The analysis of boric acid effect on epithelial-mesenchymal transition of CD133 + CD117 + lung cancer stem cells.

Targeting lung cancer stem cells (LC-SCs) for metastasis may be an effective strategy against lung cancer. This study is the first on epithelial-mesenchymal transition (EMT) properties of boric acid (BA) in LC-SCs. LC-SCs were isolated using the magnetic cell sorting (MACS) method. Tumor-sphere formation and flow cytometry confirmed CSC phenotype. The cytotoxic effect of BA was measured by MTT analysis, and the effect of BA on EMT was examined by migration analysis. The expression levels of ZEB1, SNAIL1, ITGA5, CDH1, ITGB1, VIM, COL1A1, and LAMA5 genes were analyzed by RT-qPCR. E-cadherin, Collagen-1, MMP-3, and Vimentin expressions were analyzed immunohistochemically. Boric acid slightly reduced the migration of cancer cells. Increased expression of transcription factor SNAIL (p < 0.001), but not ZEB1, was observed in LC-SCs. mRNA expression levels of ITGB1 (p < 0.01), ITGA5 (p < 0.001), COL1A1 (p < 0.001), and LAMA5 (p < 0.001) increased; CDH1 and VIM decreased in LC-SCs. Moreover, while E-cadherin (p < 0.001) and Collagen-1 (p < 0.01) immunoreactivities significantly increased, MMP-3 (p < 0.001) and Vimentin (p < 0.01) immunoreactivities decreased in BA-treated LC-SCs. To conclude, the current study provided insights into the efficacy and effects of BA against LC-SCs regarding proliferation, EMT, and cell death for future studies.

Comparison of exosomes secreted by synovial fluid-derived mesenchymal stem cells and adipose tissue-derived mesenchymal stem cells in culture for microRNA-127-5p expression during chondrogenesis.

This study aimed to investigate the differences between the exosomal microRNA-127-5p expression profiles of human adipose tissue-derived mesenchymal stem cells (hAT-MSCs) and human synovial fluid-derived mesenchymal stem cells (hSF-MSCs) during chondrogenesis in terms of regenerative treatment of cartilage. Synovial fluid-derived mesenchymal stem cells, adipose tissue-derived mesenchymal stem cells, and human fetal chondroblast cells (hfCCs) were directed to chondrogenic differentiation. Alcian Blue and Safranin O stainings were performed to detect chondrogenic differentiation histochemically. Exosomes derived from chondrogenic differentiated cells and their exosomes were isolated and characterized. microRNA-127-5p expressions were measured by Quantitative reverse transcription PCR (qRT-PCR). Significantly higher levels of microRNA-127-5p expression in differentiated hAT-MSCs exosomes, similar to human fetal chondroblast cells, which are the control group in the chondrogenic differentiation process, were observed. hAT-MSCs are better sources of microRNA-127-5p than hSF-MSCs for stimulating chondrogenesis or in the regenerative therapy of cartilage-related pathologies. hAT-MSCs exosomes are rich sources of microRNA-127-5p and can be an essential candidate for cartilage regeneration treatments.

The potential therapeutic effect of klotho on cell viability in human colorectal adenocarcinoma HT-29 cells.

Klotho is an anti-aging, anti-inflammator, and anti-oxidative protein and has been shown to important role in tumorigenesis, proliferation, survival, autophagy, and resistance to tumor suppressor effects in several types of human cancers. In this study, we aimed to investigate possible anti-tümör and apoptotic effects of exogen klotho in human colorectal adenocarcinoma cells (HT-29) and healthy colon cells (CCD 841 CoN). The WST-8 test was used to determine the half-maximum inhibitory concentration (IC50) of the klotho protein. AO-PI fluorescent staining techniques and Annexin V-PI flow cytometry was utilized to observe and detect the apoptosis of cancer cells induced by klotho. Our results demonstrated that klotho had a cytotoxic effect against colorectal adenocarcinoma cells in a dose-dependent manner. Our Annexin V-PI flow cytometric and AO-PI fluorescent analyses showed that klotho induced quantitative and morphological changes that indicate apoptotic induction in the human colorectal adenocarcinoma. This study results proved for the first time that klotho may be an effective potential therapeutic agent that may be used in adjuvant therapy in human colorectal adenocarcinoma it does not affect selectively healthy colon cells and but leading cancer cells to apoptosis.

