Since the initiation of the central government fiscal transfer payment program for echinococcosis control in 2005, great strides have been achieved in echinococcosis control in China, progressing from currently basically controlling the transmission of echinococcosis to infection control and elimination. This article analyzes the recent epidemiology of echinococcosis and challenges in the national echinococcosis control program, and proposes future disease control priorities, so as to provide insights into optimization of the echinococcosis control strategy and achieving the goal of infection control and elimination of echinococcosis in China.

Objective To understand the progress, summarize the experiences, and investigate the problems of the national echinococcosis control programme in China in 2023, in order to provide insights for the formulation of echinococcosis control strategies and interventions. Methods All data pertaining to the national echinococcosis control programme across all endemic foci in China in 2023 were captured from the China Information Management System for Prevention and Control of Parasitic Diseases, which was operated by Chinese Center for Disease Control and Prevention, and were used to create a database. The examinations and treatments of echinococcosis in humans, the prevalence of Echinococcus infections in sources of infections, and the prevalence of echinococcosis in intermediate hosts in echinococcosis-endemic areas of China were analyzed using Microsoft Excel 2016. Results Echinococcosis was endemic in 29 415 villages across 3 580 townships from 370 counties (cities, districts, banners) in China by the end of 2023. A total of 25 362 echinococcosis cases were documented in China in 2023, with a prevalence of 0.05% (25 362/46 201 537), and there were 15 878 cases with cystic echinococcosis (62.61%), 7 604 cases with alveolar echinococcosis (29.98%), 300 cases with mixed infections (1.18%) and 1 580 cases with unclassified type (6.23%). There were 1 695 cases with newly diagnosed echinococcosis, including 1 373 cases with cystic echinococcosis (81.00%), 135 cases with alveolar echinococcosis (7.96%), 4 case with mixed infections (0.24%) and 135 cases with unclassified type (10.80%), and there were 177 cases under 12 years of age (10.44%), and 1 518 cases at ages of 12 years and older (89.56%). A total of 5 241 196 person-times received abdominal ultrasound scans for screening of echinococcosis in China in 2023, including 904 370 person-times at age of less than 12 years, and 4 336 826 person-times at ages of 12 years and older, and serological tests were performed among 12 426 person-times suspected of echinococcosis as revealed by ultrasound scans. The detection of echinococcosis by ultrasound screening was 0.02% (72/315 612) among residents under 12 years of age in 370 surveillance sites across China in 2023, including 27 cases with newly diagnosed echinococcosis (37.50%), and the detection of echinococcosis by ultrasound screening was 0.24% (693/288 437) among residents at ages of 12 years and older in surveillance sites with class Ⅰ and Ⅱ endemic counties (cities, districts, banners), including 80 cases with newly diagnosed echinococcosis (11.54%). A total of 17 629 cases with echinococcosis received drug treatment in China in 2023, and 1 920 cases received surgical treatment, including 1 350 cases with cystic echinococcosis (70.31%) and 410 cases with alveolar echinococcosis (21.35%). Follow-up results showed a cure among 2 706 cases with echinococcosis, a response to treatment among 18 729 cases, a failure in treatment among 4 860 cases, deaths among 480 cases (not deaths of echinococcosis), exclusion among 414 cases, loss to follow-up among 273 cases, a failure in follow-up among 138 cases (not reaching the time of follow-up), and migration to other places among 156 cases in China in 2023. A total of 2 189 335 dogs were recorded in echinococcosis-endemic townships in China in 2023, including 2 045 374 dogs registered for management, and dog deworming was performed in 34 561 villages, including deworming with chemotherapeutic agents given to 22 124 218 dog-times and 235 862 doses of chemotherapeutic agents given to wild canines. A total of 395 885 fecal samples were collected from domestic dogs in China in 2023 and 1 968 samples tested positive for Echinococcus coproantigen, with a positive rate of 0.50%, while 75 693 fecal samples were collected from wild canines, and 2 011 samples were tested positive for Echinococcus coproantigen, with a positive rate of 2.66%. A total of 171 102 slaughtered livestock were randomly detected for echinococcosis in China in 2023, and there were 1 533 livestock detected with echinococcosis, with a prevalence rate of 0.90%. In addition, a total of 47 527 wild rodents were examined for echinococcosis in China in 2023, and there were 229 rodents detected with echinococcosis, with a prevalence rate of 0.48%. Conclusion The epidemic status was almost controlled and tended to be stable in China in 2023; however, there are still multiple difficulties and challenges in the national echinococcosis control programme because of the complex factors affecting the transmission of echinococcosis. It is recommended to intensify continuously the echinococcosis control measures, improve the echinococcosis surveillance system, fully exert the role of comprehensive echinococcosis control intervention areas and regional joint prevention and control mechanisms, and promote the implementation of comprehensive echinococcosis control measures.

Objective To identity the dynamic changes of echinococcosis in Sichuan Province, and to provide reference for further prevention and control. Methods According to the National Echinococcosis Disease Surveillance Plan (2020 Edition), surveillance work was conducted in 35 different classified echinococcosis-endemic counties in Sichuan Province. In counties of types Ⅰ and Ⅱ, random cluster sampling method was used to select surveillance villages. Ultrasonographic examination was carried out to screen no less than 1 000 people. In type Ⅲ counties, all outpatients of the ultrasound department in the county-level hospitals were included in the surveillance of echinococcosis. One to five primary schools were randomly selected in each county to conduct student ultrasonographic screening (500 students in counties of types Ⅰ and Ⅱ, 1 500 students in type Ⅲ counties). All fecal samples detected by immunological method, including 10 domestic dog feces of each village in endemic towns and 200 canine feces around settlements or on both sides of rural roads. Each county was required to monitor 50-300 yaks or 100-500 sheep in centralized or scattered slaughterhouses, while the counties endemic with alveolar echinococcosis were required to capture 500 small mammals in mixed endemic counties. And the echinococcosis infections in viscera of intermediate hosts were identified by touching detection or autopsy methods. Questionnaires were used to investigate 300 students’ knowledge and health behavior development regarding to echinococcosis control in each county. Chi-square test was used to compare the rates. Results From 2021 to 2023, a total of 170 881 people were screened in surveillance villages of Sichuan Province, with a total prevalence rate of 0.328% (561/170 881). The prevalence rates from 2021 to 2023 were 0.334% (209/62 639), 0.458% (257/56 103), and 0.182% (95/52 139), respectively, with statistically significant differences (χ² = 62.94, P < 0.01). The detection rates of new patients from 2021 to 2023 were 3.19/100 000, 0, and 1.92/100 000, respectively, with no statistically significant difference (Fisher value = 1.62, P > 0.05). 77 308 primary school students were screened from 2021 to 2023, and no new patients with echinococcosis were found. The prevalence rate of alveolar echinococcosis (0.181%, 310/170 881) was higher than that of cystic echinococcosis (0.132%, 226/170 881) (χ² = 13.19, P < 0.01), and the prevalence rate of female (0.395%, 360/91 102) was higher than that of male (0.252%, 210/79 779) (χ² = 22.66, P < 0.01). There were statistical differences in the prevalence among different age groups and regions (χ² = 77.74, 261.54, P < 0.01). The positive rates of Echinococcus spp. coproantigen in domestic dogs were 0.50% (133/26 450), 0.38% (108/28 264) and 0.24% (72/29 847) from 2021 to 2023, showing a decreasing trend by year (χ² = 26.19, P < 0.01). The positive rate of canine feces in the wild in 2021 (1.60%, 37/2 312) was significantly higher than it in 2022 (0.40%, 11/2 720) and 2023 (0.35%, 10/2 844) (χ² = 33.47, P < 0.01). During 2021 to 2023, the prevalence in livestock were 0.92% (82/8 898), 1.07% (78/7 312) and 1.24% (92/7 416), respectively, with no statistical difference between years (χ² = 3.90, P > 0.05), and the prevalence in small mammals were 3.48% (442/12 684), 2.58% (231/8 955) and 1.39% (129/9 286), with a decreasing trend by year (χ² = 93.20, P < 0.01). The awareness rates of knowledge on echinococcosis prevention among primary school students from 2021 to 2023 were 92.62% (11 114/11 999), 92.17% (10 221/11 089) and 93.32% (10 491/11 242), respectively, with statistical significance (χ² = 11.05, P < 0.05). From 2021 to 2023, the qualified rates of no touching the dog was 75.47% (9 056/11 999), 73.24% (8 122/11 089) and 82.19% (9 240/11 242), respectively, and that of washing hands before meals were 70.72% (8 486/11 999), 75.63% (8 387/11 089) and 80.76% (9 079/11 242), respectively, and that of no feeding dogs with raw livestock viscus were 87.82% (10 537/11 999), 83.25% (9 232/11 089) and 92.47% (10 395/11 242). The difference in health behavior qualification rates of students between different years was statistically significant (χ² = 274.81, 316.96, 444.35; all P < 0.01). Conclusion The detection rate of new echinococcosis patients and the positive rate of animal hosts in Sichuan have decreased steadily or maintained a low level, and the rate of health awareness and behavior among the population is gradually increasing. However, some regions have significant fluctuations, and it is necessary to continuously strengthen comprehensive prevention and control measures to prevent the resurgence of the endemic.

