Clonorchis Research Articles

PREVALENCE AND INTENSITY OF INTESTINAL PARASITIC WORM EGGS IN CATTLE FECES AT THE SLAUGHTERHOUSE IN TANAH MERAH VILLAGE, NORTH SAMARINDA DISTRICT

Cattle farming is the most popular business for many people as a source of income because it producer meat and milk that has high economic value. Based on the maintenance aspect, cattle products produced in Indonesia have not been maximized due to many health problems of farm animals. One of the pathogens that are often found on farms is parasites. This can pose a real danger to some livestock. This study aims to determine the prevalence and intensity of intestinal parasitic worm eggs in cattle feces found in Tanah Merah Village Slaughterhouse, North Samarinda District. The total number of samples in the study was 30 cattle fecal samples. Worm eggs are observed through fecal examination using two methods, namely sedimentation method and flotation method. The results of this study showed that 13 types of intestinal parasitic worm eggs were found in cattle feces found from Tanah Merah Village Slaughterhouse, Samarinda, namely Ascaris lumbricoides, Bunostomum phlebotomum, Clonorchis sinensis, Dicrocelium dendriticum, Fasciola hepatica, Hymenelopsis nana, Paragonimus wastermani, Schistosoma bovis, Schistosoma mansoni, Schistosoma haemabotium, Strongyloides stecoralis, Taenia saginata and Trichuris trichura. Taenia saginata worm eggs have the highest intensity value of 1.028 eggs/individual. Ascaris lumbricoides worm eggs have the highest prevalence value of 26,67% of the total individuals.

Clonorchis sinensis extracellular vesicles associated with Csi-let-7a-5p activate pro-inflammatory macrophages to induce biliary injury.

During Clonorchis sinensis (C. sinensis) infection, pro-inflammatory macrophages (M1 macrophages) are highly activated, yet their role in the disease remains poorly understood. Previous studies have demonstrated that extracellular vesicles from C. sinensis (CsEVs) can activate these macrophages, and inhibiting a specific miRNA (Csi-let-7a-5p) in CsEVs (InCsEVs) can reduce this activation. In the present study, liver macrophages in mice were removed using clodronate liposomes (Clodlip). Subsequently, different types of bone marrow-derived macrophages (BMDMs) were adoptively transferred into the mice lacking liver macrophages: untreated (PBS-BMDM), treated with CsEVs (CsEVs-BMDM), treated with a control (ScrCsEVs-BMDM), or treated with InCsEVs (InCsEVs-BMDM). Biliary damages were then evaluated. The results indicated that the transferred macrophages successfully repopulated the mice. CsEVs-BMDM led to significant inflammation and bile duct damage, accompanied by higher levels of inflammatory cytokines (TNF-α and IL-1β). However, when macrophages were treated with InCsEVs, the damage and inflammation were alleviated, and the levels of TNF-α and IL-1β decreased. These findings suggest that pro-inflammatory macrophages activated by CsEVs, especially through Csi-let-7a-5p, play a crucial role in biliary damage during C. sinensis infection. Although other immune cells may also be involved, this study emphasizes the significance of pro-inflammatory macrophages in clonorchiasis.

Formation of non-cholesterol gallstones in populations within clonorchiasis endemic regions is closely related to Clonorchis sinensis infection

BackgroundNumerous risk factors are linked to gallbladder stone disease (GBSD). Nonetheless, the relationship between Clonorchis sinensis (C. sinensis) and this condition remains to be clarified.MethodsThe antibody against C. sinensis in serum and the glucose, triglyceride, and cholesterol levels were investigated in 220 patients with GBSD and 251 controls. Bile components were analysed in patients with gallbladder polyps (GP, n = 18), gallstones (GS, n = 265), and GS combined with C. sinensis infection (GSI, n = 243). Additionally, the gallbladder ejection fraction (%E), residual gallbladder volume (RGV) at 1 h after a fatty meal, and fasting gallbladder volume (FGV) were compared among the GP (n = 43), GS (n = 311), and GSI (n = 277) groups.ResultsThe results indicated positive antibody against C. sinensis (OR: 1.759, 95% CI: 1.163–2.662) and hyperglycaemia (glucose concentration ≥ 6.10 mmol/L, OR: 2.263, 95% CI: 1.227–4.172) were risk factors for GBSD. There were more non-cholesterol stones in GSI patients (216/241, 89.6%) than in GS patients (137/281, 48.8%) (P < 0.0001). Microscopic observations revealed that mucus containing glycogen coated the C. sinensis eggs and the proportion of dead eggs gradually increased in bile, sediment, and stones alongside rising calcium salt content. Total bile acid and cholesterol concentrations were lower in GSI patients than in GP patients or GS patients (P < 0.05). Moreover, increased FGV and RGV and decreased %E were observed in GSI patients compared with GP patients (P < 0.001).ConclusionsThe formation of non-cholesterol gallstones in populations residing in endemic areas is related to the deposition, death, and calcification of eggs in the gallbladder, changes in bile components, and decreased gallbladder motility caused by C. sinensis infection.

