in domestic dogs in Da Hinggan Ling area, and provide reference for the prevention and control of local zoonotic parasitic diseases. Methods In September 2020, fresh domestic dog feces were collected from dog owner’s household yard of 6 natural villages including Jiabei Village, Baihua Village, Dongshan Village, Tuqiang Town, Xingfu Village and Guyuan Town in the Da Hinggan Ling area, and total DNA was extracted from the fecal samples. PCR amplification was conducted using 13 pairs of parasite universal primers targeting 8 taxonomy groups including Apicomplexa, Amoeba, Diplomonadida, Kinetoplastida, Parabasalia, Nematoda, Platyhelminthes and Microsporidia, and the target gene fragments were obtained after high-throughput sequencing of the amplified products. The product sequences were compared and analyzed in the NCBI database to identify the parasite species, and the detection rate of parasites in domestic dog feces was calculated. Results Parasite DNA were detected in 37 of 202 domestic dog feces, and the total detection rate was 18.31%. A total of 19 species of parasites which belongs to 15 families from 7 phyla were detected, and the detection rate of different species were significantly different (χ² = 69.488, P < 0.05). Thirteen protozoa species and 6 helminth species were detected. The detection rate of Parabodo caudatus in the family Parabodonidae was the highest at 6.44% (13/202). Among the 37 samples in which parasite DNA was detected, the mixed positive ratio was 32.43% (12/37), of which 72.97% (27/37) detected 1 type of parasite, and 2 types, 3 kinds and 4 kinds accounted for 27.03% (10/37), 0.27% (1/37) and 0.27% (1/37) respectively. Jiabei Village had the highest detection rate of parasite DNA in domestic dog feces, which was 37.70% (23/61), followed by Tuqiang Town, with a detection rate of 36.36% (20/55), and Baihua Village, which was not detected (0/13). There was a statistically significant difference in the detection rate of different sampling sites (χ² = 19.717, P < 0.05). Conclusion In Da Hinggan Ling area, domestic dogs were found infected with multiple parasites, showing considerably higher positive rate detected in some parts of the area, thus, there exists risk of infection caninee-source parasites to local residents.

Epidemiological analysis of soil-transmitted nematode infections in China in 2020

ZHANG Mizhen, HUANG Jilei, ZHU Huihui, ZHOU Changhai, ZHU Tingjun, QIAN Menbao, CHEN Yingdan, LI Shizhu

2023, 41(3):  331-335.  doi:10.12140/j.issn.1000-7423.2023.03.011

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Objective To understand the situation of soil-transmitted nematode infection in China in 2020 and provide support for evaluating the development of surveillance on soil-transmitted nematodiasis in various provinces/municipalities/autonomous regions, and improving and perfecting the control strategies. Methods Surveillance was carried out in 408 national surveillance sites (counties) in 31 provinces (municipalities, autonomous regions) in China in 2020. With the county as unit, each site was divided into 5 areas geographically: east, west, south, north, and central part, followed by selecting one township (town), and therein one administrative village was selected from wherein, 200 permanent residents over the age of 3 were sampled. A total of 1 000 people were surveyed at each surveillance site. Fecal samples were collected from the sampled villagers, and examined by using the modified Kato-Katz thick smear method (two slide-reading for each sample) for infection of hookworm, Ascaris lumbricoides, Trichuris trichiura and Enterobius vermicularis, to calculate the infection rate and intensity, respectively. In addition, soil samples were collected from fields or vegetable gardens of each village in the survey site, and examined for hookworm larvae using 5% saline at 45 ℃, and for Ascaris eggs by saturated sodium nitrate flotation method. Results In 2020, the overall infection rate of soil-transmitted nematode in residents was 0.84% (3 485/415 672) in 408 surveillance sites of 31 provinces (municipalities/autonomous regions), with the highest found in Hainan (6.34%, 199/3 141), followed by Yunnan (5.80%, 963/16 616) and Sichuan (3.66%, 592/16 168); infection rate in females was 0.91% (1 944/213 591), which was higher than that of 0.76% in males (1 541/202 081) (χ² = 27.20, P < 0.01). The soil-transmitted nematode infection rate was the highest in the age group ≥ 60-years-old, which is 1.26% (1 376/109 251). The difference between each age group was statistically significant (χ² = 382.28, P < 0.01). The infection rates of hookworm, A. lumbricoides, T. trichiura were 0.51% (2 016/415 672), 0.19% (805/415 672) and 0.16% (673/415 672), respectively. Among them, hookworm and T. trichiura had only mildly infected cases. The proportions of mild and moderate A. lumbricoides infections were 99.25% (799/805) and 0.75% (6/805), respectively. In 2020, 2 604 soil samples were examined and found that the positive rate of Ascaris eggs and hookworms was 3.07% (80/2 604) and 2.42% (63/2 604), respectively. Conclusion In 2020, the infection rate of soil-transmitted nematode in China remains at a low level in general, but the regional differences are still significant, and the areas with high infection rates still exist. At the same time, it is necessary to strengthen the control measures for the key groups of people over age of 60, women and children, and carry out health education.