Objective To evaluate the effectiveness of comprehensive echinococcosis control measures in Yunnan Province from 2017 to 2023, so as to provide insights into formulation of the echinococcosis control strategy in the province in the future. Methods The central government fiscal transfer payment program for echinococcosis prevention and control was implemented across 24 counties (cities, districts) in 9 prefectures (cities) of Yunnan Province in 2017, and establishment of the echinococcosis surveillance and laboratory testing system, standardized management of echinococcosis patients, case investigation, investigation and disposal of foci, management of dogs and intermediate host animals and health education were performed in echinococcosis-endemic areas. The comprehensive echinococcosis control measures, echinococcosis control reports and echinococcosis surveillance reports were collected in Yunnan Province from 2016 to 2023, and the prevalence of human echinococcosis, positive rate of Echinococcus coproantigen in dogs, prevalence of echinococcosis in livestock, and awareness of echinococcosis control knowledge among residents were estimated. All analyses were performed with the statistical software SPSS 21.0, and the trends were tested with chi-square test. In addition, the relative risk was calculated from 2017 to 2023 based on the 2016 data, in order to evaluate the effectiveness of comprehensive echinococcosis control measures in Yunnan Province. Results An echinococcosis control workstation and 2 province-level echinococcosis testing reference laboratories were built in Yunnan Province during the period between 2017 and 2023, when B-mode ultrasonography screening for human echinococcosis among 1.83 million person-times, 365 times of follow-up of echinococcosis patients, 254 case investigations, a 100% proportion of standardized case management and investigation and disposal of 5 foci were conducted. The prevalence of human echinococcosis reduced from 38.17/105 (24/62 874) at baseline to 1.42/105 (11/774 609) in 2023, with a reduction of 96.28% (χ² = 187.881, P < 0.05). The awareness of echinococcosis control knowledge increased from 43.50% (5 075/11 668) among residents at baseline to 89.20% (6 680/7 489) in 2023 (χ² = 922.835，P < 0.05), while the awareness of echinococcosis control knowledge increased from 77.86% (2 687/3 451) among primary school students in 2019 to 95.21% (7 097/7 454) in 2023 (χ² = 44.170，P < 0.05). The positive rate of Echinococcus coproantigen reduced from 4.38% (186/4 246) in dogs in 2017 to 1.68% (141/8 392) in 2023 (χ² = 367.928，P < 0.05), and the prevalence of echinococcosis reduced from 0.23% (8/3 472) in livestock at baseline to 0.12% (7/5 663) in 2023 (χ² = 1.492，P > 0.05). In addition, the risk of human echinococcosis reduced by 97.99% in 2023 relative to in 2017, and the risk of dog-associated transmission of echinococcosis reduced by 90.76% in 2023 relative to in 2018. Nevertheless, there was no significant difference in the risk ratio of the prevalence of echinococcosis in livestock among years (P > 0.05). Conclusion Remarkable achievements had been obtained in the comprehensive echinococcosis control measures in Yunnan Province from 2017 to 2023; however, transmission of echinococcosis remained among animal sources of infections. Intensified management of animal sources of infections is required to effective block the transmission chain of echinococcosis.

Objective To investigate the epidemiology of echinococcosis in Inner Mongolia Autonomous Region, so as to provide insights into echinococcosis control. Methods Baseline surveys on the epidemiology of echinococcosis were performed using a stratified cluster sampling method in 36 banners (counties, cities, districts) across 11 leagues (cities) in Inner Mongolia Autonomous Region from 2012 to 2018. Approximately 3 200 residents were recruited in each banner (county, city, district), and human echinococcosis was diagnosed with ultrasound. Fecal samples were randomly collected from one dog per household in the surveyed villages, and the Echinococcus coproantigen was detected using ELISA in dogs. In addition, Echinococcus infection was identified using necropsy in livestock (sheep). Results Echinococcosis patients were diagnosed in 22 banners (counties, cities, districts), including 15 in pastoral areas, 5 in semi-agricultural and semi-pastoral areas, and 2 in agricultural areas. The highest number of echinococcosis cases was recorded in Xiwuzhumuqin Banner (16 cases, 21.6% of all cases), and there were 6 banners (counties, cities, districts) with echinococcosis prevalence of 0.10% to 0.50%, and 16 with prevalence of 0.03% to 0.10%. A total of 115 155 residents received ultrasound screening for echinococcosis, and 74 residents were diagnosed with echinococcosis (all cystic echinococcosis), with a prevalence rate of 0.11% (74/115 155). Women (0.08% prevalence, 51/60 544), residents at ages of 30 to 59 years (accounting for 60.81%, 45/74), herders (accounting for 55.41%, 41/74), residents with an educational level of primary school (0.10% prevalence, 39/39 237), and nomadic populations (0.29% prevalence, 6/2 097) were identified at a high risk of echinococcosis. A total of 17 909 dog feces samples were collected from 36 banners (counties, cities, districts), and the positive rate of Echinococcus coproantigen was 1.84% (330/17 909) in dogs. Dog feces positive for Echinococcus coproantigen was detected in 26 banners (counties, cities, districts), and there was one banner with a 5.00% and higher positive rate of Echinococcus coproantigen in dogs (New Barag Right Banner, 11.75%), 18 banners (counties, cities, districts) with positive rates of 1.00% to 5.00% and 8 banners (counties, cities, districts) with positive rates of 0.29% to 1.00%. A total of 32 100 sheep were examined for echinococcosis in 33 banners (counties, cities, districts), and the prevalence of echinococcosis was 0.46%. Sheep with echinococcosis were found in 14 banners (counties, cities, districts), with echinococcosis prevalence of 0.10% to 7.30%, and there were 3 banners (counties, cities, districts) with echinococcosis prevalence of > 1.00%, all in pastoral areas. Conclusion Echinococcosis is widely but overall lowly prevalent in Inner Mongolia, and there are obvious regional and population clustering distributions. Intensified echinococcosis prevention and control is recommended in key areas and populations.

Objective To explore the method, frequency and dose of deworming in stray dogs and wild canine, evaluate the deworming effect, and provide reference for controlling hydatid infection in stray dogs and wild canine. Methods Four villages were randomly selected in each of the 3 alveolar echinococcosis endemic areas in Ningxia (Xiji, Yuanzhou and Haiyuan) as deworming sites for stray dogs, and 2 sites were selected in each of the natural forest farms in the 3 counties (districts) as deworming sites for wild canine. The praziquantel was administered from April to June and September in 2023 (chewable tablets, 0.1 g/tablet). The linear method and circular method were used in both stray dog deworming sites and wild canine deworming sites, and each administration method was performed with dosages of 1-tablet and 2-tablet. Three days after administration, the drug swallow rate at each deworming site was calculated. Fresh fecal samples were collected from canine, and Echinococcus coproantigen was detected by ELISA to calculate the positive rate. The cloth clip method was used to capture The small rodents at wild canine deworming sites were captured using trap layout method and identified. The suspected echinococcosis lesions of small rodents were paraffin sectioned and stained with hematoxylin and eosin. The protoscoleces were observed under the microscope to calculate the rodent prevalence rate. The database was established using Epi info 3.5.3 software and SPSS 22.0 software was used for statistical analysis. Rates were compared by ANOVA, chi-square test or U test. Results The drug swallow rate at the stray dog deworming sites using the circular method was 70.98% (269/379), which was higher than that of 45.38% (172/379) using the linear method (F = 13.577, P < 0.05); there were no statistically significant differences in the swallow rates between the linear method and the circular method for different dosages (F = 0.731, 1.124, both P > 0.05). The drug swallow rate at the wild canine deworming sites using the linear method was 72.94% (124/170), which was higher than that of 49.12% (84/171) using the circular method (F = 4.950, P < 0.05); there were no statistically significant differences in the swallow rates between the linear method and the circular method for different dosages (F = 0.200, 0.341, both P > 0.05). The coproantigen positivity rate of stray dog deworming sites using the circular method was 2.84% (14/493), which was lower than that of 5.66% (28/495) using the linear method (χ² = 4.423, P < 0.05); and the coproantigen positivity rate of wild canine deworming sites using the linear method was 2.85% (7/246), which was lower than that of 6.94% (17/245) using the circular method (χ² = 4.013, P < 0.05). A total of 562 small rodents of 9 species were captured, with a prevalence rate of 0.89% (5/562). All 5 diseased rodents were Spermophilcus alashanicus, with the prevalence rate of 1.58% (5/317). The prevalence rates of small rodents at the deworming sites of the linear method and the circular method were 1.18% (2/169) and 2.03% (3/148), respectively, with no statistically significant difference (χ² = 0.350, P > 0.05). Conclusion The linear method is suitable for administering drugs to wild canine along highways, streams and other environments. The circular method is suitable for administering drugs to stray dogs in living environments. Continuous administration for more than 3 months can effectively control hydatid infection in canine hosts.

