Preparation and identification of multifunctional mesoporous silica nanoparticles for in vitro and in vivo dual-mode imaging,theranostics, and targeted tracking

Mesoporous silica nanoparticles (MSNs) can provide a structural foundation for a new generation of nanocarriers with a broad range of functionalities. Multifunctional MSNs can serve as all-in-one diagnostic and therapeutic tools that can be used to simultaneously visualize and treat various diseases, such as cancer. This research study is the first time that two lanthanide-based imaging systems have been combined to incorporate controlled drug release and targeted tracing into a single MSN-based nano-platform for a novel theranostic drug delivery system. Doping lanthanide ions, i.e., europium (Eu) and gadolinium (Gd) ions, into an MSN structure (EuGd-MSNs) imparts fluorescence and magnetism to the nanostructure that can be used to develop magnetic resonance imaging (MRI) and biological fluorescence tools. Current cancer research has revealed that most human cancer cells express a large number of folate receptors on their surface. Grafting folic acid (FA) onto the EuGd-MSN surface (EuGd-FA-MSNs) imparts a targeting function to the MSN because of the specificity of the binding of FA to cell surface receptors. Furthermore, grafting anticancer drugs, such as camptothecin (CPT), onto the surface of these MSNs by forming disulfide bonds (EuGd-SS-CPT-FA-MSNs) enables intracellular controlled drug release. A high concentration of intracellular glutathione cleaves the disulfide bond to release the drug and treat the disease. The results of in vitro and in vivo studies show that the functionalized MSNs can be successfully used as a platform to integrate dual-imaging, targeting, and therapeutic treatment in multifunctional diagnosis drug delivery systems.     

荧光原位杂交法快速鉴定血培养阳性葡萄球菌的临床应用

目的快速鉴定血培养中的金黄色葡萄球菌和凝固酶阴性葡萄球菌(CoNS),结合临床快速判定是否为污染菌。方法采用荧光原位杂交法鉴定血培养中的金黄色葡萄球菌和CoNS,杂交结果若为CoNS,根据临床资料进行判断,并与文献推荐的污染判断法进行结果比较。结果探针的特异性经由标准菌株和临床分离菌株证实。金黄色葡萄球菌探针的特异性和敏感性均为100%,GoNS探针的特异性和敏感性分别为100%和95.5%。179株CoNS中117株判断为污染菌,污染率为68%,与文献推荐的污染判断方法一致。结论荧光原位杂交法适用于血培养中的金黄色葡萄球菌和CoNS的快速鉴定,以排除CoNS污染。     

应用荧光多重聚合酶链反应法检测结直肠癌微卫星不稳定

目的建立一种敏感、稳定、高通量的结直肠癌微卫星不稳定(MSI)检测技术。方法105例散发性结直肠癌患者取其新鲜癌组织及正常肠黏膜,DNA提取,荧光多重聚合酶链反应(PCR)扩增5个微卫星位点,应用GeneScan方法分析PCR产物。结果105例结直肠癌患者26例(24.7%)存在微卫星不稳定,其中MSI-H 14例(13.3%),MSI-L 12例(11.4%),MSS 79例(75.3%)。D5S346、BAT26、BAT25、D17S250、D2S123突变率分别为5.6%、8.6%、10.5%、8.6%、10.5%。结论应用荧光多重PCR可以高通量检测结直肠癌微卫星不稳定,敏感性高,结果可靠,适合临床应用。     

On the physical properties of red cell ghost membranes in the affective disorders and psychoses A fluorescence polarization study

Membrane order was measured in the erythrocyte ghost membranes of DSM-III schizophreniform disorder (SF), DSM-III schizophrenic (SCZ) and DSM-III manic (bipolar) (M) patients and a group of age- and sex-matched controls. Fluorescence polarization with the probe 1,6-diphenyl1-1,3,5-hexatriene was used to determine the steady-state fluorescence anisotropy (r_s). The SF group showed a significant increase in r_s (Δr_s = 0.037) from the control group. Although the means were not significantly different, 3 of 8 Ms and 5 of 8 SCZs also had r_s values > the highest control value. Thermotropic behavior of the membranes was evaluated over the range of 40 to 20°C. No difference among groups in membrane enthalpy was detected. Thus, the differences in r_s appear to be associated with differences in entropy. Phosphatidylcholine (PC) levels, which were known to be abnormal in these patients, were compared with the r_s values. A significant (P < 0.001, R= -0.63) linear correlation between r_s and membrane PC levels was observed. Overall these data further support the view that unusual membrane biophysical factors may occur with high frequency in the psychoses and affective disorders.     

