Targeted therapies guided by molecular diagnostics have become a standard treatment of lung cancer. Epidermal growth factor receptor (EGFR) mutations and anaplastic lymphoma kinase (ALK) rearrangements are currently used as the best predictive biomarkers for EGFR tyrosine kinase inhibitors and ALK inhibitors, respectively. Besides EGFR and ALK, the list of druggable genetic alterations has been growing, including ROS1 rearrangements, RET rearrangements, and MET alterations. In this situation, pathologists should carefully manage clinical samples for molecular testing and should do their best to quickly and accurately identify patients who will benefit from precision therapeutics. Here, we grouped molecular biomarkers of lung cancers into three categories—mutations, gene rearrangements, and amplifications—and propose expanded guidelines on molecular testing of lung cancers.
Biomarkers ; Gene Rearrangement ; Humans ; Lung Neoplasms* ; Lung* ; Lymphoma ; Pathology, Molecular ; Phosphotransferases ; Precision Medicine ; Protein-Tyrosine Kinases ; Receptor, Epidermal Growth Factor

Wound healing is composed of a complex process that requires harmonies of various cell populations where fibroblasts play the main role. Oligomeric procyanidins (OPC) are main components of grape (Vitis vinifera) seed extracts, and recent studies showed OPC's effects on inflammation, cell migration, and proliferation. We investigated the effect of OPC on fibroblasts to regulate wound healing process. Human dermal fibroblast known as Hs27 cells were treated with various concentrations of OPC (0, 2.5, 5, 10, and 20 µg/µl). Cell cytotoxicity was evaluated by the Cell Counting Kit assay, and the expression levels of secreted procollagen were analyzed. Procollagen levels in OPC treated cells exposed to transforming growth factor beta 1 (TGF-β1) or ascorbic acid were evaluated using Western blot and immunocytochemistry. Relative mRNA expressions of procollagen, molecular chaperone such as HSP47, P4H were determined by real-time PCR in OPC treated cells. OPC showed no cytotoxicity on Hs27 cells at every concentration but inhibited procollagen secretion in a dose-dependent manner. The inhibitory effect also appeared under TGF-β1 induced collagen overproduction. Immunocytochemistry showed that higher levels of intracytoplasmic procollagen were accumulated in TGF-β1 treatment group, whereas ascorbic acid induced a release of accumulated procollagen under OPC treatment. The mRNA expressions of procollagen, molecular chaperone were not affected by OPC, but procollagen level was increased when exposed to TGF-β1. OPC inhibits procollagen secretion from fibroblasts with no effects on cell proliferations even under the environment of TGF-b1-induced collagen overproduction. OPC could regulate the diseases and symptoms of abnormal overabundant collagen production.
Ascorbic Acid ; Blotting, Western ; Cell Count ; Cell Movement ; Collagen ; Collagen Type I* ; Fibroblasts* ; Humans ; Immunohistochemistry ; Inflammation ; Molecular Chaperones ; Proanthocyanidins* ; Procollagen* ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; Transforming Growth Factor beta ; Vitis ; Wound Healing

BACKGROUND: This study evaluated the frequency and types of hand hygiene practices among healthcare workers directed by the WHO multimodal hand hygiene improvement strategy, and investigated the effect of hand hygiene practice on methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) isolation and MRSA acquisition rate and colonization pressure. METHODS: A quasi-experimental study was performed at a tertiary care university hospital with 850 beds from January to September 2012. We assessed the hospital hand hygiene program using the WHO hand hygiene self-assessment framework. The WHO multimodal strategy was used for healthcare workers with low indexes, and the subjects were reassessed. RESULTS: Hand hygiene compliance increased significantly from a pre-intervention rate of 58.7% to 72.6% post-intervention. MRSA and VRE isolation rates decreased from 1.69 per 1000 patient days to 1.41 and from 0.17 to 0.11, respectively. In intensive care units (ICUs), hand hygiene compliance rate rose to 77.9%, with a total score of 4.16 points out of 5 being awarded for the hand hygiene method, which was higher than that for the other care units. The pre-intervention MRSA acquisition rate in the ICU decreased from 7.47% to 4.30% post-intervention. This was associated with a decrease in the MRSA colonization pressure over the intervention period (26.2% to 16.9%). CONCLUSION: The utilization of the WHO multimodal strategy for improvement of hand hygiene increased the hand hygiene compliance rate and was effective in predicting a decreased rate of cross-infection, MRSA acquisition, and colonization pressure. We conclude that the implementation of such improvement strategies is crucial to maintaining hygiene standards and reducing infection within healthcare facilities.
Awards and Prizes ; Colon ; Compliance ; Delivery of Health Care ; Hand Hygiene* ; Humans ; Hygiene ; Intensive Care Units ; Methicillin-Resistant Staphylococcus aureus ; Self-Assessment ; Tertiary Healthcare

