Analysis on acupoint selection rules in the treatment of metabolism-associated fatty liver disease based on data mining. / åŸºäºŽæ•°æ®æŒ–æŽ˜æŠ€æœ¯åˆ†æžé’ˆç¸æ²»ç–—ä»£è°¢ç›¸å ³è„‚è‚ªæ€§è‚ç— çš„é€‰ç©´è§„å¾‹.

OBJECTIVES: To explore the rules of acupoint selection in the treatment of metabolic-associated fatty liver disease (MAFLD) with acupuncture and moxibustion by using data mining technology. METHODS: The clinical research literature on acupuncture treatment of MAFLD was collected from PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure, Wanfang Database, VIP Database and China Biology Medicine from their inception to November 20, 2022. According to our inclusion and exclusion criteria, the literature was independently screened and re-screened by two research members, and the screened results were checked, followed by establishing an acupoint prescription database using Excel 2019. Descriptive statistics of acupoints applied frequency, involved meridians, locations and specific acupoints were perpormed. Then, SPSS Modeler18.0 software was used to conduct analysis about association rules, and the SPSS Statistics 26.0 software was used to perform cluster analysis on high-frequency acupoints, exploring the characteristics and rules of acupoint selection and combination in the treatment of MAFLD. RESULTS: Totally, 178 papers were collected, containing 130 acupoints, with a total application frequency of 1 305. The top five acupoints are Zusanli (ST36), Fenglong (ST40), Ganshu (BL18), Taichong (LR3) and Sanyinjiao (SP6). The commonly involved meridians are the Stomach Meridian of Foot Yangming, Bladder Meridian of Foot Taiyang, and Spleen Meridian of Foot Taiyin. The employed acupoints are mostly located in the lower limbs and abdomen, and the five Shu acupoints and crossing acupoints are in the majority. The association rule analysis of high frequency acupoints indicated that of the 16 qualified acupoint groups, the top 5 with close correlation degrees are ST36 and ST40, ST36 and LR3, ST36 and SP6, ST40 and LR3 and ST36, ST36 and SP6 and ST40. Further, 3 effective clusters were obtained by cluster analysis. CONCLUSIONS: Acupuncture and moxibustion treatment of MAFLD follows the therapeutic principles of soothing the liver, invigorating the spleen, tonifying the kidney, and resolving phlegm and removing dampness. The core acupoint group is ST36, ST40 and LR3, and the combination of acupoints is based on syndrome differentiation. These results may provide a useful reference for clinical practice.

A Data Mining Study for Analysis of Acupoint Selection and Combinations in Acupuncture Treatment of Carpal Tunnel Syndrome.

Background: Carpal tunnel syndrome (CTS) is the most prevalent upper limb compressive neuropathy. A considerable number of clinical trials and meta-analyses have provided evidence supporting the effectiveness of acupuncture in treating CTS. Nevertheless, the ideal choice of acupoints remains ambiguous. Objective: A data mining analysis was conducted with the objective of determining the most effective acupoint combinations and selection for CTS. Methods: A search was conducted across seven Chinese and English electronic bibliographic databases spanning from their inception to March 2023. Selected were clinical trials that evaluated the efficacy of acupuncture therapy for CTS, with or without randomised controlled methods. Data extraction mainly included acupoint prescriptions. Information such as first author, study design and study setting were also extracted. The principal outcomes comprised the clinical manifestations linked to CTS. Statistical descriptions were generated using Excel 2019. The analysis of association rules was conducted using SPSS Modeler 18.0. Using SPSS Statistics 26.0, exploratory factor analysis and cluster analysis were conducted. Results: 142 trials (including 86 RCTs and 56 non RCTs) were identified, and 193 groups of effective prescriptions involving 68 acupoints were extracted. The most frequently used acupoints were Da-ling (PC7), Nei-guan (PC6), He-gu (LI4), Wai-guan (TE5), and Yang-xi (LI5). The most frequently used meridians were the pericardial meridian and the large intestine meridian. The majority of special acupoints used were Five-shu points and Yuan-source points, with acupoints on the upper limbs being the most frequently used. The core acupoint groups were analyzed and 11 groups of association rules, 8 factors, and 5 effective cluster groups were obtained. Conclusion: The evidence-based acupoint selection and combinations of acupuncture therapy for carpal tunnel syndrome were provided by the findings of this study.

