DNAzyme-RCA-based colorimetric and lateral flow dipstick assays for the point-of-care testing of exosomal m5C-miRNA-21.

Methylation of microRNAs (miRNAs) is a post-transcriptional modification that affects miRNA activity by altering the specificity of miRNAs to target mRNAs. Abnormal methylation of miRNAs in cancer suggests their potential as a tumor marker. However, the traditional methylated miRNA detection mainly includes mass spectrometry, sequencing and others; complex procedures and reliance on large instruments greatly limit their application in point-of-care testing (POCT). Based on this, we developed DNAzyme-RCA-based gold nanoparticle (AuNP) colorimetric and lateral flow dipstick (LFD) assays to achieve convenient detection of exosomal 5-methylcytosine miRNA-21 (m5C-miRNA-21) for the first time. The two assays achieved specific recognition and linear amplification of m5C-miRNA-21 through the DNAzyme triggered RCA reaction and color output with low background interference through AuNP aggregation induced by base complementary pairing. The lowest concentration of m5C-miRNA-21 visible to the naked eye of the two assays can reach 1 pM and 0.1 pM, respectively. Detection of exosomal m5C-miRNA-21 in clinical blood samples showed that the expression level of m5C-miRNA-21 in colorectal cancer patients was significantly higher than that in healthy individuals. This approach not only demonstrates a new strategy for the detection of colorectal cancer but also provides a reference for the development of novel diagnostic tools for other miRNA methylation-related diseases.

Effects of Dl-3-n-butylphthalide on neurological function, hemodynamics and Hcy concentration in cerebral hemorrhage: a systematic review and meta-analysis.

Background: Dl-3-n-Butylphthalide (NBP) has emerged as a potential therapeutic agent for cerebral hemorrhage, despite not being included in current guideline recommendations. Investigating the underlying physiological and pathological mechanisms of Dl-3-n-Butylphthalide in cerebral hemorrhage treatment remains a critical area of research. Objective: This review aims to evaluate the efficacy of Dl-3-n-Butylphthalide in cerebral hemorrhage treatment and elucidate its potential biological mechanisms, thereby providing evidence to support treatment optimization. Methods: A comprehensive search of seven electronic databases (PubMed, Web of Science, Embase, Cochrane Library, China National Knowledge Infrastructure, VIP, and Wanfang Database) was conducted for studies published up to September 2023. Screening and data extraction were performed by a team of researchers. The Cochrane collaboration tool was utilized for risk bias assessment, and Revman 5.3 along with Stata 17.0 were employed for statistical analysis. Outcomes: We searched 254 literature, and 19 were included in this meta-analysis. The results showed that Dl-3-n-Butylphthalide improved the clinical efficacy rate (RR = 1.25, 95% CI 1.19-1.31; p = 0.00), quality of life (MD = 13.93, 95% CI: 11.88-15.98; p = 0.000), increased cerebral blood flow and velocity, reduced cerebral edema volume, Hcy concentration, and did not have obvious adverse reactions (RR = 0.68, 95% CI: 0.39-1.18; p = 0.10). Conclusion: This meta-analysis is the first to demonstrate the potential of Dl-3-n-Butylphthalide in treating cerebral hemorrhage. It suggests that Dl-3-n-Butylphthalide may alleviate clinical symptoms by modulating neurological function and improving hemodynamics. Our findings provide robust evidence for incorporating Dl-3-n-Butylphthalide into cerebral hemorrhage treatment strategies, potentially guiding future clinical practice and research. Systematic Review Registration: https://www.crd.york.ac.uk/PROSPERO/ display_record.php?RecordID=355114, Identifier CRD42022355114.

A gut-derived hormone regulates cholesterol metabolism.

