Regional Distribution of Nitric Oxide Synthase in Human Anorectal Tissue: A Pilot Study on the Potential Role for Nitric Oxide in Haemorrhoids.

OBJECTIVE: To study the distribution of nitric oxide synthase (NOS) isoforms and protein levels in human haemorrhoids and rectal tissue. METHODS: Protein expression of NOS1, NOS2 and NOS3 was compared between haemorrhoids (n=14) and normal rectal submucosa (n=6) using Western blot analysis. The localisation of all NOS isoforms to specific structures was determined by immunohistochemistry. RESULTS: Western blot analysis showed median (interquartile range) protein levels of all NOS isoforms were 1.5-2.4 times higher in haemorrhoids than rectal tissue; 121.4 (55.2-165.5) vs 50.0 (25.5-73.7) for NOS1 (p=0.020), 32.2 (23.8-140.6) vs 14.8 (9.6-34.0) for NOS2 (p=0.109), and 80.1 (62.0-139.5) vs 54.3 (48.7 -61.7) for NOS3 (p=0.015). Immunohistochemistry revealed a different distribution and location of all NOS isoforms in vascular and non-vascular structure of haemorrhoids and rectal tissues. The number of haemorrhoid specimens showing positive immunoreactivity of NOS in the vascular endothelium was significantly higher than that in rectal tissue for NOS1 (11/14 (79%) vs 1/6 (17%); p=0.018) and NOS3 (8/14 (57%) vs 0/6 (0%); p=0.042), but not for NOS2 (6/14 (43%) vs 4/6 (67%); p=0.63). CONCLUSION: Haemorrhoids have significantly higher protein levels of NOS1 and NOS3 than rectal tissue. The vascular endothelium of haemorrhoids also has significantly higher positive immunoreactivity of NOS1 and NOS3 than rectal tissue suggesting that blood vessels in haemorrhoids are exposed to higher NO concentrations than those of rectal tissue. Since haemorrhoids exhibit marked vascular dilatation and present with bleeding or swelling, a reduction in NOS - by applying NOS inhibitors - may potentially improve the symptoms of haemorrhoids.

Herbal haemorrhoidal cream for haemorrhoids.

Although hemorrhoids are one of the most common diseases in the world, the exact etiology underlying the development of hemorrhoids is not clear. Many different ointments are currently used to treat hemorrhoids; however, there is little evidence of the efficacy of these treatments to support their use. The aim of this study was to compare different herbal creams used for the treatment of hemorrhoids. Twenty-eight male Wistar albino rats, 6-8 weeks old and weighing 160-180 g, were used in this study as 1-control, 2-croton oil, 3-croton oil+fig leaves+artichoke leaves+walnut husks and 4-croton oil+fig leaves+artichoke leaves+walnut husks+horse chestnut fruit. After 3 days of croton oil application, rats were treated with 0.1 ml of cream or saline twice a day for 15 days by syringe. Tissue and blood samples were collected for histological, immunohistochemical and biochemical studies. Statistical significance was determined using one-way ANOVA followed by Tukey's multiple comparison tests. Croton oil administration resulted in severe inflammation. The third group showed partial improvement in inflammation; however, the greatest degree of improvement was seen in the fourth group, and some recovered areas were observed. Myeloperoxidase immunoreactivity was found to be decreased in the third and fourth groups compared to the second group. Additionally, biochemical analyses (Myeloperoxidase, Malondyaldehyde, nitrate/nitrite and nitrotyrosine levels and Superoxide Dismutase activity) were in agreement with the histological and immunohistochemical results. In conclusion, croton oil causes inflammation in the anal area and results in hemorrhoids. Treatment with our herbal hemorrhoid creams demonstrated anti-inflammatory and anti-oxidant effects in this model.

Haemorrhoids are related to changes of cell function in mucosa and submucosa.

PURPOSE: Epidemiology and risk factors of haemorrhoidal disease are not well defined. This study tried to evaluate if the appearance of haemorrhoids is related to a disturbed remodelling of the soft tissue of rectal mucosa and submucosa. Therefore, immunohistochemical expression profiles of five parameters as potential mediators in neoangiogenesis (EGFR), in inflammatory cell activity (COX-2), and in cell migration, differentiation, and wound healing (notch-3, c-myc, and beta-Catenin) were analysed (Saed et al., Fertil Steril 83(Suppl 1):1216-1219, 1; Saed et al., Fertil Steril 79:1404-1408, 2; Stojadinovic et al., Am J Pathol 167:59-69, 3). METHODS: Haemorrhoidal tissue specimens were collected from 44 patients. Healthy rectal mucosa was obtained from 16 non-fixed fresh cadavers and served as control. Histological and immunohistochemical markers like EGFR, COX-2, notch-3, c-myc, and beta-Catenin were analysed semi-quantitatively, separately for mucosal and submucosal layer. RESULTS: Significantly increased expressions were found for EGFR, COX-2, and notch-3 in the mucosal and submucosal layer of haemorrhoidal tissue in comparison to normal rectal tissue. CONCLUSIONS: This finding confirms that haemorrhoidal disease may be regarded as a manifestation of a soft tissue disease.

High-molecular-mass alkaline phosphatase as a tumor marker for colorectal cancer: comparison of two test methods.

We measured high-molecular-mass alkaline phosphatase (HiMwALP) in serum samples from patients with colorectal cancer by polyacrylamide gel electrophoresis (PAGE) and by column chromatography on diethylaminoethyl (DEAE)-cellulose. Determination of carcinoembryonic antigen (CEA) by an enzyme immunoassay, a frequently used cancer assessment method, was used for comparison. We studied patients with primary colorectal cancer (n = 72), using others with hemorrhoids (n = 38) for a comparison group. HiMwALP activities twice those of pooled normal sera were adopted as cutoff values. The diagnostic sensitivity of the PAGE method for 72 colorectal cancer patients was 63% vs 36% for the DEAE method and 50% for the CEA method. The diagnostic specificities of the PAGE, DEAE, and CEA methods were 89%, 79%, and 95%, respectively. Using both HiMwALP (PAGE method) and CEA for the detection of primary colorectal cancer increased the sensitivity to 72% but decreased specificity to 87%.

Quantitative determination of high molecular weight alkaline phosphatase in patients with colorectal cancer by polyacrylamide gel electrophoresis.

We present a new method for the quantitative determination of high molecular weight alkaline phosphatase (ALP). This method consists of electrophoretic separation on polyacrylamide, incubation of the ALP isoenzymes with substrate, gel drying and quantitative evaluation by densitometric measurement. This method separates high molecular weight ALP from the other isoenzymes. Precision of the method is calculated by the coefficient of variation ranging from 1.2 to 9.5% for samples with different high molecular weight ALP values. The diagnostic sensitivity for detecting colorectal cancer is 60%.