Table of Content

28 February 2026, Volume 44 Issue 1

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EDITORIAL

Dual wheel-driven by technological innovation and precision control: advancing schistosomiasis elimination in China during the 15th Five-Year Plan period

XU Jing, CAO Chunli, ZHOU Xiaonong, LI Shizhu

2026, 44(1):  1-7.  doi:10.12140/j.issn.1000-7423.2026.01.001

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Schistosomiasis is a major infectious disease that was once widely prevalent and caused serious damages in China. Elimination of schistosomiasis is an important component to advance the Healthy China Initiative. During the “14th Five-Year Plan” period, China adhered to a strategy anchored in technological innovation and centered on precision control, establishing a working mechanism featuring “Party and government leadership, multi-sectoral collaboration, social mobilization, and whole-of-society participation”. Through integrated control, precise and science-based interventions, China achieved the target of transmission interruption of schistosomiasis nationwide. The paper systematically summarized the effectiveness of integrating technological innovation with precision control in national schistosomiasis control programme in China during the “14th Five-Year Plan” period and analyzed current risks of schistosomiasis rebound, technical bottlenecks and deficiencies in prevention and control capacities. According to the requirements of the Action Plan to Accelerate the Elimination of Schistosomiasis in China (2023-2030), this paper proposes the dual wheel-driven advancement pathways for schistosomiasis elimination anchored in technological innovation and centered on precision control during the “15th Five-Year Plan” period, including strengthening breakthroughs on core technologies, improving precision control systems, refining long-term working mechanisms, and deepening multi-domain collaboration and coordination, so as to provide the theoretical evidence and practical guidance for achieving the target of schistosomiasis elimination and sustainably consolidating the achievements of schistosomiasis elimination across the country.

ORIGINAL ARTICLES

Epidemiological and clinical characteristics of newly reported advanced schistosomiasis patients in Jiangxi Province from 2019 to 2022

GONG Zhihong, YUAN Min, XIE Huiqun, XU Yun, ZHOU Feng, LUO Anqi, LIU Junpu, HU Fei, LIN Dandan, LI Yifeng

2026, 44(1):  8-14.  doi:10.12140/j.issn.1000-7423.2026.01.002

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Objective To analyze the epidemiological and clinical characteristics of newly reported advanced schistosomiasis (AS) cases in Jiangxi Province from 2019 to 2022, so as to provide insights into optimization of the AS control strategy. Methods The epidemiological and clinical characteristics of newly reported AS patients in Jiangxi Province from 2019 to 2022 were collected by means of a retrospective review, medical record inquiry, and questionnaire surveys, including demographics, history of case diagnosis and treatment, comorbidities and complications, laboratory tests (routine blood tests, blood biochemical tests, coagulation functions, and hepatic fibrosis markers) and ultrasound examinations. All data were input into Microsoft Excel 2016, and all statistical analyses were performed with SPSS 25.0. The χ² test was used for comparisons between groups. Results A total of 518 newly reported AS cases were recorded in Jiangxi Province from 2019 to 2022, and 488 cases were included in this analysis. These AS cases were predominantly found in Shangrao City (45.2%, 234/518), Jiujiang City (31.1%, 161/518), Nanchang City (14.1%, 73/518) and Yichun City (9.6%, 50/518), and the majority of these cases were male (62.5%, 305/488). The subjects had a mean age of (66.61 ± 9.84) years, including 82.2% (401/488) at ages of 60 years and older, 88.5% (432/488) with an educational level of primary school and below, and 77.1% (376/488) with an occupation of farmer. Ascites subtype was the predominant clinical subtype (75.4%, 368/488), and 23.6% (115/488) of the subjects were comorbid with other diseases, with hypertension as the most common comorbidity (41 cases), followed by diabetes mellitus (14 cases). Of all cases, 13.1% (64/488) had complications, with upper gastrointestinal bleeding as the predominant complication (53 cases), and the proportions of etiological treatment and antifibrotic therapy were 91.4% (446/488) and 82.6% (403/488). Laboratory tests showed the highest incidence of abnormal platelet counts in blood cell analysis (59.1%, 117/198), and there was a significant difference in the incidence of abnormal platelet counts among subjects with different clinical subtypes of AS (χ² = 12.398, P < 0.05). Regarding liver function parameters, the highest incidence of abnormal albumin was detected (70.7%, 140/198), and regarding hepatic fibrosis markers, the highest incidence of abnormal procollagen typeⅢ (PCⅢ) was recorded (56.6%, 112/198), with a 58.9% (96/163) incidence of abnormal PCⅢ seen among ascites-subtype cases. Regarding coagulation function indicators, thrombin time prolongation was the most common coagulation disorder (52.5%, 104/198), and the proportion of abnormal activated partial thromboplastin time varied in clinical subtypes (χ² = 13.696, P < 0.05). Ultrasound examinations showed grade Ⅲ hepatic parenchymal fibrosis among 89.2% (415/465) of subjects, 69.9% (325/465) among subjects with ascites, and 62.2% (289/465) among subjects with the inner diameter of main portal vein of > 13 mm. Conclusion Elderly men are predominant among newly reported AS cases in Jiangxi Province, and ascites subtype is the predominant clinical subtype, with hypertension and diabetes mellitus as common comorbidities. Despite multiple etiological and antifibrotic treatments given among selected patients, AS remained to progress to advanced stage, suggesting the need to strengthen early monitoring and interventions among patients with comorbid metabolic diseases to slow down disease progression.

Impact of hepatic E-cadherin overexpression on liver fibrosis in mice infected with Schistosoma japonicum

JIANG Tingting, YU Zhihao, CUI Xuanyin, MO Xiaojin, LI Xian, HU Yuan

2026, 44(1):  15-20.  doi:10.12140/j.issn.1000-7423.2026.01.003

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Objective To investigate the impact of E-cadherin (E-cad) overexpression on hepatic epithelial-mesenchymal transition (EMT) and liver fibrosis induced by Schistosoma japonicum infection. Methods A total of 16 mice were randomly divided into the E-cad group and the control group, and were injected with 2 × 10¹¹ vector genomes of E-cad plasmid or empty plasmid packaged with adeno-associated virus 8 via the tail vein. All mice were infected with S. japonicum cercariae [(20 ± 1) per mouse] via the abdomen 3 weeks post-injection, and liver tissues were collected 6 weeks post-infection. Total RNA was extracted from liver tissues, and the relative transcription levels of target genes were detected with a real-time quantitative PCR (qPCR) assay. Total protein was extracted from liver tissues, and the relative expression of target proteins was detected using Western blotting. Paraffin-embedded liver sections were stained with hematoxylin and eosin (HE) and Masson’s trichrome stains to assess the area of single-egg granulomas and collagen fibers. All statistical analyses were performed using the software GraphPad Prism 9, and differences of means between groups were assessed using t-test. Results The relative mRNA transcription levels of E-cad were 1.15 ± 0.07 and 7.19 ± 2.43 in mouse liver tissues in the control and E-cad group (t = 4.30, P < 0.05), and the relative protein expression of E-cad were 1.03 ± 0.16 and 1.42 ± 0.22 in the control and E-cad group (t = 3.62, P < 0.01), indicating successful modeling of hepatic E-cad overexpression in mice. qPCR assay quantified lower relative mRNA transcription of vimentin (Vim) [(0.60 ± 0.17) vs. (1.37 ± 0.33); t = 3.60, P < 0.05], α-smooth muscle actin (α-SMA) [(0.62 ± 0.19) vs. (1.43 ± 0.35); t = 3.48, P < 0.05], collagenⅠ (Col-1) [(0.31 ± 0.17) vs. (1.78 ± 0.84); t = 3.74, P < 0.05] and collagenⅢ (Col-3) [(0.30 ± 0.13) vs. (1.78 ± 0.45); t = 5.52, P < 0.01] in mouse liver tissues in the E-cad group than in the control group, and Western blotting determined lower relative expression of Vim [(0.22 ± 0.13) vs. (0.71 ± 0.13); t = 4.39, P < 0.01], N-cadherin (N-cad) [(0.37 ± 0.14) vs. (0.65 ± 0.08); t = 2.75, P < 0.05], α-SMA [(0.29 ± 0.19) vs. (0.81 ± 0.13); t = 3.91, P < 0.01], Col-1 [(0.23 ± 0.06) vs. (0.43 ± 0.08); t = 3.99, P < 0.01], Col-3 [(0.17 ± 0.02) vs. (0.50 ± 0.16); t = 3.47, P < 0.05], transforming growth factor-β1 (TGF-β1) [(0.32 ± 0.13) vs. (0.68 ± 0.07); t = 3.92, P < 0.01], Smad2 [(0.57 ± 0.14) vs. (0.87 ± 0.04); t = 2.95, P < 0.05] and Smad3 [(0.54 ± 0.10) vs. (0.82 ± 0.06); t = 4.61, P < 0.05] in mouse liver tissues in the E-cad group than in the control group. HE staining showed that the area of single-egg liver granulomas were [(34.10 ± 6.94) × 10⁴] and [(12.76 ± 3.16) × 10⁴] μm² in the control and E-cad groups (t = 7.41, P < 0.01), and Masson staining showed that the areas of collagen fibers with single-egg liver granulomas were [(8.50 ± 2.36) × 10⁴] and [(3.65 ± 0.90) × 10⁴] μm² in the control and E-cad groups (t = 5.01, P < 0.01), respectively. Conclusion Hepatic E-cad overexpression alleviates EMT and liver fibrosis induced by S. japonicum infection in mice, potentially through inhibition of the TGF-β1/Smad signaling pathway.

