Chicorium intybus L.) powder and a probiotic blend (PrimaLac®) on productive performance, blood biochemical parameters, and ileal microbiota in broiler chickens. A total of 225 one-day-old broilers (Ross 308) were used in a completely randomized design with five experimental diets as follows: 1—basal-diet without supplements (control-group); 2—basal-diet including probiotic blend; 3— basal-diet including 0.10% chicory; 4—basal-diet including 0.15% chicory; 5—basal-diet including 0.20% chicory. At 42 days of age, representative birds per replicate were randomly selected for blood samples and carcass measurements. Results showed that the body weight gain of broilers fed the probiotic blend or 0.10% chicory was significantly (P < 0.05) higher than those fed on the other treatments. The abdominal fat pad was significantly (P < 0.05) lower in birds fed diets including chicory compared with control or probiotic. Blood triglycerides and LDL levels were reduced (P < 0.05) and HDL increased (P < 0.05) when fed probiotic or chicory whereas no significant effect on the other serum parameters was found. Broiler ileal microflora from the control group had significantly (P < 0.05) higher count of E. coli and lower Lactobacillus than those from the other groups. From findings, it is possible to conclude that dietary chicory powder supported positively growth performance and improved gut microbiota in broiler chickens. However, more research is needed on this subject to better understand the mode of action of feed additives used.The experiment was designed to determine the effect of different levels of chicory ( Antibiotics are usually used in poultry for therapeutic, preventive, and nutritional purposes ,2,3. UsiProbiotics as live microorganisms can also improve the intestinal balance of the host livestock . The mecCicorium intybus L.) is an important medicinally perennial herb plant of the Asteraceae family ; Treatment 3—basal-diet supplemented with chicory powder at 0.10%; Treatment 4—basal-diet supplemented with chicory powder at 0.15%; Treatment 5—basal-diet supplemented with chicory powder 0.20%.A two-phase feeding program was used; a starter diet was used from 1–21 days and a finisher diet from 22–42 days. The ingredients and nutrient composition of diets are shown in Mortality rate and growth performance, as feed intake, feed conversion ratio, and body weight gain, were recorded per pen at 21 and 42 days of age. After 12 h of fasting, five birds per treatment were randomly selected for carcass characteristics measurements at 42 days of age. Birds were sacrificed, plucked, and eviscerated, then the weight of the whole carcass, carcass components (drumsticks and breast), liver, gizzard, and abdominal fat were excised and individually weighed. Carcass traits components were calculated as a percentage of the preslaughter live body weight.g × 10 min to obtain serum. Samples were stored at −20 °C until analyzed for total protein, albumin, glucose, total cholesterol, triglycerides, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and uric acid, using commercial laboratory kits .At the end of the trial (42 days), ten broilers per group (30 birds/treatment) were selected for blood sampling. Blood samples were collected from the wing veins and centrifuged at 3000 Lactobacilli, and Eosin Metilan-Blue  was used for E. coli, then incubated under anaerobic conditions at 37 °C for 72 h. Bacterial units were counted by a colony counter and adjusted to 1 g sample. The bacteria counts were reported as log10 CFU/g of ileal digesta.For the ileal microbial count, at 42 days of age, 30 birds (two birds from each replicate) were randomly selected and killed by cervical dislocation. One gram of ileum content was sampled and transferred into sterile tubes containing 9 ml of sterile PBS and homogenized and serially diluted. A small sample (0.1 ml) from each dilution was inoculated on MRS agar  was used to culture ijk = μ + Ti + Rij + ɛijk, where: Yijk = response variables from each individual replication or pen; µ = the overall mean; Ti = the effect of dietary additive; Rij = the inter-experimental unit (replications) error term; ɛijk = the intra-experimental unit error term. Means were compared for significant differences using the LSMEANS option of SAS/STAT . Statistical significance was established at P < 0.05.The experiment was carried out in a complete randomized design with dietary feed additives as the main effects. The model assumptions of normality and homogeneity of variance were tested using Shapiro–Wilk’s and Levene’s tests, respectively. Data were subjected to analysis of variance (ANOVA) according to the General Linear Model (GLM) procedure of SAS/STAT . The statistical model used was: YE. coli, and in enhancing the growth of beneficial Lactobacillus gut bacteria. Thus, this study demonstrated that natural feed additives can be successfully used as an alternative to antibiotics as growth promoters for broiler chickens.Chicory powder and probiotic blend as natural feed supplements in broiler diet supported positively the growth and feed efficiency of birds, without affecting most of the blood biochemical parameters. Moreover, it is important to underline that probiotics and chicory supplementation showed a significant effect in inhibiting the growth of potentially pathogenic


Poria cocos)-Guizhi (Cinnamomi ramulus) herb pair (FGHP) is a commonly used traditional Chinese herbal formula with coronary heart disease (CHD) treatment potential. However, the mechanism of FGHP in the treatment of CHD was still unclear. In this study, the action targets and underlying mechanism of FGHP against CHD were successfully achieved by combined network pharmacology prediction with experimental verification. 76 common targets were screened out by overlapping the chemical-protein data of FGHP and CHD-related targets. Then, two key targets were further selected for verification by using western blot analysis after analyzing PPI, GO function, and KEGG pathway. Results indicated FGHP could alleviate CHD syndromes and regulate inflammatory responses in acute myocardial ischemia rats, and the reduction of expression of TNF-α and IL-6 in myocardial tissue would be one of its possible underlying mechanisms. Our work demonstrated that network pharmacology combined with experimental verification provides a credible method to elucidate the pharmacological mechanism of FGHP against CHD.The Fuling ( Coronary heart disease (CHD) is a common type of heart disease and one of the most common causes of human deaths in the world , 2. The Poria cocos , Cinnamomi ramulus , Atractylodes macrocephala Koidz. , and Licorice , recorded in the classic ancient book “JinGuiYaoNue” written by Zhongjing Zhang. FGHP has been commonly used clinically for the treatment of CHD. However, the mechanism by which FGHP helps to treat CHD remains unclear, which impedes its further clinical application and spread in some degree.Traditional Chinese medicine (TCM), mainly Chinese herbal medicine, has been used for thousands of years to treat CHD and its related diseases, as it is characterized by multiple components, and it can treat diseases through multiple pathways and targets , 9. HerbNetwork pharmacology recently has become a new and a powerful approach to systematically reveal the principles and function of a complex biological system along with the rapid progress of bioinformatics , 12, andTherefore, network pharmacology-based study was adopted in this study to investigate underlying action mechanism of FGHP in CHD. First, the potential action of FGHP was predicted using a network pharmacological method. Subsequently, an isoproterenol- (ISO-) induced acute myocardial ischemia (AMI) experiment was carried out on rats to confirm the network analysis-based data.http://www.nrc.ac.cn: 9090/ETCM), the Traditional Chinese Medicine Integrated Database , and the Traditional Chinese Medicine Systems Pharmacology Database . The active components were the filtered by combining oral bioavailability (OB) ≥30% and drug-likeness (DL) index ≥0.18 as suggested by the TCMSP database.Firstly, chemical compounds in Fuling (FL) and Guizhi (GZ) were obtained from