Objective To construct a method for evaluating the risk of transmission of echinococcosis endemic site. Methods By analyzing the characteristics of the echinococcosis endemic in Sichuan Province from 2019 to 2023 and conducting in-depth interviews with experts, the items of assessment system of echinococcosis endemic site were determined and the first draft of the risk assessment system of echinococcosis endemic site was formed. The final draft of the risk assessment system for transmission of echinococcosis endemic site was formed through expert inquiry. Selected villages in Sichuan Province with newly diagnosed patients and patients aged 15 and above in 2023 to conduct transmission risk assessment. Intervened for one year based on the risk level of the endemic site, compared the risk level before and after intervention, and verify the effectiveness of the assessment system. Results A total of 11 experts participated in the questionnaire inquiry, and the effective response rates for both rounds of questionnaires were 11/11. The risk assessment system of echinococcosis endemic site was constructed successfully, including the evaluation scale of transmission risk of echinococcosis (4 first-grade indexes, 15 second-grade indexes, 39 third-grade indexes), the geographical scope of intervention for transmission risk of echinococcosis and the time range table for monitoring and intervention of transmission risk of echinococcosis. What’s more, a risk assessment scale for the transmission of echinococcosis endemic site was developed, and the criteria for risk levels were determined: general risk with a score below 36, high risk with a score between 36 and 55, and maximum high risk with a score above 55. The verification results of four endemic sites, namely Rangtang County endemic site, Ruoergai County endemic site 1, Ruoergai County endemic site 2 and Litang County endemic site, showed that the risk level of each endemic point remained unchanged or decreased after intervention, confirming the effectiveness of the evaluation system. Conclusion The risk assessment system of echinococcosis endemic site constructed in this study is reliable and practical, and can provide reference for the disposal and intervention of echinococcosis endemic site.

Objective To analyze the cost-effectiveness of population screening and patient treatment for echinococcosis in Xizang from 2015 to 2022, so as to provide insights into formulation or adjustment of the echinococcosis control strategies and measures. Methods The costs for population echinococcosis screening and echinococcosis patient treatment, newly diagnosed echinococcosis cases, and treatment efficiency in Xizang from 2015 to 2022 were collected from the annual report system of the Hydatid Disease Prevention and Control Program in Xizang Autonomous Region, and the cost-effectiveness of echinococcosis screening and treatment was calculated in each year from 2015 to 2022 and in each prefecture/city. The incremental cost-effectiveness atio (ICER) was used to examine the association of the difference (∆E) between the actual disability-adjusted life years (DALYs) due to echinococcosis and the estimated DALYs without interventions with the total costs of echinococcosis screening and treatment (∆C) in Xizang, and the willingness-to-pay (WTP) threshold was measured as a ratio of ICER to per capita gross domestic product (GDP). The difference in the overall response to treatment was tested for statistical significance with Kruskal-Wallis (K-W) test among years, followed by pairwise comparisons with Bonferroni corrections. Results The total costs of population screening for echinococcosis were 54.171 million yuan in Xizang from 2015 to 2022, with the highest in 2017 (27.624 million yuan). The cost-effectiveness ratio for screening newly detected echinococcosis cases appeared a tendency towards a rise over years from 2017 to 2022, from 1 641 yuan/person in 2017 to 21 131 yuan/person in 2022. The proportion of newly detected echinococcosis cases by screening was highest in Shigatse City (0.10%) and lowest in Shannan City (0.00%) in 2022, and the cost-effectiveness ratios were 10 000 yuan/person and 209 000 yuan/person, respectively. The total costs for echinococcosis patient treatment were 80.560 million yuan in Xizang from 2015 to 2022, peaking in 2018 (39.055 million yuan), including 58.807 million yuan for surgical treatment (73.0%) and 21.754 million yuan for drug treatment (27.0%). The overall cure rate of echinococcosis patients was 11.1% in Xizang from 2015 to 2022, with no significant annual trends in cure rates; however, the proportion of improvements appeared a tendency towards a rise during the study period, with 68.6% in 2022. The actual DALYs due to echinococcosis were lower than the estimated DALYs without interventions in Xizang, and the averted DALYs loss increased from 6 293.03 in 2017 to 11 487.84 in 2022. The average ICER for population echinococcosis screening and patient treatment was 11 460.3 yuan/year in Xizang from 2017 to 2022, with a WTP of 0.196. Conclusion Xizang has given large financial investments and achieved a satisfactory efficiency for population screening and patient treatment of echinococcosis from 2017 to 2022, with a high fund utilization efficiency. Through comprehensive screening, a large number of early-stage echinococcosis patients were identified, and surgical treatments and medication were actively arranged. However, regions with currently low screening efficiency are no longer suitable for large-scale population screening. It is recommended to adjust screening strategies and methods according to local conditions. For the treatment of echinococcosis patients, it is advised to enhance surgical training to improve surgical standards and outcomes, strengthen the standardized management of patients undergoing drug treatment, and improve medication adherence, thereby further enhancing the treatment effectiveness for echinococcosis patients.

Objective To explore the cytotoxic effect of mouse immune cells on the Echinococcus granulosus protoscolex and understand the types of immune cells involved and the cytokines secretion changes. Methods Healthy C57BL/6 mice were used to extract splenocytes and peritoneal macrophages. The protoscoleces from sheep E. granulosus cysts were collected and grouped (3 000 per group). To select the immune cell type exhibiting a stronger inhibitory effect on the protoscoleces, macrophage co-culture group and splenocyte co-culture group were co-cultured with 6 × 10⁶ macrophages or splenocytes, respectively. To choose an optimal cell count, co-culture group 1-5 were co-cultured with 1.2 × 10⁶, 2.4 × 10⁶, 4.8 × 10⁶, 7.2 × 10⁶ and 9.6 × 10⁶ immune cells, respectively. The co-culture system was established. The E. granulosus cyst fluid and tumor necrosis factor-α (TNF-α) inhibitor were added into the co-culture system respectively, and the activity of protoscoleces and the changes in concentration of TNF-α, interleukin 6 (IL-6), IL-10 and transforming growth factor-β (TGF-β) in the co-culture supernatant were observed. Eosin staining was used to detect the activity of protoscoleces, the dichlorodihydrofluorescein diacetate (DCFH-DA) method was used to measure the of reactive oxygen species levels, the JC-1 method was used to assess mitochondrial membrane potential, Western blotting was performed to detect the expression levels of the Bcl-2 associated X protein (Bax) and cysteinyl aspartate specific proteinase 3 (caspase-3), and ELISA was used to measure the concentration of cytokines in the culture supernatant. Independent samples t-test was used for comparisons between two groups and one-way ANOVA was used for multiple groups. Results On day 6 of co-culture, the protoscoleces activity in the macrophage co-culture group and the splenocyte co-culture group was (25.07 ± 0.40)% and (76.18 ± 0.31)%, respectively. The protoscoleces activity in the macrophage co-culture group was lower than that in the splenocyte co-culture group at all time points (F = 564.20, P < 0.05). On day 4 of co-culture, the relative fluorescent intensity of reactive oxygen species in the protoscoleces of the macrophage co-culture group was 32.20 ± 7.85, which was higher than that of the splenocyte co-culture group (12.44 ± 2.93) (t = 7.07, P < 0.05). On day 6 of co-culture, the relative expression levels of caspase-3 protein in the macrophage co-culture group and the splenocyte co-culture group were 1.28 ± 0.02 and 1.16 ± 0.02, respectively, and the relative expression levels of Bax protein were 1.29 ± 0.01 and 0.46 ± 0.01, respectively. The relative expression levels of caspase-3 and Bax protein in the macrophage co-culture group were higher than those in the splenocyte co-culture group at all time points (F = 55.87, 167.20; both P < 0.05). Macrophages exhibited a stronger inhibitory effect on protoscoleces than splenocytes. On day 6 of co-culture, the relative fluorescent intensities of mitochondrial membrane potential in protoscoleces from co-culture groups 1-5 were 20.15 ± 8.96, 24.40 ± 9.71, 48.41 ± 10.20, 94.62 ± 8.72 and 112.85 ± 24.23, respectively, all of which were higher than that of the protoscolex control group (2.50 ± 1.02) (F = 26.18, P < 0.01). On day 6 of co-culture, the relative expression levels of caspase-3 protein in protoscoleces from co-culture group 4 and 5 were 1.35 ± 0.03 and 1.49 ± 0.05, respectively, which were higher than that of the protoscolex control group (0.28 ± 0.01) (t = 17.03, 10.60; both P < 0.05). The relative expression levels of Bax protein in protoscoleces from co-culture groups 4 and 5 were 1.34 ± 0.01 and 1.38 ± 0.04, respectively, which were higher than that of the protoscolex control group (0.78 ± 0.04) (t = 6.68, 6.46; both P < 0.05). On day 4 of co-culture, the concentrations of TNF-α, IL-6 and TGF-β in the supernatants of co-culture group 4 were (240.90 ± 17.29), (435.90 ± 12.33) and (137.10 ± 6.62) pg/ml, respectively, all of which were higher than those in the cell control group [(42.02 ± 0.52), (65.72 ± 1.91), (24.72 ± 1.78) pg/ml] (t = 54.52, 15.97, 17.59; all P < 0.05). The concentration of IL-10 in the supernatant of co-culture group 4 was (42.16 ± 1.45) pg/ml, which was not significantly different from that of the cell control group [(45.64 ± 1.03) pg/ml] (t = 1.29, P > 0.05). The concentrations of TNF-α, IL-6, IL-10 and TGF-β in the supernatants of the co-culture groups at all time points were higher than those in the cell control group (F = 294.66, 450.50, 687.72, 660.15; all P < 0.05). On days 1, 3, 5 and 7 of co-culture, the relative fluorescent intensities of mitochondrial membrane potential in protoscoleces from the cyst fluid group were 4.46 ± 1.25, 4.33 ± 0.39, 4.89 ± 0.77 and 7.97 ± 0.62, respectively, all of which were lower than those in the macrophage group (5.67 ± 1.72, 13.60 ± 0.50, 35.28 ± 5.65, 77.50 ± 9.60) (F = 115.90, P < 0.01). On day 6 of co-culture, the concentrations of TNF-α, IL-6, IL-10 and TGF-β in the supernatant of the cyst fluid group were (64.12 ± 2.65), (1 049.65 ± 25.70), (230.30 ± 12.98) and (138.57 ± 13.71) pg/ml, respectively, and those in the macrophage group were (41.61 ± 1.31), (68.00 ± 0.42), (56.15 ± 6.43) and (32.94 ± 4.90) pg/ml, respectively. The concentrations of TNF-α, IL-6, IL-10 and TGF-β in the supernatants of the cyst fluid group at all time points were higher than those in the macrophage group (F = 289.80, 366.50, 145.40, 32.94; all P < 0.05). On day 7 of co-culture, the protoscoleces activity in the macrophage group and the inhibitor group was (21.18 ± 1.61)% and (94.31 ± 2.58)%, respectively. The protoscoleces activity in the inhibitor group was higher than that in the macrophage group at all time points (F = 1 810.00, P < 0.05). On days 2, 4 and 6 of co-culture, the concentrations of TNF-α in the inhibitor group were (33.55 ± 7.48), (13.78 ± 4.96) and (19.20 ± 0.69) pg/ml, respectively, all of which were lower than those in the macrophage group [(209.24 ± 9.90), (209.47 ± 10.55), (211.36 ± 13.66) pg/ml] (t = 33.16, 30.46, 23.76; all P < 0.05). Conclusion Macrophages co-cultured in vitro with the E. granulosus protoscoleces could express cytokines such as TNF-α, which inhibits the activity of the protoscoleces and promotes their apoptosis. Cyst fluid from E. granulosus and TNF-α inhibitors could reduce the secretion of TNF-α by macrophages, thereby alleviating the killing effect of macrophages on the protoscoleces.

