Clinical observations indicated a higher rate of obesity among children who received antibiotics at early ages. Experimental studies supported the role of the modified gut microbiome in the development of obesity as well. For identifying antibiotic classes that might promote or inhibit obesity-related dysbiosis, a database of the average yearly antibiotic consumption (2008–2018) has been developed using the European Center for Disease Prevention and Control (ECDC) yearly reports of antibiotic consumption in the community for the major antibiotic classes in 30 European countries, which were compared to the childhood and adult obesity prevalence featured in the Obesity Atlas. Pearson’s chi-square test was applied to estimate positive/negative correlations between antibiotic consumption and obesity. One-way ANOVA has been applied to test the differences in antibiotic consumption between groups, and logistic regression analysis was performed to determine the odds ratios (OR) of antibiotic consumption for obesity. Strong, positive associations were estimated between childhood obesity and the total consumption of systemic antibiotics, broad-spectrum, beta-lactamase-resistant penicillin, cephalosporin, and quinolone, and a negative correlation was found with the consumption of tetracycline, broad-spectrum, beta-lactamase-sensitive penicillin, and narrow-spectrum, beta-lactamase-sensitive penicillin. Our observation indicated that the “growth-promoting effect” of the consumption of certain antibiotic classes might be identified as a possible etiology in the development of obesity and might be the explanation for the obesity “pandemic”. Body mass index (BMI) is a simple weight-for-height index commonly used to classify overweightness and obesity (kg/m2). According to the WHO definition, overweightness is considered when the BMI is greater than or equal to 25, and obesity is a BMI greater than or equal to 30. For children aged between 5–19 years, overweightness is considered when the BMI-for-age is greater than one standard deviation above the WHO Growth Reference median, and obesity is estimated when the BMI is greater than two standard deviations above the WHO Growth Reference median .The human race has struggled to overcome food scarcity, disease, and hostile environments for centuries, but after the industrial revolution and the improving living conditions with the availability of nutrients, this situation changed. The recognition of obesity as a disease was theoretically established in 1948 by the World Health Organization (WHO), but for several years, medical professionals had ignored its importance  initiateObesity is a complex health issue resulting from a combination of causes and individual factors such as behavior and genetics. The most common causes of overweightness and obesity are well-known: genetics, the lack of physical activity, dietary patterns , medication use (steroids), certain diseases (hypothyreosis), etc. .The accidental discovery of the growth-promoting effect of antibiotics  soon folIt is suspected that the modification of the gut microbiome results in “leaky gut syndrome”, which could allow bacteria, toxic digestive metabolites, bacterial toxins, and small molecules to “leak” into the bloodstream and act through the gut-brain axis (GBA), facilitating the development of obesity and several other ailments . AntibioAs different classes of antibiotics produce different modifications to the microbiome, it might be suspected that obesity-related dysbiosis is attached to certain classes of antibiotics, particularly in childhood obesity ,20.Based on the above associations, we suspected that different classes of antibiotics might promote or inhibit the development of obesity-related dysbiosis, and the dominant consumption of “promoting” or “inhibiting” antibiotics in European countries might influence the prevalence of obesity in children and adults of the given countries. We aimed to identify antibiotic classes that might promote or inhibit obesity-related dysbiosis.To evaluate the relevance of our hypothesis, we have compared the antibiotic consumption data of major antibiotic classes to adult and childhood obesity prevalence Standard Protocol Approvals, Registrations, and Patient Consent: Publicly available, already published statistical data on antibiotic consumption (ECDC) and the prevalence of childhood/adult obesity prevalence (Obesity Atlas) had been compared and statistically analyzed, which did not require any ethical approval. The study did not contain any patient data, and no patient consent was requested.http://s3-eu-west-1.amazonaws.com/wof-files/11996_Childhood_Obesity_Atlas_Report_ART_V2.pdf, accessed on 22 March 2022), and the data for adults were extracted from http://gamapserver.who.int/gho/interactive_charts/ncd/risk_factors/obesity/atlas.html (accessed on 22 March 2022).Antibiotic consumption data were extracted and calculated as the average yearly consumption from the ECDC yearly reports of antibiotic consumption in the community for 30 European countries, which regularly reported consumption data to the ECDC, from 2008 to 2018. The total average yearly consumption of antibiotics for systemic use (J01) was expressed in terms of Defined Daily Dose/1000 Inhabitants/ Day (DID), and the relative share of the consumption of major antibiotic classes has been calculated in terms of the percentage of the total consumption at Anatomical Therapeutic Chemical classification (ATC) level three and level four within the group of J01C (penicillin). The major classes included were tetracycline (J01A), penicillin (J01C), broad-spectrum, beta-lactamase-sensitive penicillin (J01CA), broad-spectrum, beta-lactamase-resistant combination penicillin (J01CR), narrow-spectrum, beta-lactamase-sensitive penicillin (J01CE), narrow-spectrum, beta-lactamase-resistant penicillin (J01CF), cephalosporin (J01D), macrolides (J01F), and quinolones (J01M), covering the majority (over 90%) of the antibiotics used in the community. Obesity prevalence data for children 5–9 years of age were extracted from the Atlas of Childhood Obesity 2019 October , and we considered a non-significant (moderate) correlation when the p values remained between 0.05 and 0.09 (0.05 ≤ 0.09).Statistical analysis: Pearson’s chi-square test was applied to calculate correlations. A significant correlation (positive/negative) was estimated when p values were equivalent to or less than 0.05 (≤0.05).Countries were divided into three groups based on their rankings of childhood and adult obesity and to perform variance analysis. A one-way ANOVA test was performed to estimate a statistically significant difference between the obesity groups. A significant difference was estimated when p values were equal to or less than 0.05 (≤0.05).Childhood obesity data were divided into two groups based on means, creating below-mean and above-mean categories. The same methodology was followed for adult obesity. Logistic regression analysis was performed to determine the odds ratio  for each antibiotic class for both childhood and adult obesity. A significant result was estimated when Spreadsheets containing the antibiotic consumption databases and the obesity prevalence r: 0.517, p: 0.003), broad-spectrum, beta-lactamase-resistant combination penicillin , cephalosporin , and quinolone . A negative (“inhibitor”) correlation has been observed between childhood obesity and tetracycline , broad-spectrum, beta-lactamase-sensitive penicillin , and narrow-spectrum, beta-lactamase-sensitive penicillin . A moderate, negative association was observed with the narrow-spectrum, beta-lactamase-resistant penicillin   correlation between childhood obesity and the total consumption of antibiotics for systemic use in the community  .The rank order of countries  shows thp: 0.009) and quinolone . We have found a statistically significant difference between macrolides  and adult obesity as well.The groups formed by these ranks have been studied with one-way ANOVA to demonstrate the differences in antibiotic consumption between them. In childhood obesity, we found a statistically significant difference in beta-lactamase-resistant combination penicillin  and quinolone . On the other hand, there is a statistically significant decreased risk in the case of tetracycline . No similar association was found between antibiotic consumption and adult obesity.The logistic regression analysis showed a statistically significant elevated risk with the consumption of beta-lactamase-resistant combination penicillin  play a definite role, but the exact pathology is not appropriately identified. The most frequent explanation for obesity is the consumption of “junk food” and the lack of physical activity, but this does not explain the more-than-two-fold difference in the prevalence of childhood obesity rates in European countries . The difference for adults is lower .Experiments demonstrated the role of the microbiome in the development of obesity and diabetes. It has been observed that lipopolysaccharide (LPS), which is produced by certain bacteria, is also capable of inducing “metabolic endotoxemia”, and this effect does not occur in the absence of CD14 cells in genetically modified mice ,22.The gut microbiome plays an important role in energy homeostasis, regulating energy harvesting, fat deposition, as well as feeding behavior, and appetite. Feeding, appetite, and energy expenditure are under the control of the central nervous system (CNS), which receives various peripheral signals of energy status and its availability, such as gut hormones and adipokines  ,24.Apart from the orexigenic hormones (appetite stimulants), such as ghrelin, neuropeptides, etc., the hypothalamus regulates the homeostatic feeding behavior, while other neural brain regions, such as the insular cortex, orbitofrontal cortex, nucleus accumbens, amygdala, and dopaminergic ventral tegmental area, present groups of neurons implicated in the reward-related, non-homeostatic control of feeding .Food intake is mainly regulated by the energy needed from the brain based on its adenosine triphosphate (ATP) disposability and dysfunctions in any part of this metabolic pathway, such as congenital leptin deficiency, which can result in a persistent state of positive energy balance and obesity development ,27.The eubiotics/dysbiosis condition of the gut microbiota strongly influences our health and disease status. As the gut microbiota is a crucial factor in human physiology, many of these effects are mediated by metabolites that are either produced by the microbes or derived from the transformation of environmental or host molecules .Our study identified major classes of antibiotics , which might contribute to the development of obesity-promoting dysbiosis in childhood obesity and, probably, macrolides (J01F) in adults through different molecular mediators and GBA, which might “promote” the development of obesity. In the case of the total consumption of antibiotics for systemic use, it might be considered that the majority of the classes of antibiotics consumed belong to the “promoter” group ; this is why, in this category (J01), a positive correlation was observed. At the same time, we have found antibiotic classes that might inhibit the development of obesity-promoting dysbiosis . Antibiotic classes showing negative associations with obesity figures  might inAs J01CR  and J01CA  classes of antibiotics promote and inhibit the development of childhood obesity, respectively, the role of added beta-lactamase inhibitors  changes the direction of the effect of the antibiotics.In adult obesity, only ANOVA analysis indicated a positive correlation between the consumption of macrolide antibiotics and adult obesity data (BMI ≥ 30).Animal studies and meta-analyses of human studies on the association between antibiotics and the subsequent development of obesity are suggestive of a link between exposure to antibiotics—particularly, early exposure in life—and the development of subsequent obesity as a result of alterations in the diversity of gut microbiota. The evidence is strong in animal models, whereas evidence in humans is inconclusive, requiring well-designed, long-term longitudinal studies to examine this association . ExperimIn one of our previous publications , we obseThe restoration of the microbiome in obesity through fecal microbiome transfer (FMT) could be considered a therapeutic option , but theCurrent evidence suggests that disruptions to the composition and maturation of the gut microbiota caused by antibiotic use in early life are a key mechanism linking the association between antibiotics and obesity .The limitations of our study are that the results could not be supported at the individual level; only statistical correlations and concordance could be established. However, our findings are in accord with the reports featured in several publications.Our comparative analyses identified the classes of antibiotics that might promote or inhibit the development of obesity through the alteration of the microbiome. Mediator molecules arising from the dysbiotic microbiome, acting through the gut-brain axis, could influence the prevalence of obesity. This observation might explain the origin of the obesity pandemic.