the Encyclopedia of Traditional Chinese Medicine  and STITCH (http://stitch.embl.de/) databases were used to identify the potential human protein targets of FGHP's active compounds. At the same time, CHD-related human genes were downloaded from OMIM (https://omim.org/) and DisGeNET (https://www.disgenet.org/) databases. Then, the candidate targets of FGHP against CHD were obtained by overlapping the above targets with a Venn diagram (http://bioinfogp.cnb.csic.es/tools/venny/index.html).SWISS (https://david.ncifcrf.gov/) to obtain the related functions and pathways.The gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the candidate targets were carried out using the DAVID (https://string-db.org/) database, which supplies information regarding the predicted and verified experimental interactions of proteins [Homo sapiens” and interaction score >0.7 (high confidence). The top 10 proteins with higher levels of correlation were collected as the center targets for FGHP against CHD.The protein-protein interaction (PPI) data were gathered from STRING , which is a software package for visualizing network analysis [Based on these results, a compound-target-pathway network composed of compounds, corresponding targets, and their related potential pathways was constructed and displayed using Cytoscape 3.7.1  were obtained from the Hubei Experiment Animal Research Center . All rats were housed in the SPF laboratory animal room, Hubei University of Chinese Medicine , and maintained in suitable conditions with a 12 h light/dark cycle and room temperature at 22 ± 2°C and 50 ± 10% humidity. They had free access to food and water. The study protocol was approved by Animal Ethics Committee of Hubei University of Chinese Medicine (HUCMS-201903001). Fuling and Guizhi were purchased from TCM pharmacy of Wuhan Hospital of Traditional Chinese Medicine . The FGHP was obtained by mixing the two herbs (Fuling and Guizhi) at a mass ratio of 1 : 1 and then grinding them into powder. Propranolol tablets were purchased from Ya Bang Pharmaceutical Factory . Isoproterenol (ISO) hydrochloride was purchased from Southwest Pharmaceutical Co., Ltd. .n = 6 per group): ISO, ISO + FGHP (1.8 g/kg), and ISO + Propranolol . The rats in the two ISO+ groups were pretreated by oral gavage administration with propranolol or FGHP, while the rats in the control group and the ISO group were pretreated by oral gavage administration with equal volumes of 0.9% saline once a day for 2 weeks. Afterwards, the rats in all three administration groups were subcutaneously injected with ISO (85 mg/kg) for two consecutive days [Rats were randomly divided into the control group and three administration groups  were recorded under pentobarbital anesthesia using a BL-420F biological data acquisition and analysis system. Blood was collected via the abdominal aorta. The serum was separated by centrifugation at 4°C and 3,000 rpm for 15 min and subsequently stored at −20°C for biochemical analysis. The heart tissues were dissected and immediately frozen in liquid nitrogen or fixed in 10% paraformaldehyde for western blot or histological analysis.The serum creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) levels were measured by using commercial kits and following the product instructions .The myocardial tissues were dissected and then fixed in 10% paraformaldehyde overnight followed by processing of dehydrating, paraffin embedding, sectioning, and staining (H&E). Morphological structure of each specimen was examined and photographed by using a light microscope .α (#6954) , and anti-β-actin  at 1 : 1000 dilution at 4°C overnight. Following washing in TBST, the membranes were incubated with corresponding horseradish peroxidase-conjugated secondary antibodies. Then, the protein bands were visualized with a BeyoECL Plus . Densitometry analysis of each band was performed using the BioRad Quantity One software. β-Actin protein was used as the internal control for semiquantitative analysis.The total protein from the myocardial tissues were extracted as described previously. Next, tt-test was used to analyze statistical comparisons between two groups. Multiple comparisons were assessed by one-way ANOVA. P < 0.05 was assumed as statistically significant.Data were expressed as the mean ± SEM. GraphPad Prism 5 software  was used for the statistical analysis and graphics. Unpaired Totally, 33 components of Fuling (FL) and 220 components of Guizhi (GZ) were obtained from the ETCM, TCMID, and TCMSP . All actA total of 912 CHD-related targets were obtained from the DisGeNET and OMIM. The detailed information of the targets is shown in P < 0.05, To explore the multiple mechanisms of FGHP on CHD from a systematic level, gene ontology (GO) analysis and KEGG pathway enrichment of the 76 candidate targets were performed. In total, 96 enriched GO terms and 48 pathways were identified , matrix metalloproteinase 9 (MMP9), mitogen-activated protein kinase 8(MAPK8), mitogen-activated protein kinase 3(MAPK3), interleukin 6 (IL6), epidermal growth factor receptor (EGFR), vascular endothelial growth factor A (VEGFA), tumor necrosis factor (TNF), retinoid X receptor alpha (RXRA), and prostaglandin-endoperoxide synthase 2 (PTGS2).Based on the all above information, we constructed a compound-target-pathway network  to explaAs shown in P < 0.01). Pretreatment with FGHP and propranolol markedly inhibited the elevation in the levels of serum CK-MB and LDH of the ISO group (P < 0.01) as shown in Figures The serum concentrations of creatine kinase‐MB (CK-MB) and lactic dehydrogenase (LDH) in the ISO group were significantly increased than those in the control group  for experimental validation. As shown in α and IL-6 were significantly increased in the ISO group compared with those in the control group (P < 0.01). Pretreatment with FGHP or propranolol showed that TNF-α and IL-6 expression decreased obviously in the ISO + FGHP or the ISO + Pro groups compared with the ISO group (P < 0.05).Based on the network pharmacology analysis, to verify the results derived using the PPI and compound-target-pathway network, we chose the high two related targets  in lipopolysaccharide- (LPS-) induced cardiomyocyte injury model rats [α and IL-6 to exert their anti-inflammatory properties [α and IL-6 were selected as the evaluation indicators to observe the therapeutic effect and possible mechanism of FGHP in ISO-induced AMI rats.FGHP originated from Linggui Zhugan Decoction, which has actually been medically used for lots of years in the treatment of several diseases, including cardiovascular diseases and metabolic diseases . It was del rats . Lingguidel rats . More imoperties , 37. So,in vivo studies, we attempted to study the therapeutic effect of FGHP and confirm the predicted results from network pharmacology. To this end, we tested this agent in rats with ISO-induced AMI. Our results showed that characteristic indices including CK-MB and LDH content were decreased significantly in FGHP pretreated rats compared with AMI control rats (shown in In shown in . In addishown in  and myocshown in . These rα and IL-6 expressions were detected in myocardial tissues in the ISO group rats; however, both of them were significantly reduced with FGHP pretreatment (shown in α and IL-6 activity to modulate inflammation is at least in part responsible for FGHP exerting its cardiac protective efficacy. Further molecular mechanism studies and confirmation of the prediction of FGHP are required.Moreover, we further verify the pharmacological mechanism of FGHP on treatment for CHD by measuring the expression levels of the core and specific targets. Upregulated TNF-shown in . These fshown in , indicatα and IL-6 for the suppression of inflammatory response in cardiac muscle to exert curative effect for CHD. The results partially explained the pharmacological mechanism of FGHP against CHD.In summary, our study predicted and verified that FGHP could reduce protein expression of TNF-


