Colorectal Cancer: An Update on Treatment Options and Future Perspectives.

Throughout the years, colorectal cancer has steadily become a global health problem. While other types of cancers have seen a decline in cases because of screening and vaccination programs, colorectal cancer has risen become the third most diagnosed cancer worldwide and, more worryingly, the second leading cancer-related cause of death. The introduction of targeted therapy has been widely considered a major paradigm shift in the treatment of colorectal cancer, which agents such as bevacizumab and cetuximab quickly becoming mainstay options in the treatment of locally advanced or metastatic disease. However, this type of treatment has also shown its limitations, with limited or no benefit for a large portion of the patients. With more and more knowledge being gathered on the molecular mechanisms which govern the malignant phenotype presented by colorectal cancer, scientists are engaged in a continuous effort to develop new therapies based on these discoveries.

A Statistical Analysis of Risk Groups in Colorectal Cancer Patients.

Colorectal cancer (CRC) is considered a major global health concern due to an increasing number of new cases and cancer-related deaths each year, strong link to dietary habits prevalent in middle and high-income countries and limited therapeutic options especially in locally-advanced and metastatic settings. To counter this growing problem, the scientific community has strived to underpin the major molecular mechanisms behind the aggressive phenotype displayed by CRC and also develop new agents to selectively target and inhibit these core drivers. This evolution has allowed the separation of patients according to different risk groups in concordance with epidemiological parameters alongside novel biomarkers such as gene alterations, protein overexpression and aberrant signaling pathways. In this study we included 20 patients who underwent colonoscopy and were later received histopathologic confirmation of CRC. The statistical anamnestic data obtained from the patients (age, gender, home distribution, signs and symptoms) was corroborated with the results obtained from the histopathologic and immunohistochemical analysis of the samples obtained via colonoscopy. The average age was 63.8 years, the male: female ratio was 2.33 and the origin of 2/3 of the patients was urban and the most encountered symptoms were transit disorders (75%). In terms of colonoscopy results, the majority of tumors were found on the rectum (85%), 90% of tumors were adenocarcinomas, having a vegetant aspect in 60% of the cases and a moderate degree of differentiation in 50% of situations.

Molecular phenotypes of the human kidney: Myoid stromal cells/telocytes and myoepithelial cells.

The connective stromal and epithelial compartments of the kidney have regenerative potential and phenotypic flexibility. A few studies have shown that cells appertaining to both compartments can exhibit myoid phenotypes. The purpose of our study was to investigate the myoid pattern of kidney and its association with the kidney niches containing stromal cells/telocytes (SC/TCs). We performed an immunohistochemical study using a panel of endothelial, myoid, mesenchymal and stem/progenitor markers, namely CD31, CD34, CD105 (endoglin), CD117/c-kit, nestin, desmin, α-smooth muscle actin (α-SMA) and the heavy chain of smooth muscle myosin (SMM). We used histologically normal kidney samples, obtained after nephrectomy, from nine adult patients. The capsular SC/TCs had a strong CD34 and partial nestin and CD105 immunopositivity. Subcapsular and interstitial SC/TCs expressed c-kit, nestin, CD105, but also α-SMA and SMM, therefore having a myoid phenotype. The endothelial SC/TCs phenotype was CD31+/CD34+/CD105+/nestin±/SMM±/α-SMA±. All three myoid markers were expressed in periendothelial SC/TCs. We also found a scarce expression of nestin in parietal epithelial cells of Bowman's capsule, and in podocytes. In epithelial cells, we found a positive expression for CD31, CD117/c-kit, desmin, CD34, SMM, and CD105. In epithelial tubular cells, we found a predominant basal expression of the myoid markers (SMM and desmin). In conclusion, myoepithelial tubular cells, myoid endothelial cells and myoid SC/TCs are normal constituents of the kidney.

Urinary Bladder Tumors Clinical and Statistical Retrospective Study.