During the COVID-19 pandemic, an Internet-Mindfulness-Based Stress Reduction (iMBSR) program was delivered and may be better than an in-person approach. Our study evaluated the effects of iMBSR intervention on mental health, self-efficacy, and body image in women with breast cancer in Taiwan.n = 41) program or a waitlist control group (n = 26), without heterogeneity between group characteristics. Patients from both groups were measured at baseline and postintervention using three scales: Depression, Anxiety, and Stress Scale (DASS-21), General self-efficacy scale, and Body Image Scale. Descriptive dataset analysis, paired t-test, and Student’s t-test were used to evaluate the data.Sixty-seven women with breast cancer were allocated to a 6-week iMBSR (p = 0.041) with medium effect sizes. Significant benefits were found for body image  and self-efficacy , with large effect sizes (Cohen’s d = 0.73).Although iMBSR did not significantly improve depression and stress between groups, iMBSR could improve anxiety (Δmean: −2.0 vs. −0.4, Our preliminary study supports iMBSR as a program that can improve mental health, body image, and self-efficacy in women with breast cancer. During the COVID-19 pandemic, medical professionals can use Internet-based clinical health education. A 2019 cancer registry report indicated that breast cancer had the second highest mortality rate in Taiwan and the highest incidence among cancers in women and increased by ~4.49% per year .Adverse effects of breast cancer treatment, such as cognitive impairments , alopeciBased on Bandura’s theory, self-efficacy refers to the ability of an individual to control their motivation, behavior, and how they face obstacles. The theory essentially attributes an individual’s belief in efficacy to their chance of achieving success in a particular situation . TherefoStudies have highlighted that mindfulness-based interventions have a wide range of benefits, including decreased perceived stress, anxiety, depression, and fatigue , improveWith the outbreak of COVID-19 in December 2019  and its Patients with breast cancer are at higher risk of psychological stress, and the additional stress of the COVID-19 pandemic can increase their vulnerability . InterneA study in Denmark involving women with breast cancer and older men with prostate cancer who underwent an Internet-delivered MBI (iMBI) program found that iMBIs could improve anxiety and depressive symptoms immediately after the intervention . AnotherConsiderable research explores the effects of iMBSR on physical and psychological problems in patients without cancer, and using iMBCT to resolve emotional distress in patients with cancer. However, there has been little research on iMBSR’s effects on self-efficacy and body image in women with breast cancer. Therefore, our study aimed to explore the effect and feasibility of iMBSR on mental health, self-efficacy, and body image in women with breast cancer during COVID-19.Both the iMBSR and waitlist control group used purposive and snowball sampling. Potential participants were recruited from visiting breast surgery clinics, cancer wards, and breast cancer affiliate websites. Inclusion criteria were as follows: had been diagnosed with stage 0–IV breast cancer within the past 5 years; aged ≥20 years old; could communicate in Mandarin; received at least recent adjuvant therapy ; and Eastern Cooperative Oncology Group performance score ≤ 1 . ExclusiOur study has several required online software to facilitate online teaching and recruiting participants . The recAll participants submitted written informed consent during the orientation. Variable measurement was performed at two time points, including pretest and posttest. (1) Pretest: This was completed before the program’s second session. Baseline scores were based on the participants; responses to items across the scale. (2) Posttest: After the sixth week of training, participants completed a posttest within a week. Participants received a TWD 100 electronic coupon after completing the pre and postintervention questionnaires.The frequency and duration of our mindfulness training were six 2-h weekly group coaching sessions led by a professionally trained psychologist using online learning with Microsoft Teams. We used Jamboard, a digital interactive whiteboard developed by Google, to measure the number of patients participating online before class. Our training aimed to improve these metrics: mindful eating, body scan, breath awareness practice, mindful walking, and sharing of group experiences. The informal training in our study was conducted through home practice using mindfulness-guided cloud-based applications such as YouTube . SpecifiInclusion and exclusion criteria for WCG (delay in receiving the intervention) were the same as for the iMBSR group. WCG occurred in parallel with iMBSR. The WCG had manuals for managing symptoms and oral instructions on health and hygiene education. All participants were instructed not to participate in any other stress reduction program or mind–body therapy during the study. We offered an additional iMBSR skills study program to WCG participants following our study’s conclusion.The present study involved research with human participants who were required to sign an informed consent form before participating in the study. All procedures in our research comply with the ethical standards of the Institutional Research Council.n = 356) and compared with n = 720). Multi-group confirmatory factor analysis found that the Chinese version of the DASS-21 effectively discriminates between depression, anxiety, and stress, but less so than the English version.The Depression, Anxiety, and Stress Scale (DASS-21) is a 21-item tool that evaluates the participant’s depression, anxiety, and stress over the past week, on a Likert-style scale ranging from 0 (never) to 3 , with sensitivity to change  and good discriminant validity , developed by Jerusalem and Schwarzer, is a generic tool that is widely used to measure self-efficacy in clinical and nonclinical populations . Zhang eThe questionnaire was divided into two parts. First: basic personal information, including age, marital status, and economic status. Second: disease status, including time since diagnosis (years), cancer staging, and cancer treatment. The questionnaire was collected using an online Google form.To maintain the fidelity of our online mindfulness program, the 6-week intervention was conducted by a qualified clinical psychologist with specialized training in psychology at the Medical Center, Departmental Cancer Center. The instructor assessed the transfer of iMBSR skills by asking questions and discussing the material with the participants, and the patients recorded their practice duration weekly. The first author (YCC) confirmed that the practice had been completed during the intervention. We also prepared a standardized process. First, we set up a LINE messenger group with a mobile app before iMBSR to remind participants of the class schedule, provide class information, and solve the participants ‘problems during the course . Second, we regularly provided support and addressed participants’ needs wherever possible. Third, we used the patient groups to care for each other and share experiences . Finally, if the participants completed the program, we provided an e-coupon as a reward.The sample size of our study was calculated based on the results of our previous study . We assut-tests and Student’s t-tests for normally distributed continuous variables, respectively. The statistical significance level was set to a two-sided p < 0.05 in all statistical analyses.Data analysis was performed using SPSS for Windows 22.0 . The collected questionnaires were coded, and the accuracy of the data was verified repeatedly after input. Descriptive statistics were described for the baseline demographic and clinical characteristics of the patients with breast cancer. The Kolmogorov–Smirnov test (KS test) tested raw data for adherence to the assumptions of normality and equal variance and the detection of outliers before inferential statistical procedures. Self-report data between baseline and 6 weeks were compared for within-group and between-group data analyses using paired t-test results.To explore the magnitudes of difference between groups, Cohen suggested that effect sizes be categorized into small, medium, and large treatment effects and represented by 0.2, 0.5, and 0.8, respectively . A websin = 15) were lack of IT skills, physical discomfort, scheduling conflict, and others. We had established baseline data for 72 participants before the first session, with six dropouts in the WCG and three who changed to the Internet format. The postintervention assessment was completed by 41 and 26 participants in the MBSR and WCGs, respectively. The demographic and clinical characteristics of the patients are listed in p = 0.043).Our study had 88 eligible participants (see p = 0.041). However, depression , stress , and overall DASS-21 scores  were not significantly decreased. Six consecutive weeks of iMBSR sessions also improved body image  and self-efficacy  in women with breast cancer.We used DASS-21 to measure participants’ current psychological problems. The DASS-21 results indicated significantly improved anxiety  and significant decreases in the subscale of depression , anxiety , and stress . The intervention significantly improved the participants’ negative body image  and increased self-efficacy  found improvements in anxiety and depression in women with breast cancer following Internet-based Mindfulness-Based Cognitive Therapy  at risk of coronavirus infection , therebyOur study has several strengths. Our investigation was during the COVID-19 pandemic, highlighting the importance of an Internet-based psychoeducational intervention. The e-delivery prevented exhaustion from long commutes and reduced infection rates, which was critical for patients with cancer and potentially compromised immune systems. Additionally, we initially expected to enroll 48 participants, but we completed the final program with 67, exceeding our expected inclusion rate. However, this study had some limitations. First, we did not measure the long-term effects of iMBSR on patients with breast cancer. Second, the average age of the participant was 49.61 ± 12.03. Some older participants struggled with the Microsoft Teams software and could not join the course, hear the sound, or operate the digital whiteboard. Therefore, the staff should have spent more time contacting individuals struggling with the software.Overall, our findings indicate that we provided women with breast cancer with a convenient and cost-effective method to enhance their empowerment and confidence through mindfulness-based mental health education that taught them how to easily incorporate mindfulness techniques into their daily lives. We propose a future study investigating the acceptability and cost-effectiveness of Internet-based mindfulness interventions for people with different sociodemographic conditions.The original contributions presented in the study are included in the article/supplementary material; further inquiries can be directed to the corresponding authors.The studies involving human participants were reviewed and approved by Research Ethics Committee, China Medical University & Hospital, Taichung, Taiwan. The patients/participants provided their written informed consent to participate in present study.Y-CC: study conception and design. Y-CC, C-FC, C-KW, C-TW, L-CL, and Y-CW: data collection and critical revision of the article. Y-CC and C-KW: data analysis and interpretation and drafting of the article. All authors contributed to the article and approved the submitted version.This research was partially supported by a grant from China Medical University, Taiwan (CMU110-N-11 and CMU111-MF-115) and Ministry of Science and Technology, Taiwan (MOST 110-2314-B-039-059 and MOST 111-2314-B-039-016), received by Y-CC.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


In March 2020, Scottish government identified people clinically extremely vulnerable to COVID due to pre-existing health conditions. These people were advised to strictly self-isolate (shield) at home. We examined who was identified as clinically extremely vulnerable, how their healthcare changed during isolation, and whether this process exacerbated healthcare inequalities.We linked all individuals on the shielding register in NHS Grampian to their in-patient and out-patient healthcare records from 2015 through 2020. We analysed the method of patients’ identification as clinically extremely vulnerable . We measured out-patient, in-patient, and emergency healthcare attendances, and compared use rates between two 3-month periods before and during the first strict isolation period. We evaluated changes in care use between those shielding and the general non-shielding population, and differences between shielding sub-populations .The shielding register included 16,092 people (3% of the population). 42% of people on the register were not identified by national healthcare record screening, including the majority of cancer and immunocompromised patients. People added to the register by their care-providers were more likely to be young and less economically-deprived.Shielders’ healthcare use decreased during isolation . However, people shielding had better maintained care than the non-shielding population (e.g. RR 2.9 for scheduled in-patient care). There were inequalities in whose scheduled care was maintained while shielding: younger people and those with cancer had significantly higher visit rates. However, there were no differences in care-preservation between men and women or between socioeconomic deprivation levels.The reliance on emergency care while shielding indicates that, overall, continuity of care for existing conditions was not optimal. However, there was notable success in maintaining care for cancer. We suggest that integrating demographic and primary care data would improve identification of the clinically vulnerable and help equitably prioritise care.