Acupoint selection rules of acupuncture and moxibustion in treating neurogenic bladder based on data mining. / 基于数据挖掘技术分析针灸治疗神经源性膀胱的选穴规律.

OBJECTIVES: To explore the rules of acupoint selection in the treatment of neurogenic bladder (NB) with acupuncture and moxibustion by using data mining. METHODS: The clinical research literatures on acupuncture treatment of NB were collected from PubMed, Embase, Cochrane Library, CNKI, Wanfang Database, VIP Database and China Biology Medicine from retrieved to January 1, 2023. The acupoint prescription database was established using Excel 2019. SPSS Modeler 18.0 and SPSS Statistics 26.0 softwares were used to conduct the frequency, meri-dians, locations, specific acupoints analysis and association rules analysis, factor analysis, cluster analysis, etc., to explore the characteristics and rules of acupoint selection in acupuncture and moxibustion treatment of NB. RESULTS: Totally 313 papers were included, including 110 acupoints with a total frequency of 1 995. The high-frequency acupoints are Zhongji (CV3), Guanyuan (CV4), Sanyinjiao (SP6), etc. The commonly used meridians are the Bladder Meridian of Foot Taiyang and Conception Vessel. The involved acupoints are mostly located in the lumbosacral region and abdomen, and intersection acupoints, mu-front acupoints and back-shu acupoints are the majority in the specific acupoints. The core acupoints group was analyzed, and 17 groups of association rules, 7 factors and 6 effective cluster groups were obtained. CONCLUSIONS: Acupuncture and moxibustion treatment of NB follows the therapeutic principles of toni-fying the kidney, invigorating the spleen, and soothing the liver. The core acupoints group is CV3-CV4-SP6.

By Carrot or by Stick: The Influence of Encouraging and Discouraging Facial Feedback on Implicit Rule Learning.

Implicit learning refers to the process of unconsciously learning complex knowledge through feedback. Previous studies investigated the influences of different types of feedback (e.g., social and non-social feedback) on implicit learning. This study focused on the social information presented in the learning situation and tried to explore the effects of different social feedback on implicit rule learning. We assigned participants randomly into an encouraging facial feedback group (happy expression for correct answer, neutral but not negative expression for incorrect answer) and a discouraging facial feedback group (neutral but not happy expression for correct answer, negative expression for incorrect answer). The implicit learning task included four difficulty levels, and social feedback was presented in the learning phase but not the testing phase in two experiments. The only difference between the two experiments was that the sad face used as negative feedback in Experiment 1 was replaced with an angry face in Experiment 2 to enhance the ecological validity of the discouraging facial feedback group. These two experiments yielded consistent results: the performances in the encouraging facial feedback group were more accurate in both the learning and the testing phases at all difficulty levels. These findings indicated that the influence of encouraging social feedback for a better implicit learning achievement was stable and established a new groundwork for future research on incentive-based education, making it critical to investigate the impact of various forms of encouraging-based education on learning.

Research Trends of Acupuncture Therapy for Chronic Pain-Related Depression or Anxiety from 2003 to 2023: A Bibliometric Analysis.

Background: Chronic pain (CP) as a long-lasting stressor can often cause mood disorders, such as depression or anxiety. The comorbidity of CP and mood disorders poses challenges for treatment and increases healthcare costs. Acupuncture has emerged as a widely utilized approach to alleviate both CP and mood disorders. However, there is a lack of well-established bibliometric analyses in this area of research. Therefore, this study aimed to identify the current hotspots and research trends regarding the use of acupuncture for treating CP-related depression or anxiety. Methods: We searched the Web of Science Core Collection spanning from 2003 to 2023 to identify relevant literature about the use of acupuncture for treating CP-related depression or anxiety. Bibliometric and visualization analyses were performed using CiteSpace 5.7.R5 and Vosviewer 1.6.19 software. Results: A total of 254 articles published between 2003 and 2023 were included, revealing an upward trajectory with some fluctuations in publication numbers over the past two decades. China and the Beijing University of Chinese Medicine were the most productive country and institution in this field. Fang JQ and Vickers AJ ascended as the most prolific and influential authors, respectively. Trials was the journal with the highest number of publications, while Pain and BMJ-British Medical Journal exhibited the highest citation and centrality, respectively. "Acupuncture", "depression", and "chronic pain" were the top three keywords. The hotspots in this domain encompass types of chronic pain that predispose to negative mood, including fibromyalgia, irritable bowel syndrome (IBS), and neuropathic pain. Randomized controlled trials (RCTs), mechanisms, and evidence-based evaluations are the main research directions. Conclusion: This study uses bibliometric techniques to analyze the research hotspots and forefronts of acupuncture as a therapeutic approach for CP-related depression or anxiety. Our objective is to provide researchers with valuable references and identify research focal points for future investigations.

