Application of molecular probes in nuclear imaging of neuroendocrine tumors / 浙江大学学报·医学版

Neuroendocrine tumors are a type of heterogeneous tumors originating from neuroendocrine cells derived from the neural crest，which can secrete a variety of amines and peptide hormones.Based on different molecular biomarkers，histologic types and differentiation degrees，individualized nuclear imaging can provide information for the early diagnosis，clinical staging，treatment guidance，and detection of the recurrence and metastasis of neuroendocrine tumor. In this paper，we review the development and application of nuclear medicine molecular imaging probes such as glucose analogs，somatostatin analogues，amine precursors，hormone analogs and enzyme inhibitors in the diagnosis and treatment of neuroendocrine tumors.

Specificity and Targeting of Optical Molecular Probe FITC-CSPLNTRFC on Breast Cancer Cells Bcap-37 / 肿瘤防治研究

Objective To investigate the specificity and targeting abilities of FITC-CSPLNTRFC peptide FITC-BCSP1 optical molecular probe on breast cancer cells Bcap-37. Methods Probe FITC-BCSP1 and negative control probe FITC-svBCSP1 were prepared by solid phase synthesis. MTT assay was used to determine the toxicity of the two probes on breast cancer cells Bcap-37. The specificity of the binding of FITC-BCSP1 probe to Bcap-37 cells was identified by flow cytometry and fluorescent inversed microscopy. The specificity and targeting abilities of FITC-BCSP1 probe for transplantation tumor in Bcap-37 cells tumor-bearing nude mice model were tested by optical molecular imager. Results The purity of the synthesized probe was more than 98%, identified by mass spectrometry and high performance liquid chromatography. FITC-BCSP1 and FITC-svBCSP1 probes had no effect on proliferation and activity of Bcap-37 cells at the concentrations of 50-300 mol/L (IR%≤30%). FCM results showed that the percentage of FITC-BCSP1-labeled cells in Bcap-37 cells was significantly higher than that in other cells (all P < 0.001), and the percentage of FITC-BCSP1-labeled Bcap-37 cells was significantly higher than that of the control group (P < 0.001). It was observed under inverted fluorescence microscope that there were a large number of fluorescent cells in FITC-BCSP1-labeled Bcap-37 cells, with a positive rate of 100%, while the positive rate of FITC-svBCSP1 group was only 1%. In vivo assay with Bcap-37 cells tumor-bearing nude mice model showed that FITC-BCSP1 probe could be specifically enriched in the transplantation tumor tissue. Conclusion The optical molecular probe FITC-BCSP1 has good specificity and targeting abilities on breast cancer cells and can be used in the early diagnosis of breast cancer.

Preparation and application of a PDGFRβ-targeted near-infrared fluorescent probe for molecular optical imaging of lung cancer / 中国胸心血管外科临床杂志

@#Objective    To prepare platelet-derived growth factor receptor β (PDGFRβ)-targeted near-infrared molecular probe and evaluate its potential in optical molecular imaging of lung cancer. Methods    PDGFRβ-specific affibody Z-tri was recombinantly expressed in Escherichia coli (E. coli) and purified using affinity chromatography. In vitro cell-binding of Z-tri was analyzed by flow cytometry. Cellular distribution of Z-tri in tumor grafts was determined by protein-tracing. The molecular probe CF750-Z-tri was prepared by conjugating near-infrared fluorescent dye CF750 to Z-tri. The optical images of xenografts of lung cancer were obtained by using CF750-Z-tri combined with optical imaging system. Results    PDGFRβ-specific affibody Z-tri was highly expressed in E. coli and purified to homogeneity. Z-tri could bind PDGFRβ-positive cells but not PDGFRβ-negative cells cultured in vitro. In the tumor xenografts of human lung cancer, intravenously injected Z-tri was predominantly distributed on cells overexpressing PDGFRβ. The near infrared fluorescent dye CF750 was efficiently conjugated to Z-tri. Optical images with high contrast of lung cancer xenografts were produced by using the near-infrared fluorescent probe CF750-Z-tri combined with optical imaging system. Conclusion    The near-infrared fluorescent probe CF750-Z-tri can be used for optical imaging of human lung cancer, which takes great potential in optical imaging-guided surgery of lung cancer.