Effects of soil-transmitted nematode infection on wheezing, asthma, and skin prick test positivities in rural preschool children

LI Fang, WANG Shufang, HE Pengfei, XU Shuhui

2023, 41(3):  336-343.  doi:10.12140/j.issn.1000-7423.2023.03.012

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Objective To investigate the effects of soil-transmitted nematode infection of mother and child on wheezing, asthma, and skin prick test (SPT) positive in rural preschool children. Methods Preschool children and their mohers in rural area were enrolled respectively, who underwent physical examination in the Yuncheng Central Hospital outpatient sector from April 2020 to March 2021. Basic information of mothers and children and children’s wheezing/asthma condition were collected through questionnaires. Fecal samples were collected from participants, and examined for soil-transmitted nematode eggs by the Kato-Katz thick smaear method, The SPT test was performed for children to collect allergic and atopic data. Multivariate unconditional logistic regression analysis was used to explore the relevant factors affecting asthma, wheezing, and SPT positivity in children. Results In this study, there were 2 014 children and their mothers each. The infection rate of soil-transmitted nematodes among the mothers was 14.40% (290/2 014), and among the preschool children was 7.99% (161/2 014). There were 117 children had wheezing symptoms, with an occurrence rate of 5.81% (117/2 014); 149 cases had asthma, with an occurrence rate of 7.40% (149/2 014) and 304 cases were positive for SPT, with a positive rate of 15.09% (304/2 014). Premature infants, mothers with a history of allergies, and family members smoking are risk factors for wheezing in children (P < 0.05, OR > 1); female gender and complete breastfeeding are protective factors for wheezing in children (P < 0.05, OR < 1). Mothers with a history of allergies, having someone smoking at home, and planting flowers and plants at home are risk factors for childhood asthma (P < 0.05, OR > 1); female and complete breastfeeding are protective factors for childhood asthma (P < 0.05, OR < 1). Mothers with a history of allergies is a risk factor for SPT positivity in children (P < 0.05, OR > 1), while the gender of the child, presence of soil-transmitted nematode infection in the child, and presence of soil-transmitted nematode infection in the mother are protective factors for SPT positivity in children (P < 0.05, OR < 1). Conclusion Mothers and children infected with soil-transmitted nematode may reduce the occurrence of allergic diseases in children.

Morphological observation and phylogenetic analysis for adult Histiostoma feroniarum

TANG Feifan, JIANG Feng, WANG Meiqing, WANG Lan, CAO Jiacheng, XIA Yuting, WANG Jingxuan, TANG Liqing, ZHAN Xiaodong

