Significance of epidermal growth factor receptor expression in breast cancer

Colorectal cancer is one of the most common cancers worldwide. The anticancer effect of Wolfberry (Lycium barbarum) polysaccharide (LBP) on colon cancer cells is largely unknown. To investigate the growth effect of LBP on human colon cancer cell and its possible mechanisms, human colon cancer SW480 and Caco-2 cells were treated with 100–1,000 mg/l LBP for 1–8 days. Cell growth was measured by MTT assay and crystal violet assay. Distribution of the cell cycle was analyzed by flow cytometry. Western blotting was used to indicate changes in the level of cyclins and cyclin-dependent kinases (CDKs). LBP treatment inhibited both colon cancer cell lines in a dose-dependent manner. At concentrations from 400 to 1,000 mg/l, LBP significantly inhibited the growth of SW480 cells (400 mg/l, P < 0.01; 800 and 1,000 mg/l, P < 0.001); while at concentrations from 200 to 1,000 mg/l, LBP significantly inhibited the growth of Caco-2 cells (200 mg/l, P < 0.05; 400–1,000 mg/l, P < 0.001). Crystal violet assay showed that LBP had a long-term anti-proliferative effect. More importantly, cells were arrested at the G0/G1 phase. The changes in cell-cycle-associated protein, cyclins, and CDKs were consistent with the changes in cell-cycle distribution. This is one of the first studies to focus on LBP-induced interruption of the cell cycle in human colon carcinoma cells. The results suggest that LBP is a candidate anticancer agent.     

Systemically available bone morphogenetic protein two and seven affect bone metabolism

Purpose

Bone morphogenetic protein (BMP)-2 and -7 are used in patients with long-bone fractures, nonunions and spinal fusions. It is unknown whether their potential systemic bioavailability following local bone administration might affect skeletal metabolism. To answer this question, we examined effects of systemically administered BMP-2 and -7 on bone in a newly developed rat model with a low level of calciotropic hormones.

Methods

Removal of thyroid and parathyroid glands (TPTx) in rats resulted in a decreased level of calciotropic hormones and subsequent bone loss assessed by micro computed tomography (micro-CT) and measurement of serum bone formation and resorption markers, including osteocalcin, C-telopeptide, osteoprotegerin and receptor activator of nuclear factor kappa-B ligand. Results were complemented with in vitro studies on osteoblast and osteoclast activity by both BMP-2 and -7. The doses used were calculated from published pharmacodynamic studies and bioavailability results from preclinical BMP-2 and -7 studies.

Results

TPTx resulted in bone loss, which was restored by systemic administration of 10–70 μg/kg of BMP-2 and 10–250 μg/kg of BMP-7. BMP-2 showed a higher capacity for enhancing trabecular microarchitecture, whereas BMP-7 augmented trabecular thickness. In vitro experiments revealed that BMP-2 and -7 when uncoupled increased the number and activity of both osteoblasts and osteoclasts.

Conclusions

Surprisingly, both BMP-2 and -7 showed an increased bone volume in an in vivo environment of low calciotropic hormones. Locally administered BMP-2 and -7 from bone devices might become partially available in circulation but will not mediate systemic bone loss.     

Using virtual patients to improve cardiac examination competency in medical students

BACKGROUND: Cardiac examination (CE) skills are in decline. Most prior studies employed audio recordings, evaluating only one aspect of CE (i.e., auscultation) that precluded correlation with visible observations. To address these deficiencies, we developed a curriculum using virtual patient examinations (VPEs); bedside recordings of patients with visible and audible cardiovascular findings presented as interactive multimedia. HYPOTHESIS: The purpose of this study was to evaluate whether VPEs improve CE skills, and whether any improvements are retained. We assessed CE competency overall and in 4 categories: inspection, auscultation, knowledge, and integration of audio and visual skills. METHODS: Students (n = 24) undergoing the 8-wk Internal Medicine (IM) clerkship rotation and receiving supervised instruction with VPEs (intervention group) were compared with students (n = 58) undergoing IM clerkship rotation without supplemental CE instruction (control group). The groups were tested at the beginning and the end of their rotations. RESULTS: The Intervention group improved significantly in overall mean scores: from 58.7 to 73.5 (p = 0.0001). The Control group did not improve: from 60.1 to 59.5 (p = 0.788). The Intervention group improved inspection, auscultation, and knowledge (all p 相似文献   