Nowadays, urinary bladder cancer represents a major health problem, due to very high medical and social costs. This disease affects mainly the elderly. We performed a study on 1073 patients admitted to the Urology Clinic within the Emergency Clinical Hospital of Craiova, between 2013-2015 with bladder cancer. Of the 1073 cases, 741(69.06%) were diagnosed in men, and 332 (30.94%) were found in women, the men/ women ratio being of 2.23/1. The highest incidence of bladder tumors was recorded in individuals aged between 60 and 79 years old. In this age group, there were admitted 734 patients with bladder cancer, representing 68.44%. Regarding the tumor recurrence, out of 1073 bladder tumors, a number of 608 (56.66%) patients were diagnosed with primary tumors, while a number of 465 (43.34%) patients presented recurrent tumors. Of the symptoms presented by the patients, the most frequent were haematuria (present in about 87% of the patients), pollakiuria (present in 64% of the patients), dysuria (present in 55% of the patients) and urinary infections (present in about 23% of the patients).

Repeated Events of Acute Ischemic Stroke in a Patient with Essential Thrombocythemia.

Essential thrombocythemia (ET) represents a risk factor for ischemic stroke, although it is a rare cause. Chronic myeloproliferative disorder is associated with proliferation of megakaryocytes sustained increases circulating platelet count. Essential thrombocythemia cause is not known, yet, many patients suffering from this disease may have no symptoms for a long time. Early detection is necessary because it may recur frequently thrombosis if not treated properly. We present a case of a 72 year old man with a history of three stroke events. The clinical diagnostic procedure revealed an increased platelet count was 961000/ml, and these cerebrovascular events were the first manifestation of essential thrombocythemia.

Correlation between Immunohistochemical Subtype and Clinicopathological Features in Patients with Diffuse Large B-cell Lymphoma.

The aim of this study was to establish correlations between certain clinical, biological, therapeutic factors and diffuse large B-cell lymphoma (DLBCL) subtypes. For this purpose, between January 2007 and December 2016 a total number of 97 patients with de novo diffuse large B-cell lymphoma were analyzed. Patients with a high prognostic index and non-GCB DLBCL positively correlated and exhibited lower survival rates than low IPI, GCB patients. IPI scoring system and cell-of-origin classification should be used together as a single valid prognostic evaluation tool for DLBCL.

Therapeutic Options in Diffuse Large B-Cell Lymphoma - A Retrospective Study and Review of the Literature.

The aim of this retrospective study was to assess the differences between standard R-CHOP and other Rituximab-associated chemotherapy (R-miniCHOP and R-CHOEP) regimens in terms of survival and potential adverse effects. The six-month survival outcomes of 94 diffuse large B-cell lymphomas (DLBCL) patients indicated no statistical difference between overall survival and disease-free survival in the two subgroups. The biological response to therapy (blood count, LDH levels) was similar in both subgroups. Despite having different clinical indications, R-miniCHOP and R-CHOEP provide viable therapeutic alternatives to the standard R-CHOP regimen.

Embryonic hematopoietic stem cells and interstitial Cajal cells in the hindgut of late stage human embryos: evidence and hypotheses.

There have been few studies on human embryos describing a specific pattern of hindgut colonization by hematopoietic stem cells (HSCs) and interstitial Cajal cells (ICCs). We aimed to study CD34, CD45 and CD117/c-kit expression in late stage human embryos, to attain observational data that could be related to studies on the aorta-gonad-mesonephros (AGM)-derived HSCs, and data on hindgut ICCs. Antibodies were also applied to identify alpha-smooth muscle actin and neurofilaments. Six human embryos of 48-56 days were used. In the 48 day embryo, the hindgut was sporadically populated by c-kit+ ICCs, but, in all other embryos, a layer of myenteric ICCs had been established. Intraneural c-kit+ cells were found in pelvic nerves and vagal trunks, suggesting that the theory of Ramon y Cajal assuming that ICCs may be primitive neurons may not be so invalid. Also in the 48 day embryo, c-kit+/CD45+ perivascular cells were found along the pelvic neurovascular axes, suggesting that not only liver, but also other organs could be seeded with HSCs from the AGM region. CD45+ cells with dendritic morphologies were found in all hindgut layers, including the epithelium. This last evidence is suggestive of an AGM contribution to the tissue resident macrophages and could be related to processes of sprouting angiogenesis which, in turn, have been found to be guided by filopodia of endothelial tip cells. Further studies on human embryonic and fetal material should be performed to attempt to clarify whether the hindgut colonization with HSCs is a transitory or definitive process.

