3) shows significant

effect of daclizumab (RR 082; CI 0

3) shows significant

effect of daclizumab (RR 0.82; CI 0.71-0.95; P = 0.007; 12 trials/cohorts) but not basiliximab (RR 0.87; CI 0.73-1.03; P = 0.11; seven trials), but this Dabrafenib mw does not seem to be a systematic effect because meta-regression does not indicate a significant effect of type of IL-2Ra (P for test of moderators = 0.67; Table 3) and inspection of the funnel plot (Supporting Fig. 4) shows that both types of studies are distributed similarly. Meta-regression also showed that concomitant use of MMF (in both arms) seems to amplify the effect of IL-2Ra (ratio of RR 0.83; CI 0.69-1.01; P = 0.06). Analysis of the type of CNI did not show significant effects, but trials in comparison 2 also had a lower RR compared to trials in comparison 1 (Table 3). This effect may be explained by the fact that MMF was used in all trials in comparison 2. After adjusting for MMF, the effect in comparison 2 alone is no longer

seen. We did not RO4929097 nmr observe significant heterogeneity in any of the analyses and observed only marginal changes of residual heterogeneity in meta-regression (Table 3). However, we observed considerable heterogeneity of overall rejection rates (i.e., the sums in the experimental and control groups of each trial) ranging from over 55%36 to less than 10%38 (see Supporting Table 2). Similar differences have also been observed in other meta-analyses concerning organ transplantation.42 We sought sources of heterogeneity using GLMM and found that the overall rejection rate was significantly higher in studies that required protocol biopsies (OR 3.10;

CI 1.93-4.99; P < 0.001; 19 trials/cohorts). These studies reported not only treated rejections but also histological signs of rejection without clinical correlates. Subgroup analysis of trials with and without protocol biopsies (five trials/cohorts with 435 patients and 14 trials with 2,526 patients, respectively) showed a significant effect on acute rejection only in studies without protocol biopsies (RR 0.81; CI 0.71-0.92) but not in studies that performed protocol biopsies (RR 0.92; CI 0.76-1.11). Meta-regression does not provide any evidence for a significant MCE公司 effect of protocol biopsies on acute rejection (P value for test of moderators 0.26) and the inspection of the funnel plot (Supporting Fig. 5) shows that the distribution of both groups of studies is comparable. Furthermore, the analysis of overall rejection rate showed that the use of MMF decreased the incidence of acute rejection in both study arms (OR 0.49; CI 0.31-0.77; P = 0.002; 19 trials/cohorts). After adjustment for use of MMF and type of biopsy the effect of IL-2Ra was still highly significant (OR 0.76; CI 0.64-0.90; P = 0.002; 19 trials/cohorts). Funnel plot analysis for acute rejection showed significant asymmetry (P = 0.01). Using the trim-and-fill method we augmented the data (Supporting Fig. 6) and these supposedly missing studies all had a risk ratio above 1.

Infants with perinatal subtype of BA have higher tissue levels of

Infants with perinatal subtype of BA have higher tissue levels of PROM1 expression than those with embryonic subtype. Conclusion: Expansion of collagen-producing PROM1pos cells within regions of

periportal fibrosis is associated with activated FGF and TGFβ pathways in both experimental and human BA. PROM1pos cells may therefore play an important role in the biliary selleck chemicals fibrosis of BA. (Hepatology 2014;60:941–953) “
“This chapter contains sections titled: Epidemiology Histopathology Etiology and pathophysiology Clinical features and diagnosis One or two diseases? Treatment of microscopic colitis References “
“The reported prevalence of cancer associated with a primary choledochal cyst ranges from 2.5%[1]to 26%.[2] Total cyst excision is the standard procedure to prevent malignant changes. An essential part of this surgery is the termination of the Trichostatin A in vivo reflux of pancreatic juice into the bile duct through the anomalous pancreaticobiliary duct union, so as to avoid activation of pancreatic enzymes involved in the pathogenesis of bile duct carcinoma. Theoretically, biliary malignancy should not develop after excision of the choledochal cyst because the presumed cause was abrogated by separation-operation. However, although the incidence of biliary malignancy after excision of choledochal cyst was 0.6% in Korean multicenter