Sex-specific differences of craniofacial traits in Croatia: the impact of environment in a small geographic area

BACKGROUND: Craniometric variation in humans reflects different genetic and environmental influences. Long-term climatic adaptation is less likely to show an impact on size and shape variation in a small local area than at the global level. Aim: The aim of this work was to assess the contribution of the particular environmental factors to body height and craniofacial variability in a small geographic area of Croatia. SUBJECTS AND METHODS: A total of 632 subjects, aged 18-21, participated in the survey. Body height, head length, head breadth, head height, head circumference, cephalic index, morphological face height, face breadth, and facial index were analysed regarding geographic, climatic and dietary conditions in different regions of the country, and correlated with the specific climatic variables (cumulative multiyear sunshine duration, cumulative multiyear average precipitation, multiyear average air temperatures) and calcium concentrations in drinking water. Significant differences between groups classified according to geographic, climatic or dietary affiliation, and the impact of the environmental predictors on the variation in the investigated traits were assessed using multiple forward stepwise regression analyses. RESULTS: Higher body height measures in both sexes were significantly correlated with Mediterranean diet type. Mediterranean diet type also contributed to higher head length and head circumference measures in females. Cephalic index values correlated to geographic regions in both sexes, showing an increase from southern to eastern Croatia. In the same direction, head length significantly decreased in males and head breadth increased in females. Mediterranean climate was associated with higher and narrower faces in females. The analysis of the particular climatic variables did not reveal a significant influence on body height in either sex. Concurrently, climatic features influenced all craniofacial traits in females and only head length and facial index in males. Mediterranean climate, characterized by higher average sunshine duration, higher average precipitation and higher average air temperatures, was associated with longer, higher and narrower skulls, higher head circumference, lower cephalic index, and higher and narrower faces (lower facial index). Calcium concentrations in drinking water did not correlate significantly with any dependent variable. CONCLUSION: A significant effect of environmental factors on body height and craniofacial variability was found in Croatian young adult population. This effect was more pronounced in females, revealing sex-specific craniofacial differentiation. However, the impact of environment was low and may explain only 1.0-7.32% variation of the investigated traits.     

K-ras and Dpc4 mutations in chronic pancreatitis: case series

Aim

To assess whether alterations in the K-ras, p53, and DPC4 genes are present in pancreatitis, a potential precancerous condition that can progress to pancreatic adenocarcinoma. To investigate the alterations occurring at hot spots of K-ras (exon 1), p53 (exons 5 and 7), and DPC4 (exons 8, 10 and 11).

Methods

In 10 patients with acute and 22 with chronic pancreatitis, without pancreatic intraepithelial neoplasia (PanIN), DNA was isolated from paraffin embedded tissue samples. The extracted DNA was analyzed by polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) analysis, single-strand conformation polymorphism (SSCP) analysis, and DNA sequencing.

Results

In acute pancreatitis samples no mutations were found in any of the investigated genes. In 7 out of 22 samples of chronic pancreatitis nucleotide substitution at exon 1 of K-ras (five at codon 12 and two at codon 13) were found. No mutations in p53 (exons 5 and 7) were detected. Two samples had nucleotide substitutions at exons 8 and 11 of DPC4, introducing STOP signal and change in the amino acid sequence, respectively. One chronic pancreatitis sample displayed simultaneous mutations in K-ras (exon 1, codon 12) and DPC4 (exon 8, codon 358).