2023, 41(3):  344-348.  doi:10.12140/j.issn.1000-7423.2023.03.013

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Objective To observe the morphological characteristics of adult Histiostoma feroniarum under the electron microscope, and to study the genetic relationship between this species and other mite species of Acaridae. Methods The adult stage of H. feroniarum was isolated from the culture, and its external morphological features were observed using scanning electron microscopy (SEM), followed by extracting the mite DNA, for amplifying the cytochrome oxidase subunit 1 (cox1) and internal transcribed spacer (ITS) by PCR. The ITS and cox1 gene was sequenced and compared with the other 8 Acaridae species in GenBank. The phylogenetic tree was constructed by the maximum likelihood method. Results Under the electron microscope, the ventral epidermal protrusions of the mites were more developed. The epidermal protrusions of foot I healed into the chest plate, and the epidermal protrusions of foot Ⅱ extended to the centre and were not connected. There were 2 pairs of chitinous rings on the ventral surface, and the male adults were located between the Ⅱ-Ⅳ basal ganglia. The first pair of chitin rings of the female mite was located between the foot Ⅱ and Ⅲ, and the last pair of chitin rings were located at the level of the foot Ⅳ basal ganglia. Compared with the light microscope, the structures of male reproductive folds, leaflets and penis were clearer under the electron microscope. The cox1 and ITS sequences were obtained by PCR amplification, which was 539 bp and 1 633 bp. The ITS sequence similarity comparison results showed that the sequence similarity between H. feroniarum and Blomia tropicalis (GenBank accession no. KC215362) was the highest, which was at 90.41%, and the sequence similarity with other Acaridae species was 83.33%-89.73%. The cox1 sequence alignment showed that the highest sequence similarity was 83.55% between H. feroniarum and Lepidoglyphus destructor (GenBank accession no. MT075728), and the sequence similarity with other Acaridae species ranged from 79.78%-82.12%. The phylogenetic tree constructed based on ITS and cox1 sequences has differences in topological structure. showing that H. feroniarum is a single branch, but not at the same branch of other Acaridae species, which is likely consistent with morphological classification. The phylogenetic analysis based on ITS showed that H. feroniarum and Rhizoglyphus robini were clustered as a branch, which differs from morphological classification. Conclusion The morphological structure of male and female H. feroniarum could be clearly observed by scanning electron microscopy. Phylogenetic analysis of cox1 sequence reveals relatively distant genetic relationship seen between the H. feroniarum and other acaroid mites.

TEACHING RESEARCH

Exploration of the co-teaching mode of basic-clinical collaboration in medical parasitology class

SHEN Yan, WANG Jun, LIANG Jiao, ZHAO Ya, LI Yinghui

2023, 41(3):  349-354.  doi:10.12140/j.issn.1000-7423.2023.03.014

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There are limitations in the traditional medical teaching model, which hinder students from integrating basic knowledge and clinical skills, and also make the teaching of basic science and the teaching of clinical practise to split for a long time. Medical science teachers and clinical medical teachers lack communication and knowledge complementarity. Through the initial attempt and exploration of the dual-teacher teaching model in the medical parasitology course, the whole process from the content selection and case selection to classroom design has been formed. While summarizing the teaching effect, some problems that need to be improved have also been exposed. Double runs in teaching mode to build diversified class in the main body, increasing the interaction between the main body, through the “teacher-teacher” dialogue triggered the “teacher-student” interaction and then the “student-student” interaction, create good teaching situation, help students to cultivate diverse thinking, promote the multidirectional flows of knowledge, compensate for the limitations of the basic medical education, enrich students’ knowledge and clinical thinking infiltration on the early clinical diagnosis and treatment. The teaching mode of dual teachers puts forward higher requirements on the tacit understanding of teachers, forcing basic science teachers and clinical teachers to improve their teaching ability through full communication and mutual learning, re-combing classroom design, realizing the combination of basic science knowledge and clinical knowledge, and realizing teaching and learning. At the same time, the dual-teacher classroom teaching mode puts forward higher requirements on the clinical knowledge reserve and the learning ability before, during and after class, and requires the students to develop the habit of thinking, which is conducive to the cultivation of clinical thinking.