NBS-LRR genes are the largest gene family in plants conferring resistance to pathogens. At present, studies on the evolution of NBS-LRR genes in angiosperms mainly focused on monocots and eudicots, while studies on NBS-LRR genes in the basal angiosperms are limited. Euryale ferox represents an early-diverging angiosperm order, Nymphaeales, and confronts various pathogens during its lifetime, which can cause serious economic losses in terms of yield and quality. In this study, we performed a genome-wide identification and analysis of NBS-LRR genes in E. ferox. All 131 identified NBS-LRR genes could be divided into three subclasses according to different domain combinations, including 18 RNLs, 40 CNLs, and 73 TNLs. The E. ferox NBS-LRR genes are unevenly distributed on 29 chromosomes; 87 genes are clustered at 18 multigene loci, and 44 genes are singletons. Gene duplication analysis revealed that segmental duplications acted as a major mechanism for NBS-LRR gene expansions but not for RNL genes, because 18 RNL genes were scattered over 11 chromosomes without synteny loci, indicating that the expansion of RNL genes could have been caused by ectopic duplications. Ancestral gene reconciliation based on phylogenetic analysis revealed that there were at least 122 ancestral NBS-LRR lineages in the common ancestor of the three Nymphaeaceae species, suggesting that NBS-LRR genes expanded slightly during speciation in E. ferox. Transcriptome analysis showed that the majority of NBS-LRR genes were at a low level of expression without pathogen stimulation. Overall, this study characterized the profile of NBS-LRR genes in E. ferox and should serve as a valuable resource for disease resistance breeding in E. ferox. Euryale ferox (prickly waterlily) is an annual aquatic plant in tropical and subtropical regions of Southeast and East Asia and is the only species in the Euryale genus in the Nymphaeaceae family. It is cultivated as a nutritious food due to its high starch content; additionally it is also a common Chinese traditional medicine which can treat depression and diabetes mellitus  have been cloned from E. ferox.mellitus . HoweverR genes in plants, which play a core role of the immune system, detect pathogens and trigger downstream resistance response, frequently accompanied by hypersensitive reactions (HR) (NBS-LRR) genes are the largest R gene family in plants. To date, more than 60% of the functional resistance genes cloned from angiosperms belong to the NBS-LRR gene family, which can provide resistance to bacteria, fungi, viruses, nematodes, and other pathogens  with different domain combinations along plant evolution (NBS-LRR genes can be divided into three subclasses: TIR-NBS-LRR (TNL), CC-NBS-LRR (CNL), and RPW8-NBS-LRR (RNL)  activation, so it was called ‘helper NBS-LRR’ (hNLR)  . The nucathogens . NBS-LRRvolution . AccordiRR (RNL) . The majetectors . NBS-LRRT domain . After rferation . The low’ (hNLR) . Furtherignaling . Recent ll death .NBS-LRR in Arabidopsis thaliana and rice early in the 21st century, NBS-LRR genes have been identified and analyzed in dozens of plant genomes  database (https://genomevolution.org/CoGe/GenomeInfo.pl?gid=56574). The RNA-seq data of E. ferox was downloaded from the National Center for Biotechnology Information (NCBI) database (https://www.ncbi.nlm.nih.gov/sra/?term=SRR9650346). The NBS-LRR genes in Nymphaeaceae colorata and N. thermarum were downloaded from the ANNA database (http://compbio.nju.edu.cn/app/ANNA/) (The genome sequence and annotation files of p/ANNA/) .NBS-LRR genes were identified in the E. ferox genome as described previously (E. ferox with the amino acid sequence of the NB-ARC domain (Pfam: PF00931) as a query, with a threshold expectation value of 1.0. Meanwhile, BLASTp search was performed towards the protein sequences of E. ferox using the sequence of the HMM profile of the NB-ARC domain . Then, all hits obtained using the two methods were merged, and the redundant hits were removed. In order to confirm that blast hits contained the NBS domain, a round of HMMscan was conducted for all the obtained proteins using a more strict threshold expectation value . All non-redundant candidate sequences were submitted to an online tool—the Conserved Domains Database at NCBI  to further verify the CC, TIR, RPW8, LRR, and other integrated domains.eviously . BrieflyNBS-LRR genes to specific positions on chromosomes of the E. ferox genome, the annotation file recording the positions of all genes was used, and the genomic locations of all the NBS-LRR genes were extracted from the file. The flanking regions of each NBS-LRR gene  were searched for the presence/absence of other NBS-LRR genes. If another NBS-LRR gene was detected within the flanking regions, the two NBS-LRR genes were considered as located in the same NBS-LRR gene cluster    were ide studies , which mu et al. . NotablyNBS-LRR genes possess the entire structure, and some NBS-LRR genes may lose part of their domains or fuse additional domains during duplication  structure protein. The structures of TNL and CNL genes are both more diverse than those of RNL genes. We explored 13 different kinds of structures in 73 TNL genes and seven kinds of structures in the 40 CNL genes. Notably, these structural variations were caused not only by the loss of the typical domains of NBS-LRR genes but also by the fusing of additional domains. For example, we found four different integrated domains (IDs) in nine TNL genes. Among them, seven IDs were fused to the C-terminal domain of NBS-LRR proteins, whereas two were fused to the N-terminal domain. Moreover, transcription analysis indicated that the majority of NBS-LRR genes of E. ferox were expressed at a low level  and 18 multigene clusters , which were distributed mainly on chromosomes 11, 15, and 18 . The topology was consistent with that of other investigated angiosperms, supporting the notion that the three subclasses diverged in early ages  were recovered (NBS-LRR genes from E. ferox and two water lilies were detected in the phylogenetic tree, which mirrored the species-specific expansion, especially the drastic expansion of CNLs in N. colorata in the C44 lineage, with 102 CNLs. Of 122 ancestral lineages, E. ferox inherited 70 lineages, whereas there were only 131 NBS-LRR genes in the E. ferox genome, suggesting that a fairly low level of NBS-LRR gene expansion occurred in the E. ferox genome during evolution.Based on the reconstructed ecovered . ClusterNBS-LRR genes consist of hundreds of members and are the largest family among R genes. In two-tier immune systems, the NBS-LRR gene family plays important roles in effector-triggered immunity. Revealing the profile of NBS-LRR genes in a species is very important, and by exploring the evolutionary patterns of NBS-LRR genes in the species and its close relatives, we can put these conclusions into practice (NBS-LRR gene family (NBS-LRR genes: TNLs, CNLs, and RNLs. Studies have shown that proteins encoded by CNLs and TNLs function as receptors for pathogen recognition, and these two subclasses have expanded to various degrees during the long-term ‘arms race’ with pathogens (practice . In the practice . It is ne family . Angiospathogens .E. ferox genome was found to harbor only 131 NBS-LRR genes, consistent with a recent report. NBS-LRR genes occurred during the evolution of adaptation to an aquatic lifestyle, which may be the reason for the fewer NBS-LRR genes in E. ferox compared to other angiosperms. The structures of NBS-LRR genes are relatively diverse. Among four different IDs, MARCH1, also known as membrane-associated RING finger protein 1, is a membrane-anchored E3 ubiquitin ligase that mainly expressed in cells of the immune system (In this study, the e system . MARCH1 NBS-LRR genes can expand via different duplication methods. Many studies have shown that tandem duplication is dominant in NBS-LRR gene expansion in most angiosperms (NBS-LRR genes in the E. ferox genome and found that there were 28 NBS-LRR genes generated by tandem duplication in the E. ferox genome, accounting for only 21% in total. This proportion is much lower than the 70–80% in the legume family (E. ferox experienced two rounds of WGDs (E. ferox NBS-LRR genes are derived from segmental duplications, which is different from the situation in most other angiosperms, and the fewer number of tandemly duplicated genes may also be one of the reasons for the relatively fewer NBS-LRR genes in E. ferox. Additionally, the phylogenetic analysis of NBS-LRR genes in E. ferox and two other Nymphaeaceae species revealed that there were at least 122 ancestral NBS-LRR genes in the common ancestor of the three species, whereas only 131 NBS-LRR genes were found in the E. ferox genome, which further indicated a low level of NBS-LRR gene expansion in the E. ferox genome during its long-term evolution.iosperms . We anale family . Further of WGDs . Thus, wNBS-LRR genes from genomic data is not only important for revealing the evolution of this important disease resistance gene family but also serves as an important basis for discovering and utilizing functional disease resistance genes in crops. Based on the analysis of NBS-LRR genes in the rice genome, NBS-LRR genes in resistant rice varieties, and verified dozens of NBS-LRR genes that play important roles in blast resistance in rice. Solanum americanum by target region sequencing and third-generation sequencing based on the analysis of NBS-LRR genes in the potato genome. These studies indicate that the genome-wide identification and analysis of NBS-LRR genes promotes the cloning of functional resistance genes in the studied species and related species. E. ferox is an important functional food and medicinal plant that has been widely cultivated in China since time immemorial. However, due to its characteristics of underwater flowering and pollination, research on germplasm innovation and disease resistance breeding has lagged behind research in other species. With the breakthrough of artificial hybrid technology for E. ferox in the 1980s and the development of tissue culture technology in recent years, it is possible to breed superior varieties of E. ferox by molecular breeding. E. ferox is vulnerable to a large number of diseases, so it is particularly important to develop molecular breeding in E. ferox.The comprehensive identification of NBS-LRR genes in the genome of E. ferox. Although the quantity is significantly lower than that of most species in Solanaceae, Leguminosae, and Poaceae, it is roughly equivalent to that of Brassicaceae crops, and higher than those of Cucurbitaceae crops (NBS-LRR genes, analysis of sequence features, and revelation of chromosomal distribution pattern will provide important references for the molecular breeding of E. ferox. In addition, Nymphaeaceae contains many kinds of important flowering plants. The comparative analysis of NBS-LRR genes in these plants will also facilitate the molecular breeding of different species in the same genus.In summary, the present study identified a complete set of 131 ae crops . The ide


