Current Issue
30 December 2023
Volume 41 Issue 6
CONSENSUS AND GUIDELINE
Expert consensus on diagnosis and treatment of food-borne parasitic diseases (2023)
Expert Group of National Center for Infectious Diseases, National Center for Infectious Disease Medicine
2023, 41(6):  653-668.  doi:10.12140/j.issn.1000-7423.2023.06.001
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Food-borne parasitic diseases caused by ingesting food and water containing infective parasites are still common parasitic diseases that are easily misdiagnosed and mistreated in clinical practice. With the participation of multi-disciplinary experts, and in the light of the latest research results at home and abroad, based on factors other than the quality of evidence (economics, patient preferences and values, trade-offs, accessibility, fairness, acceptability, etc.), the level of recommendation and the quality of evidence in evidence-based medicine were assessed using the World Health Organization-recommended evidence quality classification and strength of recommendation system, and a consensus of 24 items was reached to guide and improve the comprehensive diagnosis and treatment of food-borne parasitic diseases for clinical medical staff.

ORIGINAL ARTICLES
Epidemiological characteristics of visceral leishmaniasis in China in 2022
LI Yuanyuan, ZHOU Zhengbin, YANG Limin, LI Zhongqiu, LIU Qin, ZHANG Yi, LI Shizhu
2023, 41(6):  669-676.  doi:10.12140/j.issn.1000-7423.2023.06.002
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Objective To understand the epidemic situation of visceral leishmaniasis in China in 2022 and provide scientific basis for formulating prevention and control strategy. Methods Data of visceral leishmaniasis in 2022 was collected from the web-based National Infectious Diseases Reporting Information Management System. After excluding suspected cases, duplicates and cutaneous leishmaniasis cases, a visceral leishmaniasis database was established and underwent descriptive epidemiological analysis with Microsoft Excel 2016. Results A total of 239 cases of visceral leishmaniasis were reported in 104 counties of 11 provinces (autonomous regions and municipalities) in 2022, among them 191 cases were reported from mountain-type zoonotic visceral leishmaniasis endemic areas, 4 cases from desert-type zoonotic visceral leishmaniasis endemic areas, 2 cases from anthroponotic visceral leishmaniasis endemic areas and 42 cases were imported from non-endemic areas. These cases were mainly distributed in Shanxi (110 cases), Shaanxi (34 cases), Henan (23 cases) and Hebei (23 cases), the total reported accounting for 79.30% (190/239) of the overall total in China. A total of 197 local transmitted cases were reported from 42 endemic counties and other 63 counties were of non-endemic areas, reporting 39 imported cases. Pingding County, outer suburbs of Yangquan City, Jingxing County and Huazhou District were the major endemic counties with 25, 15, 16 and 10 cases reported respectively, the total accounting for 27.62% (66/239) of the overall total cases in China. Recurrence endemic counties of visceral leishmaniasis were mainly concentrated in Shanxi (Qinshui County, Gaoping City, Fushan County, Houma City, Yicheng County, Jiangxian County, Yuanqu County, Wenshui County), Hebei (Mining district of Jingxing County, Zanhuang County, Lincheng County, Xindou District), Henan (Yanshi District, Shangjie District, Qibin District), Beijing (Changping District) and Xinjiang (2th regiment farm) in 2022, with a total of 23 local cases reported. The peak incidence occurred in July. The ratio of male to female cases was 1 ∶ 0.38. Farmers, infants and young children are the high-risk population of visceral leishmaniasis, accounted for 53.97% (129/239) and 16.74% (40/239) of the total cases respectively. The reported cases at age of ≥ 15 accounted for 81.17% (194/239). Conclusion Visceral leishmaniasis is at a low prevalence status in China, whereas the endemic area is gradually expanding, and the number of cases is gradually increasing in mountain-type zoonotic visceral leishmaniasis endemic area.

Identification and analysis of host proteins in the cyst wall and cyst fluid of Echinococcus granulosus in human infection
LI Tao, LI Zihua, ZHANG Cuiying, ZHAO Wei
2023, 41(6):  677-682.  doi:10.12140/j.issn.1000-7423.2023.06.003
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Objective To further elucidate the interaction between Echinococcus granulosus and the host by identifying the host protein components in the cyst wall and cyst fluid of E. granulosus. Methods The active and intact echinococcal cysts removed surgically from 10 patients with E. granulosus were collected from the Department of Hepatobiliary Surgery, Ningxia Medical University General Hospital in 2022. Blood samples were also taken from the same patients. The transparent cyst fluid in the ascus was extracted aseptically, and centrifuged to collect the supernatant; the inner wall of the ascus was repeatedly washed, cut into pieces, lysed and homogenised to collect the supernatant. The bicinchoninic acid (BCA) protein quantification kit was used to extract the cyst fluid and cyst wall total protein. The cyst wall total protein lane was cut out after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) electrophoresis. The significant protein bands were analysed by mass spectrometry and Western blotting analysis. Quantitative analysis was performed to determine the biochemical component concentrations in the patient serum and cyst fluid. Results The liver function of 10 patients with echinococcosis granulosus was Child-Pugh A or B. The diameter of the lesion ranged from 2.9 to 16.3 cm, which were classified as CE2 (multiple subcysts) or CE3 (collapsed internal capsule) according to the WHO-IWGE classification. SDS-PAGE electrophoresis analysis results suggested that the echinococcal cyst wall protein showed clear protein bands at the relative molecular mass (Mr) of 70 000, 50 000, and 25 000, and the cyst fluid protein showed clear bands at Mr 70 000. Protein bands. The protein spectrum and Western blotting results showed that the protein with a relative molecular mass of approximately Mr 70 000 contained human albumin, the target protein with a relative molecular mass of approximately Mr 50 000 was a human IgG heavy chain, and the target protein with a relative molecular mass of approximately Mr 25 000 was a human IgG light chain. Among the various biochemical indicators, the concentrations of K+ in cyst fluid and serum were (5.53 ± 0.86) mmol/L and (3.92 ± 0.33) mmol/L respectively. The albumin concentrations were (0.57 ± 0.46) g/L and (39.42 ± 2.77) g/L. The cholesterol concentrations were (0.03 ± 0.02) mmol/L and (3.79 ± 1.53) mmol/L respectively. The concentration differences between the three in cyst fluid and serum were statistically significant (t = 5.56, 43.71, 7.74; P < 0.05). Conclusion Host-derived serum albumin and antibodies are found of comparatively higher amont in the cyst wall and cyst fluid of E. granulosus.