Research of the clinical features, risk factors, and surgical diagnosis of intramural stones in patients with gallbladder stones

Crystals or stones within the gallbladder wall in patients with gallbladder stones (GBS) have been occasionally reported, but their clinical features and aetiology remain unclear. This retrospective study analysed 323 consecutive patients with GBS who underwent rigid choledochoscopic gallbladder-preserving cholecystolithotomy to determine the detection rate, clinical features, and potential risk factors of gallbladder intramural stones (IS). IS were found in 24.1% (78/323) of patients, characterised by distinct cholangioscopic findings, including stone shadows, yellow floating bands, or a combination of both within the gallbladder wall. Compared to patients without IS, those with IS had a higher prevalence of Clonorchis sinensis (C. sinensis) eggs (60.3% vs. 40.8%, P < 0.05) and elevated serum cholesterol, LDL cholesterol, and Apo-B levels (P < 0.05). However, stone composition and C. sinensis egg detection rates did not differ between intraluminal stones and IS within the same patient (P > 0.05). Logistic regression analysis revealed that IS were associated with C. sinensis infection and elevated Apo-B levels. In conclusion, IS share homology with intraluminal stones in the same patient with GBS and exhibit unique appearances in rigid choledochoscopy. For patients with GBS and IS, elevated serum Apo-B levels and C. sinensis infection were independent risk factors.

Proteomic profiling of spleen in rat infected with clonorchis sinensis using liquid chromatography tandem mass spectrometry analysis.

Proteomic profiling of spleen in rat infected with clonorchis sinensis using liquid chromatography tandem mass spectrometry analysis.

Development and evaluation of fluorescent recombinase polymerase amplification (RPA)-based method for rapid detection of Necator americanus.

Necator americanus is the predominant species causing hookworm infections in humans. Despite advancements in prevention strategies, mild cases of infection still occur, highlighting the need for improved detection technology. Recombinase Polymerase Amplification (RPA) is an isothermal molecular diagnostic known for its sensitivity, speed, portability, and widespread application in detecting various pathogens. Although several molecular assays are available for N. americanus, they have limitations in detecting mild N. americanus infections. Fluorescent RPA primers and probes targeting the N. americanus internal transcribed spacer 2 (ITS2) gene were developed. The method's detection limit was assessed via serial dilution of genomic DNA. Specificity was confirmed against Clonorchis sinensis, Schistosoma japonicum, Fasciola hepatica, Ascaris lumbricoides, Enterobius vermicularis and Ancylostoma duodenale. Thirty samples identified as positive by Kato-Katz, along with 11 samples identified as negative by the method, were tested to evaluate the sensitivity and specificity of fluorescent RPA. Additionally, 287 field samples were tested for validation with these methods. All positive samples were identified as either N. americanus or A. duodenale. This study successfully developed a fluorescent RPA assay targeting the ITS2 gene of N. americanus. The length of the amplified fragment was 237 bp. Optimized conditions were achieved, resulting in a minimum detection limit of 1fg/µL, with no cross-reactivity with other pathogens. In laboratory validation, the fluorescent RPA assay demonstrated 100% sensitivity (30/30) and 100% specificity (11/11) compared to the Kato-Katz, and 100% sensitivity (29/29) and 91.7% specificity (11/12) when compared to the semi-nested PCR. In field validation using human fecal samples, the fluorescent RPA assay showed a sensitivity of 90.0% (36/40) and a specificity of 91.1% (225/247) compared to the Kato-Katz. And the sensitivity of the fluorescent RPA method compared to the semi-nested PCR method was 100% (34/34), while the specificity was 90.5% (229/252). The fluorescent RPA assay presents a rapid and dependable method for detecting N. americanus in fecal samples. Its high sensitivity and specificity provide significant utility for field surveillance and early identification of N. americanus infections. This advancement could facilitate the rapid molecular diagnosis of N. americanus disease in hookworm-endemic regions.

