Transforming growth factor-j31 (TGF-p1) has potential for therapeutic use in common clinical conditions for which there are no adequate pharmacological agents. However, in vivo studies using TGF-p1 were hindered by high price of this cytokine. As a first step towards large scale purification of TGF-p1, it was purified in a small scale (10 unit platelets) from human platelets by four purification steps: platelet extraction, gel filtration, cation exchange chromatography, and reversed phase high performance liquid chromatography (HPLC). A single protein band with a molecular weight of 25 Kd corresponding to purchased TGF-p1 (R8D Systems) was confirmed by silver staining after SDS-polyacrylamide gel electrophoresis (SDS-PAGE) of eluant from reversed phase HPLC. Recovery (%) of each step was about 50-60%, resulting in the final recovery of 20% based on the detection by a sandwich ELISA. Approximately, 3.7 p,g of purified TGF-p1 was obtained from 18 pg of platelet extracts. This result was confirmed by receptor (TGF-j31 type II) ELISA and bioassay using a mink lung epithelial'cell line (MV1LU). Further, in vitro characterization study showed that purified TGF-p1 inhibits G1/S transition of LPS-activated murine spleen B cells and increases surface IgA expression by the same cell population, which are typical activities of TGF-p1 in B cell differentiation. Taken together, the results from the present study reveals that purified TGF-p1 is fully biologically active and our purification methodology could be usbful to obtain a large scale of recombinant TGF-p1 in the future.
B-Lymphocytes ; Biological Assay ; Blood Platelets ; Cell Cycle ; Cell Differentiation ; Chromatography ; Chromatography, Gel ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Electrophoresis ; Enzyme-Linked Immunosorbent Assay ; Humans* ; Immunoglobulin A ; Lung ; Mink ; Molecular Weight ; Silver Staining ; Spleen ; Transforming Growth Factors*

OBJECTIVESChunghyuldan (CHD), a combinatorial drug that has anti-hyperlipidemic and antiinflammatory activities, has been shown to reduce infarct volume in a focal ischemia-reperfusion rat model. To explore the molecular basis of CHD's neuroprotective effect, we examined whether CHD shows a cell-protective activity and has a regulatory effect on Bax and/or B-cell leukemia/lymphoma 2 (Bcl-2) expression in mouse neuroblastoma 2a (N2a) cells subjected to hypoxia-reoxygenation (H/R).

METHODSIn order to evaluate the effects of CHD on the cytotoxicity induced from hypoxia or H/R condition, lactate dehydrogenase (LDH) assay was performed. To explore whether the suppression of neural damage when pre-treated with CHD is associated with its anti-apoptotic effect, the CHD effect on the expression of Bcl-2 and Bax was analyzed by Western blotting analysis.

RESULTSCytotoxicity of N2a cell line was slightly increased in 42 h hypoxia condition and dramatically increased under the H/R condition. CHD treatment markedly decreased the cytotoxicity in both conditions (P<0.01, P<0.05). H/R markedly increased the expression of the pro-apoptotic protein, Bax, but slightly increased the expression of the anti-apoptotic protein, Bcl-2, compared with the normoxia or hypoxia group. CHD significantly decreased Bax expression (P<0.01) and slightly decreased Bcl-2 expression (P>0.05), resulted in a reduction of Bax/Bcl-2 ratio in N2a cells subjected to H/R.

CONCLUSIONCHD has neuroprotective effect in N2a cells subjected to H/R, which might be derived at least in part from its ability to decrease the expression of the pro-apoptotic protein, Bax.

Animals ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Hypoxia ; prevention & control ; Mice ; Neuroblastoma ; metabolism ; pathology ; Neuroprotective Agents ; pharmacology ; Reperfusion Injury ; prevention & control

Bee stings can cause various skin reactions that usually resolve over several days; however, in some patients the venom acts as an allergen, causing a severe immunological response such as anaphylaxis. Bee stings can also induce chronic inflammation because the barbed stinging apparatus and venom sacs, along with the nerve plexus, can lodge in the skin. Chronic non-specific low back pain is the most common medical problem for which patients seek complementary and alternative medical treatment, including bee venom acupuncture. Bee venom acupuncture involves injecting diluted bee venom into acupoints and is used for arthritis, pain, and rheumatoid diseases. Here we report a 75-year-old man with acute urticaria and skin eruption with eosinophilia and interstitial pneumonia 6 weeks later after receiving 5 times bee venom acupuncture.
Acupuncture Points ; Acupuncture* ; Aged ; Anaphylaxis ; Arthritis ; Bee Venoms ; Bees* ; Bites and Stings ; Eosinophilia ; Humans ; Inflammation ; Low Back Pain ; Lung Diseases, Interstitial ; Skin ; Stevens-Johnson Syndrome ; Urticaria ; Venoms