Research progresses of smart responsive 19F magnetic resonance nanomolecular imaging probes in diagnosis of tumors / 中国医学影像技术

Using smart responsive nanomolecular probes can realize tumor targeted imaging with rapid response, sensitive diagnosis and precise treatment under the guidance of imaging on detection of probe distributions and localization of tumors. MRI, especially 19F-MRI, has high sensitivity and low background noise. In recent years, smart responsive 19F-MR nanomolecular imaging probes had been applied for detecting changes of pH, enzyme activity, reducing microenvironment, hypoxia, light and thermo-responsive aspects. The research progresses of smart responsive 19F magnetic resonance nanomolecular imaging probes in diagnosis of tumors were reviewed in this article.

Research advances of molecular probes for early diagnosis of liver fibrosis / 中国医学影像技术

Early liver fibrosis is reversible and curable. Therefore, early diagnosis and staging of liver fibrosis are of great clinical significance. Targeting at the abnormal expression molecules in development of liver fibrosis, molecular probes are constructed as precise and non-invasive methods to integrate the information of human physiological and pathological metabolism, which is beneficial to early and specific diagnosis of fibrosis. The advances in molecular probes for early diagnosis and staging of liver fibrosis were reviewed in this article,

RGD-modified iron oxide nanoparticles for targeted molecular imaging of hepatocellular carcinoma / 实用放射学杂志

Objective To synthesize a molecular probe targeted to human hepatoma HepG2 cells with high expression of integrin αvβ3 (RGD-PEG-VSOP) and evaluate its MRI efficacy in vitro.Methods RGD-PEG-VSOP was characterized and analyzed by 1H NMR and TEM.MTT test was used to evaluate its biological safety.In vitro experiments at the cellular level,the targeting effect of RGD-PEG-VSOP to integrin was assessed,meanwhile the nontargeted nanoparticles were used as controls.Results TEM showed that the nanoparticles were spherical and uniform in size,with a relatively high r1 relaxivity of 1.37 mM-1S-1.MRI showed the signal intensity of the HepG2 cells treated with RGD-PEG--VSOP was significantly higher than that of the HepG2 cells treated with PEG-VSOP (P＜0.05).Conclusion RGD-PEG-VSOP has positive T1 contrast effect.At the cellular level,the RGD-PEG-VSOP nanoparticles have the characteristics targeted to integrin αvβ3.

MR-fluorescent dual-modality molecular probes anchored with LyP-1 for pancreatic cancer / 中国医学影像技术

Objective To construct the LyP-1 targeted MR fluorescence dual-modality molecular probe for pancreatic cancer,and to observe its features and MRI charicteristics.Methods The 50 nm MR-fluorescent dual-modality molecular probe with surface modified with cyclic nine-amino acid peptide LyP-1 (Cys-Gly-Asn-Lys-Arg-Thr-Arg-Gly Cys) was rationally designed.Whether the molecular probe could specifically recognize the pancreatic cancer cells were validated by the combination of fluorescent imaging and MR T2WI.Results The new MR-fluorescent dual-modality molecular probe anchored with LyP-1 could be used for the fluorescent imaging and MR T2WI of pancreatic cancer in mouse.And the molecular probe was demonstrated to be effective in conjugating with pancreatic cancer cells on fluorescent images and caused obvious MR signal reduction under T2 relaxometry in vitro.In vivo experiment,the molecular probe could be used for fluorescent labeling tumor tissue and detecting orthotopic pancreatic cancer in C57BL/6 mouse as MR contrast agent.Conclusion The LyP1 immobilized MR-fluorescent dual-modality molecular probe can actively target to mouse orthotopic xenograft of pancreatic cancer,which is hopeful to the application in early probing and diagnosis of pancreatic cancer by multimodal imaging.

