OBJECTIVESTo explore whether PreS2 can change the percentage of T lymphocyte subgroups and the ration of CD4+/CD8+ in hepatocellular carcinoma (HCC) caused by HBV.

METHODSThe P120-146 region composed by the way of Merrifield, which was the most intensive antigen in PreS2 peptides, served as the antigen after dissolved in 0.01 mol/L PBS. 12 patients were chosed as the subjects, who were pathologically diagnosed as HCC after operation, were HBsAg-, HBeAg-, anti-HBc, and HBV DNA positive in serum, and expressed HBsAg in HCC tissue. The monocytes were isolated and cultured in 96 microplate with 1x 10(6) cells in every well, then the PreS2 synthetic peptides was added in at the doses of 1microg, 5microg, and 10microg, also IL-2 with 500 U was added in. Seven days later, the percentage of CD3+, CD4+, CD8+, and the ratio of CD4+/CD8+ were detected.

RESULTSIt was found that the percentage of CD4+ increased significantly (t = 3.508, P < 0.01), and the ratio of CD4+/CD8+ decreasedly obviously (t = 2.235, P < 0.05) in the 5microg PreS2 synthetic peptides group, compared with those in the control group. The percentage of CD3+ rised markedly in the IL-2 group, compared with that in the control group.

CONCLUSIONWith proper doses, PreS2 is capable of changing the expression of T lymphocyte subgroups in HCC tissue, increasing the percentage of CD4+ obviously and changing the motionless state of CD8+, to make the carcinoma cell killed through the action of CD4+ and CD8+.

Amino Acid Sequence ; CD4-CD8 Ratio ; Carcinoma, Hepatocellular ; drug therapy ; immunology ; Dose-Response Relationship, Drug ; Hepatitis B Surface Antigens ; pharmacology ; therapeutic use ; Humans ; Liver Neoplasms ; drug therapy ; immunology ; Lymphocytes, Tumor-Infiltrating ; drug effects ; immunology ; Molecular Sequence Data ; Peptide Fragments ; pharmacology ; Protein Precursors ; pharmacology ; therapeutic use

MicroRNA-21 (miR-21) is considered to play a key role in many cellular processes, affecting tumorigenesis by inhibiting target gene expression. However, its role in diffuse large B-cell lymphoma (DLBCL) is still unclear, and there are no in depth studies on relationship between miR-21 and cellular phenotype. This study was aimed to investigated the expression and role of miR-21 in the regulation of cell biological behavior in DLBCL. The expressions of miR-21 in three DLBCL cell lines were detected by real-time quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The possible roles of miR-21 in the biological and behavioral properties of DLBCL were explored by transfection of anti-miR-21 for miR-21 knockdown. In addition, PDCD4 and PTEN were assessed by luciferase reporter assay, qRT-PCR and Western blot. The results revealed that miR-21 expression was significantly upregulated in activated B-cell-like DLBCL cells as compared to germinal centre-like DLBCL cells. The inhibition of miR-21 could induce suppression of proliferation and invasion, as well as increase apoptosis in DLBCL. Moreover, knockdown of miR-21 increased the expressions of PDCD4 and PTEN at the protein level but not at the mRNA level. It is concluded the miR-21 can regulate proliferation, invasion, and apoptosis, so it has a potential therapeutic application in DLBCL.
Apoptosis ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; pathology ; MicroRNAs ; genetics ; PTEN Phosphohydrolase ; Real-Time Polymerase Chain Reaction ; Transfection ; Up-Regulation

OBJECTIVE: To explore the effect of rare synonymous variants of the ATP7B gene on the splicing of its precursor mRNA. METHODS: A total of 248 rare synonymous variants with allelic frequency of <0.005 were retrieved from the ExAc database. Human Splicing Finder (HSF) was used to predict their effect on the splicing of precursor mRNA. And ESE Finder 3.0 was used to predict the effect of such variants on the binding ability of SR protein family. Rare synonymous variants affecting the binding of two or more SR proteins were selected and verified with an in vitro mini gene splicing report system. RESULTS: HSF analysis indicated that 136 of the 248 rare synonymous variants may destroy the exonic splicing enhancer (ESE) motif. Analysis using ESE Finder 3.0 indicated that 19 of them may affect the binding of two or more SR proteins at the same time. In vitro mini gene experiment confirmed that the c.1620C>T (p.L540L) and c.3888C>T (p.A1296A) variants could lead to abnormal splicing of the corresponding exons, resulting in complete skipping of exon 4 and 25% increase in the skipping of exon 18, respectively. CONCLUSION Synonymous variants may affect the splicing of precursor mRNA in various ways, particularly the destruction of ESE motif. This study confirmed that the c.1620C>T (p.L540L) and c.3888C>T (p.A1296A) variants can affect the mRNA splicing of the ATP7B gene, resulting in skipping of corresponding exons, which may provide a basis for genetic diagnosis and consultation of carriers.
Alternative Splicing ; Copper-Transporting ATPases/genetics* ; Enhancer Elements, Genetic ; Exons ; Gene Frequency ; Humans ; RNA, Messenger/genetics*

Aromatic inhalation therapy is a key traditional Chinese medicine （TCM） approach for preventing respiratory infectious diseases. Its foundational theory， "aromatics acting on the spleen"， is deeply rooted in TCM principles and supported by modern medical research. The theory posits that the aromatic properties of medicinals primarily act on the spleen， and the aromatic inhalation therapy achieved its protective effects by modulation of the spleen and spleen channel to enhance the regulation of wei qi， striae and interstices. In TCM， the spleen is considered the mother of the lungs， with the function of nurturing lung； it is also seen as the source of wei qi， responsible for external defense； and the root of healthy qi， forming the foundation of acquired （postnatal） constitution. Thus， preventive strategies for respiratory infectious diseases focus on strengthening the spleen. From a modern medical perspective， the spleen's role in regulating lung immune responses， the shared immune functions of the respiratory and gastrointestinal mucosa， and the spleen's overall immune modulation provide scientific evidence for using aromatic inhalation therapy to prevent respiratory infections. Additionally， aromatic inhalation therapy offers several advantages， including direct action， rapid onset， minimal side effects， controllable risks， convenience， and ease of dissemination， making it a practical and effective preventive measure for respiratory infectious diseases.

Macrophages play an important role in material-related immune responses and bone formation, but the functionality of macrophage-derived extracellular vesicles (EVs) in material-mediated bone regeneration is still unclear. Here, we evaluated intracellular communication through small extracellular vesicles (sEVs) and its effects on endogenous bone regeneration mediated by biomimetic intrafibrillarly mineralized collagen (IMC). After implantation in the bone defect area, IMC generated more neobone and recruited more mesenchymal stem cells (MSCs) than did extrafibrillarly mineralized collagen (EMC). More CD63
Biomimetics ; Bone Regeneration ; Cell Differentiation ; Collagen ; Extracellular Vesicles ; Macrophages ; Osteogenesis