A bibliometric and visualization analysis of global research on postherpetic neuralgia from 2000 to 2022: A review.

Postherpetic neuralgia (PHN) represents a notable clinical challenge as it is the most prevalent and severe complication of herpes zoster (HZ). The primary objective was to investigate the current research status and hotspots of PHN research during the period from 2000 to 2022. The literature pertaining to PHN was gathered through the utilization of the Web of Science Core Collection, spanning from January 2000 to December 2022. The software, CiteSpace version 6.2.R2, was employed to produce visual depictions of publications related to PHN across various dimensions such as year, country/region, institution, journal, author, keyword, and reference. This study involved a total of 3505 papers. The USA held a dominant position in the production of scholarly articles. Argentina exhibited the highest frequency of participation in international collaboration. Out of all the institutions, Pfizer exhibited the highest degree of productivity. Harvard University exhibited the highest frequency of participation in international collaboration. The Pain exhibited the most noteworthy productivity rate and citation count among all other journals. Ralf Baron was identified as the most productive author, whereas DWORKIN RH attained the highest citation count. Contemporary scholarly investigations are predominantly centered on identifying risk factors, devising preventative measures, and exploring novel and secure methods of pain management. The current investigation has revealed the focal areas and patterns of studies pertaining to PHN. Presently, the research in this field is focused on identifying the risk factors and preventive measures for PHN, alongside exploring novel and secure pain management strategies.

Electroacupuncture promotes synaptic plasticity in rats with chronic inflammatory pain-related depression by upregulating BDNF/TrkB/CREB signaling pathway.

BACKGROUND: Chronic inflammatory pain (CIP) frequently coincides with depression among patients. The onset and development of pain and depression are associated with altered neural synaptic plasticity. Electroacupuncture (EA) can effectively relieve CIP and depression. However, the underlying mechanisms have not been fully illustrated. OBJECTIVE: To explore whether EA can relieve CIP and depression by regulating hippocampal synaptic plasticity, and the present study offers foundational evidence for the efficacy of EA in treating CIP-related depression (CIPD). METHODS: Rats were divided into four groups: 0.9% normal saline group, complete Freund's adjuvant (CFA) group, CFA + duloxetine group, and CFA + EA group. Pain hypersensitivity was detected by mechanical withdrawal threshold and thermal paw withdrawal latency, and the depression level was gauged using the open field test, the sucrose preference test, and the forced swimming test. The morphology of the hippocampal neurons was observed using Nissl staining. The protein expression levels of synuclein (Syn), postsynaptic density protein-95 (PSD-95), brain-derived neurotrophic factors (BDNFs), tyrosine-protein kinase B (TrKB), p-TrkB, cAMP response element binding protein (CREB), and p-CREB were measured by western blotting and immunofluorescence staining. BDNF and TrkB mRNA expression were detected using quantitative real-time polymerase chain reaction (PCR) (qRT-PCR). The content of 5-hydroxytryptamine (5-HT) and γ-aminobutyric acid (GABA) was detected using enzyme-linked immunosorbent assay, and the glutamic acid (Glu) content was determined using the ultraviolet colorimetry method. The hippocampal neuron ultrastructure was observed using transmission electron microscopy. RESULTS: EA could alleviate CIP and related depressive behaviors as well as protect the hippocampal neuronal structure from damage and regulate 5-HT/GABA/Glu levels in the hippocampus. Additionally, EA could significantly increase the expression of synapse-associated proteins such as PSD-95 and Syn by activating the BDNF/TrKB/CREB signaling pathway. CONCLUSION: EA improves pain and depressive behaviors in CIPD rats, and the mechanism may be related to synaptic plasticity mediated by the BDNF/TrKB/CREB signaling pathway.

