树突细胞联合细胞因子诱导的杀伤细胞对急性白血病细胞的体外杀伤作用

目的:研究细胞因子诱导的杀伤细胞(CIK)与同源树突状细胞(DC)共培养后CIK细胞的表型、增殖活性的变化,及抗急性白血病细胞活性.方法:正常人外周血单个核细胞诱导DC和CIK细胞,将DC与CIK共培养,以CIK细胞单独培养为对照.用台盼蓝活细胞计数计算细胞扩增倍数,MTT法测定杀伤活性,流式细胞术分析免疫表型.结果:DC-CIK细胞增殖能力明显高于CIK细胞(P＜0.05); DC、CIK细胞共培养后,CD3+ CD8+、CD3+ CD56+双阳性细胞比率较同条件下CIK细胞组显著增多(P＜0.05);在2.5∶1-20∶1的效靶比范围内,DC-CIK共培养物对AML细胞的杀伤率显著高于CIK细胞(P＜0.05),且杀伤率与效靶比呈正相关.结论:DC-CIK细胞的增殖能力、对AML细胞的杀伤活性均高于CIK细胞,为DC-CIK细胞免疫治疗提供了实验和理论依据.     

免疫脂质体与人胃癌细胞的相互作用

我们把抗人胃癌细胞M85的单克隆抗体3Hll插入脂质体的脂双层做成免疫脂质体,研究了这些免疫脂质体与M85细胞的相互作用.结果表明,M85细胞对免疫脂质体的吸收与温育时间、温度密切相关.在37℃温育10小时,细胞吸收的免疫脂质体的量是在4℃时的7倍左右.37℃温育5小时,靶向脂质体结合在M85靶细胞上的量是非靶向脂质体的2倍多;与非靶细胞—人皮肤成纤维细胞Fb32的结合量只有与M85靶细胞结合量的1/6.游离单抗3Hll能够抑制靶向脂质体与靶细胞的结合,而专一性与3Hll抗体不同的单抗3G9则不能.NH_4Cl和NaN_3等内吞作用抑制剂使靶细胞吸收免疫脂质体的量分别减少58%和79%.与此同时,NH_4CI还能抑制包入脂质体的ADM,而不能抑制游离ADM的细胞毒作用.因此我们的结论是,3Hll免疫脂质体能够与靶细胞专一地结合,并把所荷载的物质主要经内吞作用输送到细胞内,杀伤靶细胞.用荧光显微镜对与荧光免疫脂质体温育的M85细胞进行的形态观察也支持上述结论.     

金鱼表皮的带突起细胞的扫描电镜观察

本文对60只金鱼的皮肤进行了扫描电镜研究。除观察了带有突起的诸种细胞外,尚在雄性金鱼的头盖部皮肤发现一种从未报道过的新型细胞突起,呈指状,约40×20μm。其形态大小与性结节完全不同。经喂食11-酮睾丸酮后,该突起增至近乎原来的两倍。其机能是否与性机能有关尚待进一步探讨。     

靶向树突状细胞表面分子DEC-205的长循环免疫脂质体稳定性影响因素的数学模型

在多相分散体系的动力稳定性理论基础上,设计了以DEC-205单抗为导向、以脂质体为载体、DCs为靶点的免疫策略,将包裹药物的脂质体特异性地靶向树突状细胞（DCs）．对影响DEC-205的免疫脂质体稳定性的各种因素如粒径分布等进行考察分析,对条件进行优化,构建了经DEC-205单抗靶向DCs的免疫脂质体模型．模型的成功构建为进一步研究抗原靶向DEC-205受体后的体内免疫应答情况提供了工作基础,有望开发一种新型DCs疫苗应用于临床．     

