MR-DWI成像预测乳腺癌腋窝淋巴转移的价值

本研究探讨磁共振弥散加权成像(MR-DWI)评估乳腺癌患者腋窝淋巴结转移的价值。回顾性选取乳腺癌患者68例(观察组),同时选取乳腺良性病变患者60例作为对照组,比较两组MR-DWI差异。观察组弥漫高信号、混杂高信号比例明显高于对照组(P<0.05);观察组病灶ADC值明显低于对照组(P<0.05);观察组Ⅲ期病灶ADC值明显低于Ⅰ~Ⅱ期(P<0.05);观察组腋窝淋巴结转移ADC值明显低于无淋巴结转移(P<0.05);弥漫高信号和其他信号组织ADC值比较差异无统计学意义(P>0.05);ADC值预测腋窝淋巴结转移的ROC曲线下面积为0.752,P<0.05。MR-DWI在乳腺癌腋窝淋巴结转移诊断应用价值较好,值得临床使用。     

Application of Deuteporfin in the Metastatic Lymph Node Mapping of Pancreatic Cancer: An in vivo Study

For most cancer patients, the presence of metastatic lymph nodes usually indicates regional recurrence and poor prognosis. Therefore, lymph node mapping is a requisite for disease staging, prognosis prediction and decision making in the treatment of cancer. Deuteporfin, a second‐generation photosensitizer, has a maximum excitation wavelength that can reach the near infrared (NIR) region (650–700 nm). We aimed to take advantage of these aspects of deuteporfin and use it as a fluorescent probe for metastatic lymph node mapping in vivo using NIR fluorescent imaging. In our study, we further investigated whether a photosensitizer could be used as a tracer for metastatic lymph node mapping of pancreatic cancer based on previous reports. Compared to normal tissues, tumor tissues including primary tumors and metastatic lymph nodes had a higher uptake ability of deuteporfin (P < 0.05). Our research confirmed this targeting property of deuteporfin using in vivo fluorescent imaging. Consistent with observations from in vivo imaging experiments, frozen sections of metastatic lymph nodes intuitively displayed significantly higher and wider distributions of deuteporfin than normal sections.     

Subsurface probing of calcifications with spatially offset Raman spectroscopy (SORS): future possibilities for the diagnosis of breast cancer

Breast calcifications are often the only mammographic features indicating the presence of a cancerous lesion. Calcium oxalate (type I) may be found in and around benign lesions, however calcium hydroxyapatite (type II) is usually found within proliferative lesions, which can include both benign and malignant pathologies. However, the composition of type II calcifications has been demonstrated to vary between benign and malignant proliferative lesions, and could be an indicator for the possible disease state. Raman spectroscopy has previously been demonstrated as a powerful tool for non-destructive analysis of tissues, utilising laser light to probe chemical composition. Raman spectroscopy is traditionally a surface technique. However, we have recently developed methods that permit its application for obtaining sample composition to clinically relevant depths of many mm. We report the first demonstration of spatially offset Raman spectroscopy (SORS) for potential in vivo breast analysis. This study evaluates the possibility of utilising SORS for measuring calcification composition through varying thicknesses of tissues (2 to 10 mm), which is about one to two orders of magnitude deeper than has been possible with conventional Raman approaches. SORS can be used to distinguish non-invasively between calcification types I and II (and carbonate substitution of phosphate in calcium hydroxyapatite) within tissue of up to 10 mm deep. This result secures the first step in taking this technique forward for clinical applications seeking to use Raman spectroscopy as an adjunct to mammography for early diagnosis of breast cancer, by utilising both soft tissue and calcification signals. Non-invasive elucidation of calcification composition, and hence type, associated with benign or malignant lesions, could eliminate the requirement for biopsy in many patients.     

