The vasculitides are a group of rare diseases with different manifestations and outcomes. New therapeutic options have led to the need for long-term registries. The Rheumatic Diseases Portuguese Register, Reuma.pt, is a web-based electronic clinical record, created in 2008, which currently includes specific modules for 12 diseases and > 20,000 patients registered from 79 rheumatology centres. On October 2014, a dedicated module for vasculitis was created as part of the European Vasculitis Society collaborative network, enabling prospective collection and central storage of encrypted data from patients with this condition. All Portuguese rheumatology centres were invited to participate. Data regarding demographics, diagnosis, classification criteria, assessment tools, and treatment were collected. We aim to describe the structure of Reuma.pt/vasculitis and characterize the patients registered since its development.p = 1.00, p = 0.479). Biologic treatment was prescribed in 0.8% of patients with GCA, 26.7% of patients with AAV, and 7.6% of patients with BD. There were 34 (4.9%) deaths reported.A total of 687 patients, with 1945 visits, from 13 centres were registered; mean age was 53.4 ± 19.3 years at last visit and 68.7% were females. The most common diagnoses were Behçet’s disease (BD) (42.5%) and giant cell arteritis (GCA) (17.8%). Patients with BD met the International Study Group criteria and the International Criteria for BD in 85.3 and 97.2% of cases, respectively. Within the most common small- and medium-vessel vasculitides registered, median [interquartile range] Birmingham Vasculitis Activity Score (BVAS) at first visit was highest in patients with ANCA-associated vasculitis (AAV) (17.0 [12.0]); there were no differences in the proportion of patients with AAV or polyarteritis nodosa who relapsed (BVAS≥1) or had a major relapse (≥1 major BVAS item) during prospective assessment (Reuma.pt/vasculitis is a bespoke web-based registry adapted for routine care of patients with this form of rare and complex diseases, allowing an efficient data-repository at a national level with the potential to link with other international databases. It facilitates research, trials recruitment, service planning and benchmarking. The vasculitides are a group of relatively uncommon and complex diseases. The increased clinical trial activity and new therapeutic options for patients with vasculitis have led to the development of long-term specific registries for this condition , 2.www.reuma.pt) and currently includes specific modules for twelve different groups of rheumatic diseases  in Portuguese and English language. Over 20,000 patients and 170,000 visits have been registered, up to December 2018, by 79 national and international rheumatology centres. On October 2014, Reuma.pt launched a dedicated module to register patients with vasculitis. The variables included were chosen based on other pre-existing European registries, particularly UKIVAS (UK and Ireland Vasculitis Registry)  and C-reactive protein [CRP]), and imaging. A total of 63 ultrasounds were performed in 35 patients with an established diagnosis of GCA for monitoring purposes.p = 0.025) and cryoglobulinaemic vasculitis . There were no differences in cumulative damage at last visit between patients with AAV and PAN ; however, median [IQR] VDI of patients with cryoglobulinaemic vasculitis (1.0 [3.0]) was significantly lower in comparison to PAN (p = 0.001) and AAV (p < 0.001). In addition, no differences in disease activity at first visit and cumulative damage at last assessment were found amongst each subtype of AAV (p > 0.05). Prospective BVAS assessment was available for 12 patients with PAN (collected in 74 visits), 36 patients with AAV (collected in 125 visits) and 2 patients with CV (collected in 5 visits). In PAN, 6/12 (50%) patients had at least one disease relapse recorded (1/6 with a major relapse), in AAV, 18/36 (50%) patients had at least one relapse recorded (7/18 with a major relapse); and in CV, 1/2 (50%) patients had at least one relapse recorded (corresponding to a major relapse). No differences were found in the proportion of patients with a diagnosis of PAN or AAV who relapsed (p = 1.00) or had a major relapse (p = 0.479). Moreover, amongst each subtype of AAV, there were no differences found in the number of patients who relapsed  or who had a major disease relapse . In all vasculitides quality of life has been assessed by generic measures such as the SF-36, currently included in the AAV core set measures of assessment endorsed by the Outcome Measures in Rheumatology (OMERACT) Vasculitis Working Group  and GCA [n = 8]).Deaths were reported in 34 (4.9%) patients , at a mean age of 72.6 ± 15.5 [30–98] years and mean disease duration of 4.9 ± 5.7 [0–22] years; however, specific cause of death was available in only 10 cases, none of which directly related to treatment. Infection was the most common cause of death  , 37; Fran = 298), largely followed by GCA (n = 125). Although Reuma.pt/vasculitis is a registry reflecting the reality of the Portuguese Rheumatology centres and is generally not intended for epidemiologic studies, the number of patients with BD registered  exceeded the previously reported prevalence of 1.5 per 100,000 in 1991 [n = 94 vs. n = 72), reflecting modern practice and the incorporation of ultrasound as an essential diagnostic tool in the Rheumatology setting [Reuma.pt/vasculitis comprises a group of various subtypes of vasculitis, with different clinical manifestations, classification criteria and disease specific assessments and outcomes of interest. In addition, the registry of data by the physicians is done on an entirely voluntary basis. Therefore, balancing granularity and feasibility of data collection, when constructing the database, posed a challenge. Although we encountered missing data in our analysis, mainly on the prospective disease assessments and cause of death, overall our data was robust and allowed us to obtain reliable information on many of our patients’ characteristics. More than 1/3 of patients had the diagnosis of BD , very frequently encountered in this sort of registry, the diagnosis of each subtype of vasculitis was purely based on the submitting physician’s clinical knowledge and experience, therefore lacking external validation. In addition, most of the classification criteria used are currently outdated and waiting for the DCVAS results to ensure proper update . MoreoveWe expect this registry to continue expanding, including more centres, different specialties (e.g. Nephrology) and other data items. We aim to include the new AAV patient-reported outcomes (AAV-PRO) ; a speciWe anticipate that Reuma.pt/vasculitis will form the basis of many future national and international studies of vasculitis by allowing recruitment into trials, benchmarking and service planning. Its link to the biobank will facilitate translational research, and its connection to INFARMED will ensure proper report of adverse effects related to treatment. In addition, Reuma.pt/vasculitis will be an important source for longitudinal observational studies on the most relevant outcome measures of interest in vasculitis (e.g. treatment efficacy).Additional file 1: Figure S1. How to register the classification criteria of a patient with giant cell arteritis.


Tianma Gouteng Decoction (TGD) is widely used in traditional Chinese medicine for the treatment of hypertension and its related complications, but its mechanisms remain incompletely defined. We now aim to assess the protective effect of TGD against cardiovascular damage and to investigate its characteristics and underlying mechanisms. Blood pressure was determined in TGD-treated spontaneously hypertensive rats (SHR) by noninvasive measurements. Echocardiography was performed to assess cardiac function and structure and sirius red staining to evaluate cardiac fibrosis, and the degree of vascular remodeling was evaluated. Additionally, vasoconstriction and relaxation factor expression changes were examined by means of ELISA. Protein expression changes were verified by western blot. Compared with untreated SHR, TGD-treated SHR exhibited cardiovascular traits more akin to those of the normotensive Wistar Kyoto (WKY) rats. That is, they had lower diastolic blood pressure, systolic blood pressure and mean BP, and increased expression of vasodilation factor. We also found that TGD reduces ventricular and vascular remodeling and improves cardiac function in SHR. Finally, we tested the antiapoptosis effect TGD exerts in SHR, ostensibly by upregulating the expression of OPG, TRAIL, and death receptor 5 (DR5) and downregulating caspases 8, 7, and 3. TRAIL may also exert antiapoptotic and prosurvival effects by upregulating AKT expression. Therefore, TGD may reverse cardiovascular remodeling in SHR by upregulating the expression of OPG and TRAIL, upregulating AKT, and inhibiting apoptosis, at least in part. For the first time, we have shown that OPG and TRAIL play complimentary cardioprotective roles in SHR. Hypertension is a major public health issue worldwide, but the rates of treatment and control are still very low . In receOsteoprotegerin TNFSF11B, OPG) is a soluble member of the TNF superfamily and was first described as an important regulator of osteoclast formation 20 years ago . Recent B, OPG isUncaria, Gastrodia elata, Scutellaria baicalensis Georgi., Eucommia ulmoides Oliv., achyranthes root, Loranthus parasiticus, abalone shell, Gardenia, Leonurus japonicus, caulis polygoni multiflori, and Poria cocos. Chinese pharmacopoeia (2015 edition) reported that, in recent decades, TDG has been widely used in the clinical treatment of symptoms caused by hypertension, such as dizziness and headache [Gastrodia elata, can effectively intervene