OBJECTIVETo investigate the concordance of dual-color silver enhanced in-situ hybridization (DSISH) and immunohistochemistry (IHC) in the detection of HER2 gene amplification and expression and to evaluate the values of DSISH in detecting HER2 gene status in gastric carcinoma.

METHODSBy using automated DSISH and IHC, HER2 gene status was detected in 230 cases of gastric cancer.

RESULTSAmong the 230 cases of gastric carcinoma tested by DSISH, 43 cases were positive and 187 cases were negative; HER2 gene amplification rate was 18.7% (43/230). The expression of HER2 protein was negative, weakly, moderately and strongly positive in 115, 69, 15 and 31 cases, respectively, by IHC. HER2 protein positive rate was 13.5% (31/230). Of the 43 HER2 gene amplification cases by DSISH, 2, 10, 2 and 29 cases were negative, weakly, moderately and strongly positive by IHC; Of the 187 HER2 negative cases by DSISH, 113, 59, 13 and 2 cases were negative, weakly, moderately and strongly positive by IHC, respectively. The overall concordance of HER2 status in the investigation between IHC and DSIDH was 93.5% (201/215), with a high consistency (Kappa coefficient 0.767, P < 0.01).

CONCLUSIONSDSISH can be applied to detect the HER2 gene status in gastric cancer and it also has a high consistency with the result of IHC. In addition, due to frequent heterogeneous expression of HER2, cases with moderate HER2 protein expression may need further assessment by DSISH.

Adult ; Aged ; Aged, 80 and over ; Esophagogastric Junction ; Female ; Gene Amplification ; Genes, erbB-2 ; Humans ; Immunohistochemistry ; In Situ Hybridization ; methods ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Phosphoproteins ; genetics ; metabolism ; Polyploidy ; Receptor, ErbB-2 ; metabolism ; Silver Staining ; Stomach Neoplasms ; genetics ; metabolism

AIDS-Related Opportunistic Infections ; diagnosis ; microbiology ; Acquired Immunodeficiency Syndrome ; diagnosis ; microbiology ; Bronchoalveolar Lavage Fluid ; microbiology ; Female ; Humans ; Immunohistochemistry ; Male ; Methenamine ; Pneumocystis carinii ; isolation & purification ; Pneumonia, Pneumocystis ; diagnosis ; microbiology ; Polymerase Chain Reaction ; Silver Staining ; methods

OBJECTIVETo analyze the proteomic characteristics of Gan (肝)-stagnancy syndrome (GSS) by seeking the differential protein in blood and tissues of GSS model rats.

METHODSGSS model rats were established by chronic restraint stress, keeping rats in restrain chamber for 6 h every day for 21 successive days. Their blood and liver samples were collected at the end of experiment for differential protein detection with methods of isoelectrofocusing and polyacrylamide SDS-PAGE, silver staining, and scanning. The gel images were analyzed with Imagemaster 2D Elite software, and the excavated differential protein spots were identified with matrix assistant laser resolving TOF mass spectrometry, Western blot, ELISA, and RT-PCR, respectively.

RESULTSA method for isolating the protein in blood serum and tissues by two-dimensional gel electrophoresis was established and optimized. Six serum proteins and three liver proteins that differentially expressed were identified. The down-regulated differential proteins in serum of GSS model rats were serum albumin precursor, beta 1 globin, antibody against muscle acetylcholine receptor, Ig lambda-2 C region, and transthyretin (TTR), and those in liver tissue were aryl sulfotransferase, enoyl-CoA hydratase, and TTR. TTR down-regulation was found in both serum and liver. Preliminary biological information analysis showed that these differential proteins involved in immune, neuroendocrine, nutrition, and substance metabolism.

CONCLUSIONProteomic analysis of differential proteins showed that TTR, aryl sulfotransferase, and enoyl-CoA hydratase expressions are downregulated in the GSS model rats, suggesting that the susceptibility of cancer could be enhanced by chronic stress.

