Abstract. Antigen targeted immunotherapies might represent a novel treatment for B-cell chronic lymphocytic leukemia (B-CLL). We screened the mRNA expression of tumorassociated antigens (TAAs) from the literature (fibromodulin, survivin, OFA-iLRP, BAGE, G250, MAGE1, PRAME, proteinase, syntaxin, hTERT, WT-1) and TAAs defined previously by serological analysis of cDNA expression libraries from leukemic cells (PINCH, HSJ2, MAZ, MPP11, RHAMM/CD168, NY-Ren60). Peripheral blood mononuclear cells from 43 B-CLL patients and 20 healthy volunteers (HVs) were examined by conventional and quantitative RT-PCR. mRNA of RHAMM/CD168, fibromodulin, syntaxin and NYRen60 was expressed in 55-90%, and mRNA of HSJ2, MAZ and OFAiLRP was expressed in 90-100% of the patients. No expression of WT-1, hTERT, BAGE, G250, MAGE1 or survivin was observed. Low (2-20%) expression frequencies of MPP11, PINCH, PRAME and proteinase were detected. RHAMM/CD168, fibromodulin, PRAME and MPP11 showed expression in B-CLL patients, but not in HVs. Because of the exquisite tissue expression of RHAMM/CD168 and its high expression frequency in CLL patients, mixed lymphocyte peptide culture (MLPC), enzyme-linked immunosorbent spot (ELISPOT) and flow cytometry were performed for antigen specific T-cells. In MLPC, RHAMM specific responses by CD8 + HLA-A2/R3tetramer + CCR7-CD45RA high effector T-cells were detected. RHAMM/CD168 might be a possible target for future immunotherapies in both ZAP-70(+) and ZAP-70(-) B-CLL patients.

Aging salivary glands are characterized by a reduced volume of acini, increased duct volume, decline in the rate of synthesis of proteins and their mRNA, and decreased saliva flow. Sialin is a versatile anion transporter that is highly expressed in salivary glands and may also participate in maintenance of salivary gland function. We investigated age-associated sialin expression in salivary glands. Submandibular glands of mice and human parotid gland were collected at different ages. Western blot and real-time reverse transcription-polymerase chain reaction were used to analyze the protein and mRNA expression levels of sialin, respectively. Immunohistochemical and immunofluorescence staining were used to evaluate the histological pattern of sialin expression. We found that the protein and mRNA expression levels of sialin decreased with aging. The expression of sialin in the striated and excretory ducts of both mouse and human glands were reduced with age. In the mouse glands, the basal 2/3 of the cytoplasm was stained, while in the human glands the stain extended to the luminal surface. Sialin was expressed in the basal cytoplasmic membrane of acini and in some myoepithelial-like cells in young salivary glands, but had nearly disappeared in the aged salivary glands. Changes in sialin expression may be associated with changes in physiological function of salivary glands with aging.

The purpose of this study was to identify potential miRNAs and mRNAs involved in chemotherapy insensitivity in hypopharyngeal squamous cell carcinoma (HSCC) and to explore the underlying mechanisms involved to provide diagnostic markers and therapeutic targets for HSCC. We used microarrays to identify differences in both the mRNA and miRNA expression profiles between a group (twelve patients) sensitive to chemotherapy and a resistant group (nine patients). We then employed bioinformatics tools to examine the functions and pathways involved. The genes and miRNAs most related to chemotherapy sensitivity in HSCC were screened. Finally, a miRNA-mRNA-phenotype network was constructed with an integrated analysis based on the identified miRNAs and mRNAs. Nine differentially expressed miRNAs and one hundred differentially expressed mRNAs were identified, and the functions of these genes and miRNAs were predicted. Bioinformatics analysis revealed a regulatory network consisting of eight genes and two miRNAs that influenced HSCC chemosensitivity. According to our analysis, CCL4L1 may be a potential molecular marker for HSCC chemotherapy, and excess CCL4L1 leads to the upregulation of PRAME and the downregulation of miR-375, thus decreasing HSPB8 expression and promoting chemotherapy sensitivity. Our work provides reliable data for further studies investigating the mechanism of HSCC chemotherapy sensitivity.

Although endoplasmic reticulum (ER) stress is critical in various liver diseases, its role in acute-on-chronic liver failure (AoCLF) caused by acute exacerbation of chronic hepatitis B (CHB) is still elusive. This study aimed to analyse ER stress responses in the progression of HBV-related AoCLF. Normal liver tissues (n = 10), liver tissues of CHB (n = 12) and HBV-related patients with AoCLF (n = 19) were used. Electron microscopy of the ultrastructure of the ER was carried out on liver specimens. The gene and protein expression levels of ER stress-related genes were measured. We further analysed the correlation between the expression levels of ER stress-related molecules and liver injury. Electron microscopy identified typical features of the ER microstructure in AoCLF subjects. Among the three pathways of unfolded protein responses, the PKR-like ER kinase and inositol-requiring enzyme 1 signalling pathway were activated in CHB subjects and inactivated in AoCLF subjects, while the activating transcription factor 6 signalling pathway was sustained in the activated form during the progression of AoCLF; the expression of glucose-regulated protein (Grp)78 and Grp94 was gradually decreased in AoCLF subjects compared to healthy individuals and CHB subjects, showing a negative correlation with serum ALT, AST and TBIL; moreover, the ER stress-related apoptosis molecules were activated in the progression of acute exacerbation of CHB. The dysregulated ER stress response may play a complicated role in the pathogenesis of AoCLF, and a severe ER stress response may predict the occurrence of AoCLF caused by acute exacerbation of CHB.

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