Objective To establish interstitial cells of Cajal （ICC） loss models caused by incomplete small intestinal obstruction in rats with modified method and verify it. Methods Modified method was used to establish the models， making the ring around the small intestine but not through it. Morphological changes were observed by general signs， pathological changes were tested by HE staining and transmission electron microscope （TEM）， and changes of ICC number were tested by immunohistochemistry staining. Results Success rate of this method was 56% （28/50）， weight loss happened compared with before operation in ileus group （P＜0.01）. Hyperemia and swelling were observed in ileus group， and gastric retention was obvious. Results of HE staining and TEM showed that there was obvious inflammatory change， and ICC reduced was observed by immunohistochemistry. Conclusion ICC loss models caused by incomplete small intestinal obstruction meet the basic performance， and can be used for further research.
Abstract: Objective To induce calcification in aortic valvular interstitial cells (VICs) in vitro and observe the shift of cellular phenotype during the process. Methods Porcine aortic VICs were isolated and expanded by collagenase methods. Fluorescent staining was performed to identify the interstitial cells. VICs at 48 passages were used for experiments. The cells were divided into two groups: the experimental group in which cells were cultured in osteogenic media supplemented with βglycerophosphate, vitamin C and dexamethasone, and the control group in which cells were cultured in normal media. After 2 weeks, calcified nodules were quantified. Calcium deposit was stained and measured by Alizarin Red S staining and assay. Real time reverse transcription polymerase chain reaction (RTPCR) was performed to measure expression of alpha smooth muscle actin (α-SMA) and calcification related factors such as osteocalcin, osteopontin and Corebinding factor α1/Runx2 (Cbfα1/Runx2). Results VICs were successfully harvested from porcine aortic valves, identified by positive staining of α-SMA, vimentin and negative staining of Von Willebrand factor (vWF). VICs could calcify after 2 weeks of osteogenic induction with calcified nodules formed. Quantification of calcified nodules and calcification deposit were significantly higher (Plt;0.05) in the experimental group than those in the control group (156.25±17.38 vs. 2.50±1.29, 17.52±2.04 vs. 1.00±0.22). Real Time RT-PCR indicated that expression of α-SMA, as well as calcification related markers like osteocalcin, osteopontin and Cbfα1/Runx2 was much higher in the experimental group than those in the control group (Plt;0.05). Conclusion VICs are activated during the progress of calcification with phenotype shifting to contraction and ossification, which might be the pathological basis of valvular calcification.
Objective To investigate the express of ERβ protein in female slow transit constipation (STC) patients. Methods Immunohistochemistry and Western blot technique were used to detect the distribution and expression of estrogen receptor β (ERβ) protein of 20 patients with STC and 20 aged-matched controls. Results ERβ expressions were detected in mucous layer, myenteric nerve plexus and submucous nerve plexus in two groups. In comparison with the control group, the expression of ERβ protein of STC group was much lower (Plt;0.01). The expression of ERβ protein of sigmoid colon in STC group was significantly lower than that in control group (Plt;0.05). Conclusion The expression of ERβ protein decreased in myenteric and submucous nerve plexus of sigmoid colon tissues may involve in the pathogenesis of STC.
Objective To study the effects of long term application of cathartics on electromyography of rat colon, and to explore the role of interstitial cells of Cajal (ICC) in it. Methods Colonic slow waves of the rat was examined after 3-month feeding of phenolphthalein, and ICC in myenteric plexus was observed by ZIO method, and ultrastructure changes of nerves and ICC was observed. Results The frequency of slow waves of cathartic colon was reduced significantly(Ｐ＜0.05). The distribution of ICC in myenteric plexus was uneven, and the processes were mussily connected each other. Vacuolar degeneration of axon and ICC-like cells was revealed by electron microscope in myenteric plexus of cathartic colon. Conclusion Long term application of phenolphthalein could reduce the frequency of colonic slow waves, and the possible mechanism was degeneration of ICC and myenteric plexus nerves.
ObjectiveTo investigate the changes of amount of interstitial cells of Cajal (ICC) and expression of stem cell factor (SCF)/c-Kit in the process of cathartic colon induced by emodin in mice. MethodsA modified cathartic colon mouse model was established. Seventy-two healthy male Kunming (KM) mice were randomly divided into the blank control group and sustained drug delivery group.Morphological changes of colon in mice were observed; frozen section immunofluorescence was used to observed changes of amount of ICC; serum concentrations of SCF were examined by ELISA; Western blot was employed for observation of expression of SCF/c-Kit in colon. ResultsAfter the mice model were completed, the weight of mouse, length and diameter of entire colon were all reduced compared with the blank control group. The amount of ICC appeared to decline in the beginning of the first 6 weeks with emodin used, and significant decreased in 10-12 weeks. The serum concentrations of SCF first began to decline in 4 weeks with emodin used, and significantly decreased in 6 weeks, and continued at a low level after 8 weeks. The expression of c-Kit in colon began to decline in 4 weeks with emodin used and significantly reduced after 8 weeks. Conciusions The amount of ICC appear to slowly decline in the beginning of the first 12 weeks with emodin used, and significant decrease after 12 weeks.The serum concentrations of SCF and expression of c-Kit in colon have the dynamic changes in the meanwhile, and the changes of SCF are earlier than that of c-Kit. The trend of amount ofICC may have a certain relationship with changes of SCF and c-Kit.