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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.
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Objective:To investigate the property and the combination effect of the peptide specifically binding to human medullary breast cancer Bcap-37 cells by using phage display in vivo and to provide molecular targeting probe for early diagnosis of breast cancer.Methods:The human medullary breast carcinoma Bcap-37 cells tumor-bearing nude mice model was prepared and three rounds in vivo were performed by using Ph.D.-C7CTM phage display peptide library. The distribution of screened phages in tumors and normal tissues was detected by using immunohistochemistry. The affinity of monoclonal phage to Bcap-37 cells was identified by using enzyme linked immunosorbent assay (ELISA). The positive monoclonal phage DNA was taken and sequenced, and the sequence with high repetition rate was selected to synthesize peptide by using chemical methods. Optical molecular probe was prepared and fluorescence molecular imaging was used to test its specificity and targeting ability for breast transplantation tumor of tumor-bearing nude mice in vivo.Results:The recovery rate of phage in the third round screening in vivo was 107.2 times than that in the first round. Immunohistochemical results showed that the phages binding to the tumor tissues were increased successively with the increasing screening rounds in vivo. The number of phages binding to tumor tissues was more than that binding to normal tissues (lung, skeletal muscle, liver and kidney). The absorbancy ( A) value of section scanning image in the tumor tissues was higher than that in the normal tissues, and the difference was statistically significant different ( P < 0.05). ELISA results showed that 22 phages (affinity≥2) among the 50 randomly selected monoclonal phage were positive. After DNA sequencing analysis of the positive monoclonal phage, 4 repeat amino acid sequences were obtained. The fluorescein isothiocyanate-labelled (FITC)-CSPLNTRFC with the highest repetition rate was synthesized for FITC-CSPLNTRFC peptide. Bcap-37 cells tumor-bearing nude mice model assay in vivo showed that FITC-CSPLNTRFC peptide could significantly enriched in the breast xenograft tissues. Conclusion:CSPLNTRFC peptide specifically binding to human medullary breast cancer Bcap-37 cells can be screened out successfully by using phage display in vivo, which will be helpful to vitro studies on the early diagnosis of breast cancer.
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Parkinson disease (PD) is the second-most common neurodegenerative disorder. Its main pathological mechanism is the selective degeneration and deletion of dopaminergic neurons in the dense part of the substantia nigra and the damage of dopaminergic neurons caused by the abnormal deposition of a Lewy body, leading to a decreased dopamine level. Positron emission computed tomography (PET)/single photon emission computed tomography (SPECT) is a molecular imaging technology that can directly or indirectly reflect changes in molecular levels by using a specific tracer. With the research and development on the tracers of related enzymes for labeling dopamine transporter and dopamine receptor and for being involved in dopamine formation, this imaging technology has been applied to all aspects of PD research. It not only contributes to clinical work but also provides an important theoretical basis for exploring the pathological mechanism of PD at a molecular level. Therefore, this review discusses the application value of PET/SPECT in PD in terms of early diagnosis, disease severity evaluation, clinical manifestations, differential diagnosis, and pathological mechanism.
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BACKGROUND@#Non-small cell lung cancer (NSCLC) is the most common histological type of lung cancer, and one of the malignant tumor with the highest mortality. As the main part of the optical molecular imaging probe, peptide can realize the early screening and diagnosis of tumor and improve the survival rate of patients. The aim of this study was to screen the small-molecule peptide that highly binds to NSCLC NCI-H1299 cells using in vivo phage display technology and to identify their binding specificity by in vitro experiment.@*METHODS@#To prepare a tumor-bearing nude mouse model of NCI-H1299 cells, after 3 rounds of in vivo screening with Ph.D.-C7CTM Peptide Library, phage clones were randomly picked, using immunohistochemistry and enzyme-linked immunosorbent assay (ELISA) to identify the affinity of phage clones to NCI-H1299 cells. The positive monoclonal phages DNA was extracted and sequenced to obtain the amino acid sequence of the peptides. The peptides with the highest repetition rate was chemically synthesized and labeled with fluorescein (FITC) to prepare optical molecular probe. We preliminary identified the specificity of the probe binding to lung cancer cells by in vitro experiment.@*RESULTS@#After three rounds of in vivo screening, the phages enrichment rate was 341.3 times compared with the first round. Immunohistochemical staining showed that with the increase of screening times, the phages binding to tumor tissues continued to increase, and the binding amount was significantly higher than normal tissues; ELISA results showed that 20 clones among the 30 randomly selected phage clones were positive. After sequencing, the peptide with the highest repetition rate was synthesized and named NSP1; Methyl thiazolyl tetrazolium assay (MTT) and would healing assay showed that NSP1 will not affect cell proliferation and migration. Flow cytometry and immunofluorescence showed specific binding of NSP1 to NCI-H1299 cells.@*CONCLUSIONS@#We successfully obtained the peptide NSP1 that specifically binds to lung cancer NCI-H1299 cells by in vivo phage display, which provide a theoretical basis for NSCLC early diagnosis and targeted therapy.