A case of peripheral primitive neuroectodermal tumor of the small bowel mesentery with osseous component is reported. A 23-year-old man was admitted to our hospital because of acute severe abdominal pain. Abdominal computed tomography revealed a large solid and cystic, oval shaped mass, measuring 11.0x6.0 cm in the pelvic cavity. Histologically the resected lesion consisted of sheets of undifferentiated small round cells forming Homer-Wright rosettes and perivascular pseudorosettes, and showed areas of osteoid and bone formation. Immunohistochemical studies revealed that tumor cells expressed positivity against CD99 (MIC2), CD57, neuron-specific enolase, and vimentin. Fluorescence in situ hybridization study revealed Ewing sarcoma breakpoint region 1 (EWSR1) gene rearrangement on chromosome 22q12. To the authors' knowledge this is the first documentation of a peripheral neuroectodermal tumor with osteoid and bone formation of the small bowel mesentery.
Abdominal Pain ; Fluorescence ; Gene Rearrangement ; In Situ Hybridization ; Intestine, Small ; Mesentery ; Metaplasia ; Neuroectodermal Tumors, Primitive ; Neuroectodermal Tumors, Primitive, Peripheral ; Osteogenesis ; Phosphopyruvate Hydratase ; Sarcoma, Ewing ; Vimentin

BACKGROUND: Recently, BRAF inhibitors showed dramatic treatment outcomes in BRAF V600 mutant melanoma. Therefore, the accuracy of BRAF mutation test is critical. METHODS: BRAF mutations were tested by dual-priming oligonucleotide-polymerase chain reaction (DPO-PCR), direct sequencing and subsequently retested with a real-time PCR assay, cobas 4800 V600 mutation test. In total, 64 tumors including 34 malignant melanomas and 16 papillary thyroid carcinomas were analyzed. DNA was extracted from formalin-fixed paraffin embedded tissue samples and the results of cobas test were directly compared with those of DPO-PCR and direct sequencing. RESULTS: BRAF mutations were found in 23 of 64 (35.9%) tumors. There was 9.4% discordance among 3 methods. Out of 6 discordant cases, 4 cases were melanomas; 3 cases were BRAF V600E detected only by cobas test, but were not detected by DPO-PCR and direct sequencing. One melanoma patient with BRAF mutation detected only by cobas test has been on vemurafenib treatment for 6 months and showed a dramatic response to vemurafenib. DPO-PCR failed to detect V600K mutation in one case identified by both direct sequencing and cobas test. CONCLUSIONS: In direct comparison of the currently available DPO-PCR, direct sequencing and real-time cobas test for BRAF mutation, real-time PCR assay is the most sensitive method.
DNA ; Humans ; Indoles ; Melanoma ; Paraffin ; Real-Time Polymerase Chain Reaction ; Sulfonamides ; Thyroid Neoplasms

