<原著>軟骨細胞の分化における HSP70 の発現

To know the role of HSP70 in chondrocytes, HSP70 expressions under heat stress or in a nonstress condition were examined by using electrophoresis, immunohistochemistry, and in situ hybridization. As a result, HSP70 was observed in proliferating chondrocytes in the micro-mass cultures without heat stress. In this culture, chondrocytes maintained the terminal differentiation potency. On the other hand, HSP70 did not appear in the chondrocytes in the logarithmic growth phase of the monolayer culture. In growth plates in vivo, HSP70 expressions in the chondrocytes located in the resting and hypertrophic zones were observed with immunohistochemistry. Appearance of HSP70 mRNA was also confirmed by in situ hybridization in the proliferating zone of growth plate. HSP70 can be expressed not only in chondrocytes under heat stress but also in the cells without stress, and the expression would be related to the terminal differentiation of chondrocytes. HSP70 is surmised to promote hypertrophy and calcification by stopping protein synthesis in chondrocytes which possess terminal differentiation potency.軟骨細胞との関連が注目されている熱ショック蛋白質の発現を, 遠沈管培養軟骨細胞と骨端軟骨板を用いて, in vitro および in vivo で検索した. 温熱ストレス下のラﾂト軟骨細胞では, 少なくとも3種類の熱ショック蛋白質の発現がみとめられ, そのうち HSP70 の誘導量が最大であった. 非ストレス下では, 遠沈管培養軟骨細胞のうち肥大軟骨細胞様細胞への分化能を有すると考えられる細胞に HSP70 の発現が認められた骨端軟骨板においては, 肥大軟骨細胞で HSP70 が誘導されていた. また, HSP70 mRNA は肥大層に隣接する増殖層および静止層の軟骨細胞で観察された. このことは, 肥大軟骨細胞への分化能を有する軟骨細胞では, すでに HSP70 mRNA が誘導されていることを示している. これらの結果から, HSP70 の発現が軟骨細胞の分化と関係していることが推察された

Changes in Attitudes of Japanese Doctors toward Complementary and Alternative Medicine—Comparison of Surveys in 1999 and 2005 in Kyoto

We surveyed the attitudes of Japanese medical doctors toward complementary and alternative medicine (CAM) in 1999. It is supposed that the situation concerning CAM has been changing recently. The aim of the present study is to survey the attitude of doctors toward CAM again, and to examine changes in attitude over the last 6 years. The attitudes of medical doctors belonging to the Kyoto Medical Association toward CAM were surveyed by a structured, self-administered questionnaire in 1999 and 2005. The results showed that the doctors familiar with the term “CAM”, practicing CAM therapies, and attending meetings or training courses related with CAM, increased significantly from 1999 to 2005. The doctors who possessed knowledge of CAM also increased significantly from 1999 to 2005. Almost all doctors believed in the effectiveness of Kampo (Japanese traditional herbal medicine) and acupuncture. The number of doctors who believed in the effectiveness of aromatherapy and ayurveda increased significantly in 2005, compared with 1999. In the near future, 58% of doctors desired to practice CAM therapies. In conclusion, the numbers of doctors who practice CAM therapies, possess CAM knowledge and desire to practice such therapies have increased over the last 6 years in Japan

Successful transfer of ADA gene in vitro into human peripheral blood CD34+ cells by transfecting EBV-based episomal vectors

AbstractWe report a novel non-viral system for transfecting human immature hematopoietic cells in vitro. Epstein-Barr virus (EBV)-based episomal vectors carrying human adenosine deaminase (ADA) gene cDNA were transfected by electroporation into human peripheral blood (PB) CD34+ cells. The transgene-specific mRNA were detected from 37 to 100% of CFU-c (colony forming unit in culture) colonies derived from the transfected cells. A two-fold increase in enzyme activity was also found. These results indicate the successful transfer and expression of genes in human immature hematopoietic cells using the EBV-based episomal vector system

Analysis of heat shock proteins and cytokines expressed during early stages of osteoarthritis in a mouse model

SummaryObjective:Osteoarthritis (OA) is a debilitating disease of the joints. The joints of affected individuals are characterized by a progressive degeneration of articular cartilage leading to inflammation and pain. The expression of heat shock proteins (HSPs) is a ubiquitous self-protective mechanism of all cells under stress, furthermore, the synovium of osteoarthritic individuals contains high levels of cytokines. This study seeks to establish the role of HSPs and cytokines in OA.Methods:We have investigated the presence of HSPs and cytokines in articular cartilage during early stages of OA in a mouse that is known to develop spontaneous OA lesions (C57 black mouse). The articular cartilage from closely related mice (C57BL/6) was used as control. Messenger RNAs (mRNAs) for HSPs (HSP32, HSP47, HSP60, HSP70, HSP84 and HSP86) and cytokines [interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ)] were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results:The mRNA levels of HSP47, HSP70, HSP86, IL-6, and IFN-γ were up-regulated in the cartilage of C57 black mice, whereas, the level of expression of HSP32, HSP60, HSP84 and IL-1β remained unchanged. Furthermore, the expression of IL-1β, IL-6, TNF-α and IFN-γ mRNA was associated with expression of HSP60, HSP47, HSP70 and HSP70/HSP86 mRNA, respectively.Conclusions:The findings in this study suggest that chondrocytes are conditioned under non-physiological stress during early stages of OA, In addition, among HSPs, HSP70 was associated with two different highly expressed cytokines in C57 black mice, indicating the possible role of HSP70 as a characteristic indicator of early stage of OA