Objective To establish a rapid and convenient assay for detection of Echinococcus and identification of E. granulosus based on recombinant polymerase amplification (RPA). Methods The Echinococcus cytochrome C oxidase subunit 1 (co1) and NADH dehydrogenase subunit 5 (nd5) of genes were selected as target sequences, and specific primers and probes were designed with the software Primer Premier 6 and screened to establish a dual-probe fluorescent RPA assay for detection of Echinococcus nucleic acid. The sensitivity of the dual-probe fluorescent RPA assay was evaluated with co1 and nd5 recombinant plasmid DNA at concentrations of 10⁴, 10³, 10², 10, and 1 copies/μl, and the specificity of the assay was evaluated with genomic DNA from E. multilocularis, Taenia solium, Schistosoma japonicum, Necator americanus, Nippostrongylus brasiliensis, Clonorchis sinensis, Giardia lamblia, Cryptosporidium parvum, Toxoplasma gondii and Babesia microti. The feasibility of the method was evaluated using 23 simulated samples (prepared by mixing E. granulosus protoscolex DNA with DNA extracted from negative canine feces) and 5 field-collected dog fecal samples positive for E. granulosus (confirmed by PCR). Results The optimal reaction procedure of the FAM/HEX dual-probe fluorescence RPA assay for detection of Echinococcus genomic DNA was oscillation in a metal bath at 39 ℃, 300 r/min for 4 minutes, followed by detection of the fluorescence signal for 12.5 minutes using real-time quantitative PCR (qPCR) assay. The total reaction time of performance was 16.5 minutes. The minimum detection limits of the dual-probe fluorescent RPA assay were 10 copies/μl for the co1 recombinant plasmid and 100 copies/μl for the nd5 recombinant plasmid, respectively, and the assay yielded the FAM/HEX dual fluorescence signals for E. granulosus, FAM single fluorescence signal for E. multilocularis, and no specific reaction to the genomic DNA from T. solium, S. japonicum, N. americanus, N. brasiliensis, C. sinensis, G. lamblia, C. parvum, T. gondii and B. microti. In simulated DNA samples, FAM/HEX double positive fluorescence signals were generated in 16 samples containing E. granulosus genomic DNA, including 8 simulated samples containing E. granulosus genomic DNA alone and 8 samples containing both E. granulosus and E. multilocularis genomic DNA, and the FAM single positive fluorescence signal was produced in 7 samples containing E. multilocularis genomic DNA alone. In addition, FAM/HEX double positive fluorescence signals were found in 5 field captured dog fecal samples positive for E. granulosus genomic DNA. No positive signals were observed in other DNA samples. The detection results of the dual-probe fluorescent RPA assay were consistent with those by PCR assay. Conclusion A dual-probe fluorescent RPA assay has been successfully developed for detection of Echinococcus and distinguishing E. granulosus, which is low in the detection limit, high in specificity, and simple and rapid in procedures.

Objective To explore the effects and mechanisms of Echinococcus granulosus cyst fluid (CF) on the activation and migration of hepatic stellate cells (HSC). Methods Liver lesion tissues of cystic echinococcosis (CE) patients were obtained, and the fibrosis of liver tissues was observed using hematoxylin and eosin (HE) staining and Masson staining. HSCs were cultured in vitro for 48 and 72 hours in culture medium without CF and with 10%, 20% and 40% CF, respectively. Western blotting was used to detect the relative protein expression levels of α-mooth muscle actin (α-SMA), collagen type I alpha 1 (Col1a1), phosphatidylinositol 3 kinase (PI3K), phosphorylated PI3K (p-PI3K), protein kinase B (Akt) and phosphorylated Akt (p-Akt). Real-time quantitative PCR (qPCR) was used to detect the relative mRNA transcription levels of α-SMA, Col1a1 and Col3a1. Scratch assay and Transwell assays were conducted to assess the effects of CF on HSC migration. Results HE staining showed that the outer capsule of E. granulosus in the liver lesion tissues of CE patients was transparent and homogeneous with fibrous strips, and Masson staining showed a large amount of collagen fiber deposition in the outer capsule. Western blotting results showed that after 72 hours of intervention, the relative protein expression levels of α-SMA in the control group, 10% CF group, 20% CF group and 40% CF group were 0.359 ± 0.043, 0.641 ± 0.088, 0.900 ± 0.084 and 1.111 ± 0.027, respectively. The 10% CF group, 20% CF group, and 40% CF group had higher levels than the control group (t = 5.236, 10.050, 13.980; all P < 0.05). The relative expression levels of Col1a1 in each group were 0.392 ± 0.057, 0.546 ± 0.022, 0.854 ± 0.034 and 1.127 ± 0.057, respectively. The 10% CF group, 20% CF group and 40% CF group had higher levels than the control group (t = 4.228, 12.630, 20.090; all P < 0.05). qPCR results showed that after 72 hours of intervention, the relative mRNA transcription levels of α-SMA in the control group, 10% CF group, 20% CF group and 40% CF group were 1.000 ± 0.093, 1.437 ± 0.106, 1.453 ± 0.105 and 1.697 ± 0.154, respectively. The 10% CF group, 20% CF group and 40% CF group had higher levels than the control group (t = 4.604, 4.769, 7.339; all P < 0.05). The relative mRNA transcription levels of Col1a1 in each group were 0.856 ± 0.042, 1.067 ± 0.049, 1.283 ± 0.128 and 1.582 ± 0.046, respectively. The 20% CF group and 40% CF group had higher levels than the control group (t = 6.932, 11.790; both P < 0.05). The relative mRNA transcription levels of Col3a1 in each group were 0.611 ± 0.054, 0.908 ± 0.041, 1.000 ± 0.045 and 1.239 ± 0.101, respectively. The 10% CF group, 20% CF group and 40% CF group had higher levels than the control group (t = 5.599, 7.346, 11.850; all P < 0.05). Western blotting results showed that the relative protein expression levels of p-PI3K/PI3K in the control group, 10% CF group, 20% CF group and 40% CF group were 0.346 ± 0.050, 0.716 ± 0.054, 0.941 ± 0.114 and 1.276 ± 0.004, respectively. The 10% CF group, 20% CF group, and 40% CF group had higher levels than the control group (t = 6.669, 10.740, 16.780; all P < 0.01). The relative protein expression levels of p-Akt/Akt in each group were 0.524 ± 0.111, 0.815 ± 0.019, 1.043 ± 0.052 and 1.333 ± 0.054, respectively. The 10% CF group, 20% CF group and 40% CF group had higher levels than the control group (t = 5.268, 9.413, 14.670; all P < 0.01). Scratch assay results showed that after 24 hours of intervention, the cell migration rates in the 10% CF group, 20% CF group and 40% CF group were (73.6 ± 2.1)%, (88.2 ± 2.1)% and (96.5 ± 1.2)%, respectively, all higher than (55.1 ± 2.9)% in the control group (t = 10.510, 18.820, 23.540; all P < 0.05). Transwell assay results showed that after 24 hours of intervention, the average number of cells per field in the control group, 10% CF group, 20% CF group and 40% CF group were (62.333 ± 8.145), (82.333 ± 8.505), (132.333 ± 17.620) and (164.333 ± 11.060), respectively. The 20% CF group and 40% CF group had higher numbers than the control group (t = 7.173, 10.450; both P < 0.01). Conclusion E. granulosus CF could promote the activation of HSC and enhance their migration ability through the PI3K/Akt pathway.

