Objectives To screen the differentially expressed proteins in serum of population chronically exposed to arsenic in drinking water,thus to provide candidate protein biomarkers for arsenic exposure and arsenicosis.Methods Subjects were selected from the drinking water type of endemic arsenicosis areas in Shanxi province,China.Demographic characteristics,history of arsenic exposure,cigarette smoking,alcohol drinking,health and other information were collected using questionnaire.The subjects were divided into low-arsenic group (with arsenic in drinking water ＜ 10 μg/L),medium-arsenic group( 10 - 50 μg/L),high-arsenic group( ＞ 50 μg/L),and arsenicosis group(the drinking water with arsenic ＞ 50 μg/L was replaced by low arsenic water ＜ 10 μg/L).The number of cases in each group was 30.The arsenicosis patients were diagnosed according to “Standard of Diagnosis for Endemic Arsenism” (WS/T 211-2001 ).With the principle of informed consent,blood samples were collected.Differentially expressed serum proteins of different arsenic exposure groups and arsenicosis group were screened by two-dimensional differential gel electrophoresis(2-D DIGE),and further identified by mass spectrometry (MS).Results An average of (1299 ± 167) protein spots were identified in 6 gel images and 688 protein spots were discovered repeatedly in at least 5 gels.There were 33 protein spots differentially expressed among low-,medium- and high-arsenic groups P ＜ 0.01).Fifty four protein spots were significantly different among low-,medium-,high-arsenic exposure groups and arsenicosis group(P ＜ 0.01 ).Twenty five protein spots were selected for MS analysis,and13 protein spots were identified.Compared with low-arsenic group,the expressions of apolipoprotein A-Ⅳ,retinol binding protein,and estrogen receptor hypothalamic isoform in medium- and higharsenic exposure groups were down regulated,and the expressions of component 4A and 4B were up regulated.Compared with low-,medium- and high-arsenic groups,the expressions of beta-2-glycoprotein Ⅰ,Keratin 1,hemopexin,complement C1r subcomponent,and ficolin-3 in arsenicosis group were down regulated,and the expressions of pigment epithelial-differentiating factor,alpha-1-microglobulin and carboxypeptidase N catalytic chain were up regulated.Conclusions Chronic arsenic exposure can significantly change population's serum protein expression.Differentially expressed proteins in arsenicosis patients will not decline with the decline of arsenic in a short term.Whether or not the differentially expressed proteins identified in this study can be used as biomarkers for arsenic exposure and arsenicosis needs to be further verified.

Objective To evaluate the effect of various iterative reconstruction methods on phase analysis of gated myocardial perfusion imaging (MPI). Methods Thirty consecutive patients scanned by the Philips CardioMD system were recruited into this study. The gated SPECT (GSPECT) data were reconstructed with filtered backprojection (FBP),maximum likelihood expectation maximization (MLEM),three-di-mensional (3D) resolution recovery MLEM (AST),attenuation corrected (AC) MLEM,AC and 3D Monte Carlo scatter corrected (ACSC) MLEM methods. Parameters of left ventricular ( LV ) dyssynchrony ( phase standard deviation and histogram bandwidth) were measured using the software SyncTool. Paired t-test was used to compare the differences of the LV dyssynchrony indices between FBP and MLEM,AC MLEM,ACSC MLEM,AST respectively. Results The phase standard deviations of stress GSPECT MPI for FBP,MLEM,AC MLEM,ACSC MLEM,and AST were 11.6°,10.9°,11.2°,11.6°,11.4° respectively;while the histogram bandwidths were 35.7°,34.3°,35.1°,36.9°,35. 1 ° respectively. The phase standard deviations of rest GSPECT MPI for FBP,MLEM,AC MLEM,ACSC MLEM and AST were 15.2°,14. 5°,15.4° ,15. 4°,14.8° respectively; while the histogram bandwidths were 47.3°,46.4°,46.4° ,47.9°,46.1 ° respectively. No statistical significance was observed between the FBP and various iterative reconstruction methods for both the stress and rest GSPECT MPI study (t:-1. 179 to 1.554,P＞0.05 forall). Conclusion The standard FBP reconstruction method is accurate enough for the measurement of LV dyssynchrony indices using the widely used clinical software SyncTool.

