GSZ Formula Enhances Sleep Quality:Exploring Its Active Ingredients and Mechanism Using a Network Medicine Approach

Background:Sleep is essential for maintaining human health,and insomnia is a widespread problem.Traditional Chinese medicine(TCM)has been used for centuries to treat sleep disorders,with fewer reported side effects compared to conventional treatments.Objective:This study seeks to investigate the sleep-promoting effects of the GSZ formula,which comprisesγ-aminobutyric acid(GABA),Schisandrae Chinensis Fructus(Wuweizi in Chinese),and Ziziphi Spinosae Semen(Suanzaoren in Chinese).In addition,this study aims to explore the active ingredients and potential mechanisms underlying the sleep-enhancing effects of the formula.Methods:The impact of GSZ on sleep was evaluated using two models,the complete sleep model and the sub-threshold sleep model.Mice were randomly divided into five groups and orally administered GSZ solution(0.33 g/kg/day or 0.99 g/kg/day),positive drug diazepam(2.50 mg/kg)or a control solution for 30 days.Hypno-sis model was established in mice using pentobarbital sodium.Sleep duration and incidence were measured by recording when the righting reflex of mice disappeared for more than 1 min.GABA and dopamine(DA)levels in mouse brain tissue were measured using ELISA kits.The ingredients of the GSZ formula were identified using mass spectrometry,and the targets of these ingredients and disease-related genes were retrieved from public databases.A network medicine approach was used to calculate the shortest path between ingredient targets and disease-related proteins.The expression levels of potential proteins,such as Akt,p-Akt,GSK-3β,and p-GSK-3β,were analyzed using Western blotting based on the predicted results.Results:GSZ significantly prolonged sleep duration and enhanced the sleep rate in mice(P<0.05).Furthermore,it elevated GABA levels and reduced DA levels in the mouse brain(P<0.05).Network medicine analysis suggested that GABA,stearic acid,genistin,and coumestrol may be the most crucial active ingredients for sleep improve-ment.Western blotting analysis demonstrated that GSZ modulated the protein expression levels of p-Akt/Akt and p-GSK-3β/GSK-3β(P<0.05).Conclusion:Our study demonstrated that the GSZ formula could improve sleep,with key ingredients likely being GABA,stearic acid,genistin,and coumestrol.The mechanism might involve the regulation of the Akt/GSK-3βpathway,as revealed by the network medicine analysis and experimental validation.Our current new findings shed light on the potential mechanisms underlying the sleep-enhancing effects of the GSZ formula,which could provide experimental evidence to develop innovative treatments for insomnia.

Network Curriculum Design of Traditional Chinese Medicine Pharmacy Specialty Formed by Introducing Guangxi Minority Medicine

Starting from the background and significance of the development and design of the network course of traditional Chinese medicine pharmacy specialty formed by introducing Guangxi minority medicine,this paper discusses the teaching design scheme and module construction of the network course in the development process,and summarizes the design,production and construction experience of the network course.Through practice,the teaching effect and achievement of the network course of traditional Chinese medicine pharmacy specialty formed by introducing Guangxi minority medicine are obvious.

MicroRNA–disease Network Analysis Repurposes Methotrexate for the Treatment of Abdominal Aortic Aneurysm in Mice

Abdominal aortic aneurysm(AAA)is a permanent dilatation of the abdominal aorta and is highly lethal.The main purpose of the current study is to search for noninvasive medical therapies for AAA,for which there is currently no effective drug therapy.Network medicine represents a cuttingedge technology,as analysis and modeling of disease networks can provide critical clues regarding the etiology of specific diseases and therapeutics that may be effective.Here,we proposed a novel algorithm to quantify disease relations based on a large accumulated microRNA–disease association dataset and then built a disease network covering 15 disease classes and 304 diseases.Analysis revealed some patterns for these diseases.For instance,diseases tended to be clustered and coherent in the network.Surprisingly,we found that AAA showed the strongest similarity with rheumatoid arthritis and systemic lupus erythematosus,both of which are autoimmune diseases,suggesting that AAA could be one type of autoimmune diseases in etiology.Based on this observation,we further hypothesized that drugs for autoimmune diseases could be repurposed for the prevention and therapy of AAA.Finally,animal experiments confirmed that methotrexate,a drug for autoimmune diseases,was able to alleviate the formation and development of AAA.

