OBJECTIVE: Recently, microdissection of tissue sections has been used increasingly for the isolation of morphologically identified homogeneous cell populations, thus overcoming the obstacle of tissue complexity for the analysis cell-specific expression of macromolecules. The aim of the present study was to establish the minimal conditions required for the RNA extraction and amplification from the cells captured by the laser captured microdissection. METHODS: Mouse ovaries were fixed and cut into serial sections (7 micrometer thickness). Oocytes were captured by laser captured microdissection (LCM) method by using PixCell IITM system. The frozen sections were fixed in 70% ethanol and stained with hematoxylin and eosin, while the paraffin sections were stained with Multiple stain. Sections were dehydrated in graded alcohols followed by xylene and air-dried for 20 min prior to LCM. All reactions were performed in ribonuclease free solutions to prevent RNA degradation. After LCM, total RNA extraction from the captured oocytes was performed using the guanidinium isothiocyanate (GITC) solution, and subsequently evaluated by reverse transcriptase -polymerase chain reaction (RT-PCR) for glyceraldehyde-3-phosphate-dehydrogenase (GAPDH). RESULTS: With the frozen sections, detection of the GAPDH mRNA expression in the number of captured 25 oocytes were not repeatable, but the expression was always detectable from 50 oocytes. With 25 oocytes, at least 27 PCR cycles were required, whereas with 50 oocytes, 21 cycles were enough to detect GAPDH expression. Amount of the primary cDNA required for RT-PCR was reduced down to at least 0.25 microl with 50 oocytes, thus the resting 19.75 microl cDNA can be used for the testing other interested gene expression. Tissue-to-slide, tissue-to-tissue forces were very high in the paraffin sections, thus the greater number of cell procurement was required than the frozen sections. CONCLUSION: We have described a method for analyzing gene expression at the RNA level with the homogeneously microdissected cells from the small amount of tissues with complexity. We found that LCM coupled with RT-PCR could detect housekeeping gene expression in 50 oocytes captured. This technique can be easily applied for the study of gene expression with the small amount of tissues.
Alcohols ; Animals ; DNA, Complementary ; Eosine Yellowish-(YS) ; Ethanol ; Female ; Frozen Sections ; Gene Expression* ; Genes, Essential ; Guanidine ; Hematoxylin ; Mice ; Microdissection* ; Oocytes* ; Ovary ; Paraffin ; Polymerase Chain Reaction ; Ribonucleases ; RNA Stability ; RNA* ; RNA, Messenger ; RNA-Directed DNA Polymerase ; Xylenes

PURPOSE: Tumor invasion and metastasis are multistep phenomina, involving proteolytic degradation of extracellular matrix (ECM) and alteration of cell adhesion. It is another oncologic therapeutic strategy to block tumor invasion and metastasis through analyzing the molecules which are involved in these processes. We examined the expressions of some of matrix metalloproteinases (MMPs) in colorectal cancer and analyze the correlation with clinical factors and survival. METHODS: Fifty-five patients with colorectal cancer who underwent surgical resection were included. The expressions of the MMP-2, -7, -9, and -12 in tumor tissue and normal mucosa using RT-PCR technique was carried out. We evaluated and analyzed the correlation among these molecules, clinical characteristics, and survival. RESULTS: Expressions of MMP-7, -9, and -12 were significantly higher in tumor tissue than in normal mucosa (P=0.00). The expressions of MMP-2 between cancer and normal mucosa had no significant difference but it had a significant difference according to the lymph node (LN) invasion (P=0.03) in tumor tissues. Three-year overall survival was worse in patient with high expression of MMP-2 compared to low expression. CONCLUSION: The expressions of MMP-7, -9, -12 in tumor tissue were higher than in normal mucosa and MMP-2 expression of tumor tissues had a significant difference according to LN invasion. MMP-2 overexpression seems to be a prognostic factor for 3-yr overall survival. But, large scale study with long term survival analysis will be needed for the prognostic significance of MMPs.
Cell Adhesion ; Colorectal Neoplasms ; Extracellular Matrix ; Humans ; Lymph Nodes ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinases ; Mucous Membrane ; Neoplasm Metastasis