The reciprocal coordination between cholesterol absorption in the intestine and de novo cholesterol synthesis in the liver is essential for maintaining cholesterol homeostasis, yet the mechanisms governing the opposing regulation of these processes remain poorly understood. Here, we identify a hormone, Cholesin, which is capable of inhibiting cholesterol synthesis in the liver, leading to a reduction in circulating cholesterol levels. Cholesin is encoded by a gene with a previously unknown function (C7orf50 in humans; 3110082I17Rik in mice). It is secreted from the intestine in response to cholesterol absorption and binds to GPR146, an orphan G-protein-coupled receptor, exerting antagonistic downstream effects by inhibiting PKA signaling and thereby suppressing SREBP2-controlled cholesterol synthesis in the liver. Therefore, our results demonstrate that the Cholesin-GPR146 axis mediates the inhibitory effect of intestinal cholesterol absorption on hepatic cholesterol synthesis. This discovered hormone, Cholesin, holds promise as an effective agent in combating hypercholesterolemia and atherosclerosis.

Exploration of 2D and 3D-QSAR analysis and docking studies for novel dihydropteridone derivatives as promising therapeutic agents targeting glioblastoma.

Background: Dihydropteridone derivatives represent a novel class of PLK1 inhibitors, exhibiting promising anticancer activity and potential as chemotherapeutic drugs for glioblastoma. Objective: The aim of this study is to develop 2D and 3D-QSAR models to validate the anticancer activity of dihydropteridone derivatives and identify optimal structural characteristics for the design of new therapeutic agents. Methods: The Heuristic method (HM) was employed to construct a 2D-linear QSAR model, while the gene expression programming (GEP) algorithm was utilized to develop a 2D-nonlinear QSAR model. Additionally, the CoMSIA approach was introduced to investigate the impact of drug structure on activity. A total of 200 novel anti-glioma dihydropteridone compounds were designed, and their activity levels were predicted using chemical descriptors and molecular field maps. The compounds with the highest activity were subjected to molecular docking to confirm their binding affinity. Results: Within the analytical purview, the coefficient of determination (R2) for the HM linear model is elucidated at 0.6682, accompanied by an R2 cv of 0.5669 and a residual sum of squares (S2) of 0.0199. The GEP nonlinear model delineates coefficients of determination for the training and validation sets at 0.79 and 0.76, respectively. Empirical modeling outcomes underscore the preeminence of the 3D-QSAR model, succeeded by the GEP nonlinear model, whilst the HM linear model manifested suboptimal efficacy. The 3D paradigm evinced an exemplary fit, characterized by formidable Q2 (0.628) and R2 (0.928) values, complemented by an impressive F-value (12.194) and a minimized standard error of estimate (SEE) at 0.160. The most significant molecular descriptor in the 2D model, which included six descriptors, was identified as "Min exchange energy for a C-N bond" (MECN). By combining the MECN descriptor with the hydrophobic field, suggestions for the creation of novel medications were generated. This led to the identification of compound 21E.153, a novel dihydropteridone derivative, which exhibited outstanding antitumor properties and docking capabilities. Conclusion: The development of 2D and 3D-QSAR models, along with the innovative integration of contour maps and molecular descriptors, offer novel concepts and techniques for the design of glioblastoma chemotherapeutic agents.

Combined Molecular and Morphological Imaging of CTCs for HER2-Targeted Chemotherapy Guidance.

Since biomolecules change dynamically with tumor evolution and drug treatment, it is necessary to confirm target molecule expression in real time for effective guidance of subsequent chemotherapy treatment. However, current methods to confirm target proteins require complex processing steps and invasive tissue biopsies, limiting their clinical utility for targeted treatment monitoring. Here, CTCs, as a promising liquid biopsy source, were used to molecularly characterize the target protein HER2. To accurately identify CTCs, we specifically proposed a combined molecular and morphological imaging method, rather than using specific biomarker alone or morphology analysis, we identified CTCs as CK19+/CD45-/HE+. On the basis of the accurate identification of CTCs, we further analyzed the target protein HER2 in clinical patients at the single-CTC level. Comparative analysis of the clinical results of patient pathological tissue and paired blood samples showed that CTCs had a heterogeneous HER2 expression at the single-cell level and showed results inconsistent with the immunohistochemistry results in some cases. CTC-based analysis could help clinicians have a more comprehensive understanding of patient target protein expression. We believe that CTC-based target protein studies are of great significance for the precise management of targeted therapy.

Effects of NBP injection on the inflammatory response, oxidative stress response and vascular endothelial function in patients with ACI: A systematic review and meta-analysis.