Surveillance of human soil-transmitted nematode infections in Yunnan Province, 2024

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

2026, 44(1):  21-27.  doi:10.12140/j.issn.1000-7423.2026.01.004

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Objective To investigate the prevalence and epidemiological characteristics of human soil-transmitted nematode infections in Yunnan Province, so as to provide insights into formulation of the soil-transmitted nematodiasis control strategy in the province. Methods A total of 17 counties (cities, districts) were selected as surveillance points using a stratified cluster random sampling method according to the requirements of the National Scheme for Surveillance of Liver Fluke and Soil-transmitted Nematodiasis (Trial) (with Lancang County as a fixed surveillance point). Each surveillance point was divided into five eastern, western, southern, northern, and central areas according to geographical locations, and one administrative village was sampled from each area, followed by 200 local permanent residents at ages of 3 years and older sampled for each administrative village as surveillance subjects using a cluster sampling method, with at least 1 000 individuals surveyed in each surveillance point. Subjects’ stool samples were collected, and detected for soil-transmitted nematode infections and egg counts with the improved Kato-Katz thick smear method (two slides for each stool sample). The prevalence and intensity of soil-transmitted nematode infections were calculated, and differences of prevalence were compared with chi-square test. Pinworm eggs were additionally detected among children at ages of 3 to 9 years with a cellophane tape test. Five households were randomly sampled from each administrative village at fixed surveillance points, and one farmland or vegetable garden soil sample was collected from each household to detect hookworm larvae and human Ascaris lumbricoides eggs in soil samples. Results A total of 17 554 residents were detected for soil-transmitted nematode infections in 17 counties (cities, districts), and the prevalence of soil-transmitted nematode infections was 2.81% (493/17 554). The highest prevalence of soil-transmitted nematode infections was seen in southern Yunnan Province (5.27%, 384/7 288) (χ² = 297.64, P < 0.05), and the prevalence of soil-transmitted nematode infections was higher in border counties (11.69%, 370/3 166) than in non-border counties (0.85%, 123/14 388) (χ² = 1 115.42, P < 0.05). As of surveillance points, the highest prevalence was recorded in Lancang County (23.87%, 263/1 102) (χ² = 2 094.64, P < 0.05). The prevalence of soil-transmitted nematode infections was 2.60% (214/8 237) in males and 2.99% (279/9 317) in females, and the highest prevalence was observed among individuals at ages of 60 to 69 years (3.72%, 99/2 658) (χ² = 47.97, P < 0.05). As of ethnicity, Jingpo (1/2), Lahu (27.68%, 155/560), Hani (14.74%, 102/692), and Wa (12.66%, 10/79) ethnic populations had the four highest prevalence of soil-transmitted nematode infections (χ² = 1 828.99, P < 0.05), and in terms of occupations and educational levels, the highest prevalence was seen among farmers (3.37%, 457/13 563) (χ² = 71.94, P < 0.05) and illiterate individuals (6.71%, 89/1 327) (χ² = 143.84, P < 0.01). The prevalence of hookworm, A. lumbricoides, Trichuris trichura, and pinworm infections was 2.47% (434/17 554), 0.05% (9/17 554), 0.26% (45/17 554), and 0.07% (13/17 554), respectively. The prevalence of mild and moderate infections was 97.00% (421/434), 8/9, and 88.89% (40/45), and 1.38% (6/434), 1/9, and 11.11% (5/45) among individuals with hookworm, A. lumbricoides, T. trichura infections, respectively, and the prevalence of severe infection was 0.38% among individuals with hookworm infections (0.38%, 6/434), while no severe infections were found among individuals with A. lumbricoides or T. trichura infections. A total of 25 soil samples were detected for hookworm larvae and A. lumbricoides at fixed surveillance points, and the detection of hookworm larvae was 44.00% (11/25); however, no A. lumbricoides eggs were detected. In addition, tapeworm, Trichostrongylus orientalis, and Hymenolepis diminuta infections were detected. Conclusion The prevalence of soil-transmitted nematode infections is high in ethnic minority border regions of Yunnan Province, notably hookworm infections. Implementation of integrated soil-transmitted control interventions should be given be a high priority in future parasitic disease control programs in ethnic minority border regions of Yunnan Province.

Spatiotemporal clustering analysis of seropositivity of key foodborne parasites in Dali Bai Autonomous Prefecture from 2019 to 2023

HAO Mingming, LI Rong, CHEN Ran, GAO Yuanyuan, DUAN Xiaoyun

2026, 44(1):  28-34.  doi:10.12140/j.issn.1000-7423.2026.01.005

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Objective To systematically unravel the spatiotemporal distribution and clustering characteristics of seroprevalence of IgG antibodies against six major foodborne parasites in Dali Bai Autonomous Prefecture, Yunnan Province, based on serological testing data, so as to provide insights into precision diseases prevention and control. Methods Serum samples were collected from individuals admitted from Dali Bai Autonomous Prefecture Institute of Schistosomiasis Control from 2019 to 2023, and serum IgG antibodies against pathogens of six major zoonotic parasitic diseases were detected using enzyme-linked immunosorbent assay (ELISA) or colloidal gold immunochromatography, including taeniasis/cysticercosis, angiostrongyliasis, trichinosis, fascioliasis, echinococcosis, and paragonimiasis. Geographic distribution was visualized using the software ArcMap 10.8, and the overall spatial clustering pattern of IgG antibody seropositivity was identified using Moran’s I index within the study area. High-value (hot spots) and low-value (cold spots) clusters were identified with local statistics within the study area, and spatiotemporal scan analysis was performed with the software SaTScan 10.1 based on the Poisson model to identify spatiotemporal clusters of IgG antibody positivity against parasites. Results A total of 3 131 serum samples were tested from 2019 to 2023, and the overall seroprevalence of IgG antibodies against foodborne parasites was 23.3% (728/3 131) in Dali Prefecture, with seroprevalence of 52.5% (139/265), 50.5% (161/319), 26.7% (211/789), 14.6% (140/961), and 9.7% (77/797) from 2019 to 2023, respectively, appearing a tendency towards decline over years (χ² = 387.55, P < 0.01). Single infections were predominant (18.6%, 583/3 131), followed by dual infections (4.2%, 132/3 131). Counties with the three highest overall seropositivity included Yunlong County (53.2%, 58/109), Nanjian Yi Autonomous County (35.5%, 33/93), and Xiangyun County (32.7%, 33/101). The Moran’s I index was all negative each year from 2019 to 2023, ranging from -0.20 to -0.03, indicating no spatial autocorrelation in the seropositivity across years (all P > 0.05), which was consistent with a random spatial distribution pattern. Hotspot areas exhibited a random spatial distribution, with significant hotspots shifting from western regions (Yunlong County, Yangbi Yi Autonomous County) to central regions (Dali City) and then to eastern areas (Midu County) over time. Spatiotemporal scans identified two statistically significant clusters, including Heqing County (2021 to 2022, RR = 6.47, P < 0.01) and Weishan Yi and Hui Autonomous County (2021 to 2022, RR = 5.30, P < 0.01). Parasite IgG antibody-positive samples were primarily derived from farmers (86.1%, 627/728), males (59.1%, 430/728), and individuals at ages of 40 to 59 years (55.6%, 405/728), and no gender-, age- or occupation-specific seroprevalence was seen (χ² = 1.97, 7.73, 10.64, all P > 0.05). Conclusion The overall risk of foodborne parasitic infections declines in Dali Prefecture; however, the spatial pattern appears a complex landscape that is characterized by global randomness, local clustering, and dynamic shifts of hotspots. Differentiated, precision interventions are needed for long-term high-burden areas, anomalous clusters, and dynamic hotspots, and health education and behavioral interventions are prioritized for high-risk individuals, particularly farmers.

