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.

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.

[Development of a GeXP based multiplex RT-PCR assay for simultaneous differentiation of nine human hand food mouth disease pathogens].

A multiplex RT-PCR assay based on GeXP system was developed in order to detect simultaneously human enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) and other coxsackieviruses (CVA4, 5, 9 and 10, CVB1, 3 and 5). Enterovirus detection was performed with a mixture of 12 pairs of oligonucleotide primers including one pair of published primers for amplifying all known pan-enterovirus genomes and eleven primer pairs specific for detection of the VP1 genes of EV71, C A16, CVA4, CVA5, CVA9, CVA10, CVB1, CVB3 and CVB5, respectively. The specificity of multiplex RT-PCR system was examined using enterovirus cell cultures and positive strains identified previously from hand-foot-and-mouth disease (HFMD) patients. Serial dilution of titrated EV71 and C A16 cell cultures and in vitro transcripted RNA of enterovirus VP1 regions were used to detect the sensitivity of the multiplex RT-PCR system. The limit of detection for this multiplex RT-PCR system was 10(0.5) TCID50/microL for EV71 and C A16 cell cultures and 1000 copies for in vitro transcripted RNA of nine viruses per assay. This multiplex RT-PCR assay is a rapid, sensitive and specific assay for the diagnosis of common enterovirus infection in cases of HFMD outbreak and is also potentially useful for molecular epidemiological investigation.

[Studies on the effect of Ginkgo biloba extracts on NF-kappaB pathway].

OBJECTIVE: To investigate NF-kappaB and IkappaBalpha activities in HL-60 induced by TNF-alpha in order to understand the molecular mechanism of GbE in asthma treatment. METHODS: The amount of IkappaBalpha in HL-60 cells stimulated by TNF-alpha and GbE was measured by western blotting. Plasmid pNF-kappaB-LuC was transfected and NF-kappaB activity was analyzed by measuring the expression level of luciferase. RESULTS: It showed in the luciferase assay that the activity of NF-kappaB could significantly be suppressed in HL-60 cells after the pretreatment with CGbE. However, the phosphorylation and subsequent degradation of IKBalpha induced by TNF-alpha can not be inhibited in HL-60 cells even we prolonged the treatment time or increased the concentration of GhE. CONCLUSION: GhE can suppress the NF-kappaB gene expression actively on independent of NIK/ IKK/ IkappaBalpha pathway in HL-60 cells.

[Variances of leptin mRNA in the adipose tissue of NAFLD rats intervened with the extracts of Polygonum cuspidatum compound].

The NAFLD rats were intervened with the extracts of Polygonum cuspidatum compound for 4 weeks. The reverse transcription and real-time quantitative polymerase chain reaction (RT-qPCR) methods were used to detecte the relative level of leptin mRNA in the adipose tissue of intervenient and control groups. Their variances of fat and glucose in serum were detected with the biochemical methods. The results showed that the level of leptin mRNA of intervenient group was significantly increased (P <0.05) and the triglycered and total cholesterol were significantly decreased (P <0.05). The extracts of Polygonum cuspidatum compound could increase leptin mRNA level in adipose tissue and improve the fat metablolism in serum. However, the serum glucose of the intervenient group was a little raised (P>0.05).

[Serum sCD40L detection for risk evaluation of acute coronary syndromes].

OBJECTIVE: To investigate the value of serum soluble CD40 ligand (sCD40L) detection in risk evaluation of acute coronary syndromes (ACS). METHODS: This study involved 200 patients with established diagnosis of ACS, with death or nonfatal myocardial infarction as the end point of observation during the 6-month-long follow-up. Blood samples were obtained from the patients within the initial 72 h of ACS onset, and the levels of sCD40L and C-reactive protein (CRP) were determined with enzyme-linked immunosorbent assay (ELISA). Cardiac troponin I (cTnI) measurement was performed using chemiluminescent immunoassay. RESULTS: Of the 200 patients, 108 had serum sCD40L levels higher than 5.0 microg/L, and the levels of sCD40L, CRP and cTnI were found to significantly correlate with ACS. CONCLUSION: Independent detection of serum sCD40L, CRP and cTnI can help predict the risks of ACS, and their combined measurement may increase the sensitivity of the risk prediction and provide new cardiac makers to replace the cardiac enzymes for laboratory diagnosis and risk evaluation of cardiovascular events.

Calcium signaling events in Streptococcus pneumoniae invasion of human type II pneumocytes.

OBJECTIVE: To study whether Streptococcus pneumoniae (S.pn) can provoke filamentous actin (F-actin) rearrangements in vitro through calcium signaling pathways in type II pneumocytes(A549 cells), resulting in S.pn invasion of the cells. METHOD: After FITC-phalloidin labeling of F-actin, F-actin rearrangements were observed by S.pn adhesion to type pneumocyte A549 cells. S.pn invasion of A549 cells was determined by pretreating A549 cells with cytochalasin D. To investigate whether F-actin rearrangements could be blocked by Ca2+ inhibitors, A549 cells were pretreated with Ca2+ inhibitors dantrolene, and loaded in Fura-2/AM probe to determine the concentration of cytosolic free calcium by S.pn adhesion to A549 cells after 30, 60, and 90 min respectively. RESULTS: Intact S.pn can promote F-actin rearrangements. Cytochalasin D was able to prevent S.pn invasion of A549 cells. No invasion of A549 cell can be determined at 0.25 microg/ml of cytochalasin D. One subset of the inhibitors of Ca2+ signal transduction molecules blocked F-actin rearrangements dose-dependently, and S.pn adhesion of A549 cells for 30, 60, and 90 min increased cytosolic free calcium, reaching 487.5+/-38.1 , 548.2+/-35.6 and 557.2+/-47.5 nmol/L, respectively. They were higher than of the control group. CONCLUSION: S.pn can provoke F-actin rearrangements through Ca2+ signaling pathways, which further leads to S.pn invasion of A549 cells.