Pituitary tumor transforming gene binding factor (PBF) is required for androgen-induced prostate cancer proliferation and invasion.

Pituitary tumor transforming gene binding factor (PBF) is a proto-oncogene that plays a role in many cancers; however, its involvement in prostate cancer (PCa) remains unclear. Here, we examined PBF expression in clinical specimens and investigated its regulation and function in human PCa cell lines. Immunohistochemical staining of patient tissues revealed higher PBF expression in PCa than in benign prostatic hyperplasia or adjacent normal prostate specimens. In LNCaP and 22Rv1 cells, PBF expression was upregulated by androgen treatment in a manner partially blocked by the androgen receptor (AR) antagonist bicalutamide. We identified a novel androgen response element in the PBF gene promoter and demonstrated its functional relevance using luciferase reporter assays. Androgen treatment of LNCaP cells induced binding between the endogenous AR and the androgen response element in PBF, as measured by chromatin immunoprecipitation assays. Finally, RNA interference of PBF expression significantly reduced androgen-induced LNCaP cell growth and invasion. Thus, PBF is a novel AR target gene and plays a role in androgen-induced proliferation and metastatic functions in PCa cells.

Downregulation of microRNA-126 in endothelial progenitor cells from diabetes patients, impairs their functional properties, via target gene Spred-1.

Diabetes mellitus (DM) adversely affects the number and function of circulating endothelial progenitor cells (EPCs). Consequently, there is also a reduction in the repair mechanism of these cells, which is a critical and initiating factor in the development of diabetic vascular disease. The aim of the present study was to analyze miR expression profiles in EPCs from patients with DM and choose the most significantly regulated miR to study its possible role on EPC dysfunction and elucidate its mechanism of action. EPCs were collected from subjects with Type II DM and non-diabetic control subjects. Total RNA was harvested from EPCs, and a total of 5 candidate miRNAs were identified by microarray screening and were quantified by TaqMan real-time PCR. Lentiviral vectors expressing miR-126 and miR-126 inhibitor (anti-miR-126) were transfected into EPCs, and the EPC colony-forming capacity, proliferation activity, migratory activity, differentiation capacity, and apoptotic susceptibility were determined and Western Blotting and mRNA real-time PCR analyses were performed. To study the mechanisms, lentiviral vectors expressing Spred-1 and a short interfering RNA (siRNA) targeting Spred-1 were prepared. Five miRs were aberrantly downregulated in EPCs from DM patients. These miRs included miR-126, miR-21, miR-27a, miR-27b and miR-130a. Anti-miR-126 inhibited EPC proliferation, migration, and enhanced apoptosis. Restored miR-126 expression in EPCs from DM promoted EPC proliferation, migration, and inhibited EPC apoptosis ability. Despite this, miR-126 had no effect on EPC differentiation. miR-126 overexpression significantly downregulated Spred-1 in EPCs. The knockdown of Spred-1 expression in EPCs from DM promoted proliferation, migration, and inhibited apoptosis of the cells. The signal pathway of miR-126 effecting on EPCs is partially mediated through Ras/ERK/VEGF and PI3K/Akt/eNOS regulation. This study provides the first evidence that miR-126 is downregulated in EPCs from diabetic patients, and impairs EPCs-mediated function via its target, Spred-1, and through Ras/ERK/VEGF and PI3K/Akt/eNOS signal pathway.

[Characteristics of cervical lymph node metastasis of cN0 laryngeal carcinoma].