ObjectiveTo observe the changes of fluoride content in serum,bone and urine after rats were exposed to single fluoride,single aluminum or fluoride combined with aluminum and to investigate the effects of different doses of aluminum on fluoride accumulation and excretion in rats.Methods Male Wistar rats were randomly divided into 9 groups based on 3 × 3 factorial design.Different doses of fluoride(NaF,0,50,200 mg/L)and(or) aluminum(AlCl3,0,100,200 mg/L) were administered to rats in each group by drinking water.The rats took food and water ad libitum during the experimental period.After feeding for 18 weeks,rats with obvious dental fluorosis were determined as successful establishment of animal model.The fluoride content in the serum,bones and urine were measured.Results Fluoride affected the fluoride content in serum,bones and urine(F=166.74,577.81,160.96,all P ＜ 0.01 ).The interaction of fluoride and aluminum on serum,bone and urinary fluoride were statistically significant (F =7.95,5.13,6.94,all P ＜ 0.01 ).When the fluoride level was 50 mg/L,the serum fluoride contents were [ (0.08 ± 0.03) and (0.08 ± 0.02) mg/L] in the aluminum levels of 0 and 100 mg/L groups,which was higher than that of the aluminum level of 200 mg/L group[ (0.04 ± 0.01)mg/L,F=7.14,5.78.all P＜ 0.05].The bone fluoride content in the 0 mg/L aluminum level group[ (1996.53 ± 383.73) mg/kg] was higher than that of the 100 and 200 mg/L groups[(1252.51 ± 189.08),( 1160.63 ± 129.63) mg/kg,F=20.54,24.56,all P ＜ 0.01 ].When the fluoride level was 200 mg/L,the bone fluoride contents were decreased with the increasing doses of aluminum[ (4668.70 ± 887.67),(3920.30 ± 528.31 ),(3297.64 ± 396.04) mg/kg].Between any two groups,the differences were statistically significant (F =15.59,52.31,14.38,all P ＜ 0.01 ).When the fluoride level was 50 mg/L,the urinary fluoride content in the 0 mg/L aluminum level group[ (34.054 ± 9.30)mg/L] was higher than that of the 100,200 mg/L groups[( 14.81 ± 6.32),(14.67 ± 3.42) mg/L,F =25.30,24.32,all P ＜ 0.01 ].When the fluoride level was 200 mg/L,the urinary fluoride contents in the 0,100 mg/L aluminum level groups[ (57.14 ± 21.38),(51.75 ± 8.39)mg/L] were higher than that of the 200 mg/L group[(34.839 ± 9.30) mg/L,F=30.04,20.31,all P ＜ 0.01 ].ConclusionsAluminum is an antagonist of fluoride.The antagonism could be enhanced as the dose of aluminum increased.In this study,aluminum could effectively counteract the absorption of fluoride in rat model when the ratio of fluoride to aluminum is 1 ∶ 2.

OBJECTIVESTo investigate the intralaboratory reproducibility of immunohistochemistry (IHC) testing for HER2 status in breast cancer, and to evaluate the factors which influence the reproducibility. The concordance between monoclonal antibody CB11 and HercepTest was also assessed.

METHODSHER2 overexpression on paraffin sections from thirty-seven cases of breast invasive ductal carcinoma was evaluated using CB11 and the evaluation procedure had been repeated for five times scored the tests together according to the HercepTest and new American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) grading schemes by 2 experienced pathologists together. Reproducibility rates of the five rounds were assessed using Kappa statistic, and the results from two scoring systems were compared. HercepTest kit was applied to the same cases afterward and the results were compared with CB11.

