Exosome, a Rising Biomarkers in Liquid Biopsy: Advances of Label-Free and Label Strategy for Diagnosis of Cancer.

Cancer is commonly considered as one of the most severe diseases, posing a significant threat to human health and society due to various serious challenges. These challenges include difficulties in accurate diagnosis and a high propensity to form metastasis. Tissue biopsy remains the gold standard for diagnosing and subtyping cancer. However, concerns arise from its invasive nature and the potential risk of metastasis during these complex diagnostic procedures. Meanwhile, liquid biopsy has recently witnessed the rapid advancements with the emergence of three prominent detection biomarkers: circulating tumor cells (CTCs), circulating tumor DNA (ctDNA), and exosomes. Whereas, the very low abundance of CTCs combined with the instability of ctDNA intensify the challenges and decrease the accuracy of these two biomarkers for cancer diagnosis. While exosomes have gained widespread recognition as a promising biomarker in liquid biopsy due to their relatively low-invasive detection method, excellent biostability, rich resources, high abundance, and ability to provide valuable information about cancer. Therefore, it is crucial to systematically summarize recent advancements mainly in exosome-based detection methods for early cancer diagnosis. Specifically, this review will primarily focus on label-based and label-free strategies for detecting cancer using exosomes. We anticipate that this comprehensive analysis will enhance readers' understanding of the significance and value of exosomes in the fields of cancer diagnosis and therapy.

A Stabilized Li-Metal Anode with a Ti-Based Metal-Organic Framework Electronic Shield.

The insufficient cyclic efficiency and poor safety have prohibited the commercial applications of the lithium-metal anode because of its uncontrolled dendrite growth at the surface. A mechanically stable and highly ionic conductive solid electrolyte interphase (SEI) holds great promise to address the issues. Herein, a viable surface engineering approach is proposed for stabilizing the Li anode via a scalable artificial method. The surface of Li metal is functionalized by constructing a mechanically tough and electron-insulating metal-organic framework (MOF) of the MIL-125(Ti) layer. In-situ optical microscopy reveals its crucial role in inhibiting dendritic Li growth. Because of the intrinsic insulativity and highly ordered micropores of MIL-125(Ti), the Li+ ions acquire electrons under the coating layer, resulting in a uniform and dense Li deposition behavior. The symmetric cell of the MOF-modified Li electrode delivers a long life span of 2000 h with an overpotential of less than 20 mV at 0.5 mA cm-2. When paired with the same MOF-derived sulfur cathode, decent cycling retention is available as well. This work demonstrates a feasible strategy for the development of a stable Li-metal anode with alleviative dendritic growth.

Hydralazine Promotes Central Nervous System Recovery after Spinal Cord Injury by Suppressing Oxidative Stress and Inflammation through Macrophage Regulation.

OBJECTIVE: This study aims to investigate the effects of hydralazine on inflammation induced by spinal cord injury (SCI) in the central nervous system (CNS) and its mechanism in promoting the structural and functional recovery of the injured CNS. METHODS: A compressive SCI mouse model was utilized for this investigation. Immunofluorescence and quantitative real-time polymerase chain reaction were employed to examine the levels of acrolein, acrolein-induced inflammation-related factors, and macrophages at the injury site and within the CNS. Western blotting was used to evaluate the activity of the phosphoinositide 3-kinase (PI3K)/AKT pathway to study macrophage regulation. The neuropathic pain and motor function recovery were evaluated by glutamic acid decarboxylase 65/67 (GAD65/67), vesicular glutamate transporter 1 (VGLUT1), paw withdrawal response, and Basso Mouse Scale score. Nissl staining and Luxol Fast Blue (LFB) staining were performed to investigate the structural recovery of the injured CNS. RESULTS: Hydralazine downregulated the levels of acrolein, IL-1ß, and TNF-α in the spinal cord. The downregulation of acrolein induced by hydralazine promoted the activation of the PI3K/AKT pathway, leading to M2 macrophage polarization, which protected neurons against SCI-induced inflammation. Additionally, hydralazine promoted the structural recovery of the injured spinal cord area. Mitigating inflammation and oxidative stress by hydralazine in the animal model alleviated neuropathic pain and altered neurotransmitter expression. Furthermore, hydralazine facilitated motor function recovery following SCI. Nissl staining and LFB staining indicated that hydralazine promoted the structural recovery of the injured CNS. CONCLUSION: Hydralazine, an acrolein scavenger, significantly mitigated SCI-induced inflammation and oxidative stress in vivo, modulated macrophage activation, and consequently promoted the structural and functional recovery of the injured CNS.