Objective To understand the changes in the migration and differentiation of immune cells in the peritoneal cavity of mice infected with Echinococcus multilocularis, and to explore the mechanism of immune function. Methods Spleen tissues of enhanced green fluorescent protein (EGFP) mice were lysed to isolate EGFP-labeled immune cells. C57BL/6J mice were intraperitoneally injected with 2.5 × 10⁷ traced immune cells. Five days later, 3 000 protoscoleces of E. multilocularis were inoculated via the hepatic portal vein. After 19 weeks of infection, the liver and spleen tissues of the infected mice were collected. After frozen sectioning, the sections were stained with 4,6-diamidino-2-phenylindole (DAPI), and immunofluorescence was used to observe the distribution of the traced immune cells. Immune cells from peripheral blood, liver, spleen and peritoneal cavity of infected mice were isolated, and the migration distribution and cell typing of EGFP-labeled immune cells in different tissues were detected by flow cytometry, as well as the secretion of cytokines interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) and interleukin-4 (IL-4). Results The immunofluorescence results showed that the EGFP-traced cells mainly migrated to the spleen tissue of the mice. In the liver tissue, the proportions of various types of traced lymphocytes from high to low were as follows: CD8⁺ T cells (79.63 ± 5.00)%, CD4⁺ T cells (7.08 ± 4.00)%, and B cells (2.01 ± 1.00)% (F = 396.20, P < 0.01). In the spleen tissue, the proportions of various types of lymphocytes from high to low were as follows: CD8⁺ T cells (73.97 ± 3.56)%, CD4⁺ T cells (11.92 ± 5.02)% and B cells (3.86 ± 0.32)% (F = 349.00, P < 0.01). In the peritoneal cavity, the proportions of various types of lymphocytes from high to low were as follows: CD8⁺ T cells (45.27 ± 4.53)%, CD4⁺ T cells (34.60 ± 8.00)% and B cells (15.03 ± 6.38)% (F = 16.90, P < 0.05). The numbers of traced CD4⁺ T cells in the liver, spleen, and peritoneal cavity were 2 700 ± 3 120, 14 407 ± 11 958 and 3 376 ± 1 481 respectively. The numbers of traced CD8⁺ T cells were 26 407 ± 22 190, 109 179 ± 92 692 and 4 245 ± 798 respectively. The numbers of traced B cells were 698 ± 719, 5 794 ± 4 971 and 1 476 ± 840 respectively. All three types of traced lymphocytes were mainly distributed in the spleen tissue (F = 2.51, 3.03, 2.62, all P > 0.05). In the liver tissue, the proportions of various types of myeloid cells from high to low were as follows: dendritic cells (3.94 ± 1.33)%, monocytes (3.79 ± 1.80)%, macrophages (3.13 ± 1.85)%, eosinophils (2.40 ± 1.81)%, myeloid-derived suppressor cells (0.76 ± 0.17)% and neutrophils (0.67 ± 0.04)% (F = 3.21, P < 0.05). In the spleen tissue, the proportions of various types of myeloid cells from high to low were as follows: dendritic cells (1.86 ± 0.89)%, monocytes (0.41 ± 0.09)%, macrophages (0.23 ± 0.03)%, neutrophils and myeloid-derived suppressor cells [both (0.13 ± 0.14)%], and eosinophils (0.04 ± 0.04)% (F = 42.70, P < 0.01). In the peripheral blood, the proportions of various types of myeloid cells from high to low were as follows: dendritic cells (0.37 ± 0.28)%, monocytes (0.22 ± 0.27)%, macrophages (0.22 ± 0.21)%, neutrophils and myeloid-derived suppressor cells [both (0.17 ± 0.08)%], and eosinophils (0.05 ± 0.08)% (F = 0.92, P > 0.05). Among the traced CD4⁺ T cells in the liver tissue, the cells secreting IFN-γ, TNF-α, and IL-4 accounted for (41.17 ± 19.92)%, (30.70 ± 3.35)%, and (2.78 ± 4.81)% respectively (F = 8.22, P < 0.05). Among the traced CD8⁺ T cells, the cells secreting IFN-γ, TNF-α, and IL-4 accounted for (64.17 ± 3.17)%, (24.85 ± 15.66)%, and (5.67 ± 6.35)% respectively (F = 27.08, P < 0.01). Among the traced CD4⁺ T cells in the spleen tissue, the cells secreting IFN-γ, TNF-α, and IL-4 accounted for (52.43 ± 19.43)%, (27.67 ± 18.99)%, and (4.77 ± 2.23)% respectively (F = 6.88, P < 0.05). Among the traced CD8⁺ T cells, the cells secreting TNF-α, IFN-γ, and IL-4 accounted for (40.53 ± 17.79)%, (38.97 ± 18.04)%, and (3.23 ± 3.09)% respectively (F = 6.15, P < 0.05). Among the traced CD4⁺ T cells in the peritoneal cavity, the cells secreting IFN-γ, TNF-α, and IL-4 accounted for (64.33 ± 6.82)%, (46.43 ± 13.44)%, and (4.03 ± 3.51)% respectively (F = 36.04, P < 0.01). Among the traced CD8⁺ T cells, the cells secreting IFN-γ, TNF-α, and IL-4 accounted for (53.57 ± 19.17)%, (21.47 ± 8.54)%, and (6.13 ± 8.63)% respectively (F = 10.24, P < 0.05). Conclusion Immune cells in the peritoneal cavity of E. multilocularis infected mice mainly migrate to the spleen tissue, and the cell types are mainly CD8⁺ T cells and dendritic cells. T cells mainly secreted IFN-γ, which showed Th1 immune response.

Objective To investigate the effects of Echinococcus multilocularis infection on the expression of CD47 and signal regulatory protein α (SIRPα) in macrophages and the function of macrophages, and to provide evidence for exploring the pathogenesis of alveolar echinococcosis (AE). Methods Liver tissue samples were collected from 30 AE patients. Tissues were categorized into close liver tissue (CLT) located 0.5 cm from the lesion and distal liver tissue (DLT) located more than 2 cm from the lesion. Paraffin sections were used for immunohistochemistry and frozen sections were used for immunofluorescence to analyze the expression levels of CD47 and SIRPα in CLT and DLT. Mouse monocyte-macrophage leukemia cells (RAW264.7) were co-cultured with E. multilocularis protoscoleces at a ratio of 500 ∶ 1. Cells were collected before the addition of protoscoleces and after co-culture for 24 and 48 hours, respectively. The expression changes of CD47 and SIRPα in macrophages were detected by flow cytometry, cellular immunofluorescence and quantitative real-time PCR (qPCR). RAW264.7 cells were divided into control group, infection group, inhibitor group and inhibitor-infection group (1 × 10⁵ cells per group). The infection group and inhibitor-infection group were supplemented with protoscoleces (200 per group), while the inhibitor group and inhibitor-infection group were treated with a CD47/SIRPα binding inhibitor (NCGC00138783TFA, 10 µmol/L). Enhanced green fluorescent protein (EGFP)-labeled Escherichia coli were added to each group (1 × 10⁶ per group) after co-culture for 48 hourse. Following fluorescence observation, the phagocytic capacity of macrophages was assessed by flow cytometry. Additionally, the mRNA relative transcription levels of macrophage polarization markers and cytokines were detected by qPCR. Independent samples t-test or paired t-test was used for comparisons between two groups, one-way ANOVA was used for multiple groups, and Tukey’s HSD method was used for multiple comparisons. Results Immunohistochemical analysis revealed that in AE patients, the proportions of CD47 and SIRPα positive cells in CLT were (61.99 ± 3.61)% and (54.06 ± 1.85)%, respectively, which were higher than (57.08 ± 3.38)% and (40.77 ± 1.49)% in DLT (t = 9.434, 58.840; both P < 0.01). Immunofluorescence analysis revealed that the Pearson correlation coefficient between CD47 and SIRPα in CLT was 0.66 ± 0.02, which was higher than that in DLT (0.45 ± 0.01) (t = 7.624, P < 0.01). The results of flow cytometry showed that the median fluorescence intensity of CD47 in macrophages after co-culture for 24 and 48 hours were 8 259.00 ± 66.01 and 9 445.00 ± 41.58, respectively, which were higher than the pre-co-culture value of 5 603.00 ± 193.40 (HSD = 0.691, 0.735; both P < 0.01). The median fluorescence intensity of SIRPα in macrophages after co-culture for 24 and 48 hours were 3 123.00 ± 184.60 and 2 931.00 ± 54.08, respectively, which were higher than the pre-co-culture value of 2 508.00 ± 43.15 (HSD = 0.491, 0.235; both P < 0.01). qPCR analysis revealed that the relative transcription levels of CD47 in macrophages after co-culture for 24 and 48 hours were 1.80 ± 0.02 and 1.64 ± 0.01, respectively, which were higher than the pre-co-culture level of 0.99 ± 0.01 (HSD = 0.098 and 0.125; both P < 0.01). The relative transcription levels of SIRPα in macrophages were 1.00 ± 0.02 before the addition of protoscoleces, 0.52 ± 0.05 and 1.27 ± 0.03 after co-culture for 24 and 48 hours, respectively. The level after 24 hours was lower than that before the addition of protoscoleces (HSD = 0.015, P < 0.01), while the level after 48 hours was higher than that before the addition of protoscoleces (HSD = 0.105, P < 0.01). Immunofluorescence analysis revealed that the Pearson correlation coefficients between CD47 and SIRPα after co-culture for 24 and 48 hours were 0.920 ± 0.001 and 0.990 ± 0.001, respectively, which were higher than the pre-co-culture value of 0.770 ± 0.001 (HSD = 0.091, 0.135; both P < 0.01). Fluorescence analysis revealed that the mean fluorescence intensity of EGFP in the inhibitor-infection group was 8 923.0 ± 49.3, which was higher than that in the infection group (7 537.0 ± 29.3) (HSD = 0.205, P < 0.01). Flow cytometry analysis revealed that the mean fluorescence intensity of EGFP in the inhibitor-infection group was 21.54 ± 0.03, which was higher than that in the infection group (18.55 ± 0.51) (HSD = 0.327, P < 0.01). qPCR analysis revealed that the mRNA relative transcription levels of inducible nitric oxide synthase (iNOS) and CD86, the markers of M1 macrophage, were 20.87 ± 0.40 and 40.64 ± 0.75 in the inhibitor-infection group, respectively, which were higher than 5.38 ± 0.11 and 3.79 ± 0.05 in the infection group (HSD = 0.194, 0.261; both P < 0.01). The mRNA relative transcription levels of arginase 1 (Arg1) and CD206, the markers of M2 macrophage, were 48.76 ± 2.22 and 6.33 ± 0.06 in the inhibitor-infection group, respectively, which were lower than 83.28 ± 0.58 and 12.33 ± 0.12 in the infection group (HSD = 0.283, 0.164; both P < 0.01). In the inhibitor-infection group, the mRNA relative transcription levels of interleukin 6 (IL-6), IL-1β and tumor necrosis factor α (TNF-α), the M1-type cytokines, were 3 896.00 ± 176.70, 271.30 ± 8.39 and 4.90 ± 0.10, respectively, which were higher than 2 869.00 ± 89.55, 154.90 ± 2.61 and 3.04 ± 0.03 in the infection group (HSD = 0.712, 0.625, 0.693; P < 0.05, 0.01, 0.01). The mRNA relative transcription levels of IL-10 and transforming growth factor β (TGF-β), the M2-type cytokines, in the inhibitor-infection group were 127.40 ± 4.92 and 1.34 ± 0.03, respectively, which were lower than 380.30 ± 8.55 and 1.61 ± 0.02 in the infection group (HSD = 0.324, 0.163; both P < 0.01). Conclusion The expression of CD47 and SIRPα in macrophages were increased after E. multilocularis infection, while the phagocytic function of macrophages was reduced. Specific inhibition of the CD47/SIRPα interaction could improve the phagocytic function and promote macrophage polarization towards the M1 phenotype.