The effect of HIFU combined with nanoscale ultrasound molecular probes with simple virus thymidine kinase gene on angiogenesis in the nude mouse tumor / 重庆医学

Objective To observe the change of the protein and gene expression of hypoxia inducing factor-1α(HIF-1a) and vascular endothelial growth factor (VEGF) in the nude mouse tumor,which has been treated by HIFU combined with nanoscale ultrasound molecular probes with HSV1-TK gene microvascular density.Methods Sixty nude mice were implanted with HepG2 Cells to establish subcutaneous transplanted tumor.Divided this mice into six groups at random after treated by HIFU:MB+ HSV-TK+ GPC3 (group A),MB + HSV-TK (group B),HSV-TK +GPC3 (group C),HSV-TK (group D),MB + GPC3 (group E),PBS (group F).They were injected into the tail vein every after 3 days.Mice in group A,B,D and E were exposed to ultrasound by 2 W/cm2,1 MHz,5 mintues and 0.2 mL ganciclovi(GCV) was intraperitoneally injected at the first 48 hours after injection.After the treatment,immunohistoche were used to detect the microvascular density(MVD),Western blot and immunohistoche was employed to test the protein change of the VEGF and HIF-1α,Q-PCR was used to test the mRNA gene transcription of VEGF and HIF-1α in the tumor tissues.Results After 14 days,the protein expression of HIF-1α and VEGF in group A was significantly lower than that in group B,C,D,E and F (P＜0.05),the MVD level in the tumor is also like this,and the difference is statistically significant.Conclusion Anoscale ultrasound molecular probes with HSV1-TK can reduce the the level of VEGF,MVD and HIF-1α in the tumor which has been treated by HIFU,so it can inhibit tumor growth and improve the therapeutic efficacy after HIFU treatment.

Bifunctional Fluorescence Molecular Probes for Detection of Aluminum and pH / 分析化学

A fluorescent molecular probe R6G-Flu was prepared by modifying fluorescein onto Rhodamine 6G.The probe could be used to recognize Al3+ specifically, and the detection limit could reach as low as 10-8 mol/L.After addition of Al3+ (10 μmol/L) to the probe, the solution showed a color change from colorless to pink, and green fluorescence was observed under the UV irradiation, which could be perceived by the naked eye.By measuring the fluorescence emission intensity of R6G-Flu at different pH, the probe could also be used to determine pH in acidic pH range (3.00-6.00) and basic pH range (8.00-10.50).The detection results of Al3+ and pH indicated that the R6G-Flu was a dual-functional fluorescent molecular probe.

Preparation of anti-EGFR-PEG-SPIO molecular probe and its targeting MRI for lung adenocarcinoma cells / 中国医学影像技术

Objective To observe the targeting function of high affinity anti-EGFR monoclonal antibody (Cetuximab)conjugated superparamagnetic iron oxide-dopamine (anti-EGFR-PEG-SPIO) lung cancer cells via epidermal growth factor receptor (EGFR),as well as the feasibility for surveillance of tumor targeting with MRI.Methods Nanoparticles (NPs)of anti-EGFR-PEG-SPIO and PEG-SPIO were prepared,and the morphology of nanoparticles was observed with transmission electron microscope (TEM).The hydrodynamic diameter and R2 values of nanoparticles before and after conjugation with anti-EGFR were performed with dynamic light scattering (DLS) and MRI.MRI was performed in incubation with anti-EGFR-PEG-SPIO and PEG-SPIO after 2 h in vitro.The cellular uptake of anti-EGFR-PEG-SPIO and PEG-SPIO was further evaluated using Prussian blue staining and TEM.Results Anti-EGFR-PEG-SPIO and PEG-SPIO showed signal intensity of H460 cells on T2WI,decreased significantly compared with PEG-SPIO.The rate of signal intensity change was -58.2％,-82.7％,-94.4％ and-98.3％,respectively,at iron concentrations of (0,10,20,40,80 μg/ml) of antiEGFR-PEG-SPIO.Prussian blue staining and TEM showed that a lot of intracellular irons of anti-EGFR-PEG-SPIO were observed in H460 cells,but few of PEG-SPIO.Conclusion The effect of active targeting via anti-EGFR in EGFR overexpressed cells can be achieved with anti-EGFR-PEG-SPIO in H460 cells in vitro,and the targeting delivery process could be monitored with 3.0T MRI.