Omadacycline is a once-daily oral or intravenous (i.v.) aminomethylcycline antibiotic approved in the United States for the treatment of community-acquired bacterial pneumonia (CABP) and acute bacterial skin and skin structure infections (ABSSSI) in adults. Omadacycline pharmacokinetics were characterized in 18 patients with hepatic impairment and 12 matched healthy subjects. Patients with hepatic impairment received i.v. omadacycline at 100 mg (mild hepatic impairment) or 50 mg (moderate and severe hepatic impairment) and oral omadacycline at 300 mg (mild hepatic impairment) or 150 mg (moderate hepatic impairment); oral omadacycline was not evaluated in those with severe hepatic impairment. Omadacycline is a once-daily oral or intravenous (i.v.) aminomethylcycline antibiotic approved in the United States for the treatment of community-acquired bacterial pneumonia (CABP) and acute bacterial skin and skin structure infections (ABSSSI) in adults. Omadacycline pharmacokinetics were characterized in 18 patients with hepatic impairment and 12 matched healthy subjects. Patients with hepatic impairment received i.v. omadacycline at 100 mg (mild hepatic impairment) or 50 mg (moderate and severe hepatic impairment) and oral omadacycline at 300 mg (mild hepatic impairment) or 150 mg (moderate hepatic impairment); oral omadacycline was not evaluated in those with severe hepatic impairment. Safety monitoring included the collection of adverse events (AEs), performance of laboratory tests, determination of vital signs, and performance of electrocardiograms. Omadacycline exposures were similar in patients with hepatic impairment and healthy subjects following i.v. or oral administration, with the geometric mean ratios for the area under the concentration-time curve and the maximum drug concentration ranging from 0.79 to 1.42. Omadacycline was safe and well tolerated. Overall, 13/30 (43.3%) participants experienced an AE; those occurring in more than 1 participant included headache (13.3%), nausea (6.7%), infusion-site pain (6.7%), contusion (6.7%), and dizziness (6.7%), with no differences based on the degree of hepatic impairment or the route of administration. Asymptomatic increases in heart rate were observed; none was considered an AE. These findings suggest that no omadacycline dose adjustment is warranted in patients with hepatic impairment. Contemporary antibiotics often require dose adjustment for patients with some degree of hepatic impairment in order to maintain exposure largely within the range expected for the same dose given to patients who do not have hepatic impairment. Tigecycline, a glycylcycline tetracycline derivative, requires a dosage reduction by 50% in patients with severe hepatic impairment . Intravet1/2) of approximately 17 h, thus supporting once-daily dosing  and acute bacterial skin and skin structure infections (ABSSSI) . Omadacyy dosing . Oral biy dosing . Proteiny dosing . In phasy dosing –14. Nonc dosing –. Clinica dosing –. The purEighteen patients with mild (group 1), moderate (group 2), or severe (group 3) hepatic impairment and 12 healthy subjects were enrolled. Hepatic impairment was classified according to the Child-Turcotte-Pugh scoring method . HealthyTmax) ranged from 0.25 to 0.5 h with i.v. doses and from 1.5 to 2.0 h with oral doses. Similar clearance was observed across the groups: the mean total body clearance following i.v. administration (CL) ranged from 9.1 to 14.2 liters/h, and the mean apparent total body clearance following oral administration (CL/F) ranged from 48.4 to 72.1 liters/h. The terminal elimination half-life (t1/2) was relatively consistent and ranged from 8 to 16 h across the groups, with no apparent trend relative to the degree of hepatic impairment.A comparison of plasma PK parameters for each group showed the expected dose-related differences in exposure for both the i.v. and oral routes of administration . Generalinf), the area under the concentration-time curve from time zero to the last quantifiable concentration point (AUClast), and the maximum drug concentration (Cmax) in patients with hepatic impairment were comparable to those in healthy subjects  between exposure to omadacycline and the severity of hepatic impairment  participants experienced at least 1 AE. AE rates were similar between patients with hepatic impairment and healthy subjects. The most commonly reported AEs were headache (13.3%), nausea (6.7%), infusion-site pain (6.7%), contusion (6.7%), and dizziness (6.7%). All other AEs were reported in only 1 participant. One patient, a 47-year-old male with moderate hepatic impairment, experienced serious AEs of alcohol intoxication with angina pectoris, hypocalcemia, hypotension, and rhabdomyolysis; these occurred 8 days after receiving oral omadacycline at 150 mg. He had previously received i.v. omadacycline at 50 mg, which he tolerated well. This patient had a history of alcoholic cirrhosis, hepatitis C, hypertension, and diabetes mellitus, among other conditions. The events resolved after supportive care in the hospital; none of these events was considered related to omadacycline. One patient with mild hepatic impairment discontinued the study due to a mild facial rash, which occurred several hours after receiving i.v. omadacycline at 100 mg. This event was considered related to omadacycline; the rash was treated with diphenhydramine and resolved after approximately 4 days. Other than the laboratory findings in the patient with the serious AEs described above, there were no clinically relevant changes in serum chemistry, hematology, or physical examination findings during the study.Participants in all treatment groups experienced a mean maximum increase in heart rate (from predose to any postdose measurement) that ranged from 3 to 19 beats per minute. Mean maximum increases in systolic and diastolic blood pressure ranged from 8 to 13 mm Hg and 3 to 10 mm Hg, respectively. Increases in heart rate and blood pressure were transient; they typically occurred within 6 to 12 h after dosing, returned to normal levels by 24 h after the dose, and were not reported as AEs. Electrocardiogram (ECG) data reflected the heart rate changes but showed no other notable findings. There were no apparent differences in these effects between the patients with hepatic impairment and the healthy subjects; heart rate changes tended to occur earlier following i.v. administration than following oral administration.Cmax across all study groups after i.v. administration showed no association between exposure to omadacycline and the Child-Turcotte-Pugh score. Different degrees of hepatic impairment had no clinically relevant effect on the systemic clearance of i.v. omadacycline.Omadacycline is incompletely absorbed from the gastrointestinal tract, resulting in an oral bioavailability of about 35%. Systemically available omadacycline undergoes negligible biotransformation and is eliminated largely unchanged in feces and urine . Followiin vitro not to be a substrate of several known biliary and renal transporters, such as OATP1B1, OATP1B3, MRP2, and BCRP . Modest, asymptomatic increases in heart rate were observed for several hours after dose administration, but no increase was considered to be an AE or associated with ECG abnormalities. Asymptomatic increases in heart rate have also been observed in other phase I clinical studies of omadacycline , 21, whiA possible limitation of this study is that oral omadacycline was not administered to patients with severe hepatic impairment. If a drug is to be administered by more than 1 route, only the route that provides the most information about the impact of hepatic impairment on the drug’s elimination needs to be studied . OmadacyOlder antibiotics in the tetracycline class  have generally not been studied or specifically labeled for use in patients with hepatic impairment –24. Data–With respect to nontetracycline antibiotics, reduced doses of i.v. lefamulin  are required in patients with severe hepatic impairment . LefamulThe potential for hepatotoxicity is well documented with some antibiotics, including tetracyclines, clavulanic acid, macrolides, sulfonamides, and linezolid; however, hepatotoxicity is rare for most other classes of antibiotics . The risThe availability of once-daily oral and i.v. dosing and the lack of a need for dosage adjustment with omadacycline in populations with liver disease offer