Anticancer Activity of Tubulosine through Suppression of Interleukin-6-Induced Janus Kinase 2/Signal Transducer and Activation of Transcription 3 Signaling / 한국유방암학회지

PURPOSE: The chemical structure of tubulosine has been known since the mid-1960s. However, little is known about its biological and pharmacological functions. The aim of this study was to investigate the novel functions of tubulosine in cancer treatment, specifically in breast cancer. METHODS: An Unpaired (Upd)-induced Drosophila cell line and interleukin (IL)-6-stimulated human breast cancer cell lines were used to investigate the biological and pharmacological activities of tubulosine in vitro. To investigate the activities of tubulosine, we performed molecular and cellular experiments such as Western blot and reverse transcription polymerase chain reaction analyses, immunoprecipitation and terminal deoxynucleotidyl transferase dUTP nick end labeling assays, and immunofluorescence staining using breast cancer cell lines. RESULTS: Tubulosine exhibited anticancer activity in IL-6-stimulated human breast cancer cells. Moreover, tubulosine reduced the tyrosine phosphorylation level and transcriptional activity of signal transducer and activator of transcription (STAT) protein at 92E in Upd-induced Drosophila cells. Additionally, tubulosine suppressed IL-6-induced Janus kinase 2 (JAK2)/STAT3 signaling, resulting in decreased viability and induction of apoptotic cell death in breast cancer cells. Interestingly, inhibition of IL-6-induced JAK2/STAT3 signaling by tubulosine was associated with the blocking of IL-6 receptor (IL-6R) and glycoprotein 130 (gp130) binding. CONCLUSION: Tubulosine exhibits anticancer activity through functional inhibition of IL-6-induced JAK2/STAT3 signaling by targeting IL-6Rα/gp130 binding in breast cancer cells. These findings suggest that tubulosine may hold promise for the treatment of inflammation-associated cancers, including breast cancer.

Chronic Non-Social Stress Affects Depressive Behaviors But Not Anxiety in Mice

The etiology of most psychiatric disorders is still incompletely understood. However, growing evidence suggests that stress is a potent environmental risk factor for depression and anxiety. In rodents, various stress paradigms have been developed, but psychosocial stress paradigms have received more attention than non-social stress paradigms because psychosocial stress is more prevalent in humans. Interestingly, some recent studies suggest that chronic psychosocial stress and social isolation affects mainly anxiety-related behaviors in mice. However, it is unclear whether chronic non-social stress induces both depression- and anxiety-related phenotypes or induces one specific phenotype in mice. In the present study, we examined the behavioral consequences of three chronic non-social stress paradigms: chronic predictable (restraint) stress (CPS), chronic unpredictable stress (CUS), and repeated corticosterone-HBC complex injection (RCI). Each of the three paradigms induced mild to severe depression/despair-like behaviors in mice and resulted in increased immobility in a tail suspension test. However, anxiety-related phenotypes, thigmotaxis and explorative behaviors, were not changed by the three paradigms. These results suggest that depression- and anxiety-related phenotypes can be dissociated in mouse stress models and that social and non-social stressors might affect brain circuits and behaviors differently.

Anti-inflammatory activity of compounds isolated from Astragalus sinicus L. in cytokine-induced keratinocytes and skin