Identification and analysis of miRNA targeting CYP450s genes in mosquitoes
WU Jiahui, SONG Xiao, CHENG Peng, LIU Hongmei, GUO Xiuxia, WANG Haifang, GONG Maoqing
2023, 41(6):  683-690.  doi:10.12140/j.issn.1000-7423.2023.06.004
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Objective To screen and identify the microRNAs (miRNAs) targeting cytochrome P450 enzymes (CYP450s) genes in mosquitoes, and analyze the differentially expressed miRNAs and the function of their target genes. Methods Ten larvae of each cypermethrin-susceptible (CS) and cypermethrin-resistant (CR) line Culex pipiens pallens mosquitoes and 5 blood-unfed female adult mosquitoes of each line on 3 d post-emergence were collected for extracting total RNA followed by sequencing, to calculate the differential expression between CS and CR lines using DEGseq software. MiRanda and RNAhybrid databases were used to screen the target genes of differentially expressed miRNAs, and gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis was performed. Five differentially expressed miRNAs (miR-11-5p, miR-317_3, miR-278-3p_1, miR-8-5p and miR-305-3p_5) that may regulate the expression of CYP450s genes were selected for real-time quantitative PCR (qRT-PCR) to validate the sequence output. The relative expression level of differentially expressed miRNAs target genes (CYP6a8, CYP6BB1v2, CYP6N22, CYP6N26P, CYP9b2) were detected by qRT-PCR. Results A total of 196 miRNAs were differentially expressed between CS and CR line Culex pipiens pallens. Among them, 16.3% (32/196) were differentially expressed in age Ⅲ larvae, 20.4% (40/196) in age Ⅳ larvae and 26.0% (51/196) in female adult mosquitoes exclusively; 12.2% (24/196) were differentially expressed in all stages. 45 were up-regulated and 46 were down-regulated of 91 miRNAs differentially expressed at age Ⅲ larvae. 59 were up-regulated and 45 were down-regulated of 104 miRNAs differentially expressed at age Ⅳ larvae. 44 were up-regulated and 54 were down-regulated of 98 miRNAs differentially expressed at female adult mosquitoes. The results of the GO and KEGG enrichment analysis showed that the target genes of differentially expressed miRNAs were mainly related to cellular, metabolic and single-cell biological pathways. The molecular functions of target genes were mainly involve with binding, catalytic activity and transporter activity. The target genes were mainly enriched in the pathways related to signal transduction, infectious diseases, cancers and parasitic diseases. The qRT-PCR results showed that the trends of 5 differentially expressed miRNAs were basically consistent with the sequencing results; The expression profiles of 5 miRNAs were opposite to the target genes (CYP6a8, CYP6BB1v2, CYP6N22, CYP6N26P and CYP9b2). Conclusion The five differentially expressed miRNAs screened in this study (miR-11-5p, miR-317_3, miR-278-3p_1, miR-8-5p and miR-305-3p_5) could regulate the insecticide resistance of Culex pipiens pallens by modulating the expression of CYP450s.

Drug-regulation of PPAR-γ/RXR-α signal pathways on ameliorating lung injury induced by Paragonimus proliferus infection in rats
HUA Lijuan, LI Shenghao, CHANG Guoji, LIU Siqi, DING Jie, BAI Baoli, ZHANG Lu, WANG Qingqing
2023, 41(6):  691-698.  doi:10.12140/j.issn.1000-7423.2023.06.005
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Objective To explore the effect of drug-regulation on peroxisome proliferator-activated receptor-γ/retinoid X receptor-α (PPAR-γ/RXR-α) signalling pathway to improve lung injury caused by Paragonimus proliferus infection in rats. Methods The P. proliferus were isolated from crabs collected in Yunnan Province, and SD rats were infected with 8 metacercariae per rat via subcutaneous injection at abdominal wall. The infected rats were divided into the high-fat-diet group, rosiglitazone group and bexarotene group, with 10 rats in each group, and were given with high-fat-feeding (high-fat diet), rosiglitazone [3 mg/(kg·d)], bexarotene [10 mg/(kg·d)] by gavage for 7 days. The infected and healthy rats were used as the infection control group and healthy control group. On 28 d post-mediated, the rats were sacrificed to collect lung tissue and heart apical blood. The lung tissue sections were prepared and stained with hematoxylin-eosin to assess the tissue damage and score the alveolitis semi-quantitatively. Serum interleukin-1β (IL-1β) and tumor necrosis factor α (TNF-α) levels were detected by ELISA. The lung tissue total protein was extracted for examining relative expression levels of PPAR-γ/RXR-α signalling pathway-related key target proteins indluding PPAR-γ, RXR-α, fatty acid transporter 3 (FATP3), apolipoprotein A1 (ApoA1), nuclear factor kappa-B (NF-κB) signalling pathway proteins [p65, activator protein (AP1)], janus kinase (JAK)/signal transducer and activator of transcription (STAT) signalling pathway proteins (JAK2, STAT3) by Western blotting. The inter-group data were compared using single-factor ANOVA analysis. Results All rats were successfully infected except 2 rats in the bexarotin group. Parasitic cysts were found in the lungs or the chest cavity. HE staining showed that in the infection control group, a large number of inflammatory cells infiltrated the alveolar septum, the alveolar wall thickened, and the alveolar cavity collapsed or closed. In the high-fat diet group, obvious inflammation injury was shown. Compared with the infection control group, the alveolar cavity inflation was slightly improved. The inflammatory cells of alveolar septum in the rosiglitazone group and bexarotene group were significantly reduced. Compared to the infection control group, the degree of alveolar wall thickening was significantly reduced, and the alveolar cavity inflation was significantly improved. The semi-quantitative alveolitis scores were 0.300 ± 0.053, 2.200 ± 0.189, 1.900 ± 0.320, 1.300 ± 0.301 and 1.500 ± 0.112 (F = 12.033, P < 0.05) in the healthy control group, the infection control group, the high-fat diet group, the rosiglitazone group and the bexarotene group, respectively. ELISA tests show that serum IL-1β levels in the healthy control group, infected control group, high-fat diet group, rosiglitazone group and bexarotene group were (103.23 ± 3.37), (111.59 ± 20.49), (110.13 ± 12.95), (89.91 ± 14.84) and (96.34 ± 19.03) pg/ml, respectively. TNF-α levels were (144.81 ± 1.35), (180.21 ± 23.38), (171.76 ± 27.83), (155.37 ± 13.67) and (143.24 ± 23.66) pg/ml, respectively. There was a significant difference among the five groups (F = 17.236, 13.558; P < 0.05). Western blotting showed that in the lung tissue of rats in the healthy control group, the infection control group, high-fatdiet group, rosiglitazone group, and bexarotene group, PPAR-γ relative expression level were 0.51 ± 0.09, 0.67 ± 0.06, 0.75 ± 0.08, 0.34 ± 0.02 and 0.56 ± 0.04, respectively; RXR-α were 0.89 ± 0.05, 0.15 ± 0.03, 0.81 ± 0.09, 0.22 ± 0.02 and 0.61 ± 0.10, respectively; FATP3 were 0.59 ± 0.06, 0.64 ± 0.06, 0.68 ± 0.09, 0.59 ± 0.09 and 0.55 ± 0.03, respectively; ApoA1 were 0.58 ± 0.04, 0.83 ± 0.11, 0.92 ± 0.19, 0.71 ± 0.04 and 0.63 ± 0.08, respectively, and the differences among the five groups were statistically significant (F = 25.70, 67.12, 8.94, 11.58; All P < 0.05). P65 the relative expression level were 0.25 ± 0.19, 1.01 ± 0.21, 0.27 ± 0.15, 0.32 ± 0.01 and 0.22 ± 0.11, respectively; AP1 were 0.11 ±0.09, 1.12 ± 0.36, 0.08 ± 0.02, 0.03 ± 0.01 and 0.02 ± 0.01, respectively; JAK2 were 0.76 ± 0.18, 1.11 ± 0.24, 0.34 ± 0.06, 0.42 ± 0.01 and 0.35 ± 0.04, respectively; STAT3 were 0.80 ± 0.33, 1.11 ± 0.27, 0.68 ± 0.22, 0.77 ± 0.06 and 0.68 ± 0.19, respectively, the difference between the 5 groups were statistically significant (F = 43.77, 85.19, 37.22, 17.63; All P < 0.05). Among them, the relative expression levels of PPAR-γ, FATP3, ApoA1, p65, AP1, JAK2 and STAT3 in the infection control group were higher than those in the healthy control group, while those in rosiglitazone group and bexarotene group were lower than those in the infection control group (all P < 0.05). Conclusion A lung injury model of rat infected with P. proliferus was established. The rat model demonstrated that PPAR-γ/RXR-α signalling pathway plays an important role in the mechanism of lung injury caused by P. proliferus infection, and regulation of this signalling pathway by drugs may exert anti-inflammatory effects by inhibiting the NF-κB and JAK/STAT signalling pathways, thereby, reducing the extent of lung injury severity.

