Table of Content

30 December 2025, Volume 43 Issue 6

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SPECIAL REPORTS

Progress of echinococcosis control in China, 2024

LIU Baixue, KUI Yan, XUE Chuizhao, WANG Xu, WANG Ying, WANG Liying, YANG Shijie, HAN Shuai, LI Shizhu

2025, 43(6):  739-748.  doi:10.12140/j.issn.1000-7423.2025.06.001

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Objective To investigate the progress of nationwide echinococcosis control program in China in 2024, and to summarize the experiences and challenges, so as to provide insights into optimization of echinococcosis control strategies and measures. Methdos Data pertaining to national echinococcosis control programs in endemic areas of China in 2024 were collected from the Information Management System for Parasitic Disease Control operated by Chinese Center for Disease Control and Prevention to create a database. A descriptive analysis was performed to understand human echinococcosis tests and treatments, prevalence of infections in infectious sources, and prevalence of echinococcosis in intermediate hosts, and compare them with the data from 2020 to 2023. Intergroup comparisons were performed using Pearson’s chi-square test and Cochran-Armitage trend test. Results A total of 23 622 prevalent echinococcosis cases were found in 370 endemic counties across China by the end of 2024, with a prevalence rate of 53.73/10⁵ (23 622/43 963 728), appearing a 15.40% decrease relative to 2020 (63.51/10⁵). A total of 2 159 incident echinococcosis cases were reported in China in 2024, showing a 13.63% increase relative to 2020 (1 900 cases), and 10.79% (233/2 159) of cases were detected among individuals at ages of under 12 years. Abdominal ultrasound scans were performed for human echinococcosis screening in China at 5.640 4 million person-times in 2024, and suspected cases positive for ultrasound scans were subjected to serological tests at 12 479 person-times. A total of 16 717 individuals received chemotherapy against echinococcosis and 2 061 individuals received surgical treatments for echinococcosis in 2024, including 69.29% of cystic echinococcosis patients (1 428/2 061) and 24.55% of alveolar echinococcosis patients (506/2 061). The 2024 follow-up outcomes revealed that there were 2 499 cases with cure, 18 294 cases with responses to treatments, 4 511 cases with failure in treatments, 371 deaths (the causes of the deaths were not echinococcosis), 483 cases excluded from the study, 231 cases lost to follow-up, 169 cases with incompletion of the follow-up, and 88 cases migrating to other places. In 2024, there were 2 073 297 dogs in echinococcosis-endemic townships in China, including 1 960 195 dogs registered for management. Deworming was conducted in dogs in 35 146 villages, including domestic dog deworming at 20 989 364 times, and wild canines deworming with 347 167 doses. The prevalence of positive Echinococcus coproantigen results was 0.51% (2 109/411 144) in domestic dogs, appearing a tendency towards a rise since 2021 (Z = 3.66, P < 0.05), and was 3.33% (2 437/73 088) in wild canines, showing a tendency towards a rise since 2021 (Z = 31.37, P < 0.05). In 2024, the prevalence of echinococcosis was 0.92% (1 273/138 373) in livestock, appearing a tendency towards a rise since 2021 (Z = 10.20, P < 0.05), and was 0.57% (282/49 780) in field rodents, appearing a tendency towards a decline since 2021 (Z = -11.31, P < 0.05). Conclusion The prevalence of echinococcosis had been basically controlled in China in 2024; however, the rebound of incident cases and an increase in the prevalence of echinococcosis in infectious sources indicate a risk of resurgence. Future echinococcosis control efforts should continue to strengthen integrated control and cross-sectoral collaboration, and focus on precise management of infectious sources in the context of the One Health concept, improved grassroots capacity, and increased investment in scientific research to overcome key technical bottlenecks, so as to improve echinococcosis control capabilities in China.

Epidemiological characteristics of visceral leishmaniasis in China in 2024

LI Yuanyuan, ZHOU Zhengbin, LI Zhongqiu, YANG Limin, LIU Qin, ZHANG Yi, CHEN Junhu, LI Shizhu

2025, 43(6):  749-756.  doi:10.12140/j.issn.1000-7423.2025.06.002

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Objective To investigate the epidemiological characteristics of visceral leishmaniasis in China in 2024， so as to provide insights into formulation of visceral leishmaniasis control measures and strategies. Methods Epidemiological data of visceral leishmaniasis cases in China in 2024 were collected from National Notifiable Diseases Reporting Information System operated by Chinese Center for Disease Control and Prevention. After excluding suspected cases, duplicates and cutaneous leishmaniasis cases, all data were input into Microsoft Excel 2016, and the epidemiological characteristics of visceral leishmaniasis cases were analyzed using at descriptive epidemiological method. Results A total of 278 visceral leishmaniasis cases were reported in 120 counties across 14 provinces (autonomous regions and municipalities) in China in 2024, with a decrease of 7.02% in relative to in 2023 (299 cases), and including 232 local cases reported in 77 endemic counties, and 46 imported cases in non­endemic counties. There were 225 cases with anthropozoonotic (mountain-type zoonotic) visceral leishmaniasis (80.93%), 5 cases with euzoonotic (desert-type zoonotic) visceral leishmaniasis, and 2 cases with anthroponotic visceral leishmaniasis, and these reported cases were predominantly distributed in three provinces of Shanxi (108 cases), Henan (53 cases) and Shaanxi (32 cases), accounting for 69.42% (193/278) of total cases in China. In 77 endemic counties, Pingding County (18 cases), Suburb District (11 cases) of Yangquan City in Shanxi Province, Linzhou City (13 cases) and Dengfeng City (11 cases) of Henan Province and Jingxing County (12 cases) of Hebei Province had relatively high counts of incident cases, accounting for 23.38% (65/278) of total cases in China. The counties with recurrent visceral leishmaniasis were mainly concentrated in Shanxi Province (Yuci District, Zuoquan County, Taigu District, Zezhou District, Wenxi county and Shangdang District), Henan Province (Shaanzhou District, Lushi County, Yindu District, Yiyang County and Ruzhou City), Hebei Province (Yuanshi County, Xiongxian County and Xiahuayuan District), Shaanxi Province (Chengcheng County and Mizhi County) and Beijing City (Fangshan District), with a total of 26 local cases reported. The incidence of visceral leishmaniasis peeked in August, and there were 191 male cases and 87 femles cases, respectively. The reported visceral leishmaniasis cases were found at ages of 45 to 74 years, and farmers were the predominant occupation (53.96%, 150/278). Conclusion The prevalence of visceral leishmaniasis is low in China; however, the endemic areas of mountain-type zoonotic visceral leishmaniasis is gradually expanding. Intensified surveillance and management of mountain-type zoonotic visceral leishmaniasis is recommended.

ORIGINAL ARTICLES

Spatio-temporal distribution characteristics of alveolar echinococcosis in Xiji County, Ningxia Hui Autonomous Region, 2005-2024

XIAN Yuying, TIAN Huifang, FU Weicheng, YAN Fang, XUE Chuizhao, WANG Ying, WU Xianglin, WANG Xu

2025, 43(6):  757-765.  doi:10.12140/j.issn.1000-7423.2025.06.003

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Objective To comprehensively analyze the spatiotemporal distribution of human alveolar echinococcosis (AE) in Xiji County and to identify underlying environmental factors, thereby providing a scientific basis for targeted prevention and control strategies. Methdos Surveillance data on human AE case in Xiji County (2005-2024) were collected. The annual number of cases by township was visualized using heatmaps. Joinpoint regression models were utilized to analyze temporal trends in incidence rates, calculating the average annual percentage change (AAPC) and annual percentage change (APC). R language packages were used to compute and plot the trends in annual prevalence rates. ArcGIS was employed for spatial visualization, global and local Moran’s I index analyses to assess spatial autocorrelation, and SaTScan was performed for space-time scan analysis to identify spatiotemporal clusters. A multivariable logistic regression model were constructed to analyze the association between plant formation area and AE occurrence at the village level. Results A total of 325 human AE cases were identified in Xiji County over the 20-year period, with the peak incidence rate occurring in 2007 (33.87/100 000). The incidence rate has shown a significant downward trend (AAPC = -13.27%, P < 0.01) in the past 15 years, and the prevalence rate of AE also showed a consistent decline (Z = -21.98, P < 0.01) from 2015 to 2024. The average annual incidence (countywide: 4.03/100 000) showed an uneven geographical distribution: higher in northern townships (Xinying: 31.64/100 000; Baiya: 30.63/100 000) and lower in the south. Spatiotemporal scan analysis identified a significant primary cluster encompassing Xinying, Baiya, and Huoshizhai townships for the period 2007-2009 (RR = 30.90, LLR = 288.76, P < 0.01). Cases were reported in 34.09% of all villages/communities, with exceptionally high endemic village proportions within Xinying (95.45%) and Baiya (92.86%) townships. Logistic regression revealed that increased area coverage of Stipa bungeana steppe, cropland, and Artemisia giraldii-grass steppe significantly elevated the AE risk at the village level (OR = 1.32, 1.12, 1.44; all P < 0.05). Conclusion The AE epidemic in Xiji County is characterized by a historically high burden in the north, significant spatiotemporal clustering, and a marked overall decline over the past 15 years. Key vegetation biomes are significant risk factors, elevating AE risk by shaping suitable habitats for rodent intermediate hosts and influencing human exposure activities. Although comprehensive control strategies have significantly reduced the AE incidence and prevalence rates in Xiji County, persistent transmission risks remain.