生长抑素对白血病细胞增殖影响的免疫组织化学研究

目的　研究生长抑素 (somatostatin ,SS)对人红白血病K5 6 2细胞和白血病患者骨髓单个核细胞 (BMM NC)增殖的影响。方法　将 8肽生长抑素 (奥曲肽 ,octreotide ,SMS)在体外作用于K5 6 2细胞和BMMNC ;用原位末端标记 (TUNEL)技术、免疫组织化学方法在光镜下随机计数阳性细胞数 ,并用图象分析系统分析bcl 2蛋白和CD34 抗原表达的光密度值 (OD值 )。结果　与对照组比较 ,用SMS 10 -6mol/L处理的K5 6 2细胞组TUNEL检测凋亡细胞明显增多(P <0 0 1)。在加SMS 10 -8mol/L～ 10 -6mol/L处理的白血病患者BMMNC组 ,bcl 2蛋白的表达与未加SMS组比较 ,阳性细胞数明显减少 (P <0 0 1) ;OD值P <0 0 1。CD34 抗原表达在SMS10 -6mol/L和 10 -7mol/L处理的白血病患者BMMNC组 ,阳性细胞数比与未处理组明显减少 (P <0 0 1) ;OD值明显低于对照组 ,分别为P <0 0 1和P <0 0 5。结论　SMS可抑制K5 6 2细胞和白血病患者BMMNC增殖 ,诱导凋亡 ;减少bcl 2蛋白和CD34 抗原的表达 ;SMS对白血病细胞具有抑制增殖的作用。     

掺入胆固醇和阴离子磷脂对阿霉素免疫脂质体性能的影响

本文研究了胆固醇和阴离子磷脂的掺入对阿霉素—磷脂酰乙醇胺免疫脂质体包裹效力及其在PBS缓俄冲液和在50%血清中的稳定性的影响,并对这种影响可能的机理进行了讨论.阴离子磷脂PG和DPG的掺入使ADM免疫脂质体包裹ADM的效力大大增加,使ADM与PE的克分子比从(0.08—0.8)%增加到75%,其中DPG比PG更有效.掺入胆固醇可以明显提高ADM脂质体在缓冲液和在血清中的稳定性,但使脂质体包裹ADM的效力下降.通过脂质的选择和制备条件的摸索,我们成功地用超声法制备了包裹抗癌药(ADM),表面带有抗人胃癌细胞M85单克隆抗体3HII的小单层脂质体(SUV),其起脂质份是PE:PG:Chol:ADM(4:2:2:1),脂质体内的ADM与PE的克分子比为17—25%.经电镜观察,脂质体直径在60—80nm范围内,大以比较均一.这种脂质体在PBS缓冲液中于室温下保存12天,仍然能够保持其包裹药的70%;在50%的血清中,37℃1小时,能够保持其包裹药的80%.     

靶向树突状细胞表面分子DEC-205长循环免疫脂质体稳定性模型的构建及其生物学特性

应用多相分散体系的动力稳定性和聚结稳定性理论,以薄膜分散法构建了靶向树突状细胞（dendritic cells,DCs）表面分子DEC-205长循环免疫脂质体（anti—DEC-205 iLPSM）的稳定性模型,并对其物理稳定性、生物学特性等进行了考察。结果表明经优化后的脂质体4℃贮存7d后粒径分布变化较小;FTTC-dextran累积泄漏率小于7％;耦联抗DEC-205的免疫脂质体（anti—DEC-205 iLPSM）可特异性地识别DCs,并作为良好载体将FITC-dextran带入DCs浆内。anti—DEC-205 iLPSM模型的构建为进一步研究抗原靶向DEC-205受体后的体内免疫应答情况提供了工作基础,有望开发一种新型DCs疫苗应用于临床。     

Scanning Electron Microscopy of Schizogony in Eimeria tenella

SYNOPSIS. Developing 2nd- and 3rd-generation schizonts of Eimeria tenella were found in the ceca of chicks infected orally with sporulated oocysts. Several free 2nd-generation schizonts, which varied in diameter from 11 to 21.6 μm, were found on the epithelial surface of the cecum. Some schizonts appeared to have lost merozoites. Other schizonts were intact, one of which was surrounded by an unbroken membrane that followed the contours of the merozoites. Third-generation schizonts, much smaller than 2nd-generation schizonts and with fewer merozoites, were found only on cut or fractured surfaces of the cecal tissue. Third-generation merozoites appeared shorter and thicker than those of the 2nd-generation and were attached to the schizont residuum. A form with conical protuberances and another with 4 triangular segments were found; they were believed to be developing stages 3rd-generation schizonts.     