in RAAS and PPARγ, thereby exerting antihypertensive effect [Tianma Gouteng Decoction (TGD) is made up of 11 commonly used prescriptions  and Wistar Kyoto (WKY) (n = 5) rats (14 weeks old) were obtained from Vital River Laboratory Animal Technology Co., Ltd. . All the rats were kept in a temperature-controlled room (22°C ± 2°C) with 40% ± 5% humidity, exposed to ordinary lighting . The experimental procedures conformed to Directive 2010/63/EU of the European Parliament, and all animals were handled according to the guidelines of the TCM Animal Research Committee (TCM-LAEC2014005) of Tianjin University. SHR were randomly divided into three groups: (1) saline , (2) TGD 20 g/kg/d , (3) TGD 40 g/kg/d . A group of WKY rats  was used as a control.Male spontaneously hypertensive rats (SHR) , abalone shell 20 g (first fried), Gardenia 6 g, Scutellaria baicalensis Georgi 6 g, achyranthes root 12 g, Eucommia ulmoides Oliv. 12 g, Leonurus japonicus 12 g, Loranthus parasiticus 12 g, dried caulis of polygoni multiflori 12 g, and Poria cocos 12 g. All of the above ingredients were provided by the Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine. TGD is concentrated into granules; each 1 g granule is equal to 10 g of raw materials and complies with the guidelines of Good Manufacturing Practices and Good Laboratory Practices formulated by Chinese government agencies. The TGD was dissolved in pure water to prepare solutions having a concentration of 200 mg/mL or 400 mg/mL for the experiment.Tianma Gouteng Decoction from Saline was purchased from China Otsuka Pharmaceutical Co., Ltd. . The enzyme-linked immunosorbent assay (ELISA) kits, including prostacyclin (PGI2), thromboxane A2 (TXA2), angiotensin II (Ang II), and endothelin 1 (EDN1), were all obtained from Uscn Life Science, Inc. . The primary antibodies of GAPDH, AKT, p-AKT, caspase 8, caspase 7, caspase 3, and secondary antibodies were purchased from Proteintech Inc. . The antibodies of OPG, TRAIL, and DR5 were purchased from Abcam PLC .The blood pressure, heart rate, and bodyweight of the rats were monitored weekly on conscious animals by a noninvasive tail sleeve method using an animal sphygmomanometer . For each rat, blood pressure and heart rate were measured by multiple readings until 5 stable measurements were obtained continuously. The data were calculated as the average blood pressure and heart rate values. A conventional hundredth gram scale was used for weighing every week.The serum levels of PGI2, TXA2, EDN1, and Ang II were determined using the ELISA kits according to the manufacturer's protocol.All rats were examined by echocardiography at 12 weeks after initial drug administration. A Vevo 2100  ultra-high-resolution animal ultrasound imaging system was used to evaluate left ventricle function and structure, including systolic interventricular septum (IVSs), diastolic interventricular septum (IVSd), systolic left ventricular posterior wall (LVPWs), diastolic left ventricular posterior wall (LVPWd), ejection fraction (EF%), and left ventricular mass (LVmass).μm) were excised and stained with hematoxylin-eosin. The sections were analyzed under a light microscope. Images were captured by a digital camera connected to an optical microscope  and processed using the Leica Application Suite .After induction of anesthesia by intraperitoneal injection of 5% chloral hydrate (300 mg/kg), euthanasia was performed by excessive inhalation of isoflurane. The aorta was removed and fixed in a 4% paraformaldehyde solution for more than 48 hours, and then further prepared for paraffin sectioning. Serial sections (5 μm), and stained with sirius red. Images were captured by a digital camera connected to an optical microscope  and processed using the Leica Application Suite . To quantify the fibrosis, we used the image analysis system ImageJ  to calculate the proportion of red stained area within the frame.Similarly, the heart was dissected out and fixed in 4% paraformaldehyde solution for more than 48 hours, then further prepared for paraffin sectioning, cut into serial sections (5 n = 3). The whole left ventricle protein was extracted and subjected to radioimmunoprecipitation assay (RIPA) lysis buffer . The total protein concentration was determined using a bicinchoninic acid (BCA) protein assay kit (Tianjian Biotechnology (Beijing) Co., Ltd., Beijing, China) according to the instructions, and the average value of the concentrations was calculated. The sample was then stored at −80°C. Total protein was mixed with 3 μL of 5x SDS sample buffer. After separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (12% gel), the protein was transferred to a polyvinylidene fluoride membrane (PVDF membrane). After blocking with 5% skim milk powder or 5% bovine serum albumin (BSA) for 2 h, it was washed 4 times with Tris-HCl buffered saline (TBS) Tween for 7 min, and the membrane with the target protein was cut for incubation. The primary antibodies used were OPG, TRAIL, procaspase 8, cleaved caspase 8, caspase 7, procaspase 3, cleaved caspase 3, AKT, and p-AKT. All antibodies were diluted according to the manufacturer's instructions and left overnight at 4°C. Then, the membrane was washed 3 times for 5 min each and incubated with the secondary antibody for 2 h at room temperature and then washed 5 times with TBS Tween for 5 min each. Finally, the membrane was detected by ECL.The rats were sacrificed under anesthesia, and about 50 mg of myocardial tissue was taken out from the left ventricle and stored at −80°C  was considered to be statistically significant.Data presented are representatives from at least three independent experiments. Graph Pad Prism 5.0 and SPSS 23.0 were used to analyze all the data. All data were presented as mean ± SD. One-way ANOVA was used for multiple comparisons. A To evaluate the antihypertensive effect of TGD, we monitored the blood pressure of the animals for 12 weeks. The blood pressure of the SHR + saline group was significantly higher than that of the WKY group. After 12 weeks of administration of TGD, the TGD intervention groups  had reduced systolic blood pressure , diastolThe effects of TGD on cardiac function and structure were divulged by echocardiography. As shown by the M-mode echocardiograms , the lefTo further evaluate ventricular remodeling, we stained the rat heart with sirius red. Compared with the WKY + saline group, the left ventricle of the SHR + saline group had obvious collagen fiber deposition, suggesting a significant increase in cardiac interstitial fibrosis. After TGD treatment, cardiac interstitial fibrosis was significantly reduced in both SHR groups .The histological changes of the thoracic aorta were elucidated by Harris hematoxylin and eosin (H&E) staining. As shown in P < 0.01 (P < 0.01), which reduced the expression of vasoconstrictors TXA2 and Ang II  is widely used as a common prescription for headaches and dizziness in Chinese medicine. TGD has also been widely used in Chinese hospitals and clinics to treat symptoms related to hypertension. Compared with conventional antihypertensive drugs, TGD may be an effective alternative method for treating hypertension with fewer side effects . HoweverThe main finding of this study was that TGD lowered the blood pressure in SHR, especially after 8 weeks of administration. TGD at 40 mg/kg/d significantly reduced systolic, diastolic, and mean arterial pressure . FurtherTo study the cardiovascular protective mechanism of TGD, we evaluated the changes in cardiac function and structure, vascular remodeling, and changes in vasoconstriction and relaxation factors during the hypertension. Echocardiography demonstrates that TGD can reverse left ventricular remodeling and reduce heart weight in SHR but has no significant effect on EF. It can also improve cardiac fibrosis. In addition, HE staining showed that TGD treatment significantly reduced the thickening of the thoracic aorta in the SHR model, indicating that it reversed the effect of vascular remodeling. Therefore, we believe that TGD can exert cardiovascular protection by inhibiting heart and vascular remodeling, which are closely related to the reduction of blood pressure. Correspondingly, our results show that TGD treatment can significantly reduce the contractile factors TXA2, Ang II, and EDN1, which cause an increase in blood pressure in the SHR model and increase the expression of PGI2.κB ligand (RANKL) promotes VC, while osteoprotegerin (OPG) acts as a RANKL decoy receptor to block this effect, which is contrary to the known functional effects of these proteins in bone metabolism. However, in clinical studies, high levels of plasma OPG have been proven to positively predict the morbidity and mortality of CVD. Therefore, OPG plays a dual role in the cardiovascular system [The vascular calcification (VC) process can generally be described as progressive arteriosclerosis that results in reduced vascular elasticity leading to hypertension or atherosclerosis, ultimately increasing the risk of cardiovascular events . OPG is r system . In addir system . But TRAr system  and VSMCr system , but intr system , and stur system . This isr system , which mr system .In conclusion, our study indicates that TGD can increase the protein expression of OPG and TRAIL, activate p-AKT, and inhibit the caspase cascade, thereby inhibiting cardiomyocyte apoptosis and exerting cardiovascular protection. However, because TGD is composed of a variety of Chinese herbal medicines, it contains many compounds. The number of antihypertensive active ingredients in these compounds is unknown, and the signaling pathways through which they function are also unknown. Thus, in the course of developing better drugs for the treatment of hypertension, these important questions should be addressed through future research.


