Serum proteomic profiling in patients with advanced Schistosoma japonicum-induced hepatic fibrosis.

BACKGROUND: Schistosoma japonicum is a parasitic flatworm that is the aetiological agent of human schistosomiasis, an important cause of hepatic fibrosis. Schistosomiasis-induced hepatic fibrosis is a consequence of the highly fibrogenic nature of egg-induced granulomatous lesions, which are the main pathogenic features of schistosomiasis. Although global awareness of the association between schistosomiasis-induced hepatic fibrosis and S. japonicum infection is increasing, little is known about the molecular differences associated with rapid progression to schistosomiasis in cirrhotic patients. METHODS: We systematically used data-independent acquisition (DIA)-based liquid chromatography-mass spectrometry to identify differentially expressed proteins in serum samples from patients with advanced S. japonicum-induced hepatic fibrosis. RESULTS: Our analysis identified 1144 proteins, among which 66 were differentially expressed between the healthy control group and the group of patients with advanced S. japonicum-induced hepatic fibrosis stage F2 (SHF-F2) and 214 were differentially expressed between the SHF-F2 and SHF-F4 groups (up- or downregulation of at least 1.5-fold in serum samples). The results also indicated that two selected proteins (C1QA and CFD) are potential biomarkers for distinguishing between patients with SHF-F2 and those with SHF-F4 due to S. japonicum infection. CONCLUSIONS: We provide here the first global proteomic profile of serum samples from patients with advanced S. japonicum-induced hepatic fibrosis. The proteins C1QA and CFD are potential diagnostic markers for patients with SHF-F2 and SHF-F4 due to S. japonicum infection, although further large-scale studies are needed. Our DIA-based quantitative proteomic analysis revealed molecular differences among individuals at different stages of advanced S. japonicum-induced hepatic fibrosis and may provide fundamental information for further detailed investigations.

Evaluation of transient elastography in assessing liver fibrosis in patients with advanced schistosomiasis japonica.

Schistosomiasis remains an important public health issue. The presence and extent of liver fibrosis are associated with disease progression and prognosis. The study is aimed at exploring the value of liver stiffness measurement (LSM) by transient elastography in assessing liver fibrosis in patients with advanced schistosomiasis japonica. Seventy-three patients were consecutively recruited for the purpose of this study. The correlation between noninvasive parameters and histological fibrosis stages was analyzed and an area under receiver operating characteristic curve (AUROC) was used to assess diagnostic efficacy. Our results demonstrated that there are significant differences between LSM values of patients with different stages of fibrosis (F1 vs. F2, F2 vs. F3 and F3 vs. F4, P<0.01). The AUROC values of LSM in detecting significant fibrosis (F≥2), advanced fibrosis (F≥3) and cirrhosis (F=4) were 0.96, 0.90, and 0.92 respectively. The optimal cut-off LSM values were 8.0kPa, 9.5kPa, and 18.0kPa for significant fibrosis, advanced fibrosis and cirrhosis. Based on differences between AUROC values, LSM was proven to be superior to several serum models in detecting advanced fibrosis and cirrhosis. In conclusion, our study demonstrates that LSM is a reliable parameter for assessing risk of liver fibrosis in patients with advanced schistosomiasis japonica.

R-spondin 2 promotes proliferation and migration via the Wnt/ß-catenin pathway in human hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a leading cause of malignant disease-associated mortality, particularly in China. The RSPO2 (R-spondin 2) gene is evolutionarily conserved in vertebrates and is involved in developmental and physiological processes. Importantly, RSPO2 has been reported to be associated with colon cancer and potentiate the Wnt/ß-catenin signaling pathway. In the present study, enhanced expression of RSPO2 in HCC was observed using tissue microarray. Similarly, the expression level of RSPO2 was higher in HepG2, Huh7 and Hep3B cells but lower in Bel7404 and QGY7703 cells compared with human normal QSG7701 liver cells. Subsequently, gain-of-function studies indicated that RSPO2 promotes the proliferation and migration of QGY7703 cells based on lentivirus-based gene delivery. Furthermore, it was revealed that p21 and leptin, rather than vascular endothelial growth factor-A, are involved in the function of RSPO2 in QGY7703 cells. Particularly, the signal transducer and activator of transcription 3 (STAT3) and Wnt/ß-catenin signaling pathways are involved in this process. Overexpression of RSPO2 resulted in the elevated expression of phosphorylated STAT3, ß-catenin and c-Myc. Therefore, the present study is beneficial to the understanding of RSPO2-involved liver cancer transformation and drug discovery.

RSPOs facilitated HSC activation and promoted hepatic fibrogenesis.

Roof plate-specific spondin (RSPO) proteins are potent Wnt pathway agonists and involve in a broad range of developmental and physiological processes. This study investigated the activities and mechanisms of RSPOs in liver fibrogenesis, especially in hepatic stellate cell (HSC) activation. HSC activation was assessed by fibrosis biomarker (α-smooth muscle actin and Collagen-I), phenotypic change (accumulation of lipid droplets), and increased proliferation. Similarly, Wnt pathway activity was evaluated by the expression of nuclear ß-catenin and T cell-specific transcription factors (TCF) activity. We found RSPOs were overexpressed in human fibrotic liver tissue and the expressions were correlated with liver fibrosis stages. In vitro studies showed RSPOs level increased during HSC activation, and stimuli with RSPOs enhanced Wnt pathway activity and promoted HSC activation subsequently. Furthermore, in vivo experiments demonstrated that the knockdown of RSPOs suppressed both Wnt pathway activity and HSC activation. Interestingly, the inhibitor of the Wnt signaling pathway Dickkopf1 impairs RSPOs effects on HSCs. Taken together, our results revealed that RSPOs facilitated HSC activation and promote liver fibrogenesis by enhancing the Wnt pathway.

