LncRNA THAP9-AS1 accelerates cell growth of esophageal squamous cell carcinoma through sponging miR-335-5p to regulate SGMS2.

Esophageal squamous cell carcinoma (ESCC) is kind of common and aggressive malignant tumors with high incidence and mortality all over the world. Accumulating studies have reported that long non-coding RNAs (lncRNAs) can play a vital regulatory role in human cancers. THAP9 antisense RNA 1 (THAP9-AS1) has been identified as an oncogene in several cancers. But its role in ESCC remains to be studied. In our research, THAP9-AS1 expression in ESCC cell lines was analyzed by real-time quantitative polymerase chain reaction (RT-qPCR). Cell proliferation, migration, invasion and apoptosis as well as EMT process were analyzed by 5-Ethynyl-2'-deoxyuridine ( EdU), Transwell, Terminal deoxynucleotidyl transferase dUTP Nick-End Labeling (TUNEL) and western blot experiments. The interplay of THAP9-AS1, miR-335-5p and sphingomyelin synthase 2 (SGMS2) was analyzed by luciferase reporter assay and RNA immunoprecipitation (RIP) assay. We discovered that THAP9-AS1 was highly expressed in ESCC cell lines and that the knockdown of THAP9-AS1 inhibited proliferation, migration, and invasion as well as EMT of ECSS cells but enhanced cell apoptosis. Furthermore, miR-335-5p was proved to be sponged by THAP9-AS1 and its up-regulation could repress ESCC progression. Additionally, SGMS2 was verified to be the target gene of miR-335-5p. In rescue assay, SGMS2 overexpression could offset the suppressive role of THAP9-AS1 depletion on ESCC progression. In short, THAP9-AS1 accelerated cell growth of ESCC through sponging miR-335-5p to regulate SGMS2.

Efficacy of Sublingual Administration of Dermatophagoides Farinae Drops for Treatment of Pediatric Allergic Rhinitis Accompanied by Adenoid Hypertrophy and Improvement of Immune Function.

BACKGROUND The aim of this study was to determine the efficacy of sublingual administration of Dermatophagoides farinae drops for the treatment of allergic rhinitis (AR) accompanied by adenoid hypertrophy and the effect on immune function in children. MATERIAL AND METHODS Eosinophil counts in peripheral blood before and after treatment were determined; serum levels of immunoglobulin E (IgE), total IgE (T-IgE), immunoglobulin G4 (IgG4), interleukin-2 (IL-2), and interleukin-6 (IL-6) before and after treatment were detected by enzyme-linked immunosorbent assay. RESULTS The total effective rate in the study group was significantly higher than that in the control group (P<0.05). In both the study and control groups, symptom scores, medication scores, eosinophil counts in the peripheral blood, and serum levels of IgE, T-IgE, and IL-6 were significantly lower than those before treatment (P<0.05), while the serum levels of IgG4 and IL-2 were significantly higher than those before treatment (P<0.05). After treatment, symptom scores, medication scores, eosinophil counts in the peripheral blood, and serum levels of IgE, T-IgE, and IL-6 in the study group were significantly lower than those in the control group (P<0.05), while the serum levels of IgG4 and IL-2 were significantly higher in the study group than those in the control group (P<0.05). CONCLUSIONS Sublingual administration of D. farinae drops improved the clinical symptoms of pediatric AR caused by Dermatophagoides mites and improved the immune functions in children.

Diverse Changes of Circulating Fibroblast Growth Factor 21 Levels in Hepatitis B Virus-Related Diseases.

Fibroblast growth factor 21 (FGF21), a stress-induced hormone in the liver, has been shown the protective functions in pathological conditions. The study investigated the association of circulating FGF21 with hepatitis B virus (HBV) infection and its related diseases. Serum FGF21 levels were measured in 33 acute hepatitis B (AHB), 75 chronic hepatitis B (CHB) and 66 CHB patients with advanced liver diseases including liver cirrhosis, acute-on-chronic liver failure (ALCF) and hepatocellular carcinoma (HCC) together with 200 age- and BMI-matched healthy controls. FGF21 levels were significantly increased in AHB patients and rapidly returned to normal levels after treatment. FGF21 levels reflected the degree of liver injury caused by AHB. However, serum FGF21 levels were decreased in CHB patients especially in those who developed cirrhosis and were associated with hepatic protein synthesis capacity. Serum FGF21 in CHB patients were increased with the occurrence of ACLF. Notably, in CHB patients who developed HCC, serum FGF21 exhibited a dramatic increase, which may provide important information on monitoring tumorigenesis in CHB patients. In conclusion, we revealed the diverse changes of circulating FGF21 in HBV-related diseases. FGF21 may be a useful biomarker in monitoring the tumorigenesis in patients with CHB.