Inflammation is a part of the complex biological responses of a tissue to injury that protect the organ by removing injurious stimuli and initiating the healing process, and is considered as a mechanism of innate immunity. To identify biologically active compounds against pathogenic inflammatory and immune responses, we fractionated water, aqueous methanol and n-hexane layers from nine kinds of leguminosae and examined anti-inflammatory activity of the fractions in human keratinocytes and mouse skin. Among the fractions, rf3 and rf4, isolated from the aqueous methanol layer of Astragalus sinicus L., exhibited the strongest reactive oxygen species (ROS)-scavenging and anti-inflammatory activities as measured by inhibition of the intracellular ROS production, nuclear factor-kappaB (NF-kappaB), janus kinase (JAK)/signal transducer and activator of transcription (STAT), and phosphatidylinositol 3-kinase/Akt signaling in cytokine-stimulated human keratinocytes, as well as by effects on T-cell differentiation in mouse CD4+ T cells. In addition, topical application of rf3 and rf4 suppressed the progression of psoriasis-like dermatitis and expression of pro-inflammatory mediators in interleukin (IL)-23-injected mouse ears. Our results suggest that Astragalus sinicus L. may ameliorate chronic inflammatory skin diseases due to its antioxidant and anti-inflammatory activities via regulation of the intracellular ROS production, NF-kappaB, JAK/STAT and PI3/Akt signaling cascades as well as immune responses, and these results are the first report that Astragalus sinicus L. exhibits pharmacological activity.

Cementless Bipolar Hemiarthroplasty for Unstable Intertrochanteic Fractures in the Elderly / 대한정형외과학회잡지

PURPOSE: To evaluate the positive effects and problems through clinical and radiological results of cementless bipolar hemiarthroplasty for intertrochanteric fractures above type A2 in the elderly. MATERIALS AND METHODS: From December 2006 to June 2009, 54 bipolar hemiarthroplasties were performed in 54 patients in our hospital. The mean age was 78.8 (67-93) years. Of these cases, 13 were male and 41 were female. The fractures were of type A2.1 in 17 cases, type A2.2 in 23 cases and type A2.3 in 14 cases. There was no walking limitation in 45 patients, but 4 of the remaining 9 patients had walking limitations and used walking aids at their residence. A posterolateral approach as well as cementless femoral stems was used in all the patients. Clinical results were evaluated according to operation time, amount of bleeding, time to resume walking, duration of hospital stay, recovery of walking ability, and complications. Prostheses loss was evaluated on the follow-up radiographs. RESULTS: Twenty-two of 39 patients who had medical co-morbidity had more than two medical co-morbidities. Operations were performed at a mean time of 5.5 days after the fracture. The mean operation time was 95 minutes. The average total amount of bleeding was of 715 cc. Patients began walking at an average of 5.9 days after operation and the average duration of hospital stay was 19.2 days. Sixteen patients (29.6%) died at an average period of 1.6 years after their operation. At the time of discharge, 32 patients (59%) had recovered walking ability, but at the last follow-up compared to the pre-injury status, the recovery rate of walking had decreased to 46% (25 patients). Complications included a deep infection in one case, dislocation in 2 cases and hematoma in 2 cases. The cause of revision was deep infection. There were no revisions due to prosthesis loosening. CONCLUSION: Cementless bipolar hemiarthroplasty for intertrochanteric fractures in the elderly had some problems due to the prolonged operation time and increased amount of bleeding, but it also had advantages including the early return to walking after the operation and decreased hospital stay. It is one of the treatment options for the elderly with unstable intertrochanteric fractures.

Total Hip Arthroplasty Using Two-Incision Technique

BACKGROUND: To evaluate the effectiveness of minimally invasive surgery total hip arthroplasty (THA) using the two-incision technique as described by Mears. METHODS: From January 2003 to December 2006, sixty-four patients underwent total hip arthroplasty using the one-incision (group I) and two-incision (group II) technique by one surgeon. There were 34 hips in group I and 30 hips in group II. There was no difference in age, gender, and causes of THA between the two groups. We evaluated the operation time, bleeding amount, incision length, ambulation, hospital stay, and complications between the two groups. RESULTS: There was no difference in the bleeding amount between the two groups. Operation time was longer in the two-incision group than in the one-incision group. Operation time of the two-incision technique could be reduced after 15 cases. Patients started ambulation after surgery earlier in group II than group I, and the hospital stay was shorter in group II than in group I. There was no difference in clinical results between the two groups. There was no difference in component position of the acetabular cup and femoral stem between the two groups. Intraoperative periprosthetic fracture occurred in four cases (13.3%) in group II. CONCLUSIONS: Two-incision THA has the advantage of rapid recovery and shorter hospital stay. However, longer operation time and a high complication rate compared to one-incision are problems that need to be solved in the two-incision technique.

