Integrated causal inference modeling uncovers novel causal factors and potential therapeutic targets of Qingjin Yiqi granules for chronic fatigue syndrome

Objective:Chronic fatigue syndrome(CFS)is a prevalent symptom of post-coronavirus disease 2019(COVID-19)and is associated with unclear disease mechanisms.The herbal medicine Qingjin Yiqi granules(QJYQ)constitute a clinically approved formula for treating post-COVID-19;however,its potential as a drug target for treating CFS remains largely unknown.This study aimed to identify novel causal factors for CFS and elucidate the potential targets and pharmacological mechanisms of action of QJYQ in treating CFS.Methods:This prospective cohort analysis included 4,212 adults aged≥65 years who were followed up for 7 years with 435 incident CFS cases.Causal modeling and multivariate logistic regression analysis were performed to identify the potential causal determinants of CFS.A proteome-wide,two-sample Mendelian randomization(MR)analysis was employed to explore the proteins associated with the identified causal factors of CFS,which may serve as potential drug targets.Furthermore,we performed a virtual screening analysis to assess the binding affinity between the bioactive compounds in QJYQ and CFS-associated proteins.Results:Among 4,212 participants(47.5%men)with a median age of 69 years(interquartile range:69–70 years)enrolled in 2004,435 developed CFS by 2011.Causal graph analysis with multivariate logistic regression identified frequent cough(odds ratio:1.74,95%confidence interval[CI]:1.15–2.63)and insomnia(odds ratio:2.59,95%CI:1.77–3.79)as novel causal factors of CFS.Proteome-wide MR analysis revealed that the upregulation of endothelial cell-selective adhesion molecule(ESAM)was causally linked to both chronic cough(odds ratio:1.019,95%CI:1.012–1.026,P=2.75 e^(−05))and insomnia(odds ratio:1.015,95%CI:1.008–1.022,P=4.40 e^(−08))in CFS.The major bioactive compounds of QJYQ,ginsenoside Rb2(docking score:−6.03)and RG4(docking score:−6.15),bound to ESAM with high affinity based on virtual screening.Conclusions:Our integrated analytical framework combining epidemiological,genetic,and in silico data provides a novel strategy for elucidating complex disease mechanisms,such as CFS,and informing models of action of traditional Chinese medicines,such as QJYQ.Further validation in animal models is warranted to confirm the potential pharmacological effects of QJYQ on ESAM and as a treatment for CFS.

Effects of Intermittent Hypoxia Exposure on Symptoms of Chronic Fatigue Syndrome in Repeated Restraint Stress and Forced Swimming Induced-Mouse Model

Background: Chronic fatigue syndrome (CFS) shows as its main symptoms debilitating fatigue that is not relieved by physiological rest, depression, inflammation, learning disability and memory impairment. But, intermittent hypoxia, consisting of alternating exposure to hypoxia and normoxia, plays a very important role in improving CFS. However, the essential components for improving learning and memory in CFS patients as well as their mechanism are largely unknown. Objectives: This study aims to analyze the effects of 12% and 15% hypoxia on the expression of alpha tumor necrosis factor (TNF-α) and nuclear factor kappa B (NF-κB) in CFS induced-mouse model for clarifying the effects on the learning and memory function. Methods: A total of 48 type IC mice were used. The CFS mouse model was established using restrained stress and repeated forced swimming. Treatment of CFS was done by exposing CFS mice to intermittent hypoxia at 12% and 15%. The effects of intermittent hypoxia on learning and memory as well as its mechanism of action on inflammation were tested respectively with the Morris test, the SDS page, the immunohistochemistry technique and the Nissl staining. Results: We found that 12% and 15% intermittent hypoxia exposure improved learning capacity and memory of CFS induced-mice. SDS page showed that CFS caused higher TNF-α expression. By exposing CFS mice to 12% and 15% intermittent hypoxia, TNF-α expression decreased significantly, with a much better effect at 15%. Both TNF-α and NF-κB increased in CFS state and decreased after treatment with intermittent hypoxia. Conclusion: Intermittent hypoxia improves learning capacity and memory. It acted by decreasing NF-κB come to down-regulating TNF-α and ameliorates learning capacity and memory impairment in CFS mice.

