OBJECTIVETo identify the putative specific localization signal sequence of tumor metastasis suppressor gene-1 (TMSG-1) and to explore the mechanism of subcellular localization of TMSG-1 protein.

METHODSVectors expressing green fluorescence protein (GFP) tagged different TMSG-1 fragments were generated and transfected into human embryo kidney 293 (HEK293) cells. The expression of those fusion proteins was detected by Western blotting and their subcellular localizations were observed by laser confocal microscope.

RESULTSGFP was fused with the native TMSG-1(aa1-380) or different fragments including T1 (aa1-70), T2 (aa1-128), T3 (aa129-380), T4 (aa71-128), T5 (aa71-179) and T6 (aa71-380). Anti-GFP Western blotting showed that these fusion proteins were successfully expressed. Under laser confocal microscope, GFP fused with fragment T4 (aa71-128) localized mainly in the nucleolus; GFP fused with fragment T6 (aa71-380) localized diffusely in the nucleus; while other fusion proteins with TMSG-1 (aa1-380) or fragment T1 (aa1-70), T2 (aa1-128), T3 (aa129-380) and T5 (aa71-179) localized in the cytoplasm. Fragment T4(Δ119-128) was generated from T4 with deletion of 10 amino acid of the C terminal. GFP fused with fragment T4(Δ119-128) remained in the nucleus, but no longer in the nucleolus.

CONCLUSIONSThere is a nucleolar localization signal (aa119-128 RRRRNQDRPS) within TMSG-1. This finding may have laid the foundation for further investigations into subcellular localization and function of TMSG-1.

Amino Acid Sequence ; Blotting, Western ; Cell Nucleolus ; metabolism ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Green Fluorescent Proteins ; metabolism ; HEK293 Cells ; Humans ; Membrane Proteins ; genetics ; metabolism ; Microscopy, Confocal ; Nuclear Localization Signals ; Plasmids ; Recombinant Fusion Proteins ; metabolism ; Sphingosine N-Acyltransferase ; genetics ; metabolism ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVETo investigate the regulatory mechanism of the transcription of tumor metastasis suppressor gene TMSG-1.

METHODSLuciferase reporter assay and site-directed mutagenesis were used to analyze the regulatory region of TMSG-1. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) were carried out to verify the interaction of KLF6 and Sp1 with the regulatory region of TMSG-1. Co-immunoprecipitation (CoIP) was performed to analyze the interaction between KLF6 and Sp1. TMSG-1 and wt-KLF6 mRNA expressions in cells with different metastatic capacities were quantitated by real-time PCR. Cell invasive capability was determined by Matrigel invasion assay.

RESULTSA 63 bp inducible regulatory region (+59 bp - +123 bp) in exon 1 was identified by luciferase assay using reporter plasmids with a series of TMSG-1 regulatory region deletions. Mutations in KLF6/Sp1 binding sites of this region resulted in a decrease of luciferase activity, while cotransfection with KLF6 or Sp1 expressing plasmids led to a remarkable increase of luciferase activity. EMSA and ChIP demonstrated that KLF6 as well as Sp1 interacted with this region. CoIP also indicated a possible interaction between KLF6 and Sp1 proteins. In the highly metastatic cell sublines, a low level of wild type KLF6 was associated synchronously with a low TMSG-1 level. Prostate carcinoma cells overexpressing KLF6 exhibited a higher TMSG-1 level and a lower invasive capability.

CONCLUSIONSTranscription factor complex of KLF6 and Sp1 may participate in the inducible transcriptional regulation of TMSG-1, and a decreased wild type KLF6 expression is likely associated with a low TMSG-1 level in the highly metastatic cell sublines.

Binding Sites ; genetics ; Cell Line, Tumor ; Electrophoretic Mobility Shift Assay ; Humans ; Immunoprecipitation ; Kruppel-Like Factor 6 ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Male ; Membrane Proteins ; genetics ; metabolism ; Mutagenesis, Site-Directed ; Mutation ; Neoplasm Invasiveness ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Sp1 Transcription Factor ; genetics ; metabolism ; Sphingosine N-Acyltransferase ; genetics ; metabolism ; Transcriptional Activation ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVEThe purpose of this review is to discuss some critical issues of isoflavones protective against the development of prostate cancer (PCa).

