The Expression Of Neuron-specific Enolase (NSE) And S-100in Constipation Patients With Spastic Colon

Objective: Partial colon spasm were ofen observed in patients withconstipation during impact studies or surgery, and the other parts are normal.This experiment use neuron-specific enolase (NSE) and S-100, the specificmarkers of enteric nervous system, to inspect composition of enteric plexuspresence and whether or not there are changes in patients with constipationwith the colon spasm by immunohistochemical examination. Through theexperiment we can explore possible link between spastic colon and entericnervous system, thereby to provide suggestion for therapy.Methods:1Specimen collection1.1Specimen Selection: Specimen Selection: All objects were diagnosed slowtransit constipation by colonic transit experiment, most of the surgicalapproach surgical are subtotal resection of the colon. By expression of impactstudies or surgery, take19patients with spastic colon as the spasm group,20patients without bowel spasms as non-spasm group.In the spasm group,6patients are male, the other13are female, age range is31-74years, themedian age is63years old; in the non-spasm group,3patients are male,17patients are female, age range is18-66years, the median age is48years old.1.2Collection methods: Cut off full thickness intestinal tissue, the size is2×2cm(remove parts of the intestinal wall under clamp). The drawn parts ofspasm group are spastic bowel and transverse colon, the non-spasm group aresigmoid colon and transverse colon.1.3Specimen process: Wash specimen with PBS buffer then placed it into4%paraformaldehyde to fixed24hours. Fish and trim the specimen, dehydratethrough graded alcohol, transparence by xylene, dipping wax, paraffin-embedded specimens. 2Staining2.1Preparation of paraffin sections: Application Rotary microtome to slice,fish sections by warm water flotation, then dry it.2.2Staining: dewax, hydration, antigen retrieval, incubated with1%methanolhydrogen peroxide, after dropping an anti-serum closed (NSE, S-100) and thesecond antibody, stain with DAB, stain with hematoxylin, dehydration ingraded alcohol, finally seal slice.2.3Microscopy: Observe nerve fibers within the enteric nervous system of thestatus and ganglion cell morphology, count the submucosa and muscularplexus in five unoverlapping low magnification vision in the same slicerandomly, then counte ganglion cell within submucosa and muscular plexusunder higher magnification of identification.2.4Image acquisition: When observe abnormal nerve fibers or ganglion cellstake a photograph.3Data processing:This experiment’s measure data is expressed by(x±s)adopting SPSS19.0statistics software, chose two independent samples t-test,test level α=0.05.Results:1Endoscopic findings:1.1NSE staining: Ganglion cells were visible dark brown uniform size anddistribution in submucosal plexus and myenteric plexus of non-spasm group,the nerve fibers aslo rules distribution; campared with the non-spasm group,both number and volume of the dark brown ganglion in the spasm group werechanged, nerve fiber aslo disorders hyperplasia.1.2S-100staining: ganglion cells of control group are multiple cell-like"blank area" arouded by deeply stained plexus both in submucosal plexus andmyenteric plexus, the ganglion cells size and shape is regular, aslo the nervefiber is; the size, shape and quantity of ganglion cells in spasm group changed,diregular distribution and proliferation of nerve fibers can be seen.2NSE staining results:Ganglion cell number: Compared with non-spasm group, ganglion cells of spasm group were decreased both in submucosal plexus and myentericplexus of bowel segment, the difference was statistically significant (P<0.05),ganglion cell number is not statistically difference(P>0.05)in transversesection; plexus quantity: the difference of plexus number between two groupsof patients in the corresponding parts of the intestine was not statisticallysignificant (P>0.05).3S-100staining results:Ganglion cell number: Compared with non-spasm group, ganglion cellsof spasm group were decreased both in submucosal plexus and myentericplexus of bowel segment, the difference was statistically significant (P<0.05),ganglion cell number is not statistically difference(P>0.05)in transversesection; plexus quantity: two groups of patients in the corresponding parts ofthe intestine was not statistically significant (P>0.05).Conclusions:Colon spasm in patients with constipation related to numeber andripeness of ganglion cell within enteric nervous system, diregular distributionand proliferation of nerve fibers salo contribute to partly. Pathology in spasticbowel segment of the control group is similar with Hirschsprung allied disease,these patients didn′t show colonic dilatation may be associated with mildseverity and (or) the proximal colon is in compensated stage.