High-wavenumber FT-Raman spectroscopy for in vivo and ex vivo measurements of breast cancer

The identification of normal and cancer breast tissue of rats was investigated using high-frequency (HF) FT-Raman spectroscopy with a near-infrared excitation source on in vivo and ex vivo measurements. Significant differences in the Raman intensities of prominent Raman bands of lipids and proteins structures (2,800?C3,100?cm^?1) as well as in the broad band of water (3,100?C3,550?cm^?1) were observed in mean normal and cancer tissue spectra. The multivariate statistical analysis methods of principal components analysis (PCA) and linear discriminant analysis (LDA) were performed on all high-frequency Raman spectra of normal and cancer tissues. LDA results with the leave-one-out cross-validation option yielded a discrimination accuracy of 77.2, 83.3, and 100% for in vivo transcutaneous, in vivo skin-removed, and ex vivo biopsy HF Raman spectra. Despite the lower discrimination value for the in vivo transcutaneous measurements, which could be explained by the breathing movement and skin influences, our results showed good accuracy in discriminating between normal and cancer breast tissue samples. To support this, the calculated integration areas from the receiver-operating characteristic (ROC) curve yielded 0.86, 0.94, and 1.0 for in vivo transcutaneous, in vivo skin-removed, and ex vivo biopsy measurements, respectively. The feasibility of using HF Raman spectroscopy as a clinical diagnostic tool for breast cancer detection and monitoring is due to no interfering contribution from the optical fiber in the HF Raman region, the shorter acquisition time due to a more intense signal in the HF Raman region, and the ability to distinguish between normal and cancerous tissues.     

Noninvasive Surface Detection of Papillary Thyroid Carcinoma by Fourier Transform Infrared Spectroscopy

Previously Fourier transform infrared(FTIR) spectroscopy has been applied to detecting thyroid cancer during operations and to discriminating cervical metastatic ones from non-metastatic lymph nodes. This study explored the possibility of establishing a sensitive, accurate and noninvasive screen or diagnosis by preoperative FTIR spectroscopy. 111 patients undergone a thyroid operation and 50 healthy volunteers were enrolled in the study. The FTIR spectra were obtained by two mid-infrared optical fibers with an attenuated total reflectance(ATR) probe closely contacting the subjects' skin on the thyroid nodules. The FTIR spectra obtained from normal thyroid, nodular goiter(NG) and papillary thyroid carcinoma(PTC) patients were compared. A Fisher's discriminant analysis was created based on these data. There were 41 PTC patients and 70 NG patients according to their histopathological examinations. A total of 23(of 39) parameters were statistically different among the three groups(P<0.05). The F₁₃₀₀ and F₁₀₈₀ parameters were significantly different between the three groups. In total, 9 out of 39 FTIR parameters were selected as independent factors by the Wilks' lambda stepwise discriminant analysis. The discrimination accuracy of papillary thyroid carcinoma in the three groups was 88.8%. Surface detection of PTC by FTIR spectroscopy is feasible. FTIR spectroscopy can be used for rapid and noninvasive PTC screen and auxiliary diagnosis.     

Influence of oxidative injury and monitoring of blood plasma by DSC on breast cancer patients

Damage caused by oxidative stress is involved in many types of diseases, including breast cancer. Our aim was to detect the oxidative stress parameters and blood plasma changes with differential scanning calorimetry (DSC) in breast cancer patients. The study included 40 adult breast cancer women who were grouped according to tumor diameter, regional lymph node metastases, proliferative activity, receptor status and postoperative chemotherapy. To monitor oxidative stress, malondialdehyde, oxygen free radicals (OFRs), activity of myeloperoxidase (MPO), superoxide dismutase (SOD) and catalase (CAT) were measured. Denaturation of plasma components was detected in Setaram Micro DSC-II calorimeter. The total production of OFRs, the MPO activity and lipidperoxidation were significantly increased in each breast cancer patients considering the tumor size, the metastatic lymph nodes, the proliferation activity and receptor status compared with healthy controls (p < 0.05). These pro-oxidants were slightly elevated without chemotherapy, but their values were increased significantly in chemotherapy-receiving group. The activity of SOD and CAT was significantly decreased in all groups, and in regard to the chemotherapy, they were changed significantly parallel to the severity of disease. Regarding to both the increased tumor diameter and the increased number of affected lymph nodes, DSC measurements showed a strong relationship between the maximum excess heat capacity (C_pmax) of the blood plasma and the severity of disease. The study demonstrated that oxidative stress is implicated in breast carcinoma and chemotherapy aggravates these changes which confirmed the plasma DSC measurements also.     