Objective To investigate the notifiable parasitic diseases reported to the National Notifiable Infectious Disease Reporting System (NNIDRS) in Anhui Province from 2013 to 2023, and to analyze their epidemiological characteristics. Methods The demographic and clinical characteristics of cases with six notifiable parasitic diseases, including amoebic dysentery, schistosomiasis, malaria, visceral leishmaniasis, echinococcosis, and filariasis, and with current address in Anhui Province during the period from January 1, 2013, to December 31, 2023, were captured from the China Information System for Disease Control and Prevention. The temporal, spatial and population distributions of reported cases with six notifiable parasitic diseases, and the proportion of timely case reporting were analyzed with a descriptive epidemiological method, and the interval between disease diagnosis and case reporting to the NNIDRS was compared among groups with chi-square test, Fisher’s exact test, and Wilcoxon rank-sum test. Results A total of 25 699 cases with notifiable parasitic disease were reported in Anhui Province from 2013 to 2023, including 24 536 cases with schistosomiasis, 1 072 cases with malaria, 50 cases with amoebic dysentery, 6 cases with visceral leishmaniasis, 35 cases with echinococcosis, and no cases with filariasis were reported. The average annual incidence of notifiable parasitic diseases was 3.79/100 000 in Anhui Province from 2013 to 2023, and the annual incidence appeared an overall tendency towards a decline during the 11-year period. The incidence of notifiable parasitic diseases was higher among men (4.26/100 000) than among women (3.32/100 000) (χ² = 398.302, P < 0.05), and there were 60.96% (15 667/25 699) of cases with ages of 40 to 59 years. Farmer was the predominant occupation (92.70%, 23 824/25 699), and the incidence of notifiable parasitic diseases peaked during the period from June to November (76.26%, 19 598/25 699). The highest proportion of schistosomiasis cases was found in Anqing City (32.18%, 7 896/24 536), and the greatest proportion of malaria cases was recorded in Hefei City (52.15%, 559/1 072). The median interval between disease diagnosis and case reporting to the NNIDRS was 4.61 h, and the lowest percentage of timely case reporting was seen for echinococcosis (97.14%, 34/35). In addition, there were significant differences in the interval between disease diagnosis and case reporting (H = 26.559, P < 0.05) and the percentage of timely case reporting in terms of types of notifiable parasitic disease (χ² = 17.204, P < 0.05). Conclusion The prevalence of notifiable parasitic diseases was low in Anhui Province from 2013 to 2023, with middle-aged and elderly male farmers as high-prevalence populations. In addition, there was a region-specific incidence rate of notifiable parasitic diseases, and the proportion of echinococcosis cases reporting to the NNIDRS was low.

Objective To identify the species of Prosthogonimus in Grus japonensis and analyze the ribosomal DNA internal transcribed spacer (ITS) gene sequence. Methods Prosthogonimus samples were collected from the G. japonensis cloaca in Zhalong National Nature Reserve, Heilongjiang Province. The ITS gene of Prosthogonimus was amplified using PCR assay, and the amplified product was sequenced. The sequencing results were aligned with sequences recorded in the NCBI database using the BLAST tool. Repetitive sequences were identified using the Repeat Masker tool, and sequence similarity analysis was performed using the MegAlign software. Sequence alignments were conducted using the MEGA 7.0, Clustal X, and Paup software, and a phylogenetic tree was created using the maximum parsimony method. Results A total of 5 Prosthogonimus specimens were collected from G. japonensis, and five ITS gene sequences were obtained. Three sequences (GenBank accession numbers: PQ634967, PQ634968 and PQ634969) were 1 240 bp in size and showed 98.2% sequence similarity with P. cuneatus (GenBank accession number: OQ344776), which were identified as P. cuneatus. The other two sequences (GenBank accession numbers: PP956934 and PP956935) were 1 183 bp in size and showed 97.7% sequence similarity with P. pellucidus (GenBank accession number: KP192732), which were identified as P. pellucidus. There were no repetitive sequences in 5.8S or ITS2 genes of P. cuneatus and P. pellucidus, and there were 16 and 17 repetitive sequences the ITS1 gene of P. cuneatus and P. pellucidus, respectively. Phylogenetic analysis revealed that the P. cuneatus from this study was firstly clustered with P. cuneatus from Turdusmerula and Anasplatyrhynchos (Czech Republic isolate) into a subclade, and then clustered with P. pellucidus from this study into a clade. Conclusion The Prosthogonimus species infecting G. japonensis in the Zhalong National Nature Reserve are P. cuneatus and P. pellucidus, with 16 and 17 repetitive sequences in the P. cuneatus and P. pellucidus ITS1, respectively. P. cuneatus and P. pellucidus share a closer phylogenetic relationship with the Czech Republic isolate of P. cuneatus.