An attention transfer entropy based causality analysis with applications in rooting short-term disturbances for chemical processes.

The transfer entropy (TE) based causality analysis is able to provide a typical solution for fault rooting of industrial processes. However, short-term disturbances that occur during nominal operations of chemical processes are usually neglected because of the fixed time window of TE for global data distributions. Inspired by the selective attention idea, we propose attention transfer entropy (ATE) that helps to locate prominent targets. Concerning temporal features of industrial time series, prior knowledge is employed for constructing an interpretable model. We verify the reliability and effectiveness of the method with coal gasification process data. Additionally, the algorithm is compared to conventional causality analysis methods, proving that ATE enjoys excellent performances in rooting short-term disturbances with lower calculation burden.

Formation and removal of 1,N6-dimethyladenosine in mammalian transfer RNA.

RNA molecules harbor diverse modifications that play important regulatory roles in a variety of biological processes. Over 150 modifications have been identified in RNA molecules. N6-methyladenosine (m6A) and 1-methyladenosine (m1A) are prevalent modifications occurring in various RNA species of mammals. Apart from the single methylation of adenosine (m6A and m1A), dual methylation modification occurring in the nucleobase of adenosine, such as N6,N6-dimethyladenosine (m6,6A), also has been reported to be present in RNA of mammals. Whether there are other forms of dual methylation modification occurring in the nucleobase of adenosine other than m6,6A remains elusive. Here, we reported the existence of a novel adenosine dual methylation modification, i.e. 1,N6-dimethyladenosine (m1,6A), in tRNAs of living organisms. We confirmed that m1,6A is located at position 58 of tRNAs and is prevalent in mammalian cells and tissues. The measured level of m1,6A ranged from 0.0049% to 0.047% in tRNAs. Furthermore, we demonstrated that TRMT6/61A could catalyze the formation of m1,6A in tRNAs and m1,6A could be demethylated by ALKBH3. Collectively, the discovery of m1,6A expands the diversity of RNA modifications and may elicit a new tRNA modification-mediated gene regulation pathway.

Life Table Construction under Different Temperatures and Insecticide Susceptibility Analysis of Uroleucon formosanum (Hemiptera: Aphididae).

Uroleucon formosanum is an important aphid pest of lettuce, but basic information on its biology is scarce. In this study, effects of three constant temperatures (17, 21, and 25 °C, simulating the mean temperature range in greenhouses) on the development and fecundity of U. formosanum were analyzed by constructing a life table. U. formosanum could develop and reproduce under all three temperatures, but the survival rate, development, and fecundity of U. formosanum were affected by temperature. The intrinsic rate of increase was lowest at 17 °C (0.17) and it was significantly less than at 21 °C (0.20) and 25 °C (0.23). Furthermore, U. formosanum had the lowest finite rate of increase (1.19) and the largest mean generation time (20.21) at 17 °C. These results mean that U. formosanum is less adapted to the lower temperatures (17 °C) among these three set temperatures. To screen insecticides for control, susceptibility of U. formosanum to six insecticides including chlorpyrifos, abamectin, beta-cypermethrin, imidacloprid, nitenpyram, and thiamethoxam was evaluated. U. formosanum was relatively sensitive to all six test insecticides. Chlorpyrifos had the highest toxicity to U. formosanum (LC50 = 3.08 mg/L). These data may help to develop integrated management strategies for better population control of U. formosanum.

Broadband efficient anomalous reflection using an aggressively discretized metasurface.

Aggressive discretization in metasurface design-using the least number of unit cells required-can dramatically decrease the phase coverage requirement, thus allowing the use of simple structure and avoiding unit cells with strong resonance, leading to a simple design with broadband performance. An aggressively discretized metasurface with two unit cells per period can realize efficient anomalous reflection. In this work, we investigate the power efficiency and bandwidth of an aggressively discretized metasurface featuring anomalous reflection. Through spectral domain considerations, we find that the theoretical upper limit for the bandwidth of this metasurface reflecting all the incident power into the desired mode is 67%. With aggressive discretization, we design a metasurface with a simple unit cell structure. By tuning the two unit cells, we achieve a metasurface design that reflects more than 80% of the incidence power into the desired anomalous reflection mode over a broad bandwidth of 53.6%. Such bandwidth is unprecedented for an anomalous reflection metasurface. Finally, we fabricate and experimentally demonstrate our anomalous reflection metasurface and obtain bandwidth and efficiency performances which agree well with simulation.