No abstract available.
Child ; Humans ; Kasabach-Merritt Syndrome* ; Kidney Failure, Chronic ; Peritoneal Dialysis, Continuous Ambulatory

No abstract available.
Child ; Humans ; Kasabach-Merritt Syndrome* ; Kidney Failure, Chronic ; Peritoneal Dialysis, Continuous Ambulatory

BACKGROUND: Quercetin, found abundantly in onion peel, has been known to have anticholesterol, antithrombotic and insulin-sensitizing properties. Here, we investigated the effect of quercetin-rich onion peel extract (OPE) on reactive oxygen species (ROS) production and antioxidative defense in obese woman. METHODS: This study was randomized, double-blind, placebo controlled study. Thirty-seven healthy obese participants were randomly assigned that eighteen subjects received red soft capsuled OPE (100 mg/d, 50 mg bis in die), while the other nineteen subjects received same capsuled placebo for 12 weeks. ROS production and superoxide dismutase (SOD) activity in plasma were determined by using ROS and SOD assay kits, respectively. RESULTS: Baseline characteristics of anthropometric indicators and blood metabolic profiles were not significantly different between the two groups. Compared with baseline values, OPE consumption significantly reduced waist and hip circumference. Plasma ROS level and SOD activity were decreased in both placebo and OPE groups compared with baseline values. However, plasma ROS level in OPE group was significantly lower than in placebo group while plasma SOD activity in OPE group was significantly higher than in placebo group after 12 weeks of consumption. CONCLUSIONS: These findings indicate that OPE consumption may exert antioxidative effect by preventing the decrease of SOD activity as well as the production of ROS in obese women.
Female ; Hip ; Humans ; Metabolome ; Onions* ; Plasma ; Quercetin ; Reactive Oxygen Species ; Superoxide Dismutase

Quercetin, found abundantly in onion peel, has been known to have antioxidant and anti-obesity effects and improves endothelial function. The purpose of this study was to evaluate the effects of a quercetin-rich onion peel extract (OPE) on the inflammatory mediators in overweight and obese women. This study was a randomized double-blind, placebo-controlled study. Thirty-seven healthy overweight and obese women were randomly assigned to two groups, and one group was given a soft capsuled OPE (100 mg quercetin/day, n = 18) and the other group a same capsuled placebo (n = 19) for 12 weeks. Fat mass was measured by bioimpendance method at baseline and after 12 weeks of intervention. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured with colorimetric assay kits. The concentrations of leptin, adiponectin, visfatin, tumor necrosis factor (TNF)-α and interleukin (IL)-4 in plasma were determined by using enzyme-linked immunosorbent assay kits. Baseline characteristics of anthropometric indicators and blood metabolic profiles were not significantly different between placebo and OPE groups. Compared with baseline value, both placebo and OPE supplementation significantly decreased the percent of body fat mass and induced plasma adiponectin levels while ALT and AST activities as well as leptin, visfatin, TNF-α, and IL-4 levels in plasma were not significantly different between two groups after 12 weeks of the supplementation. These findings suggest that 12-week supplementation of OPE do not affect modulators of systemic inflammation in overweight and obese women.
Adiponectin ; Adipose Tissue ; Alanine Transaminase ; Aspartate Aminotransferases ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Inflammation ; Interleukin-4 ; Interleukins ; Leptin ; Metabolome ; Methods ; Nicotinamide Phosphoribosyltransferase ; Obesity ; Onions* ; Overweight* ; Plasma ; Quercetin ; Tumor Necrosis Factor-alpha

