THE KINETICS AND MECHANICS OF CELL ADHESION MOLECULES

1NTRODUCTlONBindingviaadhesivereceptorsisessentialtomanycellularfunctions.Thekineticratesarecriticaltosuchinteractionssincetheseparametersdeterminehowrapidlycellsbindandhowlongtheyremainbound.Incontrasttosolub1emolecu1es,theassociationanddissociationofbondscross-bridgingtwoaPposingsurfacesusuaIlytakepIaceinthepresenceofforces,asoneofthebiologicalfunctionsoftl1eadhesionmoleculesistoprovidernechanicallinkagebetweenceIls.Physicalforcescaninfluencethebindinginteractionsofedhesivebonds.Assuch…     

淋巴滞留脑病模型大鼠内侧隔核-斜角带垂直支一氧化氮合酶神经元的改变

目的 :探讨淋巴滞瘤脑病模型大鼠内侧隔核 斜角带垂直支 (Septummedialis verticaldiagonalbandcomplex ,SM vDB)神经元一氧化氮合酶 (Nitricoxidesynthase,NOS)表达与空间参考记忆变化的相关性。方法 :用免疫细胞化学方法显示记忆获得后及淋巴滞留脑病期间 ,大鼠SM vDB的NOS阳性神经元 ,并进行定量分析。结果 :(1 )训练组阳性神经元数比对照组分别增加 96.91 % ,62 .96% (P <0 .0 1 ) ,与逃避潜伏期呈明显负相关 (SMr=-0 .7646,P <0 .0 1 ;vDBr=-0 .81 5 2 ,P <0 .0 1 ) ;细胞面积分别增加 42 .5 1 % ,3 5 .0 0 % (P <0 .0 1 ) ;免疫反应灰度值分别增加1 3 0 .5 1 % ,1 1 5 .41 % (P <0 .0 1 )。 (2 )脑病模型组与假手术组相比 ,阳性神经元数量分别减少 2 5 .91 % ,2 0 .1 3 % (P <0 .0 1 ) ,与逃避潜伏期呈明显负相关 (SMr=-0 .65 3 8,P <0 .0 1 ;vDBr=-0 .72 42 ,P <0 .0 1 ) ;细胞面积分别减少1 4.92 % ,1 2 .97% (P <0 .0 1 ) ,免疫反应灰度值分别减少 3 3 .3 4% ,41 .5 4% (P <0 .0 1 )。结论 :淋巴滞留性脑病神经元NOS表达减少 ,可能损害空间参考记忆的保持     

白细胞介素-1受体拮抗剂内含子2可变串联重复序列基因多态性与冠心病

目的　探讨白细胞介素 - 1受体拮抗剂内含子 2可变串联重复序列基因多态性与冠心病相关性 .方法　利用聚合酶链式反应技术对 86例冠心病及 10 0例正常对照者的白细胞介素 - 1受体拮抗剂基因进行扩增 .结果　冠心病患者IL - 1ra基因频率 :IL1RN1/ 10 .82 5 6 ,IL1RN1/ 2 0 .16 2 8,IL1RN1/ 30 .0 116 ,IL - 1ra等位基因频率 :IL1RN 10 .912 7,IL1RN 2 0 .0 814 ,IL1RN 30 .0 0 5 9,与对照组无差异 (p >0 .0 5 ) .结论　冠心病患者IL - 1ra基因多态性的分布与正常人无明显相关性     

团体咨询在大学生心理素质发展中的应用

目的　研究团体咨询对大学生心理素质发展的影响。方法　以团体活动的形式对 3 2名大学生进行为期 3个月的训练 ,训练前后分别运用 1 6PF和 SCL-90进行测量 ,并结合学生的成长报告进行训练效果分析。结果　经过 8次团体训练后 ,1 6PF问卷中有 4项人格因素有显著性改善 ,稳定性 ( 1 6.2 5± 5 .0 2 / 1 7.75± 4.1 6,P<0 .0 5 ) ,恃强性 ( 1 0 .91± 3 .82 /1 2 .3 4± 3 .65 ,P<0 .0 5 ) ,兴奋性 ( 1 3 .78± 4.0 7/ 1 5 .5 9± 3 .61 ,P<0 .0 1 ) ,敢为性 ( 1 2 .3 8± 3 .82 / 1 5 .0 6± 4.3 3 ,P<0 .0 0 1 ) ;SCL-90中人际因子发生显著变化 ,人际 ( 1 .81± 0 .5 5 / 1 .63± 0 .46,P<0 .0 5 )。结论　团体咨询对于促进成员的某些人格品质的发展 ,优化心理素质是有效的     

