Evaluation of Serum Alpha-Fetoprotein Level in Chronic Hepatitis C Patients.

BACKGROUND: Hepatitis C virus (HCV) infection represents an important risk factor for hepatocellular carcinoma (HCC). Alpha-fetoprotein (AFP) is a widely used biomarker for HCC. However, elevated serum AFP levels in different statuses of chronic hepatitis C (CHC) is not well defined. This study aimed to evaluate the relationship between AFP levels and HCV viral load in CHC patients. We also analyzed the correlation between three liver func-tion enzyme levels (AST, ALT, GGT) and HCV RNA viral load in CHC patients. METHODS: A total of 279 patients infected with HCV were included in this study. Patients were divided into two groups: HCV RNA positive and HCV RNA negative group. Serum HCV RNA load was measured by Quantitative Real-time PCR. Electrochemiluminescence assay (ECLA) was used to determine the serum AFP levels. The differences between two groups in AFP levels and biochemical profile (AST, ALT, GGT) was evaluated. RESULTS: The HCV RNA-positive group had significantly higher serum AFP levels than the negative groups (12.61 vs. 4.72 ng/mL, p < 0.0001). There was no correlation between AFP levels and HCV RNA viral load in HCV infection patients (p = 0.92). A significant correlation was observed between serum ALT (r = 0.243, p = 0.005), GGT (r = 0.212, p = 0.016) levels and HCV RNA viral load. Poor correlation (r = 0.148, p = 0.093) was found between AST levels and HCV RNA viral load. Interestingly, there was a positive correlation (r = 0.337, p < 0.001) between ALT and AFP levels in the whole study population. CONCLUSIONS: We concluded that serum HCV RNA positive patients were candidates for therapeutic prevention of HCC and liver inflammation regardless of the HCV RNA viral load. Furthermore, higher burden of HCV viral load was associated with more severe liver damage.

Relationship between serum quantitative HBsAg and HBV DNA levels in chronic hepatitis B patients.

Serum hepatitis B surface antigen (HBsAg) level has been developed as an important marker to predict treatment outcome recent years. The authors aimed to identify the correlation between quantitative HBsAg and hepatitis B virus (HBV) DNA level in chronic hepatitis B (CHB) patients and explore whether quantitative HBsAg can be used as a surrogate marker of serum HBV DNA for CHB patients. One hundred seventy-three patients were included in this study. Patients were divided into two groups: Hepatitis B e antigen (HBeAg) positive and negative patients. There was a positive correlation between quantitative HBsAg and HBV DNA level in HBeAg positive patients (r = 0.509, P < 0.001) and poor correlation in HBeAg negative patients (r = 0.176, P = 0.096). Interestingly, completely no correlation (r = -0.01, P = 0.994) was found in younger HBeAg negative patients (<40 years old), whereas in older HBeAg negative patients (>40 years old) there is a positive correlation (r = 0.448, P = 0.003). Mean HBsAg titer and Alanine aminotransferase (ALT) level were significantly higher in HBeAg positive group (3.81 log10 IU/mL; 105 IU/mL) than in negative group (2.85 log10  IU/mL; 32 IU/mL) (P <  0.001). We concluded that quantitative HBsAg could reflect HBV DNA level in HBeAg positive patients, but could not surrogate for HBV DNA level in HBeAg negative patients. Our study improves understanding of the relationship between HBsAg titers and HBV DNA levels in CHB patient and may have implications for future treatment algorithms evaluating the HBsAg titers in both HBeAg positive and negative patients.

Ultrasensitive detection of serum hepatitis B virus by coupling ultrafiltration DNA extraction with real-time PCR.

BACKGROUND: A simple and reliable DNA extraction of hepatitis B virus (HBV) is critical in developing an ultrasensitive detection method for HBV infection. Current commercially available serum Hepatitis B Virus (HBV) DNA extraction methods are time-consuming, expensive and/or require specialized equipment, which hinders wide adoption of clinical laboratories. This study offers a report on an ultrasensitive HBV DNA detection method by coupling serum HBV DNA extraction by ultrafiltration (UF) with real-time PCR (qPCR) detection. METHODS: Serum proteins were precipitated by phenol to release HBV DNA in the supernatant which was then transferred to the UF devices. The resultant DNA concentrate was eluted and released into qPCR pre-mixture. The UF-qPCR assay performance, including recovery rate, linearity, detection sensitivity, precision and diagnostic accuracy that compared to the CAP-CTM V2.0 assay by analyzing batched low viral load clinical samples was evaluated. RESULTS: The recovery rate of the UF-based HBV DNA extraction method was above 80%. The assay linearity was demonstrated with a slope of 0.95 and R2 values of 0.99. Limit-of-detection (LOD) of the UF-qPCR assay was determined to be 12.1IU/ml. The coefficient of variation (CV) of HBV quantitation for high, low and limit titer samples was 2.28%, 5.77% and 25.59%, respectively. Accuracy of the UF-qPCR assay was confirmed with the reference panel, and there was a strong correlation between these two methods (R2 = 0.55, p < 0.01). CONCLUSIONS: The UF-qPCR assay is reliable, highly sensitive, affordable and time-saving, and the method can be used for ultrasensitive detection of serum HBV.