Analysis of Acupoint Selection and Combinations in Acupuncture Treatment of Piriformis Syndrome: A Protocol for Data Mining.

Background: Piriformis syndrome (PS) is a neuromuscular condition characterized by discomfort in the gluteal region. The efficacy of acupuncture as a treatment modality for PS has been substantiated through a multitude of clinical trials. However, certain queries persist, such as the optimal approach for identifying the most efficacious acupoints. The objective of this study is to perform an initial data mining analysis aimed at identifying the optimal acupoint selection and combinations for the treatment of PS. Methods: We will search 7 electronic bibliographic databases (PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure, Wanfang Database, Chinese Biomedical Literature Database and Chongqing VIP Database) from inception to June 2023. We will select clinical trials that evaluate the efficacy of acupuncture therapy in the management of PS. Exclusions will be made for reviews, protocols, animal trials, case reports, systematic reviews, and meta-analyses. The primary outcome measure will be clinical outcomes associated with PS. Descriptive statistics will be performed in Excel 2019. Association rule analysis will be performed in SPSS Modeler 18.0. Exploratory factor analysis and cluster analysis will be performed in SPSS Statistics 26.0. Results: This study will investigate the most effective acupoint selection and combinations for patients with PS. Conclusion: Our findings will provide evidence for the effectiveness and potential treatment prescriptions of acupoint application for patients with PS, helping clinicians and patients make a more informed decision together.

Acupuncture Methods for Piriformis Syndrome: A Protocol for Systematic Review and Network Meta-Analysis.

Background: Piriformis syndrome (PS) is a common disease that is difficult to clinically treat, which seriously affects people's health and living quality. Multiple clinical studies have demonstrated that acupuncture therapy is an effective treatment for PS, but the optimal method of various acupuncture methods has remained unknown. The aim of this study protocol is to compare the efficacy and the safety of different acupuncture methods for PS. Methods: We will search seven electronic bibliographic databases (PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure, Wanfang Database, Chinese Biomedical Literature Database, and Chongqing VIP Database) from inception to June 2023. Randomized controlled trials (RCTs) will be selected for assessing the effectiveness of acupuncture therapy on PS. Primary outcome measure for this study is pain intensity, and the secondary outcomes are clinical symptoms and signs, physical activity, effective rate, and adverse events. Study selection, data extraction, and risk of bias assessment will be independently completed by at least two reviewers. Then, standard pairwise meta-analysis and network meta-analysis will be conducted using Revman 5.3, Stata V.15.1 and OpenBUGS V.3.2.3. The heterogeneity will be explored by subgroup, meta-regression, and sensitivity analysis, if applicable. Finally, Grading of Recommendations, Assessment, Development and Evaluation (GRADE) pro system will be used to evaluate the quality of evidence. Results: Our systematic review and network meta-analysis will identify the best acupuncture treatment method for PS patients. Conclusion: The results of our review will help decision-makers make educated choices regarding evidence-based acupuncture management options for PS. Prospero Registration Number: No. CRD42022335028.

Electroacupuncture attenuates chronic inflammatory pain and depression comorbidity by inhibiting hippocampal neuronal apoptosis via the PI3K/Akt signaling pathway.