荧光分光光度法检测纳米粒运载的阿霉素在大鼠体内的分布

目的　观察由半乳糖化磁性白蛋白纳米粒运载的阿霉素经舌静脉给药后在大鼠体内的的分布状况。方法　全部大鼠随机分为 4组 ,经舌静脉 ,按分组分别注射 :游离阿霉素组(FADM ) ;磁性阿霉素白蛋白纳米粒 (MADM ) ;半乳糖化磁性阿霉素白蛋白纳米粒组 (Gal2 9 ADM ) ;半乳糖磁性阿霉素白蛋白纳米粒 外加磁场 (Gal2 9 ADM M ) ,剂量均为阿霉素 2 .5mg/kg体重。取全血、心、肺、肝、脾、肾。全血制成血浆 ,器官组织制成匀浆 ,盐酸乙醇法提取阿霉素 ,用荧光光度计测量。结果　静脉注射同等剂量、不同剂型的阿霉素药物后 ,阿霉素在器官中的蓄积程度从高到低 :肝 :D靶肝 >D非靶肝、C >B >A ;心脏、肾、血浆 :A >B >D、C ;脾 :B >A、C、D ;肺 :B>A >C、D。磁性阿霉素白蛋白纳米粒注入体内后 ,在心、肺、肝、脾、肾中的药物浓度在 15～ 3 0min达峰值 ,而半乳糖化后 ,阿霉素的药物峰值提前到 5min或之前。外加磁场和未加磁场的半乳糖化磁性阿霉素白蛋白纳米粒组的药物靶向指数和药物选择指数是均高于磁性白蛋白纳米粒。结论　磁性阿霉素白蛋白纳米粒经半乳糖化后 ,可显著增强阿霉素对肝脏的靶向性 ,并显著降低心、肺、脾、肾、血浆肝外器官的组织阿霉素浓度。利用外加磁场 ,可提高阿霉素在肝脏特定部位蓄积的能力。     

早期糖尿病视网膜病变的视网膜电图分析

目的 分析早期糖尿病视网膜病变(DR)的闪光视网膜电图(F—ERG)和视网膜电图震荡电位(OPs)各参数的变化特点，寻找反映早期DR的敏感指标。方法 对16例(32只眼)正常人进行OPs和F-ERG检测。对27例(53只眼)糖尿病病人进行眼底荧光血管造影(FFA)、OPs和F—ERG检测。结果 OPs中Os波幅、O4波幅、OPs总波幅及F-ERG中b波峰潜时较其他指标敏感，其中O4波幅和b波峰潜时为最敏感指标，但均不能反映早期DR的严重程度。结论 OPs的O4波幅和F—ERG的b波峰潜时可作为早期糖尿病视网膜病变诊断的敏感指标。     

In vivo quantification of fluorescent molecular markers in real-time: A review to evaluate the performance of five existing methods

With the advent of molecular-targeted fluorescent markers, there is a renewed interest in fluorescence quantification methods that are based on continuous wave excitation and multi-spectral image acquisition. However, little is known about their in vivo quantification performance. We reviewed the performance of five selected methods by analytically describing these and varying input parameters of irradiance, excitation geometry, collection efficiency, autofluorescence, melanin content, blood volume, blood oxygenation and tissue scattering using optical properties representing those for human skin. We identified one method that corrects for variations in all parameters. This requires image acquisition before and after marker administration, under identical geometry. Hence, it is suited for applications where the site of interest can be relocated (e.g. anaesthetized animals and dermatology). For applications where relocation is not possible, we identified a second method where the uncertainty in the fluorescence signal was ±20%. Hence, use of these methods can substantially aid in vivo fluorescence quantification compared to use of the raw fluorescence signal, as this changed by more than 3 orders of magnitude. Since these methods can be computed in real-time, they are of particular interest for applications where direct feedback is critical, as diagnostic screening or image-guided surgery.     

荧光定量PCR与ELISA法检测HBV-DNA结果分析

目的 :应用荧光定量PCR方法精确检测HBV -DNA的拷贝数 ,为临床判断病毒复制程度提供指标 ;对比分析荧光定量PCR方法和ELISA方法检测HBV -DNA的结果。方法 :用荧光定量PCR法检测 10 14例用ELISA法诊断为乙型肝炎病人的血清HBV -DNA拷贝数。结果 :10 14例病人中 ,病毒拷贝数≥ 1× 10 5的 4 99例 ,阳性率4 9.2 % ;HBeAg阳性患者的HBV -DNA拷贝数≥ 1× 10 5的阳性率显著高于HBeAg阴性患者。结论 :荧光定量PCR检测HBV病毒复制情况结果准确 ,对临床工作指导意义更大 ;HBeAg是一项重要指标。     

Fluorescence behaviour of human arterial tissue

Total fluorescence from arterial tissue is influenced by three factors: the absorption coefficient of tissue at a specific excitation wavelength, the laser excitation power and the fluorescence coefficient which is related to chemical species in tissue. These various influences were demonstrated by the following experimental results in vitro: (1) the effect of increasing power on fluorescence intensity, (2) the total fluorescence intensity in normal aorta and plaque and (3) the effect of a chromophore such as β-carotene on total fluorescence intensity. The fluorescence intensity of normal artery is an incremental function of laser excitation power, and the fluorescence emission from normal artery compared to fluorescence emission from plaque is significantly different at the same excitation power. The total fluorescence of normal artery was measured to be twice as great as that of atheromatous plaque (relative mean ratio of 2.58±0.46 compared to unity,p<0.0002 at 488 nm; relative mean ratio of 2.57±0.51 compared to unity,p<0.0009 at 514 nm). The total fluorescence emission decreases with the increase of β-carotene content in arterial tissue (R=0.97). These emission differences, when intensified by an exogenous chromophore of β-carotene, may provide an improved guidance signal for diagnosis of plaque from normal artery during laser angioplasty procedures.     

荧光分光光度法测定水产品中烷烃类、芳烃类污染物

目的：对使用荧光分光光度法测定水产品中烷烃类、芳烃类污染物的方法进行验证,并检测三类样品。方法：通过氢氧化钠-乙醇皂化处理,用正已烷萃取,用石油醚溶解,测量萃取物荧光强度值进行定量。结果：测得标准油在0.00～12.0μg/ml范围内线性关系良好,相关系数0.9999,检出限为0.2 mg/kg,平均回收率90.2%,RSD5.4%。结论：本法准确度、灵敏度高,检出限低,操作稳定性好。