Objective: To investigate the incidence of occult cervical lymph node metastasis and the common neck level of metastases in cN0 laryngocarcinoma, and the relationship between the clinicopathologic features of laryngocarcinoma and cervical lymph node metastasis. Methods: A total of 506 cases with cN0 laryngocarcinoma treated at the First Affiliated Hospital of Chongqing Medical University between March 2011 and March 2018 were enrolled, and their medical records and follow-up data were retrospectively analyzed. Of them, 211 cases of were glottic carcinoma in stage T1 without neck dissection and they were observed by clinical follow-up; other 295 cases, including glottic carcinoma, supraglottic carcinoma and hypopharyngeal carcinoma in stage T2-T4 were treated with surgical resection of the primary lesions and selective neck dissection. SPSS 22.0 software was used to analyze the data. Results: The total incidence of cervical lymph node metastasis was 10.87%(55/506), with a lower incidence in T1 stage glottic carcinoma(6/211,2.84%) than that in other cases(49/295,16.61%). The incidence of cervical lymph node metastasis in glottic carcinoma (29/426, 6.81%) was lower than those in supraglottic carcinoma (22/71,30.99%) and subglottic carcinoma (4/9) (χ(2)=35.810,P<0.01).The pN+ rates of glottic carcinoma at T1, T2, T3 were 2.84%(6/211), 5.31%(6/113), 16.05%(13/81), and 19.05%(4/21), respectively (χ(2)=18.572, P<0.01). The pN+ rates of supraglottic carcinoma at T2, T3 and T4 were 3/13, 32.50%(13/40) and 6/13, respectively (χ(2)=3.649,P>0.05). The incidence of cervical lymph node metastasis in poorly differentiated carcinoma (17/42, 40.48%) was higher than those in moderately differentiated carcinoma (26/205, 12.68%) and high differentiated carcinoma(12/246, 4.88%)(χ(2)=36.356, P<0.01). Moreover, 85 pN+ lymph nodes were obtained by selective neck dissection, respectively 43(50.59%) in level Ⅱa, 30(35.29%) in level Ⅲ, 1(1.18%) in level Ⅳ and 11(12.94%) in level Ⅵ. Conclusions: The occult cervical lymph node metastasis was frequently found in cN0 laryngocarcinoma. Selective neck dissection should be performed with surgery for the primary lesions in T3-T4 glottic laryngeal cancer, T2-T4 supraglottic laryngeal cancer and subglottic carcinoma, and the neck dissection for level Ⅱa and Ⅲ is appropriate. It is required to detect pre-laryngeal and pre-tracheal lymph nodes in patients with subglottic laryngeal carcinoma.

[Extranodal NK/T-cell lymphoma, nasal type: a clinical analysis].

Objective:To explore the clinical diagnosis and treatment characteristics and clinical factors of extranodal NK/T-cell lymphoma, nasal type and provide the basis for clinical individual therapy and experience.Method:The 25 cases personal data of ENKTL received from December in 2009 to July in 2016 by our department including clinical manifestation, the serum EBV-DNA detection, imaging examination, Ann-Arbor staging, histological grade, treatment, and prognosis, etc. were retrospectively analyzed. All of the patients were pathological diagnosis and received standard, specification and system treatment. Single factor survival analysis was performed by Kaplan-Meier method and Log-rank test, and multivariate analysis was carried out using Coxproportional hazard model in the risk assessment about the factors affecting the prognosis of clinical.Result:Of the 25 patients, 15 cases(60%) were in stage ⅠE-ⅡE, which 1 year and 3 years (overall survival) OS were 100%, 100% respectively and 10 cases(40%) were in stage ⅢE-ⅣE, which 1 year and 3 years OS were respectively 40.0%, 26.7%. It had significant statistical difference (P= 0.000). Radiotherapy alone in 3 cases which 1 year and 3 years OS were respectively 100%, 100%;Chemotherapy alone in 6 cases, which 1 year and 3 years OS were 53.6% and 53.6%, respectively; 16 cases of comprehensive treatment combined radiation and chemotherapy which 1 year and 3 years OS are 84.6% and 84.6% respectively. There were significant difference between three kinds of treatment model (P= 0.027), and chemotherapy alone had the worst prognosis. Further multivariate analysis using Coxproportional hazard model showed that the course of the disease, B symptoms, EBV-DNA copy number positive, treatment mode closely associated with the prognosis (P were 0.006, 0.003, 0.010, 0.040 respectively).Conclusion:Extranodal NK/T-cell lymphoma, nasal type invasive is strong, the overall prognosis is poor. For early Ann Arbor staging, low risk and limited to the nasal cavity cases, radiotherapy alone curative effect is better. While for strong attack range or terminal patients, chemotherapy combined with radiotherapy is the first selection. In addition, this result shows that Ann Arbor staging, treatment pattern, the course of the disease, B symptoms, EBV-DNA copy number positive are independent prognostic factors.

[The relationship between the key nasal symptoms and the level of histamineand leukotriene D4 in serum and nasal secretions in allergic rhinitis].