Effect of early Echinococcus multilocularis infection on natural killer T cell subsets and their function in murine livers

TAN Wei, AYINUER Aierken, REXIATI Ruze, YUAN Zhongdian, ZHU Dalong, YANG Chengming, TUERGANAILI Aji

2025, 43(6):  765-776.  doi:10.12140/j.issn.1000-7423.2025.06.004

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Objective To examine the effects of early Echinococcus multilocularis infection on the quantity, subsets, and functions of natural killer T (NKT) cells in mouse hepatic tissues. Methdos C57BL/6N mice were randomly divided into a control group and an infection group. Mice in the infection group were inoculated with 2 000 E. multilocularis protoscoleces via the hepatic portal vein, while animals in the control group received an equal volume of physiological saline. Mouse liver tissues were collected 4 weeks post-infection. The pathological changes of liver tissues were observed by hematoxylin-eosin (HE) staining, and alterations in hepatic NKT cells were examined using immunofluorescence staining. Lymphocyte suspensions were prepared from mouse liver tissues. The percentages of NKT cells and their different subsets were quantified in mouse liver tissues in both control and infection groups using flow cytometry, and the expression of inhibitory receptors [natural killer group 2 member A （NKG2A） and programmed cell death protein 1 (PD-1)], an activating receptor NKG2D, chemokine receptors [C-X-C motif chemokine receptor 3 (CXCR3) and CXCR6], interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), and transforming growth factor-β1 (TGF-β1) was detected in NKT cells and their respective subsets. Results HE staining revealed lesions exhibiting a germinal layer structure in mouse liver tissues in the infection group 4 weeks post-infection with E. multilocularis, surrounded by large number of inflammatory cells. Immunofluorescence staining showed that the average count of NKT cells was (1.20 ± 0.53) cells per field in the liver tissues of the infected group, which was significantly lower than that in the control group (3.73 ± 0.50) cells (t = 6.01, P < 0.01). Flow cytometry detected that the proportions of NKT cells and CD69⁺NKT cells were (11.83 ± 1.19)% and (76.93 ± 3.06)% in mouse liver tissues in the infection group, which were lower than those [(23.20 ± 0.20)% and (88.47 ± 1.53)%, respectively] in the control group (t = 16.28 and 5.84; both P < 0.01). The proportions of hepatic NKT cells secreting IFN-γ and IL-10 were (17.50 ± 5.04)% and (4.54 ± 0.20)% in the infection group, which were lower than those [(61.90 ± 2.25)% and (10.24 ± 1.37)%, respectively] in the control group (t = 13.93, 7.15, P < 0.01, 0.05). The proportion of hepatic NKT cells secreting TGF-β1 was higher in the infected group [9.04% (8.01%, 9.62%)] than in the control group [3.68% (3.48%, 3.68%)] (t = 8.60, P < 0.05), and there was no significant difference in the proportion of hepatic NKT cells producing TNF-α between the infection group [(20.83 ± 5.75)%] and the control group [(19.20 ± 5.91)%] (t = 0.34, P > 0.05). The proportions of NKG2A⁺NKT [(13.90 ± 3.18)% vs. (22.37 ± 2.61)%; t = 3.57, P < 0.05], CXCR3⁺NKT [(26.77 ± 6.00)% vs. (55.40 ± 13.64)%; t = 3.33, P < 0.05] and CXCR6⁺NKT cells [(57.20 ± 7.63)% vs. (86.73 ± 2.72)%; t = 6.31, P < 0.01] were significantly lower in mouse liver tissues in the infection group than in the control group. The percentage of PD-1⁺NKT cells [(10.04 ± 1.01)% vs. (6.56 ± 0.10)%; t = 5.92, P < 0.05] was significantly higher in mouse liver tissues in the infection group than in the control group, and there was no significant difference in the proportion of NKG2D⁺NKT cells in mouse liver tissues between the infection group and the control group [1.58% (1.56%, 2.14%) vs. 1.25% (1.15%, 1.31%); Z = ‒1.96, P > 0.05]. The proportions of CD4⁺NKT cells [(42.67 ± 4.15)% vs. (60.97 ± 1.80)%; t = 7.01, P < 0.01] and CD69⁺CD4⁺NKT cells [(44.80 ± 2.27)% vs. (62.30 ± 3.16)%; t = 7.79, PP < 0.01] were significantly lower in mouse liver tissues in the infection group than in the control group, and the proportions of CD4⁻CD8⁻ (double-negative, DN) NKT cells [(45.63 ± 3.61)% vs. (32.57 ± 2.06)%; t = 5.44, P < 0.01] and CD69⁺DN NKT cells [(44.53 ± 2.23)% vs. (32.17 ± 1.34)%; t = 8.23, P < 0.01] were higher in mouse liver tissues in the infection group than in the control group. The proportions of CD4⁺NKT cells secreting IL-10 [(9.97 ± 4.38)% vs. (52.00 ± 4.20)%; t = 11.99, P < 0.01], and DN NKT cells producing IFN-γ [(15.87 ± 2.57)% vs. (59.80 ± 4.70)%; t = 14.20, P < 0.01] and TNF-α [(6.61 ± 2.69)% vs. (30.57 ± 2.02)%; t = 12.33, P < 0.01] were lower in mouse hepatic tissues in the infection group than in the control group, and the proportions of CD8⁺NKT cells secreting IL-10 [(55.90 ± 13.68)% vs. (10.10 ± 3.24)%; t = 5.64, P < 0.01] and TGF-β1 [(45.87 ± 5.95)% vs. (19.69 ± 10.25)%; t = 3.83, P < 0.05] were significantly higher in mouse liver tissues in the infection group than in the control group. The proportions of NKG2A⁺CD4⁺NKT cells [(37.87 ± 4.80)% vs. (52.80 ± 5.17)%; t = 3.66, P < 0.05] and CXCR3⁺CD4⁺NKT cells [(40.40 ± 9.48)% vs. (57.00 ± 3.90)%; t = 2.81, P < 0.05] were significantly lower in the infection group than in the control group, and the proportion of NKG2A⁺DN NKT cells was higher in the infection group [(55.63 ± 3.46)%] than in the control group [(41.43 ± 2.31)%] (t = 5.91, P < 0.01). Conclusion During the early stage of E. multilocularis infection, NKT cells exhibite an overall state of functional suppression or exhaustion in mouse liver tissues. The NKT cell subsets demonstrate distinct expression characteristics: CD4⁺NKT cells show low expression, and CD8⁺NKT cells function by secreting IL-10 and TGF-β1, while the highly activated DN NKT cells have an impaired cytokine-secreting capacity due to high expression of NKG2A.

Changes of ADAM metallopeptidase domain 23 expression in hepatic fibrosis of mice infected with Echinococcus multilocularis

ZHANG Hongbin, SHI Bowen, ZHENG Xue, LI Xiaotong, XUE Junlong, YANG Ning, BI Xiaojuan, LIN Renyong