The scientific literature regarding Mott cell differentiation in canine lymphoma is scarce. Mott cells are defective in immunoglobulin secretion and are derived from plasma cells, and lymphoma is a severe condition characterized by the proliferation of neoplastic lymphoid cells. Lymphoma can be divided into B- or T-cell according to their origin. Whether the origin of lymphoma is B- or T-cell can be confirmed by PCR for antigen receptor rearrangement or flow cytometry assay. However, the phenomenon in which B- and T-cells are simultaneously identified in PCR for antigen receptor rearrangement and flow cytometry is called cross-lineage rearrangement and lineage infidelity, respectively, and is known to be occasionally found in canine lymphoma. These phenomena have not been reported in canine lymphoma with Mott cell differentiation. This study is the first report of Mott cell differentiation in canine B-cell lymphoma with cross-lineage rearrangement and lineage infidelity. This study describes the clinical features, diagnosis, and treatment of this unknown type of cancer in a 4-year-old female mongrel dog.Lymphoma is a severe condition characterized by the proliferation of neoplastic lymphoid cells. A 4-year-old female mongrel dog presented with solitary lymph node enlargement. Significant right prescapular lymphadenopathy and abdominal enlargement were observed during physical examination. A complete blood count revealed lymphocytosis, and a peripheral blood smear revealed lymphoblastosis and Mott cells. Fine needle aspiration cytology (FNAC) of the right prescapular lymph node revealed a predominant population of lymphoblasts and Mott cells. Based on the FNAC and blood smear results, the patient was diagnosed with leukemic state multicentric B-cell lymphoma with Mott cell differentiation. Subsequent PCR for antigen receptor rearrangement and flow cytometry revealed that the patient exhibited cross-lineage rearrangement (CLRA) and lineage infidelity (LI), respectively. CHOP-based chemotherapy was initiated, however, the patient’s disease was progressive. The patient died three months after the initial presentation. Mott cell differentiation in canine B-cell lymphoma (MCL) has rarely been reported in the veterinary literature and seems to show an unusual clinical course. To the best of our knowledge, no reports of MCL with CLRA and LI exist. We report the clinical features, diagnosis, and treatment of MCL with CLRA and LI. Lymphoma  is the uncontrolled proliferation of neoplastic lymphoid cells that arises in lymphoid- or other tissues in the body. It is commonly encountered in small animal practices and comprises approximately 83% of canine hematopoietic cancers . No sex Mott cells are defective in immunoglobulin secretion and are derived from plasma cells ,10,11. T2) and abdominal enlargement. Complete blood count  revealed severe lymphocytic leukocytosis  and mild anemia   . A blood–57.0%]) .TM, Shenzhen, China) were unremarkable. Abdominal ultrasonography detected prominent intra-abdominal lymphadenopathy, ascites, and a splenic mass with a honeycomb sign  using CBC and a refractometer, and cytology revealed predominant lymphoid cells, consistent with the features of neoplastic effusion .A PARR  assay using the right prescapular lymph node (LN) revealed that the patient presented with clonal rearrangement of both IG and T-cell receptor genes, a phenomenon known as CLRA.Immunophenotyping via FC assay was performed on right prescapular LN aspirates and peripheral blood samples. In order to select the best gating strategy for the determination of the lymphoid cells in the right prescapular LN and peripheral blood, lymphoid cells were identified and gated by forward scatter (FSC) and side scatter (SSC) characteristics. After doublet exclusion of the gated lymphoid cells, the singlets were gated . Then, i2, IV), vincristine , doxorubicin , and prednisolone , was initiated. The patient was monitored weekly for clinical response to chemotherapy throughout the protocol, and abdominal ultrasonography was repeated at the end of each chemotherapy cycle.Based on the results of FNAC, PARR, FC, and the presence of Mott cells, the patient was diagnosed with stage Va  multicentric B-cell LSA with Mott cell differentiation, CLRA, and LI. CHOP-based chemotherapy using cyclophosphamide (250 mg/m9 cells/L).The initial response to chemotherapy was favorable during the first cycle of CHOP therapy. The enlarged superficial and intra-abdominal LNs decreased in size by approximately 40%, and lymphocytosis on CBC was within the reference interval  and prednisolone  was initiated after L-CHOP chemotherapy at the request of the owner. Unfortunately, the response to this treatment was also consistently poor, and superficial and intra-abdominal LNs and the spleen gradually increased in size until the final visit  was not achieved, and partial remission (PR) was maintained for the first four weeks. In the 6th week of chemotherapy , the patient’s superficial and intra-abdominal LNs and spleen were observed to be enlarged, which was considered a progressive disease (PD), followed by CHOP chemotherapy and L-asparaginase administration . The response to re-instituted chemotherapy over four weeks was consistently poor, and palliative therapy using chlorambucil  and FC analysis of peripheral blood (immunophenotyping of CD3−/CD5−/CD21+/CD34−). Mott cells are derived from terminally differentiated B cells, and neoplastic lymphoid differentiation combined with Mott cell differentiation is indicative of B-cell LSA . These rIn the present patient, CLRA and LI were found using PARR and FC, respectively. PARR is a methodology used to detect clonality in B-cell and T-cell LSA . NormallFC is a useful test for diagnosing immunophenotype LSA and has the advantage of providing a larger panel of markers than IHC or ICC . It is hAlthough rarely curable, most LSAs are initially responsive to treatment, and LSA that responds to chemotherapy has a better prognosis than LSA that does not ,8,40. InTo our knowledge, this is the first case report of MCL with CLRA and LI. The patient’s response to therapy remained consistently poor despite treatment with CHOP-based chemotherapy. Taken together, MCL might be considered a negative prognostic factor and appears to have an unusual clinical course in canine B-cell LSA. A complete characterization of this type of LSA requires further investigation with additional case studies, and further studies on the clinical implications of CLRA and LI are required.


Roadside testing of illicit drugs such as tetrahydrocannabinol (THC) requires simple, rapid, and cost-effective methods. The need for non-invasive detection tools has led to the development of selective and sensitive platforms, able to detect phyto- and synthetic cannabinoids by means of their main metabolites in breath, saliva, and urine samples. One may estimate the time passed from drug exposure and the frequency of use by corroborating the detection results with pharmacokinetic data. In this review, we report on the current detection methods of cannabinoids in biofluids. Fluorescent, electrochemical, colorimetric, and magnetoresistive biosensors will be briefly overviewed, putting emphasis on the affinity formats amenable to on-site screening, with possible applications in roadside testing and anti-doping control. Cannabis sativa is an annual flowering plant consisting of several botanical variants that produce terpenes, fatty acids, and flavonoids, alongside the major compounds, the cannabinoids. Over time, cannabis has been used for both therapeutic and recreational purposes because cannabinoids are involved in many physiological processes in animals and plants. [9-tetrahydrocannabinol (THC) and cannabidiol (CBD), the decarboxylated forms of Δ9-tetrahydrocannabinolic acid (THCa) and cannabidiolic acid (CBDa) are the most sought after by consumers, due to their psychoactive and therapeutic effects. plants. . Cannabi plants. . More th plants. . Among t-Cannabinoids that are produced within the body,  ,4;-Phytocannabinoids  such as THC and CBD ;-Synthetic cannabinoids that are hallucinogenic chemicals ,6.There are three types of cannabinoids: 1R) [1R and type-2 cannabinoid receptor (CB2R), although with lower affinities, compared to THC [It has been almost three decades since THC has been used for curative purposes. Various cannabis extracts with settled THC/CBD ratios, as well as pharmaceutical-grade herbal cannabis, have been reported in the treatment of several conditions . The pha1R) . CBD intd to THC . The regd to THC . The othd to THC ,12. Targ9-tetrahydrocannabinol (THC-OH) in the liver (by cytochrome P450), followed by the oxidation of THC-OH to 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid (carboxy-THC) [The metabolic route of THC starts with its hydroxylation to 11-hydroxy-Δoxy-THC) . Thus, Toxy-THC) . An indioxy-THC) . The mosAlthough CBD is non-psychoactive, it has been extensively used for its validated benefits in the treatment of various health issues such as epilepsy, chronic pain, insomnia, and addiction ,16. The 1 and CB2 receptors, thus causing psychoactive effects usually obtained with THC [The significant growth of cannabis consumption has heavily impacted the synthetic cannabinoids (SCs) markets. SCs belong to a class of chemicals from the new psychoactive substances (NPS) group, being marketed as natural herbal mixtures under various brand names ,20. SCs with THC . The abuwith THC . The firwith THC . Newly rwith THC . The EU with THC .Cannabinoids are listed as substances whose use is prohibited in sports competitions along with performance-enhancing substances (PES). Anabolic substances that stimulate muscle growth, and substances that enhance oxygen transport are also included in the PES list ; CannabiThe established detection method of cannabinoids in authorized laboratories is gas chromatography-tandem mass spectrometry GS-MS/MS. However, the GC-MS/MS methods require complex sample derivatization steps and long analysis times . In addiOverall, roadside testing requires (a) the use of portable, easy-to-handle, and miniaturized equipment (b) non-invasive sample collection with minimum risk of contamination, and (c) adequate analytical methods . As in the case of point-of-care (POC) applications, these systems must meet the ASSURED  criteria. Electrochemical sensing is by far a technique able to match the ASSURED conditions with respect to cannabinoid detection in body fluids. THC detection is based on its electrochemical oxidation, following a reaction path similar to phenol . There aThe testing manner is heavily dependent on the cannabinoids’ pharmacokinetics. THC appears right away in blood following the first inhalation, with the TCH level reaching a peak within 8 min. Almost 65% of cannabis is eliminated via feces as TCH-OH, while 20% is excreted in urine as carboxy-THC and its glucuronic acid conjugate . The phaOral fluid  contains more than 97% water, plus electrolytes, epithelial cells, white blood cells, enzymes  immunoglobulins, and glycoproteins (mucin) . Unlike Exhaled breath