Mus musculus is useful for tracing prehistorical human movement related to the spread of farming. We determined whole mitochondrial DNA (mtDNA) sequences  of 98 wild-derived individuals of two subspecies, M. m. musculus (MUS) and M. m. castaneus (CAS). We revealed directional dispersals reaching as far as the Japanese Archipelago from their homelands. Our phylogenetic analysis indicated that the eastward movement of MUS was characterised by five step-wise regional extension events: (1) broad spatial expansion into eastern Europe and the western part of western China, (2) dispersal to the eastern part of western China, (3) dispersal to northern China, (4) dispersal to the Korean Peninsula and (5) colonisation and expansion in the Japanese Archipelago. These events were estimated to have occurred during the last 2000–18,000 years. The dispersal of CAS was characterised by three events: initial divergences (ca. 7000–9000 years ago) of haplogroups in northernmost China and the eastern coast of India, followed by two population expansion events that likely originated from the Yangtze River basin to broad areas of South and Southeast Asia, including Sri Lanka, Bangladesh and Indonesia (ca. 4000–6000 years ago) and to Yunnan, southern China and the Japanese Archipelago (ca. 2000–3500). This study provides a solid framework for the spatiotemporal movement of the human-associated organisms in Holocene Eastern Eurasia using whole mtDNA sequences, reliable evolutionary rates and accurate branching patterns. The information obtained here contributes to the analysis of a variety of animals and plants associated with prehistoric human migration.The Eurasian house mouse  Mus musculus) is a ubiquitous human commensal that is known to have spread through almost all of Eurasia with prehistoric human movements. The temporal and spatial aspects of the dispersal of mice, therefore, provide a practical way to interpret the process of human migration, agricultural development and cultural exchanges , M. m. domesticus (DOM) and M. m. musculus (MUS). These groups are believed to have originated in southern and western Asia, where they had a parapatric distribution in the central, western and northern portions of their home range, respectively , cytochrome b (Cytb) and NADH dehydrogenase subunit 3 (Nd3) genes, have been used in previous studies . We focused on the evolutionary history of the eastward movements of MUS-1 and CAS-1 in particular and aimed to precisely infer the divergence patterns and times of the major mtDNA lineages and elucidate the detailed spatiotemporal dynamics of MUS and CAS in northern and southern Eurasia.In this study, we determined the entire mitogenome sequences of 98 individuals collected across a broad geographic area covering 16 countries in Eurasia, which included four major mtDNA lineages of We used a total of 98 house mouse samples in this study. Most of our samples overlap with those used by Suzuki et al. . The samhttps://github.com/GregoryFaust/samblaster) with the ‘-M’ command option was used for identifying duplicates in read-id groups for exclusion from downstream analysis. The average median coverage of the whole genome sequence was 30.4 per sample. When reads were simultaneously mapped to the nuclear genome and mitogenome, the reads mapped to regions of the mitogenome that were highly similar or identical to regions in the nuclear genome, due to nuclear mtDNA segments, and yielded very low mapping quality (MQ) scores. To recalibrate the MQ score, we remapped all mapped mitogenome reads to the C57BL/6J complete mitogenome (NC_005089.1) using BWA-MEM and recalculated the MQ scores. Single-nucleotide polymorphisms and indels were obtained using the GATK4 HaplotypeCaller program . Our samples, along with their qualified concentrations and volumes, were sent to BGI  for whole-genome sequencing. Libraries were constructed for each sample with index sequences, and paired-end reads of 100 bp were sequenced using the BGISEQ-500 platform by BGI. For each sample, ~1 billion clean reads were obtained. We mapped the raw reads to the GRCm38 (mm10) house mouse reference genome sequence, including the mitogenome, using the BWA-MEM method . The principal advantage of this hypothesis-poor method over others that generate dichotomous branching networks or trees is that NN networks illustrate all potentially supported splits among a group of sequences as a reticulation. Further insights into the structure of each subgroup was obtained by constructing networks with mitogenome sequences  using the median-joining (MJ) method , MUS (n = 47) and CAS (n = 44) datasets, with substitution modes of GTR + G + I, HKY + G and HKY + G + I, respectively, which were determined using the Akaike information criterion, as implemented in MEGA. We performed 1000 bootstrap replicates to assess the robustness at each node. The time to the most recent common ancestor (tMRCA) of the mitogenome sequences  and 95% highest posterior density (HPD) was estimated using BEAST v1.8.4 software (Drummond and Rambaut http://beast.community/treeannotator) with the settings “Maximum clade credibility tree” and “Mean heights”, and were displayed using FigTree v1.4.3 software (http://tree.bio.ed.ac.uk/software/figtree/).A network of the complete mitogenome sequences was constructed using the neighbour-net (NN) method (Bryant and Moulton Cytb have been reported: 3.0 × 10−8 (lower rate) and 1.1 × 10−7 substitutions/site/year (higher rate) for ancient  and recent  divergences, respectively , suggesting an evolutionary rate of 2.4 × 10−8 substitutions/site/year for the mitogenome , as possible evolutionary rates. Furthermore, we considered alternative higher rates of 2.0 × 10−7 and 4.0 × 10−7 substitutions/site/year, as high rates are often used in studies on the population dynamics of M. musculus , haplotype diversity (Hd), number of polymorphic sites (S), nucleotide diversity (π) and mean number of pairwise differences among sequences (K) were estimated using DnaSP software (v5.00.7) , was estimated using the formula T = τ/2μk.The number of haplotypes  collected from Nepal, respectively . We obtained estimated divergence times of 520,000–580,000, 230,000–260,000 and 150,000 years ago for Nodes a, b and c, respectively  from northernmost China and the Russian Far East with an estimated divergence time of 8800  years ago  provides new insights into the evolution of M. musculus is thought to have occurred ca. 15,000 years ago  indicates a more recent expansion event with a predicted initiation time of 15,000 years ago  has presumably resulted from rapid expansion event that occurred in 130,000 years ago after the PGM. Meanwhile, inhabitation of western Europe by −7 evolutionary rate, we can propose an evolutionary history that agrees with archaeological events with known dates, as discussed below. On the other hand, the use of the highly accelerated evolutionary rate , as other haplotypes from Siberia (sample code: MG3004) and the Russian Far East (MG3026) contain almost identical sequences to those recovered from the westernmost sites in eastern Europe (MG3008).The present study provides information on the evolutionary scenarios driving dispersal events associated with prehistoric human movements, in particular those related to the eastward movement of MUS and CAS  is characterised by five episodic events (Steps I–V) of intermittent dispersal or population growth Fig. : (1) colM. musculus. For example, recent stowaway introductions of DOM mice (e.g. 50 generations ago) to the Japanese Archipelago are evident in haplotype structure analyses on the nuclear genome  from Ukraine was found to cluster with those from the eastern part of the western China  involved sudden population expansions in northern China, Korea and Japan, and agriculture-driven environmental change is considered a major factor shaping these sudden regional population expansions  and the other for the split in the Indian (Sri Lanka) haplogroup C1a2) with tMRCAs of ca. 8800 and 7500 years ago, respectively Fig. . The div with tMRsia Fig. .The expansion events predicted in Southeast Asia and East Asia coincide with the early stages of agricultural development in Asia (Diamond and Bellwood M. m. musculus type and M. m. castaneus type mtDNA in Japanese mice, they suggested a plausible connection to the model of human evolution in the Japanese Archipelago. Later, this human evolution model was called the ‘dual structure model’ by Hanihara (M. m. castaneus was assumed to arrive to the Japanese Archipelago with the Jomon people, and M. m. musculus was assumed to arrive to the Japanese Archipelago with Yayoi migrants. Kanzawa-Kiriyama et al. (When Yonekawa et al.  reportedHanihara . The majHanihara  and culta et al.  estimatea et al.  ‘dual stM. m. castaneus type mtDNA was relatively recent, yet during the late Jomon period. These two mouse migration periods are consistent with Saitou’s (In this study, based on mouse mitogenome data, we estimated the migration dates of mouse population movements of CAS-1 and MUS-1 to the Japanese Archipelago to be ca. 3500 and 2700 years ago, respectively see Fig. . The migSaitou’s  second aSaitou’s  proposedSaitou’s . WhetherM. musculus can be expected to provide valuable information on human colonisation and agricultural development, as shown in previous studies (e.g., Nunome et al. M. musculus based on whole mitogenome sequences. In particular, the divergence time estimates from our study are consistent with those reported in previous archaeological studies (Crawford and Lee −7 substitutions/site/year, as in this study, can provide relatively reliable estimates of divergence patterns and times for rodents in the Holocene.The early and middle Holocene is a crucial time period, when the fundamental regional human populations and cultures were established through colonisation and transfer of agricultural systems. In this context, the house mouse Supplementary figuresSupplementary Figure Table S1


Achieving the blood pressure treatment target in individuals with hypertension is a serious global health challenge. Furthermore, the actual burden of uncontrolled hypertension is poorly understood, especially in the developing countries. Therefore, this study comprehensively examined the prevalence and factors associated with uncontrolled hypertension in individuals receiving care at the primary healthcare facilities in the rural areas of Mkhondo Municipality in the Mpumalanga Province, South Africa.In this cross-sectional study, 329 individuals attending care for hypertension were recruited from January 2019 to June 2019 at three primary healthcare centres, namely, Piet Retief hospital, Mkhondo town clinic and Thandukukhanya community health centre. Uncontrolled hypertension was defined as systolic blood pressure ≥ 140 mmHg and/or diastolic blood pressure ≥ 90 mmHg in accordance with the South African Hypertension Society guideline (2014). Multiple logistic regression (Forward LR method) analysis was used to identify the significant determinants of uncontrolled hypertension.n = 187) with no significant difference between sexes, 57.38% male versus 56.88% female, respectively. In the multiple logistic regression model analysis after adjusting for confounding variables, obesity , physical activity  and HDL-C  were the significant and independent determinants of uncontrolled hypertension in the cohort.The majority of the participants were 55 years old and above (69.0%), Zulus (81.2%), non-smokers (84.19%) and had been diagnosed with hypertension for more than a year prior to the study (72.64%). The overall prevalence of uncontrolled hypertension was 56.83% (The high prevalence of uncontrolled hypertension in the study setting can be largely attributed to obesity, physical activity and dyslipidaemia. Treatment will require the collaborative efforts of individuals, clinicians and health authorities. All these determinants should be addressed decisively so as to achieve the treatment blood pressure targets in the study population. The prevalence of non-communicable diseases (NCDs) is increasing at an alarming rate world-wide and it has been driven by the rise in the incidence of cardiovascular risk factors such as obesity, diabetes and hypertension . HyperteHypertension is a multifactorial and highly complex disease that is characterized by a systolic blood pressure ≥ 140 mmHg and/ or diastolic blood pressure ≥ 90 mmHg . The risUncontrolled hypertension, defined as blood systolic blood pressure ≥ 140 and / or diastolic blood pressure ≥ 90 mmHg, has been associated with patients’ aging, clinical inertia, unemployed status and nutritional transitions . FurtherThis was a community based cross-sectional study conducted in the Piet Retief Hospital, Mkhondo town clinic and Thandukukhanya Community Health Center in Mkhondo Municipality of Mpumalanga Province, South Africa, from January 2019 to June 2019. These health care facilities provide chronic care services for the residents of Mkhondo Municipality. Mkhondo is located in the Gert Sibande district of the Mpumalanga province and is a resource-constrained community made up of a small town surrounded by farms and two townships (Ethandukukhanya and AJAX) with a population of approximately 189,036 .The study included individuals who were aged ≥18 years, had been diagnosed and initiated on treatment for hypertension at least a year prior to the study. Individuals who were bedridden, mentally compromised, pregnant or unable to give consent were excluded from the study.A sample size of 334 was estimated by using the formula for cross-sectional study:P = proportion with uncontrolled hypertension) of 68% was chosen for the sample size estimation. Z1 – α = 1.96 and D = absolute precision, which is taken as 0.05. Five participants dropped out of the study so a total of 329 patients were included in the final analysis.Given that the prevalence of uncontrolled hypertension ranged widely from 13.5–75.5% in South Africa , 14, P , secondary (7–12) and tertiary. Employment status was categorized as unemployed, employed or receiving social grants from the government. Smoking status were categorized as never smoked or ever smoked; while alcohol use was also categorized similarly. Physical activity was categorized as active . Also, participants reported their average consumption of fruits and vegetables, as well as fast food and salt intake.2) and was categorized based on WHO criteria as obese (30 or greater kg/ m2) or not  for each participant, was categorized according to the guidelines of The Society for Endocrinology, Metabolism and Diabetes of South Africa . All laboratory assays were conducted by the National Health Laboratory Services (NHLS) of Piet Retief and Ermelo Provincial hospitals in accordance with standard protocols.p-value of less than 0.05 was considered statistically significant.Data analysis was conducted using IBM SPSS Statistics for Windows, Version 25.0 . General characteristics of the participants were expressed as mean ± standard deviation for continuous variables. Categorical variables were reported as frequency (percentage). The associations between socio-demographic-, clinical factors and uncontrolled hypertension were examined by using bivariate analysis. Multiple logistic regression odds ratios and their 95% confidence intervals, using crude and adjusted logistic regression model analysis, helped identify the independent determinants of uncontrolled hypertension. A n = 329), 61 were males (18.5%) and 268 were females (81.5%). The majority of the participants were aged 55 years and above (69.0%), of Zulus origin (81.2%), non-smokers (84.19%), non-alcohol drinkers (77.81%), consuming fruit and vegetables (97.87%) and fast food about three times per week (61.09%), which differed according to sex. A sedentary lifestyle was reported by 231 participants (70.21%); 22 were males and 209 were females. Consumption of excessive amounts of salt was reported by 42 participants (12.78%) of whom a higher proportion were women (n = 37) in comparison with men (n = 5). The majority of the participants who had been diagnosed within the previous 5 years  were predominantly women (n = 194). Table Of the total number of participants (N = 329) had been on anti-hypertensive treatment for at least a year. The majority of the participants were on at least two or more anti-hypertensive drugs . Thiazide diuretic  was the preferred drug class, either alone  or in combination with other drugs  (86.62%). Successful treatment of reaching the blood pressure target of 140/90 mmHg occurred in 142 patients (43.62%) of the 329 participants. The overall prevalence of uncontrolled hypertension was 56.83% (n = 187), with no significant difference between sexes: 57.38% males versus 56.88% females, respectively , positive family history (p = 0.034), low HDL-C (p < 0.001) and obesity (p = 0.001) were significantly associated with uncontrolled hypertension. Other risk factors such as age, gender, smoking status, alcohol consumption, Total cholesterol, LDL-C and triglycerides were not significantly associated with uncontrolled hypertension (p > 0.05).In the bivariate logistic regression analysis Table , physicaIn the multiple logistic (crude and adjusted) regression model analysis Table , the catHypertension is an independent risk factor for cardiovascular diseases and all causes of premature deaths , 7, 18. South Africa has the highest prevalence of hypertension in Southern Africa . Despite2 [In the current study, the presence of obesity was significantly associated with uncontrolled hypertension. These findings are supported by studies conducted in Ethiopia , 27, Chi2 . The inc2 , 32. Pat2 , 34. A n2 .In the current study, engagement in physical activity was associated with the higher odds of having uncontrolled hypertension. It should be noted that the activity level of the participants was obtained by self-report and there was no standardized measure of the intensity of the physical activity. In addition, there is a lack of formal recreational centres (gymnasium) for graded exercises in the study setting. Also, other risk factors, such as excessive salt intake, may have masked the beneficial effect of exercise on blood pressure control in this population. A study conducted in Ethiopia showed that non-adherence to physical activity was associated with uncontrolled hypertension . In addiFurthermore, the current study found an association between decreased HDL-C (< 1 mmol/L) (dyslipidaemia) and uncontrolled hypertension. Dyslipidemia comprises having an abnormality in LDL-cholesterol, HDL-cholesterol and triglyceride levels . AbnormaAnti-hypertensive drugs are prescribed mainly to reduce blood pressure and complications associated with the disease. Studies have shown that following clinical guidelines in practice improves the treatment outcomes . AdditioThis study has provided very important insights into the treatment outcome (blood pressure) of individuals attending health facilities for hypertension in the region, but the limitations of the study cannot be ignored. Causal association of the determinants cannot be ascertained, given the cross-sectional design method adopted in the study. Also, the lifestyle assessment depended largely on self-report measures which can be subject to social desirability bias. In addition, the urinary sodium excretion level could have added more objectivity about the consumption of salts among the study participants.We found a high prevalence of uncontrolled hypertension in the study setting, possibly attributed to obesity, physical activity and dyslipidaemia. All these determinants should be addressed decisively through collaborative efforts of individuals, clinicians and health authorities. Such collaboration would facilitate achieving the treatment blood pressure targets in the study population. To the best of our knowledge, this study is the first to report the prevalence of uncontrolled hypertension with an association between it and low HDL-C, inadequate physical activity plus obesity among residents of Mkhondo municipality. Furthermore, this study has highlighted the need to implement more aggressive health strategies targeting the identified determinants as well as blood pressure of the patients in order to improve the overall outcomes of care in the rural communities of Mkhondo municipality of Mpumalanga province.Additional file 1. Mpumalanga Data Collection Sheet


Along with a nutritional transition in Sub-Saharan Africa, the prevalence of non-communicable diseases is increasing rapidly. We assess the association between food intake and cardiometabolic risk factors in a rural population in Uganda.The present study was based on data from a household-based case-control study of diabetic and non-diabetic households in Southwestern Uganda, 2012–2013. We analysed food intake in 359 individuals age ≥ 13 years from 87 households, using a household food frequency questionnaire, and measures of glycated haemoglobin (HbA1c), height and weight. We used multinomial logistic regression to model abnormal HbA1c (≥5.7%) and weight status  as an outcome of total food intake and by nine food groups. Results were reported as odds ratios (OR) with 95% confidence intervals (CI). Models were adjusted for three nested sets of covariates.The diet primarily consisted of staple food (cassava and plantain). High-Glycaemic Index staple food was the most consumed food group . Milk, meat, fish and vegetables were the least consumed food groups (medians: 0–3 servings/week). Median intake of sugary food was 6 servings/week (p25-p75: 2–9). The OR of having abnormal HbA1c or being overweight increased with every weekly serving of food . Of specific food groups, each weekly serving of meat increased the OR of being overweight with 33% (95% CI: 1.08–1.64), and fruit intake decreased the OR of abnormal HbA1c , though this latter association was attenuated after adjustment for weight status, aerobic capacity, and socioeconomic status.Diet was monotonous, mainly consisting of cassava and plantain, and increasing food intake was associated with abnormal HbA1c and overweight. To prevent non-communicable diseases a diet with higher intake of fish and vegetables, and less sugary food is recommended.The online version contains supplementary material available at 10.1186/s13690-021-00547-x. Low- and middle income countries (LMIC) are experiencing a nutritional transition characterized by traditional diets being replaced by energy-dense and processed food and beverages . In Sub-Historically, the diet in SSA consisted of fruits, green leafy vegetables, legumes and whole grain ; food itIn Uganda, the diet typically includes plantain, maize, cassava, millet, yams, peanuts, pumpkin, beans, green vegetables, and different fruits, supplemented by smaller amounts of meat and fish . HoweverThe effect of food intake on cardiometabolic risk factors has been heavily investigated in high income countries (HIC). However, the increasing prevalence of cardiometabolic conditions in SSA , 10 wherThe aim of the present study was to investigate food intake and its association with cardiometabolic risk factors in a rural Ugandan population potentially being in the midst of a nutritional transition.n = 45), self-reported malaria (n = 4), sickle cell disease (n = 1) and those being pregnant (n = 12) as these conditions (or treatment) may influence the association between food intake and glycated haemoglobin (HbA1c) and weight status. Furthermore, we excluded 12 participants due to missing information about food consumption and 4 participants due to missing information about aerobic capacity. This resulted in an analytical sample of 359 participants.We conducted cross-sectional observational analyses using data from a case-control household study in Kasese District, Southwestern Uganda in 2012–2013, with cases being households including a member diagnosed with T2D and controls being households without any members with diagnosed T2D [2-max) [HbA1c (%), height (cm) and weight (kg) and estimated aerobic capacity (VO2-max)  was meas2-max) .We categorised food intake according to the Food and Agriculture Organization of the United Nations’ (FAO) food groups , but in We used HbA1c as a proxy for diabetes risk. The American Diabetes Association recommends cut-off level for diagnosing diabetes at HbA1c ≥ 6.5%, while HbA1c ≥ 5.7 is considered prediabetes . In the 2 was used to define weight status for adults while we used WHO’s sex and age specific BMI z-score (BMIZ) for participants aged ≤19 years [n = 8), overweight and obese were merged into one group.Body mass index (BMI), calculated as weight(kg)/height(m)19 years . Weight 19 years , 19. DueWe included aerobic capacity and household SES as co-variates. Aerobic capacity was categorised into three groups  according to Astrand . HousehoCharacteristics of the study participants were reported as proportions or means with 95% confidence intervals (CI) for the total study population and by sex. Food intake was visualized for the total study population as boxplots including median, p25, p75, minimum and maximum values for the nine food groups. To model HbA1c or weight status as a function of total food intake and separate food groups, we used multinomial logistic regression with household as a random effect to account for household clustering. All models were adjusted for sex and age (Model I). As a next step, we adjusted for total food intake (Model II) and lastly we ran a model (Model III) also adjusting for aerobic capacity. In the analysis of HbA1c, weight status was included as a co-variate in Model III. The results were presented as odds ratios (OR) with 95% CI. The analyses were conducted using STATA .Overall, the study population included 57.9% women, mean age was 35.5 years (95% CI: 33.4–37.6) and 41.5% of the participants shared household with a person with T2D . High-GI staple food was the food group with the highest median of weekly servings   with each food serving per week. Adjusting for weight status, aerobic capacity and socioeconomic status did not attenuate the association  increased OR of being overweight compared to normal weight per one additional serving of food per week , though differences in portion sizes and number of servings between household members may have caused imprecise reporting of food intake. Secondly, the FFQ as well as the food groups used in the present study was designed to assess dietary diversity, not cardiometabolic risk factors . ThirdlyWe used data from a case-control household study. Even though participants with diagnosed T2D were excluded from the present study, participants having a household member diagnosed with T2D may have had higher predisposed genetic risk of abnormal HbA1c and may have changed their diet over time according to dietary recommendations for T2D. However, additional sensitivity analysis did not show any different results, when excluding households with T2D (results not shown). Also, in a previous study in the same study population , we founData were collected in 2012–2013, however, we are not aware of major political or economic changes or reforms that would have influenced agriculture, crops or eating patterns.In conclusion, we found a monotonous diet in our study population including protective components for cardiometabolic diseases, such as low consumption of meat and relatively high consumption of legumes and fruits. Despite a low meat intake, a 33% increased OR of overweight was found with each additional weekly serving of meat. Fruit intake was associated with decreased OR of abnormal HbA1c, though the association was attenuated after adjustment for weight status, aerobic capacity and socioeconomic status. Several risk factors for cardiometabolic disease was also observed in the diet, such as a relatively high consumption of sugary food, and low consumption of vegetables. Thus, the rural Ugandan population is most likely gradually transitioning towards a more unhealthy diet. In Uganda, nutrition strategies in general and those to prevent cardiometabolic diseases should focus on how people in rural areas can access and utilize existing resources sustainable e.g. fish from local lakes and grow crops that will provide both quantity, quality and a varied diet.Additional file 1.Additional file 2: Table 1. Food items included in food groups in Kasese District, Uganda (N=359), Food Intake and Cardiometabolic Risk Factors in Rural Uganda study, 2012–2013


