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| 1 | Degradation of intestinal mRNA:A matter of treatment显示文摘AIM: To characterize the influence of location, species and treatment upon RNA degradation in tissue samples from the gastrointestinal tract. METHODS: The intestinal samples were stored in different medium for different times under varyingconditions: different species(human and rat), varying temperature(storage on crushed ice or room temperature), time point of dissection of the submucous-mucous layer from the smooth muscle(before or after storage), different rinsing methods(rinsing with Medium, PBS, RNALater or without rinsing at all) and different regions of the gut(proximal and distal small intestine, caecum, colon and rectum). The total RNA from different parts of the gut(rat: proximal and distal small intestine, caecum, colon and rectum, human: colon and rectum) and individual gut layers(muscle and submucosal/mucosal) was extracted. The quality of the RNA was assessed by micro capillary electrophoresis. The RNA quality was expressed by the RNA integrity number which is calculated from the relative height and area of the 18 S and 28 S RNA peaks. From rat distal small intestine q PCR was performed for neuronal and glial markers.RESULTS: RNA obtained from smooth muscle tissue is much longer stable than those from submucosal/mucosal tissue. At RT muscle RNA degrades after one day, on ice it is stable at least three days. Cleaning and separation of gut layers before storage and use of RNALater, maintains the stability of muscle RNA at RT for much longer periods. Different parts of the gut show varying degradation periods. RNA obtained from the submucosal/mucosal layer always showed a much worse amplification rate than RNA from muscle tissue. In general RNA harvested from rat tissue, either smooth muscle layer or submucosal/mucosal layer is much longer stable than RNA from human gut tissue, and RNA obtained from smooth muscle tissue shows an increased stability compared to RNA from submucosal/mucosal tissue. At RT muscle RNA degrades after one day, while the stability on ice lasts at least three days. Cleaning and separation of gut layers before storage and use of RNALater, maintains the stability of muscle RNA at RT for much longer periods. Different parts of the gut show varying degradation periods. The RNA from muscle and submucosal/mucosal tissue of the proximal small intestine degrades much faster than the RNA of distal small intestine, caecum or colonwith rectum. RNA obtained from the submucosal/mucosal layer always showed a much more reduced amplification rate than RNA from muscle tissue [β-Tubulin Ⅲ for muscle quantification cycle(Cp): 22.07 ± 0.25, for β-Tubulin Ⅲ submucosal/mucosal Cp: 27.42 ± 0.19].CONCLUSION: Degradation of intestinal m RNA depends on preparation and storage conditions of the tissue. Cooling, rinsing and separating of intestinal tissue reduce the degradation of m RNA. | Sabine Heumüller-Klug Carsten Sticht Karin Kaiser Elvira Wink Cornelia Hagl Lucas Wessel Karl-Herbert Schafer | 2015 | World Journal of Gastroenterology2015，21，12： | 2 |
| 2 | Structure of amyloid βA4-(1-40)-peptide of Alzheimer's disease显示文摘 | STICHT H BAYER P WILLBOLD D | 1995 | Eur J Biochem1995，233，1： | 1 |
| 3 | The theory behind content-based instruction 显示文摘 | Sticht T | 1997 | Focus on Basics1997，，4： | 1 |
| 4 | Structural rearrangements of H1V - 1 Tat- responsive RNA upon binding of neomycin B 显示文摘 | Faber C Sticht H Schweimer K | 2000 | Bio Chem2000，275，20： | 1 |
| 5 | Bidirectional binding of invariant chain peptides to an MHC elass II molecule 显示文摘 | Gunther S Schlundt A Sticht J | 2010 | Proc Natl Acad Sci U S A2010，107，22： | 1 |
| 6 | Magnetic Polarization of the Pd Spacer and Interlayer Magnetic Couplings in Fe/Pd (001) Superlattices:First Principles Calculations显示文摘 | Stoeffler D Ounadjela K Sticht J | 1994 | Phys Rev B1994，49，1： | 1 |
| 7 | Identification and characterization of a eukaryotically encoded rubredoxin in a cryptomonad alga显示文摘 | WASTL J STICHT H MAIER U G | 2000 | FEBS Lett2000，471，： | 1 |
| 8 | Pharmacokinetics of vasoactive substances administered into the human corpus cavernosum显示文摘 | Peskar B Sticht G | 1994 | J Urol1994，151，： | 1 |
| 9 | Spin-Polarized Band Structure of Magnetically Coupled Multilayers显示文摘 | Frank Herman Juergen Sticht Mark Van Schilfgaarde | 1991 | J Appl Phys1991，69，8： | 1 |
| 10 | Frequency, location and age and sex distribution of various types of gastric polyp 显示文摘 | Stolte M Sticht T Eidt S | 1994 | Endoscopy1994，26，8： | 1 |
| 11 | Study of GaN and InGaN Films Grown by Metallorganic Chemical Vapour Depostiton显示文摘 | Van der Sticht W Moerman I Demeester P | 1997 | J Crystal Growth1997，170，14： | 1 |
| 12 | Data quality assessment method for LCI-data of the Dutch building industry 显示文摘 | Schuurmans A Stichting MRPI | 2003 | International Workshop on LCI-Quality2003，，： | 1 |
| 13 | High survivin expression is associated with favorable outcome in advanced primary oral squamous cell carcinoma after radiation therapy显示文摘 | Freier K Pungs S Sticht C | 2007 | Int J Cancer2007，120，4： | 1 |
| 14 | Structural rearrangements of HIV-1 Tat-responsive RNA upon binding of neomycin B显示文摘 | Faber C Sticht H Schweimer K | 2000 | J Biol Chem2000，275，20： | 1 |
| 15 | Activation of MAP kinase signaling through ERK5 but not ERK1 expression is associated with lymph node metastases in oral squamous cell carcinoma (OSCC) 显示文摘 | Sticht C Freier K Knopfle K | 2008 | Neoplasia2008，10，5： | 1 |
| 16 | Gremlin I,frizzled-related protein,and Dkk-I are key regulators of human articular cartilage homeostasis显示文摘 | Leijten JC Emons J Sticht C | 2012 | Arthritis Rheum2012，64，10： | 1 |
| 17 | miRWalk--database:prediction of possible miRNA binding sites by'walking'the genes of three genomes显示文摘 | Dweep H Sticht C Pandey P | 2001 | J Biomed Inform2001，44，： | 1 |
| 18 | The theory behind content--based instruction显示文摘 | Sticht T | 1997 | FocusonBasics1997，，4： | 1 |
| 19 | miRWalk--database:prediction of possible miRNA binding sites by'walking'the genes of three genomes显示文摘 | Dweep H Sticht C Pandey P | 2011 | J Biomed Inform2011，44，5： | 1 |
| 20 | High survivin expression is asso- ciated with favorable outcome in advanced primary oral squamous cell carcinoma after radiation therapy 显示文摘 | Freier K Pungs S Sticht C | 2007 | Int J Cancer2007，120，4： | 1 |
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