Characterization of the mitochondrial genome and phylogenetic implication of Schistosoma japonicum featured with “nocturnal cercarial emergence”
WANG Ning, PENG Hanqi, GAO Changzhe, CHENG Yuheng, LYU Dabing
2023, 41(6):  699-707.  doi:10.12140/j.issn.1000-7423.2023.06.006
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Objective This study aimed to determine the complete mitochondrial genome sequence of Schistosoma japonicum featured with nocturnal cercarial emergence pattern and to explore its phylogenetic relationship with other S. japonicum geographical isolates from mainland China, providing new reference data for further study on population genetic structure and genetic diversity. Methods Field Oncomelania hupensis snails were collected from Shitai County of Anhui in 2020, and the infected snails were separated in the laboratory. ICR mice were infected with cercariae shed from the infected snails using abdominal patch method. The worms were obtained by perfusion from hepatic portal and mesenteric vein and randomly sampled for sequencing. The worm genomic DNA was extracted with blood/tissue DNA extraction Kit, and sequenced at paied-end using Illumina NovaSeq 6000 sequencing platform. The mitochondrial genome was assembled using GetOrganelle tool, and its structural features and base composition were analyzed using MEGA 11 software. The phylogenetic tree was constructed using the neighbour-joining (NJ) and maximum likelihood (ML) method based on 17 published mitochondrial genome sequences of S. japonicum from other parts of China. Results The complete circular mitochondrial genome of S. japonicum with nocturnal cercarial emergence pattern isolated from Shitai County of Anhui was 14 085 bp in length(Genbank accession no. ON637109), encoding 36 genes, consisting of 12 protein-coding genes (PCGs), two ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, with the AT content of 71.35%. There were two gene-overlapping regions and 29 intergenic regions in the mitochondrial genome. Among the 22 tRNA genes, except for trnaC and trnaS1 genes, which missed the dihydrouridine (DHU) arm, the rest were able to form a typical cloverleaf secondary structure. Among the PCGs, the stop codon of cytb, nad4, nad2 and nad6 was TAA, which differed from other geographical strains in China with TAG as the stop codon. Phylogenetic trees constructed based on NJ and ML had similar topologies. The isolated strain of S. japonicum featured with “nocturnal cercarial emergence” in Shitai County, forms a single strain. S. japonicum strains from Taiwan, China and other regions of Chinese Mainland are separated to form one branch. Conclusion The stop codons of S. japonicum isolate from Shitai area of Anhui Province differed from those of other geographical isolates in China. Additionally, based on the phylogenetic analysis, the isolate featured with nocturnal cercarial emergence exhibited a more distant relationship with other regional isolates, showing greater