Enhancement of Apo2L/TRAIL signaling pathway receptors by the activation of Klotho gene with CRISPR/Cas9 in Caco-2 colon cancer cells.

Human Klotho gene has many known functions such as anti-aging and anti-tumor, and decreased expression of this gene causes malignant formations in most types of cancer, including colon cancer. Interacting with TRAIL death receptors (DR4 and DR5) induces an apoptotic effect in cancer treatments by reducing the proliferation of cancer cells. The present study aimed to investigate downstream effect of overexpression of Klotho gene, which is known to have an antitumor effect on resistant human colon cancer cells, by examining its action on TRAIL death and decoy (DcR1 and DcR2) receptors for the first time. For this purpose, upregulation of human Klotho gene was achieved with CRISPR/Cas9-mediated system in resistant human colon cancer Caco-2 cells. To determine the effect of upregulation of Klotho gene on cancer cells evaluations with flow cytometry, WST-8, qRT-PCR, ELISA, and immunohistochemical analysis were performed. Then, Klotho gene was knocked out and its apoptotic effect was tested to find out whether it is due to overexpression of Klotho gene or not. Our results indicate that overexpression of Klotho gene in Caco-2 cells via CRISPR/Cas9-sensitized TRAIL death receptor DR4 suppresses the proliferation of cells by leading to apoptosis. Thus, this study conducted on apoptosis-resistant colon cancer cells may bring new insights about the role of Klotho gene in colon cancer.

The role of prokineticins in recurrent implantation failure.

The aim of the present study was to investigate the expression patterns of prokineticins (PROK) and prokineticin receptors (PROKR) in the endometrium of women with recurrent implantation failure (RIF). Fifteen (15) women with RIF and 15 fertile controls were enrolled in this study. Endometrial samples were taken from study participants with an endometrial biopsy cannula during the implantation window. Real time polymerase chain reaction and immunohistochemistry were used to determine PROK/PROKR mRNA expression and protein localization, respectively. PROK1 mRNA levels were 6.09 times higher compared to endometrial samples obtained from women with RIF than in samples obtained from fertile controls, whereas PROKR1 mRNA levels were 2.46 times lower in endometrial samples obtained from women with RIF than in samples from fertile controls. In addition, decreased PROKR1 was supported by immunohistochemistry analysis at protein level. There was no statistically significant difference between women with RIF and fertile controls regarding PROK2 and PROKR2 levels. Altered expression of the PROK1/PROKR1 system could be one of the numerous abnormalities in the endometrium of women with RIF.

The investigation of immunomodulatory effects of adipose tissue mesenchymal stem cell educated macrophages on the CD4 T cells.

Mesenchymal stem cells (MSCs) are strong immunomodulatory cells investigated in numerous clinical studies on fatal pathologies, such as graft versus host disease and autoimmune diseases; e.g., systemic lupus erythematosus, Crohn's disease, and ulcerative colitis. Macrophages are one of the critical cells linking the innate and adaptive immune system, and it has been shown that MSCs can differentiate between pro-inflammatory M1 phenotype and anti-inflammatory M2 phenotype of macrophages. However, it has not yet been fully clarified whether these differentiated macrophages are functional. In this study, we compared the immunomodulatory effects on the CD4 T cells of M1, M2a and M2c macrophages with the macrophages that directly and indirectly cultured with MSCs. We analyzed the changes in CD14, CD64, CD80, CD163 and CD200R expression to evaluate macrophage phenotypes, and the changes in CD4, IFN-g, IL-4, IL-17a and FoxP3 expression to evaluate T helper subsets using the FACS method. The changes in IL-1b, IL-4, IL-10, IL-12p70, IL-17a and IFN-g in the media supernatants were analyzed using the Luminex method. We also performed WST-1 and Caspase-3 ELISA analyses to observe the proliferation and apoptosis status of the T cells. MSCs were found to differentiate macrophages into a distinctive phenotype, which was close to the M2c phenotype, but was not considered as an M2c cell due to the low expression of CD163, a characteristic marker for M2c. While MEM-D, MEM-ID and MSCs showed similar inhibitory effects on the Th2 and Th17 cells, the most significant increase in Treg cell frequencies was seen in MEM-D cells. Macrophages can alter their phenotypes and functions according to the stimuli from the environment. The fact that macrophages educated with MSCs suppressed the production of all the cytokines we evaluated even after the removal of MSCs suggests that these cells may be differentiated by MSCs into a suppressive macrophage subgroup. However, the Treg cell activation caused by direct interactions between MSCs and macrophage cells may be the most prominent observation of this study compared to previous work. As a result, according to our data, the interactions between MSCs and macrophages may lead to differentiation of macrophage cells into an immunosuppressive phenotype, and these macrophages may suppress the T lymphocyte subgroups at least as effectively as MSCs. However, our data obtained from in vitro experiments should be supported by future in vivo studies.