This study evaluated the microleakage and interfacial gap between enamel and composite resin under the dry and wet condition of the enamel surface. V shaped class 5 cavities were prepared on the occlusal portion of extracted human molars. Samples were divided into three groups: D group (air dry for 10-15 s), BD group (blot dry with moist cotton pellet), and DR group (air dry for 10-15 s and rewet with Aqua-Prep F for 20 s). Cavities were filled using Aelitefil composite resin after applied One-Step. Microleakage was tested by 2% methylene blue dye solution and the data were statistically analysed by Kruskal-Wallis test and Mann-Whitney test. Also Enamel-resin interface was observed under SEM. Group BD showed statistically lower microleakage than group D (p < 0.05), but there was no statistically significant difference between group BD and DR (p > 0.05). At the enamel-resin interface, group D showed the gap of 2 microm thickness, but group BD and DR showed close adaptation. In conclusion, the use of blot dry and rewetting agent (Aqua-Prep F) resulted in decreased microleakage and improved adhesion between enamel and resin when using One-Step.
Dental Enamel* ; Humans ; Methylene Blue ; Molar

INTRODUCTION: There have been no reports of hypersensitivity pneumonitis caused by metal working fluids in Korea, despite their existence in other countries. Here, we report the first such case, along with an assessment of work-relatedness through exposure assessment. CASE REPORT: A 64-year old male patient visited the hospital with dyspnea after metal pipe cutting for about a year. He was diagnosed with hypersensitivity pneumonitis from the evidence of specific exposure history, clinical symptoms, and radiologic findings. The air exposure levels of the oil mist, endotoxin, total bacteria and fungus in the work environment was TWA(8-hr) 0.531 mg/m3, 6.33 EU/m3, 100 CFU/m3 and 75 CFU/m3, respectively. The concentrations of the endotoxin, total bacteria and fungus within the metal working fluid was 1.5x10(4) EU/mL, 4.6x10(5) CFU/mL and 1.8x10(5) CFU/mL, respectively. DISCUSSION: Although the patient did not receive a specific precipitating antibody test, the microbial concentration within the metal working fluid was higher than normal and similar to previous case reports, The oil mist level in the air exceeded the NIOSH REL and ACGIH NIC, and were similar or higher than previous cases. By excluding other causes of hypersensitivity pneumonitis, we concluded that the disease developed from exposure to microbial antigens in the metal working fluid.
Alveolitis, Extrinsic Allergic ; Bacteria ; Dyspnea ; Fungi ; Humans ; Hypersensitivity ; Korea ; Male ; National Institute for Occupational Safety and Health (U.S.)

INTRODUCTION: There have been no reports of hypersensitivity pneumonitis caused by metal working fluids in Korea, despite their existence in other countries. Here, we report the first such case, along with an assessment of work-relatedness through exposure assessment. CASE REPORT: A 64-year old male patient visited the hospital with dyspnea after metal pipe cutting for about a year. He was diagnosed with hypersensitivity pneumonitis from the evidence of specific exposure history, clinical symptoms, and radiologic findings. The air exposure levels of the oil mist, endotoxin, total bacteria and fungus in the work environment was TWA(8-hr) 0.531 mg/m3, 6.33 EU/m3, 100 CFU/m3 and 75 CFU/m3, respectively. The concentrations of the endotoxin, total bacteria and fungus within the metal working fluid was 1.5x10(4) EU/mL, 4.6x10(5) CFU/mL and 1.8x10(5) CFU/mL, respectively. DISCUSSION: Although the patient did not receive a specific precipitating antibody test, the microbial concentration within the metal working fluid was higher than normal and similar to previous case reports, The oil mist level in the air exceeded the NIOSH REL and ACGIH NIC, and were similar or higher than previous cases. By excluding other causes of hypersensitivity pneumonitis, we concluded that the disease developed from exposure to microbial antigens in the metal working fluid.
Alveolitis, Extrinsic Allergic ; Bacteria ; Dyspnea ; Fungi ; Humans ; Hypersensitivity ; Korea ; Male ; National Institute for Occupational Safety and Health (U.S.)

PURPOSE: The recently developed method of rapid immunohistochemistry (IHC) was applied to the intraoperative examination of sentinel lymph node (SLN) because as their routine frozen-section examination is liable to yield a false-negative results. This study is devoted to establish a reliable protocol for rapid IHC of SLN. METHODS: Between August 2004 and April 2005 a retrospective study was performed. SLNs from 50 breast cancer patients with clinically negative SLN were examined intraoperatively using hematoxylin-eosin (H&E) stain and immunostain for cytokeratin by rapid IHC assay. After examination of the frozen section, the SLNs were paraffin embedded and serially sectioned at 5 micrometer intervals. RESULTS: The median age and tumor size of the patients was 61.0 years and 1.4 cm (6% Tis, 70% T1, and 24% T2), respectively. The total number of dissected SLN was 112, with a mean of 2.2 (range, 1~4) SLNs per patient. Seven SLNs were found to be positive from metastasis in permanent pathological sections. Of these, 5 were stained by both intraoperative rapid IHC and H&E stain while one was not stained at all. The remaining SLN was initially stained with only the rapid IHC assay. The mean turn around time of the rapid IHC was less than 20 minutes, with sensitivity, true negative value, true positive values and accuracy of 85.7, 99.1, 100, and 99.1% respectively. CONCLUSIONS: The rapid IHC was a very sensitive and rapid technique for the intraoperative detection of metastatic involvement of SLNs, whitch may be helpful at increasing the accuracy of detecting the micro-metastasis of sentinel lymph nodes during an operation.
Breast Neoplasms* ; Breast* ; Frozen Sections ; Humans ; Immunohistochemistry* ; Keratins ; Lymph Nodes* ; Neoplasm Metastasis ; Neoplasm Micrometastasis ; Paraffin ; Retrospective Studies

OBJECTIVETo investigate brain activity patterns during acupuncture in stroke patients, and to compare the result with normal subjects using functional magnetic resonance imaging (fMRI).