A Case of Bilateral Aseptic Necrosis of the Femoral Head.

Aseptic necrosis of the femoral head is a disease whose etiology is not completely elucidated and generally affects young adults aged between 30 and 50 years. In a significant number of patients bilateral disease occurs, which makes detection in its early stages constitute an important objective. We present the case of a male patient, aged 23 years, with the following risk factors: smoking and chronic alcohol consumption, who is diagnosed with aseptic necrosis of the left femoral head, ARCO stage IV, and in just six months after the diagnosis and hip arthroplasty, he suffers an injury which leads to the same diagnosis in the contralateral hip. We want to emphasize that for all patients with a high index of suspicion there should be an MRI examination, because the plane radiographs or CT are most often not relevant in detecting early signs of this condition. Diagnosis of aseptic necrosis of the femoral head in the early stages is a necessity in order to obtain an optimal result of conservative treatment.

Squamous cell carcinoma of the oral cavity: clinical and pathological aspects.

Oral cavity cancer is a public health problem as the sixth leading cause of cancer worldwide. Most tumor lesions are detected in stage III and IV, leading to a poor prognosis, five-year survival rate ranging between 10% and 40%. Oral cancer etiology is multifactorial, known still incomplete. The main etiopathogenic factors are exposure to cigarette smoke and alcohol consumption. We conducted a retrospective study of oral cavity tumors hospitalized in 2008-2012 in Oral and Maxillofacial Surgery Clinic of the Emergency County Hospital of Craiova, Romania. Of 143 tumors of the oral cavity, 125 were malignant, and of these, 115 (92%) were represented by squamous cell carcinoma. Tumor lesions were more common in males (69%), patients from rural areas (64%) and those over 50-year-old (87.71%).

The effect of ultraviolet radiation on the cornea - experimental study.

Ultraviolet (UV) radiation in high doses may have harmful effects on the eye. The sources of UV radiation are the sun, as well as some artificial sources such as UV lamps or voltaic arcs. Chronic exposure to UV can cause damage to the anterior pole of the eye, ranging from minor (pterygium) to serious photokeratitis. In our study, we applied a UV dose of 6.5 J/cm(2) in the wavelength range of 290-400 nm, for five consecutive days per rat anterior pole of the eye. Seven days after the last dose of radiation, the animals were sacrificed, harvesting both the irradiated and the non-irradiated eye. Histological and immunohistochemical examination of the lesions revealed that the greatest damage to the epithelium was recorded prior to and 2/3 of the remaining corneal stroma. The epithelial lesions we found varied from pseudokeratosis and detachment of the Bowman epithelium membrane to deep epithelial necrosis. Within the corneal stroma, we observed the formation of interstitial edema with disruption of the collagen structure. We also noticed the presence of an inflammatory infiltrate composed mainly of lymphocytes and CD68+ and CD163+ macrophages, as well as the occurrence of vascular devices. These consisted of angiogenesis capillaries with structured wall composed mainly of endothelial CD34+ precursor cells and a basal membrane rich in collagen IV fibers.

Bony metaplasia in a caecum adenocarcinoma.

Ectopic bone formation in the digestive tract is a very rare phenomenon in pathology. Most cases of bone metaplasia were found in distal colon tumors, rectum and sigmoid respectively. We present a case of well-differentiated adenocarcinoma of the caecum (the second case of literature) associated with bone metaplasia, which debuted atypical, with symptoms of infectious enterocolitis, in a 72-year-old female patient, with no history of digestive pathology. Highlighting bone metaplasia was achieved by current histological techniques. Patient's evolution was favorable with surgical treatment and chemotherapy.

Macroscopic and microscopic findings in avascular necrosis of the femoral head.