Benzoxathiol derivative BOT-4-one suppresses L540 lymphoma cell survival and proliferation via inhibition of JAK3/STAT3 signaling

Persistently activated JAK/STAT3 signaling pathway plays a pivotal role in various human cancers including major carcinomas and hematologic tumors, and is implicated in cancer cell survival and proliferation. Therefore, inhibition of JAK/STAT3 signaling may be a clinical application in cancer therapy. Here, we report that 2-cyclohexylimino-6-methyl-6,7-dihydro-5H-benzo [1,3]oxathiol-4-one (BOT-4-one), a small molecule inhibitor of JAK/STAT3 signaling, induces apoptosis through inhibition of STAT3 activation. BOT-4-one suppressed cytokine (upd)-induced tyrosine phosphorylation and transcriptional activity of STAT92E, the sole Drosophila STAT homolog. Consequently, BOT-4-one significantly inhibited STAT3 tyrosine phosphorylation and expression of STAT3 downstream target gene SOCS3 in various human cancer cell lines, and its effect was more potent in JAK3-activated Hodgkin's lymphoma cell line than in JAK2-activated breast cancer and prostate cancer cell lines. In addition, BOT-4-one-treated Hodgkin's lymphoma cells showed decreased cell survival and proliferation by inducing apoptosis through down-regulation of STAT3 downstream target anti-apoptotic gene expression. These results suggest that BOT-4-one is a novel small molecule inhibitor of JAK3/STAT3 signaling and may have therapeutic potential in the treatment of human cancers harboring aberrant JAK3/STAT3 signaling, specifically Hodgkin's lymphoma.

Two-Incision Bipolar Hemiarthroplasty for Treating Femoral Neck Fracture: Analysis of 15 Cases

INTRODUCTION: We wanted to evaluate the efficacy of bipolar hemiarthroplasty using Berger's two-incision surgical technique for patients with muscular weakness around the hip joint and/or a high risk of dislocation. MATERIALS AND METHODS: We performed bipolar hemiarthroplasty for 15 femoral neck fractures using Berger's two-incision technique between December 2005 and July 2007. The mean age of the patients was 75.2 years old. Four of them had difficulty in walking due to the sequalae after stroke and five have been treated for psychoneurologic disorders. We investigated the operation time, the length of the anterior and posterior incisions, the amount of bleeding, the time untill walking after the operation, the total hospital stay, the recovery to activities of daily living and the complications such as dislocation. RESULTS: The mean operating time was 93 minutes. The average anterior and posterior skin incision length was 6.4 cm and 7.2 cm, respectively. The mean amount of bleeding was 420 cc at the time of surgery and 230 cc postoperatively through a drain. The patients started walking at a mean of 3.3 (1 to 5) days after the operation and the mean hospitalization was 24.3 days. Fourteen patients went back to their pre-injured activities of daily living, except one case with an intraoperative periprosthetic fracture. As for complications, two cases (13.3%) of femoral fracture were intraoperatively observed and one case of skin necrosis on the anterior incision site occurred. There were not any cases of dislocation or infection. CONCLUSION: Two-incision bipolar hemiarthroplasty had advantages for rehabilitation in elderly patients who have a high risk of dislocation, as well as in the patients with muscle weakness. But the operation took a long time and it had a high complication rate.