DATA SOURCESData cited in this review were obtained primarily from PubMed and Embase from 1975 to 2015.

STUDY SELECTIONArticles were selected with the search terms "isoflavone", "Phytoestrogen", "soy", "genistin", and "PCa ".

RESULTSIsoflavones do not play an important role on prostate-specific antigen levels reduction in PCa patients or healthy men. The effect of isoflavones on sex hormone levels and PCa risk may be determined by equol converting bacteria in the intestine, specific polymorphic variation and concentrations of isoflavones. The intake of various types of phytoestrogens with lower concentrations in the daily diet may produce synergistic effects against PCa. Moreover, prostate tissue may concentrate isoflavones to potentially anti-carcinogenic levels. In addition, it is noteworthy that isoflavones may act as an agonist in PCa.

CONCLUSIONSIsoflavones play a protective role against the development of PCa. However, careful consideration should be given when isoflavones are used in the prevention and treatment of PCa.

Humans ; Isoflavones ; therapeutic use ; Male ; Phytoestrogens ; therapeutic use ; Prostatic Neoplasms ; prevention & control

Two Rotavirus G9P[8] strains (LL52696 and LL52727) were recognized during a sentinel-based survey in Lulong, China. Phylogenetic analysis of the VP7 gene showed that both strains isolated constituted a divergent genetic cluster distinct from the other G9 strains isolated in China. Analysis of VP4, VP6, and NSP4 genes revealed that these strains were closely related to Lulong strains. We hold that two strains were reassortant between G9 and Lulong predominant strains.
Amino Acid Sequence ; Antigens, Viral ; chemistry ; genetics ; Base Sequence ; Capsid Proteins ; chemistry ; genetics ; Glycoproteins ; chemistry ; genetics ; Humans ; Phylogeny ; Rotavirus ; classification ; genetics ; Toxins, Biological ; chemistry ; genetics ; Viral Nonstructural Proteins ; chemistry ; genetics

OBJECTIVETo investigate the clinicopathologic features of primary nodal marginal zone B-cell lymphoma (NMZL).

METHODSHematoxylin-Eosin staining and immunohistochemistry were used to evaluate the histological and immunophenotypic characteristics of lymph node (LN) tissue in 22 NMZL cases. Additionally, interphase fluorescence in-situ hybridization (FISH) was carried out to detect the presence of t(11;18) (q21;q21)/API2-MALT1 and/or t(14;18)(q32;q21)/IGH-MALT1 in 9 cases.

RESULTSThe median age of the 22 patients was 62 (16 - 77) ys. The male-to-female ratio was 1.2:1. All patients exhibited asymptomatic lymphadenopathy with the cervical region as the most often site to be involved (n = 11), followed by axillary (n = 9), inguinal (n = 7), submandibular (n = 6), mediastinal (n = 4), supraclavicular (n = 2) and retroperitoneal lymph nodes (n = 1). The Ann Arbor stages were I/II in 13 (59%) cases and III/IV in 9 (41%). Immunohistochemical study showed a consistently strong expression of CD20 and an absence in the expression of CD3ε, CD10, CD21, CD23, CyclinD1 and BCL6 by the tumor cells in all the cases. Frequency of expression of CD5 and BCL2 were 39% (7/18) and 30% (3/14) respectively. Among the 9 cases performed with FISH, 2 cases harbored t(14;18)and another 1 case positive for t(11;18) and t(14;18). Complete follow-up data were available for 13 cases. The follow-up time was 6 to 44 months. 3 of them died. 3-year cumulative survival rate was 67%.

CONCLUSIONSNMZL patients are often elderly, which mainly present with multiple lymphadenopathy, rare involvement of extranodal organ and early stage. The diagnosis must be based on a combination of clinicopathologic features, especially those patients detected t(11;18) and/or t(14;18).