MR-fluorescent dual-modality molecular probes anchored with LyP-1 for pancreatic cancer / 中国医学影像技术

Objective To construct the LyP-1 targeted MR fluorescence dual-modality molecular probe for pancreatic cancer,and to observe its features and MRI charicteristics.Methods The 50 nm MR-fluorescent dual-modality molecular probe with surface modified with cyclic nine-amino acid peptide LyP-1 (Cys-Gly-Asn-Lys-Arg-Thr-Arg-Gly Cys) was rationally designed.Whether the molecular probe could specifically recognize the pancreatic cancer cells were validated by the combination of fluorescent imaging and MR T2WI.Results The new MR-fluorescent dual-modality molecular probe anchored with LyP-1 could be used for the fluorescent imaging and MR T2WI of pancreatic cancer in mouse.And the molecular probe was demonstrated to be effective in conjugating with pancreatic cancer cells on fluorescent images and caused obvious MR signal reduction under T2 relaxometry in vitro.In vivo experiment,the molecular probe could be used for fluorescent labeling tumor tissue and detecting orthotopic pancreatic cancer in C57BL/6 mouse as MR contrast agent.Conclusion The LyP1 immobilized MR-fluorescent dual-modality molecular probe can actively target to mouse orthotopic xenograft of pancreatic cancer,which is hopeful to the application in early probing and diagnosis of pancreatic cancer by multimodal imaging.

Use and evaluation of a line probe assay in patients with tuberculosis in Peru: 2011 and 2013 / Utilización y evaluación de un ensayo con sondas en línea en los pacientes con diagnóstico de tuberculosis en el Perú del 2011 al 2013

ABSTRACT Objective To determine the use and performance of a line probe assay (LPA) compared with conventional culture and drug sensitivity testing (CDST) in patients registered with tuberculosis (TB) under routine program conditions in Peru in 2011–2013. Methods This was a descriptive, operational research, cross-sectional study of sputum specimens from patients with smear-positive pulmonary TB and mycobacterial cultures from patients with smear-negative or positive TB. Drug resistance to rifampicin and/or isoniazid detected by LPA was compared to CDST. Sensitivity, specificity, and predictive values were calculated and reliability for detecting drug resistance was assessed through kappa coefficient, with values 0.61–0.80 showing substantial correlation, and 0.81 or above showing almost-perfect correlation. Results In 2011–2013, there were 16 169 LPA tests performed, with the proportion of TB patients receiving the test increasing from 3.2% to 30.2%. In all, 2 905 LPA test results were compared to CDST. For LPA in sputum specimens, sensitivity for rifampicin was 92%; isoniazid, 94%; and MDR-TB, 88%; while specificity for rifampicin was 92%; isoniazid, 92%; and MDR-TB, 95%. For LPA in mycobacterial cultures, sensitivity for rifampicin was 95%; isoniazid, 96%; and MDR-TB, 90%; while specificity for rifampicin was 85%; isoniazid, 91%; and MDR-TB, 94%. Kappa coefficients were at 0.81 or above for all comparisons of LPA with CDST using sputum specimens and cultures, except for isoniazid in cultures, which was at 0.79. Conclusions This study suggests that LPA is a reliable and rapid screening test for drug-resistant TB and should be considered suitable for routine use and scale up in Peru.