[One case of laryngopharyngeal recess fibroma].

To report a case of laryngopharyngeal recess fibroma with the clinical and pathological characteristics. The laryngopharyngeal recess neoplasm was expect with pedestal laryngoscope. The postoperative pathologic diagnosis was laryngopharyngeal recess fibroma. The tumor did not recurred after one year following-up. Surgery is the first selection for treatment of patient with laryngopharyngeal recess neoplasm. A closed follow-up is needed.

Lentivirus-mediated silencing of rhomboid domain containing 1 suppresses tumor growth and induces apoptosis in hepatoma HepG2 cells.

Rhomboids were identified as the first intramembrane serine proteases about 10 years ago. Since then, the study of the rhomboid protease family has blossomed. Rhomboid domain containing 1 (RHBDD1), highly- expressed in human testis, contains a rhomboid domain with unknown function. In the present study, we tested the hypothesis that RHBDD1 was associated with proliferation and apoptosis in hepatocellular carcinoma using recombinant lentivirus-mediated silencing of RHBDD1 in HepG2 cells. Our results showed that down-regulation of RHBDD1 mRNA levels markedly suppressed proliferation and colony formation capacity of HepG2 human hepatoma cancer cells in vitro, and induced cell cycle arrest. We also found that RHBDD1 silencing could obviously trigger HepG2 cell apoptosis. In summary, it was demonstrated that RHBDD1 might be a positive regulator for proliferative and apoptotic characteristics of hepatocellular carcinoma.

Intracellular-delivery of a single-chain antibody against hepatitis B core protein via cell-penetrating peptide inhibits hepatitis B virus replication in vitro.

Assembly of nucleocapsids is an attractive target for novel anti-hepatitis B virus (HBV) agents, and intracellular single-chain variable fragment (scFv) antibodies against HBV core (HBc) protein are a class of potential alternatives for this purpose; however, their application is limited by the lack of a suitable means of delivery. Owing to the favorable performance of cytoplasmic transduction peptide (CTP) in cargo delivery in hepatocytes, we purified an anti-HBc scFv fused to CTP using a previous screened sequence by a prokaryotic expression system and evaluated its efficacy in the inhibition of HBV in vitro. Our results showed that cytoplasmic translocation of the previous anti-HBc scFv was achieved by CTP in HepG2.2.15 cells. Immunoprecipitation analysis indicated the fusion protein anti-HBc scFv-CTP interacted with its target antigen HBc, and negligible cytotoxicity was observed. Moreover, the anti-HBc scFv-CTP interfered with nucleocapsid assembly and markedly reduced both the supernatant HBV DNA level and the intracellular DNA replication intermediates, with a 5.1 µM of half maximal effect concentration and a dose-dependent effect. In conclusion, this novel anti-HBc scFv fused to CTP demonstrated inhibitory activity of HBV replication in vitro and warrants further in vivo study.

[Clinical analysis of primary laryngeal amyloidosis].

OBJECTIVE: To analyze the clinical features and treatment protocols of primary laryngeal amyloidosis. METHOD: Retrospective study of 5 patient hospitalized from 1996 - 2011. All of the patients by resection lesions, including four routine throat tumor resection, and burst throat by supporting laryngoscope in 1 case, all did not give lesions resection radiation and hormone therapy. RESULT: All the 5 patients recovered clinically. There were 3 patients followed up for 0.3-7.5 years with a mean time of 3.3 years without recurrence, 2 patients lost follow-up. CONCLUSION: Middle ages seemed to be more vulnerable. The most common disease region is true vocal cord, followed by false vocal cord, epiglottis former clearance etc. Early surgical treatment of this disease is the most important treatment, larynx endoscopic and CT for the diagnosis of great value, and pathologic biopsy especially Congo red stain positive is the basis of the specific diagnosis of this disease.

Inhibition of Csn3 expression induces growth arrest and apoptosis of hepatocellular carcinoma cells.