Preliminary investigation of the clinical significance of neutrophils in patients with hepatic alveolar echinococcosis

FAN Yichen, ZHANG Chuanshan, HOU Jiao, TANG Zhaoyuan, LU Xuemei, HE Rongdong, JIAYIDAER Humaerhan, SONG Zhihao, KAN Mingxuan, WANG Mingjuan, SUN Li, WEN Hao

2026, 44(1):  35-41.  doi:10.12140/j.issn.1000-7423.2026.01.006

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Objective To investigate neutrophil levels in peripheral blood and liver tissues among patients with hepatic alveolar echinococcosis (HAE), immune microenvironment characteristics, and their associations with the severity of HAE. Methods A total of 24 patients with AE who underwent surgical treatment in Qinghai University Affiliated Hospital from July 2024 to September 2025 (AE group) and 24 healthy controls (HC group) were included. All subjects in the AE group were definitively diagnosed by imaging and pathology. Subjects’ gender, age, neutrophil count, lymphocyte count, platelet count, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were collected, and the neutrophil to lymphocyte ratio (NLR) and systemic immune-inflammation index (SII) were calculated, with AE patients subjected to PMN classification. The correlations between peripheral blood clinical indicators were examined using the software GraphPad Prism 9 in the AE group. Peripheral blood, and surgically resected peri-lesional and distal liver tissues were sampled for hematoxylin and eosin (HE) staining, Masson staining, and immunohistochemical staining, and the proportions of areas stained positive for C-C motif chemokine ligand 2 (CCL2), C-X-C chemokine ligand 1 (CXCL1), and C-X-C motif chemokine receptor 2 (CXCR2) were estimated using the software QuPath-0.5.1, and the associations of CD16⁺ CD66b⁺ neutrophil levels in liver tissues with clinical indicators were examined using flow cytometry. The expression of inflammatory factors interleukin-8 (IL-8), IL-10, tumor necrosis factor alpha (TNF-α), granzyme B (GRZB), CCL2, elastase 2 (ELA2), intercellular adhesion molecule-1 (ICAM-1), gelatinase associated lipocalin (NGAL), matrix metalloproteinase (MMP-9) and myeloperoxidase (MPO) was measured peripheral blood and liver tissues using ELISA, and a correlation network analysis was performed. Results The peripheral blood lymphocyte [(1.60 ± 0.64) × 10⁹/L vs. (1.99 ± 0.62) × 10⁹/L; t = 2.036, P < 0.05] and platelet counts [(229.75 ± 62.20) × 10⁹/L vs. (260.29 ± 40.36) × 10⁹/L; t = 2.018, P < 0.05] were lower in the AE group than in the HC group, and the ALT [(35.54 ± 27.53) U/L vs. (24.17 ± 5.29) U/L; t = 2.319, P < 0.05], AST [(49.21 ± 47.63) U/L vs. (49.20 ± 47.64) U/L; t = 2.604, P < 0.05], NLR [2.06 (range, 1.48 to 3.27) vs. 1.67 ± 0.37; t = 2.317, P < 0.05] and SII [725.29 (range, 312.41 to 810.59) vs. 420.58 ± 44.81; t = 2.051, P < 0.05] were higher in the AE group than in the HC group. Correlation analysis revealed that NLR and SII strongly positively correlated with PMN classification (r = 0.67, P < 0.01), and weakly positively correlated with ALT (P < 0.05). Pathological examinations showed severe tissue structural disorders, hepatocyte coagulative necrosis, and fibrosis in peri-lesional liver tissues relative to distal tissues. Flow cytometry measured higher CD16⁺ CD66b⁺ neutrophil levels in peri-lesional liver tissues (20.49 ± 10.63) than in distal tissues (33.52 ± 11.23) (t = 6.923, P < 0.01), and the CD16⁺ CD66b⁺ neutrophil levels in peri-lesional liver tissues were positively correlated with PMN classification (r = 0.667, P < 0.05). Immunohistochemistry showed higher expression of CCL2 [(50.03 ± 15.30) vs. (40.99 ± 13.78); t = 6.492, P < 0.05], CXCL1 [(72.77 ± 13.45) vs. (56.94 ± 16.23); t = 5.527, P < 0.05], and CXCR2 [(74.49 ± 10.55) vs. (57.76 ± 15.26); t = 3.747, P < 0.05] in peri-lesional liver tissues than in distal tissues. ELISA measured elevated peripheral blood IL-6, IL-10, ELA2, NGAL, MMP-9, MPO, and TNF-α levels, high IL-8, IL-10, GRZB and CCL-2 expression and low NGAL and MMP-9 expression in peri-lesional liver tissues in the AE group. Correlation network analysis revealed strongly positive correlations between MPO and ICAM-1 (r = 0.98), and between MMP-9 and ELA2 (r = 0.88). Conclusion Remarkable infiltration of neutrophils driven by upregulation of chemokine axes (CXCL1, CXCR2, CCL2) is observed around the liver lesions among AE patients. Neutrophils drive cytokine storm and fibrosis remodeling around lesions through the synergistic effect of releasing oxidases and matrix-degrading enzymes. The pathological changes caused by the deterioration of local immune microenvironments determine the severity of AE (PNM classification) and synchronously project the significant increase of peripheral blood systemic inflammatory markers (NLR and SII).

Epidemiological characteristics of malaria in Guangdong Province from 2015 to 2024

ZHANG Jiayi, LU Wencheng, WU De, LIU Jun, LIAO Yuhuang, MAO Qiang, CHEN Lijun, DENG Zhuohui, ZHANG Xianchang, XIA Zhigui, CHEN Jingdiao

2026, 44(1):  42-49.  doi:10.12140/j.issn.1000-7423.2026.01.007

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Objective To analyze the epidemiological characteristics of malaria cases reported in Guangdong Province from 2015 to 2024, so as to provide insights into improving the malaria control strategy and preventing re-establishment of imported malaria. Methods Data on malaria cases and epidemiological case investigation forms for malaria reported in Guangdong Province from 2015 to 2024 were collected from the Infectious Disease Reporting Information Management System and the Parasitic Disease Prevention and Control Information System of Chinese Center for Disease Control and Prevention. The malaria parasite species, origin of acquiring infections, and the temporal, spatial and population distribution characteristics, healthcare-seeking behaviors and diagnosis of malaria cases were statistically analyzed with the software Microsoft Excel 2016 and SPSS 26.0. Results A total of 1 879 malaria cases were reported in Guangdong Province from 2015 to 2024, including 1 871 overseas imported cases, 2 transfusion-transmitted cases, and 6 cases with recrudescent long-latency Plasmodium malariae malaria, and a total of 16 deaths occurred. The number of reported malaria cases appeared an overall tendency towards a rise from 2015 to 2019, and the annual average number of reported malaria cases was all lower during the COVID-19 pandemic (from 2020 to 2022) than the 10-year mean number. Laboratory confirmation showed 1 559 cases (83.0%) with P. falciparum malaria, and the proportion of P. falciparum malaria cases ranged from 80.4% to 86.8% in all malaria cases from 2015 to 2024. Among 1 871 imported malaria cases, 1 785 cases (95.4%) acquired infections in Africa. Malaria cases were reported across Guangdong Province except in Chaozhou City, and were primarily found in the Pearl River Delta region (1 680 cases, 89.4%). The monthly number of malaria cases was 1 to 65, with a median of 15 cases per month. The annual number of malaria cases peaked in June and during the period between August and October. The ratio of male to female cases was 8.8:1, and age distribution peaked at 30 to 39 years (32.5%, 611/1 879) and 40 to 49 years (26.7%, 501/1 879). Malaria cases were reported by 191 institutions, and 78.4% (1 472/1 879) of cases sought healthcare services within 3 days of onset, with a higher proportion seen among P. falciparum malaria cases (79.7%, 1 242/1 559) than among cases with other types of malaria (χ² = 8.17, P < 0.01). The proportion of correct initial diagnosis was 83.1% (1 562/1 879), and the percentages of correct initial diagnosis at city-level and higher, county (district)-level and grassroots healthcare facilities were 90.6% (1 096/1 210), 81.7% (402/492), and 36.2% (64/177), respectively (χ² = 327.04, P < 0.01). The median interval from healthcare-seeking to definitive diagnosis was 1 day, with 86.6% (1 627/1 879) of cases that were definitively diagnosed within 3 days of healthcare-seeking, and the proportion of P. falciparum malaria cases that were definitively diagnosed within 3 days was higher than that of cases with other types of malaria (χ² = 12.44, P < 0.01). Severe malaria was reported in 69 cases (3.7%, 69/1 879), all of which were caused by P. falciparum infection. Conclusion The risk of imported malaria continues to rise in Guangdong Province, with P. falciparum malaria predominating among reported cases. Future control efforts should focus on reinforcement of health education among individuals enter and exit China and improvements in the early-stage diagnostic capability in grassroots healthcare facilities.