RESULTSSubstantial intralaboratory reproducibility was achieved among 5 rounds tests. Excluding the influence effect of changing antibody lots, the intralaboratory reproducibility was closed to the perfect threshold (Kappa = 0.7858, HercepTest scheme). The results derived from the two grading schemes had an almost perfect agreement (Kappa = 0.8549). The concordance (positive vs. negative) between CB11 and HercepTest was 83.78%.

CONCLUSIONSLaboratory work with strict supervision and more experience will ensure a reliable testing consistency. Reproducibility analysis could be adopted to evaluate the intralaboratory staining quality on HER2 testing. Different antibody lots bring some influence to the intralaboratory reproducibility, but not significant. CB11 could be accepted to screen HER2 status in routine practice after testing validation.

Antibodies, Monoclonal ; metabolism ; Breast Neoplasms ; metabolism ; Carcinoma, Ductal, Breast ; metabolism ; Female ; Humans ; Immunohistochemistry ; methods ; Receptor, ErbB-2 ; metabolism ; Reproducibility of Results

OBJECTIVETo study the effect on anti-respiratory syncytial virus of an active compound (AP3) from a Chinese medicinal herb-Herba patriniae in vitro.

METHODSActive component of herba patriniae (AP3) was extracted and its anti-respiratory syncytial virus (RSV) effect was tested. A water soluble substance (AP3) was isolated from a Chinese herb Herba patriniae, by hot water extraction, ethol precipitation and gel permeation column chromatography. The cytotoxicity of AP3 was tested by adding it to HeLa cells directly. Its effect against RSV was estimated by CPEI assay while ribavirin was used as positive control.

RESULTSChemical test showed that the nature of substance AP3 was polysaccharide. The median cytotoxic concentration (TC(50)) of AP3 was 11.45 mg/ml by morphological observation and the median effective concentration (50% effective concentration, EC(50)) of it against replication of the long strain of RSV in HeLa cells was 0.0986 mg/ml. The Therapeutic index (TI = TC(50)/EC(50)) of AP3 was 116.12, much higher than the TI of herba patriniae (AP1) (TI = 59.26) and ribavirin (TI = 53.45). Moreover, AP3 gave a dose-dependent response in inhibiting RSV. In the assay, the effect of AP3 against RSV growth was also tested. In addition, the effect of AP3 on virus growth, AP3 inhibited replication of RSV in HeLa cells, when added at 0 h, 2 h, 4 h after virus infection, were also tested.

CONCLUSIONThis study suggested that the AP3 exerted an obvious inhibitory effect to RSV in HeLa cell culture. This study furnished a reliable evidence for development of a new antiviral drug.

Antiviral Agents ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; HeLa Cells ; Humans ; Respiratory Syncytial Viruses ; drug effects ; Time Factors

OBJECTIVETo explore the prognostic factors of primary nodal diffuse large B-cell lymphomas (N-DLBCL).

METHODSAccording to the 2001 WHO classification of tumors of hematopoietic and lymphoid tissue, 51 cases of primary N-DLBCL were collected for clinical data analysis and immunohistochemical assay. Antibodies used for study were anti-CD20, CD79alpha, CD45RO, CD3, Bcl-2, Ki-67, CD30, CD15, kappa, lambda, Cyclin D1, TdT, GFAP, CK, MPO. The survival data was analyzed.