simplicity and possible advantages for its use in the treatment of common bacterial infectious diseases.The study was conducted at 2 centers in the United States  according to the ethical principles of the Declaration of Helsinki and good clinical practice. The study protocol, consent form, and all amendments were reviewed and approved by an independent ethics committee or institutional review board for each center . Written informed consent was obtained from each study participant prior to enrollment.This was an open-label, fixed-sequence study in adult patients with hepatic impairment and healthy adult subjects. The degree of hepatic impairment was classified according to the Child-Turcotte-Pugh scoring method . Two sepThe study consisted of a screening period that did not exceed 28 days, a baseline period, and 2 treatment periods. During period 1, patients with mild hepatic impairment (group 1) received a single i.v. dose of omadacycline at 100 mg, and those with moderate and severe hepatic impairment (groups 2 and 3) received a single i.v. dose of omadacycline at 50 mg; omadacycline was administered as a 30-min i.v. infusion. A washout period of at least 7 days separated periods 1 and 2. During period 2, groups 1 and 2 received a single oral dose of omadacycline at 300 mg or 150 mg, respectively; they were required to fast for 10 h before and 4 h after the dose. In both periods, healthy subjects matched to groups 1 and 2 received the same doses as the subjects in their respective hepatic impairment groups . An end-2), had normal vital signs with no orthostatic changes, and were in good health generally. Patients with hepatic impairment had to meet the same criteria for age, weight, and vital signs; they also had to have a Child-Turcotte-Pugh score of at least 5 and to have been in a stable condition for at least 3 months prior to enrollment. In all groups, the following exclusion criteria were applied: tobacco use (>10 cigarettes per day), recent blood donation or a hemoglobin concentration of <10 g/dl, a history of hypersensitivity to omadacycline or similar drugs (tetracyclines), a history of malignancy, and a history of any medical condition that could interfere with the conduct of the study. Pregnant or lactating women were excluded, and women of childbearing potential had to use an acceptable form of contraception. Healthy control subjects were also excluded for the following reasons: clinically significant ECG abnormalities; a history or presence of impaired renal function; and the use of other prescription medications, herbal supplements, over-the-counter drugs, or investigational drugs. Patients with hepatic impairment were also excluded for a calculated creatinine clearance of <50 ml/min or the use of investigational drugs; these patients could receive their routine prescription or over-the-counter medications, but drugs such as antacids, calcium-containing supplements, sucralfate, lactulose, and binding resins could not be taken for at least 12 h before and after the administration of omadacycline.Healthy control subjects were eligible if they were aged 18 to 70 years, weighed at least 50 kg , obtained by direct venipuncture or an indwelling cannula inserted in a forearm vein, were collected prior to dose administration and at 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, 24, 48, 72, and 96 h after dose administration in tubes containing sodium heparin. The samples were centrifuged for 10 min within 30 min of collection. The resulting plasma was transferred to polypropylene screw-cap tubes and placed on dry ice. The tubes were kept frozen at −70°C or colder until analysis. The plasma concentrations of omadacycline were measured using a validated liquid chromatography-mass spectrometry/mass spectrometry assay having a lower limit of quantitation of approximately 20 ng/ml. Calibration standard responses were linear over the range of 20 to 2,000 ng/ml. In plasma, the interday assay accuracy, expressed as percent relative error for quality control (QC) concentrations , ranged from −5.3% to 2.0% bias in QC samples. Assay precision, expressed as the interday percent coefficients of variation of the mean estimated concentrations of the QC samples, ranged from 3.1% to 4.5%. The extraction efficiency ranged from 87.0% to 93.5% across the QC samples. The overall mean matrix effect at 60, 758, and 1,520 ng/ml was 1.8%.At screening and baseline (the day prior to dosing), the study participants underwent a physical examination; determination of vital signs , height (at screening only), and body weight; a serum or urine pregnancy test; and tests for alcohol and drugs. Blood pressure and heart rate were measured at screening, baseline, and predose, as well as at multiple time points on the day of dosing and at the end-of-study visit. A urinalysis was performed at screening, as were hepatitis virus and HIV tests. A 12-lead ECG was obtained at screening and baseline; at 2, 12, and 24 h after dosing; and at the end-of-study visit. Serum chemistry and hematology testing were carried out at screening and baseline, at 24 and 48 h after dosing, and at the end-of-study visit. Safety assessments consisted of collecting all AEs and serious AEs, noting their severity and relationship to the study drug.last and AUCinf, each of which was calculated by the linear trapezoidal linear interpolation method; Cmax; CL; apparent total body clearance following oral administration (CL/F); apparent volume of distribution (beta method) following i.v. administration (zV); apparent volume of distribution (beta method) following oral administration (zV/F); Tmax; and t1/2, calculated from the PK program that included a minimum of 3 terminal time points.The plasma concentrations of omadacycline are reported in nanograms per milliliter. Except for the predose samples, concentrations below the limit of quantitation were excluded from the calculation of the PK parameters. Plasma concentration-time data were analyzed using standard noncompartmental methods incorporating SAS programs validated with WinNonlin software . PK parameters included AUClast, AUCinf, and Cmax. Secondary variables were Tmax, CL or CL/F, zV or zV/F, and t1/2. The numerical Child-Turcotte-Pugh score  and a random effect for matching block or impaired subject/healthy subject pair. The 90% confidence intervals were calculated for the ratio of PK parameters for impaired versus healthy subjects. All analyses were done on the logarithmic scale and back-transformed for reporting. Analysis was performed separately for the i.v. and oral dose parameters. Dose-normalized AUC values after a single i.v. infusion were pooled across all study groups, and the relationship to the Child-Turcotte-Pugh score was investigated by exploratory regression. Various linear and polynomial regressions were explored to find a well-fitting statistical model for this relationship.AUC and medinfo@paratekpharma.com for review.Paratek Pharmaceuticals, Inc., has a commitment to ensure that access to clinical trial data is available to regulators, researchers, and trial participants, when permitted, feasible, and appropriate. Requests for deidentified patient-level data may be submitted to


