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@#【Objective】 To study the composition and function of tongue coating (TC) and gastrointestinal tract (GIT) microbiota in participants with yellow-greasy tongue coating (YGTC), and to explore the representative metabolite markers and pathways in this group. 【Methods】 Subjects with YGTC or thin-white tongue coating (TWTC) were recruited from December 1, 2021 to October 30, 2022, and the TC and fecal samples were collected. Samples were subjected to both whole-genome shotgun (WGS), and 16S rRNA gene sequencing. The α-diversity analysis, principal component analysis (PCA), and Spearman correlation analysis were performed for two groups. Ultra-performance liquid chromatography combined with tandem mass spectrometry (UPLC–MS/MS) analysis was used to analyze metabolomics and enrichment of metabolic pathways. 【Results】 The results revealed 20 YGTC participates and 19 TWTC participates. At the genus level, the dominant bacterial species of TC flora and intestinal flora in the two groups were roughly the same, but the relative kurtosis difference was marked, and the abundance of potentially pathogenic bacteria in TC and fecal samples of YGTC subjects was higher. There were 9 down-regulated microorganisms in the TC samples, 26 down-regulated microorganisms, and 6 up-regulated microorganisms in YGTC subjects. The α-diversity analysis indicated that the Chao and abundance-based coverage estimator (ACE) indices of TC bacteria in the YGTC subjects showed a decreasing trend, but the difference was not statistically significant (P > 0.05). The α-diversity of fecal samples and the Chao and ACE indices decreased significantly (P < 0.05). PCA showed that the microflora structure of TC and fecal samples were significantly different between the two groups. Spearman correlation analysis showed that there was no correlation between TC and fecal microorganisms at phyla and genus levels in the same subjects (P > 0.05). The metabolomics results demonstrated that fumarate reductase, V/A ATPase, and phosphatidylethanolamine were increased, and glycerate-3p, UDP-glucose, and quinone oxidoreductase metabolites were decreased in YGTC TC samples. Inosine monophosphate (IMP), uridine monophosphate (UMP), and gamma-aminobutyric acid(GABA) were increased in YGTC fecal samples, while the contents of ribo-5P, histidine, biotin,and cobalamin were decreased. Metabolic pathway analysis indicated that the abundance of the TC and fecal samples of the YGTC subjects was relatively low in various metabolic pathways, including amino acid metabolism, carbohydrate metabolism, nitrogen metabolism, and energy metabolism. 【Conclusion】 Structural and functional changes in TC and GIT microbiota or metabolite markers could be potential biological bases of YGTC formation.
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@#【Objective】 To explore the development and research hotspots on the application of artificial intelligence (AI) in traditional Chinese medicine (TCM) diagnosis and predict research trends in the area. 【Methods】 All articles were retrieved from China National Knowledge Infrastructure (CNKI), Wanfang Data (Wanfang), China Science and Technology Journal Database (VIP), and Web of Science Core Collection (WoSCC). All related papers published in journals from the foundation of the databases to December 31, 2022 were included. NoteExpress, Co-Occurrence(COOC), VOSviewer, and CiteSpace were used to visualize data about publication volumes, journals, authors, research institutions, and keywords as well as to analyze hotspots trending topics in the field. 【Results】 A total of 686 articles were retrieved from the databases, among which 610 papers were published in Chinese and 76 in English. In terms of the journals in which these papers were published, 238 of them were Chinese journals and 52 were English ones. The number of the papers published in journals presented a slow growth. According to the results from Chinese article analysis, WANG Yiqin from Shanghai University of Traditional Chinese Medicine published the most papers in the field. The authors of Chinese papers belonged to six long-term research teams, led by WANG Yiqin and XU Jiatuo (Shanghai University of Traditional Chinese Medicine), WEI Yuke (Guangdong University of Technology), LI Gang (Tianjin University), XI Guangcheng (Institute of Automation of the Chinese Academ of Sciences), and NIU Xin (Beijing University of Chinese Medicine), respectively. In accordance with results from English paper analysis, four authors equally publishing the most papers were YAN Haixia, HU Xiaojuan, and JIANG Tao (Shanghai University of Traditional Chinese Medicine), and WEN Chuanbiao (Chengdu University of Traditional Chinese Medicine). The authors of English papers were from two major research teams in the field of Shanghai University of Traditional Chinese Medicine. Currently, research hotspots on AI such as neural networks, data mining, machine learning, feature recognition, image processing, and expert systems, have been centered on tongue diagnosis, pulse diagnosis, and syndrome research in TCM. Additionally, it was found that research on the topic was gradually evolving from explorations of a single diagnosis method to investigations on the combination of multiple TCM diagnosis methods. 【Conclusion】 Research on AI application in TCM diagnosis is still in a slowly growing stage. As technology develops, AI has been applied to many aspects of TCM diagnosis. Therefore, how to combine the two for improving TCM diagnosis is something worthy of our brainstorming and exploring.