2025, 43(6):  777-785.  doi:10.12140/j.issn.1000-7423.2025.06.005

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Objective To investigate the changes of ADAM metallopeptidase domain 23 (ADAM23) expression during the progression of hepatic fibrosis in mice infected with Echinococcus multilocularis (Em). Methdos Thirty-six C57BL/6 mice were randomly divided into six groups, of 6 mice each group, including the 1-month sham group, 1-month Em infection group, 3-month sham group, 3-month Em infection group, 6-month sham group, and 6-month Em infection group. Mice in the Em infection groups were injected with Em protoscolex suspensions (2 000 protoscoleces/mouse) via the portal vein, while animals in the sham groups received an equal volume of physiological saline. Mouse liver tissues were sampled 1, 3 and 6 months post-infection. Hepatic lobule structure, inflammatory cell infiltration, granuloma formation, and collagen deposition were observed by HE and Sirius red staining, and target protein expression was quantified using immunohistochemical staining. Mouse liver tissues from the 3-month sham and 3-month Em infection groups were sampled for proteomic analyses, and the translational and transcriptional expression of ADAM23, α-smooth muscle actin (α-SMA), and collagen type 1 alpha 1 chain (COL1A1) was determined in mice using Western blotting and RT-qPCR assays. Human hepatic stellate cells (HSCs) were seeded onto 6-well plates at a density of 2 × 10⁵ cells/well and divided into the si-ADAM23 group, TGF-β1 + small interfering RNA (siRNA) group, TGF-β1 + si-ADAM23 group, siRNA group, and control group. Cells were transfected with 600 ng siRNA-ADAM23 in the si-ADAM23 group, received 10 ng TGF-β1 and 600 ng siRNA cells in the TGF-β1 + siRNA group, received 10 ng TGF-β1 and 600 ng siRNA-ADAM23 in the TGF-β1 + si-ADAM23 group, received 600 ng siRNA in the siRNA group, and received an equal volume of transfection reagents in the control group. The translational and transcriptional expression of ADAM23, α-SMA, and COL1A1 was detected in HSCs using Western blotting and RT-qPCR assays. Results HE staining showed hepatic lobule structural derangements with infiltration of a large number of inflammatory cells in Em-infected mice. Sirius red staining revealed that the areas of hepatic fibrosis were (2 167.00 ± 356.70) μm², (163.10 ± 25.41) μm², (3 792.00 ± 596.00) μm², (159.40 ± 3.97) μm², (5 436.00 ± 427.50) μm², (274.40 ± 38.01) μm² in the 1, 3, and 6-month Em infection groups and corresponding sham groups, respectively, with significant differences seen between the infection and sham groups (t = 14.71, 26.68 and 37.91; all P < 0.05). Proteomic analysis showed that the ADAM23 protein abundance was 9 250 ± 5 628 of in the 3-month Em infection group. Western blotting assay showed significant differences in ADAM23, α-SMA, and COL1A1 protein expression between the 1-, 3-, 6-month Em infection groups and their corresponding sham groups (t = 13.5, 15.8, 26.1; 4.37, 6.75, 15.82; 5.00, 8.65 and 21.45; all P < 0.05). RT-qPCR assay quantified significant differences in the relative mRNA expression of adam23, acta2, and col1a1 between the 1-, 3-, 6-month Em infection groups and their corresponding sham groups (t = 5.44, 6.44, 9.95; 4.13, 7.43, 25.10; 4.85, 22.60 and 39.50; all P values < 0.05). Immunohistochemistry showed significant differences in the area of positive ADAM23, α-SMA, and COL1A1 expression the 1-, 3-, 6-month Em infection groups and their corresponding sham groups (t = 26.68, 17.55, 52.07; 24.98, 36.70, 49.47; 16.24, 45.48 and 77.45; all P < 0.05). Spearman correlation analysis revealed that the area of positive ADAM23 expression positively correlated with the areas of positive α-SMA (r = 0.83, P < 0.05) and COL1A1 expression (r = 0.87, P < 0.05), and Sirius red staining (r = 0.88, P < 0.05) in liver samples of Em-infected mice. RT-qPCR assay quantified that the relative adam23 mRNA expression was 1.03 ± 0.14, 0.92 ± 0.09, and 0.23 ± 0.04 in HSCs in the control, siRNA, and si-ADAM23 groups, respectively, and higher relative adam23 mRNA expression was found in the siRNA group than in the si-ADAM23 group (t = 12.23, P < 0.05). Western blotting assay showed that the relative ADAM23 protein expression was 0.15 ± 0.01, 1.20 ± 0.11, and 0.41 ± 0.15 in the siRNA group, TGF-β1 + siRNA group, and TGF-β1 + si-ADAM23 group, statistically significant differences were observed both when comparing the siRNA group with the TGF-β1 + siRNA group, and when comparing the TGF-β1 + siRNA group with the TGF-β1 + si-ADAM23 group (t = 16.52 and 12.44, both P < 0.05), and the relative α-SMA protein expression was 0.31 ± 0.09, 1.21 ± 0.04, and 0.87 ± 0.01 in the siRNA group, TGF-β1 + siRNA group, and TGF-β1 + si-ADAM23, statistically significant differences were observed both when comparing the siRNA group with the TGF-β1 + siRNA group, and when comparing the TGF-β1 + siRNA group with the TGF-β1 + si-ADAM23 group (t = 27.85 and 10.67, both P values < 0.05), while the relative COL1A1 protein expression was 0.61 ± 0.16, 0.95 ± 0.02, and 0.49 ± 0.07 in the siRNA group, TGF-β1 + siRNA group, and TGF-β1 + si-ADAM23 group, statistically significant differences were observed both when comparing the siRNA group with the TGF-β1 + siRNA group, and when comparing the TGF-β1 + siRNA group with the TGF-β1 + si-ADAM23 group (t = 5.77 and 7.96, both P values < 0.05). Conclusion ADAM23 expression increases with the progression of Em infections and positively correlates with the degree of hepatic fibrosis. ADAM23 is a novel potential therapeutic target for hepatic fibrosis caused by Em infections.

Therapeutic efficacy of albendazole and pyronaridine for the treatment of cystic echinococcosis in sheep

YAN Mingzhi, ZHANG Yao, QI Wenjing, WANG Xue, ZHANG Guowu, WU Chuanchuan, TIAN Mengxiao, GENG An, WANG Weisi, DUAN Liping, SONG Tao, YOU Xihuo, LI Jun, ZHANG Wenbao

2025, 43(6):  786-792.  doi:10.12140/j.issn.1000-7423.2025.06.006

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Objective To evaluate the efficacy of albendazole (ABZ) and pyronaridine (PND) for the treatment of cystic echinococcosis (CE) in sheep. Methods B-mode ultrasonography was performed in sheep from the central Tianshan Mountains regions in Xinjiang Uygur Autonomous Region, and 18 sheep with typical CE characteristics and comparable prevalence were selected, and randomized into the control group, the ABZ group and PND group using a completely randomized design. Sheep in the ABZ group were fed baits containing ABZ (0.4 g ABZ in 1 g feed bait), and sheep in the PND group were fed baits containing PND (0.2 g PND in 1 g feed bait), while sheep in the control group were fed drug-free feed baits. Drugs were administered continuously for 30 days, followed by a 30-day observation period after drug discontinuation. Whole-liver scans were performed on sheep with CE before dosing and 30 days after drug discontinuation, and cyst size was recorded. Sheep hearts, livers, spleen, lungs, and kidneys were collected and weighed 30 days after drug discontinuation. Whole blood samples were collected for routine blood tests and blood biochemistry analysis. Liver cysts were sampled, and classified into fertile cysts, infertile cysts, and calcified cysts based on the cyst appearance and its contents. The number and weight of different types of cysts were recorded. Hepatic tissues were prepared into paraffin-embedded sections, subjected to hematoxylin-eosin (HE) staining for pathological examinations, and evaluated for scoring of inflammatory cell infiltration. All statistical analyses were performed using the software IBM SPSS Statistics 20. Multiple comparisons were conducted using one-way analysis of variance or nonparametric tests, while intergroup comparisons of cyst types were assessed using chi-square test. Results B-mode ultrasonography measured no significant difference in the diameter of liver cysts among the control group [(2.86 ± 0.94) cm], ABZ group [(2.58 ± 0.83) cm], and PND group [(2.65 ± 0.81) cm] prior to oral administration (F = 0.090, P > 0.05). The diameters of cysts were (0.41 ± 0.30) cm in the ABZ group and (0.85 ± 0.24) cm in the PND group 30 days after drug discontinuation, which were significantly smaller than in the control group [(3.03 ± 0.42) cm] (F = 110.398, P < 0.01). The sheep liver weights were (0.98 ± 0.20), (0.74 ± 0.16), and (0.61 ± 0.30) kg in the control group, ABZ group, and PND group 30 days after drug discontinuation, respectively (F = 3.989, P < 0.05). There were no significant differences in routine blood and biochemical parameters among the control, ABZ, and PND groups (all P > 0.05). The gross cyst weights were (0.02 ± 0.05) kg and (0.05 ± 0.08) kg in the ABZ and PND groups 30 days after drug discontinuation, which were significantly lower than in the control group [(0.17 ± 0.06) kg] (F = 7.835, P < 0.01). The proportions of fertile cysts, infertile cysts, and calcified cysts were 63.16% (48/76), 17.10% (13/76), and 19.74% (15/76) in the control group, and 23.08% (12/52), 1.92% (1/52), and 75.00% (39/52) in the ABZ group, respectively (χ² = 39.439, P < 0.01), with reduced proportions of fertile and infertile cysts and an increased proportion of calcified cysts in the ABZ group. The proportions of fertile cysts, infertile cysts, and calcified cysts were 31.03% (18/58), 6.90% (4/58), and 62.07% (36/58) in the PND group, which showed significant differences relative to the control group (χ² = 25.083, P < 0.01), with an reduced proportion of fertile cysts and an increased proportion of calcified cysts in the PND group. The scores of inflammatory cell infiltration were 1.06 ± 0.25 and 1.50 ± 0.28 in the ABZ and PND groups 30 days after drug discontinuation, which were both significantly lower than in the control group (2.72 ± 0.44) (F = 39.780, P < 0.01). Conclusion Oral administration of ABZ or PND may reduce the number of fertile cysts in livers of sheep with CE and alleviate the severity of inflammatory cell infiltration, indicating that both chemicals are effective against sheep CE.