Characteristics and immunoprotective functions of three cysteine proteases from Clonorchis sinensis.

Cysteine proteases from Clonorchis sinensis, including various proteins, are essential for its pathogenicity and serve as potential vaccine candidates. This study assesses the protective effects of three C. sinensis cysteine proteases (CsCP1-3). Mice immunized with recombinant CsCP1-3 and adjuvants were subsequently infected with C. sinensis metacercariae after three immunization rounds. Liver damage was evaluated through hematoxylin and eosin (H&E), Masson's trichrome, and immunohistochemical analyses. The levels of IgG1, IgG2a antibodies, and cytokines (IFN-g, IL-2, IL-4, and IL-10) were quantified by enzyme-linked immunosorbent assay (ELISA). RT-qPCR revealed that CsCP1-2 exhibited the highest expression in newly encysted larvae (NEL), while CsCP3 was predominantly expressed in adult stages. Immunohistochemical localization confirmed that CsCP1-3 are present in the eggshells, syncytial layers of metacercariae, NEL cuticle, and adult intestines. Histological and immunohistochemical analysis demonstrated that the rCsCP1-3-immunized group displayed reduced liver inflammation and biliary fibrosis compared to the control group. The rCsCP1-3 induced a progressive increase in specific IgG1 and IgG2a antibody titers by the second week post-immunization. In the CsCP1-2 group, cytokines IFN-g, IL-2, IL-4, and IL-10 were elevated relative to the control, with particularly high levels of IFN-g and IL-10 in CsCP1, indicating a strong mixed Th1/Th2 immune response. In contrast, the CsCP3 immunization group exhibited a transient increase in cytokines (IFN-g, IL-2, IL-4, and IL-10) three days postinfection, which subsided after one to two weeks. These findings suggest that CsCP1-3 elicit robust antibody and cellular immune responses, mitigating liver damage caused by C. sinensis infection. CsCP1, in particular, induces a potent mixed Th1/Th2 response, positioning it as a promising vaccine candidate.

Carcinogenic parasites: Insights into the epidemiology and possible mechanisms of cancer.

Naturally, a wide range of genetic and environmental variables predominate, such as bacterial, viral and parasite infective entities that have been identified as carcinogenic bioagents. Many helminth and protozoan parasitic diseases are liable to cause human cancer. Conveniently, three trematode parasites viz. Schistosoma haematobium, Opisthorchis viverrini and Clonorchis sinensis have been reported to be intrinsically linked with human cancer. Similar studies for other parasitic infections are still imprecise and need further validation. Plasmodium falciparum is known to cause holoendemic Burkitt lymphoma despite non-carcinogenic role of malaria. This review is endowed with a coupled correlation and underlying mechanisms by which parasitic infections lead to carcinogenicity. An empirical documentation covering the prevalence and incidence of viral, bacterial and parasitic carcinogenicity is illustrated in this article. Moreover, some probable diagnostic and treatment procedures for parasitic carcinogenicity are also summarized. A detailed account of various mutational and genetic changes that lead to carcinogenesis via different pathways is appended in this article.

Clonorchis sinensis excretory/secretory proteins ameliorate inflammation in rheumatoid arthritis and ankylosing spondylitis

