Objective To evaluate the anti-cervical carcinoma cells effects of oncolytic adenovirus d/1520 and Ad-Endo in vitro,and investigate the influence of d/1520 on the Endostatin expression directed by Ad-Endo in cervical carcinoma cells.Methods After cervical carcinoma cells were infected with d/1520 or/and Ad-Endo,the cytopathic effects (CPE) were observed,and the cytotoxicities were measured by MTT assay.Endestatin concentrations were measured by ELISA.Results d/1520 can replicate in cervical carcinoma cells and induce cytopathie effects; In MTT assay,Ad-Endo hardly inhibited the growth of cervical carcinoma cells,while d/1520 inhibited efficiently the proliferation of cervical cancer cells.And the inhibitory rates in Ad-Endo + d/1520 group were higher than that in d/1520 group.Ad-Endo expressed Endestatin protein in cervical cancer cells,while Endostatin concentrations in Ad-Endo+dl1520 group were much higer than those in Ad-Endo group.Conclusions dl1520 can kill cervical carcinoma cells in vitro,and this eytotoxieity can be enhanced by combining with Ad-Endo.The expression of Endostatin directed by Ad-Endo in cervical cancer cells can be dramatically increased by dl1520.

Purpose To evaluate the diagnostic values of Clusterin, CXCL13, Podoplanin (D2-40), CD21 and CD35 in follcular den-dritic cell sarcoma. Methods The expression levels of 10 cases of follcular dendritic cell sarcoma ( FDCS) and 12 types of FDCS mimics (83 cases in total) were investigated by immunohistochemical methods, the latter including solitary fibrous tumor, leiomyosar-coma, gastrointestinal stromal tumor and others. The diagnostic validities of the five biomarkers were compared. Results ( 1 ) The positive rates of Clusterin, CXCL13, D2-40, CD21 and CD35 in the FDCS group were 100%, 70%, 60%, 90% and 80%, respec-tively, the corresponding rates in the control group were 30%, 4%, 11%, 2% and 0 in turn. (2) The five biomarkers could be cate-gorized into 3 groups, according to their diagnostic values in FDCS. The first group included CD21 and CD35, which had much higher sensitivities (90%, 80%), specificities (100%, 98%) and accuracies (98%, 96%), compared with the other biomarkers. The second group included CXCL13 and D2-40, which had a relatively lower sensitivities (70%, 60%), specificities (96%, 89%) and accuracies (94%, 86%), compared with CD21 and CD35. The third group included Clusterin, which had the highest sensitivity (100%), while the specificity (70%)and accuracy (73%) were inferior to the first and second groups. (3) The diagnostic values of CD21 and CD35 combination were 100%, 98%, 98%, 83% and 100%, respectively. Conclusions (1) CD21 and CD35 are the most valuable biomarkers for FDCS. The combined diagnostic effect of the two markers is generally superior to that of single marker. (2) Clusterin has the highest sensitivity for FDCS. However, the frequent expression in FDCS mimickers restricts its diagnostic values for FDCS. (3) CXCL13 and D2-40 may be used as a marker for FDCS, but are not recommended as the first selection based on their diagnostic performance. (4) On the reasons that FDCS mimickers may not uncommonly express Clusterin and D2-40, and rarely show positivity for CD21 or CXCL13, more attention should be paid to the phenomenon to avoid misdiagnosis.

Purpose To detect high mobility group protein A2 (HMGA2) expression in breast cancer, and to analyze its relationship with clinicopathological features and the levels of HMGA2 in different molecular subtypes of breast carcinomas. Methods An immu-nohistochemical study was undertaken for measuring the levels of HMGA2 in 58 breast carcinomas. Results ( 1 ) The expression of HMGA2 was 0, 62. 5%, 60. 0%, 100. 0% and 80. 0% in Lum A, Lum B, HER-2-OE, basal-like breast carcinoma (BLBC) and un-classification phenotype respectively ( triple negative breast cancer, 92. 3%) . High expression of HMGA2 was associated with the tri-ple-negative breast cancer (TNBC) subtypes (P<0. 01). (2) An association was identified between high expression of HMGA2, and high tumor grade, lymph node metastasis (P<0. 05), positive Ki-67, CK5/6 and EGFR (P<0. 05), negative ER and PR (P<0. 01), but no association was observed for tumor size and patients’age. Conclusion An association is identified between high ex-pression, and high tumor grade, lymph node metastasis, positive Ki-67, EGFR and CK5/6, negative ER and PR, that means a high expression of HMGA2 is associated with an adverse outcome in breast cancer. High expression of HMGA2 are associated with the TNBC subtypes. Thus recognizing HMGA2 as a rational target in TNBC. The results of the study have implications for therapeutic target iden-tification and the design of future clinical trials for TNBC.