Effects of Exercise on Behavior and Peripheral Blood Lymphocyte Apoptosis in a Rat Model of Chronic Fatigue Syndrome

This study examined the effects of exercise on behavior and peripheral blood leukocyte apoptosis in a rat model of chronic fatigue syndrome （CFS）. Thirty-six healthy male Sprague-Dawley rats were equally randomized into 3 groups： the control group, CFS model group and the exercise group in terms of body weight. A total of 25 rats entered the final statistical analysis due to 11 deaths during the study. CFS model was established by subjecting the rats in CFS model group and exercise group to electric shock, chronic restraint stress and cold water swim. Besides, rats in the exercise group took rtmning wheel exercise. After a week of conditioning feeding, model construction and running wheel exercise were performed simultaneously, and lasted for 23 consecutive days. The behavior experiments, including running wheel exercise, open-field test, tail suspension test and Morris water maze test, were conducted, either before or after the model establishment. Rats were sacrificed and peripheral blood was obtained for the assessment of lymphocyte apoptosis index by flow cytometry （FCM）. It was found that as compared with those in the control group, the weight of the rats was decreased obviously （P〈0.01）, the mobility time in the open-field and the tail suspension tests was shortened significantly （P〈0.01）, the time to locate the platform was enhanced （P〈0.01） and the cell apoptosis index was increased substantially （P〈0.01） in the CSF model group. Meanwhile, in comparison to the model group, the behavior in the open-field and the tail suspension tests was improved significantly （P〈0.05）, and the apoptosis index decreased remarkably （P〈0.01） in the exercise group. It is concluded that sport intervention can prevent lymphocyte apoptosis and improve animal behavior rather than the memory.

CLINICAL OBSERVATION ON THE TREATMENT OF CHRONIC FATIGUE SYNDROME WITH DEEPLY PUNCTURED ELONGATED NEEDLE COMBINED WITH COMPREHENSIVE NURSING

Objective： To observe the therapeutic effect of deep puncture of Zusanli （足三里 ST 36） and Qihai （气海 CV 6） with elongated needle combined with comprehensive nursing in the treatment of chronic fatigue syndrome （CFS） so as to provide new way and effective method for CFS. Methods： A total of 100 cases of CFS patients were evenly randomized into treatment group and control group which were treated with elongated needle puncture of ST 36 and CV 6 plus nursing and oral ackninistration of Shiquan Dobu Tong （十全大补汤 Decoction of Ten Powerful Tonics） separately. Acupuncture and medication were given once daily, with 7 sessions being a therapeutic course, 4 courses altogether. Results： After 4 courses of treatment, of the two 50 cases in treatment and control groups, 4 （8.0%） and 2 （4.0%） were cured, 37 （74.0%） and 16 （32.0%） experienced marked im- provernent, 6 （ 12.0 % ） and 19 （38.0 % ） had improvement, 3 （6.0 % ） and 13 （26.0 % ） tailed, with the total effective rates being 94.0% （47/50） and 74.0% （37/50） respectively. The total effective rate and markedly effective rate of treatment group were significantly higher than those of control group （P〈0.05, 0.01）. Conclusion; Deep puncture of Zusenli （ST 36） and Qihai （CV 6） with elongated needle combined with comprehensive nursing has a definite therapeutic effect in the treatment of chronic fatigue syndrome.