BackgroundWe aimed to investigate whether substances secreted by Clonorchis sinensis excretory/secretory protein (CS-ESP) have an effect on the inflammation of rheumatoid arthritis (RA) and ankylosing spondylitis (AS) and to identify specific peptides through related proteomic analysis to determine which proteins exhibit anti-inflammatory effects more specifically.MethodsPeripheral blood mononuclear cells (PBMCs) were obtained from healthy controls (HCs), RA and AS patients. Cytotoxicity of CS-ESP at different doses was assessed by MTS and flow cytometry before performing experiments. Inflammatory cytokine producing cells were analyzed using flow cytometry. To determine the effect of CS-ESP in an arthritis mouse model, 8-week-old SKG mice were injected intraperitoneally with curdlan and treated with CS-ESP; body weight and paw swelling were checked twice a week. Inflammation was evaluated using immunohistochemistry. We conducted proteomic analysis on CS-ESP and identified specific Cs-GT and Cs-Severin proteins. In vitro effect of coculture with Cs-GT and Cs-Severin was determined by inflammatory cytokine measurements.ResultTreatment with CS-ESP resulted in no reduced cell viability of PBMCs. In experiments culturing PBMCs, the frequencies of IL-17A and GM-CSF producing cells were significantly reduced after CS-ESP treatment. In the SKG mouse model, CS-ESP treatment significantly suppressed clinical score, arthritis and enthesitis. Treatment with Cs-GT and Cs-Severin resulted in no reduced cell viability of HC PBMCs. After Cs-GT and Cs-Severin treatment of HC PBMC, the frequencies of IL-17A and GM-CSF producing cells were significantly reduced.ConclusionsWe provide evidence showing that CS-ESP, Cs-GT and Cs-Severin can ameliorate clinical signs and cytokine derangements in AS.Graphical

Epidemiology and risk factors of Clonorchis sinensis infection in the mountainous areas of Longsheng County, Guangxi: insights from automated machine learning

Clonorchis sinensis (C. sinensis) is mainly prevalent in Northeast and South China, with Guangxi being the most severely affected region. This study aimed to evaluate the prevalence and identify the risk factors of C. sinensis infection in Longsheng County, a mountainous area in northern Guangxi. In 2023, a comprehensive study was conducted in Longsheng County, utilizing longstanding inhabitants as study participants. Questionnaires were employed to gather data on fish consumption, awareness of C. sinensis, and residential coordinates, while fecal examinations were utilized to identify C. sinensis infection. Important risk factors for the C. sinensis infection were identified through the development of individual infection risk models using automated machine learning techniques. A total of 740 fecal samples were collected, revealing an overall C. sinensis infection rate of 69.59%. The gradient boosting machine (GBM) was the most accurate predictor with an area under the precision-recall Curve (AUPRC) of 0.997. The model identified years of raw fresh fish consumption, frequency of raw fresh fish consumption, elevation, and water distance as the top four predictors of C. sinensis infection risk. In conclusion, our study has revealed a high infection rate of C. sinensis in the mountainous areas of Longsheng County, with adults, men, and farmers particularly susceptible to both high incidence and infection severity. We developed a high-performance predictive model for individual C. sinensis infection within the county, identifying the key risk factors for local infections. These findings offer valuable guidance for the control and prevention of clonorchiasis.

Clonorchis sinensis Infection prevents DSS-induced Colitis Via Lithocholic Acid in a Gut Microbiota-Dependent Manner.

Increasing evidence demonstrates that helminth and its components can ameliorate ulcerative colitis. Clonorchis sinensis (C. sinensis) is a kind of helminth that dwells in the bile ducts for many years, but the roles and underlying mechanisms of C. sinensis-induced protection from colitis are not elucidated. In the present study, the mice were infected with 50 C. sinensis metacercariae and further administrated with 4% Dextran Sodium Sulfate (DSS) in drinking water for 7days on days 49 post-infection. The disease severity and the integrity of gut barriers were evaluated. Gut microbiota was measured using 16sRNA sequencing, and bile acids in the colon were detected by Liquid Chromatography Mass Spectrometry (LC/MS). The Co-housing approach or microbiota deletion with additional supplies of secondary bile acids (SBAs) was employed to investigate the roles of gut microbiota in the protection from colitis. C. sinensis infection moderated the dysbiosis of the intestinal microbiota and increased the levels of SBAs and bile acid receptor Takeda G protein-coupled receptor 5 (TGR5), which finally benefited anti-inflammation and ameliorated the severity of DSS-induced colitis. Co-housing with C. sinensis-infected mice, and non-infected mice with colitis also showed an increase of TGR5, decreased pro-inflammatory cytokines, and a reduction in the severity of colitis, compared to those mice suffering from colitis without co-housing. Furthermore, C. sinensis-induced protective effects on colitis were attenuated by microbiota deletion, while SBAs (lithocholic acid, LCA) supplementation reversed the colitis. The present study demonstrates that C. sinensis infection ameliorates DSS-induced ulcerative colitis in mice, which is dependent on gut microbiota-associated SBAs.