contains a multitude of volatile organic compounds (VOCs) in the vapor phase, and non-volatile compounds impregnated in the suspended solid particles . THC is Sweat contains 99% water, plus salts, fat, ammonia, urea, and sugars; overall, there are fewer impurities than in other biofluids. Sweat is less prone to degradation . Sweat iCollecting blood samples is an invasive procedure that requires skilled personnel. Electrochemical sensors can detect simultaneously THC and its metabolites in a single blood sample. The ratios of cannabinoid concentrations provide information about the timeline of cannabis ingestion . The THCUrine is the most tested matrix for cannabinoid detection, being reliable, inexpensive, and easy to collect . The facIn this section, we discuss papers reporting on various methods for rapid detection amenable to on-site testing. The experimental techniques to be described here are electrochemical, fluorescence, and colorimetric. Emphasis will be put on novel strategies for surface modification, and assay principle, as both factors affect sensor efficiency. In the past decade, a new class of sensors  gain ground in the bimolecular detection in protein assays using magnetic tags .Modified surfaces can also act as filters that allow only the target analyte to participate in the electrode reaction. In the case of cannabinoids, as in the case of other drugs, the investigation of the mechanism of action within the organism is the basis of biosensor development. A drug’s mechanism of action is based upon its interaction with bio-targets such as antibodies, enzymes, ion channels, receptors, and transporters. Assays that measure the interaction between drugs and their bio-targets are carried out in vitro and may use whole cells or cellular lysates containing isolated bio-targets . Most coElectrochemical techniques are rapid and cost-efficient, widely applied and amenable to the detection of illicit drugs, such as cocaine, marijuana, and ecstasy . The eleTo overcome the drawback associated with THC’s low solubility in aqueous media or with inherent interferences caused by alcohol solvents, biosensors based on affinity recognition of THC and metabolites by immobilized biomolecules, have been recently spotted. The most relevant papers will be further discussed.In SWV, the staircase potential ramp is modified with square-shaped potential pulses . At eachKohansal et al. have developed an immunosensor for 2-AG in human plasma and rat serum exploiting the conductivity changes at the modified electrode in the presence of the ferricyanide/ferrocyanide system . The intAccording to the relation between the charge transfer and the measured parameters, EIS-based sensors can be categorized into (A) faradaic and (B) non-faradaic (capacitive) :(A) Faradaic (bio)sensors use electrodes with conductive surfaces; the charge transfer resistance (Rct) is measured in the presence of redox-active species in solution, such as hexacyanoferrate (II)/(III) anions, or hexaammineruthenium (II)/(III) cations . The dep(B) Non-Faradaic (bio)sensors are devices containing electrodes with surfaces covered by layer(s) with insulating properties. Here, the double-layer capacitance (Cdl) is the main parameter that characterizes the processes at the electrode interface . The adsN-4-hydroxypentyl metabolite) [N-4-hydroxypentyl metabolite) were determined as 10–500 ng/mL and 5.892 ng/mL, respectively. The selectivity of the biosensor was evaluated with different interfering molecules . Finally, the immunosensor successfully detected JWH-018 (N-4-hydroxypentyl metabolite) in spiked synthetic urine samples. The results showed that the developed platform can be applied to detect other JWH series with high sensitivity and accuracy.Durmus et al. proposed an EIS immunosensor for the detection of JWH-018 (abolite) . First, The affinity-based immunosensor developed by Stevenson et al. was able to detect THC in saliva . Non-far2O2, the immobilized HRP and thionine were involved in a sequence of consecutive redox reactions that led to the increase in the faradaic current. The binding of the target THC to the immobilized antibody prevented the electron transfer due to steric hindrance. Chronoamperometry was used to determine the THC in phosphate buffer saline (PBS). The results showed that the response current had a good linear correlation with the THC concentration range from 0.01 to 103 ng/mL. The LOD for THC was decreased to 3.3 pg/mL. Chronoamperometry (CA) is a potential step method: it measures the current variation vs. time as a response to a variation  in potential. Since the measurements are performed in a transient state, it allows a dramatic reduction of response time compared to amperometry in stirred solution . A sandwMagnetoresistance (MR) represents the change of resistance upon applying an external magnetic field. The capability of converting magnetic signals to electrical signals has led to the successful development of sensing devices based on the MR effect . The GMRA sensing platform that employs GMR immunosensors integrated with a portable reader system and smartphone to detect THC in saliva using competitive immunoassays was reported by Lee et al. . The comFluorescence-based detection has grown in popularity, being a swift and on-point detection technique. The popularity of this technique is probably due to its simplicity, high sensitivity, and capability of detecting analytes in small volumes. Moreover, low-cost and simple devices for on-site detection can be designed using a light-emitting-diode (LED) for excitation and photodetector . FluoresPlouffe and Murthy  reported1 and CB2 receptors was exploited to develop activity-based bioassays. A bioassay in which the cannabinoid receptor activation by cannabinoids caused the recruitment of truncated β-arrestin 2 to the cannabinoid receptors, resulting in functional complementation of a split luciferase, allowed readout via bioluminescence [LFIA results are often obtained by visualizing the color changes in specific areas, thus providing qualitative information about the presence/absence of the target analyte . Colorimnescence . This menescence , and thiAllosteric modulators are in general small molecules that can bind to a distant site  from the binding (orthosteric) site, causing a functional change of the binding site . Some alThus, carboxy-THC and CBD were detected via their interaction with the growth hormone secretagogue receptor (GHS-R1a), in competition with the labeled ghrelin GHR) (GHS-R1a’s endogenous ligand)  Figure . It was The analysis was performed on spiked urine samples and was able to detect cannabinoids even in the presence of GHR mimetics. The proposed method exploited the linear parts of the competitive binding curves to detect CBD and carboxy-THC. A linear range of 5–30 ng/mL was obtained for both CBD and carboxy-THC. A summary of biosensors for cannabinoid detection based on molecular recognition along with their performance features and drawbacks is presented in THC and its metabolite, carboxy-THC, are important targets for the assessment of cannabis use, with regards to both drugged driving (THC levels), and drug use over time (carboxy-THC levels) . Synthet


Recombinant human granulocyte colony-stimulating factor (rhG-CSF) reduces neutropenia events and is widely used in cancer patients receiving chemotherapy. However, the effects of rhG-CSF on distant organ metastasis (DOM) in non-small-cell lung cancer (NSCLC) patients following postoperative chemotherapy are not clear.A retrospective cohort study was performed on NSCLC patients who underwent complete surgical resection and postoperative systemic chemotherapy at The First Affiliated Hospital of Nanchang University between 1 January 2012 and 31 December 2017. The effect of rhG-CSF on DOM was assessed with other confounding factors using Cox regression analyses.n = 246 in the rhG-CSF group, n = 61 in the No rhG-CSF group). The incidence of DOM in postoperative NSCLC patients with rhG-CSF treatment was observably higher than in patients without rhG-CSF treatment . Univariate regression analysis revealed that rhG-CSF and pathological stage were independent risk factors for metastasis-free survival (MFS) (p < 0.05). RhG-CSF users had a higher risk of DOM  than nonusers of rhG-CSF. The association between rhG-CSF and the risk of DOM was significant only in patients presenting with myelosuppression  and not in patients without myelosuppression . The risk increased with higher dose density of rhG-CSF compared to rhG-CSF versus no users (p for trend< 0.001).We identified 307 NSCLC patients who received postoperative systemic chemotherapy (These analyses indicate that rhG-CSF use is related to DOM following postoperative chemotherapy in NSCLC. Non-small-cell lung cancer (NSCLC) is one of the most common cancers and the leading cause of cancer death worldwide . SurgeryRecombinant human granulocyte colony-stimulating factor (rhG-CSF) is commonly used for cancer patients following myelosuppressive cytotoxic chemotherapy, and it decreases the risk of potentially fatal infections and hospitalisation associated with febrile neutropenia (FN) and ensures that the dose intensity of chemotherapy improves the overall survival (OS) . HoweverHowever, both of these effects were eliminated in the presence of G-CSF inhibitors. However, clinical data rarely indicate that rhG-CSF use in cancer patients is related to a high DOM risk. The impact of rhG-CSF treatment in postoperative chemotherapy patients with myelosuppression on cancer metastasis, especially in NSCLC, is not clear. Therefore, we examined the pro-metastatic character of rhG-CSF in a retrospective cohort of NSCLC patients who received chemotherapy after surgery. These data provide experimental evidence to reduce the occurrence of cancer metastasis by pursuing G-CSF.The Ethics Committee of the First Affiliated Hospital of Nanchang University, China approved this retrospective cohort study, which abided by the rules of the Declaration of Helsinki. We reviewed the digital medical records of 307 consecutive NSCLC patients for pathological stage IB-IIIB (AJCC system 8th edition) who underwent complete surgical resection via lobectomy or greater and systematic lymph node dissection (lymph node dissection was performed depending on tumour location) and received postoperative system chemotherapy at The First Affiliated Hospital of Nanchang University between January 1, 2012 and December 31, 2017. Complete resection was defined as the macroscopic and microscopic absence of residual cancer (R0). Patients were excluded when they had been diagnosed with cancer other than NSCLC before surgery or when they underwent limited resection (segmentectomy or wedge resection), incomplete resection, neoadjuvant chemotherapy/radiotherapy, metastasis occurring before surgery or before receiving the first postoperative chemotherapy, or a follow-up time of fewer than 30 days after surgery. All patient records were anonymised before analyses. Follow-up duration was counted from the day of surgery to the day of DOM diagnosis, last follow-up, or