ABSTRACT
PURPOSE: To analyze the correlation of polymorphisms of toll-like receptor 7 (TLR7) (rs179009) and toll-like receptor 9 (TLR9) (rs187084) in hepatitis C virus (HCV) infections in the Han population. MATERIALS AND METHODS: The genotypes of TLR7IVS2-151 in HCV infection were detected by Sanger sequencing using polymerase chain reaction-restriction fragment length polymorphism to determine the TLR9 T-1486C single nucleotide polymorphisms (SNP) for all enrolled patients. RESULTS: We found no significant difference between males with spontaneous clearance of HCV versus those chronically infected [chi2=2.71, p=0.10, odd ratios (OR)=0.58, 95% confidence interval (CI) 0.31-1.11]. However, significant differences were found for the distribution of TLR7 (rs179009) in females (chi2=9.46, p=0.01). In females, a significant difference was also found between chronic hepatitis C and those with spontaneous clearance of HCV in terms of TLR7 IVS2-151G/A allele frequencies (chi2=9.50, p=0.00, OR=0.46, 95% CI 0.28-0.75). In HCV-infected patients, no significant association was found between the frequency of TLR9 genotypes and alleles. CONCLUSION: The site of TLR7 IVS2-151 (rs179009) G/A may be a factor for susceptibility of chronic HCV in the female Han population. TLR9T-1486C (rs18084) SNP may not play a major role in HCV infection. However, individual risk profiles for HCV infection did vary by sex and this relationship should be further investigated.
Subject(s)
Female , Humans , Male , Alleles , China , Confidence Intervals , Gene Frequency , Genotype , Hepacivirus , Hepatitis C , Hepatitis C, Chronic , Hepatitis , Methods , Polymorphism, Single Nucleotide , Toll-Like Receptor 7 , Toll-Like Receptor 9 , Toll-Like ReceptorsABSTRACT
The effects of tacrolimus postconditioning on protein-serine-threonine kinases (Akt) phosphorylation and apoptotic cell death in rats after spinal cord ischemia-reperfusion injury were investigated. Ninety male SD rats were randomly divided into sham operation group, ischemia-reperfusion group and tacrolimus postconditioning group. The model of spinal cord ischemia was established by means of catheterization through femoral artery and balloon dilatation. The spinal cord was reperfused 20 min after ischemia via removing saline out of balloon. The corresponding spinal cord segments were excised and determined for Akt activity in spinal cord tissue by using Western blotting at 5, 15, and 60 min after reperfusion respectively. Spinal cord tissue sections were stained immunohistochemically for detection of the phosphorylated Akt expression at 15 min after reperfusion. Flow cytometry was applied to assess apoptosis of neural cells, and dry-wet weights method was employed to measure water content in spinal cord tissue at 24 h after reperfusion. The results showed that the activities of Akt in tarcolimus postconditioning group were significantly higher than those in ischemia-reperfusion group at 5, 15, and 60 min after reperfusion (P<0.05, P<0.01). The Akt activities reached the peak at 15 min after reperfusion in ischemia-reperfusion group and tacrolimus postconditioning group. The percentage of apoptotic cells and water content in spinal cord tissue were significantly reduced (P<0.01) in tacrolimus postconditioning group as compared with those in ischemia-reperfusion group at 24 h after reperfusion. It is concluded that tacrolimus post-conditioning can increase Akt activity in spinal cord tissue of rats, inhibit apoptosis of neural cells as well as tissue edema, and thereby alleviate spinal cord ischemia-reperfusion injury.
Subject(s)
Animals , Male , Rats , Apoptosis , Immunosuppressive Agents , Pharmacology , Therapeutic Uses , Phosphorylation , Protein Serine-Threonine Kinases , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Rats, Sprague-Dawley , Reperfusion Injury , Drug Therapy , Metabolism , Spinal Cord , Metabolism , Pathology , Spinal Cord Ischemia , Drug Therapy , Metabolism , Tacrolimus , Pharmacology , Therapeutic Uses , Up-RegulationABSTRACT
<p><b>OBJECTIVE</b>To investigate the therapeutic efficiency of antiviral treatment with pegylated-interferon (Peg-IFN) for hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) and to explore whether liver histopathological features or other factors influence the HBeAg seroconversion treatment response.</p><p><b>METHODS</b>Eighty HBeAg-positive CHB patients with diagnosis confirmed by liver puncture were treated with Peg-IFN(2a or 2b)body weight dose, once weekly). At treatment week 48, the rate of HBeAg seroconversion was determined and used to analyze the influence of liver histopathological features (liver biopsy assessment of: inflammation, graded G0 to G4; fibrosis stage, graded S0 to S4), sex, age, differential levels (pre-treatment baseline vs. week 48 post-treatment) of serum alanine transferase (ALT), and HBV DNA, by binary logistic analysis.</p><p><b>RESULTS</b>At week 48, the overall rate of HBeAg seroconversion was 30.0%. The rate of HBeAg seroconversion gradually advanced with increased liver inflammation (X2 = 8.435, P = 0.015): 9.09% of the 22 patients with G1; 31.58% of the 38 patients with G2; 47.30% of the 19 patients with G3; the one patient with G4. In contrast, the rate of HBeAg seroconversion showed a much weaker association with liver fibrosis (X2 = 5.917, P = 0.116). Only baseline HBeAg level, and no other baseline index, was significantly different between the patients who achieved HBeAg seroconversion and those who did not. Liver inflammation and baseline HBeAg level were identified as influencing factors of HbeAg seroconversion in response to Peg-IFN treatment.</p><p><b>CONCLUSION</b>Peg-IFN therapy induces a higher rate of HBeAg seroconversion in HBeAg-positive CHB patients with severe liver inflammation; histological analysis of pre-treatment liver biopsies may help to identify patients most likely to benefit from the antiviral regimen.</p>