The association between Clonorchis sinensis seropositivity and hepatocellular carcinoma in an endemic area: a study in Guangxi, China

BackgroundChronic infection with Clonorchis sinensis (C.sinensis) has been associated swith the development of intrahepatic cholangiocarcinoma (ICC); however, the relationship between C.sinensis and hepatocellular carcinoma (HCC) remains uncertain.MethodsThis study examined 120 patients with liver cancer in the clonorchiasis endemic area of Hengzhou, Guangxi, China. The type of cancer, the differentiation grade according to Edmondson Steiner’s classification, and the pathological characteristics of HCC were determined through postoperative tissue biopsy. C.sinensis infection was detected by measuring serum specific IgG antibody, and hepatitis B virus (HBV) infection was determined by detecting serum HBsAg and HBV DNA in HCC tissues. The C.sinensis infection rates in control groups were drawn from the local general population based on previous surveys. The association between C.sinensis infection and HCC was analyzed by comparing the differences in C.sinensis infection rates between the two groups.ResultsOf the patients evaluated, 98 (81.7%) had HCC, 21 (17.5%) had ICC, and 1 (0.8%) had comorbidity of HCC/ICC. Among the HCC patients, 24 (24.5%) were solely infected with HBV, 71 (72.4%) were C. sinensis seropositive, and 3 (3.1%) showed no evidence of infection. C. sinensis seropositive rates in HCC patients are much higher than in general outpatient and non-liver cancer inpatients (χ2 = 141.92, p < 0.001), as well as in the local residents (χ2 = 82.61/21.38, p < 0.001). There were no significant differences in the pathological type, differentiation grade, and lesion composition between the tumor associated with C.sinensis/HBV mono- and co-infection (p > 0.05). Among the patients with C.sinensis-related HCC, 8 (8.2%) were solely C.sinensis seropositive, while 63 (64.3%) were co-infected with HBV. Infection with C. sinensis and HBV has a significant impact on the pathological types of liver cancer (χ2 = 22.86, p < 0.001).ConclusionsThese findings indicate that HCC still accounts for the majority of liver cancer in this region. In addition to being most commonly related with HBV infection, HCC may also be related to C. sinensis infection. Co-infection of C. sinensis and HBV may enhance the development of HCC in this area.Clinical trialNot applicable.

The microRNA landscape and regulatory network in Clonorchis sinensis-infected hepatocellular carcinoma: implications for tumor progression

BackgroundHepatocellular carcinoma (HCC) is a leading cause of cancer mortality globally, and its progression is associated with various factors, including parasitic infections such as Clonorchis sinensis (C. sinensis). Although C. sinensis infection has been implicated in HCC, the molecular mechanisms, particularly the role of microRNAs (miRNAs), remain poorly understood. This study aims to fill this gap by investigating the miRNA expression profiles in C. sinensis+ and C. sinensis− HCC tissues.MethodWe performed miRNA sequencing on HCC tissues from C. sinensis+ and C. sinensis− patients, followed by bioinformatics analyses to identify differentially expressed miRNAs (DEMs) and their target genes. Gene Ontology (GO) enrichment analysis was conducted to explore relevant biological processes, while a competitive endogenous RNA (ceRNA) network was constructed to investigate the interactions among miRNAs, long noncoding RNAs (lncRNAs), and messenger RNAs (mRNAs). Additionally, we performed survival analysis using Gene Expression Profiling Interactive Analysis 2 (GEPIA2) based on the The Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA-LIHC) cohort and assessed the clinical relevance of DEMs. Key miRNAs identified from this analysis were further validated through quantitative real‑time polymerase chain reaction (qRT-PCR) assays to confirm their expression in MHCC97H.ResultsOur research identified significant miRNA dysregulation in C. sinensis+ HCC tumors compared with C. sinensis− HCC tumors. Notably, miR-143-3p, miR-10a-5p, and miR-100-5p were upregulated in C. sinensis+ HCC, contributing to immune responses and tumor progression, while let-7 family members and miR-221-3p were downregulated, affecting metabolic pathways. GO enrichment analysis highlighted the involvement of developmental processes, immune system regulation, and metabolic reprogramming in C. sinensis+ HCC. The construction of a ceRNA network revealed key interactions between miRNAs, lncRNAs, and mRNAs in C. sinensis+ HCC, suggesting regulatory mechanisms that could be potential therapeutic targets. Additionally, validation through qRT-PCR confirmed these findings, highlighting miRNA dysregulation as a critical factor in C. sinensis+ HCC progression.ConclusionsThis study provides novel insights into the role of miRNAs in C. sinensis-infected HCC progression. The findings highlight the critical role of miRNA dysregulation in the progression of C. sinensis-associated HCC, emphasizing the potential for therapeutic interventions targeting these molecular alterations in affected patients.Graphical