RESULTSOf the 51 cases of N-DLBCLs, 40 were reclassified as centroblastic, 3 B-immunoblastic, 1 T-cell/histiocytes rich, 2 B-cell anaplastic large cell, 1 plasmablastic, and 4 unclassified. Expression of Bcl-2 oncoprotein was observed in 24 cases (47.1%). The median Ki-67 index was 50.0% and the index more than 40% was found in 35 cases (68.6%). Survival analysis of 35 cases had follow up data showed that the 2 year and 5-year overall survival (OS) rates were 48.54% and 35.30%, respectively. The 5-year OS rates patients with International Prognosis Index (IPI) >/= 3 was lower than that with IPI < 3 (P < 0.01). The 5-year OS rates for patients with B symptoms was lower than that without B symptoms (P < 0.05). The 5-year OS rates for patients with Ki-67 index more than 40% was lower than that with less than 40% (P < 0.05). The expression of Bcl-2 oncoprotein was uncorrelated to prognosis (P > 0.05).

CONCLUSIONIPI, B symptoms and Ki-67 index are the prognostic factors for patients with N-DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD20 ; analysis ; Child ; Child, Preschool ; Cyclin D1 ; analysis ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Ki-1 Antigen ; analysis ; Ki-67 Antigen ; analysis ; Leukocyte Common Antigens ; analysis ; Lewis X Antigen ; analysis ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Male ; Middle Aged ; Prognosis ; Survival Analysis ; Young Adult

OBJECTIVETo investigate the expression of B cell-specific activator protein (BSAP)/Pax-5 in lymphomas.

METHODSOne hundred and two cases of diffuse large B-cell lymphoma (DLBCL), 3 cases of follicular lymphoma (FL), 3 cases of extranodal marginal zone B-cell lymphoma, 1 case of nodular lymphocyte predominant Hodgkin lymphoma (NLPHL), 10 cases of anaplastic large cell lymphoma (ALCL) and 10 cases of plasmacytoma were studied immunohistochemically for BSAP and CD20.

RESULTSThe tumor cells in the 102 cases of DLBCL all expressed CD20, amongst which 100 cases also expressed BSAP. Three cases of FL, 3 cases of extranodal marginal zone B-cell lymphoma and 1 case of NLPHL also expressed BSAP and CD20. All the ALCLs and plasmacytomas did not express BSAP and CD20. The expression rates of CD20 and BSAP were highly consistent. The intensity of staining showed no statistical significance.

CONCLUSIONSBSAP/Pax-5 is a novel B-cell marker expressed in tumor nuclei of B-cell lymphomas. Though less sensitive than CD20, anti-BSAP has diagnostic value in routine surgical pathology practice.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD20 ; metabolism ; Biomarkers, Tumor ; Cell Nucleus ; metabolism ; Child ; Child, Preschool ; Female ; Humans ; Lymphoma, B-Cell ; metabolism ; Lymphoma, Follicular ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; Male ; Middle Aged ; PAX5 Transcription Factor ; metabolism ; Plasmacytoma ; metabolism

Objective To evaluate the accuracy of automated quantification of myocardial perfusion imaging (MPI) using a method based on a Western normal database for the detection of coronary artery disease (CAD) in a group of Chinese patients. Methods Seventy-two Chinese patients who underwent coronary angiography (CAG) and MPI within 3 months were recruited into this study. Eighty selected from 140 Chinese patients with low probability of CAD ( ＜ 5% ) were enrolled into local normal database of 99Tcm-methoxyisobutylisonitrile (MIBI) MPI using Cedars quantitative perfusion SPECT (QPS) database. Two Western MPI normal databases (CSMC MibiMbiAuto and Mibimibi) were used for processing the Chinese CAD patients recruited in this study, and the results were compared with those using local normal database and visual interpretation. T-test and z-test were used for statistical analysis. Results The extent (EXT)measurement obtained from Mibimibi and local database was ( 10.73 ± 14.54)% and ( 14.22 ± 16.51 )%,respectively ( t = 7.87, P ＜ 0.001 ); the severity (SEV) was 1.07 ± 0.93 and 1.34 ± 1.20, respectively ( t =7.45, P＜0.001). The area under curve(AUC) by using EXT measurement for local database (0.85 ±0.05) was larger than that for CSMC MibiMbiAuto ( AUC = 0.72 ± 0.06, z = 2.50, P ＜ 0.01 ) and Mibimibi ( AUC = 0.77 ± 0.06, z = 2.47, P = 0.014). The AUC of local database showed no significant difference from that of visual interpretation (AUC=0.83 ±0.05, z=0.05, P＞0.05). Conclusion Quantification of MPI of our Chinese patients using Western normal database would decrease the accuracy for the detection of CAD.