Adolescent ; Adult ; Aged ; Biopsy ; Female ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lymph Nodes ; pathology ; Lymphoma, B-Cell, Marginal Zone ; diagnosis ; pathology ; Male ; Middle Aged ; Prognosis ; Survival Rate ; Young Adult

Objective Lipid metabolism disorders caused by cell foam plays an important role in atherosclerosis,but wheth-er it is involved in the development and progression of silicosis has not yet been elucidated. This study aimed to investigate the effect of free silica(SiO2) in inducing foam cell formation of NR8383 alveolar macrophages in rats. Methods NR8383 cells were cultured in vitro by the routine method (the control group) or in 50 μg/mL SiO2 (the SiO2group), 50 μg/mL ox-LDL (the ox-LDL group), or 50 μg/ml SiO2and ox-LDL (the model group), all for 36 hours. The survival rate of the cells was calculated with the cell proliferation and cytotoxicity assay (MTS),the lipid deposition observed by oil red O staining,the levels of total cholesterol (TC), free cholesterol (FC) and cholesterol esters(CE) measured by ELISA,and the mRNA and protein expressions of PPARγ and CD36 in the cells determined by RT-PCR and Western blot, respectively. Results Compared with the control group,the cells treated with ox-LDL showed a significantly increased survival rate, which reached the peak at 50 μg/mL ([1.501±0.201]%) (P<0.05). Foam cells were observed in the SiO2,ox-LDL and model groups,but most significantly in the model group. In comparison with the ox-LDL group,the model group exhibited remarkable increases in TC([14.195±2.260] vs[35.764±4. 226] μg/mg,P<0.05),FC([7.722±0.690] vs[10.049±0.698] μg/mg,P<0.05),CE([6.473±1.707] vs[25.715±4.243] μg/mg,P<0.05),and CE/TC (45.057% vs 71.642%, P<0.05). Conclusion Free SiO2promotes the lipid metabolism disorder in macrophages and enhances the foaming of the cells,in which PPARγ and CD36 may play an important role of regulation.

;Objective To investigate the influence of pharmacist inter-vention on patients′knowledge of anticoagulation therapy with warfarin . Methods Two hundred sixteen patients who were admitted to cardiovas-cular wards and were prescribed anticoagulation therapy with warfarin from October 2013 to August 2014 were included in the study .According to the ward they stayed , patients were divided into control group ( Cardiology ward No1.) and intervention group ( Cardiology ward No 2.) . In the control group , physicians and nurses introduced anticoagulation knowledge for the patients as usual , while intervention group received medication education and guidance on warfarin use by pharmacists .All patients of two groups filled out an assessment questionnaire about warfa-rin anticoagulation at discharge ( outpoint =13 ) .If any answer to the questions was wrong , pharmacists would educate them again .The score of the questionnaire were compared between the two groups . Results The intervention group enrolled 112 patients while the control group enrolled 104 patients. Scores of assessment questionnaire at discharge of the intervention group and the control group were (10.50 ±2.24) vs (8.08 ±2.61) respectively,with statistical difference ( P <0.05 ) .Conclusion Knowledge of warfarin therapy was much better in patients who received pharmacist intervention than patients who received usual care . Integrated management model with pharmacist interventions can improve patients′cognition in anticoagulation therapy with warfarin .

Objective To detect human parechovirus (HPeV) from stool samples of hospitalized children for acute gastroonteritis of undetectable etiology. Methods We conducted a real-time PCR to detect HPeV. Result The results showed that 24 of 99 (24%) children with gastroenteritis of undetectable etiology were detected with HPeV. Four known HPeV types (HPeV1,3,4,6) were detected in the present study. HPeV1 (50%) was frequently identified as the predominant strain and follow by HPeV3 (25 % ), HPeV 4 (8.3 % ) and HPeV 6 (4.2 % ). We were unable to type 3 samples. Conclusion HPeV was prevalent in hospitalized children for acute gastreenteritis of undetectable etiology in China. Further study is needed for clarifying the role of HPeV in gastroenteritis.

