変形性膝関節炎患者膝関節液中の細胞外マトリックス成分と可溶性VCAM-1

It is generally accepted that the number of patients with osteoarthritis (OA) of the knee must increase in the future because of the increase of the aged. The pathogenic mechanisms of OA haven't been elucidated yet, but it is supposed that mediate pannus may cause the factors for irreversible cartilage damage in the inflammatory process. Therefore it is very important to find and predict OA at the early stage. Now there were many indices to determine joint damage with OA, for example, cytokines including IL, TNF, TGF, matrix metalloproteinase (MMPs), tissue inhibitor of metalloproteinase (TIMPs), proteoglycan (PG), hyaluronate (HA), super oxide, adhesion molecules including, ICAM-1, VCAM-1 and PECAM. In this study, we measured that the soluble VCAM-1 (sVCAM-1), soluble collagen (sCOLL), heparansulfate proteoglycan (HSPG), uronic acis and CRP in synovial fluids (SF) with OA. A positive correlation between the level of sVCAM-1 and sCOLL (ρ = 0.652) was noted. However there was no factor showing a correlation with CRP. It was suggested that the levels of sVCAM-1 and sCOLL in the synovia fluid were active inflammation indices of knee damage, as the different aspects from CRP. The level of HSPG also showed a unique trend because HSPG on the cell surface was quickly digested endocytosis, so that it was not suitable for the inflammatory index in synovial fluid with OA.変形性膝関節炎(OA)患者は,高齢化社会の到来とともに今後ますます増加するものと考えられる｡OAの発症メカニズムについては,関節軟骨の破壊･修復やパンヌス形成に関わる多くの関連因子(サイトカイン,マトリックス分解酵素,分解酵素抑制因子,プロテオグリカン,活性酸素,細胞接着因子など)について,多面的に研究が進んでいる｡しかし,膝関節の炎症初期の病態のメカニズムについては,なお詳細に解明されておらず,関節炎初期の炎症指標の検索が今後さらに必要と考えられる｡本研究で私たちは, OA患者膝関節液中の細胞外マトリックス構成成分であるコラーゲン,へパラン硫酸プロテオグリカン,ウロン酸および炎症の最も初期に血管内皮細胞に出現するVCAM-1等の可溶化成分濃度を測定した｡患者膝関節液(SF)中の可溶性コラーゲン量と可溶性VCAM-1(sVCAM-1)量とは有意な正の相関関係が(ρ=0.652)観察された｡しかし,ヘパラン硫酸プロテオグリカン(HSPG)と可溶性コラーゲン(sCOLL)値間には有意な相関関係は確認されなかった｡また,一般に炎症の指標として用いられるCRP値とも比戟検討したが,CRP値と他の測定項目値とは一定の相関関係は存在しなかった。以上の結果より,OA患者のSF中の炎症マーカ-として,急性炎症時には従来の炎症指標蛋白質であるCRP値が有力であるというSipe JDの報告から推測するとsCOLLとsVCAM-1はCRP値と一定の相関を示さないことより,CRPの動態とは異なった炎症指標としての有用性が考えられる｡しかし,HSPGはエンドサイトシスにより速やかに代謝されるためSF中への解離が少なく,炎症指標としての可能性は低いと考えられる

The Units of IgM, IgA and IgG Rheumatoid Factors by Enzyme Linked Immunosorbent Assay in Human Sera

The measurement of Rheumatoid Factor (RF) has been a valuable tool in diagnosing Rheumatoid Arthritis (RA) for many years. Since Waaler's discovery of the RF, many studies have been carried out. In the recent years studies have been shown that there is no such correlation between the level of the RF titers and disease activity. In this report we have measured the IgM-RF, IgA-RF and IgG-RF by an Enzyme linked immunosorbent assay (ELISA) for 144 normal human sera and 50 patients sera. At the same time we have measured the titers of gelatin particle agglutination (RAPA) and the Latex agglutination (RA-test). The value of IgG-RF have been most correlated to the titers of RAPA (r=0.9037). Using this ELISA technic to detect IgM-, IgA- and IgG-RF, have been aviable more clearly the activity and the stage of Rheumatoid disease

