Impact of anti-<i>C</i>. <i>sinensis</i> treatment on antiviral therapies in co-infected patients.

<p>Elevated liver transaminases (including AST, ALT and TB) and HBV DNA copies in the plasma of co-infected patients were detected after receiving combination treatment ETV and PZQ or not. (A) AST and (B) ALT and (C) TB were measured in plasma. (D) HBV DNA levels were measured by RT-PCR. Symbols show individual measurements within the patient groups, and the graphs show the means ± SD. Asterisks indicate statistically significant differences between NONPZQ and PZQ groups, as measured by paired, two-tailed Student's t-test (*<i>p</i> <0.05, ** <i>p</i> <0.01)</p

Clinical characteristics of the study cohort.

<p>Clinical characteristics of the study cohort.</p

Primer sequences for quantitative real-time PCR.

<p>Primer sequences for quantitative real-time PCR.</p

Detection of HBV-DNA copies in PBMC culture supernatant.

<p>HBV DNA was detected by FQ-PCR in the supernatant of the PBMCs incubated with mixtures of ESP and HBV positive serum or HBV positive serum only, or ESPs only. Medium alone served as a control. Asterisks indicate statistically significant differences between HBV positive sera only and mixtures of HBV positive sera and ESPs, as measured by paired, two-tailed Student's t-test (** <i>p</i> <0.01).</p

mRNA levels of Th1 and Th2 cytokines secreted by stimulated PBMCs.

<p>Total RNAs from PBMCs stimulated by mixtures of ESP and HBV positive serum, HBV positive serum only, or ESPs only were extracted for reverse transcription using cytokine gene-specific primers for Th1 cytokine (A) including IL-2 and IFN-γ and Th2 cytokines (B) including IL-4, IL-6 and IL-10 and human β-actin. The relative expression of each cytokine was detected by quantitative real-time RT-PCR and normalized relative to β-actin expression. Medium only served as a control. Data are shown as the mean ± SEM (*<i>p</i> <0.05, ** <i>p</i> <0.01, *** <i>p</i> <0.001).</p

Immunolocalization of <i>Cs</i>HK in <i>C</i>. <i>sinensis</i> and in liver from infected rats.

<p>Mouse anti-r<i>Cs</i>HK serum and anti-mouse IgG were applied as primary antibody and secondary antibody, respectively. Serum from pre-immune mice was employed as primary antibody for a negative control. Panels H, L, P, R, U, V, W, and X are negative controls. Panels B, D, F, H, J, L, N, P, and R are under fluorescence microscope and the same parts (panels A, C, E, G, I, K, M, O, and Q) are under white light. Panels B, D, and F, localization of <i>Cs</i>HK in adult worms; panel J, localization of <i>Cs</i>HK in metacercariae. Panels S and T, localization of <i>Cs</i>HK in intrahepatic bile ducts of a <i>C</i>. <i>sinensis</i> infected rat. In panels S, T, U, V, W, and X, peroxidase staining shows as a yellow/rust colored deposit and Mayer’s hematoxylin counterstains the nuclei in light purple. White arrows highlight the regions of intrahepatic bile duct tissue and the tissue that stained positive for <i>Cs</i>HK. Original magnification: × 50 for panels M, N, O, P, Q and R; × 100 for panels A, B, C, D, E, F, G, H, S, U, and W; × 400 for panels I, J, K, L, T, V, and X. Bar = 800 μm. v, vitellarium; e, egg; vs, ventral sucker; tg, tegument; i, intestine; u, uterus; ts, testicle; o, ovary; p, pharynx; s, spermatheca; l, lumen; w, within the cells; <i>Cs</i>, <i>C</i>. <i>sinensis</i>; BE, biliary epithelium.</p

Rat anti-r<i>Cs</i>HK serum affects <i>C</i>. <i>sinensis</i> adult survival in vitro.

<p>(A) The median survival of <i>C</i>. <i>sinensis</i> adults in the blank control group, 1:40 pre-immune serum group, 1:80 pre-immune serum group, 1:160 pre-immune serum group, 1:40 anti-r<i>Cs</i>HK serum group, 1:80 anti-r<i>Cs</i>HK serum group, and 1:160 anti-r<i>Cs</i>HK serum group was 15, 8, 8, 9, 2, 3, and 3 days, respectively. There was no significant difference in survival rates among pre-immune serum groups at any dilution (<i>p</i> > 0.05). Significant differences were observed in the survival rates among the other groups (<i>p</i> < 0.05). (B) The enzymatic activity of <i>Cs</i>HK in homogenate of parasites collected from each group at 1, 3, 5, and 6 days of incubation. The enzymatic activity of <i>Cs</i>HK in adult worms incubated in medium with different dilutions of anti-r<i>Cs</i>HK serum declined significantly in a dose- and time-dependent manner. (C) As a control, there was no obvious change of the enzymatic activity of <i>Cs</i>PLA₂ in the worms.</p

Effects of phosphate donors, effectors and EbSe on the enzyme kinetics of r<i>Cs</i>HK.

<p>The effect of 0~3 mM phosphate donors (ATP, CTP, GTP, ITP, TTP, and UTP) and fixed 3 mM glucose (A). The effect of 0~5 mM AMP (B), 0~10 mM PEP (C), 0~10 mM citrate (D), or 0~100 μM EbSe (E) and fixed 3 mM glucose with respect to ATP. The effect of 0~100 μM EbSe and fixed 3 mM ATP with respect to glucose (F).</p

Worm burden and EPG of rats in different groups.

<p>Results of analysis represent the mean ± SD, and the recovered worm numbers and EPG in groups were compared by Student’s <i>t</i>-test.</p><p>^a<i>p</i> > 0.05 and</p><p>^b<i>p</i> < 0.01 (compared with PBS group).</p><p>Worm burden and EPG of rats in different groups.</p

Substrate specificity of r<i>Cs</i>HK.

<p>^a from reference [<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003641#pntd.0003641.ref017" target="_blank">17</a>].</p><p>Substrate specificity of r<i>Cs</i>HK.</p