OBJECTIVETo ascertain the causation of a family cluster involving two undefined pneumonia cases, a 12-year-old girl and her brother, reported October, 2005 in Xiangtan county, Hunan province.

METHODSInformation on epidemiology and clinical manifestation of the cases was collected from interviewing the keyman and referring to related medical records. The environment exposure of the cases to their households and the timeline of the illness were reproduced, using this information. Medical check-up was undergone among the close contacts of the cases and on sick/dead poultry. Throat swab of the cases were collected and tested by both RT-PCR and real-time PCR to detect viral nucleic acids of A/H5N1, and were then inoculated into special pathogen free (SPF) embryonated hens' eggs. Serum of the cases including acute and convalescent phases were also collected and tested by microneutralization and haemagglutination-inhibition (HI) assays to detect H5-specific antibodies.

RESULTSBoth the girl and her brother developed fever 2 and 4 days after sudden deaths of chickens being raised in the same house. Both of them had developed pneumonia and the girl died from acute respiratory distress syndrome (ARDS) complicated with multi-organ failure. The boy survived and subsequently discharged from hospital. An eighth-day serum from the girl tested H5 antibody negative, while 4-fold and greater increased in antibody titers were detected in serum from the boy using microneutralization and HI assays in sequential acute and convalescent sera. Of 192 cases, only one doctor who cared for the girl during hospitalization had upper respiratory symptoms but tested negative for H5N1 by microneutralization assay.

CONCLUSIONThe boy was the first confirmed human case of avian influenza A (H5N1) in the mainland of China and his sister was diagnosed clinically. The most probable explanation of these two cases was that the transmission of H5N1 virus from infected poultry within the same household environment. No evidence of human-to-human transmission was noted in the family cluster.

Animals ; Chickens ; Child ; China ; Fatal Outcome ; Female ; Humans ; Influenza A Virus, H5N1 Subtype ; immunology ; isolation & purification ; Influenza in Birds ; transmission ; Influenza, Human ; complications ; diagnosis ; transmission ; Male ; Pneumonia ; virology ; Respiratory Distress Syndrome, Adult ; virology

OBJECTIVETo analyze the high risk factors for tumor recurrence in short term after hepatectomy for the patients with primary hepatocellular carcinoma (HCC).

METHODSFive hundreds and two patients with primary HCC underwent hepatectomy were included from January 1997 to December 2008. Among these patients, males were 419 cases and females were 83 cases. The age was 14 to 82 years (average age 54 years). The results of evaluation on 2 months after resection and tumor recurrence and survival were analyzed.

RESULTSAccording to the operative and pathologic findings and the evaluation on 2 months after hepatectomy, the patients with vascular invasion, palliation resection, cutting edge pathologic residual tumor, lymph notes metastasis, serum AFP level continuing higher after resection or(and) positive TACE (tumor dyeing on TACE within 1 month and a deposit of lipiodol on CT scan) were high risk factors (high-risk group, 106 cases, 21.1%), the recurrence-free survival was 22%, 9% and 3% (1, 2 and 5 year) and overall survival was 52%, 25% and 8%. On the non-high risk group patients, the recurrence-free survival was 84%, 67%, 42% and 31% (1, 2, and 5 year) and overall survival was 97%, 85%, 56% and 35%. The bigger tumor, poor differentiation, tumor invading to liver capsule, satellite focus and TNM III-IV stage in high-risk groups were more significantly than that in non-high-risk groups.

CONCLUSIONThe vascular invasion, palliation resection, cutting edge pathologic residual tumor, lymph notes metastasis, serum AFP level continuing higher or (and) positive TACE within 2 months after resection are high risk factors for HCC patients in short term after hepatectomy, which mean tumor remnant.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; mortality ; pathology ; surgery ; Disease-Free Survival ; Female ; Hepatectomy ; Humans ; Liver Neoplasms ; mortality ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; mortality ; pathology ; Prognosis ; Retrospective Studies ; Risk Factors ; Young Adult