In chronic inflammatory pain (CIP) and depression, neuroapoptosis has been identified as a contributing factor. Electroacupuncture (EA) shows promise as an alternative therapy for this comorbidity. However, the underlying mechanism remains unclear. This study aimed to investigate the effects of EA on hippocampal neuronal apoptosis in rats with CIP and depression. Rats received plantar injections of complete Freund's adjuvant (CFA) on days 0 and 14. They were then divided into groups: sham operation, model, EA, and duloxetine. EA was administered at Hegu (LI4) and Taichong (LR3) from days 15 to 28, while the duloxetine group received duloxetine and distilled water daily (0.1 mg/ml). Pain behavior was assessed using the mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) tests. Depression-like behavior was evaluated through the sucrose preference test (SPT), open-field test (OFT), and forced swim test (FST). Hematoxylin and eosin (HE) staining was employed to assess pathological changes in the hippocampus. Nerve cell apoptosis was determined using TUNEL fluorescence staining. Western blot analysis was conducted to measure the protein expression of Bcl-2, Bax, p-PI3K/PI3K, and p-Akt/Akt. EA demonstrated significant pain intensity reduction and alleviation of pain-related depressive symptoms. Our findings from the HE staining confirmed that CIP induced by CFA led to morphological changes in the hippocampus, while EA effectively reversed these pathological alterations. Moreover, EA intervention remarkably reduced neuronal apoptosis and exhibited an upregulation of Bcl-2 protein expression accompanied by a decrease in Bax expression. Additionally, EA activated the PI3K/Akt signaling pathway. Overall, our study suggests that EA holds the potential to improve pain and depressive behaviors in rats with CIP and depression comorbidity, potentially mediated through the activation of the PI3K/Akt pathway, leading to a reduction in hippocampal neuronal apoptosis.

Analysis of Acupoint Selection and Combinations in Acupuncture Treatment of Carpal Tunnel Syndrome: A Protocol for Data Mining.

Background: Carpal tunnel syndrome (CTS), as the most common compression neuropathy in the upper limb, can lead to upper limb dysfunction in patients. The effectiveness of acupuncture in treating CTS has been validated based on numerous clinical trials and meta-analyses, but questions remain, such as how to select the best acupoints. Our purpose is to conduct the first data mining analysis to identify the most effective acupoint selection and combinations for treating CTS. Methods: We will search 7 electronic bibliographic databases (PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure, Wanfang Database, Chinese Biomedical Literature Database and Chongqing VIP Database) from inception to March 2023. Clinical trials assessing the effectiveness of acupuncture therapy on the management of CTS will be selected. Reviews, protocols, animal trials, case reports, systematic reviews, and meta-analyses will be excluded. The primary outcome measure will be clinical result associated with CTS. Descriptive statistics will be performed in Excel 2019. Association rule analysis will be performed in SPSS Modeler 18.0. Exploratory factor analysis and cluster analysis will be performed in SPSS Statistics 26.0. Results: This study will investigate the most effective acupoint selection and combinations for patients with CTS. Conclusion: Our findings will provide evidence for the effectiveness and potential treatment prescriptions of acupoint application for patients with CTS, helping clinicians and patients make a more informed decision together.

Circular RNA hsa_circ_0103552 Promotes Proliferation, Migration, and Invasion of Breast Cancer Cells through Upregulating Cysteine-Rich Angiogenic Inducer 61 (CYR61) Expression via Sponging MicroRNA-515-5p.

Circular RNAs (circRNAs) exert a significant regulatory function on tumor progression. This work intends to probe into the biological function and regulatory mechanism of circRNA_0103552 (circ_0103552) in breast cancer carcinogenesis. In this study, circ_0103552, microRNA-515-5p (miR-515-5p), and cysteine-rich angiogenic inducer 61 (CYR61) mRNA expressions in breast cancer cells and tissues were determined by quantitative real-time polymerase chain reaction, followed by cell counting kit 8 and Transwell experiments to examine the multiplication, migration, and invasion of breast cancer cells. Circular RNA Interactome database and StarBase database were searched, and dual-luciferase reporter gene experiments were applied to verify the targeting relationship between circ_0103552 and miR-515-5p, and between miR-515-5p and CYR61, and Western blot was adopted to the regulatory function of circ_0103552 and miR-515-5p on CYR61 protein expression. Circ_0103552 expression was found to be remarkably up-modulated in breast cancer tissues and cells, and circ_0103552 overexpression facilitated the multiplication, migration, and invasion of breast cancer cells, while knocking down circ_0103552 induced the opposite effects. Mechanistically, circ_0103552 could sponge miR-515-5p and restrained its expression in breast cancer cells. MiR-515-5p could counteract the functions of circ_0103552 in breast cancer cells. Additionally, CYR61 was revealed to be a downstream target of miR-515-5p in breast cancer cells. In summary, this study shows that circ_0103552 up-modulates CYR61 expression by targeting miR-515-5p and thus facilitates the multiplication, migration, and invasion of breast cancer cells.