Prevalence and influencing factors of Enterobius vermicularis infection among kindergarten children in Jiangsu Province from 2019 to 2022
MAO Fanzhen, LI Xuecheng, XU Xiangzhen, DAI Yang, LIU Yaobao, CAO Jun
2023, 41(6):  708-717.  doi:10.12140/j.issn.1000-7423.2023.06.007
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Objective To investigate the current status, trend of changes and influencing factors of Enterobius vermicularis infection in kindergarten children in Jiangsu Province, in order to provide the scientific basis for adjustment and improvement of enterobiasis control strategy. Methods From 2019 to 2022, at least one county (city, district) was selected from the southern, central and northern parts of the Province as survey sites, from each of which two kindergartens (one rural type and one urban type) with a size over 200 children each were selected for cluster sampling the participants. In each kindergarten, no less than 200 chidren separately from senior, middle, and junior classes participated in the suvey. Anal swab method was used to detect E. vermicularis eggs by microscopy. Children’s parents underwent questionnaire survey on basic information of children, hygienic habits related to E. vermicularis infection, family background, and knowledge awareness of enterobiasis. Results A total of 15 669 children were examined in 21 counties (districts, cities), and the overall prevalence rate of pinworm infection was 0.7% in Jiangsu Province from 2019 to 2022. The annual prevalence was 1.2% (44/3 678), 0.5% (13/2 568), 0.7% (27/3 802) and 0.4% (22/5 621), with a significant decrease detected among the years (χ2 = 14.46,P < 0.05). The annual prevalence of rural kindergarten was 2.1% (41/1 981), 0.2% (2/833), 1.0% (8/824) and 0.6% (19/3 271), with a significant decrease detected among the years (χ2 = 17.34,P < 0.05). The annual prevalence of urban kindergarten was 0.2% (4/1 697), 0.6% (10/1 635), 0.7% (20/2 987) and 0.2% (6/2 650), with no significant difference detected among years (χ2 = 0.04, P > 0.05). The overall prevalence of South Jiangsu, Central Jiangsu, and North Jiangsu from 2019 to 2022 was 1.0% (56/5 698), 0.5% (32/6 135), and 0.5% (22/4 136). The prevalence among children of the senior class was 1.8% (23/1 266), 0.8% (8/998), 1.4% (16/1 167), and 0.6% (10/1 796) from 2019 to 2022, respectively, with a significant decrease detected among years (χ2 = 8.01, P < 0.05), which were higher than that among children of the middle and junior class. The prevalence among children of middle class was 1.0% (13/1 325), 0.5% (4/871), 0.5% (9/1 687), and 0.5% (10/2 063) from 2019 to 2022, respectively. The prevalence among children of junior class was 0.8% (9/1 087), 0 (0/699), 0.3% (3/948), and 0.3% (5/1 762) from 2019 to 2022, respectively. The sucking-finger habit accounted for 20.8%, 17.4%, 20.5%, and 19.4% from 2019 to 2022, respectively. The habit of sucking toys/pens accounted for 12.5%, 9.3%, 10.1%, and 12.6% from 2019 to 2022, respectively. A total of 61.9%, 76.0%, 78.5%, and 70.3% (from 2019 to 2022, respectively) parents reported that they knew enterobiasis, with a significant increase detected among the years (χ2 = 202.86, P < 0.05). Being older (OR = 1.447, 95% CI = 1.186-1.764, P < 0.05), a low frequency of laundry (OR = 1.121, 95% CI = 1.002-1.225, P < 0.05), and rural precshool (OR = 0.509, 95% CI = 0.333-0.776, P < 0.05) were risk factors for pinworm infection. Conclusion The prevalence of E. vermicularis infection remained comparatively low in kindergarten children in Jiangsu Province from 2019 to 2022, with a prevalence decreasing trend. However, there exist inadequate hygienic habits in children, and parents showed low knowledge awarness rate on enterobiasis. It is suggested that the factors influencing pinworm infection in kindergarten may include children age, the frequency of washing and sunshining exposure of beddings, and the type of kindergartens.

Surveillance on soil-transmitted helminth infections in Yunnan Province from 2016 to 2022
WU Fangwei, PENG Jia, ZI Jinrong, LI Benfu, YAN Xinliu, CAI Xuan, WANG Zhengqing, XU Qian, LI Jianxiong, YANG Yaming
2023, 41(6):  718-724.  doi:10.12140/j.issn.1000-7423.2023.06.008
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Objective To understand the prevalence and transmission of soil-transmitted helminth (STH) infection in Yunnan Province, providing scientific basis for formulating prevention and control strategies and measures. Methods The surveillance sites were selected in Yunnan Province according to the National Surveillance Program for Clonorchiasis and Soil-transmitted Nematodes (Pilot) from 2016 to 2022. From each district (city), at least one county (city, township) was selected as the surveillance site, which was further divided into 5 sectors (east, west, south, north and central) according to the geographic location. From each sector, one township (town) was selected randomly and an administrative village was selected from the township (town) to conduct the surveillance among permanent residents over 3 years old. No less than 200 residents were cluster sampled from each administrative village and no less than 1 000 residents from each surveillance site. Fecal samples (about 30 g) from the enrolled residents were collected and examined for STH eggs (hookworm, Ascaris lumbricoides, Truchuris trichiura and Enterobius vermicularis) by modified Kato-Katz thick smear method (two slide-reading for each sample). The infection rate and intensity of STH were calculated. The hookworm egg positive samples were further identified for species by tube-filter paper culture method. Children aged 3-9 were additionally examined for E. vermicularis eggs with transparent tape and anal swab method. Chi-square test was used to compare the infection rates. Results A total of 108 221 residents were enrolled in the surveillance from 71 surveillance sites in Yunnan Province and the total STH infection rate was 6.91% (7 475/108 221) from 2016 to 2022. The infection rates of hookworm, A. lumbricoides, T. trichiura and E. vermicularis were 3.75% (4 056/108 221), 1.68% (1 817/108 221), 2.23% (2 408/108 221) and 0.11% (121/108 221) respectively. A total of 1 288 cases of hookworm infection were identified by tube-filter paper culture method. 93.09% (1 199/1 288) of them were infected with Necator americanus, 6.13% (79/1 288) were infected with Ancylostoma duodenale and 0.78% (10/1 288) were co-infected. From 2016 to 2022, the annual infection rates of STH in Yunnan Province were 12.03% (1 606/13 350), 8.67% (1 317/15 195), 7.10% (1 095/15 425), 5.37% (897/16 693), 5.83% (969/16 617), 5.77% (922/15 967) and 4.47% (669/14 974) respectively. The differences between the infection rates among different years were statistically significant (χ2 = 880.360, P < 0.01). Among the 16 prefectures (cities) surveyed, the highest STH infection rate was found in Xishuangbanna (26.21%, 1 312/5 006) and the lowest was found in Lijiang City (0.15%, 6/4 041). The differences between the infection rates among different prefectures (cities) were statistically significant (χ2 = 8 081.151, P < 0.01). The STH infection rates in male and female were 6.35% (3 199/50 409) and 7.40% (4 276/57 812) respectively, with a significant difference (χ2 = 46.198, P < 0.01). Among the different age-groups, nationalities, occupations and educational levels, the highest infection rates were found in the group of 10 to 19 year-old (8.56%, 1 249/14 592), Bulang ethnic group (30.20%, 778/2 576), students (7.96%, 1 638/20 588) and illiteracy or semi-illiteracy group (10.06%, 1 078/10 719), with the significant differences (χ2 = 139.432, 8 546.389, 324.218, 833.291; all P < 0.01). Conclusion From 2016 to 2022, the STH infection rate in Yunnan Province showed a decreasing trend, but remained at a comparatively high level in some areas. It is imperative to continuously strengthen comprehensive control of STH for the key population including the ethnic groups near the border areas, farmers and the elderly.