Epidemiological characteristics on liver fibrosis among individuals with a history of schistosome infection in lake-marsh type endemic area

PAN Youwen, LI Shizhen, HUANG Song, LIU Xiong, LI Zhu, YANG Meikai, LI Guocui, ZHU Hong, YUAN Yi, ZHANG Lijuan, ZHANG Xia

2025, 43(6):  793-799.  doi:10.12140/j.issn.1000-7423.2025.06.007

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Objective To investigate the demographic characteristics and prevalence of liver fibrosis among individuals with a history of schistosomiasis in lake-marsh type endemic area, providing evidence for the early prevention, detection, and intervention of advanced schistosomiasis. Methdos Individuals registered as previously infected with Schistosoma japonicum in Jiangling County, Hubei Province between 2008 and 2012 were selected through cluster sampling. A structured questionnaire was used to collect demographic data. Liver fibrosis was assessed by Doppler ultrasound. The four liver fibrosis indicators were detected using chemiluminescence assay, and the liver function biochemical indicators were measured with the automated biochemical analyzer. The anti-schistosome antibodies were detected using the indirect hemagglutination assay (IHA). Statistical analyses were performed using R software version 4.0.0. Chi-square tests were applied to categorical variables, and one-way ANOVA was used for continuous variables. Results A total of 514 individuals with a history of schistosome infection were included, the ratio of male/female was 1.28∶1, with a mean age of (61.63 ± 9.63) years. Ultrasound results indicated that 28.21% (145/514) individuals had normal liver parenchyma, 36.19% (186/514) had mild liver fibrosis, and 35.60% (183/514) had moderate to severe liver fibrosis. Statistically differences in liver fibrosis severity were observed across different sex and age groups (χ² = 14.48, 39.34; both P < 0.01). The proportions of abnormal liver morphology were 13.97% (20/145), 28.49% (53/186), and 34.97% (64/183) in individuals with normal liver parenchyma, mild liver fibrosis, and moderate to severe liver fibrosis, respectively (χ² = 19.07, P < 0.01). Mean spleen thickness of individuals with normal liver parenchyma, mild liver fibrosis, and moderate to severe liver fibrosis were (29.72 ± 4.62), (30.37 ± 5.00), and (31.34 ± 4.88) mm, respectively (F = 4.58, P < 0.05); and the portal vein diameters were (10.47 ± 0.80), (10.32 ± 0.76), and (10.58 ± 0.87) mm, respectively (F = 9.59, P < 0.01). The abnormal rates of hyaluronidase (HA) among the individuals with normal liver parenchyma, mild liver fibrosis, and moderate to severe liver fibrosis were 18.62% (27/145), 28.50% (53/186), and 39.34% (72/183), with a statistically difference (χ² = 16.84, P < 0.01). There were no statistically differences observed in other liver fibrosis indicators (procollagen Ⅲ, laminin, or collagen type Ⅳ) or liver function biochemical indicators (total bilirubin, total protein, albumin, alanine aminotransferase, or aspartate aminotransferase). IHA results indicated that 94 individuals tested positive for anti-schistosome antibodies, corresponding to an overall antibody positivity rate of 18.29%; the antibody positivity rate among the individuals with normal liver parenchyma, mild liver fibrosis, and moderate to severe liver fibrosis were 15.17% (22/145), 20.97% (39/186), and 18.03% (33/183), respectively (χ² = 1.84, P > 0.05). No cases were found to be positive on parasitological examination. Conclusion Individuals with a history of schistosomiasis in marshland-endemic regions exhibit a high prevalence of liver fibrosis and antibody positivity. The severity of fibrosis correlates with abnormal liver morphology, increased spleen thickness, portal vein dilation, and elevated HA levels. Enhanced monitoring, targeted management, and early intervention in this population are essential to reduce the burden of advanced schistosomiasis.

Evaluation of the effectiveness of the integrated clonorchiasis control programme in Guangdong Province

ZHANG Guanting, ZHANG Xianchang, ZHANG Qiming, LUO Le, ZENG Qingsheng, FANG Yueyi, LIAO Yuhuang, MAO Qiang, LIU Jiahui, DENG Zhuohui, WU De, CHEN Jingdiao

2025, 43(6):  800-805.  doi:10.12140/j.issn.1000-7423.2025.06.008

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Objective To investigate the effectiveness of the integrated clonorchiasis control programme in hyperendemic areas of Guangdong Province (Zhongshan City and Xinhui District of Jiangmen City), so as to provide reproducible and scalable integrated control experiences in other disease-affected regions. Methdos Integrated clonorchiasis control measures were implemented in Zhongshan City and Xinhui District of Jiangmen City during the period from 2019 to 2023, including online and offline health education, environmental improvements focusing on construction of sanitary toilets and provision of safe water in the text of new rural construction, albendazole given to Clonorchis sinensis-infected individuals, improvements of professional capabilities among disease control teams by means of training and hands-on practices. An administrative village (community) was selected from each township (street) in eastern, western, southern, northern and central parts according to geographical locations in two implementation areas after implementation of the integrated control measures. A total of 200 residents at ages of over 3 years were sampled using the cluster sampling method for detection of the prevalence of human clonorchiasis. And use the findings from the most recent population infection survey conducted prior to the project’s initiation as baseline data for comparison. In addition, a total of 150 individuals were randomly sampled from residents subjected to surveys of the prevalence of C. sinensis human infections in 2019 prior to the programme and 2023 at the final evaluation for questionnaire surveys of healthy human behaviors. Results There were 227 administrative villages received health education pertaining to clonorchiasis control knowledge in Zhongshan City, and 224 in Xinhui District, with a coverage rate of 100% in both areas from 2019 to 2023. The awareness of human clonorchiasis control knowledge was 96.0% (261/272) in Zhongshan City and 98.0% (245/250) in Xinhui District in 2023, which increased by 60.0% and 40.0% relative to 2019 before implementation of the programme [60.0% (150/250) and 70.0% (105/150)], respectively. In terms of environmental improvements, 89 850 new sanitary toilets were built and 89 850 households with access to safe water were provided in Zhongshan City, and 17 094 new sanitary toilets were built and 16 968 households with access to safe water were provided in Xinhui District, with 100% coverage rates of sanitary toilets and safe water supply in both areas. There were 10 424 and 22 427 C. sinensis-infected individuals identified through community-based surveys on human infections and standardized clonorchiasis clinics in medical institutions in Zhongshan City and Xinhui District, and 10 185 and 16 821 individuals given deworming chemicals, with deworming coverage rates of 97.71% and 75.00%, respectively. The proportion of professionals receiving training increased from 91.67% (66/72) to 100% (103/103) in Zhongshan City and from 52.94% (90/170) to 98.05% (201/205) in Xinhui District. The prevalence of C. sinensis infections reduced from 37.28% (400/1 073) at baseline to 11.52% (131/1 137) at the final evaluation in Zhongshan City (χ² = 625.24, P < 0.05) and from 34.18% (499/1 460) to 10.20% (102/1 000) in Xinhui District following implementation of 5-year integrated control programmes (χ² = 188.35, P < 0.05), respectively. Conclusion Through systematic interventions, Zhongshan City and Xinhui District in Guangdong Province significantly reduced the infection rate of C. sinensis, enhanced public awareness of prevention knowledge, and improved environmental sanitation, providing replicable and scalable experiences for comprehensive control in other regions.

Clinical characteristics of fascioliasis and primary liver cancer patients

HUANG Lihua, LI Nan, GU Wei

2025, 43(6):  806-813.  doi:10.12140/j.issn.1000-7423.2025.06.009

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Objective To analyze the clinical characteristics of fascioliasis and primary liver cancer patients, so as to provide insights into early differential diagnosis of these two diseases. Methdos The clinical data of 110 fascioliasis patients and 110 primary liver cancer patients admitted to The First Affiliated Hospital of Dali University during the period from January 1, 2014, to December 31, 2024 were collected, and the clinical characteristics, laboratory parameters, imaging features, and pathological manifestations were compared between the two groups. Comparisons of measurement data were done using t test or Mann-Whitney U test, while comparisons of count data were conducted with chi-square test or Fisher’s exact probability test. Results The onset date of fascioliasis patients was primarily distributed during the period from July to November, and the onset date of primary liver cancer patients was evenly distributed from January to December, without a clear-cut temporal distribution pattern. Fascioliasis patients were mainly from Dali Bai Autonomous Prefecture (62 cases, 56.4%), Lijiang City (15 cases, 13.6%), and Pu’er City (13 cases, 11.8%), and primary liver cancer patients mainly came from Dali Bai Autonomous Prefecture (84 cases, 76.4%), Nujiang Lisu Autonomous Prefecture (10 cases, 9.1%), and Baoshan City (9 cases, 8.2%). Fascioliasis patients had a disease course (from disease onset to healthcare seeking) of approximately 3 days to one year, which was longer than primary liver cancer patients (2 to 60 days). There were significant differences between fascioliasis and primary liver cancer patients in terms of age, gender distribution, proportion of a medical history of chronic viral hepatitis B, proportion of smoking, proportion of alcohol consumption, proportion of fever, proportion abdominal pain, proportion of abdominal distension, proportion of weight loss, alpha-fetoprotein level, carcinoembryonic antigen level, carbohydrate antigen 125 level, carbohydrate antigen 153 level, carbohydrate antigen 199 level, fibrinogen level, white blood cell count, neutrophils percentage, eosinophils percentage, absolute eosinophil count, hemoglobin level, platelet count, total bilirubin, direct bilirubin, indirect bilirubin, alanine aminotransferase activity, aspartate aminotransferase, gamma-glutamyl transpeptidase activity, and albumin level (t = 10.73, χ² = 48.6, 186.72, 41.56, 19.70, 42.49, 16.37, 5.74, 14.08, Z = -10.65, -3.03, -7.99, -6.05, -6.30, -2.65, -5.63, -10.20, -12.00, -12.05, -2.76, -7.75, -7.70, -8.00, -6.69, -4.79, -9.02, -4.91, -2.81, all P < 0.05). CT scans displayed multiple round-like low-density shadows in fascioliasis patients’ livers, which appeared a clustered distribution pattern; however, no enhancement was seen in the arterial phase following contrast enhancement. Contrast, abdominal CT scans displayed nodular low-density lesions with obvious enhancement in the arterial phase following contrast enhancement among primary liver cancer patients. In addition, the major pathological manifestations of fascioliasis patients included irregular tunnel-like necrosis, eosinophilic abscesses, and Charcot-Leyden crystals in the liver, while all primary liver cancer patients had a pathological type of hepatocellular carcinoma. Conclusion There are significant differences between fascioliasis and primary liver cancer patients in terms of time of disease onset, age distribution, gender ratio, laboratory parameters, imaging findings, and pathological characteristics. Combining epidemiological and clinical features may provide insights into early differential diagnosis.