Three-Dimensional Structure of Macronuclei from Paramecium Determined by Scanning Electron Microscopy*

SYNOPSIS. Macronuclei of Paramecium primaurelia were isolated and examined by scanning electron microscopy. These nuclei consisted of a closely packed array of chromatin bodies measuring ～ 0.2 μm in diameter. We estimated there were ～ 30,000 such bodies/macronucleus, 20 times more than the number of unit genome equivalents. This suggests that a unit genome is physically shared by several chromatin bodies.     

The Dimension of Trichomonas vaginalis as Measured by Scanning Electron Microscopy

It is known that physicochemical conditions (e.g., pH, temperature, and ionic strength) affect the size of trichomonads. In this study, the sizes of 4 isolates of Trichomonas vaginalis cultured for more than a year (called "old T") and 3 isolates freshly isolated from vaginitis cases (called "fresh T") were compared by scanning electron microscopy. Although the fresh T had shorter body length, body width, and flagellar length than old T, total length (about 26 µm), including body length, flagella length, and axostyle length was almost the same in the 2 groups. A striking difference was observed between the axostyles of the 2 groups; the axostyle length of the fresh T (8.2 µm) was more than twice as long as that of the old T (4.0 µm). However, in several parasitology textbooks, the length of T. vaginalis is said to vary widely from 7 to 32 µm, and its undulating membrane is said to extend about half way (53.5%) to the posterior end of the body. On the other hand, in our study, the undulating membrane was observed to extend more than 3/4 of the body length (72.1%) in old T, whereas in fresh T it could not be measured. Taken together, we suggest that T. vaginalis averages 26 (21-32) µm in total length, with 9.5 (7.4-11.4) µm of body length and 6.8 (5.3-7.7) µm of width, and its undulating membrane extending 3/4 of its body length. Therefore, these findings may provide useful information for morphological characteristics of T. vaginalis.     

Operando Observations of SEI Film Evolution by Mass‐Sensitive Scanning Transmission Electron Microscopy

The solid electrolyte interphase (SEI) spontaneously formed on anode surfaces as a passivation layer plays a critical role in the lithium dissolution and deposition upon discharge/charge in lithium ion batteries and lithium‐metal batteries. The formation kinetics and failure of the SEI films are the key factors determining the safety, power capability, and cycle life of lithium ion and lithium‐metal batteries. Since SEI films evolve with the volumetric and interfacial changes of anodes, it is technically challenging in experimental study of SEI kinetics. Here operando observations are reported of SEI formation, growth, and failure at a high current density by utilizing a mass‐sensitive Cs‐corrected scanning transmission electron microscopy. The sub‐nano‐scale observations reveal a bilayer hybrid structure of SEI films and demonstrate the radical assisted SEI growth after the SEI thickness beyond the electron tunneling regime. The failure of SEI films is associated with rapid dissolution of inorganic layers when they directly contact with the electrolyte in broken SEI films. The initiation of cracks in SEI films is caused by heterogeneous volume changes of the electrodes during delithiation. These microscopic insights have important implications in understanding SEI kinetics and in developing high‐performance anodes with the formation of robust SEI films.     

Scanning Electron Microscopy of Budding and Metamorphosis in Discophrya collini (Root)*

SYNOPSIS. Budding and metamorphosis in the suctorian ciliate, Discophrya collini, have been investigated by scanning electron microscopy. The adult body form, tentacles, stalk, and attachment disk are described. A field of depressions or small pits was observed in the pellicle of adult suctorians in the early stages of bud formation. These pits deepen and coalesce until one large pore, the birth pore, remains. Cilia protrude through the pore, and as eversion of the bud proceeds the meridional arrangement of the larval ciliation is evident. After eversion is completed, a pronounced division furrow is found between the adult and soon-to-be-released swarmer. The stalk-forming region is seen on swarmers. Metamorphosing swarmers produce tentacles upon settling before any indication of ciliary resorption. Resorption of cilia and change in body form occur progressively with the production of the attachment disk and stalk.     