RESUMEN Objetivo Definir la utilización de un ensayo con sondas en línea y evaluar su desempeño, en comparación con el método convencional de cultivo y antibiograma, en los pacientes registrados con tuberculosis en condiciones programáticas en el Perú del 2011 al 2013. Métodos Investigación operativa descriptiva con un estudio transversal de las muestras de esputo de los pacientes con diagnóstico de tuberculosis pulmonar y baciloscopia positiva y de los cultivos de micobacterias de los pacientes con tuberculosis y baciloscopia positiva o negativa. La farmacorresistencia a la rifampicina, la isoniacida o a ambas, detectada mediante el ensayo con sondas en línea, se comparó con los resultados obtenidos por el método de cultivo y antibiograma. Se calculó la sensibilidad, la especificidad y los valores predictivos del ensayo con sondas en línea y se evaluó su fiabilidad en la detección de la farmacorresistencia mediante el coeficiente k, cuyos valores de 0,61 a 0,80 correspondían a una fuerte correlación y los valores de 0,81 o superiores reflejaban una correlación casi perfecta. Resultados Del 2011 al 2013 se practicaron 16 169 ensayos con sondas en línea, y la proporción de pacientes con diagnóstico de tuberculosis en quienes se practicaba aumentó de 3,2% a 30,2%. En total, se compararon 2 905 resultados del ensayo molecular con el método convencional. En las muestras de esputo, el ensayo molecular ofreció una sensibilidad de 92% para la resistencia a la rifampicina, 94% a la isoniacida y 88% para la tuberculosis multirresistente; su especificidad fue 92% con respecto a la rifampicina, 92% a la isoniacida y 95% a la tuberculosis multirresistente. En los cultivos de micobacterias, el ensayo con sondas en línea mostró una sensibilidad de 95% para la resistencia a la rifampicina, 96% a la isoniacida y 90% para la tuberculosis multirresistente; la especificidad fue 85% para la rifampicina, 91% para la isoniacida y 94% para la tuberculosis multirresistente. El coeficiente k fue 0,81 o superior en todas las comparaciones del ensayo molecular con el método tradicional cuando se usaron muestras de esputo y cultivo, excepto con la isoniacida en cultivo, cuyo coeficiente fue 0,79. Conclusiones Los resultados del presente estudio indican que el ensayo con sondas en línea constituye una prueba de detección fiable y rápida para la tuberculosis multirresistente, y se debe considerar apropiada su utilización en la práctica de rutina y la ampliación de su empleo en el Perú.

Significance of modified gold nanocages in photothermal therapy and radiosensitivity enhancement / 中华放射肿瘤学杂志

Objective To investigate the significance of modified gold nanocages ( GNCs ) in photothermal therapy and radiosensitivity enhancement for murine mammary carcinoma 4T1 cells.Methods A molecule probe, CD44-PEG-GNCs, was constructed on the basis of GNCs.Inductively coupled plasma mass spectrometry ( ICP-MS) and transmission electron microscopy ( TEM) were used to analyze Au uptake by 4T1 cells.The cell counting kit-8( CCK-8) assay was used to determine the impact of the probe on the viability of 4T1 cells.The CCK-8 assay and the Hoechst/PI double staining were used to evaluate the photothermal killing effects of near infrared laser (NIR).The colony forming assay and the Hoechst/PI double staining were used to measure the viability of 4T1 cells after irradiation with 6 MV of X-ray.Results ICP-MS and TEM showed that 4T1 cells had an uptake of a large number of GNCs, in which the number of CD44-PEG-GNCs taken up by 4T1 was 3-4 times larger than that of PEG-GNCs.The cell toxicity assay revealed that in a certain range of concentration, GNCs showed little effect on the viability of 4T1 cells, which was reduced to 81.2%in 48 hours.The CCK-8 assay, the Hoechst/PI double staining, and the colony forming assay showed that after NIR irradiation and radiotherapy, large-scale apoptosis was induced in CD44-PEG-GNCs-contained tumor cells by photothermal therapy combined with radiotherapy, which had substantially better outcomes than other groups.Conclusions The GNC-based molecular probe actively targets 4T1 cells with little cytotoxicity, indicating that it enhances the sensitivity of tumor towards photothermal therapy and radiotherapy.

Research on biological activities and MRI of differentiations of neural-like cells induced by superparamagnetic iron oxide and green fluorescent protein double-labeled bone marrow-derived mesenchymal stem cells in vitro / 中华放射学杂志