BACKGROUND: An epidemiologic study was performed after the outbreak of carbapenem-resistant Acinetobacter baumannii (CRAB) in the medical intensive care unit (MICU) from December 2006 to May 2007. METHODS: A retrospective case-control study was performed using the medical records of the patients. The case and control patients were compared for age, gender, total length of stay in MICU, prior carbapenem use, Acute Physiology and Chronic Health Evaluation II (APACH II) score, presence of central line, effect of mechanical ventilation, and sputum suction. Environmental and hand-washing studies were performed during the outbreak. RESULTS: Ten CRAB-affected patients and 29 controls were enrolled in this study. Univariate analysis showed that the age, total length of stay in MICU, presence of central line, and prior carbapenem use were associated with the CRAB outbreak. However, multivariate analysis showed that only prior carbapenem use was associated with the CRAB outbreak (odd ratio: 8.67, P=0.01). The outbreak disappeared after implementing a combined infection control strategy, including the sequential disinfection of MICU and strict compliance with cross-transmission prevention protocols. CONCLUSION: The use of carbapenem was associated with an increased risk of CRAB infection. This study suggests that the MICU contamination and infection transmission by health-care workers played a major role in the CRAB outbreak. Novel strategies such as restricted use of broad-spectrum antibiotics, strict hand hygiene, strict isolation of the patients, and MICU disinfection may be required to prevent the CRAB outbreak.
Acinetobacter ; Acinetobacter baumannii ; Anti-Bacterial Agents ; APACHE ; Case-Control Studies ; Compliance ; Disease Transmission, Infectious ; Disinfection ; Epidemiologic Studies ; Hand Hygiene ; Humans ; Infection Control ; Critical Care ; Intensive Care Units ; Length of Stay ; Medical Records ; Multivariate Analysis ; Respiration, Artificial ; Retrospective Studies ; Risk Factors ; Sputum ; Suction

The significant advance in the development of molecular-targeting drugs has made an evaluation of Her-2, EGFR, and cyclin D1 an important clinical issue in breast cancer patients. This study compared the Her-2, EGFR, and cyclin D1 status of primary tumors as well as their matching lymph node metastases using immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH) in 73 breast cancer patients. Her-2, EGFR, and cyclin D1 protein showed a concordance between the primary lesion and the metastatic regional lymph nodes in 82%, 90%, and 63%, respectively. CISH also revealed 92%, 93%, and 85% concordance in the gene amplification status of Her-2, EGFR, and cyclin D1, showing a reasonable agreement between primary tumors and metastatic regional lymph nodes. Although a statistically significant agreement was found in Her-2 expression, a relatively high discordance rate (18%) raises a little concern. Our findings suggest that the Her-2 status can be reliably assessed on primary tumor but a possible difference can be found in Her-2, EGFR, and cyclin D1 status between the primary and the metastatic sites and this possibility should be concerned in patients considering molecular targeted therapy or patients with progress of disease.
Adult ; Aged ; Breast Neoplasms/genetics/*metabolism/pathology ; Chromogenic Compounds ; Cyclin D1/*analysis/genetics ; Female ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Lymph Nodes/*metabolism/pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Recurrence, Local/genetics/metabolism ; Receptor, Epidermal Growth Factor/*analysis/genetics ; Receptor, erbB-2/*analysis/genetics ; Survival Analysis

OBJECTIVE: CD27 is a member of the tumor necrosis factor receptor (TNFR) superfamily and is expressed on T, B, and NK cells. The signaling via CD27 plays pivotal roles in T-T and T-B interaction. CD27 is a useful marker in assessing the number of circulation B cells and B cell subsets because it permits one step identification of the major B cell compartments, CD27- naive and CD27+ memory B cells as well as CD27high plasma cells. We have analyzed the mechanisms underlying the regulation of CD27 expression. METHODS: Isolation B cells and Raji cells were cultured with PMA. The levels of cell surface CD27 and CD 27 mRNA were analyzed by FACs staining and RT-PCR. Raji cells were cultured with phorbol 12-myristate 13-acetate (PMA), with or without pretreated shedding inhibitor BB3103 and TAPI-1. sCD27 was measured in culture supernatant by ELISA. Cell lysates were analyzed for PKC isotype activation by Western blot. We used PKC inhibitor Ly379196 and rottlen. RESULTS: Membrane expression of CD27 was down-regulated after PMA stimulation without cytotoxic effect in B cells. Furthermore, PMA treatment could directly reduce CD27 mRNA without intermediate protein synthesis in B cells. In contrast, PMA treatment induced soluble form of CD27 (sCD27), which was shed from the cell surface and was found in PMA treatment B cell culture supernatant. PMA-induced sCD27 proteins were decreased with shedding inhibitor BB3103 and TAPI-1. PMA-induced down regulation of CD27 expressions were quenched with protein kinase C (PKC) inhibitor Staurosporin, PKC-beta inhibitor rottlerin and PKC-delta inhibitor Ly379196. CONCLUSION: These data suggest that PMA-induced activation of PKC plays a crucial role in down-regulation of the expression of the CD27 and up-regulation of the shedding of the CD27 in human B cells.
B-Lymphocyte Subsets ; B-Lymphocytes* ; Blotting, Western ; Cell Culture Techniques ; Down-Regulation* ; Enzyme-Linked Immunosorbent Assay ; Humans ; Killer Cells, Natural ; Membranes* ; Memory ; Plasma Cells ; Protein Kinase C ; Protein Kinase C-delta* ; Protein Kinases* ; Receptors, Tumor Necrosis Factor ; RNA, Messenger ; Up-Regulation

