Monomeric C-reactive protein is associated with severity and prognosis of decompensated hepatitis B cirrhosis.

C-reactive protein (CRP) is an acute-phase protein produced by the liver in response to infection and during chronic inflammatory disorders. Systemic inflammation is a major driver of cirrhosis progression from the compensated to the decompensated stage. Previous studies have shown that pentameric CRP (pCRP) to be a weak predictor of disease severity and prognosis in patients with decompensated hepatitis B cirrhosis, with it being only helpful for identifying patients with a higher short-term risk of death under certain conditions. Accumulating evidence indicates that pCRP dissociates to and acts primarily as the monomeric conformation (mCRP) at inflammatory loci, suggesting that mCRP may be a potentially superior disease marker with higher specificity and relevance to pathogenesis. However, it is unknown whether mCRP and anti-mCRP autoantibodies are associated with disease severity, or progression in decompensated hepatitis B cirrhosis. In this study, we evaluated the serum levels of mCRP and anti-mCRP autoantibodies in patients with decompensated cirrhosis of hepatitis B and their association with disease severity and theoretical prognosis. The results showed that patients with high mCRP and anti-mCRP autoantibody levels had more severe liver damage and that coagulation function was worse in patients with high anti-mCRP autoantibodies. Analysis of the correlation between pCRP, mCRP and anti-mCRP autoantibody levels with Model for End-Stage Liver Disease (MELD), Albumin-Bilirubin (ALBI), and Child-Turcotte-Pugh (CTP) prognostic scores showed that mCRP was the most strongly correlated with MELD score, followed by anti-mCRP autoantibodies; conversely, pCRP was not significantly correlated with prognostic score. Therefore, mCRP and anti-mCRP autoantibodies may be more advantageous clinical indicators than pCRP for evaluating the pathological state of decompensated hepatitis B cirrhosis.

C-Reactive Protein Protects Against Acetaminophen-Induced Liver Injury by Preventing Complement Overactivation.

BACKGROUND AND AIMS: C-reactive protein (CRP) is a hepatocyte-produced marker of inflammation yet with undefined function in liver injury. We aimed to examine the role of CRP in acetaminophen-induced liver injury (AILI). METHODS: The effects of CRP in AILI were investigated using CRP knockout mice and rats combined with human CRP rescue. The mechanisms of CRP action were investigated in vitro and in mice with Fcγ receptor 2B knockout, C3 knockout, or hepatic expression of CRP mutants defective in complement interaction. The therapeutic potential of CRP was investigated by intraperitoneal administration at 2 or 6 hours post-AILI induction in wild-type mice. RESULTS: CRP knockout exacerbated AILI in mice and rats, which could be rescued by genetic knock-in, adeno-associated virus-mediated hepatic expression or direct administration of human CRP. Mechanistically, CRP does not act via its cellular receptor Fcγ receptor 2B to inhibit the early phase injury to hepatocytes induced by acetaminophen; instead, CRP acts via factor H to inhibit complement overactivation on already injured hepatocytes, thereby suppressing the late phase amplification of inflammation likely mediated by C3a-dependent actions of neutrophils. Importantly, CRP treatment effectively alleviated AILI with a significantly extended therapeutic time window than that of N-acetyl cysteine. CONCLUSION: Our results thus identify CRP as a crucial checkpoint that limits destructive activation of complement in acute liver injury, and we argue that long-term suppression of CRP expression or function might increase the susceptibility to AILI.

Association between rs2431697 T allele on 5q33.3 and systemic lupus erythematosus: case-control study and meta-analysis.

rs2431697 is located on 5q33.3, between pituitary tumor-transforming gene 1 and miR-146a. Several studies have estimated the association between rs2431697 and systemic lupus erythematosus risk. However, the results were inconsistent. A case-control study was carried out to explore the association between rs2431697 and systemic lupus erythematosus risk in a central Chinese population. Meta-analyses combining present with previous studies were conducted to further explore the association. Our case-control study included 322 cases and 353 controls. rs2431697 T allele was associated with increased risk of systemic lupus erythematosus (odds ratios (ORs) = 1.461, 95% confidence intervals (CI) 1.091-1.957, P = 0.011). The association was stronger between T allele and the risk of anti-double-stranded DNA (dsDNA)-positive systemic lupus erythematosus (OR = 2.510, 95% CI 1.545-4.077, P < 0.001). The meta-analyses included 8648 systemic lupus erythematosus patients and 10947 controls. rs2431697 T allele had an overall OR of 1.262 (95% CI 1.205-1.323, P < 0.001) under fixed-effects model. After stratified by ethnicity, I (2) reduced from 24.3 to 0 %. T allele had an OR of 1.213 (95% CI 1.145-1.284, P < 0.001) in European descendant and 1.365 (95% CI 1.259-1.480, P < 0.001) in Asian under fixed-effects model. Data on women were also extracted, and T allele had an OR of 1.337 (95% CI 1.162-1.539, P < 0.001) under random-effects model. The pooled ORs were not influenced by each study in sensitivity analyses. There were no publication biases observed in these analyses. The results from our case-control study and the meta-analyses indicate that rs2431697 T allele significantly associates with the increased risk of systemic lupus erythematosus.

Clinical relevance of plasma miR-21 in new-onset systemic lupus erythematosus patients.