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Objective@#To explore the differential expression profiles of DNA methylation sites/regions and potential molecular mechanisms in the peripheral blood of coronary heart disease (CHD)-induced unstable angina pectoris patients with or without Qi deficiency and blood stasis syndrome, and to provide scientific evidence for the conbination of disease and syndrome.@*Methods@#According to the pre-determined inclusion and exclusion criteria, the study subjects were enrolled and divided into two groups namely CHD-induced unstable angina group (G group) and healthy control group (J group) to conduct “disease” analysis, while G group was further divided into Qi deficiency and blood stasis syndrome group (case group) and non-Qi deficiency blood stasis syndrome group (control group) to perform “syndrome” analysis. The general data and clinical information of the study subjects were collected. The peripheral venous blood was extracted on an empty stomach, and the Illumina Infinium MethylationEPIC BeadChip (850K methylation chip) was used to detect the differential expressionprofiles of DNA methylation in each group, ChAMP software (V 2.14.0) was used for the differential methylation data analysis, with a threshold of the adjusted P value (adj.P.val) < 0.01. Gene Ontology (GO) and Kyoto Encyclopedia of Genomes (KEGG) were employed for the functional and pathway enrichment analyses of related mapped genes.@*Results@#A total of 263 differentially methylated CpG positions (DMPs) were screened out between G and J groups, including 191 hypermethylated positions such as cg05845204 and cg08906898, and 72 hypomethylated positions such as cg26919182 and cg13149459. These positions were mainly mapped to 148 genes encompassing RNA binding motif protein 39 (RBM39), acetyl-CoA acyltransferase 2 (ACAA2), protein phosphatase 1 regulatory subunit 12B (PPP1R12B), and the dual-specificity tyrosine phosphorylation-regulated kinase 2 (DYRK2). GO functional enrichment analysis revealed that the genes of the DMPs were primarily enriched in protein localization to chromosomes, regulation of cell morphogenesis, negative regulation of calcium-mediated signals, etc. KEGG pathway analysis suggested that the genes were mainly enriched in fatty acid metabolism and endocytosis pathways. In addition, a total of 23 differential methylation regions (DMRs) were identified, with overlapping genes such as transmembrane protein 232 (TMEM232), ribosomal protein large P1 (RPLP1), peroxisomal biogenesis factor 10 (PEX10), and forkhead box N3 (FOXN3) recognized. It was found that GO functions were mainly enriched in the negative regulation of Ras protein signal transduction, small GTPase-mediated signal transduction, negative regulation, etc. A total of 1 703 differential methylation sites were screened out between case and control groups, including 444 increased methylation positions such as cg05573767 and 1 259 decreased methylationpositions such as cg19938535, and cg03893872. These positions were mapped to 1 108 genes such as ribosomal protein S6 kinase A2 (RPS6KA2), leucine rich repeat containing 16A (LRRC16A), and hedgehog acyltransferase (HHAT). According to the GO functional enrichment analysis, the genes relating to the DMPs were mainly enriched in biological functions such as transmembrane receptor protein serine/threonine kinase signaling pathway and axonogenesis. The KEGG pathway enrichment analysis suggested the involvement of Rap1 signaling pathway, adenosine 5’-monophosphate-activated protein kinase (AMPK) signaling pathway, etc. A total of 21 DMRs were identified, including 22 overlapping genes such as mucin 4 (MUC4), three prime repair exonuclease 1 (TREX1), and LIM homeobox 6 (LHX6). GO analysis demonstrated that the genes primarily participated in molecular functions such as positive regulation of transmembrane transport, regulation of fatty acid metabolism, and copper ion binding.@*Conclusion@#This study reveals the methylation patterns of DMPs and DMRs in patients with Qi deficiency and blood stasis syndrome caused by CHD-induced unstable angina pectoris. Potential epigenetic regulation of fatty acid metabolism, Rap1 signaling, and other molecular functions are involved in the development of CHD between the "disease" and "syndrome".