BACKGROUND: Acute cerebral infarction (ACI) is a common medical emergency. This study is the first systematic review of the use of Dl-3-n-butylphthalide (NBP) injection in the treatment of ACI. The purpose of this study was to systematically evaluate the effects of NBP injection on the inflammatory response, oxidative stress response and vascular endothelial function in patients with acute ACI. The objective is to provide reference for clinical application. METHODS: From the establishment of the database until August 2022, we systematically searched EMbase, PubMed, Cochrane Library, Web of Science, CNKI, VIP, and Wanfang Database. RCTs and retrospective studies were included in this study, and the results that qualified for inclusion were screened by 2 researchers and cross-checked. After the relevant data were extracted, a meta-analysis was performed using RevMan5.3 software. RESULTS: A total of 3307 patients with ACI from 34 studies were analyzed. The meta-analysis showed that the C-reactive protein levels in the NBP combined group were effectively reduced compared with those in the control group (MD = -3.75, 95% confidence intervals [95% CI] [-4.95, -2.56], P < .00001). Based on comparison with the control group, it is evident that combination treatment with NBP is more effective than control group in reducing the oxidative stress response of ACI (MD[superoxide dismutase levels] = 22.16, 95% CI [14.20,30.11], P < .00001; MD[malondialdehyde levels] = -1.97, 95% CI [-2.62, -1.32], P < .00001). Comparison with the control group shows that combination treatment with NBP is more effective in improving vascular endothelial function in ACI patients (MD[vascular endothelial growth factor levels] = 71.44, 95% CI [41.22, 101.66], P < .00001; MD[endothelin-1 levels] = -11.47, 95% CI [-17.39, -5.55], P = .0001; MD[nitric oxide levels] = 9.54, 95% CI [8.39, 10.68], P < .00001) than control group. The NBP combined group also showed a greater reduction in cerebral infarct volume (CIV) and cerebral infarct size (CIS) of ACI (MD[CIV] = -1.52, 95% CI [-2.23, -0.81], P < .0001; MD[CIS] = -2.79, 95% CI [-3.65, -1.94], P < .00001). The NBP combined group did not show an increase in the incidence of adverse reactions compared with the control group (odds ratio = 1.06, 95% CI [0.73, 1.53], P = .77). CONCLUSION: In summary, the use of NBP in combination with control group for ACI can reduce the degree of nerve damage, reduce inflammation and oxidative stress, improve vascular endothelial function, and reduce CIS and CIV in ACI patients, without increasing the incidence of clinical adverse events.

Famsin, a novel gut-secreted hormone, contributes to metabolic adaptations to fasting via binding to its receptor OLFR796.

The intestine is responsible for nutrient absorption and orchestrates metabolism in different organs during feeding, a process which is partly controlled by intestine-derived hormones. However, it is unclear whether the intestine plays an important role in metabolism during fasting. Here we have identified a novel hormone, famsin, which is secreted from the intestine and promotes metabolic adaptations to fasting. Mechanistically, famsin is shed from a single-pass transmembrane protein, Gm11437, during fasting and then binds OLFR796, an olfactory receptor, to activate intracellular calcium mobilization. This famsin-OLFR796 signaling axis promotes gluconeogenesis and ketogenesis for energy mobilization, and torpor for energy conservation during fasting. In addition, neutralization of famsin by an antibody improves blood glucose profiles in diabetic models, which identifies famsin as a potential therapeutic target for treating diabetes. Therefore, our results demonstrate that communication between the intestine and other organs by a famsin-OLFR796 signaling axis is critical for metabolic adaptations to fasting.

The Learning Curve of Unilateral Biportal Endoscopic (UBE) Spinal Surgery by CUSUM Analysis.

