Mesenchymal phenotype of circulating tumor cells is associated with distant metastasis in breast cancer patients.

In this study, we investigated the relationship between the epithelial-mesenchymal transition phenotype of circulating tumor cells (CTCs) and distant metastasis in breast cancer patients. We analyzed the expression of epithelial (epithelial cell adhesion molecule, cytokeratin [CK]8, CK18 and CK19) and mesenchymal (vimentin and TWIST1) markers in CTCs from a large cohort of Chinese breast cancer patients (N=1083) using Canpatrol™ CTC assays. We identified CTCs in 84.9% (920/1083) of the breast cancer patients enrolled in this study. Among these 920 patients, 547 showed epithelial CTCs, 793 showed biphenotypic CTCs and 516 showed mesenchymal CTCs. Receiver operating characteristic (ROC) curves demonstrated circulation of both biphenotypic and mesenchymal CTCs (area under ROC curve value: 0.728; sensitivity: 68.7% and specificity: 71.6%) in patients was associated with distant metastasis. These findings demonstrate that the epithelial-mesenchymal transition phenotype of CTCs is a potential biomarker predictive of distant metastasis in breast cancer.

Preliminary screening of type IV secretion system in divergent geographic sources of Clostridium difficile.

In the present study, different geographical sources and sequence types (STs) of Clostridium difficile were preliminarily screened to investigate the distribution profiles of three core genes, VirB4, VirB6 and VirD4, of the type IV secretion system (T4SS). A total of 37 C. difficile strains from different sources were screened, inoculated and prepared for genome extraction. C. difficile toxins A and B were subjected to identification and multilocus sequence typing (MLST) analysis. The T4SS gene then underwent polymerase chain reaction amplification and sequencing analysis. Of the 37 strains, 25 were toxin A- and toxin B-positive, and 12 were toxin A-negative and toxin B-positive. MLST detected 11 strains with ST37, 10 with ST2, 6 with ST35, 7 with ST3, 1 with ST54, 1 with ST1 and 1 with ST119. The detection rates of VirB4, VirB6 and VirD4 were all 100% in colonies exhibiting T4SS. Single nucleotide polymorphisms (SNPs) were detected in a minority of strains. C. difficile strains with identical STs shared the same SNP loci for T4SS, and those with different STs had different SNP loci. The results of the present study may provide evidence for subsequent identification of T4SS distribution, epidemiological investigations, polymorphism analyses and research into the association between T4SS, cytotoxicity and enterotoxication in C. difficile.

Evolution of correlation between Helicobacter pylori infection and autoimmune liver disease.

The present study was planned to evaluate correlation between Helicobacter pylori (HP) infection and autoimmune liver disease (AILD). A total of 60 patients diagnosed with AILD in Affiliated Hospital of Binzhou Medical College were continuously enrolled in the present study. HP infection was detected by 13C-urea breath test. The levels of anti-myeloperoxidase were tested by ELISA. The positive rate of anti-nuclear antibody (ANA), anti-mitochondrial antibody (AMA), anti-smooth muscle antibody (SMA) and anti-neutrophil cytoplasm antibody (ANCA) were tested by indirect immunofluorescence. The positive rates of anti-mitochondrial antibody (AMA-M2), anti-liver-kidney microsomal antibody (LKM-1), anti-liver cytoplasm antibody I (LC-1) and anti-soluble liver antigen/liver-pancreas antigen (SLA/LP) were tested by immunoblotting. Liver function indexes including alanine transaminase, aspartate transaminase, alkaline phosphatase and glutamyltransferase, were analyzed with a fully automatic biochemical analyzer. The levels of serum cytokine IFN-γ, interleukin-6 (IL-6), IL-10 and tumor necrosis factor-α (TNF-α) were tested by ELISA. A total of 37 patients (61.67%) were observed to be HP-positive. MPO-positive rate, positive rate of ANA, AMA, SMA and ANCA and positive rate of AMA-M2, LKM-1, LC-1 and SLA/LP in patients with positive HP infection were significantly higher than those of patients with negative HP infection. On the other hand, the levels of liver function indices did not showed any significant differences among HP-positive cases or HP-negative cases. However, the levels of IFN-γ, IL-6, IL-10 and TNF-α in patients with positive HP infection were significantly higher than those of patients with negative HP infection. In conclusion, the positive infection rate of HP infection in patients with AILD is high and is closely associated with various positive immune antibodies as well as cytokine levels.

Evaluation of Glutathione Peroxidase 4 role in Preeclampsia.