Subject(s)
Adult , Female , Humans , Male , Antiviral Agents , Therapeutic Uses , Hepatitis B e Antigens , Blood , Hepatitis B, Chronic , Blood , Drug Therapy , Pathology , Interferon-alpha , Therapeutic Uses , Liver , Pathology , Polyethylene Glycols , Therapeutic Uses , Recombinant Proteins , Therapeutic Uses , Serologic TestsABSTRACT
The effects of tacrolimus postconditioning on protein-serine-threonine kinases (Akt) phosphorylation and apoptotic cell death in rats after spinal cord ischemia-reperfusion injury were investigated. Ninety male SD rats were randomly divided into sham operation group, ischemia-reperfusion group and tacrolimus postconditioning group. The model of spinal cord ischemia was established by means of catheterization through femoral artery and balloon dilatation. The spinal cord was reperfused 20 min after ischemia via removing saline out of balloon. The corresponding spinal cord segments were excised and determined for Akt activity in spinal cord tissue by using Western blotting at 5, 15, and 60 min after reperfusion respectively. Spinal cord tissue sections were stained immunohistochemically for detection of the phosphorylated Akt expression at 15 min after reperfusion. Flow cytometry was applied to assess apoptosis of neural cells, and dry-wet weights method was employed to measure water content in spinal cord tissue at 24 h after reperfusion. The results showed that the activities of Akt in tarcolimus postconditioning group were significantly higher than those in ischemia-reperfusion group at 5, 15, and 60 min after reperfusion (P<0.05, P<0.01). The Akt activities reached the peak at 15 min after reperfusion in ischemia-reperfusion group and tacrolimus postconditioning group. The percentage of apoptotic cells and water content in spinal cord tissue were significantly reduced (P<0.01) in tacrolimus postconditioning group as compared with those in ischemia-reperfusion group at 24 h after reperfusion. It is concluded that tacrolimus post-conditioning can increase Akt activity in spinal cord tissue of rats, inhibit apoptosis of neural cells as well as tissue edema, and thereby alleviate spinal cord ischemia-reperfusion injury.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of core protein of hepatitis C virus (HCV) on the expression of hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF).</p><p><b>METHODS</b>Huh7.5.1 cells were transfected with plasmid flag2B-core carrying HCV core gene, expression of HIF-1alpha and VEGF were measured by reverse transcription-polymorphism chain reaction (RT-PCR) and western blot. Enzyme link immunoabsorbent assay (ELISA) were used to detect the level of VEGF in the supernatants.</p><p><b>RESULTS</b>The expression of HIF-1alpha and VEGF mRNA and protein were upregulated after flag2B-core was transfected into Huh7.5.1 cells, and VEGF level in the supernatant was significant elevated as compared to controls [(654.5+/-43.7) pg/ml vs (365.9+/-26.8) pg/ml, t = 653.1%, P less than 0.01]. The expression of HIF-1alpha and VEGF mRNA and protein were downregulated after flag2B-core and HIF-1alpha siRNA were co-transfected into Huh7.5.1 cells, and VEGF level in the supernatant was significantly reduced as compared to controls [(389.2+/-29.6) pg/ml vs (768.8+/-47.3)pg/ml, t = 1330.22, P less than 0.01].</p><p><b>CONCLUSIONS</b>HCV core protein enhances the expression of HIF-1alpha and VEGF. HCV may regulate the expression of HIF-1alpha and VEGF via the core protein.</p>
Subject(s)
Humans , Cell Line, Tumor , Hypoxia-Inducible Factor 1, alpha Subunit , Metabolism , RNA, Small Interfering , Transfection , Vascular Endothelial Growth Factor A , Metabolism , Viral Core Proteins , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of tacrolimus on expression of heat shock protein 70 (HSP 70) after spinal cord injuries (SCI) in rats and the relationship between expression of HSP 70 and apoptosis of neural cells.</p><p><b>METHODS</b>Seventy-two male rats were divided randomly into three groups: the sham-operation group, the injury group and the group treated with tacrolimus, and the latter two groups were SCI with a weight-drop impactor at the T(10) vertebrae level (10 g weight was dropped from a 4.0 cm height). The tacrolimus group was injected with tacrolimus 5 minutes after SCI, while the other groups received 0.9% saline likewise. The inclined plate and BBB (Basso, Beattie and Bresnahan) scales were used to evaluate hindlimb neurological function. The expression of HSP 70 mRNA after SCI was detected by using reverse transcription polymerase chain reactions (RT-PCR) and immunohistochemistry staining was performed to determine the protein expression of HSP 70 and Caspase-3. The apoptosis of neural cells was assessed with the terminal deoxynucleotidyl transferase-mediated deoxyuredine triphosphate-digoxin nick end labeling (TUNEL) method.</p><p><b>RESULTS</b>Compared with the injury group, the expression of HSP 70 was significantly higher in the tacrolimus group, and the peak expression of HSP 70 mRNA and protein was respectively observed at 6, 24 h after SCI. Caspase-3-positive or TUNEL-positive cells were significantly less in the tacrolimus group than in the injury group. Neurological function score of the tacrolimus group was significantly better than that of the injury group.</p><p><b>CONCLUSIONS</b>Tacrolimus may inhibit activity of Caspase-3, attenuate apoptosis of neural cells and ameliorate neurological function recovery after SCI by inducing high expression of HSP 70.</p>