Objective: To assess the learning curve of the unilateral biportal endoscopic (UBE) technique for the treatment of single-level lumbar disc herniation by cumulative summation (CUSUM) method analysis. Methods: A retrospective analysis was conducted to assess 97 patients' general condition, operation time, complications, and curative effect of single segmental UBE surgery performed by a spinal surgeon in his early stage of this technique. The learning curve of operation time was studied using a CUSUM method, and the cut-off point of the learning curve was obtained. Results: The operation time was 30 - 241(97.9 ± 34.7) min. The visual analog scale score of lower limb pain decreased from 5.75 ± 0.81 before the operation to 0.39 ± 0.28 at the last follow-up (P < 0.05). The Oswestry disability index score decreased from 66.48 ± 4.43 before the operation to 14.57 ± 3.99 at the last follow-up (P < 0.05). The CUSUM assessment of operation time revealed the learning curve was the highest in 24 cases. In the learning stage (1-24 cases), the operation time was 120.3 ± 43.8 min. In the skilled stage (25-97 cases), the operation time was 90.5 ± 27.8 min. Conclusions: About 24 cases of single segmental UBE operation are needed to master the UBE technique.

Question Answering System Based on Knowledge Graph in Traditional Chinese Medicine Diagnosis and Treatment of Viral Hepatitis B.

This article uses the real medical records and web pages of Chinese medicine diagnosis and treatment of hepatitis B to extract structured medical knowledge, and obtains a total of 8,563 entities, 96,896 relationships, 32 entity types, and 40 relationship types. The structured data was stored in the Neo4j graph structure database, and a knowledge graph of Chinese medical diagnosis and treatment of hepatitis B was constructed. The knowledge map is used as a structured data source to provide high-quality knowledge information for the medical question and answer system based on hepatitis B disease. Applying the deep learning method to the question identification and knowledge response of the question answering system makes the hepatitis B medical intelligent question answering system has important research and application significance. The question-and-answer system takes aim at hepatitis B, a public health problem in the world and leverages the advantages of traditional Chinese medicine for diagnosis and treatment. It provides a reference for doctors' disease diagnosis, treatment, and patient self-care. Its value is important for the treatment of hepatitis B disease.

Comparison of efficacy and safety of complementary and alternative therapies for essential hypertension with anxiety or depression disorder.

BACKGROUND: Essential hypertension (EH) with anxiety or depression belongs to the category of psycho-cardiology. Hypertension is closely related to anxiety and depression. The adverse reactions of Western medicine are apparent and the compliance is poor. Supplementary and replacement therapies have accumulated rich experience in clinical practices, which can reduce side effects and improve clinical efficacy. This study intends to use the Bayesian network meta-analysis (NMA) analysis method for the first time to gather randomized controlled trials (RCTs) related to complementary and alternative therapies in the treatment of hypertension with anxiety or depression disorder and rank efficacy and safety, to provide a reference basis for the treatment of hypertension with anxiety or depression disorder. METHODS: All randomized controlled trials (RCTs) and registered and ongoing trials of Chinese and English databases, related to supplementation and replacement therapies of EH with anxiety or depression disorder, published from initial state to February 2021, will be collected in the form of computer retrieval. Two researchers will independently screen the literature, extracting data, assessing bias risk and assessing heterogeneity. We will use software WinBUGS 1.4.3 and Stata 16.0 for pairwise meta-analysis and NMA to comprehensively evaluate various interventions. The quality of evidence will be evaluated through the Grading of Recommendations Assessment, Development and Evaluation (GRADE). RESULTS: This NMA will comprehensively compare and rank the efficacy and safety of a series of complementary and alternative therapies in treating EH with anxiety or depression disorder. CONCLUSION: Supplementary and replacement therapies have accumulated rich experience of clinical practices in improving EH with anxiety or depression disorder. We expect that this NMA will guide practice and research by providing reliable evidence of evidence-based medicine for the treatment of EH with anxiety or depression disorder. PROTOCOL REGISTRATION NUMBER: INPLASY202120068.

Comparison of efficacy and safety of complementary and alternative therapies for coronary heart disease complicated with anxiety or depression disorder: A protocol for Bayesian network meta-analysis.