Distinguishing breast cancer cells using surface-enhanced Raman scattering

The detection and identification of epidermal growth factor receptor 2 (HER2)-positive breast cancer cells is crucial for the clinic therapy of breast cancer. For the aim of the detection, a novel surface-enhanced Raman scattering (SERS) probe for distinguishing breast cancers at different HER2 statuses is reported in this paper. In such a probe, anti-HER2 antibody-conjugated silver nanoparticles have been synthesized for specific targeting of HER2-positive breast cancer cells. More importantly, different from the previously reported SERS probe for targeting cancer cells, p-mercaptobenzoic acid is utilized as both the Raman reporter and the conjugation agent for attaching antibody molecules, which leads to a much simplified structure. For investigating the ability of such a probe to distinguish breast cancer cells, SKBR3 and MCF7 cells were chosen as two model systems, which are HER2-positive- and HER2-negative-expressing cells, respectively. The experimental results reveal that SKBR3 cells exhibit much stronger SERS signals than MCF7 cells, indicating that the probe could be utilized to distinguish breast cancer cells at different HER2 statuses. This kind of SERS probe holds a potential for a direct detection of living breast cancer cells with the advantages of easy fabrication, high SERS sensitivity, and biocompatibility.     

Nanoaggregate Probe for Breast Cancer Metastasis through Multispectral Optoacoustic Tomography and Aggregation-Induced NIR-I/II Fluorescence Imaging

An activatable nanoprobe for imaging breast cancer metastases through near infrared-I (NIR-I)/NIR-II fluorescence imaging and multispectral optoacoustic tomography (MSOT) imaging was designed. With a dihydroxanthene moiety serving as the electron donor, quinolinium as the electron acceptor and nitrobenzyloxydiphenylamino as the recognition element, the probe can specifically respond to nitroreductase and transform into an activated D-π-A structure with a NIR emission band extending beyond 900 nm. The activated nanoprobe exhibits NIR emission enhanced by aggregation-induced emission (AIE) and produces strong optoacoustic signal. The nanoprobe was used to detect and image metastases from the orthotopic breast tumors to lymph nodes and then to lung in two breast cancer mouse models. Moreover, the nanoprobe can monitor the treatment efficacy during chemotherapeutic course through fluorescence and MSOT imaging.     

Quantitative analysis of phosphatidylcholines and phosphatidylethanolamines in urine of patients with breast cancer by nanoflow liquid chromatography/tandem mass spectrometry

Phosphatidylcholines (PCs) and phosphatidylethanolamines (PEs) from urine of patients with breast cancer were qualitatively and quantitatively analyzed by nanoflow liquid chromatography-electrospray ionization tandem mass spectrometry (nLC-ESI-MS-MS). Urinary phospholipids (PLs) were extracted from three different categories of patients (non-cancer controls and breast cancer patients before and after surgery) by first lyophilizing only 1 mL of urine sample to enrich PLs. Next, nLC-ESI-MS-MS analysis of intact urinary phospholipids was performed, resulting in structural identification of 21 PCs and 12 PEs, followed by quantitative analysis using a multiple standard addition method. This study demonstrated that nLC-ESI-MS-MS can be powerfully utilized for the study of relative changes in the contents and concentration of urinary PCs and PEs from breast cancer patients: total concentration of PCs and PEs of patient sample increased to (144?±?9)% and (171?±?11)%, respectively, compared to control sample but they decreased significantly following surgery.     