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@#Objective This study examined the research status and development process of FU Qingzhu’s Obstetrics and Gynecology (Fu Qing Zhu Nv Ke,《傅青主女科》, FQZNK) in the past 40 years with bibliometrics and visual analysis. Methods Retrieved all related literature in the research field of FQZNK from the domestic and foreign databases: China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (VIP), Wanfang Database, and Web of Science (WOS) core database, including Science Citation Index Expanded (SCIE), Social Sciences Citation Index (SSCI), and Arts & Humanities Citation Index (A&HCI). The search range was from January 1, 1980 to March 10, 2021. In addition, bibliometrics and CiteSpace 5.7.R2 software were used to analyze literature types, published journals, cited literature, the number of author publications, co-author networks, co-institution networks, keyword co-occurrence networks, keyword clusters, and keyword bursts. Results A total of 678 valid records were included in the final dataset. Literature types, high publication journals, highly cited literature, high-yield institutions, high-yield research teams, and high-productivity scholars in this research field were found through bibliometrics. Literature types can be divided into four categories, among which 451 are theoretical studies on academic thoughts of FQZNK, accounting for 66.5% of the included journals. The Journal of Shanxi Traditional Chinese Medicine had the largest volume of published articles (61), accounting for 9.0% of the total number of the included journals. The most cited literature was ZHOU Mingxin’s article “Using the quantitative method to discuss author’s authenticity and formula characteristics of FU Qingzhu’s Obstetrics and Gynecology”, which was cited 94 times. Hunan University of Chinese Medicine, the institution with the most publications, published 45 articles, and YOU Zhaoling, the most published author, published 33 articles. Moreover, it was found that most high-yield researchers came from high-yield institutions and that Hunan University of Chinese Medicine had the most research on FQZNK. Keyword co-occurrence analysis revealed that the keyword “FQZNK” had the highest frequency (597 times) and the highest centrality (1.00). Keyword cluster analysis used the Log-Likelihood Ratio (LLR) algorithm to form eleven important clusters: #0 treatment aiming at its root causes, #1 gynecopathy, #2 Siwu Decoction (四物汤), #3 FU Qingzhu, #4 post-partum, #5 infertility, #6 dysmenorrhea, #7 sterility, #8 coordinate the heart and kidney, #9 Danggui Buxue Decoction (当归补血汤), and #10 treatment. It was found that the prescriptions of FQZNK were studied mainly before 2000, the theoretical studies were mainly conducted before 2010, and its clinical application was mainly explored from 2010 until now. Diseases such as dysmenorrhea, morbid vaginal discharge, infertility, metrorrhagia, and polycystic ovarian syndrome (PCOS) have recently become popular topics in this field. Conclusion The current study provides more scientific, accurate, and comprehensive scientific support for further research and development of traditional Chinese medicine (TCM) in FQZNK. With this foundation, people can use burst detection to ascertain the current hotspots in research, get their development trends, and forecast future research directions. In addition, infertility, morbid vaginal discharge, flooding, and PCOS treatments based on TCM syndrome differentiation are currently popular research topics for FQZNK.