Determinants of healthcare-seeking and diagnosis among imported malaria cases in Hangzhou City from 2016 to 2024

ZHENG Caifang, HUO Liangliang, XU Minjie, JIN Xingyi, ZHU Sujuan, LIU Shuai, JIN Quan

2026, 44(1):  50-56.  doi:10.12140/j.issn.1000-7423.2026.01.008

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Objective To investigate the delay in healthcare-seeking and diagnosis of imported malaria cases and identify their determinants in Hangzhou City from 2016 to 2024, so as to provide the scientific evidence for improving the management of imported malaria cases in Hangzhou City. Methods Malaria surveillance data and epidemiological case investigation forms reported in Hangzhou City from 2016 to 2024 were collected from Infectious Diseases Report Information Management System and Parasitic Diseases Prevention and Control Information Management System of Chinese Center for Disease Control and Prevention, including demographic characteristics, disease onset and healthcare-seeking, source of acquiring infections, previous medical history, treatment, and parasite species. All statistical analyses were conducted using the software SPSS 26.0. The determinants of delay in healthcare-seeking and diagnosis were identified using a univariate logistic regression model among imported malaria cases, and the association of delay in healthcare-seeking and diagnosis with severe malaria was examined using a multivariate logistic regression model. Results A total of 282 imported malaria cases were reported in Hangzhou City from 2016 to 2024, including 265 men (94.0%) and 17 women (6.0%), and there were 226 overseas labors (80.1%) and 271 African imported cases (96.1%). Most cases (40.8%, 115/282) initially sought healthcare services in district (county)-level medical institutions, and the misdiagnosis rate of initial healthcare-seeking was 22.7% (64/282). Among all reported cases, most acquired Plasmodium falciparum infections (74.8%, 211/282). The interval between disease onset and initial healthcare-seeking was 0 to 16 days, with 30.1% (85/282) of cases seeking healthcare on the day of disease onset. The interval between initial healthcare-seeking and definitive diagnosis was 0 to 46 days, with 38.6% (109/282) of the cases receiving definitive diagnosis on the day of healthcare-seeking. The proportions of cases with delays from disease onset to initial healthcare-seeking, from initial healthcare-seeking to definitive diagnosis, and from disease onset to definitive diagnosis were 13.1% (37/282), 28.4% (80/282), and 18.4% (52/282), respectively, and the percentages of cases with delay from disease onset to initial healthcare-seeking were 15.5% (9/58), 9.9% (7/71), 6.3% (5/79), and 21.6% (16/74) among cases under 30 years of age, at ages of 30 to 39 years, 40 to 49 years and 50 years and older (χ² = 8.843, P < 0.05). There were significant differences in the proportions of cases with delays from initial healthcare-seeking to definitive diagnosis in terms of sources of acquiring infections (χ² = 3.860, P < 0.05), levels of medical institutions for initial healthcare seeking (χ² = 19.768, P < 0.05), initial diagnosis results (χ² = 113.928, P < 0.05), and infected Plasmodium species (χ² = 13.030, P < 0.05), and there were significant differences in the proportions of cases with delays from disease onset to definitive diagnosis in terms of age groups (χ² = 9.211, P < 0.05), sources of acquiring infections (χ² = 5.554, P < 0.05), interval from disease onset to overseas travel history (χ² = 15.354, P < 0.05), initial diagnosis results (χ² = 54.830, P < 0.05), and infected Plasmodium species (χ² = 31.735, P < 0.05). Univariate logistic regression analysis identified age of 50 years and older as a risk factor for delay in healthcare-seeking (OR = 1.245, 95%CI: 1.085-1.708). Cases with an overseas travel history of more than 6 months prior to disease onset (OR = 3.057, 95%CI: 1.041-8.979), initial diagnosis as other diseases (OR = 29.405, 95%CI: 13.993-61.789), P. vivax infection (OR = 2.717, 95%CI: 1.016-7.266), and infection with other malaria parasite species (OR = 2.810, 95%CI: 1.498-5.273) were more likely to experience delay in diagnosis, and initial healthcare-seeking at province-level medical institutions was a protective factor for delay in diagnosis (OR = 0.135, 95%CI: 0.023-0.800). Of 282 imported malaria cases, there were 11 severe cases and deaths, including 10 P. falciparum malaria cases, and one P. ovale malaria case. Multivariate logistic regression analysis revealed that delay in diagnosis (OR = 6.285, 95%CI: 1.625-24.302) and overall delay (OR = 6.046, 95%CI: 1.491-24.522) were significantly associated with an increased risk of severe malaria. Conclusion There was a delay in healthcare-seeking and definitive diagnosis among imported malaria cases in Hangzhou City from 2016 to 2024, which was mainly attributed to age, place of overseas travel, level of medical institutions for initial healthcare-seeking, and Plasmodium species.

Establishment and validation of a mouse model of infection with Plasmodium berghei chimerically expressing P. falciparum circumsporozoite protein

WANG Jiji, YUAN Yajie, ZHENG Jian, HAN Lu, GYAWU Stephen Baffour, ZHANG Qinglong, FENG Gaoqian

2026, 44(1):  57-63.  doi:10.12140/j.issn.1000-7423.2026.01.009

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Objective To establish a mouse model of infection with Plasmodium berghei (Pb) chimerically expressing P. falciparum circumsporozoite protein (PfCSP), and to evaluate the value of the model in evaluation of vaccine-induced immunoprotective effectiveness, so as to provide a reliable proof-of-concept platform for advancing CSP-based malaria vaccines. Methods PCR assay was employed to characterize and verify the transgenic Pb strain chimerically expressing the Pfcsp gene (PfCSP-Pb strain). The proliferation, morphology, and parasitemia were compared between the PfCSP-Pb and wild-type Pb strains during the erythrocytic stage through in vitro culture for 1 to 5 days and in vivo passage in mice for 1 to 6 days. ICR mice that had been infected with either the PfCSP-Pb strain or the wild-type Pb strain for 3 to 4 days were fed to Anopheles stephensi mosquitoes. The oocysts were observed in mosquito midguts using mercurochrome staining 14 days post-infection with An. stephensi, and green fluorescent protein (GFP) fluorescence signals of sporozoites were detected in mosquito midguts and salivary glands using fluorescence microscopy. Salivary gland sporozoites from Anopheles mosquitoes infected with either the PfCSP-Pb strain or the wild-type Pb strain were collected and intravenously injected into ICR mice via tail vein. Mice were sacrificed 42 hours post-infection, and mouse liver tissues were sampled. Total RNA was extracted from mouse liver tissues, and the relative expression of sporozoite 18S rRNA was detected in mice during the liver stage using real-time quantitative real-time reverse transcription PCR (qRT-PCR) assay. Following passive immunization with the PfCSP-specific mAB317 monoclonal antibody (100 μg) in mice, mice were infected with salivary gland sporozoites from Anopheles mosquitoes infected with either the PfCSP-Pb strain or the wild-type Pb strain. Mouse liver parasite load was detected using qRT-PCR assay to verify the blocking effect of the mAB317 monoclonal antibody on the infection of hepatocytes with sporozoites of the PfCSP-Pb strain. Results PCR assay confirmed the stable integration and expression of the pf-csp gene and the gfp-luciferase reporter gene by the PfCSP-Pb strain. No significant differences were observed in proliferation rates between the PfCSP-Pb strain and the wild-type Pb strain either in in vitro (F = 0.86, P > 0.05) or in vivo (F = 2.18, P > 0.05) during the erythrocytic stage. The PfCSP-Pb strain exhibited identical morphology to the wild-type strain across the ring, trophozoite, and schizont stages, with stable GFP fluorescence protein expression observed in the cytoplasm. Mercurochrome staining revealed that both the PfCSP-Pb strain and the wild-type Pb strain produced a large number of oocysts in the midguts of Anopheles mosquitoes infected with both the PfCSP-Pb strain and the wild-type Pb strain on day 14 post-infection, which contained sporozoite precursor cells, and GFP fluorescence signals were detected in the salivary glands and midgut tissues of Anopheles mosquitoes infected with the PfCSP-Pb strain. In the liver stage, qRT-PCR assay showed that the relative 18S rRNA expression was 1.26 ± 0.49 in the PfCSP-Pb strain infection group, which was comparable to that in the wild-type Pb infection group (1.16 ± 0.67) (t = 0.27, P > 0.05). Following immunization of the mAB317 monoclonal antibody, the relative liver parasite load was lower in the PfCSP-specific monoclonal antibody treatment group (0.25 ± 0.48) than in the control group (1.10 ± 0.51) (t = 2.70, P < 0.05) in mice infected with salivary gland sporozoites from Anopheles mosquitoes infected with the PfCSP-Pb strain, while no significant difference was seen in the relative liver parasite load between the PfCSP-specific monoclonal antibody treatment group (0.92 ± 0.44) and the control group (1.28 ± 0.69) (t = 0.99, P > 0.05) in mice infected with salivary gland sporozoites from Anopheles mosquitoes infected with the wild-type Pb strain. Conclusion This study successfully established a chimeric Pb infection model stably expressing chimeric PfCSP. The PfCSP-Pb strain maintains stable biological characteristics both in vitro and in vivo, and is specifically able to be used to evaluate antibody-mediated immunoprotection against PfCSP, which provides a key technical platform and evaluation tool for preclinical researches on development of P. falciparum malaria vaccines.