Objective To explore the biological activities and the MR imaging signal intensities (SIs) characteristics of differentiations of neural-like cells induced by superparamagnetic iron oxide (SPIO) and green fluorescent protein (GFP) double-labeled bone marrow-derived mesenchymal stem cells (BMSCs) in vitro. Methods GFP-BMSCs were labeled with different concentrations of SPIO in vitro (the concentration of the A, B, C and D group was 25, 50, 75 and 100 ug/ml, respectively;the E group without labels of SPIO served as the control group). The Prussian blue stainings were used to detect the labeling rates of SPIO. The iron contents of cells were measured by atomic absorption spectrometer. The CCK8 experiments were used to detect the cell proliferation rates. The cell cycles were detect by PCR. Each of the A-E groups had a test tube with 1 × 108 cells. All test tubes underwent T2* weighted imaging (T2*WI) and susceptibility weighted imaging (SWI) in a MR imaging scanner. The optimal group was defined by comparing the measurements of SIs between T2*WI and SWI. The optimal group and the E group together induced the differentiations of osteogenesis and neural-like cells. The stainings of alizarin red, osteocalcin and Nissl, NeuN, and NF-200 were performed at 72 hours. Real-time quantitative PCR was used to detect the expression levels of RNA in tub3, nestin, NSE, MAP-2 and Syt1. The positive staining rates and the expression levels of RNA were compared between the two groups. Finally, SWI was used to analyse the changes of SIs. One-way analysis of variance (ANOVA) was used to the multi-group comparison. Using least significant difference (LSD) test to analyse the comparisons between the multi-groups. Results The labeling rates of the A-D groups were 100%. The iron contents of cells in the A-E groups were (14.36 ± 7.61), (21.73±3.42), (30.54±8.73), (33.65±9.62), and (2.31±0.32) pg/cell, respectively. The iron contents of cells in the A-D groups were significantly higher than those in the E group ( F=3.852, P=0.003). There was no significant difference between the C and D groups (P=0.267). In all groups, the D group had the lowest OD value in the CCK-8 experiments (3.18 ± 0.46). In the A-E groups, the changes of SIs in SWI were significantly lower than those in T*2 WI. There was no significant difference in SIs of SWI between the C group (145.89±14.31) and the D group (127.37±12.21). Except the comparison between the group C and D, the comparisons between all the groups had significant differences (P0.05). The expression levels of tub3, nestin and NSE were significantly higher before than after induced differentiations (P<0.01). SIs of SWI had no significant difference between before and after induced differentiations in the C group (t=1.26, P=0.236). Conclusions SPIO and GFP double-labeled BMSCs can induce neural-like cells without influencing biologic activities. MR SIs are decreased by the increase of SPIO concentrations in cells. SWI was the most sensitive sequence. The SIs of SWI has no differnce between before and after induced differentiations.

Progress of Multimodal Molecular Imaging Technology in Diagnosis of Tumor / 分析化学

The multumodal molecular umagung technology untegrates the advantages of varuant umagung methods whuch can provude a more comprehensuve and accurate unformatuon un cancer duagnosus, and realuze tumely personaluzed duagnosus of tumor at molecular and cellular level, quantutatuvely dynamuc monutorung of tumor, etc. Thus revuew untroduces the basuc concepts of multumodal molecular umagung, umplementatuon methods and recent research progress of the applucatuons un tumor duagnosus. The development trend of multumodal molecular umagung un tumor duagnosus us also prospected.

Fabrication of molecular probe targeting NRP-1 and magnetic resonance molecular imaging of gliomas in vitro / 中华行为医学与脑科学杂志

Objective To synthetize a novel MR molecular imaging probe named USPIO?PEG?tLyP?1,and to evaluate its value in detecting U87 cells by MR imaging. Methods USPIO?PEG?tLyP?1 was synthetized by conjugating USPIO?PEG with tLyP?1. Neuropilin?1 expression levels of glioma cell lines were detected by Western blot. The cytotoxicity of USPIO?PEG and USPIO?PEG?tLyP?1 were assessed by MTT colorimetric assay. The uptake efficiency of USPIO?PEG?tLyP?1 was measured by Prussian blue staining, transmission electron microscope and MR imaging in vitro. Results The novel MR molecular imaging probe was synthetized with an average diameter of 43.84 nm. U87 glioma cell line was screened as test subject for the highly expression of NRP?1(P<0.05). USPIO?PEG?tLyP?1 group showed much more intracellular blue particles than USPIO?PEG group after Prussian blue staining. After incubation,USPIO?PEG?tLyP?1 mainly existed in lysosme,endoplasmic reticulum and mitochondria. In vitro MRI showed that USPIO?PEG?tLyP?1 significantly enhanced the negative contrast effect compared with USPIO?PEG(P<0.01). Conclusion The decoration of tLyP?1 enhanced targeting ability of USPIO?PEG to glioma cells and MR molecular imaging can be a promising method for early diagnosis of gliomas.