Accurate diagnosis of myalgic encephalomyelitis and chronic fatigue syndrome based upon objective test methods for characteristic symptoms

Although myalgic encephalomyelitis(ME) and chronic fatigue syndrome(CFS) are considered to be synonymous,the definitional criteria for ME and CFS define two distinct,partially overlapping,clinical entities.ME,whether defined by the original criteria or by the recently proposed criteria,is not equivalent to CFS,let alone a severe variant of incapacitating chronic fatigue.Distinctive features of ME are:muscle weaknessand easy muscle fatigability,cognitive impairment,circulatory deficits,a marked variability of the symptoms in presence and severity,but above all,post-exertional "malaise":a(delayed) prolonged aggravation of symptoms after a minor exertion.In contrast,CFS is primarily defined by(unexplained) chronic fatigue,which should be accompanied by four out of a list of 8 symptoms,e.g.,headaches.Due to the subjective nature of several symptoms of ME and CFS,researchers and clinicians have questioned the physiological origin of these symptoms and qualified ME and CFS as functional somatic syndromes.However,various characteristic symptoms,e.g.,post-exertional "malaise" and muscle weakness,can be assessed objectively using wellaccepted methods,e.g.,cardiopulmonary exercise tests and cognitive tests.The objective measures acquired by these methods should be used to accurately diagnose patients,to evaluate the severity and impact of the illness objectively and to assess the positive and negative effects of proposed therapies impartially.

Induced pluripotent stem cells as suitable sensors for fibromyalgia and myalgic encephalomyelitis/chronic fatigue syndrome

BACKGROUND Fibromyalgia(FM)and myalgic encephalomyelitis/chronic fatigue syndrome(ME/CFS)are devastating metabolic neuroimmune diseases that are difficult to diagnose because of the presence of numerous symptoms and a lack of specific biomarkers.Despite patient heterogeneity linked to patient subgroups and variation in disease severity,anomalies are found in the blood and plasma of these patients when compared with healthy control groups.The seeming specificity of these“plasma factors”,as recently reported by Ron Davis and his group at Stanford University,CA,United States,and observations by our group,have led to the proposal that induced pluripotent stem cells(iPSCs)may be used as metabolic sensors for FM and ME/CFS,a hypothesis that is the basis for this indepth review.AIM To identify metabolic signatures in FM and/or ME/CFS supporting the existence of disease-associated plasma factors to be sensed by iPSCs.METHODS A PRISMA(Preferred Reported Items for Systematic Reviews and Meta-analysis)-based systematic review of the literature was used to select original studies evaluating the metabolite profiles of FM and ME/CFS body fluids.The MeSH terms“metabolomic”or“metabolites”in combination with FM and ME/CFS disease terms were screened against the PubMed database.Only original studies applying omics technologies,published in English,were included.The data obtained were tabulated according to the disease and type of body fluid analyzed.Coincidences across studies were searched and P-values reported by the original studies were gathered to document significant differences found in the disease groups.RESULTS Eighteen previous studies show that some metabolites are commonly altered in ME/CFS and FM body fluids.In vitro cell-based assays have the potential to be developed as screening platforms,providing evidence for the existence of factors in patient body fluids capable of altering morphology,differentiation state and/or growth patterns.Moreover,they can be further developed using approaches aimed at blocking or reversing the effects of specific plasma/serum factors seen in patients.The documented high sensitivity and effective responses of iPSCs to environmental cues suggests that these pluripotent cells could form robust,reproducible reporter systems of metabolic diseases,including ME/CFS and FM.Furthermore,culturing iPSCs,or their mesenchymal stem cell counterparts,in patient-conditioned medium may provide valuable information to predict individual outcomes to stem-cell therapy in the context of precision medicine studies.CONCLUSION This opinion review explains our hypothesis that iPSCs could be developed as a screening platform to provide evidence of a metabolic imbalance in FM and ME/CFS.

A Comparative Study of Antifatigue Effects of Taurine and Vitamin C on Chronic Fatigue Syndrome

Vitamin C and taurine (TR) are well known as active components for fatigue recovery. However, the mechanism of the anti-fatigue effects of vitamin C and TR is still unclear. Our study was designed to evaluate the anti-fatigue activities of vitamin C and TR in an animal test for fatigue and to compare the activities between vitamin C and TR. Materials and Methods: Vitamin C, TR or their combination were orally administrated to mice once daily for 15 days, and then metabolic activities such as blood glucose, triglyceride (TG), lactate, and lactate dehydrogenase (LDH) as well as antioxidant activities such as malondialdehyde (MDA) and superoxide dismutase (SOD) were determined (evaluated) after forced swimming test (FST). Results: Compared with the control group, C100, C200, and T50 showed a tendency to decrease mobility in FST. Moreover, TG (C100, C200, T200), LDH (C200), lactic acid (C100) and MDA (C50, C100, C200) levels were inhibited by vitamin C and TR. Conclusions: These results suggest that vitamin C and TR have anti-fatigue activities in mice, with vitamin C providing a stronger effect.

