蛙皮素对PC-3细胞骨架及细胞内游离Ca~(2+)的影响

目的:观察蛙皮素(BBS)对前列腺癌PC-3细胞骨架形态及细胞内游离Ca2+([Ca2+]i)浓度的影响。方法:①利用免疫荧光法(IH),结合激光扫描共聚焦显微镜(LSCM)检测10-5mol/L浓度BBS处理的PC-3细胞角蛋白(CK)的表达,以反映其对细胞骨架形态的影响;②应用Fluo-3/AM荧光标记技术和LSCM检测不同浓度BBS(10-9、10-7、10-5mol/L)处理的PC-3细胞[Ca2+]i浓度。结果:①10-5mol/L浓度的BBS可促进PC-3细胞CK表达及伪足形成;②BBS可提高PC-3细胞[Ca2+]i浓度,并具有浓度依赖性。结论:实验证明一定浓度的BBS可明显提高PC-3细胞[Ca2+]i浓度及CK表达,进而影响PC-3细胞骨架形态。本研究为探索BBS应用于肿瘤研究以及BBS作用后细胞内信息传递途径提供了基础。     

HBeAg时间分辨荧光免疫分析法的建立

采用平衡饱和法建立了乙型肝炎病毒e抗原(HBeAg)时间分辨荧光免疫分析法.以针对HBeAg的单克隆抗体G8包被板,双功能螯合剂异氰酸苄基二乙烯三胺四乙酸络合Eu3+及标记C4单抗,发光增强系统为以β-二酮体为主的增强液.数据采用Log-Logit法函数和四参数Logitc函数数据处理程序处理.结果表明方法的批内和批间CV分别为2.39%和5.28%,平均回收率为97.62%,灵敏度为0.58NCU/mL,可测范围为12.01-529.84NCU/mL,ED20、ED50和ED80分别为6.36NCU/mL、26.85NCU/mL和136.7NCU/mL.本方法与HBsAg有13.1%的交叉反应.Eu3+标记抗体-30℃保存6个月免疫反应性基本无损失,同批试剂连续5个月应用分析结果稳定.HBeAg时间分辨荧光免疫分析的质量参数优于EIA和IRMA.     

The role of phospholipase A2 in human acute pancreatitis

Summary Several studies suggest that the activation of pancreatic phospholipase A₂ (PLA₂) and its release from injured acinar cells play an important role in the pathogenesis of acute pancreatitis. Elevated catalytic activity of PLA₂ in serum is associated especially with severe forms of the disease. PLA₂ has been purified from human cadaver pancreas and an antiserum raised against the enzyme in rabbits. Immuno-histochemical localization of PLA₂ in pancreatic tissue was abnormal in acute pancreatitis. A time-resolved fluoroimmunoassay for human pancreatic PLA₂ has been developed. Increased serum concentrations of immunoreactive PLA₂ were found in acute pancreatitis during the first week after hospital admission. The values returned to normal somewhat more slowly than corresponding serum amylase values. The immunochemical determination of PLA₂ in serum provides a fast and specific detection of injury to pancreatic acinar cells. The pancreas is not the only source of PLA₂ in acute pancreatitis. The nonpancreatic PLA₂ may originate from various inflammatory cells, but this hypothesis remains to be proven.     

Time-resolved fluorometric assay for the detection of endostatin in chromatographically separated extracts of natural peptides

We present a heterogeneous non-competitive immunological detection assay for peptide and protein antigens from crude extracts of biological sources. This time-resolved fluoroimmunoassay (TR-FIA) has been designed in a solid-phase mode using 96-well microtiter plates. Using the rare-earth metal europium as a fluorescent marker, a highly sensitive, selective and efficient procedure was developed. This technique prevents from interferences of intrinsic protein fluorescence which is highly important for antigen measurement in complex matrices. The TR-FIA has been applied for the detection of circulating forms of the potential anti-tumor agent endostatin, a C-terminal fragment of collagen XVIII, and its close homolog collagen XV (restin) from hemofiltrate. Endostatin was detected with a limit of detection of 3 ng (150 fmol/well) and a broad dynamic range from 10-1000 ng/well.     

鼠抗人单克隆抗体LH7：2在营养不良型大疱性表皮松解症诊断中的应用

目的 探讨鼠抗人单克隆抗体LH7：2诊断营养不良型大疱性表皮松解症的意义。方法 采用间接免疫荧光法对临床诊断为营养不良型大疱性表皮松解症（DEB）,分别来源于3个不同家系各1名患者,包括2名显性遗传型营养不良型大疱性表皮松解症（dominant DEB,DDEB)和1名隐性遗传型营养不良型大疱性表皮松解症（recessive DEB,RDEB)进行皮肤基底膜带（BMZ）Ⅶ型胶原检测,同时选择单纯型大疱性表皮松解症（EBS）、大疱性类天疱疮及正常人各1名做对照。结果 3名对照BMZ免疫荧光阳性,2名DDEB荧光均减弱,RDEB荧光阴性。结论 鼠抗人单克隆抗体LH7：2诊断营养不良性大疱性表皮松解症有重要意义。     

Time-resolved confocal fluorescence microscopy of porphyrins for phototherapy

The application of a novel time-resolved confocal fluorescence microspectrometer to studies of the distribution and speciation of porphyrin photosensitizers in rat C6 cerebral glioma cells is described. The instrument combines a mode-locked argon ion laser excitation source with time-correlated single photon counting fluorescence detection and has sub-micron spatial and sub-nanosecond temporal resolution. The porphyrins studied were haematoporphyrin derivative (HpD), haematoporphyrin IX (HP), porphyrinc (Pc) and the tetrakiscarborane carboxylate ester of 2,4-(,-dihydroxyethyl) deuteroporphyrin IX (BOPP). From the heterogeneous emission observed in vitro, assignments and spatial location of various porphyrin species are proposed.     