Erythropoietin Protects the Kidney by Regulating the Effect of TNF-α in L-NAME-Induced Hypertensive Rats.

BACKGROUND/AIMS: Hypertension is the leading cause of death worldwide. Chronic high blood pressure induces inflammation. Tumor necrosis factor (TNF)-α plays a major role in inflammation and also depresses the synthesis of erythropoietin, which exerts protective effects on tissue; however, the mechanism is still unclear. We investigated the protective effect of erythropoietin against tissue damage caused by hypertension in the kidney and whether this effect was suppressed by TNF-α. METHODS: First, we detected the optimum chronic dose for darbepoetin-α (Depo), which is a long-acting erythropoietin analog for rats. We separated 60 female adult rats into 6 groups: control, Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME), L-NAME+Depo, L-NAME+Remicade (an anti-TNF-α antibody), L-NAME+Depo+Remicade, Depo, and control. After 1 month of treatment, we measured cardiovascular parameters, took blood samples, sacrificed the rats, and removed kidneys for analyses. RESULTS: The apoptotic index and the plasma and kidney mRNA levels of TNF-α increased in the L-NAME group and decreased in all other treatment groups. Macrophage accumulation increased in the L-NAME and L-NAME+Remicade groups, while it decreased in the Depo group. The mRNA abundance of TNF receptor 1 (TNFR1) decreased slightly in the Depo group and TNFR2 increased significantly in the same group. CONCLUSION: Erythropoietin protects kidney tissue against hypertension by preventing the apoptotic effects of TNF-α by blocking macrophage accumulation, decreasing TNF-α levels, and switching the TNF-α receptors from the apoptotic receptor TNFR1 to the proliferative receptor TNFR2.

Prenatal alcohol-induced neuroapoptosis in rat brain cerebral cortex: protective effect of folic acid and betaine.

PURPOSE: Alcohol consumption in pregnancy may cause fetal alcohol syndrome (FAS) in the infant. This study aims to investigate prenatal alcohol exposure related neuroapoptosis on the cerebral cortex tissues of newborn rats and possible neuroprotective effects of betaine, folic acid, and combined therapy. METHODS: Pregnant rats were divided into five experimental groups: control, ethanol, ethanol + betaine, ethanol + folic acid, and ethanol + betaine + folic acid combined therapy groups. We measured cytochrome c release, caspase-3, calpain and cathepsin B and L. enzyme activities. In order to observe apoptotic cells in the early stages, TUNEL method was chosen together with histologic methods such as assessing the diameters of the apoptotic cells, their distribution in unit volume and volume proportion of cortical intact neuron nuclei. RESULTS: Calpain, caspase-3 activities, and cytochrome c levels were significantly increased in alcohol group while cathepsin B and L. activities were also found to be elevated albeit not statistically significant. These increases were significantly reversed by folic acid and betaine + folic acid treatments. While ethanol increased the number of apoptotic cells, this increase was prevented in ethanol + betaine and ethanol + betaine + folic acid groups. Morphometric examination showed that the mean diameter of apoptotic cells was increased with ethanol administration while this increase was reduced by betaine and betaine + folic acid treatments. CONCLUSION: We observed that ethanol is capable of triggering apoptotic cell death in the newborn rat brains. Furthermore, folic acid, betaine, and combined therapy of these supplements may reduce neuroapoptosis related to prenatal alcohol consumption, and might be effective on preventing fetal alcohol syndrome in infants.

Efficiacy of resveratrol and quercetin after experimental spinal cord injury.