UNLABELLED: The avascular necrosis of the femoral head is an illness induced by the cutoff of blood flow to the femoral head and it affects mostly young adults between the ages of 30 and 50 years, raising therapeutic and diagnostic issues. Many risk factors are incriminated in the development of avascular necrosis of the femoral head like: trauma, chronic alcohol consumption, smoking, administration of corticosteroid drugs, most of the cases are considered to be idiopathic. The main goal of our paper is to describe the macroscopic and microscopic variations of the bone structure, which occur in patients with avascular necrosis of the femoral head. MATERIALS AND METHODS: The biological material needed for our study was obtained following hip arthroplasty surgery in 26 patients between the ages of 29 and 59 years, which previously were diagnosed with avascular necrosis of the femoral head and admitted in the Orthopedics Department of the Emergency County Hospital of Craiova (Romania) between 2010 and 2011. From a macroscopic point of view, we found well defined areas of necrosis, most of which were neatly demarcated of the adjacent viable tissue by hyperemic areas, loss of shape and contour of the femoral head and transformations of the articular cartilage above the area of necrosis. When examined under the microscope, we found vast areas of fibrosis, narrow bone trabeculae, obstructed blood vessels or blood vessels with clots inside, hypertrophic fat cells, bone sequestration but also small cells and pyknotic nuclei. The microscopic and macroscopic findings on the femoral head sections varied with the patients and the stage of the disease.

Histological and immunohistochemical changes of the myocardium in dilated cardiomyopathy.

Dilated cardiomyopathy is a major cause of heart failure and a major cause of morbidity and mortality. It is a multifactorial disease that includes both hereditary and acquired forms. It is estimated that around 20-35% of patients with dilated cardiomyopathy have hereditary forms. It is the third most common cause of heart failure and the most common cause of heart transplant. Dilated cardiomyopathy can be a secondary condition of many diseases such as coronary heart disease, diabetes, pheochromocytoma, infections, malnutrition, ingestion of toxic substances (alcohol, cocaine), ingestion of chemotherapeutic drugs, autoimmune diseases. In our study, we aimed to describe the changes of myocardial cells and interstitial connective tissue in patients clinically diagnosed with alcoholic dilated cardiomyopathy. The material studied consisted of heart fragments sampled from the left ventricle (LV) during necropsy from a total of 28 patients, aged between 58 and 73 years, with a clinical and laboratory diagnosis of dilated cardiomyopathy, hospitalized in the Cardiology Center of the Emergency County Hospital of Craiova in 2009 and 2010. In dilated cardiomyopathy, myocardial muscle fibers appeared slightly elongated or wavy, with hypochromatic, heterogeneous, vacuolar sarcoplasm, by a decrease of myofibril numbers. Lipofuscin granules were frequently seen in the sarcoplasm. Nuclear changes were consistent with sarcoplasmic alterations. Changes of the interstitial connective tissue were sometimes extensive and sometimes barely noticeable. The most common alteration of this structure was the onset and development of a mainly perivascular collagen fibrillogenetic process.

Chronic gastritis with intestinal metaplasia: clinico-statistical, histological and immunohistochemical study.

Chronic gastritis has a high incidence in adults, causing progressive destruction of glandular structures, favoring the development of gastric atrophy. The association of chronic gastritis with intestinal type metaplasia of gastric mucosa has a poor outcome as intestinal metaplasia is regarded as a precancerous lesion. Metaplasia is common in patients with Helicobacter pylori infection and also heavy smokers. The aim of our study was to evaluate the relationship between chronic gastritis and intestinal metaplasia. The study was conducted on a total of 1218 patients, aged between 5 and 90 years, who presented for dyspeptic disorders in the period 2007-2010 and were examined clinically and endoscopically. During the gastroscopic examination, fragments of gastric mucosa were collected for the histopathological study and for highlighting the H. pylori infection. For the histopathological study, the Hematoxylin-Eosin and PAS-Alcian Blue stains were performed, while for the immunohistochemical study the anti-TAG72 and anti-PCNA antibodies were used. A diagnosis of gastritis was established in 615 patients, representing approximately 50.5% of all cases. Most cases with gastritis were found in people of middle age. Gastritis was present in almost all age groups, from teenagers to the elders. Of the 615 cases of gastritis, urease test was positive in 353 patients, representing approximately 57.40% of all patients with gastritis. Histopathological examination identified the presence of intestinal metaplasia in 61.60% of patients with chronic gastritis, mostly complete metaplasia. PCNA immunohistochemistry revealed that cell proliferation processes are intensified in intestinal metaplasia. This study highlights the importance of chronic gastritis, intestinal metaplasia, and H. pylori infection in the etiopathogeny of gastric cancer.

