[Relationship Between the Level of Anatomical Risk of Hepatic Alveolar Echinococcosis and Complications after Radical Resection].

Objective: To analyze the impact of high or low levels of anatomical risk of hepatic alveolar echinococcosis (HAE) on complications after radical resection. Methods: The baseline, surgical, and complication data were retrospectively collected from hepatic alveolar echinococcosis patients who underwent radical resection at the Ganzi Branch Hospital, West China Hospital, Sichuan University from 2015 to 2022. The patients were divided into anatomical low-risk (ALR) and anatomical high-risk (AHR) groups based on the PNM staging system designed by the World Health Organization (WHO-PNM). Complications were analyzed according to the Clavien-Dindo classification. Univariate and multivariate logistic regression analyses were performed to assess the effect of high and low risks of lesion anatomy on complications. Results: Radical surgery was performed in 216 HAE patients and 102 of whom were in the AHR group. The median operative time was 230 (175, 300) min, the median intraoperative blood loss was 600 mL, and 129 (59.7%) patients developed complications. The complication rate was 73.5% (75/102) in the AHR group and 47.4% (54/114) in the ALR group, demonstrating statistically significant difference ( P<0.05). The incidence of serious complications was 36.3% (37/102) in the AHR group and 13.2% (15/114) in the ALR group, demonstrating statistically significant difference ( P<0.05). There was significant difference in the proportions of patients having postoperative complications of bile leak, anemia, fever, pleural effusion and ascites between the AHR group and the ALR group ( P<0.05). Multivariate logistic regression analysis suggested that AHR was the only independent risk factor for complications, including bile leak, anemia, fever, and pleural effusion, and severe complications. Conclusion: The anatomical risk of hepatic alveolar echinococcosis is independently associated with the development of multiple postoperative complications, and physicians should choose surgical procedures cautiously according to the actual situation when dealing with patients defined as AHR according to WHO-PNM.

[Preliminary Study on PCR Method for Laboratory Diagnosis with Fecal Specimens of Entamoeba histolytica Infection].

OBJECTIVE: To develop a PCR method for Entamoeba histolytica（ E.histolytica) detection in fecal specimens, and to compare the performance of PCR to that of microscopy and ELISA. METHODS: Two pairs of self-designed primers and 2 pairs of primers from references based on small subunit ribosome RNA (SSU rRNA) fragment of E. histolytica standard strain were synthetized. DNA from E. histolytica reference strain were amilified by the conventional PCR using the 4 pairs of primers. 221 stool samples from diarrhea patients were collected and detected for E. histolytica by three methods: Entamoeba trophozoites and cysts detection by microscopy, E. histolytica-specific antigen detection using enzyme-linked immunosorbent assay (ELISA) kit ( E. HISTOLYTICA II), amplification of SSU rRNA fragment of E. histolytica by PCR method. Positive rate of three methods were compared by chi-square test, and Kappa test was applied to determine the concordance among the three methods. RESULTS: Specific fragments of E. histolytica were amplified by the PCR method we developed in this study. Positive rates of PCR, microscopy and ELISA were 2.26%, 0.90% and 9.50%, respectively. The positive rates of the three methods were significantly different ( χ 2 =23.34, P<0.01). The Kappa value of PCR and microscopy was 0.216, and that of PCR and ELISA method was -0.134, both of which showed a weak consistency. PCR results showed best consistency with clinical diagnosis. CONCLUSION: The PCR method we established in this study has a better performance in accuracy than microscopy and ELISA have in laboratory diagnosis of E. histolytica infection.

Effects of management of infection source of echinococcosis in Linzhi, Tibet Autonomous Region of China.

BACKGROUND: Echinococcosis is highly endemic in western and northern China. Tibet Autonomous Region (TAR) is the most serious prevalent area. Linzhi is located in southeastern part of TAR. Dogs are the primary infection source for the transmission of echinococcosis to humans. A control and prevention campaign based on dog management has been implemented in the past three years. This study aims to evaluate the effects of dog management on the infection rate of dogs. METHODS: Data of dog population, registration and de-worming of seven counties/district in Linzhi between 2017 and 2019 were obtained from the annual prevention and control report. Domestic dog fecal samples were collected from each endemic town of seven counties/district in Linzhi in 2019 to determine the infection of domestic dogs using coproantigen enzyme-linked immunosorbent assay (ELISA). Data analysis was processed using SPSS statistics to compare dog infection rate between 2016 and 2019 by chi-square test, and maps were mapped using ArcGIS. RESULTS: In Linzhi, domestic dog population has decreased from 17 407 in 2017 to 12 663 in 2019, while the registration rate has increased from 75.9% in 2017 to 98.6% in 2019. Similarly, stray dog population has decreased from 14 336 in 2017 to 11 837 in 2019, while sheltered rate has increased from 84.6% in 2017 to 96.6% in 2019. Dog de-worming frequency has increased from 4 times per annum in 2017 to 12 times in 2019, indicating that approximately every dog was dewormed monthly. A total of 2715 dog fecal samples were collected for coproantigen ELISA assay. The dog infection rate was 2.8% (77/2715) in 2019, which was significantly lower than 7.3% (45/618) in 2016 (P < 0.05). CONCLUSIONS: Increased dog registration, decreased dog population, and increased dog de-worming frequency contributed to significantly decrease the dog infection rate in Linzhi. Control and prevention campaign based on dog management could significantly decrease dog infection with Echinococcus spp. in echinococcosis endemic areas.