Polymer-capped fiber-optic Raman probe for non-invasive Raman spectroscopy

Advances in fiber optic probe design are moving Raman spectroscopy into the clinic, although there remain important practical problems. While much effort has been devoted to minimizing Raman and fluorescence background from fibers, less attention has been given to the need to generate reference Raman signals that can correct for variations in tissue albedo, which is important in quantifying changes in tissue composition. To address this shortcoming, we have developed a fiber optic probe that incorporates a fluorinated ethylene-propylene copolymer (FEP) cap at the end of each excitation fiber. Transmission of laser light through the transparent cap generates a 732 cm(-1) Raman band whose intensity scales linearly with the laser power delivered to the tissue of interest. In our first design, the FEP cap functions as a waveguide with only a small insertion loss (~5%). Laser transmission through 1 mm of the polymer is sufficient to generate a usable reference Raman signal. We show the application of the probe to quantitative non-invasive Raman spectroscopy of animal tissues using rat leg phantoms as models. Ex-vivo Raman spectroscopy of excised rat tibia supports the use of the probe for spectroscopy of various tissues. These results provide proof of principle that the Raman probe can be used in multiple spectroscopic applications.     

Surface-enhanced Raman scattering detection of the breast cancer susceptibility gene BRCA1 using a silver-coated microarray platform

Surface-enhanced Raman scattering (SERS) spectroscopy was used to monitor DNA hybridization of a fragment of the BRCA1 breast cancer susceptibility gene on modified silver surfaces. Rhodamine B was covalently attached to a 5′-amino-labeled oligonucleotide sequence (23 mer) through a succinimidyl ester intermediate in methanol. The silver surfaces were prepared by depositing a discontinuous layer (9.0 nm) of silver onto glass slides, which had been etched with HF to form a microwell platform, and subsequently modified with a monolayer of mercaptoundecanoic acid. The complementary probe was covalently attached to the silver surfaces using a succinimidyl ester intermediate in acetonitrile. The silver island substrate allows a very large enhancement of the Raman signal of the DNA-Rhodamine B, and clear distinction between hybridized samples and controls on a microwell array sampling platform.     

Monitoring the absorption of organic vapors to a solid phase extraction medium applications to detection of trace volatile organic compounds by integration of solid phase absorbents with fiber optic Raman spectroscopy

By combining fiber optic Raman spectroscopy with a C-18 solid phase extraction medium, real time in situ detection of organic vapors is demonstrated. The response of the probe is fully reversible for benzene, trichloroethylene and carbon tetrachloride vapors. Because of the high degree of selectivity afforded by Raman spectroscopy, the composition of mixtures of the vapors can also be determined using the C-18 probe. The detection of ppm levels of benzene in water via headspace analysis using the C-18 probe is demonstrated.     

Raman microspectroscopy of live cells under autophagy-inducing conditions

The role of autophagy in numerous physiological responses triggered by a variety of mechanisms both in states of health and disease has raised considerable interest in this cellular process. However, the current analytical tools to study autophagy are either invasive or require genetic manipulation. Raman microspectroscopy is a potentially quantitative analytical method that has been shown to be useful for the label-free, non-destructive analysis of living biological cells and tissues. We present in this study initial efforts to study autophagy using Raman spectroscopy. The response of adherent mouse and human cancer cells to starvation conditions (glutamine deprivation and amino acid deprivation) was probed by Raman spectroscopy and compared to fluorescence microscopy results using autophagy-specific markers. We also demonstrate the capability of Raman spectroscopy to monitor the recovery dynamics of starved cells and to probe the heterogeneity in the response to starvation that can arise in cell populations. Finally, this work suggests that the 718 cm(-1) Raman line associated with phospholipids may be a useful spectral marker indicative of an autophagic response to starvation stimuli. Overall, this study establishes the utility of Raman spectroscopy to non-invasively detect biologically relevant changes in live cells exposed to conditions known to trigger autophagy.     

Nanoaggregate Probe for Breast Cancer Metastasis through Multispectral Optoacoustic Tomography and Aggregation‐Induced NIR‐I/II Fluorescence Imaging

An activatable nanoprobe for imaging breast cancer metastases through near infrared‐I (NIR‐I)/NIR‐II fluorescence imaging and multispectral optoacoustic tomography (MSOT) imaging was designed. With a dihydroxanthene moiety serving as the electron donor, quinolinium as the electron acceptor and nitrobenzyloxydiphenylamino as the recognition element, the probe can specifically respond to nitroreductase and transform into an activated D‐π‐A structure with a NIR emission band extending beyond 900 nm. The activated nanoprobe exhibits NIR emission enhanced by aggregation‐induced emission (AIE) and produces strong optoacoustic signal. The nanoprobe was used to detect and image metastases from the orthotopic breast tumors to lymph nodes and then to lung in two breast cancer mouse models. Moreover, the nanoprobe can monitor the treatment efficacy during chemotherapeutic course through fluorescence and MSOT imaging.     