Macrophage response in experimental third-degree skin burns treated with allograft. Histological and immunohistochemical study.

Macrophages are some of the innate immune cells with a central role in inflammatory and immune responses. Studies in the last 20 years have shown that these cells have a particular influence in the reparative processes also. Our aim in this study was to evaluate the macrophage response in third-degree skin burns treated with allograft in an experimental model. Macrophages were specifically highlighted by immunohistochemical staining with anti-CD68 antibody. In the first evolutive part of the reparatory process, macrophages rapidly increased both numerically and as a relative area with about 300%, and then decreased progressively along with the granulation tissue maturation. Macrophage overall response curve was similar in animals treated with allograft and in the control group (untreated), which leads us to believe that the allograft does not induce a more ample immune response that could be regarded as pathological.

Microscopic aspects of macrophage system cells in hemorrhagic stroke in humans.

Stroke is an important public health issue because it has high morbidity and mortality rates. In addition, it has one of the highest rates of disability in adults. Recent data show that macrophage system cells, especially microglial cells, are involved both in neuroprotective processes and in the neurotoxicity, depending on the type and extent of the brain damage. In our study, using histology and immunohistochemistry techniques, we evaluated the macrophage-type cell reaction in cerebral hemorrhage. We found that the number of CD68-positive cells increased 7-8 folds per square millimeter of cortical surface in the cerebral parenchyma adjacent to the hemorrhage. We identified a large number of perivascular-activated macrophages, in areas distant to the hemorrhage, showing that individuals with hemorrhagic stroke have profound and extensive alterations of the blood-brain barrier.

Fractal analysis differentiation of nuclear and vascular patterns in hepatocellular carcinomas and hepatic metastasis.

Hepatocellular carcinoma (HCC) currently represents the fifth most common cancer worldwide, while being the third leading cause of cancer death. Fractal analysis is a novel tool used in quantitative and qualitative image assessment. Vascular patterns and cellular nuclei particularities in tumoral pathology make ideal candidates for this technique. Our aim was to apply fractal analysis in quantifying nuclear chromatin patterns and vascular axels in order to identify differences between images of primary HCC, liver metastasis (LM) and surrounding normal liver tissue. Formalin-fixed, paraffin-embedded tissue sections from 40 cases of HCC and 40 LM of various origins were used. We performed Hematoxylin staining for nuclear chromatin as well as immunohistochemical staining for vascular patterns. High-resolution images were captured; nuclear and vascular morphologies were assessed on binarized skeleton masks using the fractal box counting method. Analysis was performed using the free, public domain Java-based image processing tool, ImageJ, which provided the fractal dimensions (FDs) for each studied element. Statistical analysis was performed using the ANOVA test with Bonferroni post-tests and t-tests for paired samples. Fractal analysis of vascular patterns clearly differentiated between tumoral tissue and normal surrounding tissue (p<0.01). Further analysis of nuclear FDs improved the specificity of these results, providing clear differentiation between pathological and normal tissue (p<0.01). When comparing primary HCC images with metastatic formations, we encountered statistically significant differences in nuclear chromatin assessment. However, blood vessels had a higher FD in primary tumors when compared with liver metastasis (p<0.05) and also allowed for a differentiation between primary liver tumors with and without neurodifferentiation. Fractal analysis represents a potent tool for discriminating between tumoral and non-tumoral tissue images. It provides accurate, quantifiable data, which can be easily correlated with the pathology at hand. Primary and metastatic liver tissue can be differentiated to some extent, however further studies, possibly including other variables (cellular matrix for instance) are needed in order to validate the method.

