Twenty dyes which previously have been claimed to be excreted in pancreatic juice were reinvestigated to determine to what extent they could be eliminated through the pancreas. Exogenous secretin or cholecysto-kinin-pancreozymin(CCK-PZ) stimuli were used in dogs which had been given intravenous dye solutions at the rate of 1mg/min. In this experiment among the twenty dyes, only six were found to be eliminated through the pancreas. The intensity of dye color in pancreatic juice was estimated photometrically or macroscopically. The dye color intensity decreased as follows; basic fuchsin, acridine red, new fuchsin, rhodamin B, phenol red and rhodamin 6G. Basic fuchsin consistently appeared in CCK-PZ stimulated juice. However, it was seen in only a scant amount or not at all in juice stimulated by purified Vitrum (Sweden) secretin. Similar findings were observed in cats and conscious pigs. The content of basic fuchsin in pancreatic juice was more related to changes in the enzyme concentration than to other components. The chloride content of the juice was related to the amylase or basic fuchsin secretion. However, the chloride content was inversely related to the secreted volume. Vagal stimulation or the administration of parasympathomimetics produced a juice rich in enzyme content, but the dye response to vagal stimulation was weak. Usually the volume of secreted pancreatic juice following stimulation by Boots (England) secretin is greater than stimulated by purified Vitrum preparation. Basic fuchsin was slightly reduced during its elimination from pancreas or when present in alkaline pancreatic juice. Adding acid and formaldehyde revived the color. The acridine red and other pyronine dyes caused the juice to fluorescence. This effect lasted over 24 hours.
Animal ; Dogs ; Dyes/metabolism* ; Pancreas/metabolism* ; Pancreatic Juice/secretion* ; Rosaniline Dyes/metabolism*

OBJECTIVESTo evaluate the application of Coomassie brilliant blue (CBB) G250 staining for the detection of human sperm deformity rate, rate of intact acrosome and acrosome reaction.

METHODSThe smear of spermatozoa before and after capacitation and induced acrosome reaction with progesterone (P) were stained with 0.05% CBB G250 and Wright-Giemisa solution respectively, and visualized with light microscopy. The deformity rate of spermatozoa, rate of intact acrosome and acrosome reaction were calculated.

RESULTSThere was no any difference in detection of deformity rate of spermatozoa and rate of intact acrosome with CBB G250 and Wright-Giemisa staining(P < 0.05). The sperm population of acrosome reaction with induced P was divided by CBB staining into two types: positive staining with dark violet blue on acrosome cap and pale or negative staining on the same area. The rate of the latter was increasing with increasing inductive time, maybe representative of the rate of acrosome reaction. The mean rate was(75.1 +/- 3.8)% after induced for 1 h.

CONCLUSIONSCBB G250 staining is a reliable method for assessment of the human sperm morphology and acrosome reaction.

Acrosome ; metabolism ; Acrosome Reaction ; Humans ; Male ; Rosaniline Dyes ; chemistry ; metabolism ; Spermatozoa ; cytology ; enzymology ; Staining and Labeling

OBJECTIVE: To investigate the relationship between postmortem interval (PMI) and the metabolic law of the amount of DNA in cells. METHODS: After different PMI from the heart, liver, spleen and kidney were taken into pieces respectively, then centrifuged and digested to get suspending cells fluid. The amount of DNA of rats'viscera were detected by flow cytometry after stained by fluorescence, and also inspect the amount of DNA in different periods to find out the law of its variation. RESULTS: It showed a descendent trend of the amount of DNA in cells after different PMI, especially in spleen. CONCLUSION The amount of DNA of all the viscera grows downwards after death, this might be applyed in forensic estimation of PMI.
Animals ; Cell Nucleus/metabolism* ; DNA/metabolism* ; Flow Cytometry ; Kidney/metabolism* ; Liver/metabolism* ; Postmortem Changes ; Rats ; Rats, Sprague-Dawley ; Rosaniline Dyes ; Spleen/metabolism* ; Time Factors