Objective To investigate the effect and mechanisms of Toxoplasma gondii type Ⅰ and Ⅲ rhoptry protein 16 (ROP16) on the proliferation and apoptosis of human monocytic leukemia THP-1 cells via TATA-binding protein-associated factor 15 (TAF15). Methods THP-1 cells were transfected with entiviruses overexpressing T. gondii type Ⅰ and Ⅲ ROP16 to generate cell lines that stably expressed ROP16 (THP-1-ROP16 Ⅰ/Ⅲ), and cell transfected with lentiviruses containing empty vectors (THP-1-Venus) served as an empty vector control, while non-transfected cells (THP-1) served as controls. The efficiency of overexpression was checked using quantitative real-time PCR (RT-qPCR) assay and Western blotting. The proteins interacting with ROP16 were identified using immunoprecipitation-mass spectrometry (IP-MS) in THP-1-ROP16 Ⅰ/Ⅲ cell lines, and the expression of the ROP16-interacting protein TAF15 was quantified using RT-qPCR and Western blotting assays. Three siRNA targeting different sites of TAF15 gene (siRNA 1215, siRNA 825, siRNA 288) were used to interfere with THP-1-ROP16 Ⅰ/Ⅲ cell lines and divided into THP-1-ROP16 Ⅰ/Ⅲ + siRNA 1215/825/288 groups, while an undisturbed control group (THP-1-ROP16 Ⅰ/Ⅲ + siRNA NC) was set up and the silencing efficiency of TAF15 was checked using Western blotting. In addition, the cell proliferation and apoptosis was measured using cell counting kit-8 (CCK-8) assay and flow cytometry, and the expression of cyclin-dependent kinase inhibitor 1A (p21), cyclin-dependent kinase 6 (CDK6), G1/S-specific cyclin (CyclinD1), B-cell lymphoma/leukemia-2 protein (Bcl-2), Bcl-2-associated X protein (Bax), cleaved caspase-3, caspase-9 and phosphorylated signal transducer and activator of transcription 3 (P-STAT3) was determined using Western blotting. Results The relative ROP16 mRNA expression was 2 679.427 ± 250.600 in the THP-1-ROP16 Ⅰ group and 2 395.410 ± 325.700 in the THP-1-ROP16 Ⅲ group, which was both higher than in the THP-1-Venus group (1.036 ± 0.102) (F = 153.3, P < 0.01), and the relative ROP16 protein expression was higher in the THP-1-ROP16 Ⅰ group (4.526 ± 0.020) and THP-1-ROP16 Ⅲ group (5.457 ± 0.250) than in the THP-1 Venus group (1.688 ± 0.653) (F = 76.4, P < 0.01). TAF15 was identified as a protein interacting with type Ⅰ and Ⅲ ROP16, and the relative TAF15 mRNA and protein expression was both higher in the THP-1 ROP16 Ⅰ group (6.027 ± 0.313 and 1.789 ± 0.145) and THP-1 ROP16 Ⅲ group (5.567 ± 0.088 and 1.593 ± 0.029) than in the THP-1 Venus group (0.985 ± 0.027 and 1.010 ± 0.365) (F = 869.4 and 50.6, P < 0.01). The relative TAF15 protein expression was 0.384 ± 0.047, 0.246 ± 0.072, and 0.125 ± 0.026 in the THP-1 ROP16 Ⅰ + siRNA 1215 group, the THP-1 ROP16 Ⅰ + siRNA 825 group, and the THP-1 ROP16 Ⅰ + siRNA 288 group 48 hours post-transfection with TAF15 siRNA, which was all lower than in the THP-1-Venus group (1.007 ± 0.019) (F = 313.1, P < 0.01), and the relative TAF15 protein expression was 0.186 ± 0.020, 0.180 ± 0.015, and 0.112 ± 0.019 in the THP-1 ROP16 Ⅲ + siRNA 1215 group, the THP-1 ROP16 Ⅲ + siRNA 825 group, and the THP-1 ROP16 Ⅲ + siRNA 288 group 48 hours post-transfection with TAF15 siRNA, which was all lower than in the THP-1 Venus group (0.995 ± 0.052) (F = 3 046.0, P < 0.01). CCK-8 assay measured the A₄₅₀ values of 0.803 ± 0.015 and 0.813 ± 0.011 in the THP-1-ROP16 Ⅰ + siRNA NC group and the THP-1-ROP16 Ⅲ + siRNA NC group, which were both lower than in the THP-1 Venus group (0.997 ± 0.010 and 0.995 ± 0.016) (t = 19.2 and 24.0, both P < 0.01). A₄₅₀ values of THP-1-ROP16 Ⅰ/Ⅲ + siRNA 825/288 groups were 0.986 ± 0.010, 0.983 ± 0.004; 0.980 ± 0.006, 0.984 ± 0.010 (F = 3.5, 2.9; both P > 0.05), respectively. The apoptotic rates of THP-1 cells were (38.19 ± 0.45)% in the THP-1-ROP16 Ⅰ + siRNA NC group and (38.06 ± 0.84)% in the THP-1-ROP16 Ⅲ + siRNA NC group, which were both higher than in the THP-1-Venus group [(28.41 ± 0.69)% ] (t = 20.5 and 17.7; both P < 0.01). The apoptotic rates of THP-1-ROP16 Ⅰ/Ⅲ + siRNA 825/288 group were (30.03 ± 1.83)%, (28.78 ± 0.72)%; (29.33 ± 0.80)%, (28.94 ± 0.58)% (F = 1.5, 0.4，both P > 0.05). The relative expression of p21, Bax, Caspase-9, Cleaved Caspase-3, and P-STAT3 proteins was 1.322 ± 0.027, 1.493 ± 0.030, 1.349 ± 0.021, 1.324 ± 0.020, and 10.500 ± 1.005 in the THP-1-ROP16 Ⅰ + siRNA NC group, which was all higher than in the THP-1-Venus group (1.000 ± 0.026, 0.996 ± 0.016, 0.989 ± 0.019, 0.994 ± 0.010 and 1.000 ± 0.001) (t = 14.8, 25.4, 22.3, 25.0 and 15.6; all P < 0.01), and the relative CDK6, CyclinD1, and Bcl-2 protein expression was 0.387 ± 0.040, 0.424 ± 0.030, and 0.438 ± 0.035 in the THP-1-ROP16 Ⅰ + siRNA NC group, which was all lower than in the THP-1-Venus group (0.989 ± 0.018, 1.000 ± 0.074 and 0.991 ± 0.016) (t = 23.6, 12.4 and 25.0; all P < 0.01). The relative expression of p21, Bax, caspase-9, cleaved caspase-3, and P-STAT3 proteins was 1.409 ± 0.020, 1.493 ± 0.030, 1.349 ± 0.021, 1.324 ± 0.020, and 16.210 ± 0.664 in the THP-1-ROP16 Ⅲ + siRNA NC group, which was all higher than in the THP-1-Venus group (1.004 ± 0.032, 0.996 ± 0.015, 0.989 ± 0.019, 0.994 ± 0.010 and 1.000 ± 0.001) (t = 18.7, 25.4, 22.3, 25.0 and 39.7; all P < 0.01), and the relative expression of CDK6, CyclinD1, and Bcl-2 proteins was 0.418 ± 0.021, 0.357 ± 0.040, and 0.411 ± 0.019 in the THP-1-ROP16 Ⅲ + siRNA NC group, which was all lower than in the THP-1-Venus group (1.000 ± 0.001, 1.001 ± 0.042 and 0.991 ± 0.016) (t = 47.7, 19.1 and 40.7; all P < 0.01). Conclusion T. gondii type Ⅰ and Ⅲ ROP16 proteins inhibit THP-1 cell proliferation and promote cell apoptosis through promoting TAF15 expression, which may be associated with inhibition of activation of the STAT3 signaling pathway in THP-1 cells.

Objective To identify differentially expressed genes (DEGs) in Dermatophagoides farinae post-treatment with horseradish using RNA sequencing, and to unravel the underlying mechanisms. Methods Ninety fumigation bottles were randomly divided into the experimental and control groups. In each bottle, 80 D. farinaes were placed back down on the adhesive plates of the fumigation bottles. D. farinaes were exposed to 25 μl of 0.25 ml/L horseradish dilutions in the experimental group, and paraffin in the control group for 24-hour fumigation. Then 2 000 survival D. farinae were collected from each group. All experiments were repeated 6 times, with triplicates for sequencing and others for real-time fluorescent quantitative PCR (qPCR) verification. RNA was extracted from D. farinae and reversely transcribed into cDNA, and a library was constructed for RNA sequencing. Following quality control and assembly, the sequencing data were aligned with non-redundant protein sequence (NR), gene ontology (GO), and Kyoto encyclopedia of genes and genomes (KEGG) databases, and the GO and KEGG functional annotations and enrichment analysis of DEGs were performed with the edge R software. In addition, DEGs identified in D. farinaes post-treatment with horseradish were sampled for qPCR assay, the qPCR data were subjected to analysis of variance with the R software. Results RNA sequencing yielded 56 625 transcripts with a total length of 72 418 610 nt, and a total of 39 510 UniGene clusters were yielded, with the highest number of clusters aligned to the NR database (25 450 clusters), followed by the evolutionary genealogy of genes: non-supervised orthologous groups (eggNOG） database (17 643 clusters), Swiss-prot database (13 338 clusters), GO database (13 234 clusters), and KEGG database (9 445 clusters), respectively. A total of 2 719 DEGs were identified in D. farinae post-treatment with horseradish, including 1 185 up-regulated genes and 1 534 down-regulated genes. GO annotations showed that the DEGs were mainly related to cellular processes, binding and catalytic activities, and GO enrichment analysis showed that the up-regulated genes were mainly enriched in ATP-binding cassette transporter G (ABCG) subfamily, cytochrome P450 (CYP450), heat shock proteins, and acetylcholine receptor and the down-regulated genes were mainly enriched in calmodulin and RNA polymerase. KEGG annotations showed that DEGs were mainly involved in the processes of transport and catabolism, signal transduction, and translation, and KEGG enrichment analysis revealed that the DEGs were significantly enriched in 30 pathways, which were mainly enriched in ribosome, microbial metabolism across diverse environments, and biosynthesis of secondary metabolites. qPCR results show that the relative transcription levels of UDP-glucuronosyltransferase (UGT) and multidrug resistance-associated protein 1 (MRP1) in the experimental group were 1.18 ± 0.22 and 6.22 ± 0.42, respectively, which were significantly upregulated compared to the control group (0.76 ± 0.30 and 2.52 ± 1.75) (F = 11.22, 16.83, both P < 0.05). And the relative transcription level of mitochondrial ribosome-associated GTPase (MTG) was 0.05 ± 0.04, which was significantly downregulated compared to the control group (1.23 ± 0.62) (F = 13.95, P < 0.05), which was consistent with RNA sequencing results. Conclusion There is differential expression of multiple genes in D. farinae post-treatment with horseradish, and UGT, CYP450, MRP1 and ABCG genes play important roles in the development of resistance to horseradish or detoxification metabolism of horseradish.