Phosphorothioate-dNTP assisted RPA coupled with CRISPR/Cas12a for rapid genotyping of Plasmodium

HUANG Xiao, CHEN Ying, WANG Maoquan, CHEN Yating, LUO Guangcheng

2026, 44(1):  64-71.  doi:10.12140/j.issn.1000-7423.2026.01.010

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Objective To establish a rapid nucleic acid assay for genotyping for Plasmodium (PRCP) based on phosphorothioate dNTPs (dNTPαS) enhancement of the specificity and sensitivity of recombinase polymerase amplification (RPA), combined with the target nucleic acid recognition and signal amplification capabilities of CRISPR/Cas12a. Methods Universal RPA primers were designed with the software Primer Premier 6, and specific gRNAs targeting different species of Plasmodium were designed within the universal primer region using the software SnapGene 6.0. Then, dNTPαS was added to the reaction system to generate an S-RPA amplification reaction, and CRISPR/Cas12a was employed for typing recognition and signal amplification output of the amplification product. The dNTPαS concentration, RPA primer concentration, gRNA concentration, Cas12a concentration, reaction temperature, reaction time, and final Cas12a cleavage time in the PRCP reaction system were optimized in sequence. PRCP was performed with P. falciparum plasmids at concentrations of 10⁸, 10⁷, 10⁶, 10⁵, 10⁴, 10³, 10², 10¹ copies/µL as templates to evaluate its sensitivity, and hepatitis B virus, Babesia, Trypanosoma brucei, influenza A virus, influenza B virus, Mycoplasma pneumoniae, and Chlamydia pneumoniae served as controls to evaluate the specificity, and was conducted with addition of 2 g/L hemoglobin, 0.1 mmol/L triglyceride, and 1 μmol/L bilirubin to evaluate its anti-interference ability. In addition, mixed plasmid samples were used to detect the ability of the PRCP system to distinguish mixed infections, and the consistency was compared between detection of mixed plasmid samples and clinical samples (10 samples of Plasmodium infections and 10 negative samples) with thick and thin blood smears. Results A dNTPαS-assisted RPA assay was established based on 3F3R screened as the universal Plasmodium nucleic acid RPA primer to construct a PRCP system. The optimized parameters for the PRCP system included the optimal proportion of dNTPαS as 70%, the optimal final concentration of primers as 0.50 μmol/L (Rate10 as 676.36), the final concentration of Cas12a as 0.10 μmol/L (Rate10 as 338.28), and the final concentration of gRNA as 0.10 μmol/L (Rate10 as 718.90), and the RPA reaction conditions included 39 ℃ (grayscale value of 32 570 ± 5 045) and 20 minutes (grayscale value of 22 513 ± 156), with Cas12a cleavage for 15 minutes as the detection endpoint (grayscale value 8 624 ± 359). The detection sensitivity of the PRCP system was 100 copies/μL or below, and no cross-reactivity was found with hepatitis B virus, Babesia, T. brucei, influenza A virus, influenza B virus, M. pneumoniae, or C. pneumoniae. The PRCP system was found to resist interference from hemoglobin, triglyceride, and bilirubin, and was able to detect mixed infections. Compared with thick and thin blood smears, the PRCP system showed a 20/20 consistency for detection. Conclusion A rapid, sensitive and specific PRCP assay has been successfully established for rapid genotyping of Plasmodium genes, which provides a novel protocol for early screening and precise diagnosis and treatment of Plasmodium.

Prevalence and phylogenetic analysis of protozoan in Haemaphysalis ticks from Chongming, Shanghai

SHEN Yong, LI Yuanyuan, WANG Ziyi, YANG Limin, HUANG Lirong, LI Zhongqiu, HAN Qingchi, ZHANG Yi, GUO Yunhai, LIU Qin

2026, 44(1):  72-78.  doi:10.12140/j.issn.1000-7423.2026.01.011

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Objective To investigate the prevalence and genetic characteristics of common Apicomplexan protozoan in ticks collected from Chongming, Shanghai, so as to provide insights into management of tick-borne diseases in Chongming area. Methods Free-living ticks were collected using the flagging method in Chongming, Shanghai, from July to September 2023. Following morphological identification, ticks were pooled according to species and developmental stage (one adult per pool, five nymphs per pool, or ten larvae per pool), and genomic DNA was extracted from each pool. Tick species were molecularly identified by PCR amplification of the cytochrome c oxidase subunit 1 (cox1) gene and 16S rDNA gene. The short fragment of the Babesia/Theileria 18S rRNA gene (approximately 400 bp in length) was amplified using nested PCR assay, and positive samples were further subjected to amplification of the long fragment of the 18S rRNA gene (approximately 1 600 bp in length), and positive nested PCR products were sequenced, followed by sequence alignment with BLAST. Genetic distances were calculated among different samples using the software MEGA 11 and phylogenetic trees were created. Differences in the detection of protozoan were tested for statistical significance with chi-square test. Results A total of 622 free-living ticks were collected, including 255 Haemaphysalis longicornis and 367 H. flava. H. longicornis ticks were divided into 83 pools (55 adult pools, 16 nymph pools and 12 larval pools), and H. flava were divided into 43 pools (2 adult pools, 9 nymph pools and 32 larval pools). Nested PCR amplification of the short fragment of the 18S rRNA gene yielded 17 positive pools, including 16 H. longicornis pools (9 adult pools and 7 nymph pools) and one H. flava pool (adult pool). and the long fragment of the 18S rRNA gene was successfully amplified from all 17 positive pools, with sequences deposited under accession numbers of PX453257 to PX453273. BLAST alignments revealed that one gene sequence showed 99.52% identity with the Babesia 18S rRNA gene sequence (MK930513), 7 gene sequences showed 91.53% to 99.94% identity with the 18S rRNA gene sequences of the novel protozoan Colpodella, and 9 gene sequences showed 95.66% to 99.36% identity with the Colpoda 18S rRNA gene sequences. Genetic distance analysis indicated high intraspecific sequence conservation, with genetic distances ranging from 0.003 to 0.147 among 7 Colpodella gene sequences and from 0.009 to 0.060 among 9 Colpoda gene sequences, and the genetic distance between Colpodella and Colpoda ranged from 0.124 to 0.221, indicating substantial interspecific divergence. Phylogenetic analysis revealed that Colpoda and Colpodella were clustered into a large clade, and Babesia and Theileria were clustered into another large clade. The detection rates of Babesia, Colpodella, Colpoda, and protozoan were 1.20% (1/83), 8.43% (7/83), 9.64% (8/83), and 19.28% (16/83) in H. longicornis, and 0 (0/43), 0 (0/43), 2.33% (1/43), and 2.33% (1/43) in H. flava, respectively. The overall detection of protozoan was significantly higher in H. longicornis than in H. flava (χ² = 6.29, P < 0.05). Conclusion There were Babesia, Colpoda, and novel protozoan Colpodella infections in free-living ticks collected from Chongming area, indicating a potential risk of tick-borne diseases.

Epidemiological characteristics and genetic evolutionary analysis of tick-borne piroplasm in selected areas of northwestern China

XIAO Fangyu, LUO Jin, DUAN Deyong, ZHU Yanmin, DIAO Peiwen, HUANG Weixia, ZHANG Yu, WANG Junhong, REN Qiaoyun, GUAN Guiquan, YIN Hong, LIU Guangyuan

2026, 44(1):  79-84.  doi:10.12140/j.issn.1000-7423.2026.01.012

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Objective To investigate the epidemiological characteristics and genetic evolutionary relationships of piroplasm in ticks in selected areas of northwestern China. Methods Ticks infesting the livestock body surface were collected from Zhangjiachuan County, Zhuanglang County, Chengxian County, Wenxian County in Longnan City in Gansu Province, Changwu County in Shaanxi Province, Datong County in Qinghai Province, and the Ili River Valley in the Ili Kazakh Autonomous Prefecture, Xinjiang from March 2023 to May 2024, and tick species were identified morphologically using a stereomicroscope. Genomic DNA was extracted from ticks, and the 18S rRNA gene of piroplasm was amplified using nested PCR assay. Positive amplification products were sequenced, followed by sequence alignment analysis with the BLAST software. A phylogenetic tree was constructed using the neighbor-joining method with the software MEGA 11. Results A total of 1 579 ticks infesting livestock were collected and 1 559 ticks were detected, with Haemaphysalis longicornis as dominant tick species in Gansu and Shaanxi, Ixodes persulcatus in Qinghai, and Hyalomma asiaticum and Dermacentor nuttalli in Xinjiang. Nested PCR assay detected that the overall prevalence of piroplasm infection was 42.78% (667/1 559), with 47.81% (319/601) prevalence in Gansu, 30.03% (103/343) in Shaanxi, 10.71% (15/140) in Qinghai, and 48.42% (230/475) in Xinjiang. Sequencing identified six piroplasm species, including Babesia ovata, B. bigemina, B. microti, Theileria luwenshuni, T. uilenbergi, and T. orientalis, with T. orientalis, T. uilenbergi, and T. luwenshuni as dominant species in Gansu, B. microti in Qinghai, T. luwenshuni in Shaanxi, and T. orientalis, B. ovata, and B. bigemina in Xinjiang. Phylogenetic analysis showed that T. luwenshuni from Shaanxi were clustered into the same clade with the Shaanxi strain (GenBank accession number: MG930119) and Guizhou Dushan strain (GenBank accession number: KC735145), while T. orientalis from Zhangjiachuan were clustered into the same clade with the Thailand strain (GenBank accession number: MG757653) and a domestic strain (GenBank accession number: OM756747). Conclusion The prevalence of tick-borne piroplasm infection is high in northwestern China, and the dominant piroplasm species varies significantly in geographical regions.