REVIEWS

Effect of tourism development on schistosomiasis control in China

LI Jie, WEN Yusong, LI Zhaojun

2023, 41(3):  355-360.  doi:10.12140/j.issn.1000-7423.2023.03.015

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Currently, the schistosomiasis control program is moving from transmission-control and transmission-blocking towards elimination in China. However, the rapid development of countryside tourism will inevitably pose impacts on the consolidation of schistosomiasis control achievements and elimination campaigns. This review summarized the schistosomiasis epidemiology and tourism development in schistosomiasis-endemic areas in China, as well as the impact of tourism development on schistosomiasis control in endemic areas. We have proposed new intervention models for schistosomiasis control and elimination in endemic areas and ecological schistosomiasis control policies that adapt to rural tourism development.

Research progress in adjunctive therapy of cerebral malaria

CAO Wei, WANG Yi, ZHANG Xizhi, TONG Guodong, YANG Chao, SHEN Yan, ZHAO Ya

2023, 41(3):  361-373.  doi:10.12140/j.issn.1000-7423.2023.03.016

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Cerebral malaria is one of the major fatal complications of Plasmodium falciparum infection, and the pathogenesis is not fully understood. However, most of the current studies believe that the main pathogenesis of cerebral malaria is the obstruction of cerebral microcirculation caused by infected red blood cells adhering to cerebrovascular endothelial cells and the resulting immunopathological damage caused by the activation of cerebrovascular endothelial cells and the infiltration of inflammatory cells, which synergistically result in the disruption of blood-brain barrier and brain edema. Therefore, the magnitude of immune activation and damage to the cerebrovascular endothelial cells is one of the key factors in determining the progression and outcome of cerebral malaria. Existing clinical studies have proved that even active and effective anti-malarial treatment with artemisinin-based drugs cannot completely alleviate the celebral malaria symptoms and the possible subsequent neurological tissue damage, while the combination with anti-inflammatory treatment of glucocorticoids may cause serious side effects. The study on combination therapies of cerebral malaria involves multiple links for relevant pathogenesis, while the existing adjunctive therapies have not achieved the expected curative effect due to various factors, such as the complex life cycle and high antigenic variation of P. falciparum. Therefore, new combination therapies are urgently needed. This paper provides a brief review of the current progress in combination therapies against cerebral malaria with the target of protecting cerebrovascular endothelial cells by enhancing the self-protection function of endothelial cells and inhibiting the damage of other inflammatory cells or harmful factors to endothelial cells.

Research progress on transcription regulation of rif gene in Plasmodium falciparum

XU Shaojie, CHEN Shenbo, CHEN Junhu

2023, 41(3):  374-379.  doi:10.12140/j.issn.1000-7423.2023.03.017

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Plasmodium falciparum-infected erythrocytes express multiple polymorphic proteins known as variant surface antigens, which are associated with symptomatic and severe malaria. The repetitive interspersed family (rif) gene is one of the largest gene families of the P. falciparum multicopy gene families which potentially encode variant antigens. Studying the regulation of rif genes, such as histone modification, repressor element regulation and the involvement of expressed proteins in immune evasion, is of great significance. Based on the current research, we made a comprehensive summary of the rif gene expression regulation mechanisms to understand the pathogenic mechanism of P. falciparum.

SHORT COMMUNICATIONS

Molecular identification and genetic tracing of Giardia lamblia isolated from an infected case

WANG Dan, HE Zhiquan, LIU Ying, LIU Lingzhi, CHEN Huihui, JIANG Tiantian, JI Penghui, QIAN Dan, YANG Chengyun, ZHANG Hongwei

