Internet addiction and depressive and anxious symptoms among Chinese rural left-behind adolescents: Mediating roles of resilience and friendship quality.

BACKGROUND: Rural left-behind adolescents are more vulnerable to Internet addiction and depressive and anxious symptoms due to the lack of family support and parental supervision. This study was the first to investigate the longitudinal relationships between Internet addiction and depressive and anxious symptoms and to examine the mediating roles of resilience and friendship quality in rural left-behind adolescents. METHODS: Included in this study, which was from a longitudinal study conducted five times over 2 years, were 1001 rural left-behind adolescents. The internationally used scales for depressive and anxious symptoms, Internet addiction, resilience and friendship quality were administered. A structural equation model was used for analysis. RESULTS: The prevalence of Internet addiction, depressive and anxious symptoms among rural left-behind adolescents were 17.7%, 35.8% and 27.6%, respectively. Internet addiction predicted the later depressive and anxious symptoms (ß = 0.200, 95% confidence interval [CI]: 0.116-0.274 and ß = 0.263, 95% CI: 0.188-0.330). Resilience acted as an independent mediator in the relationships between Internet addiction and depressive and anxious symptoms (ß = 0.037 and 0.034, P < 0.01). Resilience and friendship quality played a chain-mediating role on the longitudinal relationships between Internet addiction and depressive and anxious symptoms (ß = 0.011 and 0.010, P < 0.001). The mediating effects accounted for 24.0% and 16.7% of the total effects, respectively. CONCLUSION: Resilience and friendship quality play an independent or chain-mediating role in longitudinal relationships between Internet addiction and depressive and anxious symptoms. The findings inform targeted intervention strategies to improve the mental health of left-behind adolescents.

[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.

A Macrophage Differentiation-Mediated Gene: DDX20 as a Molecular Biomarker Encompassing the Tumor Microenvironment, Disease Staging, and Prognoses in Hepatocellular Carcinoma.

Background: DDX20 involves the mechanism of cell proliferate, mitogenic Ets transcriptional suppressor (METS), which can arrest the cell cycle of macrophages. However, little is known about DDX20 expression, clinical values, and the relationship with tumor microenvironment in HCC. Methods: We mined the transcriptional, protein expression and survival data of DDX20 in HCC from online databases. The immunological effects of DDX20 were estimated by bioinformatic algorithms. The RNAi and CRISPR screening were used to assess the gene effect of DDX20 for the EGFR gene in liver tumor cell. Results: We found that the DDX20 was highly expressed in HCC. The qRT-PCR result shows a significantly upregulated DDX20 expression in HCC samples from the West China Hospital. The high mRNA expression of DDX20 is associated with a poor survival. DDX20 expression is positively correlated with MDSCs in HCC tissues. Moreover, DDX20 has a high predicted ability for the response to immunotherapy. Furthermore, hsa-mir-324-5p could regulate the macrophage differentiation by interacting with DDX20. Meanwhile, the EGFR gene gets a high dependency score for DDX20. Conclusion: In sum, DDX20 may serve as a prognostic marker for worse clinical outcomes with HCC and potentially enable more precise and personalized immunotherapeutic strategies in the future.

Assessment of echinococcosis control in Tibet Autonomous Region, China.