OBJECTIVETo evaluate pathomorphologic and immunohistochemical characteristics of the bone marrow involvement of lymphoma and its significance in the diagnosis and subtype of lymphoma with bone marrow involvement.

METHODSOne hundred and eighty eight formalin fixed and paraffin embedded bone marrow biopsy specimens were studied. Immunohistochemical staining was performed.

RESULTS(1) Five patterns of bone marrow involvement of lymphoma were found, including diffuse (44.9%), focal (29.3%), interstitial (11.6%) and nodular (6.1%). (2) There were many subtypes of lymphoma in these cases, the most common type was lymphoplasmacytic lymphoma (21.7%). (3) The lymphomas in bone marrow biopsy had their own special characteristics of morphology and immunophenotype as did in extra-medullar lymphomas. (4) Fibrosis (75.8%) and hematopoietic tissue hypoplasia (71.1%) were found in most cases and necrosis in a few cases.

CONCLUSIONSMost cases of bone marrow involvement of lymphoma could be diagnosed and classified by combination of histopathological and immunohistochemical analysis. Diagnosis of some cases could be made only after the review of pathological changes of lymph node. A few cases were difficult to classify their subtypes of lymphoma.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Marrow ; pathology ; Child ; Female ; Humans ; Immunophenotyping ; Lymphoma ; diagnosis ; immunology ; pathology ; Male ; Middle Aged ; Neoplasm Invasiveness

OBJECTIVETo study the effects of human telomerase reverse transcriptase (hTERT) RNA interference (RNAi) on biological characteristics of hepatocellular carcinoma cell lines HepG2 and SMMC-7721 and on apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL).

METHODSSmall hairpin hTERT (shTERT) sequence was identified by PCR method; hTERT expressions, morphological features, cell proliferation and replicative senescence were respectively determined using RT-PCR, hematoxylin-eosin (HE) staining, growth curve and beta-galactosidase (b-Gal) staining; cell cycle and apoptosis were identified using flow cytometry after propidium iodide (PI) staining and annexin V/PI double staining.

RESULTSshRNA were found in 6/8 HepG2 and 6/6 SMMC-7721 cell clones transformed by the recombined plasmid pSilencer 3.1-H1 neo-shTERT. The interference rates of hTERT on HepG2 and SMMC-7721 were 100% and 43.3% respectively. Cells in G2-M phases increased from 7.1% to 10.6% and from 6.9% to 7.9% respectively; and the percentage of replicative senenscence cells increased from 0 to 20.4% and from 3.6% to 10.0% respectively. The nucleus/cytoplasm ratios of the cells were obviously decreased after hTERT RNAi treatment. Moreover, apoptosis of hepatocellular carcinoma cells and apoptosis induced by TRAIL were strikingly increased by hTERT RNAi (P < 0.05). For example, apoptosis rates were increased from 3.5% to 5.2% in HepG2 cells and from 4.8% to 7.9% in SMMC-7721 cells after hTERT RNAi treatment. Apoptosis rates were increased from 5.3% to 10.4% in HepG 2 cells and from 13.9% to 77.2% in SMMC-7721 cells after being treated by 100 ng/ml TRAIL for 24 h. However, there were no remarkable changes between control cells and untransformed cells.

CONCLUSIONhTERT RNAi not only has a significant effect on biological characteristics of hepatocellular carcinoma cells, but also obviously can increase cell apoptosis induced by TRAIL.

Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Humans ; Liver Neoplasms ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Telomerase ; genetics ; Tumor Cells, Cultured