Degradation of Bisphenol A by ozonation in a rotating packed bed: Modeling by response surface methodology and artificial neural network.

The ozonation process of Bisphenol A (BPA) in a rotating packed bed (RPB) was modeled by response surface methodology (RSM) and artificial neural network (ANN). Experiments were performed according to the Box-Behnken design, and the interactive effects of various parameters including ozone concentration, pH, rotation speed of RPB and liquid flow rate on BPA degradation efficiency were investigated. Ozone concentration and pH had the most significant interactive effects on BPA degradation efficiency while rotation speed of RPB had no significant interactive effects with other variables. A second order polynomial equation was obtained to predict BPA degradation efficiency. Also, a multi-layered feed-forward ANN model was constructed based on the data of RSM experiments. Six neurons in hidden layer had the highest correlation coefficient (RANN = 0.99158). A comparison between RSM and ANN models suggested that both can accurately predict BPA degradation efficiency (RRSM = 0.99559). The highest BPA degradation efficiency (99.52 %) was achieved under the conditions of ozone concentration of 20 mg L-1, pH of 11, liquid flow rate of 10 L h-1 and rotation speed of RPB of 800 rpm, which was well predicted by the RSM model (99.54 %) and the ANN model (99.82 %). However, the RSM model was slightly better than the ANN model owing to its higher determination coefficient (R2RSM = 0.9912, R2ANN = 0.9827) and lower mean square error (MSERSM = 0.0001684, MSEANN = 0.0003305).

Analysis of Varieties and Characteristics of Plant Tibetan Medicine in The Stainless Crystal Minrror：A Tibetan Materia Medica / 中国实验方剂学杂志

Tibetan medicine is an important part of traditional Chinese medicine with a long history，complete theories and rich contents. As the core of the sustainable development of Tibetan medicine，the reserve of Tibetan medicine is an important strategic resource of the country. The Stainless Crystal Minrror：A Tibetan Materia Medica is based on the classic book Crystal Pearl Materia Medica. Combined with modern Tibetan medicine，it has collected and introduced more than 1 430 Tibetan medicines and equipped with more than 1 200 colorful pictures of medicinal materials，which has important reference value for the research of Tibetan medicines and the establishment of quality standards. This paper starts from three perspectives of "data analysis-plant distribution-Tibetan medicine classification". Based on The Stainless Crystal Minrror：A Tibetan Materia Medica，the database of plant Tibetan medicine was constructed and analyzed statistically，and the correlation between regional distribution，medicinal parts，altitude and medicinal properties of plant Tibetan medicine was explored. The results showed that The Stainless Crystal Minrror：A Tibetan Materia Medica contained 711 plant medicines，involving 127 families and 368 genera. Angiosperms accounted for 94.1%，among which compositae had 34 genera and 74 species，occupying the first place. There are 10 medicinal parts of plant Tibetan medicine，of which 327 species （44.9%） use whole grass.The average altitude of Tibetan medicine is 3 500 m，among which 81.0% grow above 3 000 m.There is a relationship between altitude and medicinal parts and taste Classification of Tibetan medicines is usually based on the classification of the general drug name，Tibetan name，primitive and family. The analysis of plant medicine resource varieties in The Stainless Crystal Minrror：A Tibetan Materia Medica is beneficial to the effective utilization of Tibetan medicine plant resources.