study[3] and 0.7% in a Japanese nationwide study,[4] far lower than that associated with unresected choledochal cysts, it is still higher (about 200 times) than the incidence of biliary cancer in the general population in Japan. The main issues about the biliary malignancy after cyst excision are the perspective it might give on risk factors for oncogenesis, the relevance of cyst excision as a standard treatment and risk factors. In this issue of the Journal of Gastroenterology and Hepatology, Ohashi et al.[5] provided cumulative incidence data on subsequent biliary malignancy increase

more than 15 years after choledochal cyst excision, describe unfavorable outcomes, and provide a comprehensive review of biliary malignancy after cyst excision. However, several questions mentioned remained unanswered. The authors searched only the English literature. Because choledochal cyst is a relatively rare disease in the West and 上海皓元医药股份有限公司 more than half of the reported cases have occurred in Japan, we searched both English and Japanese language literature. Between 1970 and 2011, 58 cases were identified, and among them, data of site of malignancy can be available in 54 cases[3, 6-47] (Table 1). According to Todani’s classification, 24 of 41 patients (59%) were classified into type IVa, and 17 (42%) were into type I. The most common site of involvement in 54 cases was the hepatic duct, at or near the choledocho-enteric anastomosis (43%) followed by the intrahepatic duct (41%) and distal choledochus (17%).

To determine the potential role of immune cells in HCC developmen

To determine the potential role of immune cells in HCC development in the livers of TLR2−/− mice, the liver-infiltrating macrophages were examined by labeling these cells with F4/80 in DEN-treated WT and TLR2-deficient Fostamatinib order livers. We found that TLR2 deficiency led to a marked decrease

in the filtration of F4/80+ macrophages in the liver compared to the WT condition (Fig. 6A,B). The ASK1/p38 MAPK/NF-κB signaling pathway is a major sensor of oxidative stress that promotes apoptotic cell death.26, 27 Activation of this pathway leads to the production of cytokines that play important roles in triggering cell death and supporting senescence.28, 29 Compared to their WT littermates, TLR2−/− liver tissue showed a striking decrease in the activity of ASK1, p38 MAPK, and NF-κB (Fig. 6C,D). However, the activity of MAPK ERK1/2 was increased in TLR2−/− liver tissue. The expression of inflammatory cytokines, including IFN-γ, IL-1α, IL-1β, TNF-α, IL-6, and Cxcl-2 (a mouse ortholog of human IL-8), was markedly attenuated in TLR2−/− liver tissue

(Fig. 6E,F). These data indicate that the broad-spectrum suppression of the immune response to DEN-induced liver injury plays a critical role in the attenuated senescence and autophagy flux of TLR2−/− livers, which contributes to their enhanced susceptibility to the development of HCC. Based on the preceding observations, we suspected that restoring senescence might promote the degradation of p62 aggregates and attenuate the development of HCC

in TLR2−/− mice. Prophylactic treatment of TLR2−/− mice with IFN-γ, a typical TH1 cytokine that was recently CH5424802 identified as a positive modulator of senescence and autophagy,30-32 attenuated HCC development as indicated by a reduced number and size of tumor nodules in TLR2−/− livers (Fig. 7A,B). Indeed, IFN-γ treatment can restore senescence as indicated by an increase in the SA β-gal staining in MCE the TLR2−/− liver (Fig. 7C,D). Although IFN-γ treatment did not influence γ-H2A.X levels, it reduced the expression of PCNA and enhanced the expression of p53 and p21 in the TLR2−/− liver. Moreover, although IFN-γ treatment did not affect p16 expression, it resulted in a decreased level of pRb, a downstream inhibitory molecule of p16. Thus, IFN-γ treatment restored these two crucial senescence pathways. Moreover, the cytokine IL-1α, which can initiate and support the secretion of senescence-associated cytokines, was increased in IFN-γ-treated TLR2−/− liver tissue (Fig. 7E,F). Therapeutic administration of IFN-γ also attenuated HCC development (Fig. S2E,F) and decreased the appearance of p62-positive punctuate dots in TLR2−/− liver tissue (Fig. 8A). Indeed, the level of p62 in either the detergent-soluble or detergent-insoluble fraction of liver tissues was decreased by IFN-γ treatment (Fig. 8B,C), indicating a recovery of the suppressed autophagy flux in the TLR2−/− livers. Programmed cell deaths by either apoptosis (Fig. 8C) or autophagy (Fig.