Role of m⁶A modification in mediation of TRPM8 in macrophages following Toxoplasma gondii infection and its underlying mechanisms

LIN Shuqing, SHAO Tianye, ZHANG Zixin, LIU Xinjian, ZHANG Rong, WANG Yong, QIU Jingfan

2025, 43(6):  814-820.  doi:10.12140/j.issn.1000-7423.2025.06.010

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Objective To investigate the role of N⁶-methyladenosine (m⁶A) modification in regulation of transient receptor potential cation channel subfamily M member 8 (TRPM8) protein in macrophages following Toxoplasma gondii infection and unravel its underlying mechanisms. Methdos Transcriptome sequencing (RNA-seq) and methylated RNA immunoprecipitation sequencing (MeRIP-seq) data were sourced from the NCBI Gene Expression Omnibus (GEO) dataset GSE288205. Transient receptor potential (TRP) channel protein family-coding genes were subjected to bioinformatics analyses with the OmicStudio platform, and volcano plots for m⁶A modifications of TRP channel-associated factor 1 (TCAF1) and TRPM8 mRNA were generated based on MeRIP-seq data using ggplot2 in the RStudio package. THP-1-derived macrophages, human peripheral blood mononuclear cells (PBMCs) and mouse RAW264.7 cells were seeded onto 6-well plates at a density of 1 × 10⁶ cells per well. Cells infected with T. gondii RH strain tachyzoites at a cell-to-parasite ratio of 1∶1 served as the infection group, while cells in the control group received an equal volume of parasite-free medium. Total RNA was extracted from cells using the TRIzol reagent, reversely transcribed into cDNA. The relative mRNA expression of TRPM8 (human) and Trpm8 (mouse) was quantified in cells using quantitative fluorescent real-time PCR (qPCR) assay, and the relative TCAF1 and TCAF2 mRNA levels were detected in THP-1-derived macrophages with and without infected with T. gondii tachyzoites infections using qPCR assay. Intracellular Ca²⁺ influx was detected in cells using flow cytometry. THP-1-derived macrophages, fat mass and obesity-associated protein (FTO)-knockdown THP-1-derived macrophages (shFTO-THP-1), and THP-1-derived macrophages carrying non-targeted control shRNA (shNC-THP-1) were seeded onto 6-well plates at a density of 1 × 10⁶ cells per well. Cells in the infection group were infected with T. gondii RH strain tachyzoites at a cell-to-parasite ratio of 1∶1, while cells in the control group received an equal volume of parasite-free medium. Changes in m⁶A modification on TCAF1 mRNA in THP-1-derived macrophages were validated using m⁶A methylated RNA immunoprecipitation-real time quantitative PCR (m⁶A-IP-qPCR) assay. The expression of TCAF1 mRNA binding the m⁶A recognition protein YTH domain family protein 2 (YTHDF2) was detected in THP-1-derived macrophages infected with T. gondii tachyzoites using RNA immunoprecipitation (RIP) assay. The rate of TCAF1 mRNA degradation was compared between the shFTO-THP-1 and shNC-THP-1 groups using qPCR assay 0, 2, 4, and 6 hours post-treatment with actinomycin D. Differences of means between groups were tested for statistical significance with independent-sample Student’s t-test. Results RNA-seq revealed significant downregulation of TRP channel protein-coding genes except transient receptor potential cation channel subfamily C member 4-related protein, which was upregulated. Notably, TRPM8 exhibited the most pronounced downregulation, with its transcription level at 0.16-fold of the control group. qPCR assay quantified lower TRPM8 (human) and Trpm8 (mouse) mRNA expression in THP-1-derived macrophages [(0.445 ± 0.118) vs. (1.000 ± 0.036); t = 7.828, P < 0.05], PBMCs [(0.302 ± 0.040) vs. (1.042 ± 0.381); t = 3.345, P < 0.05], and RAW264.7 cells [(0.365 ± 0.234) vs. (1.004 ± 0.105); t = 4.312, P < 0.05] infected with T. gondii tachyzoites than in controls, and flow cytometry detected lower intracellular fluorescence intensities of Ca²⁺ in THP-1-derived macrophages [(19 500.0 ± 2 427.0) vs. (23 500.0 ± 2 64.6); t = 2.838, P < 0.05], PBMCs [(3 569.0 ± 313.9) vs. (13 050.0 ± 1 072.0); t = 16.970, P < 0.05], and RAW264.7 cells [(6 513.0 ± 348.0) vs. (7 683.0 ± 245.2); t = 4.762, P < 0.05] infected with T. gondii tachyzoites than in controls. qPCR assay quantified lower TCAF1 mRNA expression in THP-1-derived macrophages [(0.617 ± 0.132) vs. (1.005 ± 0.115); t = 3.832, P < 0.05] infected with T. gondii tachyzoites than in controls, and no significant differences in TCAF2 mRNA expression between the infection and control groups [THP-1-derived macrophages: (0.973 ± 0.030) vs. (1.015 ± 0.209), t = 0.343, P > 0.05]. Volcano plots of MeRIP-seq data displayed that m⁶A modification in the 3′ untranslated region (3′UTR) of TCAF1 mRNA was upregulated by 552 folds in THP-1-derived macrophages infected with T. gondii tachyzoites than in controls, and m⁶A-IP-qPCR assay detected a higher m⁶A modification level in the 3′ UTR region of TCAF1 mRNA in THP-1-derived macrophages infected with T. gondii tachyzoites than in controls [(4.794 ± 0.854) vs. (1.000 ± 0); t = 7.696, P < 0.05]. YTHDF2-RIP analysis showed a higher transcriptional level of TCAF1 mRNA binding to YTHDF2 in THP-1-derived macrophages infected with T. gondii tachyzoites than in controls [(2.423 ± 0.782) vs. (1.010 ± 0.180); t = 3.048, P < 0.05]. In addition, a higher rate of TCAF1 mRNA degradation was seen in shFTO-THP-1 cells infected with T. gondii tachyzoites than in shNC-THP-1 cells infected with T. gondii tachyzoites. Conclusion m⁶A modification contributes to downregulation of the cold receptor TRPM8 in both human and mouse macrophages following T. gondii infection.

Development of a recombinase polymerase amplification-CRISPR/Cas12a assay for detection of Leishmania major

CHEN Xinyi, ZHANG Bingke, LANG Jiashu, XU Huiyan, LI Junqiang, WU Longfei, ZHANG Sumei, ZHANG Longxian

2025, 43(6):  821-826.  doi:10.12140/j.issn.1000-7423.2025.06.011

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Objective To develop a rapid and accurate assay for detection of Leishmania major based on the recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 12a (Cas12a) system, so as to improve the diagnostic efficiency of cutaneous leishmaniasis. Methdos Based on the internal transcribed spacer 1 (ITS1) gene sequence of L. major (GenBank accession number: KM052753.1) retrieved from NCBI as the target sequence, RPA primers, CRISPR RNA (crRNA), and single-stranded DNA (ssDNA) reporter probe were designed. An RPA-CRISPR/Cas12a assay was developed by screening RPA primers and optimizing the RPA reaction temperature. Genomic DNA was extracted from L. major, and the ITS1 gene sequence was amplified. The amplification product was cloned into the pUC18 plasmid to construct a recombinant plasmid, which was serially diluted to 8 concentrations of 2.3 × 10⁶, 2.3 × 10⁵, 2.3 × 10⁴, 2.3 × 10³, 2.3 × 10², 2.3 × 10¹, 2.3 × 10⁰, and 2.3 × 10⁻¹ copies/μl as templates for RPA-CRISPR/Cas12a assay and PCR assay to evaluate the detection sensitivity of the plasmid. L. major promastigotes were added to 200 μl of canine whole blood samples at a concentration of 1.8 × 10⁷ parasites/ml, and genomic DNA was extracted, and serially diluted into 4 concentrations of 1.8 × 10², 1.8 × 10¹, 1.8 × 10⁰, and 1.8 × 10⁻¹ parasites/ml as templates for RPA-CRISPR/Cas12a assay to evaluate the detection sensitivity of parasites. Genomic DNA from L. major, L. infantum, Toxoplasma gondii, Babesia gibsoni, Neospora caninum, and Anaplasma phagocytophilum were used as templates for the RPA-CRISPR/Cas12a assay to evaluate the detection specificity. Repeatability of the RPA-CRISPR/Cas12a assay was assessed by performing 5 independent experiments with 3 replicates each using the 2.3 × 10⁴ copies/μl recombinant plasmid as the template. To compare the detection performance of the RPA-CRISPR/Cas12a and the PCR assay, L. major promastigotes were added to 10 samples of 200 μl canine whole blood at a concentration of 1.8 × 10⁵ parasites/ml for RPA-CRISPR/Cas12a and PCR assays, with negative controls set simultaneously. Results The RPA-CRISPR/Cas12a method was established using the F7/R7 primer pair, with optimized reaction conditions set as 41 ℃ for 25 minutes. The limits of detection (LOD) of RPA-CRISPR/Cas12a and PCR assays were 2.3 copies/μl and 2.3 × 10³ copies/μl of the recombinant plasmid, and the LOD of the RPA-CRISPR/Cas12a assay was 1.8 promastigotes/ml of parasites; however, this assay showed no cross-reactivity of L. major with L. infantum, T. gondii, B. gibsoni, N. caninum, or A. phagocytophilum. Repeatability testing showed stable and consistent fluorescence intensities across different batches, with a relative standard deviation of 4.27% (< 5%). Both RPA-CRISPR/Cas12a and PCR assays accurately detected all 10 spiked canine whole blood samples containing L. major promastigotes, achieving the same detection rate and a 100% coincidence rate. Conclusion The RPA-CRISPR/Cas12a assay developed in this study is rapid, sensitive, specific, and reproducible for detection of L. major, which may provide reliable technical supports for diagnosis of L. major infections.