リウマチ因子陽性ヒト血清中の免疫グロブリンGクラスリウマチ因子と免疫複合体ならびにCRPとの相関性

We measured the concentration of three immunoglobulin classes of rheumatoid factors (Ig-RFs), C3d binding IgG immune complex (C3d-IC), C1q binding IgG immune complex (C1q-IC) and C-reactive protein (CRP) in 74 samples of human sera with high levels of RF (24.0~2350.0IU/ml). In sera with high levels of C3d-IC (>15.0μg/ml), there was a positive correlation between the levels of CRP and the IgG-RF, but there was no correlation between the levels of CRP and the immune complexes (C3d-IC and C1q-IC). And then, there was a positive correlation between the levels of CRP and IgG-RF or C3d-IC and IgG-RF when the levels of C1q-IC in patients sera were higher than 80.0μg/ml. However, there was no correlation between the levels of CRP and C1q-IC in these patients sera containing high levels of both C3d-IC and C1q-IC. These results indicated that the determination of C3d-IC, C1q-IC, IgG-RF and CRP in human sera containing RF denote different implications as inflammatory indexes on progression of rheumatoid arthritis and other autoimmune diseases.一般に慢性関節リウマチの診断ならびに活動性の一指標として、日常臨床検査では患者血清中のリウマチ因子の測定が実施されている。本論文ではRF検査依頼が有り、高RF値（24.0～2350.0IU/ml）を示した患者血清74検体について、免疫グロブリンクラス別RF値を測定するとともに自己IgGと補体との免疫複合体（IC）であるC1q-IgG・IC（C1q-IC）、C3d-IgG・IC（C3d-IC）ならびにCRP値を測定し、それらの測定値間の相関性について統計学的検討を行った。C3d-IC値が15.0μg/ml以上の高値を示す患者血清では、CRP値とIgGクラスのRF（IgG-RF）値間については正の相関結果がが得られた。また、C1q-IC値が80.0μg/ml以上の場合にもCRP値とIgG-RF値間およびC3d-IC値とIgG-RF値間では正の相関結果が得られた。しかし、C3d-ICならびにC1q-ICともに高値例の患者血清中のCRP値とC1q-IC値間には有意な相関は観察されなかった。以上の結果より、慢性関節リウマチの活動度を判定する上で、従来から炎症マーカーとしてCRP値が利用されているが、血清中に免疫複合体が高レベルに検出される患者については、炎症の指標としてCRP以外にもC1q-ICやC3d-ICおよびIgG-RF等を加えた総合的な判断が必要であることが示唆される

長期ホルマリン固定により失活したProliferating Cell Nuclear Antigen (PCNA) の免疫反応性回復条件の基礎的検討 ―マイクロウェーブ、オートクレーブの影響について―