BACKGROUND: Plasma miR-21 is widely investigated as biomarker in many diseases. Recent studies show that miR-21 participates in the development of systemic lupus erythematosus (SLE). The aim of this study was to evaluate the expression profile of miR-21 in the plasma of SLE patients. METHODS: Relative quantities of plasma miR-21 both in SLE patients and healthy controls were determined by relative qRT-PCR under endogenous and exogenous controls. The diagnostic value of plasma miR-21 was evaluated in SLE patients. Data of some SLE-associated clinical parameters were collected. RESULTS: Eighty participants from Central China were recruited. Forty-four participants were new-onset SLE patients and the others were healthy controls. Plasma miR-21 level in SLE patients was higher than that of healthy controls (P = 0.031). Receiver operating characteristic analysis of plasma miR-21 revealed an Area Under Curve (AUC) of 0.64 ± 0.06 (95% CI: 0.51-0.76, P = 0.03854) when differentiating SLE from healthy controls. The level of plasma miR-21 was not associated with the level of white blood cells (P = 0.4284), red blood cells (P = 0.4079), and platelets (P = 0.4961), but significantly correlated with the level of plasma complement C3 (r = -0.5297, P = 0.0004), C4 (r = -0.4732, P = 0.0020), and serum uric acid (r = 0.3932, P = 0.0121) in SLE patients. CONCLUSIONS: Plasma miR-21 in SLE patients from Central China is overexpressed. Since circulating miR-21 is aberrantly expressed in many diseases, the applying of it as a disease biomarker should be considered carefully.

Mimotope selection of blood group A antigen from a phage display 15-mer peptide library.

We select the peptide mimics of blood group A antigen by a monoclonal anti-A from a phage display 15-mer peptide library. Monoclonal anti-A was used in biopanning a phage display 15-mer peptide library. After four rounds of panning, ELISA was carried out to confirm the positive phage clones. The exogenous DNAs of the positive phages were sequenced and the corresponding amino acid sequences were deduced. Both the synthesized peptide and the phage clones were used to bind to anti-A in competitive ELISA. Erythrocyte agglutination inhibition tests were carried out to determine the mimic ability of the free synthesized peptide to the natural blood group A antigen. Computer softwares were used to simulate the interaction between the peptide and anti-A. After four rounds of biopanning, the eluted phage reached an enrichment of approximately 1600 times. Thirty-seven phage clones were chosen randomly and amplified. There were eleven clones that interacted specifically with anti-A in ELISA. DNA sequencing of the inserted oligonucleotide revealed that nine clones present a same peptide - TRWLVYFSRPYLVAT (named TRW) and each of the other two clones presented a different peptide. The synthesized free peptide TRW could inhibit the interaction of both phage displayed peptide and group A red blood cell with anti-A in competitive ELISA and hemagglutination inhibition test. Both the peptide TRW and the natural group A antigen were docked into a same cavity of anti-A in a computer simulation assay. The results indicate that peptide TRW can mimic blood group A antigen. It may be used as a proxy of natural blood group A antigen in clinical application.

Polymorphisms of the T-cell immunoglobulin and mucin domain molecule-3 are not associated with autoimmune Graves' disease in a Chinese Han Population.

OBJECTIVES: This study aimed to investigate the association between polymorphisms of T-cell immunoglobulin and mucin domain molecule-3 (TIM-3) and Graves' disease (GD) in a Chinese population. DESIGN AND METHODS: Genomic DNA was extracted from peripheral blood cells of the 182 GD patients and 150 control subjects. The TIM-3 gene polymorphic sites were genotyped. We also analyzed the relationships between the genotypes of each SNP and serum specific clinical variables. To detect whether the variants were associated with the TIM-3 expression, we further studied 40 patients by using the method of real-time quantitative reverse-transcription polymerase chain reaction (real-time RT-PCR). RESULTS: The genotype and allele frequency of each polymorphic site were not significantly different between GD and control individuals. Furthermore, it also showed no relationship between the variants and TIM-3 mRNA expression. CONCLUSIONS: Our results demonstrated that the polymorphisms of TIM-3 gene may not contribute to GD susceptibility in the Chinese Han population.

[The effect of p21 on transcription of survivin in hepatocellular carcinoma HepG2 cells and its regulation mechanism].

OBJECTIVE: To observe the inhibitory effect of cyclin-dependent kinase inhibitor p21 on regulation of survivin transcription in human liver cancer HepG cells, and explore the related mechanisms. METHODS: Doxorubicin (DOX) was used to treat HepG cells. Eukaryotic vector pEGFP-C2-p21 was transfected into HepG cells by lipofectamine and positive clones were screened out by G418. The mRNA expression of p21, p53 and survivin were detected by real-time fluorescent quantitative polymerase chain reaction (RQ-PCR). Flow cytometry was used to determine the cell cycle phases, and reverse transcription polymerase chain reaction (RT-PCR) was used to measure the levels of E2F-1 or p300. RESULTS: After treatment with DOX, the expression of p53 and p21 was increased, whereas that of survivin was reduced during 24 hours of the treatment. After transfection the p21 level was 2100.1-fold or 980.9-fold enhanced in comparison with that in HepG2 cells or HepG2-pEGFP cells. Survivin level was markedly down-regulated to 0.5% or 0.6% relative to that in the other two groups, nevertheless, significant p53 changes were not observed. Overexpression of p21 resulted in G1/G0 phase arrest (F = 31.59, P < 0.01), meanwhile, E2F-1 mRNA or p300 mRNA were less expressed compared with that in the other controls (F(E2F-1) = 125.28, P < 0.05; Fp300 = 46.01, P < 0.01). CONCLUSION: p21 could be a potential mediator of survivin suppression at transcription level in HepG2 cells, which might be through the block at G1/G0 phase and down-regulation of transcription factors E2F-1 and p300.