白细胞介素-1受体拮抗剂基因多态性与变态性鼻炎相关性探讨

目的　探讨白细胞介素 - 1受体拮抗剂 (IL -lra)基因多态性与变态性鼻炎相关性 .方法　利用聚合酶链式反应技术对 6 9例变态性鼻炎患者及 15 6例正常对照者的IL -lra基因进行扩增 .结果　变态性鼻炎患者IL - 1ra基因频率 :IL1RN1/ 10 .8116 ,IL1RN1/ 2 0 .1739,IL1RN1/ 30 .0 14 5 ,IL - 1ra等位基因频率 :,IL1RN 10 .90 5 8,IL1RN 20 .0 86 9,IL1RN 30 .0 0 72 ,与对照组无差异 (p >0 .0 5 ) .结论　IL - 1ra基因多态性的分布与变态性鼻炎无明显相关性     

The Improved Tensile Sensor in Vivo

1.IntroductionAtPresent,therearetwokindsofwaystomeasuretensileoftissue,theoneismeasurementinvitro,thiswayhasripetechnique,andeasyisthemeasurementinvivo,thiskindofmeasurementsystemhasnotbeenreporteduptonowindomestic,onlySalmon.Shaddonesomeworkatoverseas,buthisworkisnotperfectinbothdataprocessandprecision.So,thisresearch'spurposeisthatresearchandproduceanewmeasurementsystemoftensileinvivo.2.Designofsensor(1)TheapprovementofcrossbeamFig.1TheimprovedcrossbeamThenewbeamisadouble-layerbeam,therei…     

A double-element focused ultrasonic transducer used in hyperthermic oncology for superficial tumor

Inrece11tyears,ultrasOI1icl1ypotl1el'lnicOI1col0gyisdrawi11gn1oreattenti0nforitssafetyand1bcusingeffectiveIy.Intl1e7thRolnell1terl1atio11alfIype1'tl1ermicOl1coIogyConference,itwasviewedasaverypI-0misingheatingmethod.Ultrasonichyp0thermic0ncologyissimilarwithotherhyperthermaIInetl10dsforcancerinbasicprincipIes.Ultrasonicwaveenergy,afterbeingabs0rbedbythebodytissues-turnsi11toheatenergywl1icl1raisesthetissue'stemperature.Tumortissueslackblo0dandfuncti0npo0rIyincycli11g,sotheycaneasiIytobe…     

OBTAINING AND PR0CESSING OF DIFFRACTION ANDBACKSCATTERING IMAGE OF POLYDISPERSE RED BLOOD CELL

1.IntroductionAnewinstrumentisdeveIopedonthebasisofbackscatteringformeasuringredbloodcellsurfaceconcavity.Themethodoftheorderedarrangemelltofparticlesisdescribed,andthefiniteelement-ariificialtrAnsmittingboundarymethodisintroduced.2.PhysicalPrinciplesFraunhoferdiffractiontheorylp0intsoutthatthetotallightdiffiactionintensityisNtimesasmuchasthedimactionintensityofasingleredbIoodcel1,I(P,q)roNI(')(P,q).(l)FromHe-Nelasc.theincidentrayreachesthedilutepoolinFig.1.Thedidrictionandbackscattering…     

压宁定治疗术中高血压50例报告

我院自 1 997年 7月至今 ,用德国产的压宁定治疗 ,加大麻醉深度和施用止痛药物都难以控制的血压升高反应 50例 ,效果满意。1　临床资料和方法1 .1　一般资料50例中 ,男 30例 ,女 2 0例 ;年龄 4 6— 72岁 (平均 61 .5岁 )体重 4 9— 75ｋｇ(平均59ｋｇ) ,其中 1 5例患原发性高血压 ,全部患者都是经胸肺癌切除术 ,ＡＳＡ在Ⅱ级以上。1 .2　麻醉方法病人术前半小时用安定 1 0ｍｇ ,阿托品 0 .5ｍｇ ,诱导用硫喷妥钠 5— 8ｍｇ .ｋｇ 1 (平均 6.4ｍｇ .ｋｇ 1 )琥珀胆碱 2ｍｇ .ｋｇ- 1芬太尼 3ｕｇ·ｋｇ- 1,ｉｖ .维持用 1 %普鲁卡因 ,0 .0 1 …     

The Clinic Study of Reducing Vestibular Sensitivityby Electronic Stimulation on Papillae