Exploring Traditional Chinese Medicine by A Novel Therapeutic Concept of Network Target

Traditional Chinese medicine（TCM） holds a holistic theory, and specializes in balancing disordered human body using numerous natural products, particularly Chinese herbal formulae. TCM has certain treatment advantages for patients suffering from various complex diseases. However, due to the complex nature of TCM, it remains difficult to unveil such holistic medicine by the current reductionism research strategies, which treat both herbal ingredients and targets in isolation. Recently, an emerging network pharmacology approach has been introduced to tackle this bottleneck problem. A TCM-derived novel therapeutic concept, ＂network target＂, which is different from the Western medicine＇s ＂onetarget＂ concept, has been proposed from China. The network target strategy is able to illustrate the complex interactions among the biological systems, drugs, and complex diseases from a network perspective, and thus provides an innovative approach to access ancient remedies in a precision manner and at a systematic level, which also highlights TCM＇s potential in current medical systems.

Efficacy of Traditional Chinese Medicines in Preventing Oxaliplatin-induced Peripheral Neurotoxicity in Cancer Patients:A Network Meta-analysis

Objective Oxaliplatin-induced peripheral neurotoxicity（OIPN） is the main limitation for its continuation in cancer patients.Traditional Chinese medicines（TCMs） have been used to prevent OIPN in China and have been demonstrated to be effective.However,due to the lack of direct comparison among TCMs,it remains unclear that which TCM is the best for OIPN prevention.Consequently,the present study aimed to compare the relative efficacies of TCMs to find out the best TCM by applying a network meta-analysis.Methods Studies were identified by searching PubMed,EMbase,Cochrane Libraries,CNKI,WanFang,and WeiPu database from January 1990 to May2016.Randomized controlled trials（RCTs） that evaluated the efficacy of TCMs in preventing OIPN in cancer patients were included.Statistical analysis was performed with ADDIS 1.1 6.6.Results Twenty-five RCTs（1572 patients） involving five TCMs were included.The results of network meta-analyses showed that compared with oxaliplatin-based chemotherapy alone,the combination with Huangqi Injection（HQI）,Shenmai Injection（SMI）,Shenfu Injection（SFI）,Buyang Huanwu Decoction（BHD）,and Huangqi Guizhi Wuwu Decoction（HGWD） could decrease the overall OIPN incidence and the severe OIPN incidence in cancer patients.In addition,probability ranking results showed the order of efficacy in preventing overall OIPN incidence was HQI 〉 HGWD 〉 SFI =SMI 〉 BHD,while the order of efficacy in preventing severe OIPN incidence was HQI 〉HGWD 〉 BHD 〉 SFI = SMI.Conclusion All five TCMs are effective neuroprotective agents against OIPN.Among these TCMs,HQI and HGWD were superior to others in clinical efficacy.Moreover,Astragalus membranaceus might be a more promising herb for the OIPN prevention.However,more direct head-to-head RCTs with high quality and large sample size are still needed to further confirm the conclusion.

Integrated Genomic and Network-Based Analyses of Complex Diseases and Human Disease Network

A disease phenotype generally reflects various pathobiological processes that interact in a complex network. The highly interconnected nature of the human protein interaction network（interactome） indicates that, at the molecular level, it is difficult to consider diseases as being independent of one another. Recently, genome-wide molecular measurements, data mining and bioinformatics approaches have provided the means to explore human diseases from a molecular basis. The exploration of diseases and a system of disease relationships based on the integration of genome-wide molecular data with the human interactome could offer a powerful perspective for understanding the molecular architecture of diseases. Recently, subnetwork markers have proven to be more robust and reliable than individual biomarker genes selected based on gene expression profiles alone, and achieve higher accuracy in disease classification. We have applied one of these methodologies to idiopathic dilated cardiomyopathy（IDCM） data that we have generated using a microarray and identified significant subnetworks associated with the disease. In this paper, we review the recent endeavours in this direction, and summarize the existing methodologies and computational tools for network-based analysis of complex diseases and molecular relationships among apparently different disorders and human disease network. We also discuss the future research trends and topics of this promising field.

Brain functional connectivity network studies of acupuncture： a systematic review on resting-state fMRI