Clinical studies of new antitubercular drugs are costly and time-consuming. Owing to the extensive tuberculosis (TB) treatment periods, the ability to identify drug candidates based on their predicted clinical efficacy is vital to accelerate the pipeline of new therapies. Recent failures of preclinical models in predicting the activity of fluoroquinolones underline the importance of developing new and more robust predictive tools that will optimize the design of future trials. Clinical studies of new antitubercular drugs are costly and time-consuming. Owing to the extensive tuberculosis (TB) treatment periods, the ability to identify drug candidates based on their predicted clinical efficacy is vital to accelerate the pipeline of new therapies. Recent failures of preclinical models in predicting the activity of fluoroquinolones underline the importance of developing new and more robust predictive tools that will optimize the design of future trials. Here, we used high-content imaging screening and pharmacodynamic intracellular (PDi) modeling to identify and prioritize fluoroquinolones for TB treatment. In a set of studies designed to validate this approach, we show moxifloxacin to be the most effective fluoroquinolone, and PDi modeling-based Monte Carlo simulations accurately predict negative culture conversion (sputum sterilization) rates compared to eight independent clinical trials. In addition, PDi-based simulations were used to predict the risk of relapse. Our analyses show that the duration of treatment following culture conversion can be used to predict the relapse rate. These data further support that PDi-based modeling offers a much-needed decision-making tool for the TB drug development pipeline. Mycobacterium tuberculosis is the leading cause of death from a single infectious agent. The so-called short course treatment of drug-susceptible TB   caused by and INH)  remains ctivity 26. They activity 2. HoweverIt was anticipated that by introducing fluoroquinolones into drug-susceptible regimens, the treatment period could be reduced by 2 months (from 6 to 4 months) . Clinicain vivo results (Pharmacokinetic/pharmacodynamic (PK/PD) models are useful in evaluating the length of treatment required by new regimens of anti-TB drugs. Recent clinical trials have highlighted the concerns of preclinical studies. Animal models show dynamics that differ from those observed in humans , 15, and16– results . However results , 21.in vitro intracellular (macrophage) M. tuberculosis high-content imaging-based platform, termed intracellular pharmacodynamics (PDi), can be a powerful tool for predicting the activity of first-line TB drugs in patients . The intracellular anti-tubercular activity of MXF, levofloxacin (LVX), norfloxacin (NOX), ofloxacin (OFX), sparfloxacin (SPX), and ciprofloxacin (CIP) was assessed at a concentration ranges between 0.01 and 100 mg/liter. The kill rate elicited by each concentration was then calculated as previously described (50) and the maximal kill rates for each drug .The efficacy of selected fluoroquinolones against intracellular escribed . Figure escribed . This moach drug  to f. Fi50 (0.051 mg/liter), followed by MXF, CIP, and LVX , while OFX and NOX displayed poor activity with EC50s of 1.414 to 1.705 mg/liter, as shown in 50s, the maximal kill rate for all fluoroquinolones was determined to be comparable  M. tuberculosis , the kill rate/growth rate ratio of MXF at a maximal concentration (100 mg/liter) was 1.82, similar to that of RIF at a maximal concentration  .To compare our predictions with the clinical literature, we used Monte Carlo simulations to generate hypothetical treatment outcomes when using the same regimens previously used in clinical trials. PDi modeling predicted that within 8 weeks 96% of the patients would achieve culture-negative status when INH or ETB is replaced with 400 mg of MXF (the PK/PD properties of partner drugs are based on reference Emax) of MXF is the most influential parameter upon bacillary clearance. The second most significant parameter is the initial intracellular burden the patient presents at the clinic, concurring with previous clinical studies (50). The potency and PK parameters of RIF ranked lower than MXF parameters in our sensitivity analysis. The parameters for other partner drugs  played a negligible role in the overall outcome of the simulations.Table S3 displays the sensitivity analysis of the PDi modeling , which indicates that the maximal kill rate , we compared the overall PK properties and the ELF exposures of MXF, CIP, and LVX (Table S4). Simulations predict MXF to have superior PK properties due to its relatively high AUC level in the plasma, and it displays the highest accumulation in the ELF. Table S4 shows that LVX at a higher dose of 750 mg/day would achieve a pulmonary PK profile very similar to that observed for MXF at 400 mg/day. Assuming linear PK for LVX, 500 mg/day is expected to achieve ELF exposure inferior to 400 mg/day of MXF. At a 1,000-mg dose, LVX has a comparable ELF AUC value (221.7 mg ⋅ h/liter) compared to a standard dose of MXF of 400 mg/day (173.1 mg ⋅ h/liter). Systematic PK parameters were chosen from previously reported data (per Table S4).Simulating various doses of LVX and MXF and integrating this with our imaging-based PDi data reveals that an increase of LVX from 500- to 1,000-mg results in a remarkable improvement in activity . This allows for the identification of the best candidates for further analysis. The results demonstrated that four of the six fluoroquinolones have similar potencies . Despite RIF, our control compound, being superior overall, MXF displayed a similar maximal kill rate in both the live fluorimetry and the fixed Operetta assay. However, patients treated with a 4-month regimen containing MXF showed higher rates of disease relapse despite achieving faster culture conversion in comparison to standard treatments . MXF, ho50, cytotoxicity was observed, as reported in the literature . Figure S4 displays the biexponential nature of CFU reduction which, in our simulation, is a direct result of having two separate populations (extracellular and intracellular) .Similar to the work described previously , we showM. tuberculosis killing) is able to predict long-term clinical responses is perhaps not surprising when it is considered that the majority of patients (ca. 80%) culture convert within ca. 8 weeks of treatment  M. tuberculosis , levofloxacin (LVX), norfloxacin (NOX), ofloxacin (OFX), sparfloxacin (SPX), and ciprofloxacin (CIP) were purchased from Sigma, UK. All compounds were made up in dimethyl sulfoxide (Sigma).M. tuberculosis H37Rv expressing the far-red reporter mCherry was used in this study (H37Rv-mCherry) , and 10% heat-inactivated fetal bovine serum  at 37°C and 5% CO2.mCherry) . Aliquot5 cells per well, and differentiated for 72 h in supplemented RPMI 1640 and 100 ng/ml phorbol 12-myristate 13-acetate  at 37°C and 5% CO2. Differentiated THP-1 cells were infected with H37Rv-mCherry in suspension at a multiplicity of infection of 1:5 in FluoroBrite Dulbecco modified Eagle medium (DMEM) supplemented with 10% HI-FBS and l-glutamine for 24 h at 37°C. For optimal macrophage environment, each 96-well plate was covered with a Breath-EASIER sealing membrane (Sigma) to allow gaseous exchange but maintaining containment level 3 safety regulations. This has been refined from our previous work, improving the THP-1 cell growth conditions. After 24 h, the cells were washed, and the drugs at the required concentrations were added in FluoroBrite DMEM to a total volume of 200 μl. Infected cells were incubated for up to 144 h.THP-1 cells were differentiated in Perkin-Elmer CellCarrier-96 plates, seeded at 5 × 10M. tuberculosis kill kinetics were obtained by incubating M. tuberculosis H37Rv in the presence of test drug, followed by plating to obtain CFU. M. tuberculosis was cultured at 37°C in Middlebrook 7H9 broth (Difco) supplemented with 0.05% (vol/vol) Tween 80 (Sigma), 0.2% (vol/vol) glycerol, 10% OADC (7H9), and 50 mg/liter hygromycin (Sigma) with magnetic stirrers to the mid-log-growth phase before 2-fold dilutions of test drug ranging from 15,360 to 30 ng/ml were added. A sample of culture was plated at 0 h before drug was added to obtain the initial bacterial count. After the addition of drug, 2-ml aliquots of bacterial culture with magnetic stirrers were incubated at 37°C in complete media. At defined time intervals of 24, 48, 72, 96, and 168 h, aliquots were pelleted to remove drug. These were serially diluted in PBS, plated on Middlebrook 7H11 agar, and incubated at 37°C in 5% CO2 for 28 days. The Miles Misra method was used to determine CFU (Extracellular-grown (planktonic) mine CFU .n ≥ 3), all performed at least in triplicate, and data were produced via two methods. First, each plate was screened every 24 h for fluorescence using a Varioskan  at an excitation of 578 nm and emission of 610 nm, thus producing “live” fluorometer readouts. In addition, plates were fixed with 5% paraformaldehyde (Sigma) for 2 h for imaging using an Operetta (Perkin-Elmer) with a 60× High NA objective, as described previously . Hence, the model assumes that there are no positive or negative PD interactions between the drugs  and kill rate (0.23h−1) values were derived in this study . The LVX extracellular Emax and EC50 values were assumed to be similar to those for MXF for the purposes of this work.Extracellular-grown us study , whereasParameters for RIF, INH, PZA, and EMB were derived from our previous work , whereasM. tuberculosis growth rates , including those reported by us, range from a DT of 21 h to a DT of 48 h . Extracellular and intracellular bacilli respond differently to drug treatment, and both reservoirs were simultaneously simulated for each patient, resulting in a biexponential decrease in total CFU burden over time . Overall, the drug kill rate  is equal to the rate of the fastest-acting drug at the same time. This is determined by the epithelial lining fluid (ELF) drug exposure and its intrinsic rate of kill (as defined in vitro) at any given time point for each drug.The models utilized for Monte Carlo simulations here were as described in detail elsewhere , 31. Bri7 CFU/ml (C in equation 2 represents the drug concentration in the ELF); ELF levels were estimated by using the ELF/serum ratio reported in the literature, as previously described for RIF, ETB, INH, and PZA, and an ELF ratio of 5.2 was estimated for MXF . Data generated form Monte Carlo simulation from different scenarios were compared head to head, and the odds ratio with 95% CI was estimated accordingly.Sensitivity analysis was performed to assess the most influential parameters upon the bacillary clearance within the simulation (Table S3). The analyses were performed using the FME package  with R version 3.4.2 . InfluenM. tuberculosis can effectively measure bacillary growth, this approach has a limited dynamic range when assessing bacillary sterilization over time when measurements fall below the minimum limit of fluorescence detection. To overcome this issue, high-content (Operetta) imaging was used to more accurately measure time-dependent bacillary sterilization for specific drugs. With regard to intracellular M. tuberculosis growth, during the course of this and our other studies, as well as in studies by other laboratories, we noted a variation in intracellular M. tuberculosis DT, as mention above under “Modeling Parameters.” To normalize for this in our mathematical modeling, we selected a DT of 21 h throughout. Furthermore, for the purpose of the mathematical modeling/simulations to predict clinical outcome, in the absence of evidence to the contrary, we also made a number of assumptions. First, it was assumed that there is no pharmacological interaction (synergy/additivity/antagonism) between the modeled drugs; therefore, the clinically observed M. tuberculosis sterilization rate was equivalent to the sterilization rate of the drug with the fastest sterilization rate. Second, it was also hypothesized that PD interactions of drugs against intracellular (macrophage) M. tuberculosis measured in in vitro culture were similar to those in the ELF.While fluorometry-based measurements of intracellular M. tuberculosis sterilization rates and PDi PK modeling have been used in this study to predict the clinical activity of fluoroquinolones against TB, this approach generates dynamic parameters  that are not suitable for comparison with traditional static microbiological parameters such as the MIC.It should be noted that while time-dependent intracellular


To investigate the association between high-normal preconception TSH levels and reproductive outcomes in infertile women undergoing the first fresh D3 embryo transfer.  This was a retrospective study. Euthyroid patients undergoing the first fresh D3 embryo transfer from January 2018 to May 2019 were initially included. The patients were divided into a low-TSH (0.27–2.5 mIU/L) group and a high-normal TSH (2.5–4.2 Miu) group. The reproductive outcomes were compared between the groups. P = 0.416, 0.163, and 0.219). No significant differences were observed in terms of gestational age, single live birth rates, and birth weight, or birth length.  A total of 1786 women were ultimately included, in which 1008 of whom had serum TSH levels between 0.27 and 2.5 mIU/L and 778 of whom had serum TSH levels between 2.5 and 4.2 mIU/L. The patients were highly homogeneous in terms of general characteristics. High-normal TSH levels had no adverse impact on the clinical pregnancy rate, miscarriage rate, or live birth rate (respectively, aOR = 0.92, 1.30, and 0.88 and  High-normal TSH levels did not significantly influence reproductive outcomes in infertile women undergoing the first fresh D3 embryo transfer. Further studies are needed to test whether the results might be applicable to a wider population. Subclinical hypothyroidism (SCH), which was common in clinical practise, was a subtle deficiency in thyroid function regardless of the elevated thyrotropin (TSH) level and normal free thyroxine FT4) level . There i level . A total of 2542 infertile women receiving fresh D3 embryo transfer in the Department of Reproductive Medicine, the Third Affiliated Hospital of Zhengzhou University, from January 2018 to May 2019, were initially included. The women underwent standard controlled ovarian hyperstimulation (COH) with a downregulation protocol using a GnRH-agonist or an antagonist protocol according to the patients' conditions. For women with poor ovarian response, mild ovarian stimulation was offered. Oocytes were retrieved transvaginally 34–36 h after HCG administration and were inseminated using either the IVF or the ICSI method. Rescued ICSI was performed when necessary to avoid failure of insemination. Variable cleavage embryos were observed and scored accordingly on the third day after insemination. High-quality embryos were preferentially transferred to the uterus under the guidance of ultrasound.n = 361); the presence of overt or subclinical thyroid dysfunction (n = 366); and lack of essential data (n = 29). After exclusions, 1786 women were transferred with fresh D3 embryos at the first IVF/ICSI cycle and were divided into two groups based on preconception TSH levels . As euthyroid women with preconception TSH<4.2 mIU/L were included in this study, no further tests or treatments were provided to them after starting the ovarian stimulation. The study has been carried out in accordance with the Code of Ethics of the Declaration of Helsinki.Patients with the following criteria were excluded: previous embryo transfers . Clinical pregnancy was defined as the presence of the gestational sac and fetal heart activity following positive serum β-hCG. Early embryo loss was defined as the lack of visible gestational sac or fetal heart activity following positive serum β-hCG. Ectopic pregnancy was defined as a pregnancy that did not occur inside the uterine cavity. Miscarriage, which included early abortion, intermediate abortion, and late abortion, was defined as a pregnancy that did not result in delivery. Preterm birth was defined as a gestational age less than 259 days (37 weeks). Live birth was defined as the delivery of live babies. Low live birth weight was defined as the neonatal weight less than 2500 g.Pregnancy was defined as positive serum t test or the Mann–Whitney U test where appropriate. Categorical variables were expressed as numbers or percentages, and the chi-squared test or Fisher's exact test was used for comparisons of categorical variables where appropriate. Binary logistic regression was then used to evaluate the impact of preconception high-normal TSH levels on the clinical and obstetric outcomes. Data were analyzed using SPSS 22.0 statistical software. All P values less than 0.05 were considered statistically significant.Our primary outcomes were clinical outcomes, such as clinical pregnancy rate, early pregnancy loss rate, miscarriage rate, and live birth rate. The secondary outcomes were obstetric outcomes, such as the single live birth rate, birth weight, birth length, and gestational age. Continuous data are expressed as the mean (SD), and statistical comparisons were performed by either Student's P = 0.107, 0.126). The frequencies of insemination methods and number