1.IntroductionInthisarticle,aphysicaltreatmentonmotiongidinessisbroughtforwardandaspeciallydesignedelectronicstimulationdeviceisdevelopedwhichcouldgeneratetwokindsofelectronicstimulationbyaccelerationdirection.Thisdeviceisappliedonclinictests,theresultsdemonstratethatstimulationontwosidemastoidacupointsbyapropercurrentcouldreducevestibularsensitivityandremovemotiongidiness.2.Materialandmethod1.Material(1)20cases(femalef10male:10)whohavemotionsickness.AgesarebetweenZIand45.(2)Awhirlingmotions…     

Immunohistochemical demonstration of c-erbB-2 oncoprotein in gastric adenocarcinoma: comparison of cryostat and paraffin wax sections and effect of fixation.

AIMS--To investigate the effects of fixation on the immunohistochemical demonstration of c-erbB-2 oncoprotein using paraffin wax and cryostat sections; to compare c-erbB-2 expression in non-neoplastic and neoplastic gastric tissues. METHODS--Adjacent blocks of tumour and non-neoplastic tissue from four gastrectomy specimens were put into a panel of 10 fixatives including acetone, B5, Bouin's fluid, Carnoy's fluid, buffered formalin, formol dichromate, zinc formalin, 4% paraformaldehyde, periodate-lysine-paraformaldehyde (PLP) and periodate-lysine-paraformaldehyde-dichromate (PLPD) before embedding in paraffin wax for sectioning. Similar tissue blocks were snap frozen and cryostat sections were postfixed in these fixatives, either alone or in combination, before immunostaining. RESULTS--In paraffin wax embedded sections the best fixative was PLP, and in frozen tissues the best results were obtained after fixation of cryostat sections in buffered formalin followed by cold methanol and acetone. Applying these fixatives to samples from a further 16 gastrectomy specimens, strong membrane staining of c-erbB-2 protein was found in the tumour in eight of 16 cases (50%) using paraffin wax sections, and staining was stronger in the better differentiated carcinomas. For frozen tissues, positive membrane staining was found in all gastric adenocarcinomas, but differential staining intensity associated with tumour differentiation could not be detected. CONCLUSIONS--These results indicate that fixation and paraffin wax embedding affect the results of immunohistochemical demonstration of c-erbB-2 in gastric cancer. The choice of fixative is critical in the demonstration and evaluation of c-erbB-2 protein expression by immunohistochemistry in gastric carcinomas. Staining results also vary depending on whether frozen or paraffin wax embedded tissues are studied.     

Enzyme-labeled Antigen Method: Factors Influencing the Deterioration of Antigen-binding Activity of Specific Antibodies during Formalin Fixation and Paraffin Embedding

The enzyme-labeled antigen method is an immunohistochemical technique detecting plasma cells producing specific antibodies in tissue sections. The probe is an antigen labeled with an enzyme or biotin. This immunohistochemical technique is appliable to frozen sections of paraformaldehyde (PFA)-fixed tissues, but it has been difficult to apply it to formalin-fixed, paraffin-embedded (FFPE) sections. In the current study, factors inactivating the antibody reactivity during the process of preparing FFPE sections were investigated. Lymph nodes of rats immunized with horseradish peroxidase (HRP) or a mixture of keyhole limpet hemocyanin/ovalbumin/bovine serum albumin were employed as experimental models. Plasma cells producing specific antibodies, visualized with HRP (as an antigen with enzymatic activity) or biotinylated proteins in 4% PFA-fixed frozen sections, significantly decreased in unbuffered 10% formalin-fixed frozen sections. The positive cells were further decreased by paraffin embedding following formalin fixation. In paraffin-embedded sections fixed in precipitating fixatives such as ethanol and acetone and those prepared with the AMeX method, the antigen-binding reactivity of antibodies was preserved. Fixation in periodate-lysine-paraformaldehyde and Zamboni solution also kept the antigen-binding reactivity in paraffin to some extent. In conclusion, formalin fixation and paraffin embedding were major causes inactivating antibodies. Precipitating fixatives could retain the antigen-binding reactivity of antibodies in paraffin-embedded sections.     

A simplified method for the detection of maedi-visna virus RNA by in situ hybridization.

A simplified in situ hybridization method for the detection of maedi-visna virus (MVV) RNA in cultured cells using 35S-labelled DNA probes is described. The protocol currently used in this laboratory for the in situ detection of MVV RNA involves paraformaldehyde fixation followed by extensive cellular pretreatment prior to hybridization. It was found that substitution of paraformaldehyde fixation with brief acetone treatment and the removal of subsequent pretreatment steps gave a similar level of hybridization signal to that of our standard protocol. Acetone fixed, non-pretreated samples were used to develop a double labelling procedure in which immunocytochemistry and in situ hybridization were combined to allow the simultaneous detection of visna virus antigens and RNA within the same cell.     