Correlation analysis of the impact of Clonorchis sinensis juvenile on gut microbiota and transcriptome in mice.

This study highlighted the impact of C. sinensis juvenile infection on the gut microbiota and transcriptome of BALB/c mice. It induced liver inflammation, promoted intestinal villi growth, and altered goblet cell numbers. The infection also disrupted the diversity and structure of gut microbiota, particularly affecting beneficial bacteria. Transcriptome analysis revealed increased expression of genes related to immune response and detoxification processes. Important pathways affected included circadian rhythm, glutathione metabolism, and glycolysis/gluconeogenesis. Notable genes implicated included Igkv12-41, Mcpt2, Arntl, Npas2, Cry1, and Gsta1. Bacteroides_sartorii emerged as a potential key regulator in this interaction.

Search for and functional annotation of multi-domain PLA2 family proteins in flatworms.

The phospholipase A2 (PLA2) is a superfamily of hydrolases that catalyze the hydrolysis of phospholipids and play a key role in many molecular processes in the cells and the organism as a whole. This family consists of 16 groups divided into six main types. PLA2 were first isolated from venom toxins and porcine pancreatic juice. The study of these enzymes is currently of great interest, since it has been shown that a number of PLA2 are involved in the processes of carcinogenesis. PLA2 enzymes were characterized in detail in model organisms and humans. However, their presence and functional role in non-model organisms is poorly understood. Such poorly studied taxa include flatworms, a number of species of which are human parasites. Several PLA2 genes have previously been characterized in parasitic flatworms and their possible role in parasite-host interaction has been shown. However, no systematic identification of the PLA2 genes in this taxon has been carried out. The paper provides a search for and a comparative analysis of PLA2 sequences encoded in the genomes of flatworms. 44 species represented by two free-living and 42 parasitic organisms were studied. The analysis was based on identification of orthologous groups of protein-coding genes, taking into account the domain structure of proteins. In flatworms, 12 of the 13 known types of animal A2 phospholipases were found, represented by 11 orthologous groups. Some phospholipases of several types fell into one orthologous group, some types split into several orthogroups in accordance with their domain structure. It has been shown that phospholipases A2 of the calcium-independent type, platelet-activating phospholipases from group G8 and lysosomal phospholipases from group G15 are represented in all large taxa of flatworms and the vast majority of the species studied by us. In free-living flatworms PLA2 genes have multiple copies. In parasitic flatworms, on the contrary, loss of genes occur specifically in individual taxa specifically for groups or subfamilies of PLAs. An orthologous group of secreted phospholipases has been identified, which is represented only in Digenea and this family has undergone duplications in the genomes of opisthorchids. Interestingly, a number of experimental studies have previously shown the effect of Clonorchis sinensis proteins of this orthogroup on the cancer transformation of host cells. Our results made it possible for the first time to systematically identify PLA2 sequences in flatworms, and demonstrated that their evolution is subject to gene loss processes characteristic of parasite genomes in general. In addition, our analysis allowed us to identify taxon-specific processes of duplication and loss of PLA2 genes in parasitic organisms, which may be associated with the processes of their interaction with the host organism.