Background： Functional magnetic resonance imaging （fMRI） is a novel method for studying the changes of brain networks due to acupuncture treatment. In recent years, more and more studies have focused on the brain functional connectivity network of acupuncture stimulation. Objective： To offer an overview of the different influences of acupuncture on the brain functional connec- tivity network from studies using resting-state fMRI. Search strategy： The authors performed a systematic search according to PRISMA guidelines, The database PubMed was searched from January 1, 2006 to December 31, 2016 with restriction to human studies in English language. Inclusion criteria： Electronic searches were conducted in PubMed using the keywords ＂acupuncture＂ and ＂neuroimaging＂ or ＂resting-state fMRI＂ or ＂functional connectivity＂, Data extraction and analysis： Selection of included articles, data extraction and methodological quality assessments were respectively conducted by two review authors. Results： Forty-four resting-state fMRI studies were included in this systematic review according to inclu- sion criteria. Thirteen studies applied manual acupuncture vs. sham, four studies applied electro- acupuncture vs. sham, two studies also compared transcutaneous electrical acupoint stimulation vs. sham, and nine applied sham acupoint as control. Nineteen studies with a total number of 574 healthy subjects selected to perform fMRI only considered healthy adult volunteers. The brain functional connec- tivity of the patients had varying degrees of change. Compared with sham acupuncture, verum acupunc- ture could increase default mode network and sensorimotor network connectivity with pain-, affective- and memory-related brain areas. It has significantly greater connectivity of genuine acupuncture between the periaqueductal gray, anterior cingulate cortex, left posterior cingulate cortex, right anterior insula, limbic/paralimbic and precuneus compared with sham acupuncture. Some research had also shown that acupuncture could adjust the limbic-paralimbic-neocortical network, brainstem, cerebellum, subcortical and hippocampus brain areas. Conclusion： It can be presumed that the functional connectivity network is closely related to the mech- anism of acupuncture, and central integration plays a critical role in the acupuncture mechanism.

Identification of Hypertension Subgroups through Topological Analysis of Symptom-Based Patient Similarity

Objective:To obtain the subtypes of the clinical hypertension population based on symptoms and to explore the relationship between hypertension and comorbidities.Methods:The data set was collected from the Chinese medicine(CM)electronic medical records of 33,458 hypertension inpatients in the Affiliated Hospital of Shandong University of Traditional Chinese Medicine between July 2014 and May 2017.Then,a hypertension disease comorbidity network(HDCN)was built to investigate the complicated associations between hypertension and their comorbidities.Moreover,a hypertension patient similarity network(HPSN)was constructed with patients'shared symptoms,and 7 main hypertension patient subgroups were identified from HPSN with a community detection method to exhibit the characteristics of clinical phenotypes and molecular mechanisms.In addition,the significant symptoms,diseases,CM syndromes and pathways of each main patient subgroup were obtained by enrichment analysis.Results:The significant symptoms and diseases of these patient subgroups were associated with different damaged target organs of hypertension.Additionally,the specific phenotypic features(symptoms,diseases,and CM syndromes)were consistent with specific molecular features(pathways)in the same patient subgroup.Conclusion:The utility and comprehensiveness of disease classification based on community detection of patient networks using shared CM symptom phenotypes showed the importance of hypertension patient subgroups.

Anti-Inflammatory Effect of Qingwen Baidu Decoction (清瘟败毒饮) in Sepsis Rats

Objective：To explore the pharmacological anti-inflammatory mechanism of Chinese formula Qingwen Baidu Decoction（清瘟败毒饮,QBD） from the view of holistic biology.Methods：The rats were randomly divided into a normal conrol group,a lipopolysaccharide（LPS） group,the low- and high-dose QBD groups,and a dexamethasone（DXM） group.NR8383 cells were treated with culture fluid containing 6%serum from rats of each group respectively.Inflammatory mediators were detected by reverse transcription polymerase chain reaction（RT-PCR）,Western blotting hybridization,enzyme linked immunosorbent assay（ELISA）,polymerase chain reaction（PCR） gene array and antibody array.Results：It is showed that the levels of interleukin（IL）-1 α,IL-4 and IL-12 were enhanced in the low-dose QBD group;levels of IL-1 α,IL-12 and IL-18 were augmented in the high-dose QBD group,compared with the LPS group after ELISA detection.Western blot showed that IL-1β and tumor necrosis factor（TNF）-α expression of the control group were lower than other groups.IL-1 βlevel of the low-dose and high-dose QBD groups detected by RT-PCR was higher in early stage but lower after24 h than that of the control group（P〈0.01）.Expression of 84 main inflammatory cytokines and receptors was detected by rat inflammatory cytokines and receptors PCR array.Up-regulation genes were 22 in both the LPS group and the low-dose QBD group,among which 16 up-regulating genes were the same.In these 16 genes,the up-regulating amplitude of 9 genes in the low-dose QBD group was less than that in the LPS group,4 were similar to and 3 were more.Twenty-nine main cytokines were inspected by rat cytokine antibody array.Intergroup gray value differences were found in 7 expressed cytokines.The levels of these 7 cytokines in the lowdose QBD group were all lower than those in the the LPS group.Conclusions：QBD has anti-inflammatory effect on sepsis by changing the level of inflammatory mediators.