Immunofluorescence assay for human immunodeficiency virus antibody: investigation of cell fixation for virus inactivation and antigen preservation.

Four cell fixation procedures were investigated for their abilities to inactivate human immunodeficiency virus (HIV) and preserve its antigenicity for antibody detection by immunofluorescence in MOLT-4-T4 cells. Air-dried cell smears were fixed in cold acetone, in acetone-methanol (1:1), in acetone-methanol (1:1) followed by 70% ethanol and then methanol, or in paraformaldehyde-acetone. Acetone alone did not inactivate cell-associated HIV, but the other three procedures did. HIV inactivation was achieved by storage of acetone-fixed cells at -70 degrees for 40 days. Antigenicity was measured by immunofluorescence assay titrations of selected human sera, a cerebrospinal fluid, and a gp41 monoclonal antibody. Acetone provided the best fixation as measured by fluorescence intensity and antibody titers. The other fixation methods all yielded weaker fluorescence signals and/or decreased titers. Acetone fixation and storage for 40 days at -70 degrees C provides safe and accurate immunofluorescence assay reagents.     

Immunocytochemical staining of progesterone receptor in paraffin sections of human breast cancers.

Monoclonal antibodies (mAB) against progesterone receptor (PR) and the peroxidase-antiperoxidase (PAP) method to visualize PR in paraffin sections from 68 human breast cancers were used. Ten mAB, which recognize human PR on frozen sections, were tested. Six could detect PR in paraffin sections, with Li 417, LET 456, and LET 126 giving the best results. LET 126 antibody was used for most further studies. The effects of fixation with picric acid-formaldehyde (PAF), buffered-formalin, or with Bouin solution were investigated; all fixation methods allowed PR immunolabeling, although PAF or buffered formalin usually gave the best results. Positive staining was seen in the nucleus of carcinoma cells. Variations in intensity and extent of immunoreactivity were observed in all sections and among different regions of the same specimen. These were probably related to the heterogeneity of the tumor cell population. Results were compared with the PR content in the respective tumor tissues, determined by steroid-binding assay, and with immunocytochemistry on frozen sections. It was shown that there were correlations between the immunocytochemical staining (positive or negative) and the steroid binding assay (80%) and between the immunocytochemical staining on paraffin sections and on frozen sections (78%). Weaker intensity and fewer number of PR-positive cells were found for paraffin-embedded tumors. Estrogen receptors were also detected on adjacent sections from the same paraffin-embedded tissues by use of monoclonal anti-ER antibodies (ERICA-kit[Abbott Laboratories, Chicago, IL]) and DNase pretreatment. In conclusion, this immunocytochemical method for detection of PR and ER on paraffin sections offers an alternative to frozen tissue. It allows histologic and immunocytochemical studies on the same sample and retrospective studies on stored tissue blocks.     

Comparative evaluation of unfixed and fixed human neutrophils for determination of antineutrophil cytoplasmic antibodies by indirect immunofluorescence.

BACKGROUND: Antineutrophil cytoplasmic antibodies (ANCAs) are found in the sera of patients with vasculitides and ulcerative colitis. Using indirect immunofluorescence on ethanol fixed neutrophils, ANCAs can be divided into two types: those that give a cytoplasmic staining pattern (C-ANCA) and those that give a perinuclear staining pattern (P-ANCA). Some studies have indicated that the perinuclear staining pattern might be an artefact of alcohol fixation. AIMS: To observe any changes seen in the ANCA staining pattern using indirect immunofluorescence on unfixed neutrophils or neutrophils that had been fixed by ethanol, acetone, or paraformaldehyde. In addition, the effects of the different fixation methods on the sensitivity of the indirect immunofluorescence test were evaluated. METHODS: Twenty one sera from patients with ulcerative colitis and 19 from healthy controls were studied. In addition, 17 sera from patients with vasculitides, including eight with proteinase 3 (PR 3) positive C-ANCA and nine with myeloperoxidase (MPO) positive P-ANCA were included in the study. Antineutrophil cytoplasmic antibodies were analysed by indirect immunofluorescence on unfixed neutrophils or cells fixed by ethanol, acetone, or paraformaldehyde. RESULTS: All the ulcerative colitis associated ANCA positive sera presented a peri-nuclear staining pattern on both unfixed and fixed cells. The acetone and paraformaldehyde fixations decreased the ulcerative colitis P-ANCA titres. Paraformaldehyde fixation also decreased the MPO positive P-ANCA titres. In indirect immunofluorescence, the staining patterns of all the eight PR 3 positive C-ANCA sera and eight of the nine MPO positive P-ANCA sera did not change, even if unfixed or fixed cells were used. CONCLUSIONS: The ANCA staining patterns are not affected by the fixation method used, and are the same as when unfixed neutrophils are used; this suggests that the P-ANCA pattern is not an artefact of alcohol fixation. Furthermore, this study confirms that on ethanol fixed neutrophils, the antigen of ulcerative colitis associated P-ANCA is better exposed than on acetone or paraformaldehyde fixed cells.     