Changes of LILRB4 expression on trophoblast cells and its impact on pregnancy caused by Toxoplasma gondii infection

ZHANG Fan, MOU Rutao, ZHANG Zhendong, LIU Xianbing, ZHANG Haixia, HU Xuemei, LI Zhidan

2026, 44(1):  85-93.  doi:10.12140/j.issn.1000-7423.2026.01.013

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Objective To investigate the changes in the expression of leukocyte immunoglobulin-like receptor subfamily B member 4 (LILRB4) on the surface of trophoblast cells following Toxoplasma gondii infection during pregnancy, and to examine its effects on trophoblast cell functions and pregnancy outcomes. Methods Human primary trophoblast cells were incubated in cell culture dishes at a density of 1 × 10⁷ cells per dish and divided into control and infected groups. Cells in the infected group were infected with T. gondii at a ratio of 1:1, incubated with APC-conjugated anti-human LILRB4 monoclonal antibody, fixed, permeabilized, incubated with APC-Cy7-conjugated anti-human cytokeratin 7 (CK7) monoclonal antibody and Alexa Fluor 488-conjugated anti-human vimentin monoclonal antibody, and the expression of LILRB4 was detected by flow cytometry. HTR-8/SVneo cells in the logarithmic growth phase were seeded into 24-well plates at a density of 2 × 10⁵ cells per well and divided into control and infected groups. Cells in the infected group were infected with T. gondii at a ratio of 1:1, incubated with mouse anti-human LILRB4 monoclonal antibody (1:200 dilution) and Elab Fluor 647-conjugated goat anti-mouse IgG monoclonal antibody (1:200 dilution) and enveloped. Images were photographed with a confocal laser scanning microscope, and the immunofluorescence intensity of LILRB4 protein was analyzed using the Image J software. Forty female and 20 male wild-type mice of the C57BL/6J strain and 40 female and 20 male LILRB4^-/- mice were randomly caged overnight, and presence of copulatory plugs in female mice on morning of the following day was considered gestational day 0. Wild-type pregnant mice were randomly assigned to wild-type control and infected groups, while LILRB4^-/- pregnant mice were randomly divided into LILRB4^-/- control and infected groups, with 6 mice in each group. Pregnant mice in wild-type and LILRB4^-/- infected groups were intraperitoneally injected with 300 T. gondii tachyzoites on day 8 of gestation, while animals in the control groups received the same volume of physiological saline. On day 14 of gestation, all mice were sacrificed and their uteri were dissected to evaluate placental and fetal development. The expression of LILRB4 protein was detected using immunohistochemistry in mouse placental tissues in wild-type control and infected groups, and the percentage of positive LILRB4 protein expression was analyzed with the software Image J. The relative expression of LILRB4 protein was determined using Western blotting in mouse placental tissues in wild-type control and infected groups. Western blotting was used to detect the relative protein expression levels of interleukin-10 (IL-10) and IL-12 in mouse placental tissues of wild-type control group, wild-type infected group, and LILRB4^-/- infected group. HTR-8/SVneo cells were seeded into cell culture dishes at a density of 1 × 10⁷ cells per dish and divided into control, infected, and LILRB4 blockade and infected group. Cells in the LILRB4 blockade and infected group were pretreated with LILRB4 neutralizing antibody for 2 hours, while cells in the infected and LILRB4 blockade and infected groups were infected with T. gondii at a ratio of 1:1. The relative expression of IL-10 and IL-12 proteins was quantified using Western blotting in HTR-8/SVneo cells in control, infected and LILRB4 blockade and infected groups. The invasive ability of HTR-8/SVneo cells was evaluated using Transwell assay following T. gondii infected. All statistical analyses were performed using the software GraphPad Prism 9.0. Difference of means between groups was tested for statistical significance with independent-sample t-test, and multiple-group comparisons were conducted with one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test. Results Flow cytometry detected a higher percentage of LILRB4-positive human primary trophoblast cells in the control group than in the infected group [(26.10 ± 1.99)% vs. (18.60 ± 1.13)%; t = 15.00, P < 0.01], and immunofluorescence staining revealed that the mean fluorescence intensity of LILRB4 protein was lower in the infected group than in the control group [(122.56 ± 5.24) vs. (149.27 ± 3.50); t = 5.36, P < 0.05]. Pregnant mice in the wild-type and LILRB4^-/- control groups presented shiny fur, normal mental state, and well placental and fetal development, while pregnant mice in the wild-type and LILRB4^-/- infected groups exhibited remarkable low spirits, shaggy and rough fur, placental ischemia, and poor fetal growth. The mouse placental [(54.82 ± 7.12) mg vs. (72.51 ± 1.11) mg; t = 4.25, P < 0.05] and fetal weights [(140.59 ± 3.19) mg vs. (201.03 ± 17.37) mg; t = 5.92, P < 0.01] were lower in the wild-type infected group than in the wild-type control group, and the mouse placental [(41.24 ± 2.80) mg vs. (54.82 ± 7.12) mg; t = 3.07, P < 0.05] and fetal weights [(68.25 ± 11.55) mg vs. (140.59 ± 3.19) mg; t = 10.45, P < 0.01] were lower in the LILRB4^-/- infected group than in the wild-type infected group. Immunohistochemical staining showed that the LILRB4 protein was highly positive in mouse placental tissues in the wild-type control group, with positive LILRB4 protein expression primarily found in cell membranes, and low LILRB4 protein expression was found in the wild-type infected group. The proportion of positive LILRB4 protein expression was lower in the wild-type infected group than in the wild-type control group [(16.13 ± 2.55)% vs. (36.64 ± 6.62)%; t = 5.00, P < 0.01]. Western blotting showed that the relative protein expression of LILRB4 in mouse placental tissues was higher in the wild-type control group than in the wild-type infected group [(1.15 ± 0.05) vs. (0.78 ± 0.10); t = 5.40, P < 0.05], and the relative IL-10 protein expression was lower in mouse placental tissues in the wild-type infected group than in the wild-type control group [(0.93 ± 0.09) vs. (1.28 ± 0.16); Tukey’s post hoc test, P < 0.05], and lower in the LILRB4^-/- infected group than in the wild-type infected group [(0.61 ± 0.10) vs. (0.93 ± 0.09); Tukey’s post hoc test, P < 0.05]. The relative IL-12 protein expression was higher in mouse placental tissues in the wild-type infected group than in the wild-type control group [(1.08 ± 0.11) vs. (0.55 ± 0.18); Tukey’s post hoc test, P < 0.05], and higher relative IL-12 protein expression was detected in the LILRB4^-/- infected group than in the wild-type infected group [(1.67 ± 0.29) vs. (1.08 ± 0.11); Tukey’s post hoc test, P < 0.05]. The relative IL-10 protein expression was lower in HTR-8/SVneo cells in the infected group than in the control group [(0.85 ± 0.05) vs. (1.15 ± 0.06); Tukey’s post hoc test, P < 0.05], and lower in the LILRB4 blockade and infected group (0.72 ± 0.04) than in the infected group (Tukey’s post hoc test, P < 0.05), and the relative IL-12 protein expression was higher in HTR-8/SVneo cells in the infected group than in the control group [(0.89 ± 0.10) vs. (0.52 ± 0.14); Tukey’s post hoc test, P < 0.05], and higher in the LILRB4 blockade and infected group than in the infected group [(1.21 ± 0.04) vs. (0.89 ± 0.10); Tukey’s post hoc test, P < 0.05]. In addition, the counts of invasive HTR-8/SVneo cells were lower in the infected group than in the control group [(178 ± 21) vs. (278 ± 18); t = 45.60, P < 0.01], and lower in the LILRB4 blockade and infected group (119 ± 9) than in the infected group (t = 5.50, P < 0.05). Conclusion T. gondii infection may significantly downregulate LILRB4 protein expression in human trophoblast cells and mouse placental tissues, and downregulation of LILRB4 promotes IL-12 expression and inhibits IL-10 production, thereby attenuating the invasive ability of trophoblast cells.