Pterygium: histological and immunohistochemical aspects.

INTRODUCTION: Pterygium represents a triangular conjunctivo-epithelial overgrowth, proliferating from the bulbar conjunctiva and covering the cornea, causing severe vision loss. It is an abnormal growth and differentiation of the conjunctive epithelial structures of the corneal limbus. Chronic exposures to solar ultraviolet radiation, heat and dust, as well as viral agents, are the most common pathogenic entities involved in its evolution. Recent studies linked pterygium with neoplastic proliferation, as ocular limbic stem cells and p53-protein expression are altered. MATERIALS AND METHODS: Our study was conducted on 84 fragments of pterygium, collected after surgery from patients admitted between 2008 and 2009 in the Departments of Ophthalmology of the Emergency County Hospital of Craiova and Hospital of Rovinari. Histological studies were performed by staining with Hematoxylin-Eosin, light green trichromic (Goldner-Szekely technique) and PAS-Hematoxylin. Immunohistochemistry highlighted the T-lymphocytes by using the CD3 antibody, B-lymphocytes by using the CD20 antibody and cells of the macrophage system using the CD68 antibody. The slides were analyzed under a 55I Nikon microscope, resulting pictures being captured with a 5 MP digital camera and digitally retrieved and enhanced using the dedicated NIS-Elements software. RESULTS: Histology showed the presence of a conjunctivo-epithelial structure, significantly different from the structure from which it developed. In 20% of the cases, the covering epithelium was similar to that of the bulbar conjunctival mucosa. In some cases, when pathogens were more active in the environment, the appearance of the covering epithelium of the membrane appeared pleomorphic, with dysplastic aspects, suggesting significant alteration of cell proliferation and differentiation. In approximately 75% of patients, we identified goblet cells in the surface epithelium. They appeared either isolated or associated in variable numbers, structures resembling intraepithelial glands. These particular cells synthesize and accumulate PAS-positive mucines rich in glycosaminoglycans, and are usually found in conjunctival epithelium. We observed a number of invaginations in the connective tissue underlying the epithelium, mostly formed by goblet cells, this giving the aspect of mucous glands, similar to the "glands of Henle". A highly developed vascular neoformation network, consisting of arterioles, venules and a very large number of capillaries can also be found in the connective tissue. Immunohistochemistry suggested that B-lymphocytes marginally take part in the immune response in pterygium. T-lymphocytes formed the majority of the mass of immune system cell present in connective tissue of the pterygium. Macrophage-type cells were distributed unevenly in the pterygium tissue, as the intensity of the inflammatory process varies depending on antigen levels. CONCLUSIONS: Pterygium shows significant changes both in the epithelium and in the underlying connective tissue. Immune cell infiltrate was diffuse, more abundant in areas with erosion of the covering epithelium.

Lymphatic and blood microvasculature organisation in pulmonary sarcoid granulomas.

Pulmonary sarcoid granulomas are characterised by their elective distribution along collecting lymphatics. However, relationships between granulomas and intralobular lymphatics or blood microvascularisation have not been investigated. Therefore, we undertook a specific analysis of blood capillaries and lymphatics supplying sarcoid granulomas to identify additional clues to understanding the pathophysiogenesis of these lesions. Six pulmonary samples were immunolabelled with D2-40, anti-CD34 and anti-CD31 antibodies, paying particular attention to the relationships between lymphatics and granulomas, and the pattern of blood microvessels supplying sarcoid lesions. A morphometric study of granulomas included their distance to lymphatics and a three-dimensional reconstruction of a granuloma in its lymphatic context. Intralobular granulomas were closely associated with lymphatics; apart from a few granulomas, blood capillaries stopped at the outer border of the fibrous ring surrounding granulomas, and perigranuloma capillaries were particularly scarce. Our observations of the lymphatic and blood microvascular environment of intralobular pulmonary sarcoid granulomas provide evidence for the critical role of lymphatics in the emergence of these lesions. Moreover, pulmonary sarcoid lesions could be considered avascular structures, thereby providing new insights into the understanding of the granuloma physiology and the distribution of blood-borne therapeutic agents.