Mechanism underlying human airway epithelial cell inflammation triggered by dust mite allergen Der f 1

JIANG Shangde, ZANG Xiao, MEI Wei, MA Lianzheng, RAO Lina, HONG Shanchao, WANG Wei

2025, 43(6):  827-834.  doi:10.12140/j.issn.1000-7423.2025.06.012

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Objective To decipher the mechanisms underlying human airway epithelial cell inflammation induced by the major allergen Der f 1 of dust mite. Methdos BEAS-2B cells were seeded onto 96-well plates at a density of 5 × 10³ cells per well and divided into three groups, including the Der f 1 group, Der f 1 + ferrostatin-1 (Fer-1) group, and negative control group. Cells in the Der f 1 group were treated with 40 μl of Der f 1 (100 μg/ml), and cells in the Der f 1 + Fer-1 group was pretreated with 4 μl Fer-1 (1 000 μmol/ml) 2 hours prior to Der f 1 treatment, while cells in the negative control group were given an equivalent volume of nuclease-free water. After 24 hours of incubation, cell viability was assessed in the Der f 1 group, Der f 1 + Fer-1 group and negative control group using the CCK-8 assay, and concentrations of interleukin-6 (IL-6), thymic stromal lymphopoietin (TSLP), and IL-33 were measured in the cell culture supernatants with enzyme-linked immunosorbent assay (ELISA). For Western blotting assay, BEAS-2B cells were seeded onto 6-well plates at a density of 5 × 10⁵ cells per well, and assigned into three groups, including the Der f 1 group, Der f 1 + Fer-1 group, and negative control group. Total protein was extracted from BEAS-2B cells, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and probed with primary antibodies against glutathione peroxidase 4 (GPX4, 1︰2 000 dilution) and acyl-CoA synthetase long-chain family member 4 (ACSL4, 1︰2 000 dilution), followed by incubation into horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG secondary antibody (1︰10 000 dilution). The GPX4 and ACSL4 protein expression was determined in cells using Western blotting assay. To assess ferroptosis-related indicators, BEAS-2B cells were seeded onto 24-well plates at a density of 5 × 10⁴ cells per well and assigned into three groups, including the Der f 1 group, Der f 1 + Fer-1 group, and negative control group. Intracellular fluorescence intensity of Fe²⁺ and reactive oxygen species (ROS) were measured under a fluorescence microscope, and the ratio of red to green fluorescence intensity was estimated following lipid peroxidation (LPO) staining. For colorimetric assay, BEAS-2B cells were seeded onto 6-well plates at a density of 5 × 10⁵ cells per well and assigned into three groups, including the Der f 1 group, Der f 1 + Fer-1 group, and negative control group, and malondialdehyde (MDA) content was measured using a colorimetric assay. In addition, BEAS-2B cells were seeded onto 6-well plates at a density of 5 × 10⁶ cells per well and assigned into three groups, including the Der f 1 group, Der f 1 + Fer-1 group, and negative control group, and mitochondrial morphology was examined under a transmission electron microscope. All statistical analyses were performed using the software GraphPad Prism 8.0.1, and differences of means among three groups were tested for statistical significance with one-way analysis of variance (ANOVA). Results The cell viability was significantly lower in the Der f 1 group (0.79 ± 0.03) than in the negative control (1.07 ± 0.08) (t = 6.663, P < 0.01), and co-treatment with Fer-1 restored cell viability to 0.94 ± 0.03 (t = 6.694, P < 0.01). ELISA showed that the IL-6 concentration in the supernatant of the Der f 1 group was (117.30 ± 21.32) pg/ml, which was higher than that of the negative control group (50.07 ± 5.82) pg/ml (t = 5.279, P < 0.01). The IL-6 concentration in the supernatant of the Der f 1 + Fer-1 group was (50.31 ± 12.28) pg/ml, which was lower than that of the Der f 1 group (t = 4.721, P < 0.01). The TSLP concentration in the supernatant of the Der f 1 group was (10.00 ± 2.37) pg/ml, which was higher than that of the negative control group (3.81 ± 0.92) pg/ml (t = 4.223, P < 0.05). The TSLP concentration in the supernatant of the Der f 1 + Fer-1 group was (4.41 ± 1.59) pg/ml, which was lower than that of the Der f 1 group (t = 3.399, P < 0.05). The IL-33 concentration in the supernatant of the Der f 1 group was (24.18 ± 2.53) pg/ml, which was higher than that of the negative control group (12.09 ± 2.08) pg/ml (t = 6.39, P < 0.01). The IL-33 concentration in the supernatant of the Der f 1 + Fer-1 group was (15.76 ± 1.39) pg/ml, which was lower than that of the Der f 1 group (t = 5.045, P < 0.01). Western blotting determined lower relative GPX4 protein expression in the Der f 1 group (0.38 ± 0.08) than in the negative control group (1.00 ± 0.00) (t = 13.21, P < 0.01), and higher GPX4 protein expression in the Der f 1 + Fer-1 group (0.72 ± 0.08) than in the Der f 1 group (t = 5.122, P < 0.01). Conversely, ACSL4 protein expression was upregulated in the Der f 1 group (1.74 ± 0.12 vs. 1.00 ± 0.00; t = 10.65, P < 0.01) and reduced by Fer-1 treatment (1.33 ± 0.13, t = 4.094, P < 0.05). Fluorescence microscopy showed increased intracellular fluorescence intensity of Fe²⁺ in the Der f 1 group (37.19 ± 5.42 vs. 11.93 ± 0.54; t = 8.035, P < 0.01), which was significantly lowered by Fer-1 treatment (13.16 ± 1.89, t = 7.253, P < 0.01). The intracellular ROS fluorescence intensity in the Der f 1 group was 13.48 ± 3.36, which was higher than that in the negative control group (6.80 ± 0.60, t = 3.386, P < 0.05). The fluorescence intensity in the Der f 1 + Fer-1 group was 7.35 ± 0.42, which was lower than that in the Der f 1 group (t = 3.134, P < 0.05). The intracellular LPO fluorescence intensity in the Der f 1 group was 3.91 ± 1.65, which was higher than that in the negative control group (0.31 ± 0.13, t = 3.775, P < 0.05). The LPO fluorescence intensity in the Der f 1 + Fer-1 group was 0.80 ± 0.15, which was lower than that in the Der f 1 group (t = 3.262, P < 0.05). The colorimetric assay results showed that the intracellular MDA content in the Der f 1 group was (5.57 ± 1.66) nmol/mg, which was higher than that in the negative control group (2.18 ± 0.51) nmol/mg (t = 3.393, P < 0.05). The MDA content in the Der f 1 + Fer-1 group was (2.24 ± 0.38) nmol/mg, which was lower than that in the Der f 1 group (t = 3.4, P < 0.05). Transmission electron microscopy displayed well mitochondrial status, clear mitochondrial boundary and many mitochondrial cristae in the negative control group, and Der f 1 exposure led to mitochondrial shrinkage, membrane thickening, and reduction or loss of cristae. These structural alterations were markedly attenuated in the Der f 1 + Fer-1 group, including weakening of mitochondrial shrinkage, membrane thickening and reduced numbers of mitochondrial cristae. Conclusion Der f 1 induces ferroptosis in human airway epithelial cells through disrupting iron homeostasis, suppressing antioxidant defense, and promoting lipid peroxidation, thereby potentiating the release of IL-6, TSLP, and IL-33 to exacerbate airway inflammation.