OBJECTIVE: To study the relationship between degradation of marrow DNA and late postmortem interval (PMI). METHODS: Marrow were left on natural condition for 0,1,3,5,7 day after death respectively, Marrow DNA were detected by using Feulgen staining and computerized image analysis. RESULTS: The content of marrow DNA could be detected till 7 days after death yet. CONCLUSION The degradation of marrow DNA may be used on estimation the late PMI.
Bone Marrow/metabolism* ; Cell Nucleus/metabolism* ; DNA/metabolism* ; Humans ; Image Processing, Computer-Assisted ; In Vitro Techniques ; Postmortem Changes ; Rosaniline Dyes ; Sternum/metabolism* ; Time Factors

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

OBJECTIVE: To study the relationship between the DNA content in follicular epithelial cells of the human thyroid and postmortem interval (PMI). METHODS: Changes of the DNA content in thyroid follicular epithelial cells at different PMI were determined by Methyl Green-Pyronin (MGP) stain combined with an image analysis technique. RESULTS: The average DNA content in the thyroid follicular epithelial continued to decrease with increased PMI. The coefficient of determination for DNA content with average gray, aimed area, aimed area ratio and positive index in follicular epithelial cells were 0.960, 0.987, 0.988 and 0.990, respectively. CONCLUSION There is an apparent correlation between the average DNA content of follicular epithelial cells and the PMI. MGP stain combined with an image analysis technique seems to be a useful means to study the degradation of DNA in thyroid follicular epithelial cells.
Adult ; Cadaver ; Cell Nucleus/metabolism* ; DNA/metabolism* ; Epithelium/metabolism* ; Female ; Forensic Pathology ; Humans ; Image Processing, Computer-Assisted/methods* ; Male ; Middle Aged ; Postmortem Changes ; Rosaniline Dyes ; Staining and Labeling/methods* ; Thyroid Gland/metabolism* ; Time Factors ; Young Adult

OBJECTIVETo investigate the protective effect of pSVPoMcat (myelin basic protein microgene) modifying Schwann cell on injured spinal neurons.

METHODSA model of rat spinal cord injured by hemisection was used. One hundred and twenty healthy SD rats of both sexes weighing 250-300 g were divided into three groups: Group A (n=40, treated with implantation of pSVPoMcat modifying Schwann cell), Group B (n= 40, treated with implantation of Schwann cell only) and Group C (n=400, treated with sham operation as the control). One week after operation the rat functional recovery was observed dynamically by using combined behavioral score (CBS) and cortical somatasensory evoked potentials, the spinal cord sections were stained by Nissl, acid phosphatase enzyme histochemistry and cell apoptosis was examined by methye green, terminal deoxynucleotidyl and the dUTP Nick end labeling technique. Quantitative analysis was done by computer image analysis system.

RESULTSIn Group A the injured neurons recovered well morphologically. The imaging analysis showed a result of Group A

CONCLUSIONSpSVPoMcat modifying Schwann cell implantation has protective effect on injured spinal neurons and promotes recovery of injured spinal cord function in rats.

Acid Phosphatase ; metabolism ; Animals ; Apoptosis ; Cell Transplantation ; Disease Models, Animal ; Evoked Potentials, Somatosensory ; Female ; Gene Transfer Techniques ; Male ; Methyl Green ; Myelin Basic Protein ; genetics ; Nerve Regeneration ; Rats ; Rosaniline Dyes ; Schwann Cells ; metabolism ; transplantation ; Spinal Cord Injuries ; physiopathology ; surgery

BACKGROUNDThe mechanism of the neural injury caused by chronic intermittent hypoxia (CIH) that characterizes obstructive sleep apnea syndrome (OSAS) is not clearly known. The purpose of this study was to investigate whether P2X7 receptor (P2X7R) is responsible for the CIH-induced neural injury and the possible pathway it involves.