Effect of intradermal needle at five-zang Back-Shu points on treatment of chronic fatigue syndrome

Objective:To observe the effect of intradermal needle therapy on back-shu points of five zang organs in treating chronic fatigue syndrome(CFS)and provide evidence for the usage of unilateral back-shu point.Methods:Patients were randomly divided into bilateral back-shu point group,unilateral back-shu point group and non-acupoint group,remove shedding cases,each group remaining 30 patients.In bilateral back-shu point group,both sides of back-shu points of five zang organs were selected.In unilateral back-shu point group,only one side was selected,left or right alternately.The location where non-acupoint group inserted by intradermal needles were outward from the back-shu points,left or right alternately.Fatigue Scale-14(FS-14)was used to evaluate the fatigue level and the scores’improvement of physical fatigue and mental fatigue revealed the therapeutic effect on body constituents and spirits of CFS patient.Results:After the 8-week intradermal needle therapy,the total effective rate was 83.33%in the bilateral back-shu point group,which was 76.67%in the unilateral back-shu point group and 50.00%in the non-acupoint group.The curative effects of the two groups whose back-shu points of five zang organs were inserted by intradermal needles were better than the non-acupoint group(P<0.05).There was no significant difference in clinical efficacy between the bilateral back-shu point group and the unilateral back-shu point group(P>0.05).There were statistical differences in FS-14 questionnaires including total,physical and mental scores between before and after the treatment in both the two back-shu point groups(P<0.05).Only the scores of total and physical fatigue in decreased in non-acupoint group(P<0.05),which means it couldn’t help to alleviate mental fatigue statistically(P>0.05).Conclusions:(1)The intradermal needle therapy on back-shu points of five zang organs for CFS has a satisfying clinical efficacy in curing tiredness.It can help to reduce both physical and mental fatigue for the patients.Hence,it can regulate both body constituents and spirits simultaneously.(2)Needling the unilateral acupoints can get similar clinical efficacy to the bilateral points.It reveals that we can cut cost by reducing the consumption of the amount of intradermal needles used on one side.(3)Both the groups of back-shu points get better clinical efficacy than the non-acupoint group dramatically.The reason of why the intradermal needle can treat CFS maybe that back-shu points of five zang organs can regulate the functions of five zang organs characteristically.

TREATMENT OF TEN CASES OF CHRONIC FATIGUE SYNDROME WITH CHINESE HERBS,ACUPUNCTURE, AND DIET PRECAUTIONS

Ten cases of chronic fatigue syndrome (CFS) were treated for 4 weeks by acupuncture, Chinese herbs and diet precautions. Diagnosis of CFS was determined by conventional medicine criteria(CDC criteria for CFS), physical examination, and routine laboratory tests. TCM diagnosis of CFS relied on Four Examination Methods and Eight Principal Patterns. They were diagnosed as Ganyu Qizhi (liver and qi stagnation) and Pixu Qiruo (spleen and qi deficiency). The treatment method is to disperse the liver and facilitate qi, rehabilitate the spleen and strengthen qi (Shugan Liqi, Jianpi Yiqi). The result of the treatment indicated that the total effective rate was 100% in terms of fatigue, difficulty concentrating, headache, sore throat, muscle pain, joint aches, feverishness, rapid pulse, insomnia, depression, irregular menses, abdominal cramps, and night sweats. The percentages of specific symptoms of CFS in the patients were comparable to the conventional medical record (Table 1).