Morphological and molecular identification of a new record species of Heterakis from Rattus norvegicus in China

JÜ Huidong, LI Yang, JIAN Rui, DU Xin, SUN Jing, YAN Zihan, MENG Jiaxuan, LIU Shanshan, ZHAI Shiyong

2025, 43(6):  835-842.  doi:10.12140/j.issn.1000-7423.2025.06.013

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Objective To characterize the nematode specimens collected from large intestines of wild Rattus norvegicus in Shijiazhuang City by means of morphological and molecular biological techniques assays. Methdos Nematodes were sampled from large intestines of wild R. norvegicus in Gaocheng District, Shijiazhuang City, Hubei Province, and the morphological characteristics of nematodes were examined using light and scanning electron microscop to identify the nematode species. A female worm and a male worm were sampled for extraction of genomic DNA, and internal transcribed spacer 1 (ITS-1) and mitochondrial cytochrome c oxidase subunit 1 (cox1) genes were amplified. Following sequencing of amplification products, phylogenetic trees were created using Bayesian inference and maximum likelihood methods. Results A total of 35 nematodes were identified, including 16 males and 19 females. The worms were small and cylindrical, tapered at both ends. The thin cuticles exhibited fine and transverse striations and lateral alae, with the latter originating from the pharyngeal region. The head featured three well-developed lips, including a dorsal lip with two papillae and two ventrolateral lips, each with one papilla. The anterior esophagus was narrow and cylindrical, expanding posteriorly into a bulb. The body length was 7.083 mm to 9.708 mm in males and 9.541 mm to 12.208 mm in females. Males were further characterized by two spicules of approximately equal length (0.250 mm to 0.310 mm) and nine pairs of caudal papillae. The vulva of female worms was situated in the posterior body region, surrounded by 3 to 5 cuticular folds. The tail terminated in a slender point adorned with a pair of terminal papillae and the distance from the anus to the tail tip measured 0.916 mm to 1.083 mm. The nematode eggs were elliptical in shape, with a length measuring 0.050 mm to 0.058 mm and a width measuring 0.033 mm to 0.041 mm. Sequencing revealed that the sequence of the ITS-1 gene was 430 bp in length (GenBank accession No. PX622390), and shared 99.91% similarity with Heterakis dahomensis (GenBank accession No. JX845277) and 82.4% to 90.3% similarity with other 6 Heterakis species. The sequence of the cox1 gene was 345 bp in length (GenBank accession No. PX622389), and shared 82.0% to 96.7% similarity with other 7 Heterakis species. Phylogenetic analysis based on the cox1 sequence of Heterakis revealed that nematode obtained from this study was closely related to H. spumosa but independent of each other, and was distantly related to other species of Heterakis. Phylogenetic analysis based on the ITS-1 gene sequence showed that the nematode sampled from this study was most closely related to H. dahomensis, and was clustered with Heterakis spumosa into a clade, but was distantly to other species of Heterakis. Conclusion Based on morphological characteristics, gene sequencing, and phylogenetic analysis, the nematode obtained from this study was identified as H. dahomensis, which is a newly recorded species in China.

Identification and sequence analysis of Patagifer bilobus derived from Platalea leucorodia

KUANG Shaoxiang, SHAO Ruiqing, FENG Yingying, MIAO Lujun, SUN Zhiyong

2025, 43(6):  842-848.  doi:10.12140/j.issn.1000-7423.2025.06.014

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Objective To identify the species of 10 trematodes isolated from a dead adult Platalea leucorodia that had been rescued in the Poyang Lake area, and to analyze the sequence characteristics and genetic and evolutionary relationships of mitochondrial DNA (mtDNA)-encoded cytochrome C oxidase subunit I (cox1) gene and ribosomal DNA (rDNA)-encoded internal transcribed spacer (ITS) sequence. Methdos A trematode worm body-shaped structure isolated from the duodenum of P. leucorodia was observed under an optical microscope, and three worms were selected for hydrochloric acid carmine staining and preliminary identification of genus and species under a microscope. In addition, genomic DNA was extracted from another three selected worm samples for PCR amplification of cox1 and ITS sequences, and the amplification products were sequenced. Sequence alignment and homology search were performed in the GenBank database, and repetitive DNA and base content were estimated. Schistosoma japonicum cox1 and ITS sequences were selected as outgroups to estimate intraspecies and interspecies genetic distances, and to build phylogenetic trees with the neighbor-joining (NJ) method. Results Isolated ten trematode worms were fleshy and thick in shape and pale red in color, with a mean length of (14.75 ± 1.89) mm. The cephalic collar was distinctly bilobed and wider than the worm body, and the head spines had blunt rod-shaped ends. Hydrochloric acid carmine staining revealed that the worm body had 58 to 59 head spines, and 4 angular spines. The cox1 gene and ITS sequences of the three trematodes have lengths of 426 and 985 bp, respectively. In the base composition of the cox1 gene sequence, the content of A + T (61.74%) was significantly higher than that of C + G (38.26%), indicating a marked AT bias. Conversely, in the base composition of the ITS gene sequence, the content of A + T (46.10%) was lower than that of C + G (53.90%), showing a GC preference. No repetitive sequences were detected in the cox1, ITS1, or 5.8S rRNA gene sequences of the trematodes, whereas a 10 bp inverted repetitive sequence was identified in the ITS2 gene sequence. The cox1 gene sequence of the trematode exhibited a 92.86% homology and the closest genetic distance (0.079) with Patagifer (GenBank accession number: OQ606511), and the ITS sequence of the trematode showed a 98.57% homology and the closest genetic distance (0.014) with P. bilobus (GenBank accession number: ON141921). The trematode shared the same small clade with trematodes of the genus Patagifer within the large clade of Echinostomatidae. Conclusion Morphological and molecular biological identification confirms the trematode isolated from P. leucorodia as P. bilobus.

REVIEWS

Impact of land use change on vector-borne disease transmission and its underlying mechanisms: a review

XIANG Jiangling, LI Qin, GUO Suying, XU Jing

2025, 43(6):  849-854.  doi:10.12140/j.issn.1000-7423.2025.06.015

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Land use change is fundamentally transforming earth’s ecosystems, resulting in biodiversity decline and ecosystem degradation, and becomes a critical driver of infectious diseases outbreaks. This article systematically reviews the advances in the impact of land use changes on vector-borne diseases transmission and its mechanisms, with an emphasis on the potential impacts of land use changes on vector redistribution and global disease transmission. Current evidence indicates that land use changes, such as deforestation, agricultural expansion, urbanization, and infrastructure development, remarkably reshape vector habitats and population dynamics, thereby increasing the risk of mosquito-borne, sandfly-borne, and snail-transmitted diseases outbreaks and expansion, and land use changes mainly affect disease transmission via habitat fragmentation, microclimate modification, host community reorganization, and human disturbances. To respond to these health challenges caused by land use changes, strategic priorities should focus on enhancing scientific researches, optimizing land use policies, improving surveillance and forecast systems, promoting ecological rehabilitations, and increasing public awareness.

Improved efficiency of the parasitic disease diagnostic laboratory network: practices and reflections from Sichuan Province

SHANG Jingye, ZHANG Zhongshuang, DENG Xiu, GONG Xi, LI Rongzhi, WU Zisong, QIN Zhiqiang, KANG Dianju, ZHONG Bo, LI Shizhu, LIU Yang

2025, 43(6):  855-861.  doi:10.12140/j.issn.1000-7423.2025.06.016

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As the management of major parasitic diseases gradually stepped into elimination and consolidation phases in China, the laboratory-based surveillance system encounters a strategic need of transforming from case identification to validation of transmission interruption and early warning of a risk of imported cases. Using the construction of the parasitic disease diagnostic laboratory network in Sichuan Province as a case, this paper summarizes the exploratory experience in achieving province-wide coverage of the province-level reference laboratories, building a tiered network architecture, and ensuring the quality of surveillance data, analyzes the main bottlenecks that constrain improvements in network effectiveness under the new circumstances and proposes a systemic development pathway through four dimensions, including network coordination, quality control assurance, technological innovation, and data empowerment. Practices from Sichuan Province demonstrate that promoting the transformation of the laboratory network from a single-disease control tool into a comprehensive, regional public health resource is a critical support to the scientific elimination and long-acting consolidation of parasitic diseases, which provides a reference development path in similar regions.

SHORT COMMUNICATIONS

Development of an intelligent canine management system and its application in echinococcosis control

YOU Xihuo, SONG Yongwu, DUOJIE Renqing, XIN Youhong, CAIRANG Zhouzai, QIE Deli, GUO Xia, FAN Xingye, YUE Hong, ZHANG Qinqin, XU Bin, XIE Yinlu, ZHANG Wenbao, ZENG Xuhao, CHEN Wangkai, HUANGFU Liusheng, LI Wei, HUANG Fuqiang

2025, 43(6):  862-865.  doi:10.12140/j.issn.1000-7423.2025.06.017

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To improve the echinococcosis control effectiveness, an intelligent canine management system integrating network technology, smart medication box, and a cloud management platform was designed, and applied in Shaliuhe Town and Haergai Town, Gangcha County, Qinghai Province in July 2022 and June 2023, respectively. The positive rate of Echinococcus infection decreased from 6.25% (2/32) to 0 (0/36) in dog stool samples in Shaliuhe Town following 2 months of deployment of the smart medication box, and remained at 0 (0/36) after 16 months (Fisher’s exact test, P > 0.05). The positive rate of Echinococcus infection decreased from reduced from 9.10% (5/55) to 2.00% (1/50) in dog stool samples in Haergai Town following 2 months of deployment of the smart medication box (Fisher’s exact test, P > 0.05), and then, 50 dog stool samples were collected each month for seven months, with no positive samples detected (Fisher’s exact test, P < 0.01). These findings demonstrate that the system effectively solves the problems of a difficulty in dog management and a low compliance to medication deployment, remarkably improves the timeliness and standardization of deworming and significantly reduces workload of disease control personnels via release of reminders of automate medication and stool monitoring functions. The study developed a dog intelligent management system and validated its application effectiveness in precise prevention and control of echinococcosis, providing innovative solutions for parasitic disease prevention and control.