Prevalence and influence factors of Blastocystis infection in residents of Dong ethnic group in Southeast Guizhou Province
LI Lin, ZHU Aiya, CAI Yuchun, YU Chenghang, CHEN Muxin, TIAN Liguang
2023, 41(6):  725-732.  doi:10.12140/j.issn.1000-7423.2023.06.009
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Objective To investigate the prevalence and influence factors of Blastocystis in the Dong ethnic group in Guizhou Province in 2022. Methods A cross-sectional survey was conducted through questionnaire investigation in the residents of Mangling Village and Diqing Village in Liping County, Dong ethnic group region, southeast Guizhou Province in 2022. The survey collected general demographic information, hygienic habits, evidence of gastrointestinal symtoms. In addition, fecal samples were collected to extract Blastocystis DNA from which the SSU rRNA gene fragment was amplified by PCR. The positive ampicon was sequenced, and the results were aligned by BLAST in the GenBank database to determine the subgenotype of Blastocystis. The MEGA7.0 software was used to construct phylogenetic tree based on the Blastocystis SSU rRNA gene sequence by neighbor-joining method. Chi-square test and Fisher exact probability method were used to analyze the influencing factors of Blastocystis infection in the residents included in this study. Results A total of 329 village residents were surveyed, including 200 females (60.8%) and 129 males (39.2%), with an average age of 47.95 ± 20.39 years, of which 98.5% (324/329) were of Dong ethnic group, and 1.2% (4/329) were of Han ethnic group. PCR results showed that 16 fecal samples amplified Blastocystis SSU rRNA fragments with a size of about 1 100 bp. The overall Blastocystis infection rate was 4.9% (16/329). There was no significant difference in the infection rates in males (3.1%, 3/129) and females (6.0%, 12/200) (χ2 = 1.425, P > 0.05). The infection rate was higher in the age group over 69 years old, 8.1% (3/37), and there was no significant difference in the infection rate among different age groups (χ2 = 3.865, P > 0.05). The Blastocystis infected individuals were all farmers, and the infection rate was 6.1% (16/261), and there was no significant difference in the infection rate between different occupations (χ2 = 3.157, P > 0.05). The infection rate was the highest in households with 6 or more people (12.7%, 7/55), and the difference in infection rate between different household sizes was statistically significant (Fisher = 13.957, P < 0.01). There were 4 genotypes (ST2, ST3, ST6, ST10) infecting the residents of Mangling Village and Diqing Village. Among them, sub-genotype 6 is the most common, accounting for 50.0% (8/16), followed by sub-genotype 3, accounting for 25.0% (4/16). BLAST comparison analysis showed that the similarity between the positive sequences of 16 Blastocystis and the reference sequences was more than 90%. The phylogenetic tree results also showed that each gene subtype clustered with its reference sequence, and the parentage was high. Univariate analysis showed that drinking water source, family size, daily stool frequency and stool traits have significant differences in Blastocystis infection (χ2 = 8.778, 9.552, 7.384, 15.680; P < 0.05). The results of multivariate analysis showed that drinking unboiled water [OR = 18.347, 95% CI: (3.754, 89.673)] was a risk factor for Blastocystis infection in rural residents. Conclusion The overall infection rate of Blastocystis among Dong ethnic residents in southeast Guizhou Province is comparatively, with the main subgenotype ST6. Drinking unboiled water is the main risk factor for infection with Blastocystis in rural residents.

Infection status and genetic variation analysis of Haemonchus contortus in sheep in Shanxi Province
MA Zhiya, XIE Shichen, HE Yuanhui, GAO Wenwei, LIU Qing, ZHU Xingquan, ZHENG Wenbin
2023, 41(6):  733-738.  doi:10.12140/j.issn.1000-7423.2023.06.010
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Objective To understand the prevalence and genetic variation of Haemonchus contortus in sheep in Shanxi Province. Methods Fecal samples were collected from sheep in Shanyin County in Northern Shanxi, Qi County in Central Shanxi and Jishan County in Southern Shanxi for extraction of genomic DNA, of which the partial sequence of H. contortus second internal transcribed spacer 2 (ITS-2) was amplified using H. contortus specific primers by PCR and sequenced to identify the parasite species by gel electrophoresis and evaluate the H. contortus prevalence in sheep in different regions of Shanxi Province. The full-length sequence of ITS of H. contortus was amplified by PCR using conserved primers from ITS-2 positive samples and was sequenced. The ITS-2 sequences and ITS full-length sequences of H. contortus obtained in this study were aligned with the corresponding sequences available in GenBank to examine the base mutation site by DNAMAN 9.0. The base content of H. contortus ITS sequence was calculated using DNAstar 7.1 software. Moreover, the sequence homolohy was analyzed on 18 ITS full-length sequences of H. contortus from different regions and hosts, which were available in GenBank (n = 18), and the 2 ITS full-length sequences obtained in this study. Based on the ITS full-length sequence, phylogenetic tree was constructed using maximum likelihood method to analyze the genetic variation in H. contortus. Results A total of 401 sheep fecal samples were collected, of which 69 samples were successfully amplified with the target ITS-2 fragment (17.2%). The ITS-2 sequences obtained in this study showed 100% sequence identity with that of H. contortus (accession number OQ674251) and were identified as H. contortus. The positive rate of H. contortus was 47.4% (64/135), 3.0% (5/169) and 0 (0/97) in sheep in Shanyin County, Jishan County and Qi County of Shanxi Province, respectively. In addition, the complete ITS sequences of H. contortus were successfully amplified from 4 H. contortus-positive fecal samples, showing two genotypes with a single nucleotide variation at the 226 bp position of ITS-1 locus. Based on the sequence identity analysis of the complete ITS sequences of H. contortus, it was found that the variation rate between the two genotypes identified in this study was only 0.1%, and the variation rate of the 18 sequences from different regions of the world and hosts were 0-2.6%. The analysis of ITS sequences and phylogenetic tree showed that the sequences identified in this study, OP518297 and OP518298, were on the same branch with sequences from the United States (accession number EU086378), Laos (accession number AB908961) and other regions of China (accession number HQ844231), and there was no significant difference among ITS sequences of H. contortus from different regions of the world and hosts. Conclusion The H. contortus infection rate in sheep in the surveyed regions of Shanxi Province was comparatively high, while the genetic variation rate of the parasite detected was low, which showed no significant difference from those of H. contortus found in different regions and hosts.