No effects were found for timed up and go test performance, muscle strength, and frailty status. The community program exhibited good reach, effectiveness, adoption, and implementation. Our study demonstrated that FPT was associated with greater improvement in physical function in pre-frail/frail participants as compared to exercise activities offered at local senior activity centers. It is a feasible intervention that can be successfully implemented for frail older adults in their neighborhoods. Translation of community-based functional training for older adults to reduce frailty is still lacking. We evaluated the effectiveness and implementation of a community-delivered group-based functional power training (FPT) program for frail older adults within their neighborhoods. A two-arm, multicenter assessor-blind stratified randomized controlled trial was conducted at four local senior activity centers in Singapore. Sixty-one community-dwelling older adults with low handgrip strength were randomized to intervention (IG) or control (CG) group. The IG underwent the FPT program  delivered by a community service provider. The 12-week program comprised 2 × 60 min sessions/week. CG continued usual activities at the centers. Functional performance, muscle strength, and frailty status were assessed at baseline and 3 months. Program implementation was evaluated using RE-AIM framework. The program was halted due to Coronavirus Disease 2019-related suspension of senior center activities. Results are reported from four centers, which completed the program. IG showed significantly greater improvement in the Short Physical Performance Battery test as compared to CG (Trial registration ClinicalTrials.gov, NCT04438876. Registered 19 June 2020–retrospectively registered.The online version contains supplementary material available at 10.1007/s11121-021-01221-y. Community-dwelling older adults are prone to developing frailty  at high velocity (as far as possible) with a slow eccentric phase (force production by lengthening of muscles) to achieve the greatest benefit of muscular power and strength. Such training is effective and has emerged as an alternative modality of resistance training to preserve function of older persons to perform activities of daily living that often require quick, forceful motions  program using bodyweight, resistance bands, and chairs for frail older adults at a local senior activity center . Seven senior activity centers located within the residential estates in Singapore were approached, and the study was conducted at four centers. To be eligible for the study, participants had to (a) be aged 55 years and older, (b) have low muscle strength, (c) ambulate without human assistance and have no other physical limitations to participation and adherence to exercise, and (d) be able to understand basic instructions. Low muscle strength was defined as handgrip strength (HGS) less than 26 kg and 18 kg in men and women, respectively, according to the Asian Working Group for Sarcopenia (AWGS) 2014 consensus . Hourly sessions were held twice weekly at local senior activity centers. The exercise intervention comprised progressive power and balance exercises that targeted both upper and lower body muscles. The list of specific exercises for each respective session is presented in Table https://empower.org.sg/frailty-solutions/). Participants performed three sets of each power exercise, with 10–20 repetitions per set and 12–60 repetitions per set for balance exercises. For the power training, body weight and/or resistance bands were used as resistance and participants were instructed to move as fast as they can during the concentric phase and slowly during the eccentric phase (approximately 3 s) of the exercise movements. To ensure safety, blood pressure of participants was measured before and after each session with an automated sphygmomanometer . Arterial blood oxygen saturation and heart rate were measured at start, mid-point, and end of each exercise session with pulse oximetry  to monitor exercise intensity levels. Participants with either (a) blood oxygen levels below 95% saturation; (b) high resting heart rate (≥ 90 beats per minute); (c) abnormal resting blood pressure (systolic blood pressure ≥ 130 mmHg or diastolic blood pressure ≥ 80 mmHg); (d) giddiness or (e) any form of discomfort did not initiate or continue with training session. Participants also rated their perceived exertion after each exercise set and were encouraged to alert the trainer if they felt any discomfort during the training. We monitored and recorded adverse events throughout the intervention program. In addition, participants also received monthly health education talks on nutrition and cognition, conducted by staff from Geriatric Education and Research Institute.The IG underwent a 12-week structured FPT program, conducted by an exercise-science qualified trainer from a community service provider " in the last 6 months. Slowness was determined by 6 m walking speed with specified cut-offs based on gender and height. Exhaustion was self-reported through a 3-item questionnaire adapted from the SF-12 survey , and waist circumference were taken during the baseline assessment session. The Mini-Mental State Examination was administered to assess cognitive function in participants. It was scored out of 30, with higher scores indicating better cognitive function  using a power of 0.90 and error probability of 0.05, a sample size of 50 participants is required for each group to detect an assumed 20% difference in SPPB between IG and CG. In addition, with an assumption of 15% dropout rate, a sample size of 120 participants was initially targeted.t tests for continuous variables and chi-squared tests for categorical variables.Differences in baseline measures between CG and IG were compared using independent sample Levene’s test was conducted to ensure no violation of equal variance assumption. Linear mixed-effect modeling was performed to examine changes in physical function and frailty status between baseline and 3-month follow-up across both groups. Primary independent variables in each model included treatment groups, time, as well as group × time interaction. The models included random intercepts to account for correlations between repeated measures for each participant and were adjusted for age, gender, and physical activity levels. Clustering effect at the center level was omitted as intercepts did not vary significantly across centers. Data analyses were performed based on the intention-to-treat principle  or IG (n = 30). In partnership with a fifth senior activity center, another 50 individuals were screened, 33 completed pre-participation medical examination, and 31 eligible signed informed consent on 4 Feb 2020. The first case of Coronavirus Disease 2019 (COVID-19) in Singapore was confirmed on 23 January 2020. We could not proceed with the study after health authority mandated suspension of all group senior activities with effect from 8 Feb 2020 and the decision was made in May 2020 (with suspension still in force) to close the study and perform analyses on the 61 participants from the four centers. Post hoc power analysis revealed that this sample size was adequate to detect effects of moderate size. Four participants in the CG and seven participants in the IG did not complete the study. Table Figure F = 4.320, p = 0.043]. However, no significant interaction between group and time was found for frailty status F = 0.225, p = 0.637], HGS , KES , and TUG . Among the components of SPPB, group × time interaction effect was found only for repeated chair stands .The outcome measures at baseline and 3-month follow-up are presented in Table p = 0.010. Similarly, SPPB scores improved by 0.52 points (4.8%), 0.58 ES, 95CI , p = 0.008. Among the SPPB components, only repeated chair stand component was found to significantly improve by 0.42 points, 0.85 ES, 95CI , p = 0.002. Improvement of 0.58 s in TUG performance did not achieve statistical significance, 0.25 ES, 95CI , p = 0.071. No significant differences were found for HGS and KES. For the CG, significant improvement between baseline and 3-month follow-up was found in frailty status by 0.28 points, 0.40 ES, 95CI , p = 0.034; and HGS by 1.19 kg, 0.28 ES, 95CI , p = 0.020. No significant changes were found in KES, TUG, and SPPB performance for the CG.Post hoc pairwise comparisons found significant changes in outcome measures between baseline and 3-month follow-up in both groups. For the IG, post-program frailty status significantly improved by 0.36 points, 0.51 ES, 95% confidence interval (95CI) , A total of 160 older adults were assessed for eligibility, of which 34 (21.3%) did not meet the study’s inclusion criteria. Out of the remaining 126 (78.7%), 33 (26.2%) were deemed not suitable to participate in the intervention by a medical doctor. Among the 93 eligible individuals, one (1.1%) declined to participate. Therefore, 98.9% of eligible individuals agreed to participate in the FPT training program. However, 31 participants could not start the intervention due to the study being disrupted by COVID-19.The effectiveness of the intervention on objective physical outcome measures was reported above. Among the 27 participants who started the intervention, four 14.8%) did not complete the exercise program. One withdrew from the study due to personal reasons and one dropped out because of medical reasons unrelated to the study. There was no other adverse event during the program. Two participants reported that the exercise intervention was too challenging and dropped out on the fourth and fifteenth session, with attendance rate of 25% and 57.1%, respectively. The participant experience questionnaire was administered to 23 participants who completed the intervention. All participants agreed that the intervention was positive in terms of organization, engagement, and relevance to daily activities  of the participants completed all program sessions. On average, participants were found to complete 95.7% of the prescribed exercise program in the sessions they attended. Reasons for failing to complete the program include being late, feeling unwell, and leaving the session early. Seven (30.4%) participants achieved 100% participant fidelity rate.At the provider level, among the total of 96 sessions conducted across four senior activity centers, two (2.1%) sessions had minor adaptations by the trainer. One session reduced the number of repetitions for one set of power component exercise and the other session prescribed different balance exercises of lower difficulty. At the participant level, the mean attendance rate attained was 87.5% among those who completed the program. Most (87%) participants achieved an attendance rate of at least 80%, and 17.4%  are more closely related to muscle power than strength in older adults . A strategy to bridge the digital divide of some older adults has been to pair them up with others in their family or neighbors who can help with digital access and engagement with the live online training program. While not specifically measured in our study, the social engagement has been shown to be an important aspect of adherence to group training Below is the link to the electronic supplementary material.