Experimental study on the characters of the FITC-CSNRDARRC molecular probe and its targeted ability for orthotopic transplanted tumor of bladder cancer in vivo / 中华泌尿外科杂志

Objective To investigate the characters and targeted ability of FITC-CSNRDARRC molecular probe in labeling orthotopic transplantation tumor of bladder cancer in vivo.Methods From July 2013 to June 2014,the stability and characters of FITC-CSNRDARRC molecular probe were detected by spectrophotometer and molecular imaging and the optimum concentration and imaging time window were determined.30 BALB-C nude mice were randomly divided into experimental group (n =20) and control group(n =10).In control group,5 of them (group A) were ligated bilateral ureter,others(group B) were not.We established orthotropic transplanted bladder tumor (BIU-87) model by operation.And 0.2 ml probes (220 μmol/L) was then injected intravenously in all mice after 2 weeks.We obtained images and analyzed average gray value of the heart,lung,liver,spleen,bilateral kidney and orthotropic transplantation bladder tumor by using optical probe molecule fluorescence imaging system after 30 min,1 h,2 h,4 h and 12 h,respectively.Results After injected the FITC-CSNRDARRC molecular probes intravenously at 220 μmol/L,the fluorescence signal of tumor tissue strengthened gradually.The optimal imaging time window was 4 hours after injection.The illumination and temperature had little effect on the fluorescence signal.With the time passing after injection,the intensity of florescence signal progressively increasing,which reached the peak at4 h.The average gray value of tumor tissue at 1 h,2 h,3 h,4 h,5 h,6 h,8 h,12 h were 74.22,76.2,80.11,89.38,83.29,85.1,81.22,83.01,respectively.The fluorescence signal of normal tissue weakened gradually with the passage of time.Only liver and gall bladder could notice the fluorescence signal 4 hours after injection in group A.However,the relatively strong fluorescence signal could be found in liver and gall bladder in group B.Conclusions The characters of fluorescence probe are affected by its concentration.Its optimal concentration of labeling tumor is 220 μmol/L.The optical imaging time window was about 4 h after intravenous injection.The FITC-CSNRDARRC molecular probe can specifically bound to orthotopic transplanted tumor of bladder cancer in vivo.

Preclinical study of bladder cancer diagnosis using tumor homing peptide fluorescent molecular probe / 中华泌尿外科杂志

Objective To evaluate the features of optical molecular imaging of bladder tumor cells labeled by tumor homing peptide fluorescent molecular probe, and to explore the theoretical foundation of optical molecular imaging for bladder cancer diagnosis.Methods After prepared the FITC-CSNRDARRC fluorescent molecular probe, laser scanning confocal microscope, immuno fluorescence and multispectral fluorescence in vivo optical molecular imaging system have been used to evaluate the binding sites, the affecting factors of binding rates, the specificity and the targets.BIU-87 bladder tumor cell line, BIU-87 bladder tumor cell line, 68 cases of paraffin bladder tumor tissue samples, 16 cases of paraffin glandular bladder inflammatory samples, 43 cases of paraffin renal clear cell carcinoma samples, 68 cases of paraffin gastric adenocarcinoma samples, 29 cases of urine exfoliated cells suspected bladder cancer and BIU-87 bladder cancer nude xenograft have been used in this study.Results The binding site of FITC-CSNRDARRC fluorescent molecular probe were at the nucleus of labeled bladder tumor cells.The binding rates were correlated linearly with the dose of probe and the grade of pathology.The in vitro and in vivo studies demonstrate that the probe has a binding specificity with bladder tumor. When the FITC-CSNRDARRC fluorescent molecular probe labeled tumor cells, bright green spots were observed under laser scanning confocal microscope.The bright green spots were more apparent after stained by DAPI again.The tissue samples and tumor cells in the urine can be successful labeled and identified by fluorescence microscope.Optical molecular imaging of in vivo xenograft tumor tissues showed fluorescent spots under EMCCD.Conclusions The labeled loci of single cell by FITC-CSNRDARRC probe have been identified. The spatial resolution of optical molecular image is related to sensitivity of CCD, and the optical molecular imaging cannot be imaged by the conventional endoscope camera.