6 months, compared with 74 months for those receiving only pacli

6 months, compared with 7.4 months for those receiving only paclitaxel, representing a 19% reduction in risk (p = .0169) with ramucirumab. Median progression-free survival was 4.4 months and 2.9 months, respectively, with a 27% reduction in risk (p < .0001). The objective response rate associated with the combination was 28% versus 16% with paclitaxel alone (p = .0001). At 6 months, the progression-free

survival rate was 36 versus 17%, and at 9 months 22 versus 10%, respectively. In addition, the disease control rate was much better with NVP-BGJ398 molecular weight ramucirumab, 80 versus 64%, respectively (p < .0001). Adverse events of grade ≥3 were somewhat greater with ramucirumab/paclitaxel, including neutropenia (40.7 vs 18.8%), leukopenia (17.4 vs 6.7%), hypertension (14.1 vs 2.4%), anemia (9.2 vs 10.3%), fatigue (7.0 vs 4.0%), abdominal pain (5.5 vs 3.3%), and asthenia (5.5 vs 3.3%). Thus, the REGARD and the RAINBOW trials clearly demonstrate that ramucirumab is an effective new option for second-line therapy of advanced GC. The epidermal growth factor receptor (EGFR) is the target of the monoclonal antibody inhibitors cetuximab and panitumumab, for the treatment of patients with metastasized

colorectal cancer without mutations of the RAS gene. Unfortunately, the addition of either cetuximab or panitumumab to standard platinum-based and fluoropyrimidine-based combination chemotherapy in unselected patients with advanced GC did Selleckchem Imatinib not provide any additional benefit to standard

chemotherapy alone and cannot be recommended for use in an unselected population with advanced esophagogastric adenocarcinoma [16, 17]. The receptor tyrosine kinase c-MET and its ligand, the hepatocyte growth factor (HGF), are involved in the regulation of multiple cellular processes including cell proliferation, invasion and angiogenesis. The HGF/c-MET signaling pathway is frequently over-expressed in GC and represents a candidate target for personalized cancer treatment. Whether or not treatment with the c-MET/HGF antibody rilotumumab in combination with a standard chemotherapy (epirubicin, cisplatin and capecitabine) significantly improves overall survival in subjects with unresectable locally advanced or metastatic MET positive gastric or gastroesophageal junction adenocarcinoma 上海皓元 is being evaluated in a phase 3, multicentre, randomized, double-blind, placebo-controlled study [18]. Recent advances in the understanding of immunology and antitumor immune responses have led to the development of new immunotherapies, including monoclonal antibodies that inhibit immune checkpoint pathways. The cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) and programmed cell death protein-1 (PD-1) pathways are two of several immune checkpoint pathways that play critical roles in controlling T-cell immune responses [19]. CTLA-4 and PD-1 are expressed by T cells.