An Etiological Model for Myalgic Encephalomyelitis/Chronic Fatigue Syndrome

Kindling might represent a heuristic model for understanding the etiology of Myalgic Encephalomyelitis/chronic fatigue syndrome (ME/CFS). Kindling occurs when an organism is exposed repeatedly to an initially sub-threshold stimulus re-sulting in hypersensitivity and spontaneous seizure-like activity. Among patients with ME/CFS, chronically repeated low-intensity stimulation due to an infectious illness might cause kindling of the limbic-hypothalamic-pituitary axis. Kindling might also occur by high-intensity stimulation (e.g., brain trauma) of the limbic-hypothalamic-pituitary axis. Once this system is charged or kindled, it can sustain a high level of arousal with little or no external stimulus and eventually this could lead to hypocortisolism. Seizure activity may spread to adjacent structures of the limbic-hypothalamic-pituitary axis in the brain, which might be responsible for the varied symptoms that occur among patients with ME/CFS. In addition, kindling may also be responsible for high levels of oxidative stress, which has been found in patients with ME/CFS.

Chronic fatigue syndrome with history of severe infection combined altered blood oxidant status, and reduced potassium efflux and muscle excitability at exercise

It is documented that chronic fatigue syndrome (CFS) combines enhanced oxidative stress with altered muscle excitability. We hypothesized that these disorders may be accentuated when severe infection preceded the CFS symptoms. This case-control study compared 55 CFS patients to a matched control group of 40 healthy subjects. In twenty-five CFS patients, severe infection was reported within the three to seven?month period preceding the CFS symptoms. The others had practiced sport at high level. Plasma concentrations of potassium, a marker of lipid peroxidation (thiobarbituric acid reactive substances, TBARS), and an endogenous antioxidant (reduced ascorbic acid, RAA) were measured. Action potential (M-wave) was evoked in the vastus lateralis muscle to explore the muscle membrane excitability. All subjects performed a maximal incremental cycling exercise. Compared to control subjects, all CFS patients presented an elevated resting TBARS level and, during and after exercise, an altered M-wave configuration. History of infection was associated with marked significant increase in resting TBARS level, enhanced M-wave alterations, and also reduced exercise-induced potassium efflux. The magnitude of exercise-induced M-wave alterations was proportional to the baseline TBARS level. Severe infection preceding CFS seems to constitute a stressor inducing altered blood oxidant status and a reduced muscle excitability at work.

Subgroups of chronic fatigue syndrome based on psychiatric disorder onset and current psychiatric status

Few studies have examined the effects of psy-chiatric disorders occurring over a long dura-tion among patients with chronic fatigue syn-drome (CFS). The role of premorbid and current psychiatric disorders in impairment was ex-plored with a sample of 113 participants with CFS. Subgroups were created based on past and current psychiatric status including those whose psychiatric history was premorbid and current, postmorbid and current, past but not current, and those with no lifetime diagnosis. Results from a one-way MANOVA revealed that patients with a premorbid and current psychiat-ric disorder reported significantly higher pain severity, more somatic symptoms, poorer sleep quality, and poorer quality of life than those with no psychiatric history. Levels of fatigue and physical functioning among patients with CFS were unrelated to the four subgroups in this study. Although those with a premorbid and current psychiatric disorder were differentiated from those with no psychiatric history on some markers of impairment, the sample as a whole had severe fatigue-related impairment, which is the cardinal symptom of CFS. Implications for research are discussed.

Chronic fatigue syndrome: An update for psychiatrists

Chronic fatigue syndrome (CFS) is a poorly understood condition primarily characterized by debilitateing, persistent or recurrent fatigue, increased physical and mental fatigability, cognitive impairment and widespread musculoskeletal pain. During the past two decades, there have been heated debates about CFS among researchers, practitioners and patients. The existence of the disorder has been questioned, its underlying pathophysiology debated and an effective treatment opposed (such as antidepressants, stimulants or antibiotics). A lot of multidisciplinary literature is found about CFS, but to date, many psychiatrists seem to unknown the existence of this illness or think that it is a purely psychological disorder. However, CFS is sitting on the border between medicine and psychiatry. The aim of this review is to make psychiatrists aware of the existence of CFS and that they will, one day, be confronted with the management of this illness. Thus, this update allows understanding what is CFS, the diversity of physiopathology underlined and its management.