Salmonella spp. in raw and ready-to-eat (RTE) foods, and Campylobacter spp. in the retail raw chicken meat collected in two counties of Transylvania, Romania. A total of 13.1% (51/388) of the examined food samples were found to be Salmonella positive, with a distribution of 14.7% (48/326) in the raw food  and 4.8% (3/62) in the RTE samples , respectively. These differences were statistically significant (p = 0.034). The isolates were serotyped as Salmonella Infantis (n = 19), Salmonella Typhimurium (n = 11) Salmonella Rissen (n = 8), Salmonella Derby (n = 3), Salmonella Enteritidis (n = 3), Salmonella Bredeney (n = 2), Salmonella Brandenburg (n = 1), Salmonella Gloucester (n = 1), Salmonella Goldcoast (n = 1), Salmonella Kottbus (n = 1), and Salmonella Ruzizi (n = 1). Campylobacter strains were present in 29.4% (10/34) of the investigated chicken samples, and the identified species were Campylobacter coli (70%) and C. jejuni (30%). From the 14 tested antimicrobials, the Salmonella isolates were resistant against azithromycin (88.2%), tetracycline (54.9%), sulfamethoxazole (54.9%), ciprofloxacin (45.1%), nalidixic acid (43.1%), ampicillin (35.3%), chloramphenicol (33.3%), tigecycline (25.5%), cefotaxime (13.7%), colistin (13.7%), trimethoprim (7.8%), and gentamicin (2%), resulting in the expression of 21 multidrug-resistant (MDR) profiles. Of 10 Campylobacter isolates, 80% were resistant to ciprofloxacin and nalidixic acid, 40% to tetracycline, and 10% to streptomycin and erythromycin, respectively. Our findings indicate that Romanian isolates of Salmonella spp. and Campylobacter spp., contaminating animal-origin foods, can exhibit MDR patterns, representing a public health risk.The survey was undertaken to investigate the presence and antimicrobial susceptibility profile of  Salmonella spp. and Campylobacter spp. are recognized as two of the most important foodborne pathogens that can cause severe infections in humans and economic losses worldwide. Their presence is monitored in different steps of the food chain, and especially in finished raw and ready-to-eat (RTE) products, as a safety criterion for the consumer, representing a very important tool for implementing efficient food safety systems  have accelerated the emergence and spread of antibiotic-resistant foodborne bacteria. Nowadays, the antimicrobial resistance (AMR) phenomenon is considered one of the most worrisome public health concerns, with negative impact on the effectiveness of public health interventions (Zhang The European Union (EU) member states make great efforts to establish harmonized interinstitutional strategies under a One Health approach to combat AMR. The collection of comparable, up-to-date, and reliable data is a prerequisite for the implementation of effective risk management measures by assessors. In this regard, baseline information about the occurrence of AMR zoonotic bacteria from the human–animal–food interface in Europe is published yearly by the European Food Safety Authority (EFSA) and European Center for Disease Prevention and Control, within a summary report  in different food products, from two Transylvanian counties of Romania.This study aimed to provide data on the occurrence and antimicrobial susceptibility profile of two major foodborne pathogens (n = 146), chicken carcasses (n = 98), and shell eggs  provided from private production units were screened for the presence of Salmonella spp. within the self-control process of each batch, in conformity with Regulation (EC) 1441/2007 (EC 1441/2007). Similarly, 62 RTE products, including sausages (n = 37), ham (n = 9), and salami (n = 16), and fresh chicken meat samples (n = 34), randomly collected from retail markets by veterinary inspectors within the official organized controls, were analyzed, to detect Salmonella spp. In addition, the retail chicken samples were monitored for the presence of Campylobacter spp.The study was undertaken between 2016 and 2018 in two counties of the historical Transylvania region, located in central Romania. A total of 292 raw food samples of pork  and Rambach (Biokar Diagnostics) mediums were used and the inoculated plates were incubated afterward at 37°C for 24 h. The isolation of Campylobacter spp. from the enriched samples was carried out on Columbia agar supplemented with sheep blood (Oxoid) at 42°C for 48 h under microaerobic conditions. For each positive sample, a single randomly selected bacterial strain was submitted to the Institute for Hygiene and Public Health  for antimicrobial susceptibility testing and serotyping in case of Salmonella isolates. Serotyping was carried out in accordance with the Kauffmann–White scheme by microtiter slide agglutination using polyvalent O- and H-antisera .® microbroth dilution system , in accordance with the manufacturer's instructions, and following the International Organization for Standardization  and pork [19.7%  Salmonella contamination level registered in pork, compared with chicken in this study is in contrast with the results published by several authors  is a common finding, which has been previously pointed out by many authors  Salmonella detection rate in these foods compared with raw foods. Nevertheless, the findings highlight a possible undercooking, contamination from raw materials, or food handlers of RTE products  were found to be Salmonella Typhimurium [reported as the most frequently isolated serotype from humans in Romania , according to which all these serotypes are typical of the tested products. Other rare serotypes, such as Salmonella Gloucester, Salmonella Goldcoast, and Salmonella Kottbus were recorded for the first time in Romania, emphasizing the spreading of sporadic serovars in our country.The recorded dominance of Salmonella strains, 92.2% were resistant to at least one antibiotic. Moreover, 68.6% (35/51) exhibited multidrug resistance. Only four pork origin strains were susceptible to all tested drugs , TET (54.9%), SMX (54.9%), CIP (45.1), and NAL (43.1%). These resistance rates  are consistent with previous reports for both chicken  in different combination forms (n = 21) highlight an urgent need for the implementation of efficient antimicrobial stewardship programs in animal husbandry. On the contrary, the lack of AMR toward MEM and CAF can constitute a promising tool for clinicians in the management of human salmonella infections.Similar to our results, increasing AMR trends were published for TET in China [87.5%  in this survey highlight that raw chicken meat can greatly contribute to the human campylobacteriosis cases, the most commonly reported foodborne infections in the EU since 2005.The detection rate of Campylobacter strains in Romania [31.8% for TET, 9.1% for CIP and NAL, 0% for STR , NAL (80%), and TET (40%) were found. Only one strain (10%) was resistant to STR and ERY, and all strains were susceptible to GEN. These resistance levels are higher than those that had been previously recorded for chicken origin Salmonella and Campylobacter strains, constituting a potential public health risk. Likewise, the findings could reflect an urgent need for the implementation of efficient antimicrobial stewardship programs in animal husbandry, and indirectly can constitute a promising tool for clinicians in the management of human salmonella and campylobacter infections. Further investigations are recommended to a better understanding of the complex puzzle of AMR phenomenon of the foodborne pathogens in our country.The results of this study showed that the investigated animal-derived foods from Transylvania region, Romania, can harbor MDR


Aedes larval indices. Therefore, the current study aimed at developing threshold values for vector indices in two dengue high-risk districts in Sri Lanka.  Early detection of dengue epidemics is a vital aspect in control programmes. Predictions based on larval indices of disease vectors are widely used in dengue control, with defined threshold values. However, there is no set threshold in Sri Lanka at the national or regional levels for Aedes aegypti [BIagp], and Ae. albopictus [BIalb]), of ten selected dengue high-risk Medical Officer of Health (MOH) areas located in Colombo and Kandy districts, were collected from January 2010 to June 2019, along with monthly reported dengue cases. Receiver Operating Characteristic (ROC) curve analysis in SPSS (version 23) was used to assess the discriminative power of the larval indices in identifying dengue epidemics and to develop thresholds for the dengue epidemic management.  Monthly vector indices , Moderate Risk (3.8 ≤ BIagp < 5), High Risk (BIagp ≥ 5), along with BIagp 2.9 ≤ BIagp < 4.2 (Low Risk), 4.2 ≤ BIagp < 5.3 (Moderate Risk), and BIagp ≥ 5.3 (High Risk) for Kandy. Further, 5.5 ≤ HI < 8.9, 8.9 ≤ HI < 11.9, and HI ≥ 11.9 were defined as Low Risk, Moderate Risk, and High Risk average thresholds for HI in Colombo, while 6.9 ≤ HI < 9.1 (Low Risk), 8.9 ≥ HI < 11.8 (Moderate Risk), and HI ≥ 11.8 (High Risk) were defined for Kandy.  