This confirms the epidemiological evidence that variant CJD is ca

This confirms the epidemiological evidence that variant CJD is caused by BSE infection, and as such represents the only example of a human prion disease acquired from another species (Table 1). The most likely source of human exposure to BSE is the consumption

of contaminated meat products [13]. In keeping with an oral route of infection, both PrPSc and infectivity are detectable within lymphoid tissues, including the spleen, tonsil, lymph nodes, thymus and gut associated lymphoid tissue in variant CJD [9]; levels of infectivity in lymphoid tissues this website are approximately 2–3 logs lower than in brain tissue [14]. Abnormal PrP has been detected by immunohistochemistry in gut-associated lymphoid tissue within the appendix in two patients who had undergone appendicectomy up to 2 years before the onset of variant CJD [15]. These findings have lead to the suggestion that lymphocytes may carry infectivity in blood during the incubation period of variant CJD [16]. This suggestion has been reinforced by the detection of infectivity buy LY2606368 in the blood of sheep experimentally infected with BSE before the animals develop clinical signs and symptoms [17]. By February 2010,

172 cases of variant CJD have been confirmed in the UK, with 47 additional cases in 10 other countries (Table 3). The incidence of variant CJD has declined in the UK since 1999–2000; however, the number of asymptomatic infections in the UK remains uncertain; the results of earlier studies to detect abnormal PrP in tonsil and appendix tissues suggests MCE a prevalence of around 1 per 10 000 of the UK population [15,18,19]. This figure is higher than the current numbers of variant CJD cases in the UK would suggest, indicating that some variant CJD cases may have a prolonged asymptomatic carrier state, which perhaps does not result in clinical disease in all cases. Prion infectivity

in blood has been demonstrated in rodent models during the incubation period and after the clinical onset of disease [20,21]; the levels of infectivity in whole blood as measured by bioassay are around 10 ID mL−1. Most infectivity was found to be associated with white blood cells, but infectivity was also present in plasma and red blood cells [20]. Experimental BSE and scrapie infection in sheep has also demonstrated infectivity in blood during the incubation period and after the clinical onset of disease. Transmission rates of at least 40% were reported following blood transfusion, with most transmissions occurring from blood samples taken during the second half of the incubation period; transmission of scrapie was also reported with a relatively small volume of a buffy coat preparation [17]. The Transfusion Medicine Epidemiology Review was established in the UK to investigate the possibility of transmission of variant CJD by blood transfusion [22].

16 Indeed, we found that IA had a longer half-life than IFNα and

16 Indeed, we found that IA had a longer half-life than IFNα and exhibited hepatic tropism. Liver targeting was ascertained by pharmacokinetic and biodistribution studies and also by the more intense activation of hepatic ISGs when using pIA for liver transduction than when employing pIFN or pALF, despite the fact that albumin-IFNα has a longer half-life than IA. Unexpectedly, fusion of IFNα to

ApoA-I markedly changed check details the biological properties of IFNα. Thus, experiments in murine fibroblasts showed that the cytotoxic effects of either HDL-IA or rIA were significantly lower than those of IFNα. Moreover, although IFNα induces activation-dependent cell death in T lymphocytes,19, 20 this effect is much lower SB431542 mouse with IA. Thus, IA facilitates lymphocyte proliferation in response to mitogens. These in vitro results are in accordance with data from in vivo killing assays where we found that the adjuvant efficacy of IA was far superior to that of IFNα. These findings indicate that the immunostimulatory properties of IA are not merely related to its persistence in circulation but rather to the fact that IA does not induce cell death in activated lymphocytes as does IFNα. In order to explore whether the interaction of IA with SR-BI

(the main ApoA-I receptor11) is required for its enhanced adjuvant activity, we performed in vivo killing assays after LacZ vaccination using as adjuvants pIA or pIFN in SR-BI+/−, SR-BI−/−, and wildtype mice. Our data showing that the adjuvant effect of IA was superior in wildtype animals but not in SR-BI−/− animals indicates that ligation of IA to SR-BI is needed for the fusion protein to display its potent immunostimulatory properties. The lower cytotoxic activity of IA compared to IFNα is also reflected by their different influence on hematopoiesis. One of the side effects of IFNα treatment is the development of leukopenia and thrombocytopenia, which limits its use in patients who already have diminished blood counts.8 In contrast to 上海皓元医药股份有限公司 IFNα, the administration of IA did not significantly reduce the number of platelets and only

caused a transient drop in leukocytes, which rapidly recovered. Moreover, the bone marrow progenitor compartment was induced to proliferate in the absence of significant modifications in the number of circulating leukocytes and platelets, suggesting that IA might stimulate myelo and thrombopoiesis. The safety of IA was also demonstrated in mice whose liver was transduced with an AAV vector encoding IA. No hematological or biochemical toxicity was found in these animals at day 30 after vector administration. The improvement of the pharmacological profile of IFNα resulting from its linkage to ApoA-I was not reproduced by fusion to other apolipoproteins present in HDLs. Anchoring IFNα to ApoF generated an unstable molecule which was not expressed.