The molecular biology of Ixodes canisuga and morphology of male ticks on the red fox body surface in Xinjiang
LI Fengshi, ZHAO Shanshan, TAN Wenbo, WURELIHAZI Hazihan, GU Xinli, WANG Suwen, LIU Gang, WANG Yuanzhi
2023, 41(6):  739-743.  doi:10.12140/j.issn.1000-7423.2023.06.011
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Objective To identify suspected ixodid tick species from the body surface of wild red fox (Vulpes vulpes) carcasses in Xinjiang using morphological and molecular biological methods. Methods The ixodid ticks were collected from the body surface of wild red fox carcasses in Bortala Mongolian Autonomous Prefecture and Changji Hui Autonomous Prefecture of Xinjiang in 2018—2019 and observed morphologically with stereomicroscope. Five representative adult ticks (3♂, 2♀) were selected and the mitochondrial gene fragment 16S rDNA was amplified by PCR and sequenced, subsequently, sequences alignment was conducted by BLAST, and the phylogenetic tree was constructed by the maximum likelihood method and homology analysis was performed by MEGA 7.0 software. Results A total of 38 ticks were collected with 36 females and 2 males. The morphological observation and identification showed that the ticks basis capituli was obvious, oval in the pore area, palps and hypostome are long and thin, not closely arranged. The length of the hypostome was about equal to the width of the basis capituli; the peritreme was round and the center was sunken, the scutum are no festoon, no piebaldness and no markings and no eyes in the piebaldness. The foot surface was covered with brown-red enamel, which was relatively thin. The body surface is covered with hairs. The front half of the female tick scutum was covered by the shield, the genital apron is on the edge of genital aperture. The scutum of the male tick was completely covered by the shield, and the anal groove was of anterior groove type, the anal lateral plate and the anal plate were visible; which is consistent with the male I. canisuga morphological characteristics. PCR results showed that the target band of 460 bp was amplified from the DNA of all 5 ticks. Sequence analysis revealed that all the five representative ticks had the same 16S rRNA sequence. The sequence homology with I. canisuga (GenBank accession number: MG651937) from Tarim Basin, Xinjiang, China was 99.47%. The sequence homology with I. canisuga (GenBank accession number: KY962074) from the United Kingdom, France and Germany was 99.20%. Phylogenetic analysis indicated that the representative ticks were clustered to I. canisuga originated from Europe and Xinjiang Province in China, and close to I. lividus (GenBank accession number: KJ414534) and I. arboricola (GenBank accession number: KJ414461). Conclusion Using morphological and molecular biology approaches, I. canisuga was identified from the ticks found on the body surface of Xinjiang red fox, which is highly homologous with the species from Europe.

Analysis on the sources of imported malaria risk in China based on international trade relations
YANG Shuo, XIA Shang, YAN Shuning, XUE Jingbo, SHI Benyun, HAO Yuwan, LI Mengru, LIANG Jiarui, XIA Zhigui, ZHENG Bin
2023, 41(6):  744-748.  doi:10.12140/j.issn.1000-7423.2023.06.012
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Objective To explore the cross-border population mobility between China and malaria-endemic countries worldwide under normal conditions and evaluate the risk sources of imported malaria in China to provide a methodological basis for proactive surveillance and early warning of imported malaria. Methods Malaria prevalence data from global 81 malaria-endemic countries in 2019, the data of Chinese customs import and export of goods, and relevant social, economic, and demographic data were collected to constructe gravity model for cross-border population mobility driven by international trade relations, and assess the risk classification of imported malaria from malaria-endemic countries worldwide. Results The 81 malaria-endemic countries are mainly located in Africa (45 countries, accounting for 55.56%) and Asia (17 countries, accounting for 20.99%). Countries with high malaria endemicity are primarily in sub-Saharan Africa, South Asia, and Southeast Asia. China has trade relationships with all the malaria-endemic countries, accounting for 31.03% of the total trade volume. The model analysis shows that the primary imported malaria risk from countries in central and eastern Africa, South Asia, Southeast Asia, and central-east Latin America. The risk of imported malaria from southern Latin America and Oceania is relatively low. Among the 81 malaria-endemic countries, the three countries with the highest risk of imported malaria to China are India in South Asia, Brazil in South America, and Nigeria in Africa. Conclusion As international exchanges and personnel mobility return to normal, the malaria-endemic countries in central and southern regions of Africa, midwestern Latin America and Southeast Asian are high risk source of imported malaria, thus, it is imperative to exert focusing surveillance and preventive control for the inbound personnel from these risk-regions.

REVIEWS
Research progress on the regulation of miRNA in the infection of apicomplexan parasites
MA Yue, ZHAO Baocai, ZHOU Jiali, HU Junhao, ZHAO Hongxi
2023, 41(6):  749-755.  doi:10.12140/j.issn.1000-7423.2023.06.013
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miRNAs are abundant small non-coding single-stranded RNAs in animals, including humans, that play crucial roles in regulating cellular processes such as cell energy metabolism, proliferation, apoptosis, tumorigenesis, and viral invasion. Apicomplexan parasites are strictly intracellular parasitic protozoa that exhibit host specificity and infect a wide range of hosts. Recent research has revealed the significant regulatory roles of miRNAs during the infection process of apicomplexan parasites. miRNAs serve as novel biomarkers for monitoring the occurrence, development, diagnosis, and prognosis of parasitic diseases and act as novel regulators involved in the interplay between host cells and parasites. This interaction affects the normal physiological processes of host cells and consequently impacts the overall functionality of the organism. This review article summarizes the regulatory mechanisms of miRNAs during apicomplexan parasite infections, aiming to provide valuable insights for exploring preventive, diagnostic, and therapeutic approaches against apicomplexan parasitic diseases.

SHORT COMMUNICATIONS
Phylogenetic analysis of Angiostrongylus cantonensis from Achatina fulica based on COⅠ sequence
GAO Lanlin, XIE Hanguo, LIN Chenxin, JIANG Dianwei, CAI Wuwei, ZHENG Dan
2023, 41(6):  756-759.  doi:10.12140/j.issn.1000-7423.2023.06.014
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The genetic evolution of Angiostrongylus cantonensis infected by the Achatina fulica in Fuzhou was analyzed, and the phylogenetic relationship between A. cantonensis isolates from 5 continents was discussed. The A. fulica were collected from the garbage dump, grass and vegetable fields near the residential areas of major parks in Fuzhou City. The A. cantonensis stage Ⅲ larvae were collected by homogenizing method and identified by morphology. The larval DNA was extracted, and the cytochrome C oxidase subunit Ⅰ (COⅠ) sequence was amplified by PCR. After sequencing, BLAST comparison was performed using the GenBank database. The phylogenetic tree was constructed by the neighbour-joining method using MEGA software with A. vasorum as an outgroup. A total of 103 A. fulica were collected, and 19 of them were infected with stage Ⅲ A. cantonensis larvae. The infection rate was 18.4%. A total of 1 081 A. cantonensis stage Ⅲ larvae were detected. A 450 bp band was amplified by PCR. BLAST results showed that the COⅠ sequence obtained in this study was 98.2% consistent with that of A. cantonensis (GenBank accession number: AB684364.1). Phylogenetic tree analysis showed that the sequences obtained in this study converged on an evolutionary clade with the sequences of the Fujian isolates. The isolates were further clustered with the North America isolates (GenBank accession number: MH069731.1), South America isolates (GenBank accession number: MW390969.1), Oceania isolates (GenBank accession number: KU532144.1) and Europe isolates (GenBank accession number: MN227185.1) into a branch. Therefore, the Fuzhou isolate still belongs to the Asian geographical strain, and the genetic differentiation is stable.