Purpose To study the levels and subcellular localization of β-catenin in 5 different molecular subtypes of breast carcino-mas. Methods An immunohistochemical study was undertaken for measuring the levels and subcellular localization ofβ-catenin in 58 breast carcinomas. Results ( 1 ) The cytoplasmic expression of β-catenin was 21. 1%, 50%, 60%, 100% and 60% ( TNBC 84. 6%) in Lumina A, Lumina B, HER-2-OE, basal-like breast carcinoma ( BLBC) and uncl phenotype respectively. High cytoplas-mic expression was associated with the BLBC and TNBC subtypes ( P<0. 05 ) . ( 2 ) An association was identified between high cyto-plasmic expression of β-catenin, and high tumor grade (P<0. 01), abnormal E-cadherin, positive Ki-67 and CK5/6 (P<0. 05), negative ER and PR (P<0. 01), but no association was observed for lymph node metastasis, tumor size and patients’age. Conclu-sion An association is identified between high cytoplasmic expression, and high tumor grade, positive Ki-67 and CK5/6, negative ER and PR, that means a high cytoplasmic expression ofβ-catenin is associated with an adverse outcome in breast cancer. High cytoplas-mic expression are associated with the BLBC and TNBC subtypes whcih recognizing Wnt signaling as a rational target in TNBC and BLBC. The results of the study have implications for therapeutic target identification and the design of future clinical trials for TNBC and BLBC.

Objective To explore the value of bone marrow biopsy ( BMB) in early diagnosis of aggressive NK cell leukaemia( ANKL). Methods The clinical data of ten cases with ANKL were retrospectively analyzed,morphology, immunophenotype and hybridization in situ of bone marrow were analyzed. Results In all cases, BMB showed hypercellular, with 4 cases markedly hypercellular. Atypical neoplastic cells demonstrated focal and fascicle growth pattern and were composed of median-sized cells with a few cytoplasm, slightly irregular nuclei, fine chromatin, indistinct nucleoli and some mitotic counting. Characteristic histocytes with haemophagocytosis were observed in the bone marrow slides. The neoplastic cells were positive for CD2、CD3e、CD7、CD56、TIA-1. EBER was found positive in all cases. Conclusion Bone marrow biopsy and immunochemistry were essential and reliable diagnostic tool in early diagnosis of ANKL.

Objective To investigate the expression of transforming growth factor beta (TGF-β) and Smad signaling in ankylosing spondylitis (AS) and to explore their roles in the pathogenesis. Methods Fiftythree patients with AS were included in the study. In these 53 cases, 30 patients were performed computed tomography-guided needle biopsy in sacroiliac joint. Serum TGF-β_1 was determined by enzyme-linked immunosorbent assay (ELISA). Immunohistological studies were performed with the streptavidin-peroxidase conjugated methods to assess the expression of TGF-β_1, p-smad3 and Smad7 in sacroiliac joint tissue sample.One-way ANOVA, two independent samples t test and kolmogoorov-Simimov test were used to do statistical analysis. Results In 53 cases patients with AS, 20 cases were with high level Erythro-cyte sedimentation rate(ESR) and C-reactive protein (CRP), while those of the other 33 cases were normal. Serum average TGF-β_1level [ (15.9±5.6) ng/ml ], in patients with high level ESR/CRP [(5.4±5.8) ng/ml ] was significantly increased as compared to the controls and patients with normal ESR/CRP [(4.1±3.6) ng/ml] (P<0.05). There was no expression of TGF-β_1 could be detected in the pannus and bone marrow in SI joints tissue of 30 cases with AS, while decreased level of smad7 expression was detected. In addition, p-smad3 expression was found in the nuclear. Conclusion TGF-β_1 signaling may play an important role in the inflammatory erosion and cartilage fibrosis of sacrojlitis in AS.

Objective To investigate the expression of connective tissue growth factor(CTGF),coll agen I and collagen Ⅲ in sacroiliac joint(SIJ)of patients with spondyloarthropathy(SpA).Methods Thirty patients with SpA,including 17 patients with grade Ⅱ saeroiliitis and 13 patients with grade Ⅰ sacroiliitis,were performed on CT guided needie biopsy of SIJ.After sacroiliitis were confirmed by staining with hematoxylin and eosin in sacroiliac joint tissue sample,immunohistochemical assay was performed to determine the expression of CTGF,collagen Ⅰ and collagen Ⅲ in sacroiliac ioint tissue.Univariate Chi-square test was used for data comparison between multiple groups and t-test was used for two group data comparison.Results Contrast to healthy controls,CTGF were found upexpressed on the cytoplasm of inflammatory cells in pannus and bone marrow of sacroiliac tissue samples of patients with SpA,while collagen I and collagen Ⅲ were found up-expressed in bone,cartilage and ligament tissue[(57.9±42.4)/HP vs(2.7±2.5)/HP P<0.05,0.298±0.080 vs 0.044±0.024 and 28.254±41.165 vs 0.105±0.054.P<0.05 respectively].Conclusion CTGF,collagen Ⅰ and collagen Ⅲ are up-expressed in SIJ of SpA patients.CTGF may play an important role in articular cartilage fibrosis and ossification of SpA.