Species composition, host specificity and genetic diversity of ectoparasites on rodents in Shiqu County, Sichuan Province

YANG Yang, WANG Xu, LI Mengqing, XUE Chuizhao, ZUO Qingqiu, YIN Jianhai, CAO Jianping

2026, 44(1):  94-101.  doi:10.12140/j.issn.1000-7423.2026.01.014

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Objective To investigate the species composition, host associations, and genetic structure of ectoparasites on rodents from alpine meadow habitats in Shiqu County, Sichuan Province, so as provide baseline data for risk assessment of regional zoonoses. Methods Small rodents were captured using random quadrats with traps in Shiqu County, Sichuan Province in October 2023 and August 2024, and identified morphologically. Ectoparasites were collected from the host rodents by fur-combing, and DNA was extracted. Flea 18S rRNA, mite cytochrome c oxidase subunit 1 (cox1) and tick 16S rDNA gene fragments were amplified using PCR assay for species identification. The obtained sequences were subjected to homology analysis. Haplotypes were analyzed using the software DnaSP 6.12.03, and phylogenetic trees were constructed using the maximum likelihood method in MEGA 7.0 to infer genetic evolutionary characteristics. In addition, a quantitative host-parasite association network was visualized as a Sankey diagram using the software Python 3.13.2. Results A total of 472 small rodents belonging to 7 species were captured, with Neodon fuscus (52.12%, 246/472) and Ochotona curzoniae (35.81%, 169/472) as dominant species. A total of 147 ectoparasites were collected and classified into 32 species, including 13 fleas species (94 individuals), 6 tick species (11 individuals), and 13 mite species (42 individuals), with Oropsylla silantiewi and Foxella ignota as the most abundant flea species (both accounting for 24.47%, 23/94), Poecilochirus austroasiaticus as the most abundant mite species (40.48%, 17/42), and Dermacentor silvarum and D. andersoni as the most abundant tick species (both accounting for 3/11). Fleas, mites and ticks displayed 30, 28, and 10 haplotypes, with haplotype diversity values of 0.863 5, 0.981 8 and 1.000 0, respectively, indicating significant genetic differentiation. Phylogenetic analysis showed clear clustering of distinct taxa. O. curzoniae and N. fuscus harbored 23 and 16 parasite species, respectively, representing core host nodes. 87.5% (28/32) of parasite species exhibited host specificity, with 65.6% (21/32) infesting only single host species. Conclusion Rodents harbor a rich diversity of ectoparasites in Shiqu County, Sichuan Province, with fleas, ticks, and mites as the main parasite groups, and O. curzoniae and N. fuscus as key hosts. The ectoparasite community exhibits a high host specificity, considerable haplotype richness, and pronounced genetic diversity, with phylogenetic analysis revealing distinct clustering patterns.

From concept to practice: dilemmas and mechanism innovations in the implementation of One Health at the grassroots level in China

XIE Hongli, LU Danping, GAO Yang

2026, 44(1):  102-109.  doi:10.12140/j.issn.1000-7423.2026.01.015

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Objective To investigate the dilemma and optimization path for implementation of the One Health concept at the grassroots level in China, so as to provide the decision-making evidence for grassroots health governance in China. Methods A total of 30 workers were sampled from grassroots government departments related to One Health in Beijing, Zhejiang, Hainan, and Qinghai from June to August in 2023, including provincial-, municipal-, and county-level health commissions, centers for disease control and prevention, departments of ecology and environment, agriculture departments, transportation departments, and administrations for market regulation) as interviewees. Semi-structured interviews were conducted using the convenience sampling method to collect practical experiences, current situations, and problems related to the multi-sectoral health collaboration of the One Health concept. The interview data were subjected to open coding, axial coding, selective coding and analysis using the software NVivo 12. Results Following open coding of interview data， a total of 51 initial concepts, along with 25 sub-categories were yielded, including preparation and response, safety risk, information exchange, concept acceptance, management and coordination, communication methods, social benefits, collaboration mechanisms, collaborative governance, cross-regional collaboration, policy and planning, incentive mechanisms, international cooperation, audit plans, departmental responsibilities, joint planning, cross-sectoral collaboration, information sharing, economic conditions, health conditions, publicity, resources, evaluation mechanisms, feedback mechanisms, and supervision and implementation. Following axial coding and re-clustering, these 25 sub-categories were condensed into 10 main categories, namely information exchange, policy and planning, incentive mechanisms, social benefits, dissemination, collaboration mechanisms, communication, resources, preparation and response, and international cooperation. Axial coding and selective coding revealed that multisectoral collaboration was notably crucial in implementation of the One Health concept, and this collaboration was embodied in cross-sectoral cooperation, emergency response, data sharing, social responsibility, and global collaboration. Conclusion Institutional innovation is critical to implementation of One Health, which requires the establishment of grassroots mechanisms to achieve health coordinated governance of “human-animal-environment”. It is necessary to implement the strategy of “preventing human diseases by controlling animal diseases, and implementation of interventions moving forward from the source”, and strengthen the prevention at the source. Future efforts should focus on expansion of pilot programs, popularization of assessment tools, and extension of policies to communities.

STANDARD INTERPRETATION

Interpretation of the standard Detection of Plasmodium spp. — Nucleic Acid Identification of the Species

LI Mei, YAN He, XIA Zhigui, ZHENG Bin, YU Chenghang, XIAO Ning

2026, 44(1):  110-117.  doi:10.12140/j.issn.1000-7423.2026.01.016

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The standard Detection of Plasmodium spp. — Nucleic Acid Identification of the Species was formulated according to regulations defined in Guidelines for Health Standard Management, and GB/T 1.1-2020 Directives for Standardization - Part 1: Rules for the structure and drafting of standardizing documents. The standard includes seven sections, which are scope of application, normative reference documents, terms and definitions, reagents and materials, instruments and equipment, samples, and detection procedures, and two informative appendices and one normative appendix. This standard has been issued by the National Disease and Prevention Administration (WS/T 10030-2025) and will be effective since March 1, 2026. The implementation of this standard will provide a unified technical document and operating guidelines for nucleic acid identification of Plasmodium species across centers for disease control and prevention and healthcare institutions at all levels in China, which will further improve the reliability, practicality, and operability of nucleic acid dentification of malaria parasites.

REVIEWS

Progress researches on Budd-Chiari syndrome and portal hypertension caused by hepatic alveolar echinococcosis

A Jide, CHAI Jinping, JING Qindong, PAN Hongshuai

2026, 44(1):  118-123.  doi:10.12140/j.issn.1000-7423.2026.01.017

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Hepatic alveolar echinococcosis is a zoonotic parasitic disease that is known as the parasite cancer. Lesions of hepatic alveolar echinococcosis tend to grow along the intrahepatic blood vessels, and large lesions may directly invade or compress adjacent portal veins, hepatic vein, trunk, and the post-hepatic segment of the inferior vena cavar esulting in that the majority of patients with end-stage hepatic alveolar echinococcosis are complicated with cardiac cirrhosis, Budd-Chiari syndrome, or portal hypertension at varying degrees. This not only increases the difficulty in diagnosis and treatment for hepatic alveolar echinococcosis, but also raises the cost of disease treatment. Currently, the pathogenesis of hepatic alveolar echinococcosis induced portal hypertension, Budd-Chiari syndrome, and cardiac cirrhosis is not fully understood, and there are still controversies regarding diagnosis and treatment of these complications. This article provides a brief overview of the clinical symptoms induced by hepatic alveolar echinococcosis, including cardiac cirrhosis, Budd-Chiari syndrome, and portal hypertension.

Advances in the effects of parasitic protozoan infections on the host gut microbiota

LI Pengyao, LI Jing, ZHENG Bin, LU Shaohong

2026, 44(1):  124-129.  doi:10.12140/j.issn.1000-7423.2026.01.018

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Gut microbiota, as a crucial regulator of host health, has received growing attention for its role in parasitic protozoan infections. A variety of parasitic protozoa may induce host gut microbiota dysbiosis through direct invasion of intestinal epithelium, activation of inflammatory responses, disruption of intestinal barriers, or alterations in metabolic pathways, and this dysbiosis is characterized by a decrease in beneficial bacteria, proliferation of pathogenic bacteria, and a reduction in microbial diversity. Toxoplasma gondii and Giardia lamblia have been found to induce microbiota remodeling and persistently affect host metabolism, while the development of Leishmania and Trypanosoma in the intestines of their transmitting vectors are remarkably affected by microbiota. In addition, symbiotic bacteria may promote or inhibit pathogen growth and differentiation. Gut microbiota not only affects parasite colonization and infection severity but also modulates host immune responses through metabolites such as short-chain fatty acids, indoles, and bile acids, which is a non-negligible key regulator during parasitic infections. This review summarizes the alterations in gut microbiota caused by major parasitic protozoan infections and their potential mechanisms of actions, so as to provide insights into better understanding of pathogenesis and development of microecological intervention strategies.

Progress of researches on biological therapies targeting type 2 inflammation and risk of parasitic infections

CAI Wenlin, LIU Xu, HU Ke

2026, 44(1):  130-136.  doi:10.12140/j.issn.1000-7423.2026.01.019

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Recently, biologics targeting type 2 inflammatory pathways, such as interleukin-4 (IL-4)/IL-13, IL-5, and immunoglobulin E (IgE), have become effective approaches for treatment of type 2 inflammatory diseases. Nevertheless, these biologics not only precisely suppress excessive inflammatory responses, but also interfere with the body’s capability to fight against parasitic infections, thereby increasing the risk of infection. Although such adverse events are relatively rare, their potential associations have been paid attention in clinical practices. Currently, there is still a lack of systematic understanding regarding the exact mechanisms underlying parasitic infections induced by biologics with diverse targets, the characteristics of high-risk populations, and clinical management strategies. This article aims to provide a review of these aspects, so as to provide insights into clinical practices.