BACKGROUND: With the acceleration of the pace of life, the phenomenon of anxiety and depression in patients with coronary heart disease (CHD) is more and more common, and "psycho-cardiology" arises spontaneously. At present, the drug treatments of psycho-cardiology are difficult to achieve satisfactory results, and the side effects are obvious. Complementary and replacement therapies of CHD complicated with anxiety or depression disorder play an increasingly positive role, but there is a lack of comparison among different complementary and alternative therapies. In this study, Bayesian network meta-analysis (NMA) analysis method will be used for the first time to synthesize all the evidences of direct and indirect comparison among a variety of interventions, and rank their effectiveness and safety. METHODS: Two independent researchers will search from the beginning to January 2021 mainly including randomized controlled trials (RCTs) and closely related ongoing RCTs of complementary and alternative therapies for CHD complicated with anxiety or depression disorder. And then identify, select and extract the data. The primary outcome measures are frequency of acute attack angina, severity of angina pectoris; the changed score in the validated scales, which can assess severity of anxiety or depression. Secondary outcomes include total efficacy rate, electrocardiogram improvement, traditional Chinese medicine symptoms score, changes of dosage of nitroglycerin and adverse effects. Using softwares WinBUGS 1.4.3 and STATA 16.0 for pairwise meta-analysis and NMA to comprehensively evaluate various interventions. The quality of evidences will be evaluated through the Grading of Recommendations Assessment, Development and Evaluation. RESULTS: This NMA will comprehensively compare and rank the efficacy and safety of a series of complementary and alternative therapies in the treatment of CHD complicated with anxiety or depression disorder. CONCLUSION: Supplementary and replacement therapies play an essential role in improving CHD complicated with anxiety or depression disorder. We expect that the NMA will provide reliable evidences of evidence-based medicine for treatment of CHD complicated with anxiety or depression disorder. PROTOCOL REGISTRATION NUMBER: INPLASY202120046. ETHICAL APPROVAL: This review does not require ethical approval.

Recent advances in microfluidic technologies for circulating tumor cells: enrichment, single-cell analysis, and liquid biopsy for clinical applications.

Circulating tumor cells (CTCs) detach from primary or metastatic lesions and circulate in the peripheral blood, which is considered to be the cause of distant metastases. CTC analysis in the form of liquid biopsy, enumeration and molecular analysis provide significant clinical information for cancer diagnosis, prognosis and therapeutic strategies. Despite the great clinical value, CTC analysis has not yet entered routine clinical practice due to lack of efficient technologies to perform CTC isolation and single-cell analysis. Taking the rarity and inherent heterogeneity of CTCs into account, reliable methods for CTC isolation and detection are in urgent demand for obtaining valuable information on cancer metastasis and progression from CTCs. Microfluidic technology, featuring microfabricated structures, can precisely control fluids and cells at the micrometer scale, thus making itself a particularly suitable method for rare CTC manipulation. Besides the enrichment function, microfluidic chips can also realize the analysis function by integrating multiple detection technologies. In this review, we have summarized the recent progress in CTC isolation and detection using microfluidic technologies, with special attention to emerging direct enrichment and enumeration in vivo. Further, few insights into single CTC molecular analysis are also demonstrated. We have provided a review of potential clinical applications of CTCs, ranging from early screening and diagnosis, tumor progression and prognosis, treatment and resistance monitoring, to therapeutic evaluation. Through this review, we conclude that the clinical utility of CTCs will be expanded as the isolation and analysis techniques are constantly improving.

The tight junction protein TJP1 regulates the feeding-modulated hepatic circadian clock.

Circadian clocks in the suprachiasmatic nucleus and peripheral tissues orchestrate behavioral and physiological activities of mammals in response to environmental cues. In the liver, the circadian clock is also modulated by feeding. However, the molecular mechanisms involved are unclear. Here, we show that TJP1 (tight junction protein 1) functions as a mediator of mTOR (mechanistic target of rapamycin) to modulate the hepatic circadian clock. TJP1 interacts with PER1 (period circadian regulator 1) and prevents its nuclear translocation. During feeding, mTOR phosphorylates TJP1 and attenuates its association with PER1, thereby enhancing nuclear shuttling of PER1 to dampen circadian oscillation. Therefore, our results provide a previously uncharacterized mechanistic insight into how feeding modulates the hepatic circadian clock.

Phenotype-related drug sensitivity analysis of single CTCs for medicine evaluation.