A new agent for sentinel lymph node detection: preliminary results

Sentinel lymph node detection is widely used to identify lymph nodes that receive lymphatic drainage from a primary tumor. ^99mTc labeled iron oxide nanoparticles were prepared to invent a new colorful radioactive agent for sentinel lymph node detection. Iron oxide nanoparticles were produced by co-precipitation of FeCl₃ and FeCl₂ in the presence of NaOH. Then iron oxide nanoparticles were labeled with ^99mTc. ^99mTc labeled nanoparticles (7.4 MBq/0.1 mL) were intradermally injected in the distal hind limb of 16 rabbits. Dynamic and static lymphoscintigraphic images were taken for 24 h. Labeling efficiencies of ^99mTc-iron oxide nanoparticles were over 99%. Their sizes are between 50 and 60 nm. ^99mTc-iron oxide nanoparticles were accumulated in the popliteal lymph node in 11 of 16 rabbits (69%). Retention of nanoparticles in the popliteal lymph node was obvious at from 2nd through 24th hours. The radioactive lymph node was identified easily by gamma probe. The popliteal lymph node was excised and established for radioactivity and black dye. These black and radioactive nanoparticles may be potential agent successfully used for sentinel lymph node detection.     

乳腺癌术前灰阶超声与彩色多普勒超声联合应用对预后的预测价值

目的探究乳腺癌术前灰阶超声与彩色多普勒超声联合应用对预后的预测价值。方法选取温州医科大学附属第一医院2013年5月—2015年5月期间收治的乳腺癌患者83例,治疗前均行灰阶超声与彩色多普勒超声检查,分析患者超声检查结果、临床征象与术后随访2年期间患者预后的关系,从而分析术前灰阶超声与彩色多普勒超声联合应用对乳腺癌患者预后的预测价值。结果乳腺癌肿块边界清晰、边界不清晰、边界恶性晕的患者2年无瘤生存率依次降低,边界恶性晕与边界不清晰患者死亡率均显著高于边界清晰患者(P0.05);肿瘤≥2 cm患者2年无瘤生存率显著低于2 cm患者;淋巴结转移患者2年无瘤生存率显著低于无转移患者(P0.05);血流分级Ⅱ级患者2年无瘤生存率明显低于0级(P0.05),Ⅲ级患者2年无瘤生存率显著低于0级和Ⅰ级(P0.05)。结论乳腺癌患者术前联合应用灰阶超声与彩色多普勒超声检查可确定肿瘤边界、大小及淋巴转移等征象,其与患者无瘤生存率、死亡率等预后指标密切相关,可作为预测乳腺癌患者预后的重要方法。     

Raman Spectroscopy and Fluorescence Photon Migration for Breast Cancer Diagnosis and Imaging

We are developing optical methods based on near infra-red Raman spectroscopy and fluorescence photon migration for diagnosis and localization of breast cancer. We demonstrate the ability of Raman spectroscopy to classify accurately normal, benign and malignant breast tissues, an important step in developing Raman spectroscopic needle probes as a tool for improving the accuracy of needle biopsy. We also show that photon migration imaging can be used to localize accurately small fluorescent objects imbedded in a thick turbid medium with realistic optical properties, thus demonstrating the potential of this technique for optical imaging.     