loss to follow-up. The last follow-up date was December 31, 2020.One research group blinded to the outcome status collected clinical records data of the potential cases, and another research group blinded to the exposure status collected the outcome and follow-up of the case. The first and the corresponding authors were responsible for all data analyses and interpretation.Adjuvant chemotherapy was administered after surgery for NSCLC. Adjuvant chemotherapy dose intensity was applied according to the NCCN guidelines. During postoperative treatment, chemotherapy quickly induces myelosuppression, and rhG-CSFs were given to relieve or prevent myelosuppression. rhG-CSFs were administered 24 h after adjuvant chemotherapy. Patients were divided into two groups based on the use or non-use of rhG-CSF. All rhG-CSF users had at least one rhG-CSF prescription. The effects of the dose of rhG-CSF exposure on the postoperative metastasis risk of NSCLC were also analysed. Four subgroups  were divided based on the cumulative total rhG-CSF dose. Because of the impact of dose use over time, dose density of rhG-CSF on metastasis was used to evaluate the effects of density of rhG-CSF treatment on the outcome variable metastasis. Length of rhG-CSF exposure time (in days) was measured from the first prescription to the last to calculate the dose density of rhG-CSF treatment.We reviewed medical records to extract data on demographics, clinicopathological characteristics, and treatment histories: age, weight, sex, smoking habit (never smoked or smoked), anamnesis, histological type, grade, and pathological stage. During the postoperative chemotherapy and follow-up period, regimen and number of chemotherapies, the severity of myelosuppression (Common Terminology Criteria for Adverse Events (CTCAE), version 4.03), fever , and antibiotics were collected.All selected NSCLC patients achieved disease-free status after surgery. We reviewed clinical notes and checked all routine imaging records of enrolled patients after adjuvant chemotherapy, including ultrasound (US), thoracic radiography, computed tomography (CT), magnetic resonance imaging (MRI), and single-photon emission computed tomography (SPECT) at the time of diagnosis or during follow-ups, to assess their metastatic status. Metastasis-free survival (MFS) was defined as the duration from the day of chest surgery to investigator-assessed radiographic organ or node metastasis. MFS was censored on the last follow-up date if patients were alive and without any evidence of metastasis. Patient status of metastasis 1, 2, and 3 years after surgery was also measured. Patients without documented clinical or radiographic disease date of metastasis were censored at the last follow-up.χ2 tests for categorical data were used to analyse each covariate’s required data distribution between the rhG-CSF and No rhG-CSF groups. We used the Kaplan-Meier method to calculate overall MFS rates, and the differences in survival for univariate comparisons were calculated using the log-rank test. Forest plots were used to describe the underlying effect modification by each covariate. Interaction analysis was performed by calculating the respective categorical variable product terms individually in the model. Whether rhG-CSF and other covariates had an independent effect on NSCLC patients’ metastasis following postoperative chemotherapy separately was evaluated using univariate logistic regression and multivariate logistic regression models. The level of significance was established at p = 0.05. The statistical software packages R  and EmpowerStats  were used to perform all statistical analyses.Continuous variables are shown as the means ± SDs, and categorical variables are shown as frequencies (percentages). The t-test  or Kruskal-Wallis rank-sum test  for continuous variables and p < 0.001). Most patients with myelosuppression after chemotherapy tend to use rhG-CSF in clinical practice. A total of 107 patients received rhG-CSF as primary prophylaxis, and 139 patients received rhG-CSF as secondary prophylaxis. Essential characteristics were comparable between the No rhG-CSF and rhG-CSF groups. Patients in the two groups were similar in age, weight, sex, smoking, histology, differentiation, pathological stage, chemotherapy, fever, antibiotics, and complications. We excluded patients with a poor general physical condition before surgery and chemotherapy treatment. Therefore, the Eastern Cooperative Oncology Group performance status (ECOG PS) level of all postoperative NSCLC patients was 0–1, and there was no noticeable difference between the rhG-CSF and No rhG-CSF groups.Of the 307 postoperative NSCLC patients included in the study, 246 80.1%) patients who received rhG-CSF treatment during chemotherapy were used as the study group (rhG-CSF group), and 61 (19.9%) patients who did not receive rhG-CSF were included in the control group (No rhG-CSF group). The clinical characteristics and baseline parameters of the No rhG-CSF and rhG-CSF groups are presented in Table % patientp < 0.05). RhG-CSF treatment significantly correlated with metastasis in univariate Cox regression analysis . We found that the different doses and density gradients of rhG-CSF likely played an essential role in distant organ metastases , which is consistent with previous studies [A total of 136 of the 307 postoperative NSCLC patients (44.30%) had distant organ metastases following the chest surgery over the median follow-up period of 33.63 months. A total of 119 (48.3%) patients were in the rhG-CSF group, and 17 27.9%) patients were in the No rhG-CSF group, which suggests that the DOM rate was significantly higher in the study group than the control group (7.9% pati studies .Table 2Ep = 0.007), 2 years  and 3 years  of postoperative follow-up  and 1.33 times in the adjusted II model . We performed similar analyses for postoperative follow-up at 1, 2, and 3 years, and rhG-CSF treatment was associated with an increased risk of metastasis in the postoperative NSCLC patients  except for myelosuppression . Among patients who presented without myelosuppression after chemotherapy, rhG-CSF treatment resulted in a significantly high risk of metastasis . In contrast, rhG-CSF treatment reduced the risk of metastasis in patients who presented myelosuppression . Compared to the patients who had myelosuppression after chemotherapy, patients who presented without myelosuppression had a higher risk correlation of rhG-CSF use with metastasis , and at postoperative follow-up 1 year , 2 years  and 3 years  , in the non-adjusted and adjusted models (Table p for trend> 0.05). When the factor of time was considered, the dose density of rhG-CSF, which is dose of rhG-CSF used in the length of rhG-CSF exposure time, a positive relationship between the dose density of rhG-CSF treatment and risk of metastasis was observed for all models. Statistically significant results of trend tests were obtained in the total sample (p for trend< 0.001) (Table p for trend< 0.05)  Table . The ana5) Table .Distant organ metastasis is always associated with poor prognosis and shorter survival in NSCLC patients . The speThe influence of tumour staging on patient metastasis has been generally recognized . For patCSF was first discovered by Don Metcalf et al. . Mice trn = 125) group during cycle 1 of chemotherapy in NSCLC patients [n = 51) of patients with NSCLC who received chemotherapy without rhG-CSF treatment exhibited FN. There was a relatively low occurrence of FN in NSCLC patients even without rhG-CSF. It was evident that a large proportion of patients who received mild chemotherapy could also safely survive the risk period after chemotherapy in the absence of rhG-CSF. Guidelines pay close attention to whether the use of rhG-CSF reduces the occurrence of FN and its complications, the risk of infection and death, the duration of severe neutropenia, the duration of antibiotic use, the length of hospital stay, bone pain, and other indicators. However, it may ignore the dark side of rhG-CSF supplements, such as promoting cancer metastasis. The risk of metastasis with excessive use of rhG-CSF was increased remarkably in most of these patients, which led us to consider whether certain precautions, such as avoiding some infection factors and increasing nutrition after chemotherapy, could take the place of rhG-CSF use to lower the risk. Although patients with breast cancer benefitted from rhG-CSF use in extending DFS and OS, any damage from rhG-CSF use could be whitewashed by the profit supported by dose-intensity chemotherapy regimens [Current guidelines for rhG-CSF of National Comprehensive Cancer Network (NCCN) treatment recommend prophylaxis with rhG-CSF when the risk of FN is approximately 20% or higher or the risk of FN ranges from 10 to 20% combined with one or more risk factors, such as age and recent history of surgery . The purpatients  found thregimens . Howeverregimens .A developing number of studies –30 identNeutrophil extracellular traps (NETs) are a network structure consisting of modified chromatin structures and decorated with given cytoplasmic and granular proteins, which likely also played a crucial part in the enhanced risk of DOM with rhG-CSF use . NETs, fThe analyses of this research offer proof of the effect of rhG-CSF on the development of DOM in NSCLC patients with postoperative chemotherapy. Therefore, a physician in clinical practice should administer rhG-CSF to NSCLC patients based on chemotherapy regimens and the patient’s own risk factors using rhG-CSF once myelosuppression exists. The recommended guidelines for rhG-CSF use should formulate more comprehensive standards. Populations that are suitable to use rhG-CSF are also worthy of further exploration.Our study has the following limitations. First, due to loss to follow-up, we did not dissect the OS of participants in this study, which should be shown in subsequent research. Second, our study only included NSCLC patients who received chemotherapy postoperatively. Future studies should consider more tumour types. DOM was primarily judged using imaging examination and not biopsy, which likely caused false-positive results, despite the diagnosis estimated by two radiologists. Pathological discoveries may be used to supply evidence microenvironment variations in the metastatic organs. Lastly, this study was a retrospective study, and some selection biases may be present. Prospective research is also needed to verify these conclusion.Among NSCLS patients following postoperative chemotherapy, we found that rhG-CSF use was related to a high risk of metastasis, and this risk may increase significantly with the increase in dose density of rhG-CSF. The study suggests that patients without myelosuppression have more disadvantages from rhG-CSF. Our findings, if affirmed, lead to far-reaching clinical and public health influence because of the highest mortality rate of NSCLC globally. This result suggests that identifying patients with or without myelosuppression after chemotherapy could help guide the rational drug use of rhG-CSF and avoid its risks in clinical use.