Muskin, MD[1] The neurologist is well known to those who work in

Muskin, MD.[1] The neurologist is well known to those who work in the field of headache http://www.selleckchem.com/products/epz015666.html and so is one of the contributing authors, Robert G. Kaniecki, MD. Drs. Green and Kaniekci are responsible for 2 of the 3 headache chapters in the book Migraine and Tension-Type Headache. The third chapter, Chronic Daily Headache, is written by

Robert P. Cowan, MD, a neurologist from Stanford University. The 3 headache chapters cover most of the headache types commonly seen in practice. Stress is an important circumstance contributing to headache onset[2] as well as a trigger of individual headaches[3] and 2 chapters in the book are devoted to it, one on Stress and Headache and the other on Stress Management. The latter chapter covers extensively the stress-management techniques of relaxation therapy, biofeedback, cognitive behavior therapy, and coping skills. The introduction to the chapter on Working With Personality and Personality Disorders in the Headache Patient contains the contentious statement: “Headache patients

BGJ398 cell line in particular demonstrate excessive personality dysfunction, the headache often manifesting the patient’s interpersonal stress.” The chapter is written by a psychiatrist, and the statement caught my attention because I was confronted with a similar notion when I was a resident in psychiatry as part of my neurology training and never clearly understood its meaning. There is also a 上海皓元医药股份有限公司 chapter in the book that is only marginally related to the main topic of the book, dealing with Complementary and Alternative Medicine (CAM) Approaches to Headache. It covers lifestyle, exercise, and dietary considerations, body-centered, mind-centered, and mind/body-centered approaches, alternative medical systems, homeopathy, and manual therapies. The remaining chapters have a more traditional psychiatric content and deal with

mood disorders, anxiety disorders, somatoform disorders, psychosis, and, last but not least, substance dependence or addiction. The latter chapter also contains an interesting section on malingering. The book reminds me of the one on Psychiatric Aspects of Headache, edited by Charles S. Adler, Sheila M. Adler, and Russell C. Packard, published in 1987 to which I contributed a chapter on The Physiology and Biochemistry of Stress in Relation to Headache.[4] The latter book is different in the sense that the chapters are mostly written by specialists working in headache and does not cover the psychiatric aspects of headache as extensively as the present book. The book, as edited by Green and Muskin, is very readable and full of information relevant to practice. Regrettably, its structure, in the sense of a logical build up of the chapters, leaves somewhat to be desired. Nevertheless, I highly recommend the book to anybody interested in headache whether working in general medical or dental practice, neurological, psychiatric, or psychological practice, or in specialty headache practice.

The difference in biological half-life varies a lot

The difference in biological half-life varies a lot selleck inhibitor among individuals [6]. The levels of FVIII between two subjects may differ dramatically, for example, 48 h post infusion and time for the clotting factor level to decline to 1% may differ by more than 2 days. Consequently, dose and dosing during prophylaxis should

be individualised and based on individual PKs. Prophylaxis performed without ‘PK-thinking’ and implementation is therefore not recommended if treatment is meant to become optimised. This is also very true for factor replacement during surgery, irrespective of whether it is dosed as intermittent injections or as continuous infusion. In the largest prophylaxis study ever published (the recent comparison between The Netherlands