Effect of Clonorchis sinensis infection on hepatic fibrosis and immune regulation in mice
ZHAO Lei, LI Jia, MO Gang, LI Chun, HUANG Guoyang, PENG Xiaohong
2023, 41(6):  760-765.  doi:10.12140/j.issn.1000-7423.2023.06.015
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In order to further understand the liver fibrosis and immune regulatory changes in mice infected with Clonorchis sinensis, BALB/c mice were randomly divided into 2 groups with 30 mice in each group. The infected group were administered 200 μl normal saline with 150 C. sinensis metacercariae through oral gavage and the control group received 200 μl normal saline. Six mice were randomly selected for dissection at 1, 2, 4, 8 and 16 weeks. The paraffin sections of the liver tissue were stained with hematoxylin-eosin (HE) and Masson to observe the progression of fibrosis. The relative transcription level and actin alpha 2 (ACTA2) expression in the liver were detected by qRT-PCR and western blotting separately. The spleen tissue were weighed for calculating the spleen index (spleen weight per 10 g weight, mg/10 g). The flow cytometry was performed to detect the dynamical changes of CD4+ T cells, helper T1 (Th1) cells, Th2 cells, Th17 cells and regulatory T (Treg) cells in spleen. The secretion levels of tumor necrosis factor (TNF), interleukin-6 (IL-6), interferon-γ (IFN-γ), IL-2, IL-4, IL-10, and IL-17A were quantified by cytometric bead array. The data was analyzed by GraphPad Prism 9.0. Two-way ANOVA was used for groups comparison and t test was used for pairwise comparison. The results showed that the mice weight decreased and spleen index increased after infected with C. sinensis and the inflammatory lesions were visible in liver tissue. HE staining showed that C. sinensis larvae and inflammatory cells could be found around the hepatobiliary duct at week 1 after infection. Masson staining revealed that the relative area of collagen fiber deposition was higher than the control group after infection (F = 20.190, P < 0.05). The qRT-PCR results showed that the hepatic ACTA2 relative transcription level of the infected group were higher than the control group at week 2, 4 and 16 after infection (t = 2.042, 2.475, 1.634; all P < 0.01) but lower than the control group at week 8 (t = 2.758, P < 0.05). Western blotting results showed that the hepatic ACTA2 protein relative expression level of the infected group were higher than the control group (F = 3.225, P < 0.01). The flow cytometry results showed that the percentage of splenic CD4+ T cells decreased to (13.4 ± 1.8)% at week 2 after infection and increased to (22.4 ± 1.5)% at week 16. The percentage of Th1 cells increased to (16.9 ± 5.3)% at week 2 and decreased to (3.9 ± 2.6)% at week 16. The percentage of Th2 cells decreased to (2.3 ± 0.6)% at week 4 and increased gradually after week 4. The percentage of Th17 cells increased to (5.3 ± 3.4)% at week 8 and decreased to (2.4 ± 1.4)% at week 16. The percentage of Treg cells decreased to (7.3 ± 1.5)% at week 4 and increased to (13.9 ± 1.2)% at week 16. The serum TNF content of infected group mice increased to (35.16 ± 11.28) pg/ml at week 1 after infection and decreased to (8.98 ± 1.66) pg/ml at week 8. The serum IL-6 content was higher than the control group after infection (t = 2.095, P < 0.05). The serum IL-2 content decreased to (0.09 ± 0.18) pg/ml at week 8 and increased to (3.81 ± 2.79) pg/ml at week 16 after infection. The content of serum IFN-γ, IL-4, IL-10 and IL-17A increased gradually after infection (F = 8.726, 8.068, 6.795, 14.840; all P < 0.05). As a conclusion, the dynamic changes of CD4+ T cells and serum cytokines were closely related to the hepatic fibrosis induced by C. sinensis.

Surveillance on soil-transmitted helminth infections in Gongshan County of Yunnan Province from 2016 to 2021
DUAN Chenli, HE Dongmei, YANG Shu, ZHANG Yanli, LUO Zirong, XIONG Bicai, ZI Jinrong, PENG Jia, WU Fangwei
2023, 41(6):  766-771.  doi:10.12140/j.issn.1000-7423.2023.06.016
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To understand the prevalence and transmission of soil-transmitted helminth (STH) infection in Gongshan County. From 2016 to 2021, according to the National Surveillance Program for Liver Fluke Diseases and Soil-transmitted Nematodes (Pilot). The Gongshan County were selected from five areas (east, west, south, north and central). From each of the areas, a township was randomly selected, and then an administrative village was selected from each township as the surveillance site every year. Over 200 permanent residents above 3 years old were cluster sampled from each administrative village and no less than 1 000 residents were included every year. Fecal samples were collected from the participants for examining and counting STH eggs using the modified Kato-Katz thick smear method to calculate the infection rate. Hookworm egg-positive samples were further identified for species by test tube-filter paper culture method. The eggs of Enterobius vermicularis were detected by the cellophane tape method in children aged 3-9 years. The species and number of infected were recorded, and the infection rate was calculated. The infection rate between groups was compared by the Chi-square test. The fecal samples from 6 175 residents were examined with 928 samples infected with STH. The total infection rate was 15.03% (928/6 175). The infection rates of hookworm, Ascaris lumbricoides, whipworm and Enterobius vermicularis were 3.27% (202/6 175), 7.92% (489/6 175), 8.87% (548/6 175) and 0.19% (12/6 175) respectively. The E. vermicularis eggs were detected by the cellophane tape method in children aged 3-9 years. A total of 823 children were examined and 6 were infected. A total of 202 cases of hookworm infection were identified by the test tube filter method. Among them, 89 cases were infected with Necator americanus, 72 cases were infected with Ancylostoma duodenale and 1 case was infected with both, accounting for 44.06% (89/202), 35.64% (72/202) and 0.50% (1/202), respectively. Minor with hookworm, A. lumbricoides and whipworm infections accounted for 81.19% (164/201, 69.33% (339/489) and 78.29% (429/548) respectively. From 2016 to 2021, the infection rate of STH in Gongshan County was 38.78% (389/1 003), 14.76% (156/1 057), 6.53% (68/1 042), 13.15% (134/1 019), 11.89% (121/1 018) and 5.79% (60/1 036), respectively. There was significant difference in the infection rate among different years (χ2 = 581.98, P < 0.01). Among the five townshipssurveyed, the infection rate of STH was the highest in Dulong River township (32.07%, 398/1 241), and the lowest in Cikai town (9.09%, 112/1 232).The infection rate of STH was statistically significant among different village (χ2 = 359.24, P < 0.01). The infection rate of STH was 11.20% (291/2 598) in male and 17.81% (637/3 577) in female, with significant difference (χ2 = 51.49, P < 0.01). The STH infections were found in all age groups with the highest infection rate in the age group 3 to 9 years old (28.21%, 244/865). People of the Dulong ethnic group had the higher STH infection rate of 30.39% (392/1 290) than people with other ethnic groups. Children possessed the highest infection rate of 27.78% (170/612) compared with people with other occupations. In terms of education, the STH infection rate was highest among preschool child 27.48% (169/615). The infection rate of STH was statistically significant among different agegroups, nationalities, occupations and educational levels (χ2 = 172.16, 310.89, 195.59, 197.59; P < 0.05). The results showed that the infection rate of STH in Gongshan County decreased significantly from 2016 to 2021, but remained at a high level. Combined with the rural revitalization project, the comprehensive control of STH should be continued to strengthen among key populations such as Dulong nationality, children and the elderly.