A Study on Reference Values from Health Checkup Data of Korea Association of Health Promotion by Indirect Method: A Study on Standardization of Reference Values among Laboratories of Korea Association of Health Promotion II / 임상검사와정도관리

BACKGROUND: The reference values of laboratory should review regularly to maintain good quality of practice. This is the second report of studies on reference values of Korea Association of Health Promotion (KAHP). The first one was reported in 2002 in Journal of Laboratory Medicine and Quality Assurance in Korea. The aim of this study is to know the changes of reference values for the past five years. METHODS: The way to analyze the data this time was essentially the same as the previous one (Indirect Method). The data from January to December, 2007 were collected. They totalled 5,133,327 test results from approximately 140 thousands individuals who visited for health checkup. The data were statistically analyzed with Minitab version 15.1.20.0 for Gaussian distribution using Anderson-Darling test. The trimming process repeated for the outliers, the results lying outside of +/-3SD, and as much as four times in certain test items, though, none of the tests showed Gaussian distribution. Subsequently, the reference values of most tests were defined in the ranges from the point of lowest 2.5% to the point of highest 97.5% and others were those, below 95 percentiles according to CLSI C28-A3 guideline. RESULTS: The reference ranges of 56 test items were either set as before or adjusted with new values, and compared. CONCLUSIONS: Comparing to the previous reference values (2002), the tests for liver function showed the lower upper values and the tests for diabetes and lipids showed higher upper values. Others were changed minimally with no significance.

The Effectiveness of Bone Mineral Density as Supplementary Tool for Evaluation of the Osteogenic Potential in Patients with Spinal Fusion

STUDY DESIGN: Retrospective study. PURPOSE: This study was designed to determine the effectiveness of bone mineral density measurement as a supplementary tool for evaluation of osteogenic potential in patients with spinal fusion. To this end, we correlated bone mineral density (BMD) with osteogenic potential from cultured mesenchymal stem cells (MSCs). OVERVIEW OF LITERATURE: Many studies have correlated osteogenic potential of in vitro cultured MSCs with aging or osteoporosis. METHODS: We studied twenty-five individuals with harvested bone marrow from the ilium during lumbar spinal surgery. The BMD of the femoral neck was measured using dual energy X-ray absorptiometry prior to bone marrow aspiration, and the osteoporotic group was classified as those with T-scores below-2.5. After MSCs were isolated from bone marrow, in vitro induction of osteogenesis was performed. We analyzed the patient's osteogenic potential from cultured MSCs such as mineral deposition stain, bone alkaline phosphatase (ALP) activity and osteoblast-specific gene expression in RT-PCR. RESULTS: On mineral staining, the osteoporotic group had a scanty matrix mineral deposition in contrast to the non-osteoporotic group. The expression of osteocalcin in the osteoporotic group was 1.5 to 3 times less than in the non-osteoporotic group. At the 3rd week after the induction of osteogenesis, the activity of ALP of cultured MSCs in the osteoporotic group was lower than in the control group (mean, 45+/-19 u/L, in osteoporotic group vs 136+/-7 u/L in non-osteoporotic), and there was a statistically significant and positive correlation between BMD & ALP (r=0.487, p=0.013). CONCLUSIONS: There is a positive correlation between BMD and osteogenic potential derived from MSCs. The measurement of BMD can provide supplementary data for evaluating osteogenic potential clinically.

Treatment of the Femoral Shaft Nonunion Occurredafter Intramedullary Nailing / 대한정형외과학회잡지

PURPOSE: To evaluate of efficacy of the treatment options for a femoral shaft nonunion occurring after intramedullary nailing. MATERIAL AND METHODS: Thirty-one patients with nonunion of a femoral shaft fracture, who had been treated with interlocking intramedullary nailing from January 1996 to December 2000, were examined. Twenty-six had oligotrophic nonunion and five were hypervascular. Forty-five procedures were performed for 31 nonunions; bone grafting for 14, exchange nailing for 13, plate augmentation and bone grafting for 14 and dynamization for 4 cases. RESULTS: The success rate after a single procedure was only 58%. The four dynamization cases failed to unite. Seven of the 13 (54%) nonunion cases treated with nail exchange healed satisfactorily. All cases treated with plate augmentation and bone grafting achieved successful union. The mean period from fracture to union was 20 months. CONCLUSION: Exchange nailing is not always a reliable procedure for treating nonunion of a femoral shaft fracture. Plate augmentation and bone grafting were found to be a successful mode of therapy for the femoral shaft nonunion without complications.