PURPOSE: Csn3 (or CSN3) encodes the third subunit of an eight-subunit complex, the COP9 signalosome (CSN), which acts as a protein kinase and a deneddylase in mammalian cells. Previous studies have shown that Csn3 is essential for maintenance of cell proliferation in the mouse embryonic epiblast and associated with the tumorigenesis process in osteosarcoma. However, its correlation with hepatocellular carcinoma (HCC) has not been explored yet. METHODS: The expression of Csn3 in HCC (n = 30), cirrhosis (n = 30), and normal tissues (n = 30) was detected using immunohistochemical analysis. The impacts of lentivirus-mediated inhibition of Csn3 on HCC cells were detected using MTT, BrdU incorporation assay, and flow cytometric analysis. In addition, the colony formation and tumor growth ability in nude mice were detected to define the role of Csn3 in tumorigenesis. RESULTS: Knockdown of Csn3 expression in HCC cell lines (SMMC-7721 and Hep3B) significantly inhibits the tumor growth both in vitro and in vivo. Further investigation indicates that this growth inhibition effect may be mediated through cell cycle arrest in G0/G1 phase and inductions of pro-apoptotic proteins BIK and Caspase-8. In addition, knockdown of Csn3 expression evidently suppresses tumor growth in a xenograft nude mice model. CONCLUSION: Collectively, this study demonstrates Csn3 as an oncogene that regulates the tumorigenesis process in HCC cells.

[HBcAg-specific CTL response induced by PTD-HBcAg fusion protein to inhibit HBV replication in vitro].

AIM: To investigate the effects of specific cytotoxic T lymphocyte induced by sensitized dendritic cells(DCs)with PTD-HBcAg fusion protein on inhibiting HBV in vitro. METHODS: DCs were cultured and induced maturation by different fusion proteins, and cocultured with allogeneic T cells to detect the secretion level of IL-2, IL-4, IL-10 and INF-gamma in the supernatants of T cells by ELISA. Intracellular cytokine of proliferative T cells was analyzed by flow cytometry and the specific CTL activity was measured by a lactate dehydrogenase (LDH) release assay. The levels of HBsAg and HBV DNA in the supernatant of HepG2.2.15 cells were detected. RESULTS: Sensitized dendritic cells by different fusion proteins could promote cytokine secretion effectively, the levels of IL-2(552.7+/-117.5 ng/L) and INF-gamma(150.6+/-7.945 ng/L)induced by M-PTD-HBcAg were higher than that induced by M-HBcAg (420+/-12.47 ng/L and 107.5+/-12.19 ng/L, respectively).The amount of CTLs induced by PTD-HBcAg fusion protein was more than others by the analysis of intracellular cytokine of proliferative T cells. Specific CTL killing activity could be induced by PTD-HBcAg and HBcAg(P<0.05) and PTD-HBcAg significantly decreased the levels of HBsAg and HBV DNA. CONCLUSION: Sensitised DCs by PTD-HBcAg fusion protein can stimulate cytokine secretion and increase cytotoxic T lymphocytes generation effectively, and also enhance the specific CTL activity to decrease the level of HBsAg and HBV DNA in supernatants of HepG2.2.15 cells.

Enhancement of cytotoxic T lymphocyte activity by dendritic cells loaded with Tat-protein transduction domain-fused hepatitis B virus core antigen.

The protein transduction domain (PTD) of human immunodeficiency virus-1-Tat protein has a unique potency to penetrate the cellular membranes. To synthesize the sequence of Tat-PTD47-57 and hepatitis B virus core antigen (HBcAg), we spliced these sequences and linked a fusion gene into the pMAL-c2x vector. The fusion proteins were purified by affinity chromatography and pulsed with bone marrow-derived dendritic cells (DCs), and the transduction of recombinant protein was detected by immunofluorescence antibody assay. Results showed that recombinant PTD-HBcAg could penetrate into DC cytoplasm while recombinant HBcAg was detected on the surface of cells. The percentage of DC surface molecules, such as CD80, CD86 and major histocompatibility complex II, and production of cytokine (IL-12p70) induced by recombinant PTD-HBcAg were significantly higher than those induced by recombinant HBcAg or tumor necrosis factor-alpha. DCs treated with PTD-HBcAg induced T cells to differentiate into specific cytotoxic T lymphocytes (CTLs) and enhanced the CTL killing response. In conclusion, the expressed and purified PTD-HBcAg fusion protein could penetrate into cells through the plasma membrane, promote DC maturation, and enhance T cells response to generate HBcAg-specific CTLs efficiently.