Trend of malaria epidemic situation and prevention and control strategies before and after the elimination of malaria in Baoshan City, Yunnan Province

YANG Hexian, HUANG Dongsheng, ZHANG Jianling, YIN Gang, MA Lingjuan

2025, 43(6):  865-870.  doi:10.12140/j.issn.1000-7423.2025.06.018

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To consolidate the achievements of malaria elimination and understand the epidemiological characteristics of malaria in Baoshan City, malaria endemic data in Baoshan City before malaria elimination (2011-2017) and after malaria elimination (2018-2024) were collated and analyzed. Malaria endemic data from January 1, 2011 to December 31, 2024 in Baoshan City were collected through the China Disease Prevention and Control Information System. Databases were established using the software Microsoft Office Excel 2011, and descriptive statistical analyses were performed using the software SPSS 26.0. A total of 1 725 malaria cases were reported in Baoshan City from 2011 to 2024, with the highest number of cases reported in 2011 (406 cases) and the lowest in 2022 (7 cases), showing an overall declining trend in the number of reported cases year by year. Before malaria elimination, a total of 1 459 cases (85.58%) were reported, while 266 cases (15.42%) were reported after malaria elimination. The predominant infecting Plasmodium species was P. vivax (1 500 cases, 86.96%), with no change in the distribution of infecting Plasmodium species before and after malaria elimination. Reported cases were mainly distributed in Tengchong City (1 154 cases, 66.90%), with no change in regional distribution before and after malaria elimination. Cases were reported every month from 2011 to 2024, with the highest cumulative number of cases reported in May (330 cases, 19.13%), and the peak reporting period was from April to July (1 029 cases, 59.65%). The reported cases were predominantly male (1 486 cases, 86.14%), with farmers being the main occupational group (1 343 cases, 77.85%), and the age group mainly distributed between 20 and 55 years old (1 556 cases, 90.20%). The main population affected before and after malaria elimination consisted of young and middle-aged male farmers. The reported cases were mainly imported (1 707 cases, 98.96%), including 1 449 cases imported from abroad and 276 cases imported domestically. Before malaria elimination, 98.77% (1 441/1 459) of the reported cases were imported, and 79.80% (1 165/1 459) were imported from abroad. After malaria elimination, all 266 reported cases were imported from abroad. The main source of imported infections from abroad was Myanmar. A total of 1 648 malaria cases were diagnosed by medical institutions at all levels within the jurisdiction of Baoshan City from 2011 to 2024. A total of 1 384 cases were diagnosed before malaria elimination, mainly confirmed cases (1 343 cases, 97.04%), with county-level Centers for Disease Control and Prevention being the main diagnostic units (598 cases, 43.21%). A total of 264 malaria cases were diagnosed after malaria elimination, all of which were laboratory-confirmed cases, with municipal and county-level general hospitals being the main diagnostic units (180 cases, 68.18%). Before malaria elimination, the shortest time interval from onset to seeking medical care for reported cases was 0-day, and the longest was 214-day. After malaria elimination, the shortest time interval was 0-day, and the longest was 17-day. The main prevention and control measures in Baoshan City focused on reducing and eliminating local infections before malaria elimination, and the prevention and control emphasis shifted to the timely detection of imported cases and the prevention of local re-transmission after malaria elimination. The survey results showed that Baoshan City had reported no locally infected cases for 11 consecutive years since 2014, indicating remarkable achievements in malaria elimination. However, due to its proximity to Myanmar, the situation regarding the prevention and control of imported malaria in Baoshan City remains severe.

CASE REPORT

Renal echinococcosis misdiagnosed as a renal cyst: A case report

REYIHANGULI Wushouer, YIMIGULI Yimin, BUWEIREBIYEMU Tuersunaili, MAIMAITIMING Mahemuti, MA Haihong

2025, 43(6):  871-873.  doi:10.12140/j.issn.1000-7423.2025.06.019

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A 41-year-old male, farmer from Kashi, Xinjiang, was presented to Kashi Prefecture Second People’s Hospital on May 15, 2025, with complaints of “intermittent left-sided lumbago for 3 months”. The patient had a clear history of long-term sheep herding, slaughtering, and close contacts with sheep. B ultrasound of the urinary system displayed a left renal cyst complicated by infections. Abdominal CT scanning displayed a round, low-density lesion in the left kidney, measuring approximately 14 cm × 11 cm, and contrast-enhanced CT scans showed no enhancement in the center of the lesion, mild progressive enhancements in the cyst wall and cyst wall nodules, compressed renal pelvis and renal artery, clear surrounding fat space, displaced pancreatic tail by compression, and a lower enhancement intensity of the left kidney than that of the contralateral side. The patient has been diagnosed with a left renal cyst. Based on imaging findings and clinical symptoms, the patient underwent retroperitoneal laparoscopic unroofing and decompression of left renal cyst under general anesthesia on May 21, 2025. Intraoperative findings included a grayish-white, tough cyst wall, clear cyst fluids within the cyst, and daughter cysts and protoscoleces in the cystic cavity. The procedure was converted to excision of left renal Echinococcus cyst. Pathological examination of cyst wall tissues and cystic fluids confirmed the diagnosis of cystic echinococcosis in the left kidney. Oral administration of albendazole was given at a dose of 400 mg twice daily on day 2 post-operation. The patient recovered well and was discharged from hospital on day 8 post-operation. He was instructed to continue albendazole treatment for at least 6 months and to undergo regular follow-up with ultrasonography and liver function tests. No recurrence had been observed until now.

Diagnosis and treatment of a case with visceral leishmaniasis complicated by severe pulmonary infection

JIN Xuan, LIU Yuanli, DENG Chunqing

2025, 43(6):  874-877.  doi:10.12140/j.issn.1000-7423.2025.06.020

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The patient was a 38-year-old female farmer from Xiangning County, Linfen City, Shanxi Province. She was diagnosed with nephrotic syndrome in 2016 and systemic lupus erythematosus in 2021. In October 2024, the patient was admitted to the Department of Rheumatology and Immunology due to recurrent fever. Laboratory tests revealed pancytopenia, inverted albumin-globulin ratio, elevated IgG levels, coagulation abnormalities, positive anticardiolipin antibodies and splenomegaly, and sputum fungal cultures indicated Candida albicans infection. Chest CT scans on October 29 showed scattered inflammation in bilateral lungs with cavity formation in the right lower lobe. On October 31, bone marrow punctures displayed one megakaryocyte and widespread platelet distribution in blood marrow smears, and Leishmania amastigotes (Leishman-Donovan bodies) were observed across whole bone marrow smears, prompting patient’s transfer to the Department of Infectious Diseases. On November 5, serum samples were sent to Shanxi Provincial Center for Disease Control and Prevention, where the rK39 rapid immunochromatographic test yielded a positive result. On November 6, metagenomic next-generation sequencing (mNGS) of bone marrows detected 66 596 Leishmania species DNA sequences, including 1 668 L. infantum sequences with relative abundance of 52.02%. Symptomatic treatments were therefore given, including continuous oxygen therapy, transfusion of leukoreduced red blood cells, anti-infection therapy, fluid replacement, correction of hypoalbuminemia, and hemostasis. On November 4, amphotericin B cholesterol sulfate complex was administered intravenously at 50 mg via an intravenous pump, and the dose was adjusted to 100 mg on November 5 and further escalated to the 150 mg (full dose) on November 6. The patient was discharged after the discomfort symptoms gradually subsided. A follow-up examination on December 9 showed excellent antiprotozoal efficacy, with no Leishmania amastigotes detected and the pulmonary cavities disappeared. Subsequent follow-ups indicated no recurrence in the patient.

A case of amoebic proctitis misdiagnosed as ulcerative proctitis

ZHENG Huazhen, WANG Ziqi, JIANG Hongye, WU Xiaodan, ZHENG Huijie

2025, 43(6):  877-880.  doi:10.12140/j.issn.1000-7423.2025.06.021

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This article reports a case of a 59-year-old female patient presenting with hematochezia and lower abdominal pain. Colonoscopy revealed rectal ulcers, and histopathology showed acute inflammation, leading to an initial diagnosis of ulcerative proctitis. After one month of standard anti-inflammatory therapy without clinical improvement, the diagnosis was revised to amoebic proctitis through stool microscopy combined with targeted next-generation sequencing (tNGS). The patient showed rapid symptomatic relief following anti-amebic treatment. This case highlights the diagnostic challenges of Entamoeba histolytica proctitis, including nonspecific clinical manifestations, endoscopic and pathological findings lacking specificity, and the limited sensitivity of conventional pathogen detection methods. For patients with treatment-refractory intestinal ulcers, molecular diagnostic techniques should be actively employed to improve diagnostic accuracy and prevent misdiagnosis and inappropriate treatment.