Echinococcosis is a parasitic disease that has been given a high priority for control interventions in China. Reference laboratories are a guarantee for accurate acquisition of epidemiological data, assessment of the effectiveness of control interventions and monitoring of disease prevalence. Since the establishment of the echinococcosis reference laboratory network was initiated in 2013 in China, the province-level reference laboratories have currently covered 70% of echinococcosis-endemic areas. This review summarizes the establishment of the national echinococcosis reference laboratories in China, analyzes the challenges and proposes future directions and priories, so as to provide insights into improving the capacity of echinococcosis detection and accelerating the process towards echinococcosis control in China.

Echinococcosis is a zoonotic parasitic disease caused by the larval stage of Echinococcus species. Echinococcus infection may modulate host immune response and alter host immune microenvironments, thereby triggering immune and inflammatory responses. As a pattern recognition receptor, Toll-like receptors (TLRs) may effectively recognize pathogen-associated molecular patterns and activate downstream signaling pathways by regulating TLR expression and functions, which play an important role in host immune responses to echinococcosis. This review summarizes the latest progress of TLRs in echinococcosis research, with a special emphasis on TLR2 and TLR4, and discusses their potential applications in future treatments of echinococcosis, so as to provide new insights into for echinococcosis prevention and treatment.

As a vector of multiple pathogens, sand flies have important epidemiological significance. A large number of microbial communities are colonized in the gut of sand flies, which pose a significant impact on the reproduction, growth and development of sand flies and sand flies-borne pathogens. The review summarizes the species of pathogens carried by sand flies and the composition of gut microbiota, and unravel the interactions among sand flies, pathogens, and gut microbiota. Gut microbiota not only directly inhibits the infection and transmission of pathogens, but also indirectly alters the ability of pathogen infections and transmission by sand flies through participation in the immune responses and defense mechanisms of sand flies.

Intestinal protozoa are single-celled eukaryotic microorganisms that are widely distributed in ecological environments. Most of the protozoan isolates are derived from excretions or secretions of individuals carrying cysts, which are used for experimental animal infections following purification and culture. Both the limitations of in vitro culture techniques and the host specificity of protozoa may directly affect the difficulty in the establishment of animal models. This review summarizes major in vitro culture techniques and appropriate animal models, and discusses the factors affecting the establishment of animal models. Considering that novel animal models are more effective to simulate the disease progression of human intestinal protozoan diseases, high attention should be paid to the development of experimental animal models of intestinal protozoan infections, notably novel animal models, so as to provide supports for the establishment and research of animal models of intestinal protozoan infections.

In order to investigate the trend in echinococcosis in Inner Mongolia from 2015 to 2023, all data pertaining to echinococcosis patients in Inner Mongolia from 2015 to 2023 were captured from the National Notifiable Disease Reporting System, and temporal, spatial and human distributions of echinococcosis were analyzed with a descriptive epidemiological method. A total of 500 echinococcosis cases were reported in 12 cities (leagues) of Inner Mongolia from 2015 to 2023. Echinococcosis cases were reported every year during the study period, with the highest number of cases reported in 2018 (104 cases, 21.0% of all cases), and the number of echinococcosis cases decreased significantly since 2021. In terms of regional distribution, Xilingol League (202 cases, 40.4% of all cases), Chifeng City (144 cases, 28.8%), and Hulun Buir City (52 cases, 10.4%) had the highest number of reported echinococcosis cases. There were 255 male echinococcosis cases and 245 female cases, with a male-to-female ratio of 1 ∶ 0.96, and the main occupations included herders (151 cases, 30.0%), farmers (145 cases, 29.0%), and housewives and unemployed (104 cases, 21.0%). The reported echinococcosis cases had ages of 5 to 97 years, and were concentrated at ages of 30 to 69 years, with the highest number of cases at 50 to 59 years (127 cases, 25.4%), followed by at ages of 40 to 49 years (116 cases, 23.2%). These data demonstrate that the number of reported echinococcosis cases peaked in Inner Mongolia in 2018 and decreased rapidly since 2021, indicating satisfactory effectiveness of the comprehensive echinococcosis control programme. Intensified management of the source of Echinococcus infections, surveillance on human echinococcosis and health education are recommended.

To investigate the origin of acquiring Leishmania infections and prevent the spread of visceral leishmaniasis, all cases with visceral leishmaniasis reported in Gaoping County, Shanxi Province from 2022 to 2023 which were recorded in the National Health Information System of China were subjected to case investigations, and host and vector surveys were performed in villages where visceral leishmaniasis cases lived. The sandfly density was investigated with light traps, and the serum anti-Leishmania antibody was detected in dogs with the rk39 immunochromatographic strips. In addition, foci disposal interventions were implemented in villages where the visceral leishmaniasis cases lived, including case finding, management of sources of infection, vector disposal and health education. During the period between 2022 and 2023, a total of 5 visceral leishmaniasis cases were reported in Gaoping County, Shanxi Province, and all were local cases, with main symptoms of fever, anemia and splenomegaly. The 5 cases were all adult males, and there were 3 workers and 2 farmers. The disease occurred throughout the year, and all cases lived in rural areas. A total of 608 sandflies were captured, with a mean density of 35.81 sandflies/(light•night), and a total of 244 domestic dogs were tested, with 24 dogs tested positive for anti-Leishmania antibody (9.8%). The vector Phlebotomus chinensis and dogs infected with Leishmania were detected in all villages where 5 visceral leishmaniasis cases lived. A total of 25 blood samples were collected from 5 cases’ neighbors, and all were tested negative for anti-Leishmania antibody. Foci disposal interventions included sacrificing of all 24 Leishmania-infected dogs, residual spray with deltamethrin insecticides for interior and exterior walls of visceral leishmaniasis cases’ houses and all sheep pens, dog houses and shabby cave dwellings in villages where cases lived (covering approximately 3 000 m²), 10 health education lectures pertaining to visceral leishmaniasis control knowledge and more than 5 000 poster foldouts pertaining to visceral leishmaniasis control knowledge in villages where cases lived. These findings indicate local transmission of visceral leishmaniasis in Gaoping County, with a high sandfly density and a high prevalence rate of Leishmania infections in dogs. Dog management and vector control are recommended to prevent the further spread of visceral leishmaniasis.

A statistical analysis was performed on the scrub typhus surveillance data in Baoshan City, Yunnan Province from January 1, 2017 to December 31, 2023 reported to the Chinese Disease Prevention and Control Information System. The concentration ratio (M value) of scrub typhus was calculated to analyze the seasonal distribution characteristics, and the day of the peak incidence of scrub typhus and epidemic peaks of scrub typhus were estimated using the circular distribution method. A Pearson correlation analysis was performed on the M value and the concentration parameter (r value) of a circular distribution. A total of 11 537 cases with scrub typhus were reported in Baoshan City from 2017 to 2023, with an annual reported incidence rate of 65.12/105, and the highest incidence in 2023 (96.71/105), and the incidence appeared an overall tendency towards a rise from 2017 to 2023 (χ² = 484.882, P < 0.01). Farmers were the primary occupation among all cases with scrub typhus (78.79%, 9 090/11 537), and the reported scrub typhus cases are concentrated during the period between July and October from 2017 to 2023, with the highest proportion of reported cases in August each year, and the numbers of reported scrub typhus cases accounted for 26.74%, 33.13%, 34.25%, 32.84%, 34.55%, 27.17% and 39.34% of total cases each year from 2017 to 2023, which accounted for 33.04% (3 812/11 537) of total cases. The radar chart appeared a unimodal distribution in each year and each month, which met the prerequisite for concentration ratio and circular distribution analyses. The M value of total scrub typhus cases was 0.779 from 2017 to 2023, with 0.729 to 0.814 each year during the study period, indicating a strong seasonality of scrub typhus incidence in Baoshan City. The r value of the circular distribution of scrub typhus incidence was 0.758, with $\bar{\alpha } $ of 227.38° (184.89°, 269.87°) (Z = 6 626.66, P < 0.001) indicating the day of peak incidence on August 21, and epidemic peak during the period from July 6 to September 30. In addition, Pearson correlation analysis revealed a strong positive correlation between the M value and r value (Pearson coefficient was 0.940 and P was 0.001 65).

The Affiliated Hospital of Qinghai University admitted a 53-year-old male patient in August 2023, who reported that he usually had no special discomfort, no symptoms such as radiating pain in the shoulder and back and abdominal pain, and no history of close contact with animals such as cattle, sheep, and dogs or living in a long-term endemic area. Physical examination and laboratory tests after admission were not abnormal. Combined with the patient’s medical history and imaging examinations such as three-phase dynamic-enhanced CT of the abdominal organs, the preliminary diagnosis was pancreatic plasma cystadenoma. After excluding relevant contraindications to surgery, laparoscopic pancreatic body-tail resection was performed. Intraoperatively, it was found that there was no ascites in the abdominal cavity, and the mass was located in the body of the pancreas, with unclear borders, protruding from the surface of the pancreas, measuring about 4.0 cm × 3.7 cm × 3.0 cm, with poor mobility and adhesion to the surrounding intestinal tract. The liver was normal in texture and color. After complete resection of the caudal part of the pancreatic body and the lesion, intraoperative frozen pathology suggested cystic echinococcosis of the pancreatic body. The patient’s postoperative vital signs were stable and recovery was uneventful. One month later, a follow-up CT scan of the upper abdomen showed that the operated area was better than before, and the patient reported that he had recovered well after the operation, with no obvious special discomfort, normal diet and sleep, normal urination and defecation, and the rest of the patient did not see any obvious abnormality.