Determining Essential Requirements for Fluorophore Selection in Various Fluorescence Applications Taking Advantage of Diverse Structure-Fluorescence Information of Chromone Derivatives.

Herein, we report our work exploring the essential requirements for fluorophore selection during the development of various fluorescence applications. We assembled a library of chromone-derived fluorophores with diverse structure-fluorescence properties, which allowed us to choose the fluorophore pairs with similar structures but differing fluorescence properties and compared the performance of the selected fluorophore pairs in three types of commonly used fluorescence applications. We found that the selection standard of a suitable fluorophore is variable depending on the application. (1) In fluorescence imaging, fluorophores with strong and constant fluorescence under various conditions, such as a large pH range, are preferred. Notably, (2) in the detection of bioactive species, fluorophores with relatively lower fluorescence quantum yield favor the detection sensitivity. Furthermore, (3) in enzymatic assays employing fluorescence, the key parameter is the binding affinity between the fluorophore and the enzyme.

Analysis of Circulating Tumor DNA to Predict Neoadjuvant Therapy Effectiveness and Breast Cancer Recurrence.

PURPOSE: Real-time detection and intervention can be used as potential measures to markedly decrease breast cancer mortality. Assessment of circulating tumor DNA (ctDNA) may offer great benefits for the management of breast cancer over time. However, the use of ctDNA to predict the effectiveness of neoadjuvant treatment and recurrence of breast cancer has rarely been studied. METHODS: We prospectively recruited 31 breast cancer patients with 4 subtypes. Three time points were set in this study, including before any therapy (C1), during surgery (T), and six months after surgery (C2). We collected peripheral blood samples from all 31 patients at C1, tumor tissue from all 31 patients at T, and peripheral blood samples from 25 patients at C2. Targeted 727-gene panel sequencing was performed on ctDNA from all blood samples and tissue DNA from all tissue samples. Somatic mutations were detected and analyzed using a reference standard pipeline. Statistical analysis was performed to identify possible associations between ctDNA profiles and clinical outcomes. RESULTS: In total, we detected 159, 271, and 70 somatic mutations in 30 C1 samples, 31 T samples, and 12 C2 samples, respectively. We identified specific genes, such as PIK3CA, TP53, and KMT2C, which were highly mutated in the tissue samples. Furthermore, mutated KMT2C observed in ctDNA of the C2 samples may be an indicator of breast cancer recurrence. CONCLUSION: Our study highlights the potential of ctDNA analysis at different timepoints for assessing tumor progression and treatment effectiveness, as well as prediction of breast cancer recurrence.

Surgery on primary melanoma of the breast.

Malignant melanoma of the breast (MMB) is a rare disease that accounts for 3-5% of all melanomas. It can be expressed as a metastatic mammary gland or primary mammary melanin of the mammary gland. Primary MMB (PMMB) can be divided into two types: parenchymal melanoma without skin involvement and melanoma involving skin on the breast. The diagnosis of PMMB relies mainly on histopathological diagnosis. In this diagnostic approach, S100 and HMB45 have a higher specificity in the diagnosis of melanoma immunohistochemistry. At the same time, mutations in the BRAF (V600E) gene have further aided the diagnosis of PMMB. After the diagnosis, the main treatment is mainly based on surgery. The main surgical methods include breast-conserving surgery and mastectomy. With advances in technology and surgical skills, the combination of patient aesthetic satisfaction and tumor safety is the goal of modern breast surgeons. This article reviews the current status of PMMB surgical treatment.

Quantitative Evaluation of in Vivo Target Efficacy of Anti-tumor Agents via an Immunofluorescence and EdU Labeling Strategy.