Epidemiological characteristics and precise prophylaxis and control of HBV-associated primary liver cancer

Primary liver cancer (PLC), which includes hepatocellular carcinoma (HCC, 93% in China; 75%-80% worldwide), intrahepatic cholangiocarcinoma (ICC, 4.3% in China; 10%-15% worldwide), and combined hepatocellular cholangiocarcinoma (CHC, 1.6% in China), is a global disease that brings a heavy burden to the world and the number of incidence cases is on the rise. Chronic liver injury caused by factors such as exposure to aflatoxin B1, infection with Clonorchis sinensis, heavy alcohol consumption, chronic infection with hepatitis C virus (HCV), and metabolic syndrome are all known risk factors for PLC. Notably, chronic infection with hepatitis B virus (HBV) is the major risk factor for HCC. Globally, PLC risk factors are changing from infectious causes to metabolic factors. Here, we update the mechanisms of HBV-related HCC (HBV-HCC) development, especially the effect of HBV evolution on the development of HCC. The HBV mutations, viral load, and HBV integration, together with parameters of poor liver function, are key components to define the highest-risk population of HBV-HCC. Antiviral therapy has been proven to be effective for the prevention of HBV-HCC in the highest-risk population. Non-invasive imaging combined with key markers is economical and convenient for screening early PLC. Surgical resection and liver transplantation are therapeutic options for HCC; however, postoperative recurrence reaches 70% in five years. Targeted therapy, immunotherapy, and radiotherapy can improve the survival of PLC. Active prophylaxes, including HBV vaccination, antiviral treatment, improving lifestyle to decrease chronic inflammation, and surveillance, are cost-effective in decreasing the disease burden of PLC.

Clonorchis sinensis Promotes Intrahepatic Cholangiocarcinoma Progression by Activating FASN-Mediated Fatty Acid Metabolism.

Clonorchis sinensis infection is an important risk factor for intrahepatic cholangiocarcinoma (ICC). C. sinensis positive (C.s+) ICC patients had much shorter overall survival (OS) compared with C. sinensis negative (C.s-) group. This study aims to explore the impact and underlying mechanism of C. sinensis infection on ICC progression. In this study, ICC patients underwent surgery from two medical centers enrolled. RNA sequencing was used to determine the downstream activated pathways and genes. Furthermore, we demonstrated the potential mechanism of C. sinensis infection in promoting ICC progression through invitro co culture systems and two animal models. Through RNA sequencing, we found fatty acid metabolism and the expression of fatty acid synthase (FASN), a key enzyme catalyzing long-chain fatty acid synthesis, were significantly elevated in C.s+ ICCs. Then, we found excretory/secretory products (ESPs) secreted by C. sinensis could significantly upregulate the expression of transcription factor E2F1, thereby promoting FASN expression and fatty acid synthesis in tumor cells, which ultimately accelerating tumor progression. However, the promotive effect disappeared when FASN was knocked down. Meanwhile, ESPs could promote tumor growth, increasing FASN expression and free fatty acid level in both subcutaneous and orthotopic mouse models. This study indicates that C. sinensis infection could upregulate the level of FASN and activate fatty acid synthesis pathway, thereby accelerating ICC progression. This provides a new insight for the clinical treatment of ICC with C. sinensis infection.

Serum proteomic and metabolomic profiling of hepatocellular carcinoma patients co-infected with Clonorchis sinensis.

Clonorchis sinensis (C. sinensis) infection is a significant risk factor for hepatocellular carcinoma (HCC), yet its underlying mechanisms remain poorly understood. This study aimed to investigate the impact of C. sinensis infection on the serum proteomic and metabolomic profiling of HCC patients, focusing on the potential mechanisms. A retrospective clinical analysis was conducted on 1121 HCC patients, comparing those with and without C. sinensis infection. The influence of C. sinensis on serum proteome and metabolome in HCC was further assessed. C. sinensis infection correlated with a younger age at cancer onset, male predominance, advanced cancer stage, liver cirrhosis, and microvascular invasion in HCC patients. It also associated with shorter overall survival (OS) and recurrence-free survival (RFS). The levels of blood lipids (e.g., APO-A, HDL-C, and TG) were significantly altered after C. sinensis infection. Proteomic and metabolomic analyses revealed metabolic reprogramming caused by C. sinensis, with excessive depletion of argininosuccinate synthase (ASS) and D-glucose as potential factors in C. sinensis-associated HCC malignancy. Key molecules ILF2, CNN2, OLFM4, NOTCH3, and LysoPA were implicated in HCC progression. Furthermore, C. sinensis triggered inflammation, insulin resistance, and pro-tumor immune escape, and exacerbated the complication of degenerative diseases. This study not only provides compelling evidence for elucidating the mechanisms underlying C. sinensis-mediated HCC development but also identifies potential therapeutic targets for HCC patients co-infected with C. sinensis.