2023, 41(3):  380-383.  doi:10.12140/j.issn.1000-7423.2023.03.018

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To investigate the molecular identification of Giardia lamblia isolated from an infected case and analyze the source of infection. Fresh fecal samples of the patient and his parents and the nanny were collected for iodine solution staining and microscopic examination. Fecal DNA was extracted, and nested PCR was used to amplify the triosephosphate isomerase (tpi), glutamate dehydrogenase (gdh) and beta-giardin (bg) gene for sequencing. The homology comparison and phylogenetic analysis of gene sequences were performed by BLAST, ChromasPro and MEGA 11.0 software to determine the assemblages. The results showed that there were trophozoites and cysts in the fecal of the patient through microscopic examination. Nested PCR amplified bands of about 500 bp, which were consistent with the tpi, gdh and bg gene fragments of G. lamblia, confirming the infection of G. lamblia. None of the patient’s parents and nanny had diarrhea symptoms, but G. lamblia cysts were found in the stool specimen of his father. Combined with nested PCR and sequencing results, the father was a G. lamblia carrier. The results of the gene comparison showed that the percent identity of tpi, gdh and bg genes sequence amplified from fecal samples of the patient and his father was 99.8%, 100% and 98.5%, respectively. The percent identity of tpi gene sequence amplified from the fecal samples of the patient and his father showed 100% and 99.8% with G. lamblia assemblages AⅡ (GenBank accession no. LC183963), respectively. The amplified gdh gene sequence showed 99.6% and 100% with assemblages AⅡ (GenBank accession no. KF843931), respectively. The percent identity between the bg gene sequence amplified from the patient fecal sample and the sequence of assemblages AⅢ (GenBank accession no. LC183968) is 99.2%, while the percent identity between the bg gene sequence amplified from the father’s fecal sample and the sequence of assemblages AⅡ (GenBank accession no. LC183975) is 100%. The results of phylogenetic tree analysis showed that both the patient and his father were infected with G. lamblia and clustered in the same cluster as assemblages A, and sub-genotype analysis using tpi and gdh as target genes were conducted, which clustered in the same cluster as assemblages AⅡ. Therefore, it can be determined that the patient was infected with G. lamblia assemblages AⅡ, and it could be a family cluster infection caused by internal transmission within the family.

Epidemiological analysis of malaria in Guizhou Province from 2017 to 2021

GENG Yan, LAN Ziyao, LI Yang, DAI Jiarui, CAI Shan, LU Lidan, HUANG Yuting, SHI Weifang, SHE Danya

2023, 41(3):  384-388.  doi:10.12140/j.issn.1000-7423.2023.03.019

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To understand the malaria epidemiological characteristics in Guizhou Province from 2017 to 2021, the malaria endemic data and individual case information in Guizhou from 2017 to 2021 were collected and analyzed using descriptive statistical methods, species of Plasmodium, source of infection, distribution of cases, clinical visits and diagnosis were analyzed. According to the analysis, 84 malaria cases were reported in Guizhou from 2017 to 2021. Most cases were P. falciparum infection (69.0%, 58/84). All the cases were imported from overseas, 91.7% (77/84) of which were imported from African countries and others were imported from Asia and Oceania. The cases were reported throughout the year, particularly higher in January (15.5%, 13/84). All the cities in Guizhou Province had malaria cases reported, of which Guiyang had the highest number of reported cases (41.7%, 35/84). Of all the reported cases, 92.9% (78/84) were male, and 69.0% (58/84) were aged 30 to 49; 73.8% of the cases were initially diagnosed as malaria. The Center for Disease Control and Prevention (CDC) at the county level had the highest diagnostic accuracy (15/15). The median interval from the onset of symptoms to initial diagnosis was 1 day, and from initial diagnosis to final diagnosis was 3 days. All the cases were reported timely within 1 day after diagnosis, 90.5% (76/84) were undertaken epidemiological investigation within 3 days after reporting, then 98.8% (83/84) of the epidemic sites were surveyed and performed prevention measures within 7 days. This study suggested that to consolidate the achievement of malaria elimination in Guizhou, continuous surveillance of imported malaria cases and regulated disposal measures should be maintained, and the diagnostic and tertiary care capacity should be improved.