Inhibition of Sonic Hedgehog Signaling Pathway by Thiazole Antibiotic Thiostrepton Attenuates the CD44+/CD24-Stem-Like Population and Sphere-Forming Capacity in Triple-Negative Breast Cancer.

BACKGROUND/AIM: Triple-negative breast cancer (TNBC) represents a particular clinical challenge because these cancers do not respond to endocrine therapy or other available targeted agents. The lack of effective agents and obvious targets are major challenges in treating TNBC. In this study we explored the cytostatic effect of thiazole ring containing antibiotic drug thiostrepton on TNBC cell lines and investigated the molecular mechanism. METHODS: Cell viability was measured by MTT assay. Cell surface marker was monitored by FCM. Western blot was applied to assess the protein expression levels of target genes. RESULTS: We found that thiostrepton remarkably suppressed the CD44+/CD24- stem-like population and sphere forming capacity of TNBC cell lines. Notably, we showed for the first time that thiostrepton exerted its pharmacological action by targeting sonic hedgehog (SHH) signaling pathway. Thiostrepton repressed SHH ligand expression and reduced Gli-1 nuclear localization in TNBC cell line. Furthermore, the downstream target of SHH signaling undergone dose-dependent, rapid, and sustained loss of mRNA transcript level after thiostrepton treatment. Finally, we showed that SHH ligand was essential for maintaining CD44+/CD24- stem-like population in TNBC cell line. CONCLUSION: We conclude that thiostrepton suppresses the CD44+/CD24- stem-like population through inhibition of SHH signaling pathway. Our results give a new insight into the mechanism of thiostrepton anti-tumor activity and suggest thiostrepton as a promising agent that targets hedgehog signaling pathway in TNBC.

[Association of L-selectin gene polymorphism with susceptibility to coronary heart disease].

OBJECTIVE: To investigate the possible association between L-selectin gene P213S polymorphism and coronary heart disease (CHD) in Chinese population. METHODS: In total 212 CHD patients diagnosed by angiography and 230 healthy controls were studied. The genotype and allele frequencies of L-selectin gene polymorphism were assayed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: The frequency of the L-selectin gene 213P allele in CHD patients was significantly higher than that in the control group (77.59% vs 69.35%, P=0.006). Compared with the SS genotype, PP homozygote had a significantly increased CHD risk (OR=2.70 and OR=2.15 using unadjusted and adjusted Logistic regression models, respectively). No association was found between the severity of CHD and the Lselectin gene P213S polymorphism CONCLUSION: Our findings suggest that L-selectin gene 213P mutant allele might contribute to susceptibility of Chinese individuals to contract CHD.

[Association of C-159T polymorphism in promoter region of CD14 and coronary heart disease].

OBJECTIVE: To investigate the distribution of CD14 promoter gene -159(C>T) polymorphism in Hubei Han population of China and analyze the association of CD14 polymorphisms with coronary heart disease (CHD). METHODS: Genotypes of CD14 were typed in 162 CHD patients and 196 controls by polymerase chain reaction-restriction fragment length polymorphism. Selected coronary angiography was performed in 162 CHD patients. RESULTS: CD14 promoter -159 genotype frequencies of CC, CT and TT were 27.4%, 45.6%, 27.0% and 14.8%, 46.5%, 38.7% in normal control group and CHD group respectively. Genotype distribution was in accordance with Hardy-Weinberg equilibrium. There existed statistically significant difference in frequencies of allele and genotype in CD14 C-159T polymorphism between CHD group and control group (Genotype: Chi2=0.654, P < 0.05, CT vs CC, OR=1.245, 95%CI: 1.001-1.473, TT vs CC, OR=2.374, 95%CI 2.012-2.649; Allele: Chi2=0.547, P < 0.05, T vs C, Chi2=0.547, P < 0.05, OR=3.105, 95%CI: 2.493-3.539). The distributions of allele and genotype in CD14 -159(C>T) were of statistically significant difference between non-myocardial infarction subgroup and myocardial infarction subgroup (Genotype: Chi2=0.782, P < 0.05, CT vs CC, OR=2.375, 95%CI: 2.017-2.689, TT vs CC, OR=3.459, 95%CI: 3.003-3.846. Allele: Chi2=2.374, P < 0.05, T vs C, Chi2=2.374, P < 0.05, OR=4.011, 95%CI: 3.814-4.279). However, no statistically significant difference was found among the subgroups of oneìtwo and three stenosed vessels. CONCLUSION: The T allele of the C-159T polymorphism of CD14 gene may be a risk factor for myocardial infarction.