Preeclampsia (PE) is a pregnancy-specific syndrome that may be lifethreatening to pregnancies and fetus. Glutathione Peroxidase 4 (GPx4) is a powerful antioxidant enzyme that can provide protection from oxidative stress damage which plays a pivotal role in the pathology of PE. Therefore, this study aims to investigate the association between Gpx4 polymorphisms and the susceptibility to PE in Chinese Han women. TaqMan allelic discrimination real-time PCR was used to perform the genotyping of rs713041 and rs4807542 in 1008 PE patients and 1386 normotensive pregnancies. Obviously statistical difference of genotypic and allelic frequencies were found of rs713041 in GPx4 between PE patients and controls and the C allele has the higher risk for pathogenesis of PE (χ(2) = 12.292, P = 0.002 by genotype; χ(2) = 11.035, P = 0.001, OR = 1.216, 95% CI 1.084-1.365 by allele). Additionally, when subdividing these samples into CC + CT and TT groups, we found a significant difference between the two groups (χ(2) = 11.241, P = 0.001, OR = 1.417, 95% CI 1.155-1.738). Furthermore, the genotype of rs713041 was found to be associated with the mild, severe and early-onset PE. Our results suggest that rs713041 in GPx4 may play a key role in the pathogenesis of PE.

Immune Reconstitution of CD4+T Cells after Allogeneic Hematopoietic Stem Cell Transplantation and its Correlation with Invasive Fungal Infection in Patients with Hematological Malignancies.

OBJECTIVE: To explore the immune reconstitution of CD4+T cells after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) and its relationship with invasive fungal infection (IFI) in patients with hematological malignancies. MATERIALS AND METHODS: Forty-seven patients with hematological malignancies undergoing Allo- HSCT in Binzhou Medical University Hospital from February, 2010 to October, 2014 were selected. At 1, 2 and 3 months after transplantation, the immune subpopulations and concentration of cytokines were assessed respectively using flow cytometry (FCM) and enzyme linked immunosorbent assay (ELISA). The incidence of IFI after transplantation and its correlation with immune reconstitution of CD4+T cells were investigated. RESULTS: The number of CD4+T cells and immune subpopulations increased progressively after transplantation as time went on, but the subpopulation cell count 3 months after transplantation was still significantly lower than in the control group (p<0.01). In comparison to the control group, the levels of interleukin-6 (IL-6) and IL-10 after transplantation rose evidently (p<0.01), while that of transforming growth factor-ß (TGF-ß) was decreased (p<0.01). There was no statistically significant difference level of interferon-γ (IFN-γ) (p>0.05). The incidence of IFI was 19.2% (9/47), and multivariate logistic regression revealed that IFI might be related to Th17 cell count (p<0.05), instead of Th1, Th2 and Treg cell counts as well as IL-6, IL-10, TGF-ß and IFN-γ levels (p>0.05). CONCLUSIONS: After Allo-HSCT, the immune reconstitution of CD4+T cells is delayed and Th17 cell count decreases obviously, which may be related to occurrence of IFI.

MicroRNAs as a novel class of diagnostic biomarkers in detection of hepatocellular carcinoma: a meta-analysis.

MicroRNAs (miRNAs) have been proposed as promising diagnostic biomarkers for many diseases, particularly in the field of cancer research. Numerous studies have explored the use of miRNAs in the detection of hepatocellular carcinoma (HCC), with some reporting inconsistent results. Thus, we conducted this meta-analysis to evaluate the potential diagnostic value of miRNAs in HCC. All relevant literature was collected from the PubMed and other databases before June 3, 2014. The summary receiver operator characteristic (SROC) curve and other parameters were used to estimate overall predictive performance. Fourteen studies involving 1,848 cases with HCC and 1,187 controls (576 healthy controls and 611 individuals with chronic liver diseases) were included in this meta-analysis. SROC analyses for the diagnostic power of miRNAs yielded an area under the curve (AUC) of 0.93 with 86 % sensitivity and 86 % specificity in discriminating patients with HCC from healthy subjects and an AUC of 0.88 with 79 % sensitivity and 83 % specificity in differentiating patients with HCC from those with chronic liver diseases (CLDs). Furthermore, subgroup analyses showed that miRNA panels yielded excellent diagnostic characteristics, with an AUC of 0.99 (96 % sensitivity and 96 % specificity) for detection of HCC from healthy controls and an AUC of 0.93 (85 % sensitivity and 88 % specificity) for HCC from those with CLDs. MiRNAs might be novel potential biomarkers for the diagnosis of HCC, and a combination of multiple miRNAs could significantly improve the diagnostic accuracy.