Preparation and observation of fresh-frozen sections of the green fluorescent protein transgenic mouse head

Hard tissue decalcification can cause variation in the constituent protein characteristics. This paper describes a method of preparating of frozen mouse head sections so as to clearly observe the nature of the constituent proteins. Frozen sections of various green fluorescent protein (GFP) transgenic mouse heads were prepared using the film method developed by Kawamoto and Shimizu. This method made specimen dissection without decalcification possible, wherein GFP was clearly observed in an undamaged state. Conversely, using the same method with decalcification made GFP observation in the transgenic mouse head difficult. This new method is suitable for observing GFP marked cells, enabling us to follow the transplanted GFP marked cells within frozen head sections.     

Protocol for ultrarapid immunostaining of frozen sections

Rapid immunostaining of frozen sections within a tolerable time span would be very helpful for intraoperative diagnosis. A protocol was therefore established using the enhanced polymer one-step staining (EPOS) system (Dako) with antibodies against leucocyte common antigen (LCA), cytokeratin (CK), and anti-melanoma (MEL). Best results with reliable and specific immunostaining and a labelling intensity comparable to standard immunostaining protocols were achieved with fixation of samples in 100% acetone for 20 seconds (CK, LCA) or two minutes (MEL), followed by incubation of the primary antibody and development of the chromogen reaction with 3,3'diaminobenzidine (DAB) for three and five minutes at 37 degrees C, respectively. The total procedure takes only 12 minutes, thus enabling rapid immunostaining on intraoperative frozen sections. Apart from its use in tumour classification, this method is especially useful in detecting tumour cells in sentinel lymph nodes.     

Effects of different fixatives on staining patterns of four lectins in cultured microvascular endothelial cells

Fixatives have significant influence on lectin histochemistry of tissue sections; however, their roles in lectin fluorescent staining of cultured cells remain unclear. In this study, using cultured microvascular endothelial cells (MVECs) from rat intestinal mucosa, the effects of seven fixatives on the fluorescent staining patterns of four lectins were investigated. The results indicated that every fixative gave concanavalin A (Con A) and wheat germ agglutinin (WGA) strong positive staining in different patterns. When fixed with Zenker’s, periodate–lysine–paraformaldehyde (PLP), 2·5% glutaraldehyde (GA), and 4% paraformaldehyde (PFA) solution, the cell membranes and cytoplasms stained, and the fluorescence in the cell edges was brighter. Rossman’s solution, 95% ethanol, and acetone fixation caused cytoplasmic staining, while the fluorescence was weak and evanescent when using acetone. Dolichos biflorus agglutinin (DBA) and Ulex europaeus agglutinin I (UEA I) were stained; revealing very faint fluorescence in cytoplasms just when PLP and 2·5% GA solution were used. In conclusion, this study shows that fixatives have significant effects on the fluorescent staining of MVECs. Fixatives containing aldehydes and mercury chloride are better options for cell membrane glycans, and those containing ethanol for cytoplasmic ones. Acetone fixation is not suitable for lectin histochemistry.     

Double Fixation with Paraformaldehyde and Acetone for Optimal Immunocytochemical Demonstration of DNA Polymerase α

Abstract

Because the fixation procedure is a crucial step for immunocytochemical staining of DNA polymerase α, we sought an optimal fixation for demonstration of the enzyme using HeLa S3 cells. Double fixtion with paraformaldehyde and acetone produced intense stainability when slides were dipped in a 4% paraformaldehyde solution for 10 min, then in acetone for 5 rnin at 4°C. In contrast, fixation with ethanol or methanol resulted in considerable reduction of immunoreactivity. Single fixation with paraformaldehyde or acetone yielded intermediate stainability. Poor stainability also resulted in decrease of positive cells. (The J Histotechnol 12:285, 1989)