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@#Objective To explore the effect and mechanism of Chaihu Longgu Muli Decoction (柴胡龙骨牡蛎汤, CHLGMLD) in rats with temporal lobe epilepsy (TLE). Methods A total of 80 Sprague-Dawley (SD) male rats were randomized into control (CON), model (MOD), carbamazepine (CBZ, 0.1 g/kg), CHLGMLD low dose (CHLGMLD-L, 12.5 g/kg), and high dose (CHLGMLD-H, 25 g/kg) groups, with 16 rats in each group. TLE rat models were established in the four groups with the use of lithium-pilocarpine except for the CON group. After the successful establishment of TLE models, all drugs were administered through gavage, and distilled water was given to rats in the CON and MOD groups for four weeks. The frequency and duration of seizures before and after treatment were recorded for the evaluation of the alleviation degree. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-146a-3p and miR-146a-5p. The expression levels of toll-like receptor 4 (TLR4), interleukin-1 receptor-associated kinase 1 (IRAK1), tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6), TAK1-binding protein (TAB), nuclear factor-kappa B (NF-κB), and interleukin-1 beta (IL-1β) in hippocampus were tested by immunofluorescence assay. Correlation analysis between the above factors and expressions of miR-146a-3p and miR-146a-5p were performed separately. Results CHLGMLD decreased the frequency (P < 0.05) and duration (P < 0.01) of seizures in rats. CHLGMLD down-regulated the expression levels of miR-146a-5p and miR-146a-3p (P < 0.05), and inhibited the expression levels of TLR4, IRAK1, TRAF6, TAB, NF-κB, and IL-1β (P < 0.01). The correlation analysis revealed that the expression levels of TLR4, IRAK1, TRAF6, TAB, NF-κB, and IL-1β were positively correlated with the expression levels of miR-146a-3p and miR-146a-5p detected by qRT-PCR, respectively (P < 0.01). Conclusion CHLGMLD can inhibite the TLR4 signaling pathway by lowering the expression levels of miR-146a-3p and miR-146a-5p to alleviate hippocampal dentate gyrus inflammation in TLE rats, thus relieving seizures.
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@#Objective To investigate the effect and underlying mechanism of Qingguang’an Granules (青光安颗粒剂, QGAG) on mitochondrial autophagy (mitophagy) of retinal ganglion cells (RGCs) in rats with chronic ocular hypertension (COH). Methods Sixty Sprague Dawley (SD) rats, half males and half females, were randomly assigned to three groups: the control, model, and QGAG (2.5 g/kg) groups, with 20 rats in each group. Rats’ model of COH was established by cauterizing episcleral veins in the model group and QGAG group. Three weeks after successful modeling, rats in the QGAG group were intragastrically administered with QGAG, while rats in the control group and the model group received an equal dose of normal saline. After three months of intragastric administration, intraocular pressure (IOP) of all rats was measured. The mitophagy was monitored by the immunofluorescence method, the mitochondrial membrane potential was measured using the JC-1 method, and the morphological changes of mitophagy in RGCs were observed by transmission electron microscopy. Meanwhile, rat RGCs were labeled using the fluorescent gold method, and RGCs density in each group was calculated. Moreover, RGCs apoptosis was observed by TdT-mediated dUTP Nick-End Labeling (TUNEL) assay. Finally, the expression levels of Parkin, optineurin, microtubule-associated protein 1 light chain 3-II/microtubule-associated protein 1 light chain 3-I (LC3-II/LC3-I), recombinant lysosomal associated membrane protein 1 (LAMP1), and B-cell lymphoma-2 (Bcl-2) in RGCs were determined by Western blot assay. The corresponding mRNAs were detected through quantitative real-time polymerase chain reaction (qRT-PCR). Results The QGAG reduced IOP in COH rats, and inhibited mitophagy and apoptosis of RGCs (P < 0.05). Besides, the QGAG significantly increased the expression levels of Parkin and Bcl-2 (P < 0.05), and inhibited the expression levels of optineurin, LAMP1, and LC3-II/LC3-I (P < 0.05) in RGCs of COH rats. Conclusion The QGAG can inhibit mitophagy in RGCs of COH rats and show a protective effect against optic nerve damage caused by glaucoma, which may be mediated through the mitophagy ubiquitination via the Parkin/PINK1-related pathway.