BACKGROUND: The aim of this study was to investigate the effect of natural antioxidants resveratrol and quercetin on oxidative stress and secondary cell damage in rats with acute spinal cord injury. METHODS: In this experimental study, 42 male Sprague-Dawley rats were used. Spinal cord injury was performed with clip compression method at level of T4-5. The study was conducted using 6 groups: control, trauma, trauma and solvent, trauma and resveratrol, trauma and quercetin, and trauma with combined resveratrol and quercetin. All rats were euthanized 48 hours after the procedure. Effects of resveratrol and quercetin on serum and tissue total antioxidant capacity and paraoxanase activity level were examined. RESULTS: Compared to trauma group, there was a significant increase in total antioxidant capacity and paraoxanase activity level in resveratrol, quercetin, and combined treatment groups. There was no significant difference between resveratrol and quercetin groups with regard to total antioxidant capacity and paraoxanase activity level. Total antioxidant capacity and paraoxanase activity level were significantly higher in solvent group than trauma group. In histopathological evaluation, there was a decrease in polymorphonuclear leukocyte infiltration in solvent, resveratrol, quercetin, and combined treatment groups. CONCLUSION: Biochemical and histological staining results of present study showed that resveratrol and quercetin may be effective in preventing secondary damage in spinal cord injury.

The relationship between serum thyroid hormone levels, subclinical hypothyroidism, and coronary collateral circulation in patients with stable coronary artery disease.

OBJECTIVE: Thyroid disease is a common endocrine disease with important effects on the cardiovascular system. As an adaptive response to myocardial ischemia, coronary collateral circulation (CCC) plays an important role in obstructive coronary artery disease (CAD). The association between serum thyroid hormone levels and development of CCC was investigated in the present study. METHODS: In total, 430 consecutive patients who underwent coronary angiography procedure and had documented total occlusion in at least 1 major coronary artery were investigated retrospectively. Degree of CCC was classified according to Cohen-Rentrop method. Serum free triiodothyronine (FT3), free thyroxine (FT4) and thyroid-stimulating hormone (TSH) were assessed by the chemiluminescence immunoassay technique. RESULTS: In spite of diabetes mellitus (p=0.019), smoking (p<0.001), and TSH (p<0.001), FT3 (p<0.001), FT4 (p=0.015), and subclinical hypothyroidism (SCH) (p<0.001) ratios were significantly different between groups. In regression analysis, SCH (p=0.024), DM (p=0.021), smoking (p<0.001), and heart failure (p=0.029) were independent predictors of poor CCC development in multivariate model 1. When regression analyses were performed based on multivariate model 2, TSH (p<0.001), FT3 (p<0.001), heart failure (p=0.022), smoking (p<0.001), and hyperlipidemia (HPL) (p=0.046) were independent predictors of poor CCC development. CONCLUSION: In addition to traditional risk factors, SCH, higher serum TSH, and lower FT3 levels were associated with development of poor CCC in patients with obstructive CA.

[The effect of ligustrazin in intestinal ischemia reperfusion injury generated on rats]. / Siçanlarda olusturulan ince bagirsak iskemi reperfüzyon hasarinda ligustrazin'in etkisi.

BACKGROUND: Intestinal ischemia is a serious and common clinical status. It develops as result of superior mesenteric artery (SMA) obstruction caused by many etiologic factors. Sepsis and multiple organ failure could develop following intestinal ischemia. The present study aimed to investigate the effects of ligustrazin, which has a vasodilator impact on intestinal ischemia. METHODS: Forty male Wistar rats were divided into three groups randomly. Sham operation was performed on Group S (n=7); mesenteric ischemia and then 60 minutes reperfusion of the intestine process was performed on Group MI (n=7); mesenteric ischemia and then 60 minutes reperfusion of the intestine process was performed and 80 mg/kg ligustrazin was administrated intraperitoneally on Group MI+L (n=7). Intestinal tissue samples were taken for tissue MDA, SDO and nitric oxide (NO) levels, and ileum and jejunum samples were taken for histopathologic examination. RESULTS: Tissue MDA levels and tissue NO levels of Group MI-L was determined to have significantly decreased. Tissue SOD levels were found similar to Group S. Chiu classification score of the jejunum and ileum was determined to have decreased in Group MI-L compared to Group MI. DISCUSSION: As a result of this study, Ligustrazin was found to adjust lipid peroxidation in biochemical parameters during mesenteric I-R and decrease the severity of damage of I-R on the histopathological scores of the jejunum and ileum.