METHODSA total of 11 stroke patients with motor weakness and 10 healthy subjects were studied. fMRI was performed during acupuncture on the left side at points Quchi (LI11) and Zusanli (ST36). Data were analyzed using statistical parametric maps of brain activation induced by acupuncture stimulation.

RESULTSThe results showed that stimulation of both LI11 and ST36 produced significantly different brain activation patterns between the two groups. The normal group showed a greater overall activation than the stroke group. In the normal group, parts of the frontal lobe, parietal lobe, sub-lobar, cerebellum and midbrain regions were activated by acupuncture at the left LI11. On the other hand, only the right side of the inferior parietal lobule region was activated in the stroke patients. When the left ST36 was stimulated in the normal group, both sides of the frontal lobe, parietal lobe, temporal lobe, and sub-lobar, and the left side of occipital lobe, and the right side of cerebellum and midbrain regions were activated. For the same stimulation in the stroke group, only both sides of the inferior parietal lobule and cerebellum regions were activated (P<0.05, cluster level). Deactivation pattern was not noted during any acupuncture stimulation in both groups.

CONCLUSIONBrain signal activations during the same acupuncture were different between the healthy and the stroke patients, and the effects showed a correlation of different acupuncture points.

Acupuncture Therapy ; Aged ; Brain Mapping ; Case-Control Studies ; Demography ; Female ; Health ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Stroke ; physiopathology

OBJECTIVESTo evaluate the effect of Uhwang Chungsim Won [UC, Niuhuang Qingxin Pill ()] on the blood oxygen level-dependent (BOLD) functional magnetic resonance imaging (fMRI) signal response to a motor stimulation task using a hand grasping paradigm.

METHODSTen healthy right-handed male volunteers were recruited. Their BOLD fMRI signal characteristics during hand grasping were determined before and 1 h after consuming UC.

RESULTSWith the administration of UC, BOLD responses to the right hand motor task showed a decrease in both hemispheres including the right cerebellar culmen, right and left medial frontal gyrus, right parahippocampal gyrus, right and left sublobar insula, right and left posterior cingulate, left precuneus, and right precentral gyrus. Decreased BOLD responses to the left hand motor task were shown in the right lingual gyrus, right cerebellar culmen, left superior temporal gyrus, right and left middle frontal gyrus, and right precentral gyrus after UC administration. Increased BOLD responses to the left hand motor task were shown in the left and right posterior cingulate, while there was no increased response to the right hand motor task after UC administration.

CONCLUSIONSUC administration significantly decreased fMRI BOLD signal responses to hand grasping in most of the brain areas including the primary motor cortex, but it is not clear whether these decreased responses were caused by a decrease in neuronal activities or by an increase in cerebral blood flow due to the cerebral vasodilatory effects of UC. Further studies using positron emission tomography or single photon emission computed tomography are needed to fully elucidate these findings.

Adult ; Brain Mapping ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Hand ; physiology ; Hand Strength ; physiology ; Humans ; Magnetic Resonance Imaging ; Male ; Motor Activity ; drug effects ; physiology ; Oxygen ; blood ; Signal Processing, Computer-Assisted ; Task Performance and Analysis

OBJECTIVE: To investigate the interaction between type II collagen (CII)-reactive T cell and fibroblast-like synoviocyte in rheumatoid arthritis (RA). METHODS: Peripheral blood T cells from RA patients were cultured with bovine CII and analyzed by flow cytometry. After co-culture with CII-reactive T cells and fibroblast-like synoviocytes (FLS), the expression of cytokines (IL-15 and TNF-alpha from FLS, IFN-gamma and IL-17 from CII-reactive T cells) were determined by ELISA and RT-PCR. RESULTS: CII-reactive T cells expressed CD69, one of the early activation markers, and produced significant amount of IFN-gamma, and proliferated. IL-15 and TNF-alpha expression from FLS were significantly elevated when co-culture with CII-reactive T cells and inhibited by physical interruption of cell-to-cell contact or anti-CD40 antibody. IFN-gamma and IL-17 expression from CII-reactive T cells were also significantly elevated when co-culture with FLS and inhibited by anti-IL-15 monoclonal antibody. CONCLUSIONS: CII-reactive T cells can activate FLS to secret proinflammatoy cytokines and interactions between these two cells drive further activation of each other. These data suggest that CII-reactive T cell may play a important role in pathogenesis of RA.
Arthritis, Rheumatoid* ; Coculture Techniques ; Collagen Type II ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Interleukin-15 ; Interleukin-17 ; T-Lymphocytes ; Tumor Necrosis Factor-alpha