Assessment of the application of the FA280—a fully automated fecal analyzer for diagnosing clonorchiasis: a mixed-method study

BackgroundClonorchiasis is an important foodborne parasitic disease in China caused by Clonorchis sinensis. Accurate and rapid diagnosis of this disease is vital for treatment and control. Traditional fecal examination methods, such as the Kato-Katz (KK) method, are labor-intensive, time-consuming, and have limited acceptance. The FA280, an advanced automated fecal analyzer, increases efficiency while significantly reducing labor load. This study aims to evaluate its performance, applicability, and scalability in clonorchiasis diagnosis to explore its potential application in the future.MethodsA mixed-methods study integrating both quantitative and qualitative approaches was conducted. The quantitative component consisted of a cross-sectional survey in Xinhui District, Guangdong, China, to evaluate the diagnostic performance of the FA280. The positive rate and agreement between the FA280 and the KK method were evaluated using McNemar’s test. Additionally, Pearson’s Chi-square test was used to analyze the consistency of positive results between the two methods across various eggs per gram (EPG) groups under different cut-off values. The qualitative component included semi-structured individual interviews with medical staff and institutional administrators to examine the FA280’s applicability and potential for broader adoption, with thematic analysis of the data.ResultsIn the quantitative study of 1000 participants, both the FA280 and KK methods detected clonorchiasis with a positive rate of 10.0%, achieving 96.8% agreement and showing no significant difference (P > 0.999). The kappa value was 0.82 (95% confidence interval: 0.76–0.88), indicating a strong agreement between the methods. The agreement rate for positive results between the two methods was significantly higher in the high infection intensity group compared to the low infection intensity group (P < 0.05). The qualitative study, which involved interviews with three medical staff and two administrators revealed that the FA280 outperformed the KK method in testing procedures, detection results, and user acceptance. The benefits, challenges, and suggestions of FA280 promotion were also emphasized.ConclusionsThis study demonstrated the FA280’s application value in clonorchiasis diagnosis by assessing its detection performance, applicability, and scalability. These findings contribute to the future prevention and control of the disease.Graphical

Clonorchis sinensis-infected hepatocellular carcinoma exhibits distinct tumor microenvironment and molecular features.

Clonorchis sinensis (Cs)-infected hepatocellular carcinoma (HCC) patients have a poorer prognosis than non-Cs-infected HCCs. However, the molecular mechanisms of Cs-infected HCC remain unclear. To address this, this study aims to uncover the tumor microenvironment and molecular features that may contribute to these poor outcomes. The research involved bulk RNA sequencing of paired tumor and adjacent tissue samples from 10 Cs + HCC and 10 Cs - HCC patients. Differentially expressed genes were identified, followed by enrichment analyses to reveal functional changes. Survival analysis of the top 10 up- and down-regulated genes in Cs + HCC tumors was performed using TCGA database. Additionally, clinical data from 1,461 HCC patients were retrospectively analyzed to assess the impact of Cs infection on microvascular invasion and metastasis rates. In vitro assays were also conducted using Cs excretory/secretory products (CsESPs) to examine their effect on HCC cells and HUVECs. We identified 785 up-regulated and 675 down-regulated genes in Cs + HCC tumors compared to Cs - HCC tumors, enriched in pathways related to extracellular matrix remodeling and immunosuppression. Survival analysis revealed that the top 10 up-regulated genes are associated with HCC poor prognosis. Clinical data from 1,461 HCC patients showed Cs infection increased microvascular invasion and metastasis rates. In vitro, CsESPs products enhanced migration and invasion in HCC cells and promoted tube formation in human umbilical vein endothelial cells. This study provides novel insights into the molecular landscape of Cs-infected HCC and underscores the Cs infection's role in enhancing tumor migration, invasion and angiogenesis. The findings contribute to the understanding of parasitic infections in cancer progression and suggest potential prognostic markers for Cs + HCC.