and high-quality of transferred embryos were similar in both the groups . No difference was noted in terms of the scheme of ovarian stimulation (P = 0.355).The flowchart of the included women is shown in P < 0.001), 1.76 (P < 0.001), and 0.66 (P = 0.002), respectively, while younger age had slightly but significantly positive effects on the clinical pregnancy, live birth, and miscarriage with adjusted ORs of 0.93 (P < 0.001), 0.91 (P < 0.001) and 1.12 (P < 0.001), respectively. Anyhow, high-normal TSH levels had no adverse impact on the abovementioned outcomes  (As shown in ctively) .P = 0.129). The preterm birth rates were similar between the two groups . The low-weight birth rates were also similar between the two groups . In addition, the gestational age and birth length of single live births were similar between the low-TSH and high-normal TSH groups , as were the gestational age and length of twin live births .We further compared obstetric outcomes between the low-TSH and high-normal TSH groups. Of the 1786 women included, 471 in the low-TSH group had live births, while 385 in the high-normal TSH group had live births . No signIn this retrospective study, we investigated the association between preconception TSH and the reproductive outcomes of 1786 infertile women undergoing the first fresh D3 embryo transfer at a university-affiliated hospital and found that a high-normal preconception TSH level (between 2.5–4.2 mIU/L) was not associated with lower odds of clinical pregnancy and live birth or higher odds of miscarriage. Neonatal outcomes were also comparable between low-TSH and high-normal TSH groups.Chen et al.  investigOur previous results were in agreement with other studies, which demonstrated that mildly elevated preconception TSH levels before treatment had no adverse impact on the clinical outcomes in infertile women undergoing intrauterine insemination (IUI) treatment . In addiSimilarly, Reh et al. used different TSH cutoffs in a large retrospective cohort study which included first-cycle IVF patients, and no differences in the rates of clinical pregnancy, delivery, or miscarriage were observed after adjustments were made for age . AghahosAccording to the molecular basis of TSH and thyroid hormone action during implantation and early development, thyroid hormone might be involved in endometrium preparation for pregnancy and initial trophoblast development, and thyroid hormones are essential players in the mechanisms regulating implantation and early fetal development . As a maIn contrast, Diana et al. claimed a detrimental effect of high-normal preconception TSH levels on the clinical outcomes of infertile women undergoing their first IVF cycle . DiscrepThere were also some limitations in this study. First, this was a retrospective study with data collected from a single center, and selection bias was inevitable. Second, we did not report the status of thyroid antibodies in infertile women, which might have a role in infertility and miscarriage. However, on the one hand, the existing data are controversial regarding whether thyroid antibodies are associated with infertility or adverse reproductive outcomes , 14, 20.We included infertile women as homogeneous as possible to minimize the bias that may lower the power of the impact of TSH on reproductive outcomes. We found that a high-normal TSH level did not significantly influence the reproductive outcomes of infertile women undergoing the first fresh D3 embryo transfer. Further studies are needed to test whether the results might be applicable to a wider population.


Ochlerotatus canadensis. We identified JCV in 0.6% of mosquito pools, a similar prevalence to neighboring Connecticut. We assembled the first Massachusetts JCV genome directly from a mosquito sample, finding high identity to JCV isolates collected over a 60-year period. Further studies are needed to reconcile the low vector prevalence and low rate of viral evolutionary change with the increasing number of reported cases.Jamestown Canyon virus (JCV) is a neuroinvasive arbovirus that is found throughout North America and increasingly recognized as a public health concern. From 2004 to 2012, an average of 1.7 confirmed cases were reported annually in the United States, whereas from 2013 to 2018 this figure increased over seventeen-fold to 29.2 cases per year. The rising number of reported human infections highlights the need for better understanding of the clinical manifestations and epidemiology of JCV. Here, we describe nine patients diagnosed with neuroinvasive JCV infection in Massachusetts from 2013, the year of the first reported case in the state, to 2017. Because current diagnostic testing relies on serology, which is complicated by cross-reactivity with related orthobunyaviruses and can be negative in immunosuppressed patients, we developed and evaluated an RT-qPCR assay for detection of JCV RNA. We tested this on the available archived serum from two patients, but did not detect viral RNA. JCV is transmitted by multiple mosquito species and its primary vector in Massachusetts is unknown, so we additionally applied the RT-qPCR assay and confirmatory RNA sequencing to assess JCV prevalence in a vector candidate,  Orthobunyavirus that can cause acute febrile illness, severe meningitis, and encephalitis [et al., in preparation) and other arboviruses [Jamestown Canyon virus (JCV) is an arbovirus in the genus phalitis . An RNA phalitis ; it becaphalitis ,3. Howevphalitis . Approxiphalitis ,11, and phalitis . Mortaliphalitis  and may phalitis , howeverphalitis . To charphalitis . Serologoviruses . To asseAedes intrudens and Ochlerotatus abserratus mosquitoes [Ochlerotatus canadensis in 40 of 91 locations and 9 of 10 years; JCV isolations were also made from this species with equal distribution across the state despite varying land use, suggesting that it is a primary local vector [O. canadensis pools containing 13,779 adult mosquitoes collected in Massachusetts from 2012 to 2016.The primary vector of JCV in Massachusetts is unknown, with only three isolations in the west of the state reported in the literature, from squitoes . JCV hassquitoes , with vesquitoes . A studyl vector . To addret al., in preparation). The work was approved by the Institutional Review Board (IRB) under MDPH protocol 965856 and Partners protocol 2018P001282 .Cases of JCV infection in Massachusetts between 2013 and 2017 were identified by the Massachusetts Department of Public Health (MDPH): this yielded nine cases from seven hospitals, and search of clinical records identified no additional cases. Clinical, laboratory, and imaging data were extracted from medical records. All patients were diagnosed by JCV-specific capture ELISA and PRNT using standard testing algorithms at the CDC. Archived serum was available for RT-qPCR testing from Patients 2 and 4, while acute serum and cerebrospinal fluid (CSF) from Patient 7 was tested by metagenomic sequencing as part of a separate study  from 2012 to 2016 by the MDPH and collaborating mosquito control programs. Traps were set on average every 2 days, often in multiple localities, and traps were collected the following day. Mosquitoes were stored at 4°C, taxonomically sorted, and pooled (≤50 per pool). O. canadensis pools (N = 359) were homogenized in BA-1 medium, and total nucleic acid was extracted from 140 µl of homogenate using the MagNA Pure 96 DNA and Viral Nucleic Acid Small Volume kit . An aliquot was confirmed negative for West Nile virus (WNV) and Eastern equine encephalitis virus (EEE) by RT–PCR [® VR-712™, lot #12507) using MagNA Pure Total Nucleic Acid I . To verify non-viability, extracted material was inoculated onto Vero cells grown at 37°C in BA-1 medium (Supplementary Data), subpassaged once and monitored for cytopathogenic effects for 14 days.Mosquito trapping with CDC light and COy RT–PCR . As a po® Green RNA-to-CT™ 1-Step kit , with 500 nM primers and 3 µl template in a 10 µl reaction. Conditions were: 48°C for 30 min, 95°C for 10 min, 45 cycles of 95°C for 15 s and 60°C for 30 s, and melting with 95°C for 15 s, 55°C for 15 s, 95°C for 15 s.JCV has a segmented (-)ssRNA genome: the S  segment encodes the