BACKGROUND: In China the highest prevalence of echinococcosis is in Tibet Autonomous Region (TAR). The government has issued documents and implemented comprehensive prevention and control measures focusing on controlling the source of infection of echinococcosis. It was very important to understand the implementation and effect of infectious source control measures. The purpose of this study was to examine the implementation of measures to control infectious source (domestic and stray dogs) in TAR and to assess their effectiveness. METHODS: We collected data on domestic dog registration and deworming and stray dog sheltering in 74 counties/districts in the TAR from 2017 to 2019. Fecal samples from domestic dogs were collected from randomly selected towns to determine Echinococcus infection in dogs using coproantigen ELISA. We analyzed the data to compare the canine rate of infection between 2016 and 2019. The data analysis was performed by SPSS statistical to compare dog infection rate in 2016 and 2019 by chi-square test, and ArcGIS was used for mapping. RESULTS: From 2017 to 2019, 84 stray dog shelters were built in TAR, and accumulatively 446,660 stray or infected dogs were arrested, sheltered, or disposed of. The number of domestic dogs went downward, with an increased registration management rate of 78.4% (2017), 88.8% (2018), and 99.0% (2019). Dogs were dewormed 5 times in 2017, 12 times in 2018, and 12 times in 2019. The dog infection rate was 1.7% (252/14,584) in 2019, significantly lower than 7.3% (552/7564) from the survey of echinococcosis prevalence in Tibet in 2016 (P < 0.05). CONCLUSION: Between 2017 and 2019, the number of stray dogs and infection rate of Echinococcus spp. in domestic dogs decreased significantly, indicating that dogs were effectively controlled as a source of infection in TAR and reflecting a significant decrease in the risk of echinococcosis transmission.

Echinococcus spp. and genotypes infecting humans in Tibet Autonomous Region of China: a molecular investigation with near-complete/complete mitochondrial sequences.

BACKGROUND: Molecular markers are essential to identify Echinococcus species and genotypes in areas with multiple Echinococcus species to understand their epidemiology and pathology. Tibet Autonomous Region (TAR) is one of the areas worst hit by echinococcosis. However, molecular epidemiology is still missing among echinococcosis patients in TAR. This research explored the Echinococcus species and genotypes infecting humans in TAR and the population diversity and the possible origin of G1 in TAR. METHODS: Cyst samples were collected in one echinococcosis-designated hospital in TAR. Echinococcus species and genotypes were identified through a maximum-likelihood approach with near-complete/complete mtDNA using IQ-TREE. Phylogenetic networks were built with PopART, and the phylogeographical diffusion pattern was identified using a Bayesian discrete phylogeographic method. RESULTS: Using phylogenetic trees made with near-complete/complete mtDNA obtained from 92 cysts from TAR patients, the Echinococcus species and genotypes infecting humans in TAR were identified as Echinococcus granulosus (s.s.) G1 (81, 88.04%), accounting for the majority, followed by G6 of the E. canadensis cluster (6, 6.52%), E. granulosus (s.s.) G3 (3, 3.26%), and E. multilocularis (2, 2.17%). An expansion trend and a possible recent bottleneck event were confirmed among the G1 samples in TAR. Adding the other near-complete mtDNA of G1 samples globally from the literature, we identified the possible phylogeographic origin of the G1 samples in TAR as Turkey. CONCLUSIONS: Using near-complete/complete mtDNA sequences of Echinococcus spp. obtained from echinococcosis patients, a variety of Echinococcus species and genotypes infecting humans throughout TAR were identified. As far as we know, this is the first comprehensive molecular investigation of Echinococcus species and genotypes infecting humans throughout TAR. We identified, for the first time to our knowledge, the possible origin of the G1 in TAR. We also enriched the long mtDNA database of Echinococcus spp. and added two complete E. multilocularis mtDNA sequences from human patients. These findings will improve our knowledge of echinococcosis, help to refine the targeted echinococcosis control measures, and serve as a valuable baseline for monitoring the Echinococcus species and genotypes mutations and trends of the Echinococcus spp. population in TAR.

Geographic distribution and prevalence of human echinococcosis at the township level in the Tibet Autonomous Region.