Clinical features and diagnosis and treatment effect in 47 children cases of visceral leishmaniasis in Gansu Province
CHENG Li, YANG Chengming, ZHANG Ni, XU Xiaonan, LI Yuanxiao, LI Yan, MA Hanwei, LIU Hui, DAI Xingxing
2023, 41(6):  772-775.  doi:10.12140/j.issn.1000-7423.2023.06.017
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A retrospective analysis was performed on 47 children (< 15 years old) with visceral leishmaniasis diagnosed in the Lanzhou University Second Hospital from May 2011 to December 2021 clinical features and diagnostic features. The results showed that all 47 cases had a history of living in endemic areas. Among them, 26 cases were boys (55.3%), which is than 21 cases of girls (44.7%). Most cases were of infants under 3 years old (59.6%). The number of cases was the highest in summer, with 17 cases (36.2%), and sporadic in the rest of the month. The main clinical manifestations were recurrent fever (44 cases, 93.6%), anemia (45 cases, 95.7%), abdominal distension (35 cases, 74.5%), accompanied by spleen enlargement (38 cases, 80.9%), liver enlargement (34 cases, 72.3%) and lymph node enlargement (16 cases, 34.0%). The main laboratory tests were anemia (45 cases, 95.7%), pancytopenia (31 cases, 66.0%) and elevated inflammatory index (43 cases, 91.5%). The rK39 immunochromatographic test was performed for 47 cases, of which 46 cases (97.9%) were positive for Leishmania antibody. Thirty-eight patients underwent bone marrow biopsy smear examination, of which 23 (60.5%) found Leishmania-Donovan bodies. A total of 45 cases were treated with antimony, of which 36 cases were cured after standard antimony treatment (150-200 mg/kg antimony sodium gluconate, divided into 6 days). Significant improvement was seen in 5 cases after 3-6 days of additional treatment, and the effective rate of antimony was 91.1%. One patient died, and three patients had relapse, with a recurrence rate of 6.7%. It is suggested that children with a history of living in endemic areas and meeting the clinical manifestations of visceral leishmaniasis should be diagnosed as early as possible by rK39 immunotomographic test, combined with bone marrow smear examination and standard antimony treatment. When standard antimony treatment is ineffective, additional antimony treatment days are recommended.

CASE REPORTS
A case of visceral leishmaniasis misdiagnosed as liver cirrhosis induced by hepatitis C
JIA Xiaoxia, DENG Chunqing
2023, 41(6):  776-779.  doi:10.12140/j.issn.1000-7423.2023.06.018
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The individual was a 74-year-old farmer from Pingding district of Yangquan City, Shanxi Province. In January 2023, he went to the local hospital due to cough, expectoration and general fatigue. He was diagnosed with mycoplasma, SARS-Cov-19 and other pulmonary infections with mild anemia. The condition temporarily improved after receiving ongoing oxygen therapy and antimicrobial treatment. The patient continued to experience repeated systemic fatigue and was treated in multiple hospitals for over 6 months. On July 10th, he went to the Department of Infectious Diseases at the First Hospital of Shanxi Medical University for treatment. Admission examination: The patient had intermittent low fever, anemic appearance, skin pigmentation from scratching, ascites, soft abdomen and distended, hepatomegaly, splenomegaly, mild lower limb edema. Blood tests showed a leucocyte count of 2.4 × 109/L, hemoglobin 62.0 g/L, red blood cell count of 2.09 × 1012/L, platelet count of 94 × 109/L, and hepatitis C virus antibody positive, abdominal CT noted cirrhosis, hepatomegaly, splenomegaly, while abdominal ultrasound revealed ascites, and liver transient elastography was 24.7, the initial diagnosis was hepatitis C virus cirrhosis. Bone marrow puncture and smear analysis revealed toxic granules in neutrophil cytoplasm and the absence of Leishmania. During a reexamination on July 17th, ELISA returned negative for hepatitis C virus antigen and antibody, and fluorescence quantitative PCR detected hepatitis C virus nucleic acid concentration below 1.0 × 103 IU/ml, implying misdiagnosis initially. The patient had a history of multiple blood transfusions for anemia and had multiple contact histories with street dogs. The history of sandfly bites was unclear. The rK39 immunochromatography strip result and subsequent high-throughput nucleotide sequencing of pathogenic microorganisms identified 4 101 Leishmania DNA sequences, including 95 Leishmania infantum DNA sequences, confirming leishmaniasis. Following treatment with sodium stibogluconate injection (6 ml/d, 10 d), the temperature remained normal, liver and spleen sizes significantly decreased, and skin lesions improved after 15 days of therapy, involving liver protection, protein and red blood cell transfusion, fluid replacement, and diuresis.