Only HI and BIAe. aegypti and HI could be recommended as indicators for early detection of dengue epidemics and to drive vector management activities, with the objective of managing dengue epidemics with optimal usage of financial, technical, and human resources in Sri Lanka. The defined threshold values for  Aedes aegypti and Ae. albopictus are considered as efficient vectors for transmission of arboviral diseases such as dengue, chikungunya, and zika  (number of positive containers with Aedes larvae per 100 houses) and Container Index [CI] (number of positive containers with Aedes larvae per 100 containers) were calculated in accordance with the WHO guidelines [P < 0.05 at 95%) were considered as the discriminative indices to define dengue epidemics.House Index [HI]  to identifying dengue epidemics. The area under the curve (AUC) measures the total two-dimensional area that lies under the ROC curve. It was used to classify larval indices into three groups: (i) Noninformative (AUC ≤ 0.5), (ii) Moderately accurate (0.5 < AUC ≤ 0.7), and (iii) Highly accurate (0.7 < AUC ≤ 1), by modifying the recommendations of Luo et al. [Receiver Operating Characteristic (ROC) curves were used to assess the discriminative power of three larval indices  or in other words the points on the ROC curve located farthest from the line of equality was selected as threshold values based on the Youden index, to discriminate different phases in the dengue incidence . In thisEthical approval was obtained from the Ethics Review Committee of the Faculty of Medicine, University of Kelaniya (P/155/10/2015).Ae. aegypti (BIagp) and Ae. albopictus , HI, and CI from January 2010 to June 2019 are illustrated in S1 appendix. Seasonal fluctuation pattern of larval indices was observed in the districts of Colombo and Kandy  had the significantly strongest association with dengue cases reported in all MOH areas after lag periods of one (0.387 ≥ r ≤ 0.580) and two (0.587 ≥ r ≤ 0.767) months (P < 0.05). On the other hand, HI had a significantly negative association with dengue cases after one- and two-month lag periods , except in Akurana (r = 0.385). In summary, only BIagp and HI denoted significant associations with the incidence of dengue cases, and therefore, the above two indices could be used as important parameters for epidemic management via vector control approaches.According to the Pearson correlation analysis,  periods . The higagp of Akurana (0.530) and Moratuwa (0.507) had moderately accurate discriminative powers in reflecting dengue epidemics as shown in agp of Moratuwa (0.794), Akurana (0.761), and Kaduwela (0.731) denoted highly accurate discrimination of dengue epidemics, while moderate discriminative power was shown by BIagp in all the other study areas , respectively. In the case of HI, the minimum “Low Risk” cutoff value (4.7) was suggested for Piliyandala MOH area (located in the district of Colombo), while the minimum “Moderate Risk” (7.7) and “High Risk” thresholds were identified for Akurana MOH in Kandy. The Piliyandala MOH area had the maximum “High Risk” threshold of 16.3 for HI, while Gampola had the maximum “Low Risk (7.8)” and “Moderate Risk (12.1)” thresholds  directly influences the disease incidence and nature of dengue epidemics [A diverse interplay of multiple factors such as humans, pidemics , 40–42. pidemics . Considepidemics , 20, 24.Ae. albopictus was relatively higher in all study areas, a nonsignificant association was indicated by BIalb during both correlation and ROC analysis. Recent studies conducted in Sri Lanka have also evidenced high prevalence rates of Ae. albopictus reflected by the BIalb, which has been concluded as the vector that maintains the dengue viral strains, but with a lesser contribution towards transmission to humans [Ae. albopictus was abundant even in the urban and semiurban areas such as Kolonnawa, Kandy Municipal Council (KMC), Piliyandala, and Gampola. The presence of adequate vegetation and potential breeding sites could be the leading factors for the occurrence of Ae. albopictus [Ae. aegypti indicated a positive moderate to strong association with the reported dengue cases in all MOH areas after a lag period of two months. Therefore, Ae. aegypti could be recognized as the major transmitting agent of dengue within the studied MOH areas, as depicted by the higher association of BIagp with the epidemics of dengue [agp could be recommended as the key larval index that reflects the incidence of dengue epidemics.Even though the density of o humans , 43, 44.bopictus , 43. On Ae. albopictus and Ae. aegypti. A notable negative association has been observed among the vector densities of Ae. albopictus and Ae. aegypti in the study areas. This is evidenced by the decrease of BIalb, while BIagp increased prior to and within dengue epidemics [Ae. albopictus declines. This may be the reason for the negative association among the HI and dengue outbreak incidence within the studied MOH areas. The efficacy of CI is questionable due to some limitations such as incompetency to reflect the number of positive containers per house, per area, per person, and qualitative nature, implying a nondiscriminative power in defining dengue epidemics [agp and HI were recognized as reflective indices to predict dengue epidemics in the investigated study areas. The findings of the present study are also in agreement with previous investigations conducted in Sri Lanka [Irrespective of using both CI and HI in routine entomological surveillance, only HI stated a significant and moderate relationship with dengue cases. In general, the HI is influenced by densities of both pidemics , which ipidemics . Therefori Lanka , 44, 45.agp and HI individually at the regional level (MOH level). For Colombo district, 2.4 ≤ BIagp ≤ 3.7, 3.8 ≤ BIagp < 5, and BIagp ≥ 5 were defined as the average BIagp based risk thresholds depicting the “Low”, Moderate”, and “High Risk” phases of dengue epidemics, respectively. In the district of Kandy, the average BIagp based risk thresholds was relatively higher as 3.0 ≤ BIagp ≤ 4.1 (Low Risk), 4.2 ≤ BIagp < 5.3 (Moderate Risk), and BIagp ≥ 5.3 (High Risk). In the case of HI, the average thresholds for Colombo district were defined as Low Risk (5.5 ≤ HI ≤ 8.8), Moderate Risk (8.9 ≥ HI < 11.9), and High Risk (HI ≥ 11.9), while 6.9 ≤ HI ≤ 9.0 (Low Risk), 9.1 ≤ HI < 11.8 (Moderate Risk), and HI ≥ 11.8 (High Risk) are recommended for Kandy. These average thresholds are proposed to be applied in other MOH areas located in the district, while the MOH areas investigated in this study could rely upon specific thresholds developed in the current study.Thresholds developed based on BI and HI have been successfully used in dengue epidemic prediction and management in Brazil, Havana, Thailand, Trinidad, and America , 45–47. Ae. aegypti positive breeding sites to the total number of Aedes species positive breeding sites  in Singapore, to investigate the risk of geographical expansion of Ae aegypti. Their findings emphasize that areas with a higher Ae. aegypti BP have a higher degree of geographical expansion of Ae aegypti. Further, such areas tend to have higher case numbers suggesting that the BP of Ae. aegypti is a key factor for the risk characterization of dengue fever in endemic countries, like Sri Lanka [agp that directly reflects larval abundance of Ae. aegypti in the current study.In a recent study, Ong et al.  have useri Lanka . A similagp and HI in Akurana, KMC & GK, and Gampola MOH areas (in the abovesaid previous study) are in close agreement to the threshold values derived by the current ROC approach [In a previous study, a set of entomologically driven thresholds have been developed for a selected set of MOH areas in Kandy, solely based on the natural dynamics of dengue vectors . The curapproach .Ae. aegypti and Ae. albopictus) could exceed 5 (with cases) or 20 (without cases), it could overlook the actual risk for epidemic incidence as Ae. albopictus, which is abundant in the environment, could heavily influence the BI [As a routine practice, the VCE in Sri Lanka utilize BI > 20 (without dengue cases) or BI > 5 (with dengue cases) as the risk threshold for guiding vector control activities. However, these risk levels are often insensitive to the local variations in epidemiological and entomological aspects. Further, even though cumulative BI , along with the physical elimination of breeding sites of locality . ThroughThe absence of records on serological circulations of dengue viruses, limitations in the availability of long term surveillance data  and limited knowledge on herd immunity, is the major limitations encountered in defining the thresholds in the current study . Furtheragp plays a critical role in such circumstances as an early warning signal two-months ahead (two-month lag), providing a valuable window of opportunity for the VCEs to design and initiate vector control activities, before the incidence of a dengue epidemic. This will be a critical determinant in the dengue epidemic management as it can ensure the timely containment of Aedes vectors prior to an outbreak incidence [agp and HI recommended in this study with two months lag period would be of immense importance in responding to dengue epidemics prior to incidence [In Sri Lanka, VCE are struggling to manage dengue epidemics through a variety of approaches such as vector control, environmental, and patient management . Howeverncidence , 25, 48.ncidence , this mancidence . Therefoncidence .In light of this, thresholds defined in this study would be helpful for adopting and directing vector control activities, with optimum utilization of limited human and financial resources to control dengue epidemics , 52. Theagp are significantly associated with dengue epidemics at lag periods of one and two months, emerging as representative stegomyia indices capable of defining dengue epidemics. The BIagp ≤ 2.4 (Low Risk), 3.8 ≤ BIagp < 5 (Moderate Risk), and BIagp ≥ 5 (High Risk) are recommended as the average BIagp based risk thresholds for Colombo, while BIagp ≤ 3.0 (Low Risk), 4.2 ≤ BIagp < 5.3 (Moderate Risk), and BIagp ≥ 5.3 (High Risk) are suggested for Kandy. The average thresholds for HI in Colombo are HI ≤ 5.5 (Low Risk), 8.9 ≥ HI < 11.9 (Moderate Risk), and for Kandy are HI ≤ 6.9 (Low Risk), 9.1 ≤ HI < 11.8 (Moderate Risk), and HI ≥ 11.8 (High Risk). The thresholds derived from this study are recommended to be practiced and validated within the studied areas to suppress dengue vectors before an epidemic, through optimum usage of limited financial and human resources. Furthermore, VCE are recommended to conduct properly planned routine vector surveillance activities to assist the dengue epidemic management based on these thresholds. The present study warrants the essential need in developing entomological thresholds in other areas in the country and use of such prediction approaches in dengue control programmes in Sri Lanka.Based on our findings, both HI and BI


