BACKGROUND: The preservation of optimized DNA and its extraction from formalin-fixed, paraffin-embedded (FFPE) tissues are important issues. There has been some doubt over whether 10% neutral-buffered formalin is an ideal fixation solution for DNA preservation over non-buffered formalin, as conventionally recommended. In this study, the correlation between the efficiency of DNA extraction from FFPE tissues and buffered formalin was evaluated. METHODS: Several tissues with same conditions except fixatives were fixed in four different formalin solution groups and were routinely processed as paraffin-embedding protocols. DNAs were extracted from four different FFPE tissues that were stored for over 3 months and over 9 months. The quantity and quality of the DNAs were assessed with a NanoDrop ND-1000 spectrophotometer, and the polymerase chain reaction (PCR) amplification and degradation were analyzed via microchip electrophoresis. KRAS mutation analysis and microsatellite instability (BAT25) PCR were performed with each sample. RESULTS: The results showed no remarkable difference in the four groups. CONCLUSIONS: The study findings demonstrate that DNA preservation is fairly unaffected by a neutral buffer where there is short formalin manufacture period and an adequate formalin fixation time before embedding in paraffin.
DNA ; Electrophoresis, Microchip ; Fixatives ; Formaldehyde ; Microsatellite Instability ; Paraffin ; Pathology, Molecular ; Polymerase Chain Reaction ; Tissue Fixation ; Tissue Preservation

Antineoplastic Agents ; chemistry ; Capillary Electrochromatography ; Chemistry, Pharmaceutical ; Drug Interactions ; Electrophoresis, Capillary ; methods ; trends ; Electrophoresis, Microchip ; Pharmaceutical Preparations ; metabolism ; Pharmacokinetics ; Platinum ; chemistry ; Stereoisomerism

OBJECTIVETo investigate the expression of secretory leucocyte protease inhibitor (SLPI) in colon cancer and their clinical significance.

METHODSImmunohistochemistry was performed to detect the SLPI expression in colon cancer tissue microarray. The expression of SLPI was scored by two pathologists and was analyzed using Χ(2) test to explore its influence on the pathologic characteristics of colon carcinoma.

RESULTSSLPI was up-regulated in colon cancer tissue compared to normal mucosa. Overexpression of SLPI protein was correlated with differentiation grade (low differentiation: 42.1% vs 57.9%; moderate/well differentiation: 2.3% vs 97.7%, TNM stages(III-IV:29.4% vs 70.6%;I-II:3.1% vs 96.9%), lymph node metastasis (28.6% vs 71.4%) and distant metastasis (84.6% vs 15.4%), but not with patient age or sex.

CONCLUSIONSLPI overexpression correlates with aggressive pathologic characteristics of colon cancer and it may server as prognostic factor of colon cancer patients. Further research will be carried out to verify whether SLPI can become a new target for colon cancer treatment.

Adult ; Aged ; Aged, 80 and over ; Colonic Neoplasms ; metabolism ; pathology ; Electrophoresis, Microchip ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Staging ; Secretory Leukocyte Peptidase Inhibitor ; metabolism

The purpose of the present study was to compare the reliability of microchip electrophoresis and capillary electrophoresis for screening FLT3-ITD gene mutation in acute myeloid leukemia. The FLT3-ITD mutation in the genomic DNA samples from 214 untreated AML patients were separately detected by PCR-microchip electrophoresis and PCR-capillary electrophoresis, then the DNA direct sequencing analysis was carried out. The results from PCR-microchip electrophoresis showed that there were 151 FLT3-ITD mutation negative, 58 FLT3-ITD mutation positive (58/214, 27.1%) and 5 FLT3-ITD mutation doubtful positive (5/214, 2.3%), while the outcomes from PCR-capillary electrophoresis displayed that there were 147 FLT3-ITD mutation negative and 67 FLT3-ITD mutation positive (67/214, 31.3%) without doubtful positive. In the 67 FLT3-ITD mutation positive samples detected by using PCR-capillary electrophoresis, 4 samples were detected as the negative while 5 samples were measured as the doubtful positive by using PCR-microchip electrophoresis. The followed sequencing analysis demonstrated that the above 9 samples were all FLT3-ITD mutation positive, indicating that PCR-capillary electrophoresis was more accurate and sensitive in screening the FLT3-ITD mutation, although statistic analysis showed that there were no significant differences in the detected results between PCR-microchip electrophoresis and PCR-capillary electrophoresis groups (Pearson Chi-squared Test, P > 0.05). It is concluded that both PCR-microchip electrophoresis and PCR-capillary electrophoresis were convenient and fast for screening FLT3-ITD mutation, but the accuracy of PCR-microchip electrophoresis awaits further improvement.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Electrophoresis, Capillary ; Electrophoresis, Microchip ; Female ; Humans ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; Male ; Middle Aged ; Mutation ; Young Adult ; fms-Like Tyrosine Kinase 3 ; genetics

AIMTo investigate the roles of acetylcholine (ACh) receptors in the rapid effects of corticosterone (CORT) on the presympathetic neurons in the rostral ventrolateral medulla (RVLM) of rats, and study the non-genomic mechanism of glucocorticoid (GC) in the integration of sympathetic cardiovascular activity.

METHODSThe effects of microelectrophoresis of CORT on the discharge of the presympathetic neurons in the RVLM were observed by extracellular recording in urethane-anaesthetized rats. The responses of atropine (a blocker for M type of ACh receptor, ATR), d-tubocurarine (a blocker for N1 type of ACh receptor, d-TC) and hexamethonium (a blocker for N2 type of ACh receptor, C6) to the effects of CORT on the presympathetic neurons were investigated respectively.

RESULTSTotally 33 presympathetic neurons in the RVLM were recorded. Among them the firing rate of 25 (76%) presympathetic neurons was increased by microelectrophoresis of CORT. The effects of CORT were also positively correlated with the currents. In the other 8 presympathetic neurons, had was shown no effect after microelectrophoresis of CORT. In 10 presympathetic neurons, which discharge was increased by CORT, microelectrophoresis of ATR decreased the firing rate of these presympathetic neurons (P < 0.05), and did not fully block the excitatory effect induced by CORT. In both 7 and 6 presympathetic neurons, application of d-TC and C6 had no effect on these neurons respectively, and did not fully block the excitatory effect induced by CORT.

CONCLUSIONCORT had rapid excitatory effects on the presympathetic neurons in the RVLM, which effect might be independent on ACh receptors.

Animals ; Cholinergic Antagonists ; pharmacology ; Corticosterone ; pharmacology ; Electrophoresis, Microchip ; Male ; Medulla Oblongata ; drug effects ; physiology ; Neuromuscular Nondepolarizing Agents ; pharmacology ; Neurons ; drug effects ; physiology ; Nicotinic Antagonists ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Cholinergic ; physiology