Using paraffin-embedded tissue sections of liver cancer obtained from autopsy which had been preserved in 10% buffered formalin solution for 6 months while PCNA immunoreactivity was lost, we examined the effects of heat processing by either microwave(MW) and autoclave(AC) in the presence of various processing solution. It appeared that AC processing took shorter time period than MW irradiation to restore equal immunoreactivity. With regard to immunoreactivity retrieval by MW irradiation,however, variation of the degree of retrieval depending on processing time was smaller than in AC, and so the stable consequences were obtained. Although AC processed tissues tended to be stained deep, prolonged processing time presented strong background staining and blurred nuclear margins which made it difficult to estimate the positive cell count. As for the effects of processing solution, there was little difference in retrieval of PCNA among 0.01 M citrate buffer (pH 6.0), saturated solution of lead thiocyanate and distilled water, but the least background staining was observed with distilled water. These observations suggest that MW irradiation of which effect of retrieval is less dependent of processing time and with the least background stainability, is superior to AC processing for PCNA immunoreactivity retrieval on formalin-fixed tissues.10％緩衝ホルマリンに6ヶ月間浸漬していた剖検材料（肝臓癌）のパラフィン包埋後の組織切片を用いて、ホルマリンの固定作用により失活したPCNAの免疫反応性の回復にマイクロウェーブ（MW）及びオートクレーブ（AC）による熱処理とその時用いる処理溶液が、どのような影響を与えるかについて検討した。その結果、同等の免疫反応性を回復するのには、AC処理の方がMW照射より短時間でよいことが解った。しかし、MW照射による免疫反応性回復では、処理時間による影響がAC処理に比べて少なく、安定した結果が得られた。また、PCNAの染色所見については、AC処理の方が濃く染まる傾向が見られた。しかし、処理時間が長くなるとバックグラウンドの染色性が高くなる、核の周囲ににじみ現象が見られる等の所見があり判定に困難をきたした。一方、処理溶液についてはクエン酸（0.01M pH6.0）、チオシアン酸鉛飽和溶液、蒸留水について検討を行った結果、PCNAの回復には差は認められなかったが、バックグラウンドの染色については蒸留水が最も少なかった。以上の結果により、ホルマリン固定により失活したPCNAの免疫反応性を回復するには、MW照射の方がAC処理より処理時間に関係なく安定した染色性が得られ、なおかつバックグラウンドの染色性が少ない等の点で優れていることが示唆された

ヒト血清リポタンパク質とシクロデキストリンのアガロースゲル内における相互作用

Cyclodextrins (CDs) are cyclic oligosaccharides with low molecular weight. They have been known to bind to some serum lipoproteins and to form complexes. To elucidate whether each serum lipoprotein subclass could be separated by electrophoresis using CDs, we performed preliminary experiment, in which lipoprotein-CDs interaction was examined on electrophoresis with agarose gel. When the supporting agarose gel containing both α-CD and β-CD was prepared and was applied to isoelectric focusing for fractionating serum lipoproteins, apoB lipoproteins were found to be clearly separated into several fractions on this electrophoresis. This finding suggested that apoB lipoprotein may be detected as isolated form by arranging amounts of added CDs.シクロデキストリンは低分子量の環状オリゴ糖で, リポタンパク質と複合体を形成する｡シクロデキストリンを使用した電気泳動による血清リポタンパク質サブクラスの分離の可能性を明らかにするために, リポタンパク質とシクロデキストリンとの泳動用ゲル内での相互作用について検討をおこなった｡その結果,α-CDとβ-CDの2種類のCDをアガロースゲルに添加し,このゲルを泳動用支持体として等電点電気泳動をおこなったところ,apoB含有リポタンパク質が数分画分離された｡したがって,ゲル内へのシクロデキストリンの適正な添加条件が設定できるならば,apoB含有リポタンパク質サブクラスの分離が可能となることが示唆された

IgG Heavy-Chain Subclass Typing of Human Serum M-Protein -Optimal condition for the separation and the detection with high resolution of agarose gel isoelectrofocusing-

IgG Heavy-Chain subclass typing of human serum M-protein were isoelectrofocussed (IEF) in agarose gel, and then the bands of IgG were detected with peroxidase-conjugated anti-human IgG1-4 monoclonal antibodies on the same isoelectrofocussed agarose gel plate. This IEF enzyme immunodetection patterns were composed of four to eight discrete bands (The range of pI was 6.0 to 9.0). These bands were dependent on immunofixationbands. It was very specific and clear to detect the subclass of IgG Type M-protein in human serum using this method

Views concerning nursing and nursing course of senior high school guidance teachers