The application of single-cell RNA sequencing (scRNA-seq) in biomedical research has advanced our understanding of the pathogenesis of disease and provided valuable insights into new diagnostic and therapeutic strategies. With the expansion of capacity for high-throughput scRNA-seq, including clinical samples, the analysis of these huge volumes of data has become a daunting prospect for researchers entering this field. Here, we review the workflow for typical scRNA-seq data analysis, covering raw data processing and quality control, basic data analysis applicable for almost all scRNA-seq data sets, and advanced data analysis that should be tailored to specific scientific questions. While summarizing the current methods for each analysis step, we also provide an online repository of software and wrapped-up scripts to support the implementation. Recommendations and caveats are pointed out for some specific analysis tasks and approaches. We hope this resource will be helpful to researchers engaging with scRNA-seq, in particular for emerging clinical applications.The online version contains supplementary material available at 10.1186/s40779-022-00434-8. Complex tissues consist of a variety of cell types that occur in a huge variety of mixtures and states. The functional genomic information contained within each cell is often quite different from the neighboring cell populations and even cells of the same type. This means that the molecular analyses of cell populations in bulk tissues are inherently unreliable and insensitive. The incredible sensitivity and specificity that can be achieved by quantifying molecular alterations at single-cell resolution have led to unprecedented opportunities for uncovering the molecular mechanisms underlying the pathogenesis and progression of the disease . Since iSince the emergence of commercial single-cell platforms, including those offered by 10 × genomics , 30 and https://github.com/WXlab-NJMU/scrna-recom. To accommodate upcoming advanced approaches and more application scenarios, we will keep the computational scripts updated.To overcome the barriers in scRNA-seq data analysis, in particular for biomedical studies, this review aims to: 1) summarize the recent advances in algorithm development and benchmarking results for every analysis task in analyzing biomedical scRNA-seq data, and 2) introduce a workflow comprised of recommended software tools that are more appropriate for biomedical applications. The workflow covers basic scRNA-seq data processing, quality control (QC), feature selection, dimensionality reduction, cell clustering and annotation, trajectory inference, cell–cell communications (CCC), transcription factor (TF) active prediction and metabolic analysis. Along with the recommended workflow, we also provide example computational scripts together with the software environment setting, which may facilitate researchers to conduct the data analysis locally. The computational code is available at Typical data analysis steps of scRNA-seq can be generally divided into three stages: raw data processing and QC, basic data analysis applicable for almost all scRNA-seq data sets, and advanced data analysis that should be tailored to specific research scenarios. While basic data analysis steps include data normalization and integration, feature selection, dimensionality reduction, cell clustering, cell type annotation and marker gene identification. The advanced data analysis tasks consist of trajectory inference, CCC analysis, regulon inference and TF activity prediction, and metabolic flux estimation.ScRNA-seq experiments need to be carefully designed to optimize the capability in addressing scientific questions . Before https://github.com/singleron-RD/CeleScope) for Singleron’s systems, alternative tools including UMI-tools . By contrast, the method SingleCellSignalR is based on the product of LR gene expression levels [To facilitate the above investigation, known ligand-receptor interactions (LRIs) have been manually curated and deposited in databases Fig. a. To datn levels .Fig. 3ThRecently, computational methods for predicting CCC based on scRNA-seq data have been continuously developed . The CCCThe emerging computational methods for identifying CCC have improved our understanding of the microenvironment for disease development. However, all the methods depend on prior knowledge of LRIs and statistical or machine learning models to predict potential CCC events. Alternatively choosing LRI resources and prediction approaches may result in different results, yet the impact of the choice on the results is largely unknown. To address this issue, one recent study systematically compared 16 resources and 7 methods for CCC inference, as well as the consensus of the compared methods . The comTFs play essential roles in gene expression regulation, and are involved in various physiological and pathological processes of humans . It has One important resource in recognizing regulons is the TF-target databases. The Animal Transcription Factor DataBase  , JASPAR Integrating single-cell gene expression and the comprehensive TF-target information, there have been many methods developed for inferring regulons and TF activity. Coexpression analysis, such as weighted gene co-expression network analysis (WGCNA) , has beeDespite many methods developed for gene regulation analysis based on scRNA-seq, a rigorous judgment on the inferred results needs to be made, due to the complexity of transcriptional regulation and the insufficient information provided by scRNA-seq data. Performing validation experiments may make the inferred results more solid , 254.Metabolism is at the core of all biological processes, and metabolic dysregulation is a hallmark of many diseases including cancer, diabetes, and cardiovascular disease . AlthougThe computational tools for scRNA-seq-based metabolic analysis can be classified into two major categories: pathway-based analysis and flux balance analysis (FBA)-based methods  Fig. 5)5). For tThe other category is the FBA-based methods, where constraint-based mathematical models are utilized to systematically simulate metabolism in reconstructed metabolic networks . The rechttps://github.com/WXlab-NJMU/scrna-recom. Notably, due to time and space constraints, we are unable to incorporate all popular tools into the analysis pipelines on the GitHub site; however, we provide a list of currently available tools with accessible links for users’ convenience  was necessary to process the scRNA-seq data for identifying Polylox transcript variants in lineage tracing [Focusing on single-cell transcriptomics, we have reviewed almost all respects of typical analysis of scRNA-seq data, ranging from QC, basic data processing, to high-level analysis including trajectory inference, CCC estimation and metabolic analysis. To facilitate researchers conducting the analysis on their data, we have constructed an online software/script repertoire for these analysis steps, and will keep it updated to cover more research scenarios. We also offer a step-by-step command line interface (CLI) for wrapping up the R and Python scripts for scRNA-seq analysis. The step-wise commands can be flexibly combined and tailored for specific applications due to the diversity on scientific questions and experimental design. Moreover, incorporating cutting-edge technologies, the analysis steps reviewed above may not cover every specifically required task. Indeed, additional analysis pipeline  in a cell subtype of interest might be misleading, although it is more intuitive. This is because massive cells are usually profiled in a scRNA-seq experiment, and consequently cell points can be buried by some others in the two-dimensional visualization. Other types of plots that directly and more quantitatively demonstrate the composition would be more suitable.Many other aspects of scRNA-seq data analysis are advancing rapidly. ScAPAtrap , Sierra In addition to gene expression regulation by TFs, trans-factors like RNA binding proteins (RBPs) and microRNAs typically bind to the 3’UTR of genes to modulate RNA stability, which also contributes to cellular RNA concentration. Based on collections of RBP and microRNA target genes , 282, RBVery recently, Live-seq has been developed to convert scRNA-seq from an end-point type assay to a temporal analysis workflow, by keeping cells alive while extracting RNA from individual cells . It is aAdditional file 1. Table S1: Tools for analyzing single-cell RNA-seq data, with references and links.


Background and objectives: Traditional Chinese medicines (TCMs) are widely prescribed to relieve ischemic heart disease (IHD); however, no cohort studies have been conducted on the use of TCMs for patients with IHD. The aim of the study was to analyze TCM prescription patterns for patients with IHD. Materials and Methods: The retrospective population-based study employed a randomly sampled cohort of 4317 subjects who visited TCM clinics. Data were obtained from the National Health Insurance Research Database (NHIRD) of Taiwan for the period covering 2000 to 2017. Data analysis focused on the top ten most commonly prescribed formulae and single TCMs. We also examined the most common two- and three-drug combinations of TCM in single prescriptions. Demographic characteristics included age and sex distributions. Analysis was performed on 22,441 prescriptions. Results: The majority of TCM patients were male (53.6%) and over 50 years of age (65.1%). Zhi-Gan-Cao-Tang (24.76%) was the most frequently prescribed formulae, and Danshen (28.89%) was the most frequently prescribed single TCM for the treatment of IHD. The most common two- and three-drug TCM combinations were Xue-Fu-Zhu-Yu-Tang and Danshen” (7.51%) and “Zhi-Gan-Cao-Tang, Yang-Xin-Tang, and Gua-Lou-Xie-Bai-Ban-Xia-Tang” (2.79%). Conclusions: Our results suggest that most of the frequently prescribed TCMs for IHD were Qi toning agents that deal with cardiovascular disease through the promotion of blood circulation. The widespread use of these drugs warrants large-scale, randomized clinical trials to investigate their effectiveness and safety. Ischemic heart disease (IHD), also known as coronary heart disease, is a leading cause of death and disability worldwide. According to global statistics, the total number of IHD cases reached 182 million in 2019, resulting in roughly 9 million deaths .