A Comparison of Cytokine Profiles of Chronic Fatigue Syndrome/Myalgic Encephalomyelitis and Multiple Sclerosis Patients

Background: Chronic Fatigue Syndrome, also known as Myalgic Encephalomyelitis (CFS/ME), is a debilitating condition that presents with a range of symptoms, including fatigue, cognitive dysfunction, muscular and joint pain, and may be immune-mediated. In particular, patients exhibit abnormal cytokine expression. Similarly, in Multiple Sclerosis (MS), patients display neuroimmunological symptoms, and abnormal cytokine expression, with some overlap in symptomology with CFS/ME. The purpose of this study was to compare Th1, Th2, Th17 cytokines, inflammatory cytokines and chemokines, in healthy controls, CFS/ME and MS patients. Methods: Serum samples were collected from healthy controls (n = 16, mean age = 50 ± 11.85 years), CFS/ME patients (n = 16, mean age = 49.88 ± 9.54 years) and MS patients (n = 11, mean age = 52.75 ± 12.81 years). The concentrations of 27 cytokines (IFN-γ, TNF-α, IL-12, IL-2, IL-1β, IL-4, IL-6, IL-10, IL-13, IL-5, IL-17, IL-1ra, IL-7, IL-8, IL-9, eotaxin, IP-10, MCP-1, MIP1α, MIP1β, PDGF-bb, RANTES, basic FGF, GCSF, GMCSF, VEGF and IL-15) were measured using a Bio-Plex Pro&#8482 kit. Results: IFN-γ, IL-10 and IL-5 were significantly higher in the serum of both CFS/ME and MS patients compared to the healthy controls (p ≤ 0.041). However, only the MS patients had significantly elevated levels of IL-12, IL-1β, IL-4, IL-13, IL-6, IL-17, IL-1ra, IL-7, IL-9, eotaxin, IL-10, MIP1α, basic FGF, GCSF and VEGF compared to the CFS/ME patients and controls (p ≤ 0.04). There were no significant differences between groups for IL-8, MCP-1, MIP1β, RANTES, GMCSF, TNF-α, and IL-2. Conclusion: CFS/ME and MS patients both displayed abnormal cytokine levels, with dual expression of Th1 and Th2 cytokines. Further research into cytokines such as IFN-γ, IL-10 and IL-5, with the use of a specific CFS/ME case definition and sensitive cytokine assays, is required to improve the understanding of the pathophysiology of CFS/ME.

The Effects of Influenza Vaccination on Immune Function in Patients with Chronic Fatigue Syndrome/Myalgic Encephalomyelitis

Immune dysfunction is a hallmark of Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME). The purpose of this pilot study was to identify the effects of influenza vaccination on immune function in patients with CFS/ME. We included 7 patients meeting the Centre for Disease Control and Prevention criteria (CDC 1994) for ME/CFS and 8 control subjects. Bloods were collected from all participants prior to vaccination with Influvac, a trivalent inactivated influenza vaccine (TIV), 14 and 28 days following vaccination. The immune parameters examined include Natural Killer (NK) phenotypes, NK cytotoxic activity, FOXP3 and Th1/Th2/Th17 related cytokines. Flow cytometric protocols were employed. There was no significant difference in NK phenotypes and Tregs numbers between CFS/ME patients and healthy controls. However, NK activity was significantly decreased at baseline and at 28 days, while at 14 days it significantly increased in the CFS/ME patients compared to the healthy controls. Th1 pro-inflammatory cytokines increased considerably in the CFS/ME patients at 28 days compared to the non-fatigued controls. Only one Th2 cytokine, IL-4, increased in the CFS/ME participants. FOXP3 expressing Tregs only increased significantly at day 28 post vaccination in the CFS/ME patients compared to the healthy controls. Self-rated wellbeing was lower for patients at day 28 while at baseline and day 14 no differences were observed. In this pilot study immunization with influenza vaccine is accompanied by a degree of immune dysregulation in CFS/ME patients compared with controls. While vaccination may protect CFS/ME patients against influenza, it has the ability to increase cytotoxic activity and pro-inflammatory reactions post vaccination. The role of Tregs in promoting a toxic effect at 28 days post-vaccination in our patient group cannot be ruled out. The benefits of influenza vaccine still likely outweigh the risks CFS/ME patients experience following vaccination.