The dawn phenomenon (DP) is the primary cause of difficulty in blood glucose management in type 2 diabetic (T2D) patients, and the use of oral hypoglycemic agents has shown weak efficacy in controlling DP. Thus, this study is aimed at investigating the effect of moderate-intensity aerobic exercise before breakfast on the blood glucose level and glycemic variability in T2D patients with DP.  A total of 20 T2D patients with DP confirmed via continuous glucose monitoring (CGM) participated in the current study. After collecting baseline measurements by CGM as a control, CGM was reinstalled and 30 minutes of moderate-intensity aerobic exercise was performed prior to breakfast. Dawn blood glucose increase, blood glucose levels, and glycemic variability were measured before and after exercise. ΔGlu, 1.25 ± 0.84vs.2.15 ± 1.07, P = 0.005), highest blood glucose value before breakfast , highest blood glucose value before breakfast , highest blood glucose value before breakfast , highest blood glucose value before breakfast , highest blood glucose value before breakfast , highest blood glucose value before breakfast , highest blood glucose value before breakfast , highest blood glucose value before breakfast (Gmax, 8.01 ± 1.16 Dawn blood glucose increase ( Acute moderate-intensity aerobic exercise before breakfast reduced the morning rise of blood glucose in T2D patients, partially counteracting DP. Furthermore, exercise significantly reduced blood glucose fluctuations and improved blood glucose control throughout the day. We recommend that T2D patients with DP take moderate-intensity aerobic exercise before breakfast to improve DP and glycemic control. The “dawn phenomenon” (DP) was first described by Schmidt in 1981 and refers to the spontaneous increase of blood glucose or insulin demand occurring at dawn, despite diabetic patients demonstrating reasonable blood glucose at night without hypoglycemia . While eGlycemic variability is an important indicator of blood glucose control. Specifically, glycemic variability refers to an unstable state in which glucose levels oscillate between peaks and troughs, including short-term blood glucose fluctuations  and long-term blood glucose fluctuations  . GlucoseContinuous glucose monitoring (CGM) systems provide continuous, dynamic blood glucose testing, contributing to the discovery of hyperglycemia and hypoglycemia that are not easily detected by traditional monitoring methods . CGM proExercise plays a crucial role in the integrated glycemia management of patients with type 2 diabetes (T2D). Regular exercise can increase insulin sensitivity, help blood glucose control, and prevent or delay the development of diabetes complications . GuideliPrevious studies have primarily focused on the mechanism, influencing factors, and drug treatment of DP; however, it is not clear whether exercise can lead to improvement. In this study, CGM technology was applied to investigate blood glucose level changes and glycemic variability in T2D patients with DP after moderate-intensity aerobic exercise before breakfast, so as to provide new ideas for the intervention of DP in clinical practice.β-blockers, and individuals who were unable to complete aerobic exercise.A total of 20 T2D patients with DP confirmed via CGM were recruited from the endocrinology department of the China Rehabilitation Research Center between August 2016 and September 2019. All research processes were performed at the center. Subjects were screened in two steps. First, multiple fingertip blood glucose monitoring was conducted for eligible subjects; this was performed at 3 AM along with fasting blood glucose (FBG). When the difference between FBG and nocturnal 3 AM blood glucose level was ≥1.1 mmol/L for three consecutive days, CGM was performed to further screen subjects with DP. All subjects met the diagnostic criteria for T2D defined by the WHO, while DP is defined as the difference between the lowest blood glucose level during the nocturnal period (Gmin) and the highest blood glucose value before breakfast (Gmax) of ≥20 mg/dL (1.1 mmol/L) [This study was approved by the Ethics Committee of China Rehabilitation Research Center; the institutional review board approval number is (2014)K(002). The study conformed to the ethical guidelines of the Declaration of Helsinki. All subjects provided written informed consent before participating.2). Blood pressure was obtained from the right upper arm with a mercurial sphygmomanometer after the subject rested for five minutes. After fasting overnight, each subject received venous blood tests measuring cholesterol (CHO), high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglyceride (TG), and plasma glucose (FPG) by using an automatic analyzer . Fasting insulin (FINS) was measured by a chemiluminescence detector . Glycosylated hemoglobin (HbA1C) was measured with ion-exchange high performance liquid chromatography using an automatic glycohemoglobin analyzer . Insulin resistance levels were assessed via the HOMA-IR homeostasis model calculated by FPG (mmol/L)∗FINS (μIU/L)/22.5. Beta cell function was assessed with HOMA-β and calculated by 20∗FINS (μIU/L)/(FPG (mmol/L) − 3.5) (%) [Clinical information of the subjects was collected at the beginning of the study, including gender, age, height (m), body mass (kg), and body mass index  (%) .ΔGlu) equals Gmax − Gmin. The duration of the DP was defined as the time interval between the nadir and pre-breakfast time points. The blood glucose control level was evaluated by mean blood glucose (MBG), time in range (TIR), mean blood glucose 1 h before breakfast, mean blood glucose after breakfast (within 3 hours), and hyperglycemia area. Intraday glycemic variability was evaluated by blood glucose standard deviation (SD), coefficient of variation (CV), mean amplitude of glucose excursion (MAGE), the largest amplitude of glucose excursion (LAGE), low glycemic index (LBMI), and mean postprandial glucose excursion (MPPGE). Daytime glycemic variability was evaluated using daily differences (MODD).Identical CGM monitors  were used to monitor blood glucose. In order to avoid the deviation caused by the insertion and removal of the sensor, and to reduce the influence of daytime variability, the mean value of two complete 24-hour sessions of recorded data was calculated for subsequent comparison. Subjects were fitted with CGM monitors, and baseline CGM data were collected. After collecting baseline measurements by CGM as control, CGM was reinstalled and exercise was performed. After three days of exercise, the CGM was removed, and the mean value of the two complete 24 hour sessions of recorded data was analyzed again. Indicators, including dawn blood glucose increase, blood glucose levels, and glycemic variability, were measured before and after exercise. The rise of blood glucose at dawn (2max) for three consecutive days. Target heart rate during exercise was equal to heart rate reserve multiplied by training intensity plus resting heart rate . After 3 minutes of exercise, the target heart rate was reached, and subjects continued exercising for 30 minutes at the target heart rate. Additionally, warm-up and cool-down periods of five to ten minutes were performed before and after exercise, respectively. After the third day of exercise, we removed the CGM, analyzed the mean value of the two complete 24 hour sessions of recorded data, and then compared the results with the baseline indicators before exercise.This was a self-controlled study before and after exercise intervention. After baseline CGM data were collected, all subjects were fitted with CGM monitors again and exercised before breakfast (06:27–07:00) on a treadmill  with moderate intensity of 50–60% maximal oxygen uptake . We selected two complete 24 hour sessions of recorded data provided by the CGM for each subject before and after exercise, and then analyzed the mean value of the two complete 24 hour sessions of recorded data to obtain the required indicators. Measurement data were expressed as the mean ± standard deviation (mean ± SD) or median and interquartile range (IQR), as appropriate. This was a self-controlled study before and after exercise. The paired β levels were lower than the normal range.The baseline clinical and laboratory data of the subjects are shown in vs. 8.78 ± 1.09, P = 0.005), ΔGlu , MBG , blood glucose 1 h before breakfast , blood glucose after breakfast , and hyperglycemia area  vs. 0.125 , P = 0.006) were all lower and TIR  was higher than before exercise. By contrast, there was no difference observed in Gmin , CV , MAGE , LAGE , MODD , and MPPGE  were all lower after exercise. However, there was no difference observed in LBMI before and after exercise (Among the glycemic variability indicators, SD (1.1 ± 0.5exercise .According to the criteria recommended by the Chinese Clinical Practice Guideline for continuous glucose monitoring, in which the normal values are MBG < 6.6, SD < 1.4, and MAGE < 3.9 , the MBGβ-cell dysfunction [The pathophysiological mechanism of the DP has not yet been fully established. At dawn, the secretion of insulin antagonistic hormones, such as growth hormone, cortisol, and glucagon, increase. Current research suggests that the DP is the result of the combination of increased endogenous glucose production and persistent insulin resistance in the early morning due to islet function , 18. Abnfunction . The efffunction , 4. DP, function , 19. Thefunction , 4.In this study, we observed that moderate aerobic exercise before breakfast helped to reduce the rise of blood glucose at dawn that often occurs in type 2 diabetes patients. In particular, the significant improvement of blood glucose one hour before breakfast is strong evidence that exercise before breakfast can improve fasting hyperglycemia. Blood glucose was also significantly improved after breakfast, suggesting that exercise before breakfast further improved instances of extended DP. TIR is defined as the proportion of time in which blood glucose is in the range of 3.9 to 10.0 mmol/L during a 24-hour period. CGM technology can accurately obtain TIR. In the International Consensus on CGM Applications in 2017, TIR was recommended as one of the indicators for CGM standardization reports . This stThe acute effects of exercise are beneficial for patients with T2D. Exercise increases the concentration of GLUT-4 in the cell membrane and increases glucose uptake in skeletal muscles –24. Acutβ cells than persistent hyperglycemia [Many studies have shown that fluctuating hyperglycemia more severely damages glycemia , 7, 8. Aglycemia , 7, 34. glycemia . Althougglycemia . This stThe timing of exercise has also become a controversial issue in recent years . The 201In summary, acute moderate-intensity aerobic exercise before breakfast appeared to counteract DP, significantly reduced blood glucose fluctuations, and improved blood glucose control throughout the day. To the best of our knowledge, our research provides the first evidence showing this potential by exercise. This study provides a new concept for T2D patients to ameliorate DP, reduce blood glucose fluctuation, and achieve overall stable blood glucose control. However, whether the observed improvement was due to the simple hypoglycemic effect of exercise or the additional improvement effect of exercise on the mechanism of the DP is not yet clear. Thus, it is necessary to explore the mechanism of improving the DP by further research that considers increasing the number of cases, exercising for a longer duration, and increasing observations of relevant hormone indices.