Immunological and Psychological Benefits of Aromatherapy Massage

This preliminary investigation compares peripheral blood cell counts including red blood cells (RBCs), white blood cells (WBCs), neutrophils, peripheral blood lymphocytes (PBLs), CD4(+), CD8(+) and CD16(+) lymphocytes, CD4(+)/CD8(+) ratio, hematocrit, humoral parameters including serum interferon-γ and interleukin-6, salivary secretory immunoglobulin A (IgA). Psychological measures including the State–Trait Anxiety Inventory (STAI) questionnaire and the Self-rating Depression Scale (SDS) between recipients (n = 11) of carrier oil massage and aromatherapy massage, which includes sweet almond oil, lavender oil, cypress oil and sweet marjoram oil. Though both STAI and SDS showed a significant reduction (P < 0.01) after treatment with aromatherapy and carrier massage, no difference between the aromatherapy and control massage was observed for STAI and SDS. Aromatherapy, in contrast to control massage, did not significantly reduce RBC count or hematocrit. However, aromatherapy massage showed a significant (P > 0.05) increase in PBLs, possibly due to an increase in CD8(+) and CD16(+) lymphocytes, which had significantly increased post-treatment (P < 0.01). Consequently, the CD4(+)/CD8(+) ratio decreased significantly (P < 0.01). The paucity of such differences after carrier oil massage suggests that aromatherapy massage could be beneficial in disease states that require augmentation of CD8(+) lymphocytes. While this study identifies the immunological benefits of aromatherapy massage, there is a need to validate the findings prospectively in a larger cohort of patients

Anxiolytic Effect of Aromatherapy Massage in Patients with Breast Cancer

We examined how aromatherapy massage influenced psychologic and immunologic parameters in 12 breast cancer patients in an open semi-comparative trial. We compared the results 1 month before aromatherapy massage as a waiting control period with those during aromatherapy massage treatment and 1 month after the completion of aromatherapy sessions. The patients received a 30 min aromatherapy massage twice a week for 4 weeks (eight times in total). The results showed that anxiety was reduced in one 30 min aromatherapy massage in State-Trait Anxiety Inventory (STAI) test and also reduced in eight sequential aromatherapy massage sessions in the Hospital Anxiety and Depression Scale (HADS) test. Our results further suggested that aromatherapy massage ameliorated the immunologic state. Further investigations are required to confirm the anxiolytic effect of aromatherapy in breast cancer patients

Cyclic AMP Responsive Element Binding Proteins Are Involved in ‘Emergency’ Granulopoiesis through the Upregulation of CCAAT/Enhancer Binding Protein β

In contrast to the definitive role of the transcription factor, CCAAT/Enhancer binding protein α (C/EBPα), in steady-state granulopoiesis, previous findings have suggested that granulopoiesis during emergency situations, such as infection, is dependent on C/EBPβ. In this study, a novel lentivirus-based reporter system was developed to elucidate the molecular switch required for C/EBPβ-dependency. The results demonstrated that two cyclic AMP responsive elements (CREs) in the proximal promoter region of C/EBPβ were involved in the positive regulation of C/EBPβ transcription during granulocyte-macrophage colony-stimulating factor (GM-CSF)–induced differentiation of bone marrow cells. In addition, the transcripts of CRE binding (CREB) family proteins were readily detected in hematopoietic stem/progenitor cells. CREB was upregulated, phosphorylated and bound to the CREs in response to GM-CSF stimulation. Retroviral transduction of a dominant negative CREB mutant reduced C/EBPβ mRNA levels and significantly impaired the proliferation/differentiation of granulocyte precursors, while a constitutively active form of CREB facilitated C/EBPβ transcription. These data suggest that CREB proteins are involved in the regulation of granulopoiesis via C/EBPβ upregulation

舌転移をきたした腎細胞癌の1例

We report a case of renal cell carcinoma (RCC) metastatic to the tongue in a 58-year-old female. The patient had undergone radical nephrectomy for renal cell carcinoma 5 years previously, and experienced multiple metastatic disease in lung, bone, and contralateral kidney, before she noticed yellowish tumor on the left border of the tongue. Microscopic appearances of the biopsied lingual tumor were almost identical to those of the primary kidney tumor, thus the diagnosis of lingual metastasis from renal cell carcinoma was established. This case represents the tenth case of RCC metastatic to the tongue