1993年に岡山県で行った先の研究で,高校生の看護課程の選択にあたって,進路指導を行っている教師の役割が大きい事を報告した｡今回は3年経った1996年に,同じ郵送調査を岡山県および周辺10県の普通科高校617校より無作為に選んだ127校において実施した｡64校より得られた回答を要約すると以下の通りであった｡1.教師が挙げた看護婦不足の理由はまず｢勤務時間が厳しい(85.9%)｣,｢仕事の内容が厳しい(53.1%)｣といった仕事のきつさで,次いで｢仕事の割には社会的評価が低い(32.8%)｣であった｡2.教師が学生に肯定的イメージとして看護課程を勧める理由は,｢まず専門職であること(73.4%)｣,次いで｢人に貢献できる喜び(53.1%)｣であった｡これらの結果は前回の結果と比較して統計的な有意差は認めなかった｡3.近年進められている3年制看護課程(準学士課程)から4年制課程(学士課程)への移行に関して,教師はこれらは学生の関心を呼ぶとし(79.8%),看護課程へ進学する者は増加するであろうとしている(57.8%)｡またこれまでよりこの課程への進学を教師は勧め易くなるとしている(56.3%)。これらの率は前回の結果と比較すると統計的に有意に高かった。A previous study in Okayama prefecture in 1993 indicated the importance of high school teachers' influential rule on student's choice of nursing course. The same questionnaire survey was sent out by mail in 1996 to guidance teachers of 127 general high schools randomly selected from 617 schools in 10 prefectures, in and around Okayama. Data obtained from 64 teachers were analyzed and the results were as follows; 1. They suggested that shortage of nursing manpower was due to the harsh working conditions such as long and irregular hours (85.9%) and hard work (53.1 %), and also the low social esteem of nurse (32.8%). 2. The aspect of nursing noted by teachers were job specialty (73.4%) and devotion to people (53.l%). There were no statistical difference between these results and those of the previous study. 3. The teachers answered that the new four-year nuersing course (baccalaureate) currently being promoted from over the the present three-year course (associate degree) would be an incentive to students (79.8%) and might increase the number of students choosing to study nursing (57.8%). And they also noted that they would more readily recommend this course to students than before. These rates were significantly increased compared to the results of the previous report

Comparative studies of proliferating cell nuclear antigen and bromodeoxyuridine staining pattern in rat regenerating liver cells

種々の条件下（10％緩衝ホルマリン・6時間、1％パラホルムアルデヒド・6時間、100％メタノール・4時間、70％エタノール・6時間、100％アセトン・2時間、カルノア液・4時間）で固定したラット再生肝組織に免疫組織染色を施し、Proliferating cell nuclear antigen （PCNA）の核内染色パターンについての検討を行った。その結果、100％メタノール及び70％エタノールで固定した場合には、明瞭な顆粒状の染色像が得られた。10％緩衝ホルマリン及び1％パラホルムアルデヒドの場合には、明瞭な顆粒状並びに弱いびまん性の染色像が得られた。S期に対する代表的なマーカーとして知られるBromodeoxyuridine （BrdU）の染色像との比較のもと、再生肝組織中のPCNA陽性細胞の数とBrdU陽性細胞の数を再生ピーク時の術後1日目で検討したところ、100％メタノール及び70％エタノールで固定した際の陽性細胞数が、最もBrdUの所見と一致した。しかし、10％緩衝ホルマリン、1％パラホルムアルデヒド、100％アセトン、カルノア液で固定した場合には、BrdUの所見と著しい解離が見られた。これらの知見は、PCNAを指標に細胞増殖を評価する上で、100％メタノール、70％エタノールでの固定が有用であることを示唆する。Immunohistochemical staining pattern of proliferating cell nuclear antigen (PCNA) was examined in rat regenarating liver specimens fixed under the various conditions (10% buffered formalin for 6 hr, 1% paraformaldehyde for 6 hr, 100% methanol for 4 hr, 70% ethanol for 6 hr, 100% acetone for 2 hr and Carnoy solution for 4 hr). Most distinct granular staining pattern of PCNA was observed in fixation with 100% methanol or 70% ethanol. Specimens fixed with 10% buffered formalin or 1% paraformaldehyde showed both distinct granular and slight diffuse pattern of PCNA staining. Specimens fixed with 100% acetone showed only slight diffuse staining. Specimens fixed with Carnoy solution showed little staining. Number of PCNA positive cells in the specimens fixed with 100% methanol or 70% ethanol on 24 hours after hepatectomy coincided closely with those of BrdU positive cells, which is known to be a representative marker of cell proliferation. In contrast, dissociative result was obtained in specimens fixed with 10% buffered formalin, 1% paraformaldehyde, 100% acetone or Carnoy solution when numbers of PCNA positive cells were compared with those of BrdU. These findings suggest that tissue specimens fixed with 100% methanol or 70% ethanol is useful for evaluating the extent of cell proliferation