Seroepidemiological survey of Toxoplasma gondii infection in dogs and cats in Beijing 2022

DU Juan, LI Jia, WU Di, YU Qi, ZHANG Wei, BAI Runian, GUO Junlin, LIU Qingbin, LEI Qili, GU Chuanhui, WANG Meng, ZHAO Haojun

2023, 41(3):  389-392.  doi:10.12140/j.issn.1000-7423.2023.03.020

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In order to identify the infection of Toxoplasma gondii in dogs and cats in Beijing, according to the program of animal disease surveillance in Beijing in 2022, 920 dog serum samples and 816 cat serum samples were collected from scatters, animal hospitals and farms in the whole city. Indirect heamagglutination assay were uesd to detecte Toxoplasma gondii antibodies in dog and cat serum samples. The result shows that the T. gondii antibody positive rate was 5.2% (48/920) in dogs and 7.5% (61/816) in cats. The positive rate of T. gondii antibody in dogs in autumn was 6.8% (30/440), which was higher than that in spring at 3.8% (18/480) with moderate intensity correlation (χ² = 4.37，P < 0.05，0.4 < OR < 0.6). And the positive rate of T. gondii antibody in cats in autumn was 12.5% (50/401), which was higher than that in spring at 2.7% (11/415) with strongly correlated material (χ² = 28.42，P < 0.05，0.1 < OR < 0.3）. In diverse sources, the positive rates of T. gondii antibody in dogs and cats from veterinary clinics were the highest, which were 8.0% (47/590) and 8.5% (61/715) (χ² = 25.31, 9.31，all P < 0.05), respectively. The positive rate of T. gondii antibody in dogs from the six central districts was 11.4%（40/350）. The positive rate of T. gondii antibody in dogs from the outer suburban districts was 1.4% (8/570) with strongly correlated material (χ² = 44.07，P < 0.05，3.0 < OR < 9.9). And the positive rate of T. gondii antibody in cats from the six central districts was 10.3% (37/360). The positive rate of T. gondii antibody in cats from the outer suburban districts was 5.3% (24/456) with moderate intensity correlation (χ² = 7.31，P < 0.05，1.5 < OR < 2.9). This study provided a basis for the further prevention and control of toxoplasmosis in Beijing.

CASE REPORTS

A case of late stage tumor combined with Babesia infection

XU Yile, LI Jiangfeng, ZHANG Jiaqi, YU Xianghua, RUAN Wei

2023, 41(3):  393-396.  doi:10.12140/j.issn.1000-7423.2023.03.021

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A 57-year-old female patient from Lucheng District was admitted to Wenzhou Central Hospital on September 28, 2021. The examination on admission showed the chronic disease face and clear consciousness; the skin and mucous membranes had no yellowish staining nor oedema but were scared. The patient was admitted to the cardiology department with “chest tightness pending further investigation” and was later transferred to the chemotherapy department due to cancerous pleural fluid. The patient had a recurrent fever since October 8. Laboratory tests showed the haemoglobin was 86 g/L, the red blood cell count was 2.74 × 10¹²/L suggesting moderate anaemia. The leukocyte count was 12.7 × 10⁹/L, the absolute neutrophil count was 10.6 × 10⁹/L. The patient also had mildly elevated total and indirect bilirubin; urine culture detects mixed growth of more than 3 bacteria, persistent urinary occult blood 2 +. Wright’s stain of the blood smear was performed on October 25, microscopic examination showed that 1-4 ringlets with purplish-red nuclei and blue cytoplasmic could be found in some red blood cells, which was considered to be a parasitic infection. The patient was a long-term resident of urban Wenzhou with no history of domestic or international travel, and no clear history of tick bites. The patient underwent breast-conserving surgery for left breast cancer in 2005, received chemotherapy and targeted drug therapy for multiple lung metastases between 2014 and 2021 and had 12 blood transfusions from June to October 2021. The PCR product of blood DNA amplificated by primers specific to Babesia was sequenced and identified with 99.43% homology to B. microti (GenBank accession: MG674832.1). The diagnosis of Babesia microti infection was confirmed. The patient was treated with oral chloroquine phosphate tablets (0.5 g/d, double the first dose for 5 d) and intravenous clindamycin (600 mg/d + saline 500 ml intravenously for 10 d), with some relief of symptoms, and parasites were still detected on microscopic examination after 5 days. The symptoms of high fever, jaundice and haemolysis caused by Babesia infection somewhat exacerbate the progression of terminal-stage cancer. On November 8, the patient died of multi-organ failure and unsuccessful resuscitation.