Transoral robotic surgery for treatment of lingual thyroglossal duct cyst / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the feasibility, safety and efficacy of transoral robotic surgery (TORS) in the treatment of lingual thyroglossal duct cyst (LTGDC). Methods: The clinical data of 10 patients with LTGDC treated with TORS in Tongji Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology from May 2017 to November 2020 were analyzed retrospectively,including 6 males and 4 females, aged 5-44 years. The cysts were fully exposed, and resection usually started from the cephalic side of lesions. The range of resection was 3 to 5 mm away from the lesions, and partial hyoid bone was removed if necessary. Intra-operative robotic set-up time,operation time and estimated blood loss,and post-operative local bleeding, dyspnea and recovery time for oral intake were analyzed. SPSS 12.0 software was used for statistical analysis. Results: The cysts in all 10 patients were successfully resected by TORS with da Vinci Si surgical system. The mean robotic set-up and exposure time, operation time, estimated intraoperative blood loss and recovery time for oral intake were (15.5±7.1) min, (17.6±7.4) min, (8.9±6.4)ml and (2.3±2.2)days, respectively. No patient required tracheostomy intra-or post-operatively, and no symptoms of airway obstruction, postoperative bleeding, pharyngeal fistula, hoarseness and neurological impairment occurred after operation. The patients were followed up for 5 to 47 months, with median follow-up time of 17 months, and no recurrence was observed. Conclusion: TORS is safe and feasible for resection of LTGDC, with rapid recovery and low recurrence rate.

Sensitive and Simultaneous Determination of Uridine Thiolation and Hydroxylation Modifications in Eukaryotic RNA by Derivatization Coupled with Mass Spectrometry Analysis.

The discovery of dynamic and reversible modifications in RNA expands their functional repertoires. Now, RNA modifications have been viewed as new regulators involved in a variety of biological processes. Among these modifications, thiolation is one kind of special modification in RNA. Several thiouridines have been identified to be present in RNA, and they are essential in the natural growth and metabolism of cells. However, detection of these thiouridines generally is challenging, and few studies could offer the quantitative levels of uridine modifications in RNA, which limits the in-depth elucidation of their functions. Herein, we developed a chemical derivatization in combination with mass spectrometry analysis for the sensitive and simultaneous determination of uridine thiolation and hydroxylation modifications in eukaryotic RNA. The chemical derivatization strategy enables the addition of easily ionizable groups to the uridine thiolation and hydroxylation modifications, leading up to a 339-fold increase in detection sensitivities of these modifications by mass spectrometry analysis. The limits of detection of these uridine modifications can be down to 17 amol. With the established method, we discovered and confirmed that a new modification of 5-hydroxyuridine (ho5U) was widely present in small RNAs of mammalian cells, expanding the diversity of RNA modifications. The developed method shows superior capability in determining low-abundance RNA modifications and may promote identifying new modifications in RNA, which should be valuable in uncovering the unknown functions of RNA modifications.

Image-matching assisted dual-frequency phase-sensitive optical time domain reflectometry.

An image-matching assisted dual-frequency phase-sensitive optical time domain reflectometry (Φ-OTDR) is proposed and demonstrated. Compared to the conventional dual-frequency Φ-OTDR, which retrieves data via curve matching, the proposed scheme can effectively improve the temporal resolution and measurement precision while keeping the spatial resolution without additional hardware. In the experiments, with a 10 s temporal window, the proposed scheme realized the same measurement precision as the conventional method that used a 40 s window, suggesting a fourfold improvement of temporal resolution. When both used the 10 s temporal window, the measurement error was suppressed from 21.4% to 1.2% in the sensing for a 2 m hot zone at the end of a 90-m fiber.

Incorporating luminescence-concentrating upconversion nanoparticles and DNA walkers into optical tweezers assisted imaging: a highly stable and ultrasensitive bead supported assay.

We incorporate three conceptual components including luminescence-concentrating upconversion nanoparticles, optical tweezers, and DNA walkers into bead carriers to establish a new imaging analysis.

Spectrally Combined Encoding for Profiling Heterogeneous Circulating Tumor Cells Using a Multifunctional Nanosphere-Mediated Microfluidic Platform.

Comprehensive phenotypic profiling of heterogeneous circulating tumor cells (CTCs) at single-cell resolution has great importance for cancer management. Herein, a novel spectrally combined encoding (SCE) strategy was proposed for multiplex biomarker profiling of single CTCs using a multifunctional nanosphere-mediated microfluidic platform. Different cellular biomarkers uniquely labeled by multifunctional nanosphere barcodes, possessing identical magnetic tags and distinct optical signatures, enabled isolation of heterogeneous CTCs with over 91.6 % efficiency and in situ SCE of phenotypes. By further trapping individual CTCs in ordered microstructures on chip, composite single-cell spectral signatures were conveniently and efficiently obtained, allowing reliable spectral-readout for multiplex biomarker profiling. This SCE strategy exhibited great potential in multiplex profiling of heterogeneous CTC phenotypes, offering new avenues for cancer study and precise medicine.