[Role of R-Spondin1 in the activation of hepatic stellate cells].

OBJECTIVE: To investigate the role of R-Spondinl in the activation of hepatic stellate cells (HSCs). METHODS: Twenty-four healthy male Kunming mice were randomly divided into the following two groups:fibrosis model group (n=16) and control group (n=8). Hepatic fibrosis was induced by subcutaneous injections of CC14 (20% in olive oil) at a dose of 5 ml/kg twice per week. After 10 weeks, the animals were sacrificed by CO(2) over-exposure and liver tissues were harvested.The protein and mRNA levels of R-Spondin1, alphat-SMA,and collagen I were examined by Western blot assay and real-time PCR respectively. Additionally,HSCs were isolated from the mice liver tissues to examine the time-series expression changes of R-Spondinl, alpha-SMA, and nuclear beta-catenin.TCF activity was analyzed by luciferase reporter assay.Moreover,HSCs were cocultured with recombinant R-Spondin1 and DKK1 to evaluate dose-response. RESULTS: R-Spondinl expression was significantly higher in the fibrosis model group than in the control group (protein level:3.16 ± 0.18 vs. 0.99 ± 0.16, t =13.31, P < 0.01; mRNA level:4.36 ± 0.26 vs. 0.98 ± 0.12, t =21.46, P < 0.01).The culture-activated mouse HSCs showed up-regulated TCF activity (5.33 ± 0.34 vs. non-activated: 1.03 ± 0.09, t =20.93, P < 0.01), nuclear beta-catenin expression (4.47 ± 0.21 vs. 0.97 ± 0.14, t =25.25, P < 0.01), and R-Spondin1 expression (protein level: 4.54 ± 0.18 vs. 1.04 ± 0.12, t =31.17, P < 0.01; mRNA level:5.13 ± 0.15 vs. 1.01 ± 0.16, t=38.06, P < 0.01). Exogenous stimulation of freshly isolated mouse HSCs with recombinant R-Spondin1 induced a dose-dependent increase in both TCF activity and the expression of nuclear beta-catenin and alphat-SMA. DKK1 down-regulated activities of factors in the WNT signaling pathway and repressed activation of HSCs. Conclusion R-Spondin1 may promote HSC activation by enhancing the canonical WNT signaling pathway.

Clinical and epidemiological characteristics of adult hand, foot, and mouth disease in northern Zhejiang, China, May 2008-November 2013.

BACKGROUND: Hand, foot, and mouth disease (HFMD) is an infectious disease typically caused by enterovirus 71 (EV71) and Coxsackievirus A16. The incidence of HFMD appears to be increasing across the Asia Pacific region, with deaths occurring predominantly among children. Therefore, most HFMD reports focus on children and few have studied HFMD in adults. However, more adult HFMD cases may be seen in the foreseeable future as a result of global warming, continued viral evolution, and an increase in traveling. Thus, this study investigated the clinical and epidemiological characteristics of adult HFMD. METHODS: Case data of 49 adult HFMD patients who attended The First Affiliated Hospital of Jiaxing College, China from May 2008 to November 2013 were obtained. Socio-demographic data were collected through follow-up phone calls. Throat swab specimens were tested for enterovirus by quantitative reverse transcription-polymerase chain reaction and further confirmed by virus isolation assay. For 10 patients infected with EV71, the gene encoding the EV71 VP1 protein was sequenced and analyzed. Data from 8,354 child HFMD patients and 49 adult patients in the fever clinic of The First Affiliated Hospital of Jiaxing College during the same period were collected for comparison. RESULTS: This study revealed that close contact with HFMD patients and poor personal hygiene consciousness were risk factors for adult HFMD. This study also found that EV71 subgenotype C4a was the most common pathogen associated with adult HFMD in this area. Furthermore, this study demonstrated several unique epidemiological characteristics of adult HFMD compared to child HFMD, such as the geographic and gender distribution of adult HFMD patients and HFMD seasonality. CONCLUSIONS: The findings in this study showed the potential threat of adult HFMD.

R-spondin2 activates hepatic stellate cells and promotes liver fibrosis.

BACKGROUND: The development of liver fibrosis is the fundamental stage toward a number of mortal complications of liver diseases, including cirrhosis and hepatocellular carcinoma. Canonical Wnt pathway is crucial in diverse biological processes and mediates the progression and regression of liver fibrosis. As a potent Wnt pathway agonist, the role of roof plate-specific spondin-2 (R-Spondin2) in the hepatic fibrosis has not been well elucidated. AIMS: The purpose of this study was to investigate whether R-Spondin2 contributes to hepatic stellate cells (HSCs) activation, the key event in liver fibrogenesis. METHODS: Human liver tissues, hepatic fibrosis mouse model, and freshly isolated mice HSCs were used. Protein expression and transcriptional level were analyzed by Western-blot assays and real-time PCR, respectively. Exogenous stimulation with recombinant R-Spondin2 and knockdown of R-Spondin2 were performed to investigate functionality. Nuclear ß-catenin level and T cell-specific transcription factors activity were analyzed, and HSC proliferation was tested by MTT assay. RESULTS: Overexpression of R-Spondin2 was observed in both human fibrotic liver tissues and hepatic fibrosis mouse model. Exogenous stimulation with R-Spondin2 in the freshly isolated mice HSCs induced a dose-dependent increase in Wnt pathway activities, HSC proliferation, and the expression of α-smooth muscle actin (α-SMA) and Collagen I. Additionally, Wnt pathway activities, HSC proliferation, and the expressions of α-SMA and Collagen I decreased in the R-Spondin2 knockdown HSCs. CONCLUSIONS: These findings suggest that R-Spondin2 may promote HSC activation by enhancing the canonical Wnt pathway.