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

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

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

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

OBJECTIVE: To observed the pathological changes of closed diffuse brain injury in the rats died immediately and 15 min to 5 days after the injury. METHODS: H.E. staind and esterification-silver stain were applied to investigate the closed diffuse brain injury. RESULTS: In rats died immediately after the concussive injury, a number of shrunken neurons(type I change), distended neurons(type II change) and wave-like nerve fibers were identified in the brain tissue, especially in brain-stem. At 2 h and 8 h after injury, brain edema and axonal swelling appeared clearly in the cortex and white matter, especially in brain-stem. At post-traumatic 8 h and 24 h, the axonal retraction balls began to appear. The amount of neurons undergoing type I and II changes and constraction balls increased along with the survivor time. After 4 days and 5 days, brain edema alleviated, but the retraction balls and axonal swelling still existed. With Esterifica-tion silver stain, the above changes of neurons and nerve fibers were more obvious. With ABC stain, the distribution of albumin(Al) was extended from the perio-vascular area to diffuse distribution. Al positive staining were more obvious in injuried neurons and nerve fibers. CONCLUSION The distribution of the concussive damage in the brain are coup, contra-coup and centripental.
Albumins ; Animals ; Biomechanical Phenomena ; Brain/pathology* ; Head Injuries, Closed/pathology* ; Immunohistochemistry/methods* ; Male ; Rats ; Rats, Sprague-Dawley ; Silver Staining

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

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

This study detects and defines the patterns of p53 gene mutations in breast cancers. We analyse p53 gene mutations through comparing the results of single-strand-conformation-polymorphism (SSCP) and immunohistochemistry (IHC), and we try to define the differences between the results of SSCP and IHC. Twenty-seven fresh primary breast cancer tissues and eight normal breast tissues were studied. The IHC was done with the usual streptavidin-biotin peroxidase complement method by using monoclonal antibody DO-7. The results of staining was scored. The SSCP method was done by using Cold SSCP Electrophoresis System. Overexpressions of p53 protein were seven (25.9%) among 27 cancer cases on IHC. Four (57.1%) of seven cases were positive in SSCP. In SSCP, the mutations were detected in 10 (37%) among 27 cancer cases. The mutations were two in exon 5, one in exon 8, and seven cases in exon 7. All of 10 mutations were proved by sequencing analysis. Of them, only four (40%) were positive in IHC. We consider the IHC as a screening method for p53 gene mutations.
Adult ; Breast Neoplasms/pathology ; Breast Neoplasms/metabolism ; Breast Neoplasms/genetics* ; Comparative Study ; Female ; Gene Expression ; Genes, p53/genetics* ; Human ; Immunohistochemistry ; Middle Age ; Mutation* ; Polymerase Chain Reaction/methods* ; Polymorphism, Single-Stranded Conformational ; Protein p53/genetics ; Protein p53/biosynthesis ; Silver Staining/methods*

Various in situ hybridization (ISH) methods have been used to identify Helicobacter pylori, a causative organism responsible for chronic gastritis and peptic ulcer disease, but they were hard to perform and time consuming. To detect H. pylori in a rapid and easily reproducible way, we developed synthetic biotinylated oligonucleotide probes which complement rRNA of H. pylori. Formalin-fixed and paraffin-embedded tissues from 50 gastric biopsy specimens were examined. Using a serologic test and histochemical stain (Warthin-Starry silver stain and/or Giemsa stain) as a standard, 40 of them were confirmed to be H. pylori-positive. Our ISH was quickly carried out within one hr and results were compared with those obtained from immunohistochemical stain. The ISH produced a positive reaction in 38 of 40 cases (95%). All H. pylori-negative cases failed to demonstrate a positive signal. The ISH has a sensitivity comparable to those of conventional histochemical and immunohistochemical stain, and has high specificity. In conclusion, ISH with a biotinylated oligonucleotide probe provides a useful diagnostic method for detecting H. pylori effectively in routinely processed tissue sections.
Helicobacter Infections/pathology ; Helicobacter Infections/microbiology* ; Helicobacter pylori/isolation & purification* ; Helicobacter pylori/genetics ; Human ; In Situ Hybridization/methods* ; Oligonucleotide Probes ; RNA, Bacterial/analysis ; RNA, Ribosomal, 16S/analysis ; Sensitivity and Specificity ; Silver Staining/methods ; Time Factors