Establishment of a Multipotent Neural Stem Cell Line from the Adult Mouse Cerebrum / 대한척추외과학회지

STUDY DESIGN: Establishment of a multipotent neural stem cell line from the adult mouse cerebrum. OBJECTIVES: To establish a daughter cell line, B2A1, from B2 cells through the limiting dilution method, and to determine if the cells have the characteristics of neural stem cell (NSCs) using immunocytochemistry and RT-PCR methods. SUMMARY AND LITERATURE REVIEW: In the development of NSCs, differentiated organ or tissue-derived multipotent stem cells have attracted considerable interest because of the lack of ethical issues. Previously, a glial precursor cell line (B2 cells) was generated from the primary cultures of oligodendrocytes/ astrocytes in an adult BALB/c mouse brain. These cells exhibited the cell-type specific markers for immature neuroectodermal cells, astrocytes, and oligodendrocytes in serum-contained media. MATERIALS AND METHODS: The primary cultures of oligodendrocytes/astrocytes were established from the whole brains of 12 to 16-week-old BALB/c mice from either gender. After 6 months with 25 serial passages, the culture consisted of a morphologically homogeneous cell population, which was designated as B2 cells. A subclone, B2A1, was isolated from B2 cells through two consecutive limiting dilutions. RESULTS: More than 90% of B2A1 cells showed immunopositivity for nestin, a specific marker for NSC. The cells also showed immunopositivity for the neuronal, astrocytic and oligodendroglial markers. These cells expressed the genotypic mRNA messages for both neural progenitor cells and differentiated neuronoglial cells. These positive immunocytochemical reactions and mRNA messages for neuronoglial cells varied according to the extrinsic growth factors used. However, the treatment of extrinsic growth factors did not produce any significant differences in the nestin-immunopositive cells. CONCLUSIONS: B2A1 cells have the immunocytochemical and cytogenetic properties of NSCs, and the capacity to differentiate into neuronoglial cells.

Involvements of Oxidative Stress in beta-amyloid Peptide: induced Cytotoxicity in PC12 Cells

BACKGROUND: Alzheimer's disease is a neurodegenerative disorder characterized by the extracellular deposition of beta-amyloid peptide(Abeta) in the brain, presumed to play a pathogenic role. However, the precise molecular mechanisms of its neurotoxicity are not fully understood. METHODS: Abeta-mediated cytotoxicity in neuronal cell lines (PC12, SH-SY5Y, IMR32, and U87) was measured by an MTT assay. NF-kappaB activation by Abetawas examined by a luciferase assay and apoptosis induced by Abetawas measured by cytoplasmic DNA fragmentations. RESULTS: Abetacytotoxicity in the tested cell lines was more prominent in the absence of serum than in the presence of serum in culture media. PC12 cells showed the highest sensitivity to Abetacytotoxicity among the cell lines. The Abeta(25-35) cytotoxicity in PC12 cells was increased in a dose-dependent manner. For convincing oxidative stress involved in Abetacytotoxicity, antioxidants such as DTT, GSH, vitamin C, or NAC were pretreated. GSH protected PC12 cells from Abetacytotoxicity, but DTT or NAC did not. Abeta (25-35) treatment to PC12 cells increased the NF-kappaB activity in a dose-dependent manner. Cytoplasmic DNA fragmentations, one of the apoptotic indicators, were increased at lower concentrations of Abeta(25-35) from 0.01 to 0.1 microM, however, dose-dependent increments of DNA fragmentations were not observed at higher concentrations from 1 to 10 microM. CONCLUSIONS: From these results, Abeta-induced cytotoxicity in PC12 cells might be mediated by oxidative stress.