and Sweden), one main conclusion was that prophylaxis should be tailored individually and has a potential to save money at sustained efficacy [18]. The evidence in favour of PK parameters as a good surrogate for clinical efficacy and for the best use of money is thus overwhelming and was also convincingly shown when going from so-called standard dosing (every second day) to daily dosing [19]. A new interesting but also challenging era in haemophilia treatment is just around the corner. Long-acting FVIII and IX products will be available for treatment during the coming years. PK of these products differ substantially from traditional products in that the former, especially FIX where half-life is prolonged by around BMS-907351 in vivo five times, display a long tail-off period during which levels are quite low for many hours provided that dose intervals are longer than with traditional products. The risk for breakthrough bleeds is obvious, especially if the patient is performing vigorous physical activity. Given the discussion above, the use of PK has become obligatory to control factor levels during the days post infusion and

to monitor the risk of bleeding. MCE Another way to dose long-acting products is to keep the standard interval and dose, and instead, increase the trough level. This will, in a way, give the possibility to cure haemophilia. The use of PK calculations in routine clinical practice is jeopardised by the need for prolonged and frequent sampling to obtain fully reliable PK curves. However, this hurdle has been overcome by introducing population PK where only a few samples are needed [20, 21]. Introduction of convenient IT solutions (Apps) will certainly facilitate a more general use of PK at haemophilia centres. PK parameters are good surrogates for clinical efficacy and therefore PK should be used in haemophilia when dosing is determined. This is the only way to introduce evidence-based prophylaxis and to use this very costly therapy in the optimal way. PKs of FVIII and FIX are age dependent and individual, which also underlines the importance. Prophylactic treatment of haemophilia aims to prevent bleeding and maintain normal joint status [22].

Clinical outcomes, defined as a 2 point CTP progression, variceal

Clinical outcomes, defined as a 2 point CTP progression, variceal bleeding, ascites, hepatic encephalopathy, or liver-related death, occurred in 54 patients. Results. Patients ranged from DSI 9 (normal) to 40 (severe dysfunction). ROC curves showed that DSI could identify

patients with medium/large varices (c-statistic 0.82), and could predict which patients would have clinical outcomes (c-statistic 0.83), and DSI >25 was the optimum cutoff for both. DSI >25 had a higher balanced accuracy than cirrhosis by biopsy (Ishak F5F6), and the PPV for identifying medium/large varices increased 41% relative to Romidepsin research buy biopsy and the PPV for predicting outcomes increased 47% (Table 1). Conclusions. A dual cholate liver function test yielding a DSI could outperform histologic fibrosis stage in identifying patients with medium/large varices and in predicting clinical outcomes in chronic HCV patients. Identifying Med/Lg Varices Sens. Spec. PPV NPV Balanced Accuracy Biopsy (Ishak F5-F6) 77% 60% 18% 96% 69% DSI>25 77% 74% 25% 97% 76% Predicting Outcomes Sens. Spec. PPV NPV Balanced Accuracy Biopsy (Ishak F5-F6) 72% 66% 41% 88% 69% DSI>25 74% 84% 60% 91% 79% Disclosures: Steve M. Helmke – Patent Held/Filed: University of Colorado Gregory T. Everson – Advisory

Committees or Review Panels: Roche/Genentech, Merck, HepC Connection, Roche/Genentech, Selleckchem AZD6244 Merck, HepC Connection; Board Membership: HepQuant LLC, PSC Partners, HepQuant LLC, PSC Partners; Consulting: Roche/Genentech, BMS, Gilead, Roche/Genentech, Bristol-Myers Sguibb, Abbott; Grant/Research Support: Roche/Genentech, Pharmassett, Vertex, GSK, Schering-Plough, Bristol-Myers Sguibb, Tibotec, GlobeImmune, Pfizer, Abbott, Conatus, Zymogenetics, PSC Partners, Roche/Genentech, Pharmassett, Vertex, GSK, medchemexpress Schering-Plough, Tibotec, Globeimmune, Pfizer, Gilead, Conatus, Zymogenetics, PSC Partners,