The Effect of Curcumin on Patients with Chronic Fatigue Syndrome/Myalgic Encephalomyelitis: An Open Label Study

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS), recently renamed as systemic exertion intolerance disease (SEID), is a chronic and often disabling disease. Although the exact pathophysiological mechanism of ME/CFS is unknown, immunological abnormalities may play an important role. Curcumin is a herb with powerful anti-oxidative, and anti-inflammatory properties. Therefore, we hypothesized that curcumin has favorable effects on symptomatology in ME/CFS patients. In total 52 patients participated, nine stopped the use of curcumin because of side effects. All remaining patients (n = 43) met the criteria for CFS;72% met the criteria for ME. Before and 8 weeks after the use of curcumin complexed with phosphatidyl choline, 500 mg bid, the CDC inventory for assessment of Chronic Fatigue Syndrome was filled in. The CDC questions (n = 19) were scored and divided into 2 parts: the first being specific for CFS complaints (n = 9), the second being scores of less specific symptoms (n = 10);denoted as CDC rest score. Results showed that 8 weeks curcumin use significantly decreased the CFS related symptom scores, but not the CDC rest scores. Analyzing the data separately for ME and CFS patients, the same significance for the CFS symptom scores was present. Conclusion: in this open-labeled study, 8 weeks curcumin use in a phosphatidyl choline complex reduced ME/CFS symptomatology. Therefore, a randomized placebo controlled study is warranted to assess its efficacy in ME/CFS patients.

Enhanced Gene Expression Following Vaccination in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis

Vaccines have been shown to cause differential expression of genes and increase antibody titers against antigens. Influenza vaccines may have an effect on unexplained disorders such as Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME). Immunological changes have been identified following immunization with trivalent influenza vaccine (TIV). The objective of this pilot study was to examine the consequences of TIV on cytokine and cytotoxic genes in CFS/ME. Peripheral blood mononuclear cells were preferentially isolated from whole blood of 7 CFS/ME patients and 8 controls. Following total RNA extraction and synthesis of cDNA, reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) was used to determine the expression levels of mRNAs for cytotoxic genes (perforin (PRF1), granzyme A (GZMA), granzyme B (GZMB) and cytokine genes. GZMB was significantly increased overall in the CFS/ME patients compared to the controls. GZMA was significantly increased 28 days after vaccination while PRF1 was reduced prevaccination but increased 14 days post-vaccination in the CFS/ME patients. There were no significant changes in cytokine genes pre or post vaccination. Administration of TIV may increase the expression of lytic genes in CFS/ME and this may contribute to the increase in cytotoxic activity we observed in these patients post vaccination.

Quantification of the Beneficial Effects of Compression Stockings on Symptoms of Exercise and Orthostatic Intolerance in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis Patients

Chronic fatigue syndrome and myalgic encephalomyelitis (CFS/ME) are, amongst others, characterized by exercise intolerance, pain, post exertional malaise and orthostatic intolerance. It has been shown in venous disease and sport participation that compression stockings may improve exercise performance and reduce post exercise muscle soreness. Moreover, its use is advocated in orthostatic hypotension. Therefore, it was hypothesized that compression stockings may reduce symptomatology in CFS/ME patients. Methods: 100 patients used compression stockings class II for minimally 3 weeks and thereafter filled in a questionnaire, based on the Rand 36 physical activity questions (n = 9), whether compression stockings changed perceived symptoms or not. Moreover, 7 questions referring to prolonged standing and sitting, to recovery post exercise, muscle pain during or immediately post exercise, and to dizziness/light-headedness during or immediately post exercise, while standing and during prolonged sitting were added. Questions were scored as 1: able to perform activity much less while wearing the stockings, 2: perform activity somewhat less, 3: no perceived change in activity, 4: perform activity slightly better, 5: able to perform activity much better while wearing the stockings. Results: In patients able to answer the question, all mean scores per activity were significantly higher than 3, being no perceived change in activity while wearing the stockings. Subgroup analysis showed that patients with orthostatic intolerance reported higher effects than patients without orthostatic intolerance. Conclusion: This pilot study suggests that compression stockings may be useful to reduce symptomatology of physical activities in CFS/ME patients, especially in patients with orthostatic intolerance. Larger prospective studies with hard endpoints are warranted.