A Time-Resolved Fluorescence Immunoassay (DELFIA) Increases the Sensitivity of Antigen-Driven Cytokine Detection

Abstract

In an effort to improve the quantification of the low levels of cytokines released in response to antigenic stimulation of T cells, a sandwich dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA) was developed and compared to a standard sandwich ELISA. The DELFIA enhanced the sensitivity of a mouse IL-2 assay 8- to 27-fold, and a human GM-CSF assay 10-fold, as compared to colorimetric ELISA. The increase in sensitivity allows for the use of lower sample volumes per well, and the ability to run more assays per supernatant sample. This sensitive, nonisotopic alternative to other cytokine detection methods will be useful for those researchers wanting to quantitate low levels of antigen-driven cytokine production.     

时间分辨荧光免疫分析法在乙型肝炎病毒表面抗原检测中的应用

目的探讨时间分辨荧光免疫分析法(TRFIA)定量检测乙型肝炎表面抗原(HBsAg)的临床应用价值。方法分别用TRFIA和酶联免疫吸附试验(ELISA)检测237例血清样本HBsAg,并在高、中、低三浓度中各取20例标本用两种方法复检,对比结果进行分析。结果在237例标本中,TRFIA测得阳性99例,ELISA阳性91例,阳性率差异无统计学意义(P>0.05);高、中浓度标本各20例均为阳性,检出率差异无统计学意义(P>0.05),而低浓度标本中用TRFIA测得阳性20例,ELISA只有12例阳性,检出率差异有统计意义(P<0.01)。结论 TRFIA较敏感,在低浓度HBsAg检测中更具优势,提高了临床检测水平,具有较高的实用价值。     

4D microstructural changes in dentinal tubules during acid demineralisation

ObjectiveDental erosion is a common oral condition caused by chronic exposure to acids from intrinsic/extrinsic sources. Repeated acid exposure can lead to the irreversible loss of dental hard tissues (enamel, dentine, cementum). Dentine can become exposed to acid following severe enamel erosion, crown fracture, or gingival recession. Causing hypersensitivity, poor aesthetics, and potential pulp involvement. Improving treatments that can restore the structural integrity and aesthetics are therefore highly desirable. Such developments require a good understanding of how acid demineralisation progresses where relatively little is known in terms of intertubular dentine (ITD) and peritubular dentine (PTD) microstructure. To obtain further insight, this study proposes a new in vitro method for performing demineralisation studies of dentine.MethodsAdvanced high-speed synchrotron X-ray microtomography (SXM), with high spatial (0.325 μm) and temporal (15 min) resolution, was used to conduct the first in vitro, time-resolved 3D (4D) study of the microstructural changes in the ITD and PTD phases of human dentine samples (∼0.8 × 0.8 × 5 mm) during 6 h of continuous acid exposure.ResultsDifferent demineralisation rates of ITD (1.79 μm/min) and PTD (1.94 μm/min) and their progressive width-depth profiles were quantified, which provide insight for understanding the mechanisms of dentine demineralisation.SignificanceInsights obtained from morphological characterisations and the demineralisation process of ITD and PTD during acid demineralisation would help understand the demineralisation process and potentially aid in developing new therapeutic dentine treatments. This method enables continuous examination of relatively large volumes of dentine during demineralisation and also demonstrates the potential for studying the remineralisation process of proposed therapeutic dentine treatments.     

Subtraction MR Venography Acquired from Time-Resolved Contrast-Enhanced MR Angiography: Comparison with Phase-Contrast MR Venography and Single-Phase Contrast-Enhanced MR Venography

ObjectiveTo evaluate the image characteristics of subtraction magnetic resonance venography (SMRV) from time-resolved contrast-enhanced MR angiography (TRMRA) compared with phase-contrast MR venography (PCMRV) and single-phase contrast-enhanced MR venography (CEMRV).ResultsSingle-phase contrast-enhanced MR venography showed better image quality (median score 4 in both reviewers) than did the other two MRVs (p < 0.001), whereas SMRV (median score 3 in both reviewers) and PCMRV (median score 3 in both reviewers) had similar image quality (p ≥ 0.951). SMRV (median score 0 in both reviewers) suppressed arterial signal better than did the other MRVs (median score 1 in CEMRV, median score 2 in PCMRV, both reviewers) (p < 0.001). The dural sinus score of SMRV (median and interquartile range [IQR] 48, 43-50 for reviewer 1, 47, 43-49 for reviewer 2) was significantly higher than for PCMRV (median and IQR 31, 25-34 for reviewer 1, 30, 23-32 for reviewer 2) (p < 0.01) and did not differ from that of CEMRV (median and IQR 50, 47-52 for reviewer 1, 49, 45-51 for reviewer 2) (p = 0.146 in reviewer 1 and 0.123 in reviewer 2). The SNR and CNR of SMRV (median and IQR 104.5, 83.1-121.2 and 104.1, 74.9-120.5, respectively) were between those of CEMRV (median and IQR 150.3, 111-182.6 and 148.4, 108-178.2) and PCMRV (median and IQR 59.4, 49.2-74.9 and 53.6, 43.8-69.2).ConclusionSubtraction magnetic resonance venography is a promising MRV method, with acceptable image quality and good arterial suppression.