Senior high school students' recognition and interest in allied medical education of college

1996年7～9月にかけて,岡山県周辺の10県下高校普通科3年在学中の学生に志望進路,志望分野とともに大学,短期大学における医療技術教育への認識と関心についてアンケート調査を行った｡サンプリングによって依頼した127校のうち回答が得られたのは52校(回収率40.9%)で,得られた回答数は合計1,998人(男子891人,女子1,077人,不明30人)であった｡これらの分析から,これらの地域では進学志望者の率は高く,特に男子に著名で,男女ともに理系への志望が多いことを認めた｡医療技術系大学の認識は看護学科を除いて低く,関心も特に男子で低かった｡4年制大学の進学を志望する学生にとっては, 3年制の多い医療技術系短期大学には関心が寄せがたく, 4年制へ移行した場合には関心も志望も高まる事が認められた｡将来この分野を担う人材の確保には現在の医療短大の4年制化や医療技術職の待遇改善が重要である と結論づけた｡A survey was carried out by senior high school students in Okayama and nine surrounding prefectures from July through September, 1996. Data obtained from 1998 twelfth-grade students studying general courses at 52 schools were computed. Eighty four percent of the 891 male students and 54.2% of the 1,077 female students wished to go on to senior college. Sixtyseven percent of the male students wished to enter science college courses, while only 25.8% of female students did. The rate of students who knew about the course of allied medical science in senior or junior colleges was 63.8% for male students and 82.5% for female students. The nursing course was popular among both sexes, but other courses such as radiological technology, medical technology, physical therapy and occupational therapy were not well known, especially among male students. It was thought that students wishing to go on to the senior college were not interested in those areas which were usually taught in three year courses at junior colleges or special (vocational) schools. Accordingly, the data showed that their interests in the allied medical course would be increased by the shifting it to a four year course. To attract senior high school students who are both intelligent and talented in the medical and health field, the authors concluded that the further improvement of educational course for allied medicine, especially such as shifting to a four year course, and the bettering of position of comedical workers through it are needed

Extracellular matrix components and soluble VCAM-l1n synovial fluid with osteoarthritis of the knee

Abstract It is generally accepted that the number of patients with osteoarthritis (OA) of the knee must increase in the future because of the increase of the aged. The pathogenic mechanisms of OA haven't been elucidated yet, but it is supposed that mediate pannus may cause the factors for irreversible cartilage damage in the inflammatory process. Therefore it is very important to find and predict OA at the early stage. Now there were many indices to determine joint damage with OA, for example, cytokines including IL, TNF, TGF, matrix metalloproteinase (MMPs), tissue inhibitor of metalloproteinase (TIMPs), proteoglycan (PG), hyaluronate (HA), super oxide, adhesion molecules including, ICAM-l, VCAM-I and PECAM. In this study, we measured that the soluble VCAM-l (sVCAM-I), soluble collagen (sCOLL), heparansulfate proteoglycan (HSPG), uronic acis and CRP in synovial fluids (SF) with OA. A positive correlation between the level of sVCAM-1 and sCOLL (p = 0.652) was noted. However there was no factor showing a correlation with CRP. It was suggested that the levels of sVCAM-I and sCOLL in the synovia fluid were active inflammation indices of knee damage, as the different aspects from CRP. The level of HSPG also showed a unique trend because HSPG on the cell surface was quickly digested endocytosis, so that it was not suitable for the inflammatory index in synovial fluid with OA