SHORT COMMUNICATIONS

Impact of Demodex infestation on the physiological functions of human facial skin

LIU Siwen, ZHANG Dacun, LI Zhiqiang, LIU Chunsheng, CHEN Si, TIAN Ruyu, HUANG Zeyu, SONG Ziyue, QIN Lei, ZHANG Ruzhi, LIU Xiaolv, GU Shengli, ZHAN Xiaodong

2026, 44(1):  137-140.  doi:10.12140/j.issn.1000-7423.2026.01.020

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To investigate the impact of Demodex mite infestation on the physiological functions of human facial skin, 240 university students were screened for Demodex infections using the transparent adhesive tape method. Base on the inclusion criteria, 32 students were tested positive for Demodex. Skin physiological function tests (skin erythema, melanin content, moisture, sebum, pore size, elasticity, and gloss) and skin image analysis were conducted among subjects positive for Demodex prior to treatment. Following testing, patients were given oral metronidazole tablets at a single dose of 0.4 g, three times daily, for successive 7 days. Facial skin was tested for presence of mites with transparent adhesive tapes 2 weeks post-treatment. Then, non-affected areas in the left-side mite-free face were randomly sampled for skin physiological function tests and skin image analysis, and the skin physiological and imaging parameters on face were compared between pre- and post-treatment. The facial skin erythema value, sebum secretion, proportion of skin pores, and gloss were 16.42 ± 16.48, (18.59 ± 3.58) µg/cm², (4.13 ± 0.31)%, and (3.59 ± 0.22) GU, respectively among patients tested positive for Demodex post-treatment, which were significantly lower than those [20.99 ± 17.79, (22.95 ± 4.42) µg/cm², (4.61 ± 0.33)%, (4.07 ± 0.23) GU] pre-treatment (t = 2.26, 3.86, 3.24, 2.29; all P < 0.05). The skin water content higher post-treatment than pre-treatment [(51.06 ± 11.41) a.u. vs. (39.18 ± 9.30) a.u.; t = -5.31, P < 0.01], and the melanin content (151.74 ± 6.80 vs. 155.56 ± 6.50; t = 1.14, P > 0.05) and gross elasticity (R₂) (63.04 ± 0.98 vs. 64.77 ± 1.51; t = 1.09, P > 0.05) were numerically lower post-treatment than pre-treatment. Skin image analysis revealed that, on near-infrared images, patients positive for Demodex exhibited facial erythema and extensive capillary dilation prior to treatment, and reduced erythematous areas and remarkable alleviation of capillary dilation and inflammatory acne post-treatment. On red channel images, extensive deep red areas were found on facial skin among patients positive for Demodex prior to treatment, and diminished deep red zones and partially turned pink were observed post-treatment, with markedly reduced deep inflammatory areas. These findings suggest that Demodex infestation may affect skin physiological functions, and lead to imbalance of water and sebum secretion in skin, thereby impairing the skin barrier function, and promoting capillary dilation, and increasing hemoglobin levels.

Analysis on 21 cases of primary echinococcosis in the pleural cavity

JING Xiaoliang, ZHANG Liwei

2026, 44(1):  141-143.  doi:10.12140/j.issn.1000-7423.2026.01.021

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The medical records of 21 patients with primary echinococcosis in the pleural cavity that received surgical treatment in the First Affiliated Hospital of Xinjiang Medical University from January 2000 to April 2025 were retrospectively collected, and a descriptive epidemiological method was employed to analyze the demographic features, epidemiological characteristics, clinical manifestations, chest CT scans, treatment and outcomes. The case series included 14 men and 7 women and had a mean age of 44.8 years. There were 11 cases with a history of staying in epidemic areas or contact with canines. There were 15 cases with echinococcosis in the right-side pleural cavity, 14 cases identified using physical examinations, 15 cases with multiple hydatid cysts, and 19 cases with a medical history of hepatic echinococcosis. Surgical interventions consisted of thoracoscopic surgery among 6 cases and thoracotomy among 16 cases. Postoperative complications included pulmonary infection (8 cases), pneumothorax (2 cases), and empyema (1 case), and conventional postoperative pathological examinations revealed cystic echinococcosis in 16 cases and alveolar echinococcosis in 5 cases. Of all cases, there were 4 cases lost to the 36-month follow-up after discharge from hospital, and among the 17 cases completing follow-up, there were 2 cases with recurrence 12 and 15 months post-surgery and 15 cases without recurrence. These findings demonstrate that preoperative chest CT scan is the optimal choice for evaluation of parasitic diseases primarily originating from the pleural cavity, and thoracoscopic resection may represent a feasible and safe therapeutic strategy for selected patients following thorough professional assessment.

CASE REPORTS

Imported ocular loaiasis in Jilin Province: a case report

LU Yan, CUI Jun, ZHOU Fuxian, CUI Renzhe, TIAN Lianji

2026, 44(1):  144-147.  doi:10.12140/j.issn.1000-7423.2026.01.022

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The patient was a 44-year-old, self-employed, Han ethnic man residing in Dunhua City, Jilin Province. The patient had resided in Gabon, Africa for 5 years due to labor and returned to China in June 2025. On August 9, 2025, the patient visited Yanbian University Hospital with complaints of a foreign body sensation and crawling sensation in the right eye for one day. Routine blood tests showed an eosinophil percentage of 34.7% and an eosinophil count of 3.73 × 10⁹/L. Ophthalmic slit-lamp examinations identified transparent filamentous worms under the conjunctiva of the right eye, exhibiting a wriggling motion, and right eye conjunctival congestion without edema. At approximately 12: 00 on the same day, a subconjunctival parasite extraction was performed under local anesthesia, and the worm, located in the temporal side of the right eye conjunctiva and curled in shape, was completely removed, and identified by the Department Laboratory Medicine of Yanbian University Hospital and Department of Parasitology of Yanbian University Medical College. The worm surface was covered with irregular, smooth, circular, semi-transparent warty protrusions, with sheathed microfilariae in the uterus, and the parasite morphology was consistent with the description of female Loa loa in the literature. The case was diagnosed as imported loaiasis, and postoperatively given antibiotic eye ointment in the conjunctival sac, tobramycin/dexamethasone eye drops and levofloxacin eye drops (one drop each, 4 times daily). Peripheral blood was sampled postoperatively, and no microfilariae were detected in the blood smears by microscopy. A follow-up examination one week post-surgery revealed the patient had self-administered ivermectin tablets (a single oral dose of 150 μg/kg, 2-week treatment course) according to the mass deworming protocol in Gabon. On September 29, 2025, the patient voluntarily visited Beijing Friendship Hospital for a follow-up examination, where L. loa antigen tests yielded a negative result. Albendazole tablets (0.4 g/d) were additionally given, and regular monitoring of blood counts and liver/kidney function was initiated.

A case of migratory pneumonia caused by Trichuris trichiura infection

MO Jun, WU Wei, CHEN Ying

2026, 44(1):  148-150.  doi:10.12140/j.issn.1000-7423.2026.01.023

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A 13-year-old male patient was admitted to the Outpatient Department of Jiangsu Institute of Parasitic Diseases on July 1, 2025, due to decreased appetite with no weight gain for more than half a year. Repeated CT scans showed pulmonary inflammation and migratory lesions. Routine blood tests showed an increased eosinophil count and percentage, and serological tests indicated positive antibodies against multiple parasites. Modified Kato-Katz thick smear technique of stool samples displayed spindle-shaped Trichuris trichiura eggs under a microscope. The case resided in Bijie, Yunnan Province, and had a history of consuming raw water and food. Migratory pneumonia caused by T. trichiura infection was diagnosed based on the patient’s epidemiological history, clinical manifestations and laboratory findings. The case was given albendazole by oral gavage (a single dose of 200 mg, twice daily for 3 consecutive days). Blood routine tests one week post-treatment showed a decrease in eosinophils, and chest CT scans displayed alleviated pulmonary inflammation, with no parasite eggs identified in stool samples. The patient’s appetite improved remarkably with no other discomfort 2 weeks post-treatment.

A rare case report of ectopic parasitism: Mammomonogamus in human jejunum

XU Ruina, YE Zuowan, WANG Jing, MIAO Feng

2026, 44(1):  151-152.  doi:10.12140/j.issn.1000-7423.2026.01.024

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The patient was a 72-year-old male from Nanyang, Henan Province, with a permanent residence in Shenzhen City, who had a habit of fishing using earthworms as baits. He underwent a gastrectomy for gastric cancer in 2023, and nematodes were found parasitizing the jejunum during a routine electronic gastroscopic follow-up; however, but no typical clinical manifestations of respiratory or digestive system were observed. Routine blood tests showed a white blood cell count of 6.61 × 10⁹/L, an eosinophil percentage of 0.7%, and an absolute eosinophil count of 0.05 × 10⁹/L, with all indicators within the normal range. The nematodes were removed using biopsy forceps, appearing red, slender, and atypical in shape with a “Y” configuration, measuring approximately 10 mm in length and capable of autonomous movement. Following staining with Harris’s hematoxylin, the nematodes were identified as male Mammomonogamus under an optical microscope. The patient was diagnosed with Mammomonogamus infection and orally administered with albendazole (400 mg/d, once daily for one week). No Mammomonogamus eggs were detected in the patient’s stool samples one week post-treatment.