Lymphatic targeting of hematoporphyrin monomethyl ether using poly (butyl-cyanoacrylate) based nanoparticle drug delivery system

The traditional treatment has inevitable drawbacks of nonspecific lymph targeting, poor therapeutic efficiency and residual metastatic for advanced cancer patients with lymph node metastases. To overcome these shortcomings, we prepare a nano-carrier drug delivery system. Photosensitizer hematoporphyrin monomethyl ether (HMME)-loaded poly (n-butylcyanoacrylate) nanoparticles (PBCA-NPs) was prepared successfully. The particle size was approximately 160 nm, the envelopment rate was 87.9%, and the drug loading rate was about 13.4%. The drug release study in vitro showed that the cumulative release rates of HMME-PBCA-NPs group was much less than free HMME group. The drug distribution in different tissues showed that the peak-reach time was 3 h in free HMME group and 6 h in nanoparticles group. All of these results confirmed the slow release characteristic of nanoparticles. In lymph node tissues, the HMME concentrations in HMME-PBCA-NPs group were much higher than those of the free HMME group at any time points we tested, in which the maximum difference concentration of HMME appeared at 6 h (1.2884?±?0.04695 vs. 0.0438?±?0.00558 µg/mg) after drug delivery. The mesenteric lymph nodes of rabbits were enlarged obviously in the NP group than in free HMME group at 6 h after drug delivery. All of these results confirmed the slow release characteristic and the lymphatic targeting characteristic of nanoparticles. In summary, we developed a lymphatic targeting nanoparticles drug delivery system successfully, which showed perfect lymph targeting and has the potential to be a new therapy strategy for advanced cancer patients with lymph node metastasis.     

Mannan-Based Nanodiagnostic Agents for Targeting Sentinel Lymph Nodes and Tumors

Early detection of metastasis is crucial for successful cancer treatment. Sentinel lymph node (SLN) biopsies are used to detect possible pathways of metastasis spread. We present a unique non-invasive diagnostic alternative to biopsy along with an intraoperative imaging tool for surgery proven on an in vivo animal tumor model. Our approach is based on mannan-based copolymers synergistically targeting: (1) SLNs and macrophage-infiltrated solid tumor areas via the high-affinity DC-SIGN (dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin) receptors and (2) tumors via the enhanced permeability and retention (EPR) effect. The polymer conjugates were modified with the imaging probes for visualization with magnetic resonance (MR) and fluorescence imaging, respectively, and with poly(2-methyl-2-oxazoline) (POX) to lower unwanted accumulation in internal organs and to slow down the biodegradation rate. We demonstrated that these polymer conjugates were successfully accumulated in tumors, SLNs and other lymph nodes. Modification with POX resulted in lower accumulation not only in internal organs, but also in lymph nodes and tumors. Importantly, we have shown that mannan-based polymer carriers are non-toxic and, when applied to an in vivo murine cancer model, and offer promising potential as the versatile imaging agents.     

Degradable intracellular self-functionalized nanoprobes for in vitro diagnosis and monitoring of cancer

Despite various attempts to accelerate nanoparticle clearance from the cell, considerable amounts of nanomaterials remain in the cell, impairing cellular activities and hence precluding reliable invitro diagnosis. Retention of nanoparticles in cellular organelles disturbs cellular equilibrium and induces long-term consequences such as change in cell phenotype, genotype, cell mobility, toxicity, and even cell death. As a result, it is necessary to produce nanoparticles that degrade promptly and are then safely expelled from the cell while remaining biocompatible and fully functional. Here, we present a metabolizable organic carbon probe functionalized for Raman and fluorescence that can be used to track and monitor metabolizable organic carbon probe breakdown in lysosomes while simultaneously performing molecular level probing for invitro cancer diagnosis. The probes were stable in medium and other fluids and degraded in cancer cells within a week. The probes demonstrated an excellent Raman signal, which allowed surface-enhanced Raman spectroscopy-based monitoring of the degradation. Additionally, the probes showed the property of fluorescence which can be attributed to the inherent fluorescence of the probe on quantum scale. Experimental evidence of self-metabolization of the probe was further demonstrated. We tested the ability of the probes for cancer diagnosis with two cancer cell lines (breast and lung) and experimentally demonstrated the ability to discriminate between breast cancer and lung cancer without any ambiguity. These findings establish a novel type of functionalized degradable nanostructure for intracellular sensing and imaging cancer cells in cellular nanomedicine.