Improving Flow Bead Assay: Combination of Near-Infrared Optical Tweezers Stabilizing and Upconversion Luminescence Encoding.

To enhance signal acquisition stability and diminish background interference for conventional flow bead-based fluorescence detection methods, we demonstrate here an exceptional microfluidic chip assisted platform by integrating near-infrared optical tweezers with upconversion luminescence encoding. For the former, a single 980 nm laser is employed to perform optical trapping and concurrently excite upconversion luminescence, avoiding the fluctuation of the signals and the complexity of the apparatus. By virtue of the favorable optical properties of upconversion nanoparticles (UCNPs), the latter is carried out by employing two-color UCNPs (Er-UCNPs and Tm-UCNPs) with negligible spectral overlaps. With the assistance of the double key techniques, we fabricated complex microbeads referred to a UCNPs-miRNAs-microbead sandwich construct by a one-step nucleic acid hybridization process and then obtained uniform terrace peaks for the automatic and simultaneous quantitative determination of miRNA-205 and miRNA-21 sequences with a detection limit of pM level on the basis of a special home-built flow bead platform. Furthermore, the technique was successfully applied for analyzing complex biological samples such as cell lysates and human tissue lysates, holding certain potential for disease diagnosis. In addition, it is expected that the flow platform can be utilized to investigate many other biomolecules of single cells and to allow analysis of particle heterogeneity in biological fluid by means of optical tweezers.

Development of C60-based labeling reagents for the determination of low-molecular-weight compounds by matrix assisted laser desorption ionization mass spectrometry (II): Determination of thiols in human serum.

Perturbation of thiol homeostasis in biological fluids are thought to be associated with several diseases, and reliable analytical methods for the determination of low molecular weight (LMW) thiols in human plasma or serum are thus required. In this study, a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) method is described for high throughput determination of four LMW thiols (glutathione, cysteine, homocysteine and cysteinylglycine) in human serum. It is based on the use of a bromoacetyl functionalized C60 (Br-C60) as a derivatization reagent to label thiols. The Br-C60 labeling can add an 832-Da tag to thiols, which moves thiol signals to high mass region and effectively avoids the signal interference generated by the traditional MALDI matrix below 800 Da. The labeling can be completed within 5 min under microwave-assisted condition. Thereby, the Br-C60 labeling based MALDI-TOF MS analytical method can achieve high throughput analysis of LMW thiols in serum. Good linearities of the method for the thiols in human serum were obtained in the range of 0.5-500.0 µM with correlation coefficient (R) greater than 0.9960. The limit of detection is in the range of 0.07-0.18 µM for the investigated thiols in human serum with relative standard deviations of lower than 13.5% and recoveries ranging from 81.9 to 117.1%. Using the method, four thiols in microliter serum samples of breast cancer (BC) patients were determined. The result showed that the contents of the four thiols in BC serum samples significantly changed compared to the healthy control (HC).

Chemical labeling - Assisted mass spectrometry analysis for sensitive detection of cytidine dual modifications in RNA of mammals.