Blood Volume Status in Patients with Chronic Fatigue Syndrome: Relation to Complaints

Four studies have compared a possible decrease in circulating blood volume in Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) patients when compared to a healthy population. A more recent study has proven a correlation between RBC volume and OI in chronic OI patients without being diagnosed ME/CFS. The aim of the present study was to relate measured blood, RBC and plasma volumes (absolute and percent normalized) with the orthostatic intolerance complaints in ME/CFS patients. In the included 11 female ME/CFS patients, percentage decrease in normalized blood, RBC and plasma volume was similar for all three components: 83% &plusmn;12%, 83% &plusmn;12% and 83% &plusmn;11%, respectively. In patients with a clinical suspicion of OI (n = 7) all 3 volume components were significantly lower compared to patients without clinical suspicion of OI (n = 4). The difference percentage to normalized Blood volume was 77(7) vs 94(10) (p-value < 0.02), difference percentage to normalized RBC volume was 76(7) vs 96(10) (p-value < 0.01) and difference percentage to normalized plasma volume was 77(7) vs 93(10) (p-value < 0.05) in OI present versus absent. Plasma volumes were plotted against RBC volumes: the relation found was RBC volume = 0.99* Plasma volume + 1.55;p < 0.001;r = 0.90. In line with literature data, this pilot study shows that total blood volume and its components: RBC and plasma volume may be reduced in ME/CFS patients, especially in the presence of a clinical suspicion of OI.

Decreased Expression of TRPM3 and mAChRM3 in the Small Intestine in Chronic Fatigue Syndrome/Myalgic Encephalomyelitis

Introduction: Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME) is often associated with gastrointestinal disturbance and inflammatory markers;however, there have been no histological studies performed in the small intestine from CFS/ME patients. The aim of this investigation was to assess the expression of certain inflammatory markers and inflammatory receptors, namely transient receptor potential melastin 3 (TRPM3) ion channels and muscarinic acetylcholine M3 (mAChRM3) receptors, in small intestinal tissues in a case controlled study comprising a CFS/ME patient and a healthy non-fatigued control. Method: Immunohistochemistry was performed on a small intestinal biopsy from a CFS/ME patient (age = 50;female) with self-reported symptoms of gastrointestinal disturbance and a non-fatigued control (NFC), (age = 28;female). Semi-quantitative analysis of expression was undertaken for interferon-gamma (IFNy), interleukin-1 alpha (IL-1α), tumour necrosis factor-alpha (TNFα), TRPM3 ion channels and mAChRM3 acetylcholine receptors. Results: There was significantly decreased expression of TRPM3 in the CFS/ME patient (35% &plusmn;9%) and a significant decrease in mAChRM3 in the CFS/ME patient (54% &plusmn;9%). There was no difference in IL-1α between CFS/ME patient and NFC, however;there was an increase in IFNy (13% &plusmn;6%) in the CFS/ME patient compared to NFC. There was a difference observed in TNFα in CFS/ME compared to NFC. Conclusion: Differences were noted in the expression of specific TRP ion channels and cholinergic receptors in CFS/ME compared with NFC, with CFS/ME demonstrating decreased TRPM3 and mAChRM3. Further, IFNy was increased, and TNFα decreased, in the small intestine of the CFS/ME patient with reported gastrointestinal disturbance.