BACKGROUND: Echinococcosis, a zoonotic parasitic disease, is caused by larval stages of cestodes in the Echinococcus genus. Echinococcosis is highly prevalent in ten provinces/autonomous regions of western and northern China. In 2016, an epidemiological survey of Tibet Autonomous Region (TAR) revealed that the prevalence of human echinococcosis was 1.66%, which was much higher than the average prevalence in China (0.24%). Therefore, to improve on the current prevention and control measures, it is important to understand the prevalence and spatial distribution characteristics of human echinococcosis at the township level in TAR. METHODS: Data for echinococcosis cases in 2018 were obtained from the annual report system of echinococcosis of Tibet Center for Disease Control and Prevention. Diagnosis had been performed via B-ultrasonography. The epidemic status of echinococcosis in all townships in TAR was classified according to the relevant standards of population prevalence indices as defined in the national technical plan for echinococcosis control. Spatial scan statistics were performed to establish the geographical townships that were most at risk of echinococcosis. RESULTS: In 2018, a total of 16,009 echinococcosis cases, whose prevalence was 0.53%, were recorded in 74 endemic counties in TAR. Based on the order of the epidemic degree, all the 692 townships were classified from high to low degrees. Among them, 127 townships had prevalence rates ≥ 1%. The high prevalence of human echinococcosis in TAR, which is associated with a wide geographic distribution, is a medical concern. Approximately 94.65% of the villages and towns reported echinococcosis cases. According to spatial distribution analysis, the prevalence of human echinococcosis was found to be clustered, with the specific clustering areas being identified. The cystic echinococcosis primary cluster covered 88 townships, while that of alveolar echinococcosis's covered 38 townships. CONCLUSIONS: This study shows spatial distributions of echinococcosis with different epidemic degrees in 692 townships of TAR and high-risk cluster areas at the township level. Our findings indicate that strengthening the echinococcosis prevention and control strategies in TAR should directed at townships with a high prevalence and high-risk clustering areas.

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.

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.

Characterizing dynamic changes of plasma cell-free Echinococcus granulosus DNA before and after cystic echinococcosis treatment initiation.

Over one million people are living with cystic echinococcosis (CE) and alveolar echinococcosis (AE). For CE, long-term albendazole treatment is often needed, which requires regular follow-up. Follow-up is mainly through imaging which is insensitive to subtle changes and subjective to experience. We investigated the changes of Echinococcus granulosus (Eg) cell-free DNA (cfDNA) in plasma of CE patients before and after albendazole treatment to evaluate its potential as an objective marker for treatment follow-up. Plasma samples of nine CE patients were collected before and after treatment. We identified Eg cfDNA from every sample through high-throughput sequencing. Eg cfDNA concentration and fragment length increased significantly after the treatment period. Ultrasound examination before and after the treatment initiation reflected the drug effects to a certain extent, as the cyst size of four patients reduced. Our findings indicated that Eg cfDNA from plasma could be a potential marker in the monitoring of CE treatment.

Reducing Canine Echinococcus Infection with Smart Deworming Collars - Tibet, China, June-November, 2020.

What is already known about this topic? Existing manual deworming programs launched have made great progress in reducing the Echinococcus infection rate of domestic dogs, but significant challenges remain in scattered nomadic communities inhabiting the Tibetan Plateau. The low deworming frequency and low levels of coverage were responsible for the high infection rate of Echinococcus spp. among dogs. What is added by this report? Smart deworming collars controlled by a remote management system (RMS) was found to increase the deworming frequency and coverage and subsequently reduce the canine infection rates with Echinococcusspp.. What are the implications for public health practice? As an innovative tool, smart deworming collars may drive the paradigm shift from manual deworming to smart deworming and stop the transmission of echinococcosis.

[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.

A rapid high-resolution melting method for differentiation of Leishmania species targeting lack gene.

OBJECTIVES: The aim of this research is to verify that if lack gene can be used for differentiation of Leishmania under HRM assay. METHODS: Two specific primers were designed targeting polymorphic sites on the lack gene sequence. DNA from promastigotes of six species of Leishmania based on reference strains were tested following a HRM protocol. We also tested ten Chinese isolates in blind to validate our method. RESULTS: Combined with amplicon of the two primers, the six reference strains can be easily discriminated without the effect of initial concentration of DNA templates. Ten Chinese isolates detected by our HRM method resulted in full accord with the standard identification results in previous study. CONCLUSION: HRM is a rapid and reproducible method to discriminate different Leishmania species and lack gene is a potential novel biological characteristic for easy differentiation of Leishmania isolates in China.