METHODSEight-week-old male C57BL/6 mice were used. For each exposure time point, eight mice divided in room air (RA) and IH group were assigned to the study of P2X7R expression. Whereas in the 21 days-Brilliant Blue G (BBG, a selective P2X7R antagonist) study, 48 mice were randomly divided into CIH group, BBG-treated CIH group, RA group and BBG-treated RA group. The hippocampus P2X7R expression was determined by Western blotting and real-time polymerase chain reaction (PCR). The spatial learning was analyzed by Morris water maze. The nuclear factor kappa B (NFκB) and NADPH oxidase 2 (NOX2) expressions were analyzed by Western blotting. The expressions of tumor necrosis factor α, interleukin 1β (IL-β), IL-18, and IL-6 were measured by real-time PCR. The malondialdehyde and superoxide dismutase levels were detected by colorimetric method. Cell damage was evaluated by Hematoxylin and Eosin staining and Terminal Transferase dUTP Nick-end Labeling method.

RESULTSThe P2X7R mRNA was elevated and sustained after 3-day IH exposure and the P2X7R protein was elevated and sustained after 7-day IH exposure. In the BBG study, the CIH mice showed severer neuronal cell damage and poorer performance in the behavior test. The increased NFκB and NOX2 expressions along with the inflammation injury and oxidative stress were also observed in the CIH group. BBG alleviated CIH-induced neural injury and consequent functional deficits.

CONCLUSIONSThe P2X7R antagonism attenuates the CIH-induced neuroinflammation, oxidative stress, and spatial deficits, demonstrating that the P2X7R is an important therapeutic target in the cognition deficits accompanied OSAS.

Animals ; Disease Models, Animal ; Hypoxia ; Male ; Metabolic Networks and Pathways ; Mice ; Mice, Inbred C57BL ; Purinergic P2 Receptor Antagonists ; pharmacology ; Receptors, Purinergic P2X7 ; analysis ; physiology ; Rosaniline Dyes ; pharmacology ; Sleep Apnea, Obstructive ; metabolism

Estimation of postmortem interval (PMI) is one of the difficult problems in forensic medicine. With the development of molecular biological techniques, DNA quantification methods were widely applied in estimating PMI. The postmortem degradation of DNA in different tissues and organs was discussed in this article and the recent DNA quantitative techniques being used for estimating PMI were reviewed. These techniques included single cell gel electrophoresis, Feulgen staining image analysis, flow cytometry.
Animals ; Cell Nucleus/metabolism* ; Comet Assay/methods* ; DNA/analysis* ; DNA Degradation, Necrotic ; Flow Cytometry ; Forensic Pathology ; Humans ; Image Processing, Computer-Assisted ; Kidney/pathology* ; Postmortem Changes ; Rosaniline Dyes ; Spleen/pathology* ; Temperature ; Time Factors

The purpose of this study was to investigate the feasibility of sentinel node frozen biopsy to minimize the extensive pelvic lymph nodes dissection in early stage cervical cancer patients on the basis that the risk of skip metastasis to the paraaortic area is negligible. Twenty-six patients with early stage cervical cancer were enrolled in this study. Technetium-99m colloid albumin (Tc(99m)) was injected intradermally around the tumor for allowing preoperative lymphoscintigraphy and intraoperative hand-held gama probe detection of seninel nodes. For visual detection, isosulfan blue dye was injected into the peritumoral sites before peritoneal opening. Postoperative morbidity and negative predictive value were the endpoints of this study. The 26 patients, ranging in age from 32 to 71 yr, underwent intraoperative sentinel nodes mapping. All the patients underwent complete pelvic lymph nodes dissection including para-aortic nodes. There was one case with positive non-sentinel nodes despite the negative sentinel node by frozen biopsy (negative predictive value, 95.2%). This new technique of sentinel node mapping is safe and simple to perform. Further clinical trials using the combination of Tc(99m) and isosulfan blue dye are warranted and this technique will make a true advance for less aggressive management of patients with early stage cervical cancer.
Adult ; Aged ; Female ; Humans ; *Lymph Node Excision ; Lymph Nodes/pathology/radionuclide imaging/*surgery ; Middle Aged ; Pelvic Neoplasms/*pathology/radionuclide imaging/surgery ; Pelvis ; Predictive Value of Tests ; Rosaniline Dyes/metabolism ; Sensitivity and Specificity ; *Sentinel Lymph Node Biopsy ; Technetium Tc 99m Aggregated Albumin/diagnostic use ; Uterine Cervical Neoplasms/*pathology/radionuclide imaging/surgery