Effect of siRNA-mediated silencing of ZFP36 on cytokine production by T cells.

<p><b>A.</b> Extent of ZFP36 silencing in PBMCs from healthy donors (n = 4). Total PBMCs from healthy donors were transfected by nucleofection with non-specific siRNA (NC siRNA) or ZFP36-specific siRNA (ZFP36 siRNA). qPCR analysis was used to quantify the expression of ZFP36 (p<0.001). <b>B.</b> Cytokine profiles in the supernatant of transfected and stimulated (anti-CD3/CD28) PBMCs. <b>C.</b> Cytokine profiles in the supernatant of transfected and stimulated (CpG ODN2006) PBMCs. The expression levels (pg/ml) of cytokines that displayed significant in control and ZFP36-silenced PBMCs are shown. * p<0.05; ** p<0.01; *** p<0.001. Data were obtained using cells from 4 different donors.</p

Characteristics of CHB patients and healthy donors.

<p>ALT, alanine aminotransferase; AST, aspartate aminotransferase; DBIL, direct bilirubin; TBIL, total bilirubin.</p

Comparison of cytokines increased in the plasma of CHB patients and the supernatant of PBMCs transfected with ZFP36 siRNA and stimulated with anti-CD3/CD28 or CpG.

*<p>p<0.05;</p>**<p>p<0.01;</p>***<p>p<0.001.</p><p>ARE data were obtained from the ARE-Containing mRNA Database from <a href="http://brp.kfshrc.edu.sa/ARED/" target="_blank">http://brp.kfshrc.edu.sa/ARED/</a>. Bracketed numbers indicate references.</p

Downregulation of ZFP36 in T lymphocytes, monocytes, and total PBMCs of chronic HBV patients.

<p>ZFP36 mRNA expression was measured by qPCR in purified CD4⁺ T lymphocytes (<b>A</b>), CD8⁺ T lymphocytes (<b>B</b>), CD14⁺ monocytes (<b>C</b>), and total PBMCs (<b>D</b>) from chronic HBV patients (n = 13) and healthy donors (n = 9). ZFP36 expression in control cells was defined as 1.0. P values for all four groups were <0.01. <b>E.</b> Inverse correlation between ZFP36 expression in total PBMCs and ALT level (U/L) in chronic HBV patients (p = 0.011). <b>F.</b> Correlation between ZFP36 expression in total PBMCs and HBV DNA (p = 0.416), AST level (p = 0.056), and TBIL level (p = 0.770) in chronic HBV patients.</p

Plasma cytokine profiles of chronic HBV patients.

<p><b>A and B.</b> Luminex results show the expression levels of 25 cytokines in the plasma of healthy donors (HD) (n = 10) and chronic HBV patients (n = 16). The expression levels (pg/ml) of cytokines that displayed significant differences between healthy donors and HBV patients are shown in A, and the expression levels of cytokines that did not display significant differences are shown in B. * p<0.05; ** p<0.01; *** p<0.001. <b>C.</b> Correlation between IL-8 and ALT levels (p<0.05); IL-8 and AST levels (p<0.05); TNF-α and HBV DNA levels (p<0.001); and IL-1RA and HBV DNA levels (p<0.05) in chronic HBV patients.</p

Effect of siRNA-mediated silencing of ZFP36 on T cell activation, proliferation, and survival.

<p>PBMCs from healthy donors were transfected by nucleofection with non-specific siRNA (NC siRNA) or ZFP36-specific siRNA (ZFP36 siRNA). <b>A.</b> Expression of CD25 and CD69 in CD4⁺ and CD8⁺ T cells was measured 24 h after stimulation with anti-CD3/CD28. Dotted lines: un-stimulated cells; solid lines: stimulated cells; grey lines: NC siRNA-transfected cells; black lines: ZFP36 siRNA-transfected cells. <b>B.</b> CFSE dilution of labeled CD4⁺ and CD8⁺ T lymphocytes. Dotted lines: cells transfected with NC siRNA; solid lines: cells transfected with ZFP36 siRNA. <b>C.</b> Cell cycle status of ZFP36 siRNA-transfected T cells. Cells were stained with PI. Solid lines: cells transfected with NC siRNA; dotted lines: cells transfected with ZFP36 siRNA. <b>D.</b> Apoptotic rates of ZFP36 siRNA-transfected T cells as determined by Annexin V staining. Percentages represent the frequency of Annexin V⁺ cells within the CD4⁺ and CD8⁺ T cell populations of PBMCs transfected with NC siRNA or ZFP36 siRNA.</p

Will Sofosbuvir/Ledipasvir (Harvoni) Be Cost-Effective and Affordable for Chinese Patients Infected with Hepatitis C Virus? An Economic Analysis Using Real-World Data

<div><p>Background</p><p>Little is known on the cost-effectiveness of novel regimens for hepatitis C virus (HCV) compared with standard-of-care with pegylated interferon (pegIFN) and ribavirin (RBV) therapy in developing countries. We evaluated cost-effectiveness of sofosbuvir/ledipasvir for 12 weeks compared with a 48-week pegIFN-RBV regimen in Chinese patients with genotype 1b HCV infection by economic regions.</p><p>Methods</p><p>A decision analytic Markov model was developed to estimate quality-adjusted-life-years, lifetime cost of HCV infection and incremental cost-effectiveness ratios (ICERs). SVR rates and direct medical costs were obtained from real-world data. Parameter uncertainty was assessed by one-way and probabilistic sensitivity analyses. Threshold analysis was conducted to estimate the price which can make the regimen cost-effective and affordable.</p><p>Results</p><p>Sofosbuvir/ledipasvir was cost-effective in treatment-experienced patients with an ICER of US$21,612. It varied by economic regions. The probability of cost-effectiveness was 18$18,185 to make the regimen cost-effective in all patients at WTP of one time GDP per capita. The price has to be US$105 to make the regimen affordable in average patients in China.</p><p>Conclusion</p><p>Sofosbuvir/ledipasvir regimen is not cost-effective in most Chinese patients with genotype 1b HCV infection. The results vary by economic regions. Drug price of sofosbuvir/ledipasvir needs to be substantially reduced when entering the market in China to ensure the widest accessibility.</p></div

Will Sofosbuvir/Ledipasvir (Harvoni) Be Cost-Effective and Affordable for Chinese Patients Infected with Hepatitis C Virus? An Economic Analysis Using Real-World Data - Fig 2

<p><b>Probability of Cost-effectiveness in A: Treatment-Naïve and B: Treatment-Experienced patients, by economic regions in China.</b> The probability was at the willingness-to-pay of 3 GDP per capita in each region.</p

Model transition probabilities, cost inputs and utilities.

<p>Model transition probabilities, cost inputs and utilities.</p

Prices that make sofosbuvir/ledipasvir cost-effective, by four economic regions in China.

<p>Prices that make sofosbuvir/ledipasvir cost-effective, by four economic regions in China.</p