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Purpose To evaluate the clinical guiding value of 99Tcm-methoxyisobutylisonitrile gated myocardial perfusion imaging (99Tcm-MIBI G-MPI) in diagnosing the coronary microangiopathy in patients with diabetic mellitus.Materials and Methods Sixty patients with clinical confirmed coronary microvascular angina were selected and assigned into two groups,with 26 patients (with diabetes mellitus) in group A and 34 patients in group B (without diabetes mellitus).The region,extent and range of coronary microangiopathy was detected by 99Tcm-MIBI G-MPI,and the difference of myocardial ischemia between the two groups was statistically analyzed.Results The ratio of abnormal myocardial perfusion in group A was significantly higher than that in group B (96.15％ vs.73.53％,x2=5.43,P＜0.05);the total number of abnormal coronary branches in group A was significantly larger than that in group B (82.67％ vs.62.67％,P＜0.05);the total number abnormal perfusion segments in group A was significantly larger than that in group B (P＜0.05),and the abnormal perfusion segments number of anterior wall,septum,apical region and inferior wall in group A was significantly larger than that ingroup B (P＜0.05);the rate of abnormal perfusion at single coronary supplying region in group A was significantly lower than that in group B (P＜0.05).Conclusion 99Tcm-MIBI G-MPI is of great clinical significance for diagnosing coronary microangiopathy and evaluating myocardial ischemia in patients with diabetes mellitus.
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Due to its exquisite sensitivity, excellent molecular specificity and reduced photobleaching, surface-enhanced Raman spectroscopy has wide potential applications in the fields of food and drug analysis, environmental monitoring and biological detection. However, SERS is only a qualitative or semi-quantitative detection technique at the present stage, and has not yet been widely recognized as a routine quantitative technique. This review discusses the pros and cons of the quantitative surface-enhanced Raman spectroscopic techniques developed so far in the literature, and looks forward to the future trend in this field.
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Chromosomal integration enables stable phenotype and therefore has become an important strategy for breeding of industrial Saccharomyces cerevisiae strains. pAUR135 is a plasmid that enables recycling use of antibiotic selection marker, and once attached with designated homologous sequences, integration vector for stable expression can be constructed. Development of S. cerevisiae strains by metabolic engineering normally demands overexpression of multiple genes, and employing pAUR135 plasmid, it is possible to construct S. cerevisiae strains by combinational integration of multiple genes in multiple sites, which results in different ratios of expressions of these genes. Xylose utilization pathway was taken as an example, with three pAUR135-based plasmids carrying three xylose assimilation genes constructed in this study. The three genes were sequentially integrated on the chromosome of S. cerevisiae by combinational integration. Xylose utilization rate was improved 24.4%-35.5% in the combinational integration strain comparing with that of the control strain with all the three genes integrated in one location. Strain improvement achieved by combinational integration is a novel method to manipulate multiple genes for genetic engineering of S. cerevisiae, and the recombinant strains are free of foreign sequences and selection markers. In addition, stable phenotype can be maintained, which is important for breeding of industrial strains. Therefore, combinational integration employing pAUR135 is a novel method for metabolic engineering of industrial S. cerevisiae strains.
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Engenharia Genética , Métodos , Vetores Genéticos , Engenharia Metabólica , Plasmídeos , Genética , Saccharomyces cerevisiae , Genética , Xilose , MetabolismoRESUMO
Objective To discuss the effect of community nursing intervention on medication compliance and control rates of blood pressure in patients with essential hypertension.Methods Community nursing intervention was conducted to 73 community patients with essential hypertension. The patients' medication compliance was compared before and after intervention and their control of blood pressure was also observed. Results The difference was statistically significant in the comparison of patients' knowledge of antihypertension and the proper use of antihypertensive drugs before and after intervention. Blood pressure control rate was 80.82%.Conclusions Community nursing intervention can improve patients' medication compliance and effectively control the blood pressure of the patients.