Conclusion

Mutations of K-ras and Dpc4 genes can accumulate already in non-malignant, inflammatory pancreatic tissue, suggesting its applicability in monitoring of further destruction of pancreatic tissue and progression into malignancy.Pancreatitis is an inflammatory disease of the pancreas, characterized by the progressive tissue destruction, leading both to exocrine and endocrine insufficiency. Acute pancreatitis can manifest as a benign condition with minimal abdominal pain and hyperamylasemia, or can have a fulminate course due to the development of infective pancreatic necrosis and multisystem organ failure (1).On the other hand, chronic pancreatitis is characterized by the progressive destruction of pancreatic parenchyma and its replacement by variable amounts of fibrous tissue (2). A characteristic histological profile displays a fibrosis in association with the foci of inflammatory cells, the regions of acinar cell degradation, and the areas of ductal cell proliferation (3).Diagnosing pancreatitis at an early stage is a clinical challenge, because the frequency of pancreatic ductal adenocarcinoma in patients with pre-existing chronic pancreatitis is significantly higher than in the general population. It has been shown that chronic pancreatitis increases the risk of developing pancreatic cancer 10 to 20 times (4). Progression model of pancreatic ductal adenocarcinoma at the molecular level shows that it always harbors K-ras, p53, and p16 alterations, and also mutations in the DPC4 (deleted in pancreatic cancer gene, locus 4) (5-7). In contrast to pancreatic cancer, the molecular mechanism responsible for acute and chronic pancreatitis is poorly understood. Some improvement has been made with the discovery of the genetic nature of hereditary pancreatitis (8,9). The studies of molecular alterations occurring in acute and chronic pancreatitis have mainly included the investigation of alterations in cationic trypsinogen gene, K-ras, p53, or cystic fibrosis transmembrane conductance regulator gene (CFTR) (8,10-14).Since pancreatitis represents a precancerous state in a considerable number of cases, we assumed that the alterations in more than one gene could be associated with the genetic basis of the disease and the genes involved in the development of pancreatic adenocarcinoma could be mutated already in pancreatitis. Therefore, in collected tissue samples of acute and chronic pancreatitis, we investigated the alterations occurring at hot spots of three different genes (K-ras, p53, DPC4) which are members of different signaling pathways.     

Plasma interleukin-8 as a potential predictor of mortality in adult patients with severe traumatic brain injury

Because of complex pathophysiology and severe consequences, traumatic brain injuries (TBI) are an important medical problem. Pathophysiology of TBI includes local and systemic stress response, in which interleukin-8 (IL-8) is considered as a key mediator of neuroinflammation. However, prognostic relevance of IL-8 measurement in adult patients with severe TBI is not certain. Therefore, IL-8 was determined in blood samples from central venous and jugular bulb catheter and in cerebrospinal fluid of twenty patients with isolated TBI at admission to Intensive Care Unit. None of the patients had history of stroke, dementia, autoimmune diseases, acute infection or medication with anti-inflammatory drugs. Ten patients died due to traumatic brain injury, while the other ten recovered well. While there was no significant difference of IL-8 levels in cerebrospinal fluid between survivors and nonsurvivors, central venous plasma level of IL-8 was significantly lower in survivors (71.00 +/- 14.17 pg/ml), than in nonsurvivors (111.26 +/- 16.9 pg/ml). Receiver Operating Characteristic (ROC) analysis revealed significant prognostic value for IL-8 in the blood as well as for the age of patients, Glasgow Coma Scale (GCS) and Acute Physiologic and Chronic Health Evaluation (APACHE II). These findings suggest that the central venous plasma values of IL-8 at admission might be an early predictive marker in patients with severe TBI, comparative to standard clinical prognostic markers such as APACHE II and GCS.     

Naphthalanotherapy reduces angiogenetic factor in psoriatic lesion

Immunohistochemical analysis (cell immunophenotyping before and after 3 weeks of treatment with naphthalan oil) was performed on biopsy specimens from 10 patients with psoriasis vulgaris. To determine the angiogenetic factor in psoriatic lesions, immunohistochemical staining on 3-microm paraffin block sections was performed by using monoclonal anti-factor VIII antibody. There was a significant difference in the mean number of new blood vessels before and after the therapy (15.1 vs. 6.7). It seems that naphthalan therapy reduces number of new blood vessels and has effect on neovascularization in patients with psoriasis.     

Transcatheter aortic valve implantation: patient selection and procedural considerations

Transcatheter aortic valve implantation has risen exponentially as a treatment modality for severe aortic stenosis in patients considered to be at high risk for or inoperable by conventional surgical aortic valve replacement. It has shown both survival and quality of life benefit in a randomized comparison to conservative (palliative) therapy in nonoperative candidates. Fundamental to its success is appropriate patient selection and a rigorous attention to procedural steps. In this article we will discuss the key issues pertaining to each of these factors.