κB and HIF-1α signaling .In Taiwan, TCM doctors make diagnoses in accordance with ICD-9-CM coding. In the current study, we focused exclusively on patient data with the diagnosis code for IHD. We then sorted the 4317 subjects who visited TCM clinics for IHD. This led to the retrieval of 22,441 prescriptions for TCM. Our database included six interlinked variables: month and year of the fee, application type, hospital code for hospital identification, application date, case type, and serial number. Based on the theory of Chinese medicine, there are four types of TCM prescriptions: (1) a single herb; (2) a composition of multiple herbs with various dosages (Fu-Fang); (3) a regimen of herbs with fixed dosages in line with classical or well-known TCM textbooks (Fang-Ji); and (4) a classical formula in which a drug compound is combined with herbs (Chia-Chien-Fang). Note that combinations of ingredients are in powder form, which facilitates mixing within a single prescription.All IHD subjects were defined by the diagnosis code ICD9 410–414 (ICD10: I20–I25). Age and gender-specific frequencies for the use of TCMs among patients with IHD from 2000–2017 were examined. We used SAS 9.4  to link ambulatory care claim data and data with the details of ambulatory prescriptions. Frequency and patterns of TCM formula or single herbs were performed by statistical software R, which is a publicly available software in our institution.Association rules and frequent itemset mining are well-established approaches to discovering specific relationships among variables in large databases. Data mining was first developed in the 19th century ,8. In thUsing the screening diagnosis code ICD9 410–414 (ICD10: I20–I25), we identified 4317 patients who sought TCM treatment for IHD. The patients were distributed according to age into four categories: ≤34, 35–49, 50–64, and ≥65 years. Gender classification was based exclusively on physical gender . As shown in As shown in As shown in Ten TCMs (singly or in formulas) accounted for roughly 15% of the prescriptions. Most single prescriptions contained 5 to 7 items (average of 6.15) . The mosIn ancient TCM texts, the terms used to describe diseases related to IHD included “Xin Tong” (heartache), “Xin Jue” (heart syncope), “Zhu Xin Tong” (acute heartache), and “Zhen Xin Tong” (true heartache). In the oldest ancient pharmacopeia “Huangdi Neijing” , there are detailed descriptions of IHD: “For the true heartache, if your hands and feet become blue and reach your elbows and knees, the pain is severe. It usually occurs in the morning and fades in the evening, or it occurs in the evening and fades in the morning”. These descriptions of pain in the precordial area were attributed to poor qi and blood flow. In some cases, they mentioned pain in the shoulders and back, shortness of breath and wheezing, and an inability to lie down, all of which fell within the designation of “heartache”. These descriptions are similar to the clinical manifestations of coronary heart disease and angina pectoris.In Western medicine, most diagnoses of coronary heart disease are based on the results of angiography or computer tomography. Due to the high risk of mortality, treatment often involves highly invasive emergency procedures. TCM is meant to prevent coronary artery obstruction and disease using non-invasive methods . This maSalvia miltiorrhiza), which possesses a bitter taste and cold nature associated with the heart and liver meridians. It was classified as a top-grade medication in “Shennong Bencao Jing”  for the treatment of heart and spleen disease, to clear the blood circulation system, relieve irritation, and replenish the Qi. Danshen has also been recommended for the treatment of angina pectoris in “Yi Xue Xiao Jin Zhen” . A number of recent scientific studies have examined the cardioprotective effects of Danshen  [The most frequently prescribed single TCM was Danshen (dried root of ll form) . The conll form) ,44,45,46ll form) ,48,49,50Curcumae Longae Radix), which is the dry root of Zingiberaceae cultivars, such as Curcuma wenyujin, Curcuma kwangsiensis, Curcuma longa, and Curcuma phaeocaulis. Yujin also possesses a bitter taste and cold nature associated with the heart, gut, and liver meridians. According to the “Bencao Beiyao” , Yujin can cool the heart and blood, disperse stagnation, and normalize the gallbladder to cure jaundice. Clinically, it is often used to treat chest, flank, and abdominal pain induced by Qi stagnation and blood stasis. A number of recent scientific studies have reported that Yujin activates blood flow and mediates blood stasis by inhibiting the aggregation of platelets, slowing the over-activation of the coagulation, and promoting blood circulation [Another frequently prescribed single TCM was Yujin  recommended the use of Yanhusuo “to treat dying heartache”. The “Bencao Gangmu”  also reported on the therapeutic effects of Yanhusuo in treating blood and qi stagnation as well as pains throughout the body. Recent scientific studies have extracted alkaloids, such as dextrocorydine, tetrahydropalmatine, and protopine, which have been shown to have potent analgesic and antispasmodic effects on the central nervous system. Another alakaloid, l-Tetrahydropalmatine, exhibited cardioprotective effects against acute global cerebral ischemia-reperfusion injury by activating the PI3K/Akt/eNOS/NO pathway and decreasing the accumulation of inflammatory factors, such as TNF-α and MPO [Yanhusuo is the dry tuber of  and MPO . Dehydro and MPO .Gegen possesses a flat, sweet taste and has been associated with the Yangming meridian and spleen meridian. Traditionally, it was used to uplift the qi in the stomach, enhance fluid production, and quench the thirst as well as promote sweating and relieve muscles tightness and fever. It has traditionally been used to treat spleen and stomach diseases as well as diarrhea. Recent scientific studies have demonstrated that Gegen can dilate coronary and cerebral blood vessels and increase blood flow. Gegen contains bioactive ingredients  and other flavones and isoflavones, which have been shown to inhibit platelet aggregation and improve the metabolism of the ischemic myocardium ,54. A nuGlycyrrhizae and Radix Ginseng, glycyrrhizin and ginsenosides, all of which have been shown to improve the myocardial contractility and cardiac function by inhibiting of Na+/K+−ATPase [The most popular TCM formula in this study was Zhi-Gan-Cao-Tang, which can be traced back to the ancient pharmacopeia “Shanghan Lun” . It has been recommended for the treatment of typhoid fever, pulse congestion, and heart palpitations, owing to the effects on replenishing qi, nourishing yin, and tonifying yang as well as rejuvenating the pulse. The main constituents are from Radix +−ATPase . They ha+−ATPase ,31.Bupleurum chinense DC./Bupleurum scorzonerifolium Willd.), Chuanxiong (Ligusticum chuanxiong Hort.), and Chishao  have been linked to analgesic effects, while Chuanxiong, Chishao, Zhishi (Citrus aurantium L.), and Gancao  have been linked to the alleviation of smooth muscle spasms. Xue-Fu-Zhu-Yu-Tang has been shown to reduce the concentration of triglycerides in serum, decrease the ratio of TXA2/PGI2, and decrease the production of proinflammatory cytokines in rats fed a high cholesterol diet [The second most popular TCM formula in this study was Xue-Fu-Zhu-Yu-Tang, which can be traced back to the pharmacopeia “Yilin Gaicuo” . This combination of 11 herbs is used mainly to promote blood circulation and remove blood stasis or as a supplement to soothe the liver or regulate the qi. Clinical applications include the treatment of blood stasis diseases, such as rheumatic heart disease, coronary heart disease, and angina pectoris, as well as chest contusion, intercostal neuralgia, chest pain of costochondritis, and sequelae of concussion. Pharmacological analysis has revealed that Xue-Fu-Zhu-Yu-Tang has anticoagulant and thrombolytic effects, including the dilation of peripheral blood vessels and coronary arteries to promote blood circulation . In bloorol diet . Xue-Fu-rol diet . In a swrol diet .Another famous TCM formula, Sheng-Mai-Yin, can be traced back to the ancient pharmacopeia “Lantai Gueifan” . This formula, containing Panax ginseng, Ophiopogon japonicus, and Schisandra chinensis, is commonly used to deal with coronary heart disease, angina pectoris, viral myocarditis, heart failure, and other heart diseases. Recent scientific studies have reported that it can also have protective effects on the heart, such as reducing early ventricular contraction and paroxysmal tachycardia, by suppressing the sodium and potassium ATPase pumps of the myocardial cell membrane . InjectiPinellia ternata (Thunb.) Makino) is frequently prescribed for IHD. This formula contains the cold bitter Gualouren (Trichosanthes kirilowii Maxim./Trichosanthes rosthornii Harms), which is used to reduce phlegm and relieve chest tightness. It also contains the warm smooth Xiebai (Allium macrostemon Bunge), which is used to promote yang qi and activate the blood. Another constituent, Banxia, is used to reduce dampness, limit phlegm production, and halt vomiting. Gua-Lou-Xie-Bai-Ban-Xia-Tang is meant for coronary heart disease involving qi deficiency, phlegm, and blood stasis and is clinically used for chest obstructive syndrome, such as coronary heart disease angina, intercostal neuralgia, arrhythmia, chest soft tissue injury, intercostal chondritis, and emphysema. Recent scientific evidence suggests that Gua-Lou-Xie-Bai-Ban-Xia-Tang could have cardioprotective effects in ischemia-reperfusion models, including the inhibition of NF-κB and inflammatory cytokines [The TCM formula Gua-Lou-Xie-Bai-Ban-Xia-Tang can be traced back to the pharmacopeia “Jingui Yaolue” . This modification of Gua-Lou-Xie-Bai-Bai-Jiou-Tang through the addition of Banxia , or Western drugs (such as Atorvastatin) in the treatment of cardiovascular diseases [This study was prone to inevitable errors, including inaccurate diagnoses. In many cases, patients with chest or heart discomfort receive a diagnosis of IHD (ICD9 diagnosis code: 410–414 and ICD10 diagnosis code: I20–I25) before undergoing a thorough examination. Or, for patients with IHD, only the other forms of heart disease (ICD9-CM diagnosis code 420–429 and ICD10 diagnosis code: I30–I52), chest pain (ICD9-CM diagnosis code 786.5 and ICD10 diagnosis code: R079), and other chest pain (ICD9-CM diagnosis code 786.59 and ICD10 diagnosis code: R0789) were declared in the NHI database, which also caused the error of case number reducing. Another source of error was the stacking of formulas with single TCMs, which increased the proportion of specific herbs. These errors are difficult to eliminate; however, they do not reduce the credibility of the research, due to the fact that these prescriptions were deemed effective by doctors in terms of medical theory and empirical use. In addition, this study was subject to a number of limitations. Identification of the best TCM formula for a specific patient should be based on careful analysis of all clinical information obtained through the four diagnostic methods  and focusing on the eight principal syndromes . We were unable to collect detailed medical records for each patient, and many IHD patients also underwent modern medical treatments, such that the efficacy of the TCMs could not be assessed objectively. Note also that in Taiwan, many Chinese herbal medicines or health foods containing Chinese herbal ingredients can be purchased directly from TCM pharmacies or drugstores, which makes it impossible to assess the effects of inadvertent drug interactions. For example, the combined use of Warfarin and herbal medicines, such as Danshen and Licorice (the major herb in Zhi-Gan-Cao-Tang), can over-activate the anticoagulant effect and lead to bleeding complications . Danshendiseases .Salvia miltiorrhiza), and the most popular formula was Zhi-Gan-Cao-Tang. The safety and therapeutic effects of these TCMs will require further exploration in future studies.This paper reports on TCM prescription patterns for IHD from the NHI Research Database. The single herb most frequently prescribed for IHD was Danshen (


Functional somatic disorders (FSDs) are characterized by persistent and disabling physical symptoms that cannot be attributed to well-defined somatic disorders. In adolescents, the prevalence is around 4-10%. Evidence from adult populations suggests that cortisol plays a role in the development and perpetuation of FSDs, but little is known regarding adolescents. As cortisol accumulates in hair over time, hair cortisol concentration (HCC) is a promising new biomarker for long-term physiological stress. Moreover, adult studies have found associations between HCC levels and self-perceived stress.To compare HCC levels between adolescents with severe FSDs and adolescents from the general population. Furthermore, to investigate the association between HCC and self-perceived stress.The data are retrieved from two projects: the AHEAD trial, including 91 15-19-year-old adolescents diagnosed with a severe FSD, and the Copenhagen Child Cohort 2000 (CCC2000), including data on 1455 16-17-year-old adolescents. Hair samples were collected for HCC analysis, and web-based questionnaires were used to asses self-perceived stress. Functional somatic symptoms were assessed with the Bodily Distress Syndrome (BDS) checklist.The data have been collected and will be analysed and presented at the congress.This study can contribute with knowledge about the potential role of cortisol in FSDs in adolescents, and whether self-perceived stress can be used as a marker for physiological stress measured by HCC. Treatments for adolescents with FSDs still need to be improved. The current study may help to understand whether future treatment strategies should include a greater focus on stress management.No significant relationships.