Amino Acid Sequence ; Animals ; Chronic Disease ; Disease Models, Animal ; Electrophoresis, Gel, Two-Dimensional ; Liver ; metabolism ; Male ; Molecular Sequence Data ; Prealbumin ; genetics ; Proteomics ; methods ; Rats ; Rats, Wistar ; Reproducibility of Results ; Restraint, Physical ; Silver Staining ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Stress, Psychological ; complications ; metabolism ; Syndrome ; Transcription, Genetic

Biopsy ; Eosine Yellowish-(YS) ; Gastric Mucosa ; microbiology ; pathology ; Helicobacter Infections ; diagnosis ; pathology ; Helicobacter pylori ; isolation & purification ; Hematoxylin ; Humans ; Silver Staining ; Staining and Labeling ; methods

AIMThe comparison of several methods which were used to isolate cochlear outer hair cells and the observations of morphology were researched.

METHODSThree different separating methods of outer hair cells in cochlea were adopted; the morphology of outer hair cells in cochlea and the morphology of cochlear stretched preparation in Silver Nitrate staining were also investigated.

RESULTSSingle alive OHC in cochlea was disassociated by all methods, with microscope and cochlear stretched preparation's staining. We could also observe appearances and distributions of OHC in cochlea.

CONCLUSIONIt is successful to isolate single alive cochlear OHC, and it will be very important to investigate deeply normal physiological functions and changes of functions and morphology in some pathologic status in cochlear OHCs.

Animals ; Cell Separation ; methods ; Cochlea ; cytology ; physiology ; Female ; Guinea Pigs ; Hair Cells, Auditory, Outer ; cytology ; physiology ; Male ; Silver Staining

OBJECTIVEIn this study, orthogonal design was used to optinize DDRT-PCR amplification system on Pinellia ternata microtubers in vitro in five factors four levels respectively.

METHODP. ternata stems and microtubers in vitro were selected as explants. The effects of five kinds of factors were studied by orthogonal design method including emplate cDNA, Mg2+, dNTPs, primers and Taq DNA polymerase, and in order to establish the optimum DDRT-PCR system of P. ternata microtubers in vitro.

RESULT AND CONCLUSIONA satisfactory DDRT-PCR technique system for P. ternata microtubers in vitro with desirable repeatability and polymorphic bands was established. In a total volume of 20 microL DDRT-PCR system, it contained 10 x buffer, 150 micromol L(-1) dNTPs, 2 micromol L(-1) anchor primer, 1 micromol L(-1) arbitrary primer, 2.5 mmol L(-1) Mg2+, 0.6 U Taq DNA polymerase and 2.5 microg template cDNA. The effect of the five factors was in sequence of Taq DNA polymerase > template cDNA > dNTPs > Mg2+ > Primers. The optimum DDRT-PCR system will provide scientific reference basis for studying effecting character of P. ternata microtubers associated with genes expression.

DNA, Complementary ; genetics ; DNA, Plant ; genetics ; Electrophoresis, Polyacrylamide Gel ; Pinellia ; genetics ; Plant Tubers ; genetics ; Polymerase Chain Reaction ; methods ; Silver Staining ; Taq Polymerase ; genetics

OBJECTIVETo verify whether Warthin-Starry (WS) silver method could detect the air particulate matter (PM)/dust particles (Ps) located within the macrophages in situ.

METHODSThere were 26 autopsy cases that resulted from cerebral hemorrhage (group A), silicosis (group B), and fetal death during pregnancy (group C). Samples were collected separately and serial sections were prepared from the lungs and lymph nodes and stained with hematoxylin and eosin (HE), WS silver, immunohistochemistry of CD68. Furthermore, ultrathin sections were taken from the WS positive serial sections of groups A and B. Ps were observed under a transmission electron microscope (TEM) and the elements of Ps were measured by X-ray spectrum analysis (X-RSA).

RESULTSIn both groups A and B, WS staining was positive for the larger and fine Ps, the so called "dust cells", but HE staining was almost negative for fine Ps. In group C, no larger or fine Ps were found. Immunohistochemical staining of CD68 certified that the "dust cells" containing Ps were macrophages. The results of TEM and X-RSA proved that the structure and elements of Ps belonged to PM indeed.

CONCLUSIONWS staining is a better than HE staining in showing the location of PM within macrophages.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Macrophages ; ultrastructure ; Male ; Microscopy, Electron, Transmission ; Middle Aged ; Silver Staining ; methods

OBJECTIVETo investigate histomorphological feature of silicotic nodules under Warthin-Starry (WS) silver staining and its value in the histopathological examination.