RNA molecules carry diverse modifications that exert important influences in many cellular processes. In addition to the single modification occurring in either nucleobase or 2' hydroxyl of ribose in RNA, some dual modifications occur in both the nucleobase and 2' hydroxyl of ribose in RNA. 2'-O-methyl-5-methylcytidine (m5Cm), the dual modifications of cytidine, was first discovered from the tRNA of archaea. Recent studies identified that 2'-O-methyl-5-hydroxymethylcytidine (hm5Cm) and 2'-O-methyl-5-formylcytidine (f5Cm) were present in the anticodon of cytoplasmic tRNA of mammals. Similar to the series of single modification of cytidines of 5-methylcytosine (m5C), 5-hydroxymethylcytidine (hm5C), 5-formylcytidine (f5C), and 5-carboxylcytidine (ca5C) in nucleic acids, the dual modifications of m5Cm, hm5Cm, f5Cm and 2'-O-methyl-5-carboxylcytidine (ca5Cm) may also constitute the series of cytidine modifications in mammals. However, it is normally challenging to detect these modifications because of their low endogenous levels. Here, we established a method by chemical labeling-assisted liquid chromatography - electrospray ionization - tandem mass spectrometry (LC-ESI-MS/MS) analysis for the sensitive and simultaneous determination of all these four cytidine dual modifications, i.e., m5Cm, hm5Cm, f5Cm and ca5Cm. Three different labeling reagents (2-bromo-1-(3,4-dimeth oxyphenyl)-ethanone, BDMOPE; 2-bromo-1-(4-methoxyphenyl)-ethanone, BMOPE; 2-bromo-1-(4-diethylaminophenyl)-ethanone, BDEPE) were used for the chemical labeling. The results showed that the detection sensitivities of m5Cm, hm5Cm, f5Cm and ca5Cm increased up to 462 folds after chemical labeling. With the developed method, we achieved the simultaneous detection of m5Cm, hm5Cm and f5Cm in RNA of mammals. In addition, we found these cytidine dual modifications mainly exist in small RNA (<200 nt) and barely detected in other types of RNA. Moreover, we found that the levels of m5Cm in RNA of human lung carcinoma tissues significantly increased, while hm5Cm and f5Cm significantly decreased compared to tumor adjacent normal tissues. The significant changes of m5Cm, hm5Cm and f5Cm levels may serve as indicator for the detection and prognosis of lung cancer.

Self-assembly of MoO3-decorated carbon nanofiber interlayers for high-performance lithium-sulfur batteries.

Lithium-sulfur (Li-S) batteries are attractive for next-generation energy storage systems due to their high theoretical capacity and energy density. However, the undesired shuttling of soluble lithium polysulfides (Li2Sn, 2 < n≤ 8) and the uncontrolled growth of lithium dendrites have hindered their practical applications. Herein, a self-assembled freestanding MoO3/carbon nanofiber (MoO3/CNF) composite membrane is effectively integrated into Li-S batteries as a functional interlayer. Improved cell performance is achieved due to the strong interfacial chemical and physical interactions between the interlayers with Li2Sn. The Li-S batteries exhibit a decent cyclic stability with a fading rate of 0.12% per cycle for 500 cycles at 1675 mA g-1, a high rate performance and a low self-discharge rate. In this rational design, the CNF network provides abundant electron pathways and physically prevents polysulfide diffusion. The polar MoO3 nanorods act as effective anchoring sites by the chemical interactions with Li2Sn. Meanwhile, the suppressed Li-dendrite growth on the Li-anode surface results in a stable Li stripping/plating.

Diazo Reagent Labeling with Mass Spectrometry Analysis for Sensitive Determination of Ribonucleotides in Living Organisms.

Ribonucleotide analogues and their related phosphorylated metabolites play critical roles in tumor metabolism. However, determination of the endogenous ribonucleotides from the complex biological matrix is still a challenge due to their high structural similarity and high polarity that will lead to the low retention and low detection sensitivities by liquid chromatogram mass spectrometry analysis. In this study, we developed the diazo reagent labeling strategy with mass spectrometry analysis for sensitive determination of ribonucleotides in the living organism. A pair of light and heavy stable isotope labeling reagents, 2-(diazomethyl)-N-methyl-N-phenyl-benzamide (2-DMBA) and d5-2-(diazomethyl)-N-methyl-N-phenyl-benzamide (d5-2-DMBA), were synthesized to label ribonucleotides. 2-DMBA showed high specificity and high efficiency for the labeling of ribonucleotides. Our results demonstrated that the detection sensitivities of 12 ribonucleotides increased by 17-174-fold upon 2-DMBA labeling. The obtained limits of detection (LODs) of ribonucleotides ranged from 0.07 fmol to 0.41 fmol. Using this method, we achieved the sensitive and accurate detection of ribonucleotides from only a few cells (8 cells). To the best of our knowledge, this is the highest detection sensitivity for ribonucleotides ever reported. In addition, we found that the contents of almost all of these ribonucleotides were significantly increased in human breast carcinoma tissues compared to tumor-adjacent normal tissues, suggesting that endogenous ribonucleotides may play certain functional roles in the regulation of cancer development and formation. This method also can be potentially applied in the analysis of phosphorylated compounds.