Use of amplified Mycobacterium tuberculosis direct test in respiratory samples from HIV-infected patients in Brazil / Utilização do amplified Mycobacterium tuberculosis direct test em amostras respiratórias de pacientes HIV positivos no Brasil

OBJECTIVE: To compare the accuracy of the amplified Mycobacterium tuberculosis direct (AMTD) test with reference methods for the laboratory diagnosis of tuberculosis in HIV-infected patients. METHODS: This was a study of diagnostic accuracy comparing AMTD test results with those obtained by culture on Löwenstein-Jensen (LJ) medium and by the BACTEC Mycobacteria Growth Indicator Tube 960 (BACTEC MGIT 960) system in respiratory samples analyzed at the Bioassay and Bacteriology Laboratory of the Oswaldo Cruz Foundation Evandro Chagas Clinical Research Institute in the city of Rio de Janeiro, Brazil. RESULTS: We analyzed respiratory samples collected from 118 patients, of whom 88 (74.4%) were male. The mean age was 36.6 ± 10.6 years. Using the AMTD test, the BACTEC MGIT 960 system, and LJ culture, we identified M. tuberculosis complex in 31.0%, 29.7%, and 27.1% of the samples, respectively. In comparison with LJ culture, the AMTD test had a sensitivity, specificity, positive predictive value, and negative predictive value of 87.5%, 89.4%, 75.7%, and 95.0%, respectively, for LJ culture, whereas, in comparison with the BACTEC MGIT 960 system, it showed values of 88.6%, 92.4%, 83.8%, and 94.8%, respectively. CONCLUSIONS: The AMTD test showed good sensitivity and specificity in the population studied, enabling the laboratory detection of M. tuberculosis complex in paucibacillary respiratory specimens. .

OBJETIVO: Comparar a acurácia do teste amplified Mycobacterium tuberculosis direct (AMTD) com métodos de referência para o diagnóstico laboratorial de tuberculose em pacientes HIV positivos. MÉTODOS: Estudo de acurácia diagnóstica comparando os resultados do teste AMTD com os de cultura em Löwenstein-Jensen (LJ) e de BACTEC Mycobacteria Growth Indicator Tube 960 (sistema BACTEC MGIT 960) em amostras respiratórias analisadas no Laboratório de Bacteriologia e Bioensaios do Instituto de Pesquisa Clínica Evandro Chagas da Fundação Oswaldo Cruz, no Rio de Janeiro (RJ). RESULTADOS: Foram analisadas amostras respiratórias de 118 pacientes, dos quais 88 (74,4%) eram do sexo masculino. A média de idade foi de 36,6 ± 10,6 anos. O complexo M. tuberculosis foi identificado em 31,0%, 29,7% e 27,1% das amostras através do teste AMTD, sistema BACTEC MGIT 960 e LJ, respectivamente. Na comparação com a cultura em LJ, o teste AMTD apresentou sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo de 87,5%, 89,4%, 75,7% e 95,0%, respectivamente, enquanto na comparação com o sistema BACTEC MGIT 960, os valores foram de 88,6%, 92,4%, 83,8% e 94,8%, respectivamente. CONCLUSÕES: O teste AMTD mostrou boa sensibilidade e especificidade na população estudada, possibilitando a detecção laboratorial do complexo M. tuberculosis em espécimes respiratórios paucibacilares. .

Molecular inversion probe-based single base mutations of hepatitis B virus resistance gene analysis / 中华检验医学杂志

Objective To establish a molecular inversion probe ( MIP) method for detection of single base drug-resistance mutation in Hepatitis B virus ( HBV) gene.Methods The HBV wild type and YVDD mutant strain were isolated by Daping Hospital of the Third Military Medical University.The MIP was designed and applied to detect the HBV drug-resistance YVDD mutation in one case of wild type and one case of YVDD mutant HBV strain isolated previously.The results of MIP method were compared with that of sequencing to evaluate the detection accuracy.Results Thermal cycling single-base extension and connection reaction performed by Taq DNA Ligase and Ampligase DNA Ligase could ensure the specificity of the detection.The optimum probe concentration of MIP was 1 nmol/L.Through detection of the target gene with different DNA concentrations , the detection sensitivity of MIP was determined as 1 nmol/L.The results of MIP were consistent with that of sequencing method in detection of the clinical samples.Conclusion MIP is successfully used to detect single-base drug-resistance mutation in HBV gene.