BACKGROUND: CD24 was originally described as a B cell-specific marker, however its aberrant expression in various solid tumors has recently been reported. Our objective was to determine the pattern and extent of the CD24 expression in colorectal cancer and its related lesions, and to clarify its correlation with clinico-pathological parameters and especially those associated with patients' prognoses. METHODS: A total of 307 colorectal cancers and the related lesions (150 carcinomas, 30 high-grade adenomas, 49 low-grade adenomas, 41 hyperplastic polyps, and 37 normal colorectal epithelia) were immunohistochemically analyzed by treating CD24 monoclonal antibody onto tissue embedded paraffin blocks. RESULTS: CD24 expression was very rarely observed in the normal epithelia, hyperplastic polyps, and low-grade adenomas; however, in high-grade adenomas, the CD24 expression was shown to be mildly increased in the cytoplasm (13.3%). In carcinomas, the CD24 expression was increased substantially in both the membrane (38.0%) and the cytoplasm (44.7%). The expression of CD24 in the membrane was positively correlated with tumor size (p<0.01). The CD24 expression in the cytoplasm was positively correlated with several unfavorable parameters, including a larger tumor size (p<0.01), a higher tumor grade (p<0.01), a higher rate of tumor invasion (p<0.05), and a higher pTNM stage (p<0.05). CONCLUSION: High levels of CD24 expression in the membrane and cytoplasm were characteristic in colorectal cancer, and the cytoplasmic CD24 expression was correlated with several unfavorable clinical parameters.
Adenoma ; Colorectal Neoplasms* ; Cytoplasm ; Immunohistochemistry ; Membranes ; Paraffin ; Polyps ; Prognosis

Background: The shelf life policies for central supply department (CSD) sterilized items and other devices should be determined by the healthcare facility's infection control program. We investigated effect of the sterility integrity of the CSD sterilized packs by wrapping-materials, storage period and environment to modify and extend current shelf-life. Methods: The first phase study was from May to October in 2000 and the second phase study was planned to extend further the shelf-life of the sterile packs from April 2001 to June 2002. Six hundred and fourty packs containing small gauze with four wrapping materials(100 times and 50 times washed two-ply reusable cotton, disposable craft paper, and disposable new pouch bag) and the 104 returned set after their shelf-life were stored on the top or middle of shelves or closed cabinets and storage durations from 1 to 20 weeks in the first phase study. The test packs were collected weekly and cultured in the laboratory. Five hundred seventy-six test packs were prepared with three wrapping materials (except 50 times washed cotton and returned set) and stored in the same location as the First phase study and collected and cultured monthly after three months storage (from July 2001) for one year in the second phase study. The temperature and relative humidity was monitored whenever the test pack was collected. Results: The gauze in the test packs were not contaminated until 154 days in the first study phase and until 423 days in the second phase study. The temperature and relative humidity of storage locations were 25.9degrees C and 55.2% in the first phase study and 26.0degrees C and 45.9% in the second phase study, respectively. Conclusions: There was no difference in the sterility integrity of the test packs with different wrapping materials. storage locations and environments. and storage durations. It was possible to extend shelf-life from two weeks to three and six months in the study hospital.
Delivery of Health Care ; Humidity ; Infection Control ; Infertility