METHODSSix cases with silicosis obtained by autopsy and 21 cases with sarcoidosis were collected (among which 3 cases were obtained by autopsy and 18 cases were obtained by biopsy). The serial sections of those paraffin embedded samples were applied respectively for (1) hematoxylin and eosin (HE) staining, (2) WS staining, (3) streptomyces avidin-peroxidase (SP) immunohistochemical staining for mouse anti-human CD68 monoclonal antibody, (4) observing under transmission electron microscope (TEM), (5) X-ray spectrum chemical element analysis(X-RSA). The emphasis of observation and analysis were the dust particles in silicotic nodules and granulomas cells (dust cells, epithelioid cells and multinucleated giant cells in the granulomas). The dust particles deposit in the granulomas were graded under the HE and WS staining.

RESULTSUnder the HE staining the dust particles deposit degrees were (+++) in cellular silicotic nodules, (+) in the fibrous ones, and (-) in the sarcoid nodules; under the WS staining and the dust particles deposit degrees were (+++) in both silicotic nodules whose dust particles were characteristically black, and (+/++) in sarcoid nodules. The dust particles deposit degrees in silicotic nodules were markedly higher than those in sarcoidosis (P < 0.01). The results of immunohistochemical staining indicated that the expression of CD68 in both cells of silicotic nodules and sarcoid nodules were positive. The positive degrees decreased successively with the content of the dust particles. The dust particles of silicotic nodules could be more readily observed than those of sarcoidosis in size and electronic density under TEM. The results of X-RSA indicated that the main chemical element in both dust particles was silicon.

CONCLUSIONWS staining is better than HE staining in showing the dust particles of silicotic nodules, which appear characteristically black, especially in the fibrous ones. Together the TEM observation and X-RSA, the silicotic nodules may be prompted.

Aged ; Aged, 80 and over ; Female ; Humans ; Lung ; pathology ; Male ; Middle Aged ; Silicosis ; pathology ; Silver Staining ; methods

The aim of this study was to compare and analyze protein staining in order to select the optimal staining method for proteomic research. Proteins from acute promyelocytic leukemia cell line NB4 and protein molecular weight marker were separated respectively by 2-D or 1-D electrophoresis and detected respectively by the typical Coomassie brilliant blue, the colloidal Coomassie brilliant blue, the modified Coomassie brilliant blue and the silver staining protocols. The protein detection sensitivity, compatibility with mass spectrometry (MS) and facility of the four staining protocols were compared. The results indicated that the silver staining exhibited the highest sensitivity and MS showed the lowest compatibility 10% of protein identification rate. The detection sensitivity of the modified Coomassie brilliant blue staining was superior to that of other two Coomassie brilliant blue stainings, close to but lower than the silver staining, however the compatibility with MS was better (protein identification rate about 55%). It is concluded that the protein detection sensitivity of the modified Coomassie brilliant blue staining is high, and its compatibility with MS is better, this modified Coomassie brilliant blue staining is an optimal staining method for proteomic research.
Coloring Agents ; Electrophoresis, Gel, Two-Dimensional ; methods ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; Mass Spectrometry ; methods ; Neoplasm Proteins ; analysis ; Rosaniline Dyes ; Silver Staining ; Staining and Labeling

AIMTo screen swimming-fatigue related genes in mice and lay theoretic basis for researching the molecular mechanism of fatigue.

METHODS30 male BALB/c mice (20 +/- 2g) were divided into control group, dipping in water group and swimming-fatigue group respectively. After fatigue for swimming in swimming-fatigue group, with control group and dipping in water group, liver tissues in mice were collected. With improved silver staining mRNA differential display method, the differentially expressed genes in mice livers were screened and evaluated by reversed Northern blot. The positive segments were analyzed homology by BLAST.

RESULTS7 of DD-ESTs were gained. Two of them only expressed in swimming-fatigue group, two down-regulated expressed, and three up-regulated. One of them was a novel gene and was accepted by GenBank, AY615302.

CONCLUSIONSeven DD-ESTs in swimming-fatigue mice were gained by silver staining mRNA differential display method.

Animals ; Fatigue ; metabolism ; Gene Expression Profiling ; methods ; Liver ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; RNA, Messenger ; genetics ; Silver Staining ; Swimming