Due to the heterogeneous and variable drug sensitivity of tumor cells, real-time monitoring of a patient's drug response is desirable for implementing personalized and dynamic therapy. Although considerable efforts have been directed at drug screening in living cells, performing repeated drug sensitivity analysis using patient-derived primary tumor cells at the single-cell level remains challenging. Here, we present an efficient approach to assess phenotype-related drug sensitivity at the single-cell level using patient-derived circulating tumor cells (CTCs) based on a drug sensitivity microfluidic chip (DS-Chip). The DS-Chip consists of a drug gradient generator and parallel cell traps, achieving continuous single CTC capture, drug gradient distributions, drug stimulation, fluorescent probe labeling and three-color fluorescence imaging. Based on the established DS-Chip, we investigated the drug sensitivity of single cells by simultaneously monitoring epithelial-mesenchymal transition (EMT) biomarkers and apoptosis in living cells, and verified the correlation between EMT gradients and drug sensitivity. Using the new approach, we further tested the optimal drug response dose in individual CTCs isolated from 5 cancer patients through fluorescence analysis of EMT and apoptosis. The DS-Chip allows noninvasive and real-time measurements of the drug sensitivity of a patient's tumor cells during therapy. This developed approach has practical significance and can effectively guide drug selection and therapeutic evaluation for personalized medicine.

1-Deoxynojirimycin modulates glucose homeostasis by regulating the combination of IR-GlUT4 and ADIPO-GLUT4 pathways in 3T3-L1 adipocytes.

1-Deoxynojirimycin (DNJ), the main alkaloid in mulberry leaves, was recognized to treat patients with type 2 diabetes mellitus (T2DM). However, the regulatory mechanism of DNJ on glucose homeostasis was still unclear. In the present study, a safe concentration of 0.1-10 µmol/L for DNJ was incubated with mature 3T3-L1 adipocytes. The results demonstrated that the genes/proteins expression of insulin receptor (IR), phosphatidylinositol-3-kinase (PI3K), protein kinase B (AKt/PkB), and adiponectin (ADIPO) increased with the increasing of DNJ concentration from 0.1-10 µmol/L. However the mRNA expression of adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK), glucose transporter 4 (GLUT4) and glucose absorption increased to the maximum at concentration of 5 µmol/L then decreased with further increase of DNJ concentration to 10 µmol/L. Both IR and ADIPO signaling pathways simultaneously affect the glucose homeostasis regulation effect of DNJ, whereas the key response target located in AMPK and its effect on subsequent GLUT4 mRNA expression.

Regulation of glucose and lipid metabolism in health and disease.

Glucose and fatty acids are the major sources of energy for human body. Cholesterol, the most abundant sterol in mammals, is a key component of cell membranes although it does not generate ATP. The metabolisms of glucose, fatty acids and cholesterol are often intertwined and regulated. For example, glucose can be converted to fatty acids and cholesterol through de novo lipid biosynthesis pathways. Excessive lipids are secreted in lipoproteins or stored in lipid droplets. The metabolites of glucose and lipids are dynamically transported intercellularly and intracellularly, and then converted to other molecules in specific compartments. The disorders of glucose and lipid metabolism result in severe diseases including cardiovascular disease, diabetes and fatty liver. This review summarizes the major metabolic aspects of glucose and lipid, and their regulations in the context of physiology and diseases.

Consecutive Sorting and Phenotypic Counting of CTCs by an Optofluidic Flow Cytometer.

Circulating tumor cell (CTC) analysis has been approved for cancer diagnosis and monitoring. However, efficient sorting and high-through phenotypic counting of CTCs from peripheral blood is still a challenge. In this manuscript, we propose an optofluidic flow cytometer (OFCM), which integrates a multistage microfluidic chip and a four-color fluorescence detection system. The OFCM can automatically complete CTC separation, 3D focusing in the microchannel, single-cell phenotypic analysis, and counting at 1.2 mL of whole blood/hour. A high recovery greater than 95% was obtained. Using the OFCM, we analyzed the epithelial-to-mesenchymal transition (EMT) phenotype of CTCs in patients with breast cancer and patients with nonsmall cell lung cancer, which proved that the OFCM is adaptable for phenotypic counting of various CTCs based on the fluorescence labeling of varied biomarkers. We believe that this OFCM will provide a convenient and efficient device for clinical liquid biopsy of tumors.