PURPOSE: Allergic rhinitis (AR) is an inflammatory disorder of the upper airway. Exosomes or extracellular vesicles are nanosized vesicles of endosomal origin released from inflammatory and epithelial cells that have been implicated in allergic diseases. In this study, we characterized the microRNA (miRNA) content of exosomes in AR. METHODS: Extracellular vesicles were isolated from nasal mucus from healthy control subjects (n=10) and patients with severe AR (n=10). Vesicle RNA was analyzed by using a TaqMan microRNA assays Human Panel-Early Access kit (Applied Biosystems, Foster City, CA, USA) containing probes for 366 human miRNAs, and selected findings were validated with quantitative RT-PCR. Target prediction and pathway analysis for the differentially expressed miRNAs were performed using DIANA-mirPath. RESULTS: Twenty-one vesicle miRNAs were up-regulated and 14 miRNAs were under-regulated significantly (P<0.05) in nasal mucus from AR patients when compared to healthy controls. Bioinformatic analysis by DIANA-mirPath demonstrated that 32 KEGG biological processes were significantly enriched (P<0.05, FDR corrected) among differentially expressed vesicle miRNA signatures. Among them, the B-cell receptor signaling pathway (P=3.709E-09), the natural killer cell-mediated cytotoxicity (P=8.466E-05), the T-cell receptor signaling pathway (P=0.00075), the RIG-I-like receptor signaling pathway (P=0.00127), the Wnt signaling pathway (P=0.00130), endocytosis (P=0.00440), and salivary secretion (P=0.04660) were the most prominent pathways enriched in quantiles with differential vesicle miRNA patterns. Furthermore, miR-30-5p, miR-199b-3p, miR-874, miR-28-3p, miR-203, and miR-875-5p, involved in B-cell receptor and salivary secretion signaling pathways, were selected for validation using independent samples from 44 AR patients and 20 healthy controls. MiR-30-5p and miR-199b-3p were significantly increased in extracellular vesicles from nasal mucus when compared to healthy controls, while miR-874 and miR-28-3p were significantly down-regulated. In addition, miRNA-203 was significantly increased in AR patients, while miRNA-875-5p was found to be significantly decreased in AR patients. CONCLUSIONS: This study demonstrated that vesicle miRNA may be a regulator for the development of AR.
B-Lymphocytes ; Biological Processes ; Endocytosis ; Epithelial Cells ; Exosomes ; Humans ; MicroRNAs* ; Mucus* ; Receptors, Antigen, T-Cell ; Rhinitis* ; RNA ; Wnt Signaling Pathway

Objective To establish a non-venous bypass orthotopic liver transplantation model in Bama miniature pigs with high repeatability and stability. Methods Twelve Bama miniature pigs were randomly divided into the donor group (n=6) and recipient group (n=6). Pigs underwent non-venous bypass orthotopic liver transplantation. The time of anhepatic phase during operation was shortened, blood pressure during anhepatic phase was stably maintained, and management of anesthesia and body fluid during operation were strengthened. The operation time, anhepatic phase and survival status of the recipients were observed and recorded. The intraoperative heart rate, mean arterial pressure (MAP) and changes in arterial blood gas analysis were monitored. The perioperative liver function was evaluated. Results Among 6 Bama miniature pigs, 1 died from transplantation failure intraoperatively. The operation time of the remaining 5 pigs was (247±27) min and the time of anhepatic phase was (46±4) min. Three animals survived for more than 2 weeks. Compared with the preanhepatic phase, the heart rate of the animals was significantly faster, MAP was considerably reduced to (46±6) mmHg, blood pH value, base excess (BE) and HCO₃^- level were all significantly decreased and serum level of K⁺ was significantly elevated during the anhepatic phase (all P < 0.05). In the neohepatic phase, MAP of Bama miniature pigs was significantly increased, heart rate was dramatically slower.Blood pH value, BE, HCO₃^- level were significantly increased and serum level of K⁺ was significantly declined (all P < 0.05). During abdominal closure, MAP, blood gas indexes and serum level of K⁺ were almost recovered to those in the preanhepatic phase. Compared with preoperative levels, the levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), lactate dehydrogenase(LDH)and alkaline phosphatase(ALP)were significantly increased after operation (all P < 0.05), the change in AST was the most obvious, and it gradually decreased at postoperative 2 d. The level of γ-gutamyl transferase(GGT) did not significantly elevated. The level of total bilirubin (TB) was evidently elevated at postoperative 5 d. Compared with the preoperative levels, the levels of total protein (TP) and albumin (ALB) were significantly decreased after operation (both P < 0.05), and began to gradually increase at postoperative 1 d. Conclusions The non-venous bypass orthotopic liver transplantation model of Bama miniature pig is convenient, with highly reproducible and survival rate, which can be utilized as a standardized liver transplantation model.