Current methods used to evaluate in vivo target efficacy of selected compound include western blot to semi-quantitatively analyze protein expression. However, problems arise as it is difficult to compare in vivo target efficacy of anti-tumor agents with the same mode of action. It is therefore desirable to develop a protocol that can quantitatively display in vivo target efficacy while also providing other useful information. In this study EdU labeling was used to mark out the proliferating area. The tumor tissue was accordingly divided into proliferating and non-proliferating areas. Fifteen tumor related proteins were stained by immunofluorescence and were found to express in either the proliferating or non-proliferating areas. This allows the quantitative analysis of protein expressions within the precise area. With simple image analysis, our method gave precise percent changes of protein expression and cell proliferation between the drugs treated group and the control group. Additional information, such as, the status of protein expression can also be obtained. This method exhibits high sensitivity, and provides a quantitative approach for in vivo evaluation of target efficacy.

Exploration of the Fluorescent Properties and the Modulated Activities against Sirtuin Fluorogenic Assays of Chromenone-Derived Natural Products.

Chromenone-derived natural products include chromones (flavone, isoflavone) and coumarins. Chromenone compounds not only exhibit impressive biological activities, but also are an important resource of experimentally used fluorophores, such as, 7-amino-4-methylcoumarin (AMC). Various chromenone compounds have reported to have weak fluorescence, and this has the potential to interfere with the measurements during AMC fluorogenic assays and result in non-robust assay readouts. Several flavones and isoflavones were found as SIRT1 activators, while fluorogenic sirtuin assays utilized AMC labelled peptides as the substrates. In this study we investigated whether the fluorescent properties of chromenone-derived natural products interrupt the measurement of SIRT1/2 modulated activities. We found that the reported SIRT1 activators: flavones were detected with the SIRT1 activation activity, but isoflavones were not detected with SIRT1 activation activity, and instead that they were found to be fluorogenic compounds. Another chromenone compound, osthole, exhibited a moderate SIRT2 inhibitory activity with an IC50 of 10 µM. In conclusion, the fluorescent properties of these chromenone compounds do affect the measurement of the sirtuin activities of both inhibitors and activators. However, if the possible fluorescence properties are mitigated in the assay readout, these fluorogenic assays enable the screening of activity modulators.

A Suzuki-Miyaura method for labelling proliferating cells containing incorporated BrdU.

The 5-bromo-2'-deoxyuridine (BrdU) incorporation cell proliferation assay is the most commonly used method for assessing DNA replication. The current detection of BrdU in cells relies on antibody immunostaining, but has various limitations including low antibody specificity and poor tissue penetration. In this study, we utilised a Suzuki-Miyaura reaction to develop a chemical method to label cellular BrdU with fluorescent boronic acid probes. The coupling conditions were optimised for complex cellular environments, and the key observation was the need to use oxygen scavengers and zerovalent palladium to prevent side reactions and increase the rate of coupling. The reliability and specificity of the BrdU Suzuki-Miyaura labelling method were verified under various biological conditions. The applicability of the BrdU Suzuki-Miyaura labelling methodology was also investigated, and we show that labelling cellular BrdU is highly sensitive and reliable, which is comparable to the ideal performance of BrdU immunostaining. Moreover, the Suzuki-Miyaura reaction protocol provides high BrdU recognition specificity. Taken together, the BrdU Suzuki-Miyaura labelling protocol provides an attractive alternative to the more traditional cell proliferation assay.

Expression, purification, and characterization of a novel acidic Lipoxygenase from Myxococcus xanthus.

The gene encoding a novel acidic lipoxygenase from Myxococcus xanthus DK1622 (accession: WP_011551853.1) was cloned into vector pET-28a and expressed in Escherichia coli BL21(DE3). The recombinant enzyme (rMxLOX), with a molecular weight of approximately 80 kDa, was purified to homogeneity using one-step nickel-affinity chromatography and showed an activity of 5.6 × 104 U/mg. The optimum pH and temperature for rMxLOX activity were found to be 3.0 and 30 °C, respectively. Purified rMxLOX exhibited activity towards linoleic acid and arachidonic acid as substrates, with linoleic acid being the better substrate (Km and kcat values of 0.048 mM and 13.3/s, respectively). The synthetic dye aniline blue was decolorized 69.7 ± 3.5%, following incubation with rMxLOX for 35 min. These results reveal the potential for the use of rMxLOX in the pulp, textile, and wastewater treatment industries.