Abbott; Management Position: HepQuant LLC, HepQuant LLC; Patent Held/Filed: Univ of Colorado, Univ of Colorado The following people have nothing to disclose: Jennifer DeSanto, Andrea Herman, Asmeen Bhatt, Shannon Lauriski Purpose- This study was undertaken to evaluate short-term survival as well as mid-term outcome after TIPS in patients who successfully underwent the procedure for variceal bleeding. Materials and Methods- Thirty nine consecutive patients with liver cirrhosis who underwent TIPS creation for treatment of vasoactive drug and endoscopy refractory variceal hemorrhage within 24 hours of acute variceal bleed were identified among all patients undergoing TIPS (N=87) over a two year period at our institute. Technical success was defined as successful creation of a shunt between the hepatic vein and an intrahepatic portal venous branch. Hemodynamic success was defined as reduction in the portosystemic pressure gradient to an absolute value less than 12 mm Hg.

Increased induction of

STATs and IRF9 was also observed a

Increased induction of

STATs and IRF9 was also observed after IFN-α treatment in Pol-expressing Huh7 cells but was much weaker than that observed in control cells (Fig. 2B). The levels of STAT1 Ser727 phosphorylation were clearly repressed by transfection of Huh7 cells with Pol; however, tyrosine phosphorylation of STAT1/2 was not affected. To further investigate the effect of Pol on the tyrosine phosphorylation-induced STAT1-STAT2 heterodimerization, we performed co-immunoprecipitation (co-IP) experiments in Huh7 cells transfected with increasing amounts of Pol (Fig. 2C) and in Dox-regulated HepAD38 cells (Fig. 2D). The results showed that STAT1-STAT2 interaction selleck chemicals llc in response to IFN-α was consistently observed in cells with or without Pol. Meanwhile, Flag-Pol was not detected in the immune complexes precipitated with anti-STAT1

or Ruxolitinib mw anti-STAT2 Abs, indicating no direct interaction between Pol and activated STAT1/2. Moreover, there was not much difference in IFN-α–induced heterodimer formation between cells expressing Pol and control cells (Fig. 2E,F), indicating that Pol does not affect the IFN-α–stimulated STAT1-STAT2 heterodimerization. IFN-α-induced phosphorylation of the serine residue at position 727 (Ser727) of STATs contributes critically to their transcriptional activity.12, 13 Although still controversial, PKC-δ, p38 and ERK have been reported to function as kinases that regulate Ser727 phosphorylation.14, 15 To elucidate the mechanism by which Pol interferes with STAT1 Ser727 phosphorylation, we examined the effect of Pol on IFN-α–induced phosphorylation of PKC-δ, p38 and ERK (Fig 3A). The results showed that Pol only inhibited PKC-δ but not p38 or ERK phosphorylation in IFN-α–stimulated Huh7 cells. Rottlerin, a selective inhibitor of PKC-δ, was used to verify the role of PKC-δ in Ser727 phosphorylation of STATs (Fig. 3B), and the 上海皓元医药股份有限公司 data demonstrate that PKC-δ is specifically required

for the Ser727 phosphorylation, but not for STAT tyrosine phosphorylation. IFN-α–stimulated PKC-δ phosphorylation was also found to be impaired in HepG2.215 cells compared with that in HepG2 cells (Fig 3C), but was restored by Pol siRNA transfection (Supporting Fig. 5A). In addition, we investigated whether Pol inhibits IFN-α signaling by regulating the level of STAT3, as it was reported to be a negative regulator of the type I IFN response.16 Little difference in the basal expression level and IFN-α–induced tyrosine phosphorylation of STAT3 was observed between the cells with or without Pol; however, PKC-δ–dependent Ser727 phosphorylation of STAT3 was inhibited by Pol in a dose-dependent manner (Fig. 3D). Furthermore, less STAT1 was coprecipitated with PKC-δ from lysates of Pol-expressing IFN-α–treated cells (Fig. 3E), and Pol was found to interact with the catalytic domain of PKC-δ (Fig. 3F and Supporting Fig. 5B).