Purpose: This study is a hypothetical model of the effect of the leader-member exchange relationship on psychological ownership, job engagement, and turnover intention. Methods: Data collection was conducted for general nurses who have worked for at least 6 months in a university hospital. The collected data were analyzed using the SPSS/WIN 25.0, and AMOS 18.0 program was used to conduct confirmatory factor analysis as well as goodness of fit verification and hypothesis testing for the hypothetical model. Results: The leader-member exchange had a significant, static effect on psychological ownership. The leader-member exchange relationship did not have a significant effect on job engagement and psychological ownership had a significant, static effect on job engagement.The leader-member exchange relationship did not have a significant effect on turnover intention and psychological ownership and job engagement had significant, negative effects on turnover intention. Conclusion Intervention and the development of programs to increase the psychological ownership and job engagement of members are proposed to improve the leader-member exchange relationship through leadership education and training and reduce the turnover intention of nurses.

OBJECTIVES: To access the effectiveness of radiofrequency myolysis (RFM) in women with midline dysmenorrhea. METHODS: We designed RFM in two ways laparoscopic RFM (LRFM), vaginal ultrasound-guided RFM (URFM). One hundred and thirty-two patients were in the LRFM group and, 140 patients were in the URFM group. RESULTS: Upon receipt of surgery, both the LRFM and the URFM groups demonstrated a significant decrease (P < 0.001) in the mean pain score when compared to those before and after surgery. CONCLUSION: The RF uterine myolysis procedure provides an alternative for those patients who suffer from intractable midline dysmenorrhea. LRFM is an alternative choice because it is relatively safe and, simple to perform and moreover, it is satisfactory. LRFM appears to increasingly succeed in the treatment of midline dysmenorrhea.
Dysmenorrhea* ; Female ; Humans ; Laparoscopy ; Leiomyoma ; Ultrasonography

PURPOSE: To evaluate Metal Artifact Reduction for Orthopedic Implants (O-MAR, Philips Healthcare) technique compared with standard filtered back projection (SFBP) technique on post-operative hip CT regarding image noise reduction and detection of post-operative complications. MATERIALS AND METHODS: Fifty-six hip CT scans after hip replacement with SFBP technique and O-MAR application were retrospectively reviewed. Region of interests (ROIs) were drawn at levels wherein acetabular cup and femoral head were largest at anterior and posterior acetabula, gluteus maximus muscle, subcutaneous fat adjacent to gluteus maximus muscle, and in area adjacent to prosthesis stem wherein lesser trochanter is largest. Hounsfield units (HU) were measured to evaluate artifact quantitatively; mean and standard deviations (SDs) calculated and compared. Periprosthetic complications were evaluated, and visibility rated between two reconstruction techniques; 1-SFBP better, 2–SFBP same as O-MAR, 3–O-MAR better. RESULTS: Average HU was significantly lower in O-MAR at posterior acetabulum, gluteus maximus muscle, and subcutaneous fat (p < 0.05). SD for HU was significantly lower in O-MAR at all ROIs (p < 0.05). Mean visibility of periprosthetic complications was 2.0, so equivalent. CONCLUSION Reconstruction with O-MAR technique in post-operative hip CT scans after hip replacement yielded statistically significant decrease in image noise. However, visibility of periprosthetic complications was not impacted by reconstruction technique.

Pulmonary arterial hypertension is a critical manifestation of systemic sclerosis (SSc) and is a main cause of death. Several treatment modalities for SSc have been identified, with effects that improve quality of life and mortality rates. However, whether these drugs can also normalize pulmonary arterial pressure, remains unclear. Here, we report the case of a woman with diffuse SSc with pulmonary arterial hypertension, who had a functional status equivalent to the New York Heart Association class III. The patient was treated with inhaled iloprost. After six years of inhaled iloprost therapy, echocardiography showed that pulmonary arterial pressure normalized, accompanied by improvement in functional capacity. Inhaled iloprost might not only normalize pulmonary arterial pressure, but also improve the functional status of patients with SSc with pulmonary arterial hypertension.
Arterial Pressure ; Cause of Death ; Echocardiography ; Female ; Heart ; Humans ; Hypertension* ; Hypertension, Pulmonary ; Iloprost* ; Mortality ; Quality of Life ; Scleroderma, Systemic*