The effect of iloprost and N-acetylcysteine on skeletal muscle injury in an acute aortic ischemia-reperfusion model: an experimental study.

OBJECTIVE: The objective of this study was to examine the effects of iloprost and N-acetylcysteine (NAC) on ischemia-reperfusion (IR) injuries to the gastrocnemius muscle, following the occlusion-reperfusion period in the abdominal aorta of rats. MATERIALS AND METHODS: Forty male Sprague-Dawley rats were randomly divided into four equal groups. Group 1: control group. Group 2 (IR): aorta was occluded. The clamp was removed after 1 hour of ischemia. Blood samples and muscle tissue specimens were collected following a 2-hour reperfusion period. Group 3 (IR + iloprost): during a 1-hour ischemia period, iloprost infusion was initiated from the jugular catheter. During a 2-hour reperfusion period, the iloprost infusion continued. Group 4 (IR + NAC): similar to the iloprost group. FINDINGS: The mean total oxidant status, CK, and LDH levels were highest in Group 2 and lowest in Group 1. The levels of these parameters in Group 3 and Group 4 were lower compared to Group 2 and higher compared to Group 1 (P < 0.05). The histopathological examination showed that Group 3 and Group 4, compared to Group 2, had preserved appearance with respect to hemorrhage, necrosis, loss of nuclei, infiltration, and similar parameters. CONCLUSION: Iloprost and NAC are effective against ischemia-reperfusion injury and decrease ischemia-related tissue injury.

Changes in carotid intima-media thickness over two years in patients on haemodialysis.

OBJECTIVE: To evaluate the progression of carotid intima-media thickness (CIMT) and to search for possible associations between these changes and other risk factors of atherosclerosis for 2 years in stable patients with chronic renal failure (CRF) on haemodialysis (HD). METHODS: Study population consisted of 22 patients with newly diagnosed CRF. All patients underwent B-mode ultrasonography of common carotid artery for estimating CIMT and the presence of plaques before and after the first HD session (mean 24.22 +/- 2.14 months). The differences in CIMT before and after long-term HD treatment were compared. Acute phase proteins, calcium-phosphate balance and lipid profile were assessed and anthropometric parameters were measured. RESULTS: Mean age was 55 +/- 13 years and 10 (45%) of the patients were female. After long-term HD treatment, (mean 24.22 +/- 2.14 months) the mean value for CIMT (0.57 +/- 0.08 mm) was significantly lower than that at baseline (0.68 +/- 0.12 mm) (p = 0.02). Only male gender and smoking were correlated with baseline CIMT. After long-term HD treatment, age, total cholesterol, LDL cholesterol, and triglyceride were related with CIMT. Diabetes and smoking were correlated with CIMT. Presence of plaque before HD only correlated with creatinine level and after long-term HD treatment only correlated with total cholesterol level. CONCLUSION: We found that CIMT was significantly decreased 2 years after starting HD. An association between CIMT and other atherosclerotic risk factors (such as age, cholesterol, triglyceride etc.) could not be determined due to a small sample size.

[The frequency of aspirin resistance by a modified thrombelastography method and its relationship with clinical and laboratory parameters in patients with stable coronary artery disease]. / Kararli koroner arter hastaligi olanlarda modifiye tromboelastogram yöntemi ile aspirin direnci sikligi ve klinik ve laboratuvar bulgularla iliskisi.

OBJECTIVES: Aspirin is the cornerstone of antiplatelet therapy in cardiovascular medicine. However, aspirin resistance has been demonstrated in 0.4% to 83.3% of aspirin-receiving patients. The aim of this study was to investigate the frequency of aspirin resistance using a modified thrombelastography (mTEG) method and related clinical and biochemical parameters in patients with stable coronary artery disease (CAD), who received 100 mg/day aspirin. STUDY DESIGN: The study included 168 patients (115 males, 53 females; mean age 60±8 years) with stable CAD, receiving aspirin at a dose of 100 mg/day. Aspirin responsiveness was determined using mTEG, where aspirin resistance was defined as arachidonic acid-induced whole blood platelet aggregation inhibition (PAI) of less than 50%. RESULTS: Aspirin resistance was detected in 27 patients (16.1%). Platelet aggregation inhibition showed negative correlations with hyperlipidemia, smoking, spironolactone use, systolic blood pressure, pulse pressure, and total cholesterol and fibrinogen levels. In multivariate regression analysis, only fibrinogen level (OR=1.063, p=0.010) and pulse pressure (OR=1.197, p=0.023) were found to be independent indicators of aspirin resistance and PAI. In ROC analysis, cut-off values of 50 mmHg for pulse pressure and 400 mg/dl for fibrinogen level predicted aspirin resistance with 88.9% and 74% sensitivity and 64.4% and 68% specificity, respectively. CONCLUSION: Our findings suggest that measurements of fibrinogen level and pulse pressure may be used as easy and reliable methods in predicting aspirin resistance.