Anti-echinococcal effect of verapamil involving the regulation of the calcium/calmodulin-dependent protein kinase II response in vitro and in a murine infection model.

BACKGROUND: Echinococcosis, which is caused by the larvae of cestodes of the genus Echinococcus, is a parasitic zoonosis that poses a serious threat to the health of humans and animals globally. Albendazole is the drug of choice for the treatment of echinococcosis, but it is difficult to meet clinical goals with this chemotherapy due to its low cure rate and associated side effects after its long-term use. Hence, novel anti-parasitic targets and effective treatment alternatives are urgently needed. A previous study showed that verapamil (Vepm) can suppress the growth of Echinococcus granulosus larvae; however, the mechanism of this effect remains unclear. The aim of the present study was to gain insight into the anti-echinococcal effect of Vepm on Echinococcus with a particular focus on the regulatory effect of Vepm on calcium/calmodulin-dependent protein kinase II (Ca2+/CaM-CaMKII) in infected mice. METHODS: The anti-echinococcal effects of Vepm on Echinococcus granulosus protoscoleces (PSC) in vitro and Echinococcus multilocularis metacestodes in infected mice were assessed. The morphological alterations in Echinococcus spp. induced by Vepm were observed by scanning electron microscopy (SEM), and the changes in calcium content in both the parasite and mouse serum and liver were measured by SEM-energy dispersive spectrometry, inductively coupled plasma mass spectrometry and alizarin red staining. Additionally, the changes in the protein and mRNA levels of CaM and CaMKII in infected mice, and in the mRNA levels of CaMKII in E. granulosus PSC, were evaluated after treatment with Vepm by immunohistochemistry and/or real-time quantitative polymerase chain reaction. RESULTS: In vitro, E. granulosus PSC could be killed by Vepm at a concentration of 0.5 µg/ml or higher within 8 days. Under these conditions, the ultrastructure of PSC was damaged, and this damage was accompanied by obvious calcium loss and downregulation of CaMKII mRNA expression. In vivo, the weight and the calcium content of E. multilocularis metacestodes from mice were reduced after treatment with 40 mg/kg Vepm, and an elevation of the calcium content in the sera and livers of infected mice was observed. In addition, downregulation of CaM and CaMKII protein and mRNA expression in the livers of mice infected with E. multilocularis metacestodes was found after treatment with Vepm. CONCLUSIONS: Vepm exerted a parasiticidal effect against Echinococcus both in vitro and in vivo through downregulating the expression of Ca2+/CaM-CaMKII, which was over-activated by parasitic infection. The results suggest that Ca2+/CaM-CaMKII may be a novel drug target, and that Vepm is a potential anti-echinococcal drug for the future control of echinococcosis.

Smart deworming collar: A novel tool for reducing Echinococcus infection in dogs.

Echinococcosis is a serious zoonotic parasitic disease transmitted from canines to humans and livestock. Periodic deworming is recommended by the WHO/OIE as a highly effective measure against echinococcosis. However, manual deworming involves significant challenges, particularly in remote areas with scarce resources. The insufficient awareness delivering praziquantel (PZQ) baits for dogs leads to low compliance rate. The aim of this study was therefore to develop a novel smart collar for dogs to address these challenges. We developed a smart Internet of Things (IoT)-based deworming collar which can deliver PZQ baits for dogs automatically, regularly, quantitatively with predominant characteristics of being waterproof, anti-collision, cold-proof and long life battery. Its performance was tested in two remote locations on the Tibetan Plateau. A cross-sectional survey was conducted to evaluate the compliance of the dog owners. Further, a randomized controlled study was performed to evaluate the difference between smart-collar deworming and manual deworming. The collar's effectiveness was further assessed on the basis of Generalized Estimation Equations (GEE). The testing and evaluation was done for 10 smart deworming collars in factory laboratory, 18 collars attached for 18 dogs in Seni district, Tibet Autonomous Region, China, and 523 collars attached for 523 dogs in Hezuo city, Gansu province, China. The anti-collision, waterproof, and coldproof proportion of the smart collars were 100.0%, 99.5%, and 100.0%, respectively. When compared to manual deworming, the dogs' risk of infection with Echinococcus on smart-collar deworming is down to 0.182 times (95% CI: 0.049, 0.684) in Seni district and 0.355 (95%CI: 0.178, 0.706) in Hezuo city, the smart collar has a significant protective effect. The owners' overall compliance rate to attach the smart collars for their dogs was 89%. The smart deworming collar could effectively reduce the dogs' risk of infection with Echinococcus in dogs, significantly increase the deworming frequency and coverage and rapidly remove worm biomass in dogs. Thus, it may be a promising alternative to manual deworming, particularly in remote areas on the Tibetan Plateau.