Objective:To investigate the relationship between main nasal symptoms and the levels of histamine and leukotriene D4(LTD4) in serum and nasal secretions in allergic rhinitis(AR),and provide a preliminary guidance for individualized treatment in AR.Method:One hundred and eight cases of AR were divided into two groups,i.e.sneezing group and nasal congestion group,according to 2014 clinical guidelines for allergic rhinitis from January 2014 to June 2015.The levels of histamine and LTD4 in serum and nasal secretions were determined by enzyme-linked immunosorbent assay(ELISA) and the relationship was explored between the clinical main nasal symptoms score and the levels of histamine and LTD4.Result:The scores of sneezing(5.58±2.59)for AR were obviously related to the levels of histamine in serum(8.39±4.07)ng/ml and nasal secretion(5.06±2.47)ng/ml,(r=0.79,0.78,all P<0.05).The scores of nasal congestion(5.34±2.36) for AR were also related to the levels of LTD4 in serum(0.356±0.155 ng/ml) and nasal secretion(0.215±0.092)ng/ml,(r=0.74,0.72,all P<0.05).And the levels of histamine(8.39±4.07)ng/ml and LTD4(0.356±0.155) ng/ml in serum for AR patients were positively correlated with the levels in nasal secretions(r=0.99,P<0.01;r=0.98,P<0.01).Conclusion:In AR patients,the high levels of histamine and LTD4 in serum and nasal secretions are closely related to the sneezing symptoms and nasal obstruction symptoms,respectively.

Two-step high resolution sequence-based HLA-DRB typing of exon 2 DNA with taxonomy-based sequence analysis allele assignment.

A two-step high resolution sequence-based DRB typing method was developed. The system needs only one polymerase chain reaction (PCR) to type all functional DRB alleles of a given individual. It uses a pair of generic PCR primers to amplify exon 2 DNA of all functional DRB genes and a first-step taxonomy-based sequence analysis (FSTBSA) method to assign allele groups after sequencing the PCR products with a generic primer. In the second step, group-specific primers are used to sequence the same PCR products and a taxonomy-based sequence analysis (TBSA) is used to assign alleles. Thus, both low and high resolution DRB typing can be done with PCR amplified exon 2 DNA from a single PCR reaction. Correct allele group assignment by FSTBSA was confirmed by sequencing the PCR products with group-specific primers and correctly assigned all 158 DNA samples including 34 samples pre-typed by PCR-sequence-specific primer or PCR-sequence-specific oligonucleotide probe. FSTBSA correctly assigned 116 heterozygous combinations of 81 DRB1-DRB3/4/5 haplotypes. Sixty-seven DRB1, 6 DRB3, 1 DRB4, and 3 DRB5 alleles were identified in this study. TBSA successfully resolved all heterozygous allele combinations including 31 heterozygous combinations of 33 alleles of DRB1*03, 08, 11, 12, 13, and 14 allele groups, and six heterozygous combinations of six DRB3 alleles.

Antibodies to different members of the beta 1 (CD29) integrins induce homotypic and heterotypic cellular aggregation.

Members of the beta 1 (CD29) integrin family are involved in cellular adhesion to extracellular matrix. However, there have been several reports of CD29 integrin participation in intercellular adhesion. For example, the treatment of the human T cell line Jurkat with antibodies to alpha 4 (CD49d) causes homotypic aggregation. The present report describes the induction of aggregation of Jurkat cells by antibodies to alpha 5 (CD49e) and to a lesser extent by antibodies to the common beta 1/CD29 chain of these integrins. The metabolic requirements for these aggregations are compared with that of the CD49d-induced process. The possible involvement of fibronectin in the cytoadhesion appears to be unlikely as (1) the aggregates form in the absence of plasma fibronectin; (2) antibodies to fibronectin do not inhibit the cell adhesion; and (3) exogenous fibronectin does not influence the process. One of the cognate partners involved in the CD49e-induced aggregation appears to be present on non-antibody-treated Jurkat cells as the cells are coincorporated into aggregates of anti-CD49e-stimulated cells. The adhesion does not appear to involve members of the CD2, CD3, CD4, or CD18 receptor groups. These results indicate that interaction with alpha 4, alpha 5 chain or the beta 1 chain of the CD29 integrins leads to the induction of intercellular adhesion. The possible biological significance of this process is discussed.