Anti-apoptotic and antioxidant effect of leptin on CCl4-induced acute liver injury in rats.

The aim of this study is to investigate the effect of leptin in rats on carbon tetrachloride (CCl(4)) induced acute liver damage using immunohistochemical methods for apoptosis and biochemical parameters. In this experimental study, 18 Spraque-Dawley rats were divided into three groups viz; control, CCl(4) and CCl(4)+leptin treatment. 0.8 ml/kg olive oil was administered intraperitoneally (i.p.) to the control group and 0.8 ml/kg CCl(4) (1:1 dissolved in olive oil) was administered i.p. to the CCl(4) and CCl(4)+leptin treatment groups, respectively. After 6 h of administrating CCl(4), CCl(4)+leptin treatment group was given i.p. leptin (10 µg/kg). Twenty-four hours after administrating CCl(4) all of the groups were euthanized. Biochemical assessments were performed using serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), plasma tumor necrosis factor alpha (TNF-α) levels and tissue malondialdehyde (MDA), and TNF-α levels. Histological assessments were then performed using Hematoxylin&Eosin (H&E) staining in light microscope and apoptosis assessment using Terminal Transferase dUTP Nick End Labeling (TUNEL)-staining. Serum AST, ALT, ALP and plasma TNF-α levels, tissue MDA and TNF-α levels had all increased in CCl(4) group, but were found to be significantly decreased in CCl(4)+leptin treatment group. Moreover, TUNEL-positive cell counts in liver had significantly increased in CCl(4) group, but decreased in CCl(4)+leptin treatment group (P < 0.05). The results of our study the biochemical, histological and TUNEL-staining showed that leptin has treatment effects on liver CCl(4) induced injury. It plays a role as a potent free radical scavenger, a powerful antioxidant and it also has anti-apoptotic effects.

Protective effect of vasoactive intestinal peptide on testicular torsion-detorsion injury: association with heparin-containing mast cells.

OBJECTIVES: To elucidate the action of vasoactive intestinal peptide (VIP) on detorsion injury and the heterogeneity of mast cells in the testes of rats. METHODS: Prepubertal male Sprague-Dawley rats were used in six groups. Group 1 was the control group (sham operation); group 2 had 2 hours of torsion; group 3, 2 hours of torsion and 1 hour of detorsion after administration of saline; group 4 had 2 hours of torsion and 4 hours of detorsion after administration of saline; group 5, 2 hours of torsion and 1 hour of detorsion after administration of intraperitoneal VIP (25 ng/kg); and group 6, 2 hours of torsion and 4 hours of detorsion after intraperitoneal VIP. The 2 hours of torsion was created by rotating the right testis 720 degrees in a clockwise direction. VIP (25 ng/kg) was injected intraperitoneally 1 minute before the 1 and 4 hours of detorsion. At the end of the experiment, catalase enzyme activity was measured polarographically, and superoxide dismutase, malondialdehyde, and protein were measured spectrophotometrically. Nitric oxide was measured by capillary electrophoresis in the testicular tissue. Routine histologic examination of testicular mast cells was done under light microscopy; the histochemistry was also analyzed. RESULTS: Torsion significantly induced oxidative stress, mast cell degranulation, and tissue damage. Detorsion attenuated oxidative stress without any diminution of the histologic damage to the tissue. VIP significantly protected the testicular tissue from detorsion injury. It also inhibited mast cell activity while increasing the heparin content. CONCLUSIONS: VIP can protect testicular tissue from detorsion injury. Heparin-containing mast cells seem to be important mediator cells for this protection.