Scanning Electron Microscopy of Stomatogenesis Accompanying Conjugation in Euplotes aediculatus*

SYNOPSIS. The succession of morphologic changes in the feeding apparatus (peristome) accompanying conjugation and postconjugant development in the hypotrich Euplotes aediculatus has been examined by scanning electron microscopy (SEM). The details of stomatogenesis inferred from earlier light-microscopic studies of silver-stained preparations have been confirmed and extended. The elaborate peristome is the dominant surface feature of vegetative Euplotes. In conjugation, the ciliates are joined in their peristomial regions; as the conjugants separate, the old feeding apparatus is seen to be disrupted and partially resorbed. In its place is the crescent-shaped primordium of a new peristome, which develops as part of a general cortical reorganization. This primordium expands anteriorly, unfurling a new crown of ciliary membranelles that soon replaces the remaining preconjugant membranellar band. The resulting “exconjugant peristome'’is characterized by a greatly reduced number of adoral membranelles and the absence of paroral membranelles, buccal cavity, and cytostome. Exconjugants thus cannot feed for 2–3 days, until the missing peristomial components are replaced. This occurs by means of a 2nd cortical reorganization, during which new membranelles, developing from another peristomial rudiment, are added directly to the abbreviated exconjugant set. A new buccal cavity is concurrently sculpted as the primordial depression enlarges, and the cells can resume feeding sometime during the 4th day after separation. The implications of this mode of stomatogenesis and the nonfeeding condition are discussed, as are the advantages of SEM for studies of ciliate morphogenesis.     

A Simple Drying Method for Field Emission Scanning Electron Microscopy for Chromosomes

Hexamethyldisilazane treatment and subsequent air drying of spread plant chromosomes is compared with critical point drying. The two procedures are equivalent for preparing chromosomes for examination by field emission scanning electron microscopy at low voltage.     

Scanning Electron Microscopy of Tintinnopsis parva: Studies on Particle Accumulation and the Striae*

SYNOPSIS. Lorica building was studied experimentally for Tintinnopsis parva Merkle, an agglutinated tintinnid. This species incorporated either siliceous or Ca-rich particles. Evidence of agglutination was seen within 30 min after the addition of particles to cultures of the protozoa. Fully agglutinated loricae were produced by the proter; partially incrusted loricae attributed to an opisthe were also recovered. The cytoplasmic evaginations of unknown function called striae were seen in the scanning electron microscope in better relief than ever before. They may provide the mechanism whereby the enclosed capsules are brought into contact with prey organisms.     

Light and Scanning Electron Microscopy of Greenbug Aphid Damage in Wheat Using the Same Section

A method has been developed to enable correlative light microscopy (LM) and scanning electron microscopy (SEM) on the same section of wheat (Triticum aestivum L.) leaves infested by greenbug aphids (Schizaphis gra-minum Rondani). Segments of infested leaf tissue were fixed, embedded in paraffin, sectioned, and affixed to slides by standard histological techniques. Serial sections were viewed by LM as temporary mounts in xylene. Sections of interest were identified and re-embedded in fingernail polish, affixed to aluminum stubs, freed of polish with ethyl acetate or acetone, and sputter-coated for SEM. SEM of re-embedded leaf sections showed excellent preservation of leaf anatomy. The same aphid tracks and regions of cell damage identified by LM were visible. SEM increased resolution and provided a much clearer sense of the three-dimensional relations involved in the interaction between plant and insect.     

Light and Scanning Electron Microscopy of Greenbug Aphid Damage in Wheat Using the Same Section

A method has been developed to enable correlative light microscopy (LM) and scanning electron microscopy (SEM) on the same section of wheat (Triticum aestivum L.) leaves infested by greenbug aphids (Schizaphis gra-minum Rondani). Segments of infested leaf tissue were fixed, embedded in paraffin, sectioned, and affixed to slides by standard histological techniques. Serial sections were viewed by LM as temporary mounts in xylene. Sections of interest were identified and re-embedded in fingernail polish, affixed to aluminum stubs, freed of polish with ethyl acetate or acetone, and sputter-coated for SEM. SEM of re-embedded leaf sections showed excellent preservation of leaf anatomy. The same aphid tracks and regions of cell damage identified by LM were visible. SEM increased resolution and provided a much clearer sense of the three-dimensional relations involved in the interaction between plant and insect.