nucleocapsid; M  encodes the surface glycoprotein; and L  encodes the RNA-dependent RNA polymerase. S is the most sampled genome segment, with 109 published sequences at the time of writing (compared to under 10 each from M and L). We designed JCV-specific PCR primers within S by aligning published JCV sequences with the closely related Inkoo virus (IV) from Northern Europe, the synonymous Jerry Slough virus (JSV) and South River virus (SRV), and 6 more distantly related orthobunyaviruses using ClustalW (Supplementary Data). The primers target a 120 bp region: F: 5′-GCAGGGTTTGTGGCATTTATGG-3′, R: 5′-TTTCCGCTCCGGTTTACGAG-3′. DNA standards were constructed spanning the target . RT-qPCR was carried out using the Power SYBRThe RT-qPCR assay was validated on the positive control RNA, and products were analyzed for size and primer dimerization by TapeStation. RNA isolated from study samples – including archived serum samples from Patients 2 and 4 and all 359 mosquito pools – were screened in triplicate alongside standards, positive control RNA, and no template controls (NTCs). Because SYBR Green dye binds dsDNA nonspecifically, nonspecific amplification can produce false positive signals at higher PCR cycles. This necessitates the use of stringent negative controls. For this assay, out of 75 NTC wells, nonspecific amplification was detected in 2 (2.7%), with Ct values of 33.9 and 37.2. Therefore, for a sample to be called positive, we required amplification in all three wells, with an average Ct of <33.9. However, some mosquito samples close to this threshold were selected for further screening by RNA sequencing in order to ensure we did not miss true positive samples with low levels of viral RNA.® III  was followed by Nextera XT  fragmentation and 18-cycle PCR amplification. Libraries were quantified  and pooled at equimolar concentration. Sequencing was performed with MiSeq or HiSeq instruments  to obtain 150 bp or 100 bp paired-end reads. If multiple JCV reads were identified in a sample, an independent library was constructed from cDNA and sequenced for confirmation.Eleven of 359 mosquito pools underwent RNA sequencing: one that was JCV RT-qPCR positive, nine with negative RT-qPCR results close to the threshold (low-level amplification in ≥1 well), and one negative control pool. Samples were treated with TURBO™ DNase  and cleaned using Agencourt RNAClean XP beads . Unbiased cDNA synthesis with random hexamer primers and SuperScriptDe novo assembly of filtered reads was attempted, followed by reference-assisted improvement. For samples with too few JCV reads to assemble contigs, reads were BLAST queried to confirm their identity. To assess the possibility of contamination between samples, reads underwent manual comparison to positive samples in this study, the positive control strain, and public JCV sequences.Reads were processed using viral-ngs   as an outgroup (Supplementary Data). Sequences were de-duplicated using CD-HIT , alignedNine cases of JCV infection diagnosed by serology were identified in Massachusetts between 2013 and 2017. Their clinical presentations, detailed below and summarized in Supplementary Table 1, shared several notable features. Most patients were generally healthy, or only mildly immunocompromised, and had a high degree of outdoor exposure. Although no patients died, nearly all had a prolonged course including fatigue, headache, altered mental status, and other neurological symptoms.Patient 1: A 57-year-old man presented with rash, myalgia, fatigue, and chills in July 2013. IgM Western blot was positive for Borrelia burgdorferi, and he was treated with doxycycline. Two days later, he developed impaired orientation, tangential thought with confabulation, and visual hallucinations. He had general myoclonus, bilateral asterixis and cogwheeling, and pharyngeal weakness from multiple cranial neuropathies. Brain MRI, obtained after lumbar puncture (LP), showed diffuse dural enhancement and chronic subdural collections. Analysis of CSF revealed lymphocytic pleocytosis with elevated protein (Babesia microti (PCR negative), and was treated for babesiosis and presumed Lyme meningoencephalitis. Ultimately, he was diagnosed with JCV, with positive IgM ELISA and a PRNT result of 1:160  was also positive by ELISA, but with PRNT 1:160 and less epidemiological risk of exposure, this was considered to represent cross-reactivity. The patient’s symptoms resolved by ten weeks after onset.Patient 4: A 63-year-old man presented to clinic in September 2014 with three days of fever, chills, and malaise. He had nausea, diarrhea, and a diffuse headache. He was febrile to 101.6 F, with no focal neurological deficits. He reported spending extensive time outdoors in both northeastern Massachusetts and southern Maine. He did not undergo brain imaging or LP. He was treated empirically with doxycycline but Lyme ELISA was negative. Serum JCV IgM ELISA was positive with a PRNT of 1:320. The patient’s symptoms completely resolved within two months.Patient 5: A 36-year-old man presented with three weeks of headache and three days of altered sensory perception in June 2015. He was afebrile, with pressured speech but no focal neurological findings. MRI showed gyriform enhancement with FLAIR hyperintensity and restricted diffusion in the right parietal cortex  and bicuspid aortic valve with ascending aortic aneurysm, recently repaired. He lived in central Massachusetts on a farm and spent substantial time outdoors. An engorged Ixodes scapularis tick was discovered during his hospitalization, and he was treated for presumed Lyme meningoencephalitis. A serum sample sent on admission was positive by JCV IgM ELISA with PRNT of 1:2560. The patient slowly improved, with persistent headache that was attributed to migraine. Two months after admission, he had continued improvement in memory and cognition and was working but had ongoing fatigue.Patient 8: A 28-year-old man presented with two days of headache and confusion in August 2017. He had no fever or abnormalities on examination. Head CT was normal, and LP was not done. The patient had persistent dull headache, neck stiffness, and fatigue 12 days later, and was evaluated in an infectious disease clinic. MRI of the brain was normal. LP, performed two and a half weeks after symptom onset, was unremarkable. The patient was otherwise healthy. He spent significant time outdoors, noted mosquito bites, and had recently travelled to Maine. Serum was positive for JCV IgM ELISA with a PRNT of 1:160. Arbovirus testing was also positive for Saint Louis encephalitis virus (SLEV) IgM with PRNT ≥1:10, however this finding was not replicated on confirmatory testing at the CDC, so likely represents a false-positive. At six-month follow-up, the patient continued to have intermittent dull headaches.Patient 9: A 40-year-old man with recent history of back pain experienced a sudden loss of consciousness in September 2017. He was afebrile, disoriented to time and place, and reported severe left frontal and eye pain. A seizure was considered likely. LP and MRI were both unremarkable. Serum was positive for JCV IgM ELISA with a PRNT of 1:160. No specific mosquito exposure was reported. His hospital stay was significant for persistent retrograde amnesia. At one month post-discharge, his memory was gradually returning, and he reportedly felt well but had not returned to work.et al., in preparation). Although based upon a limited number of samples, these results do not support routine testing for JCV RNA in immunocompetent patients, similar to other neuroinvasive arboviruses such as West Nile virus [Many of the patients described here experienced a delay in diagnosis, including empiric treatment for more common infections, before arbovirus serology was requested and results returned, highlighting the need for rapid clinical diagnostic tests. We therefore tested the feasibility of molecular detection of JCV RNA using the few residual clinical samples available. From Patient 2, an archived serum sample from approximately 1 month after symptom onset was available, and was negative by JCV RT-qPCR. From Patient 4, an archived serum sample from approximately 3 days after symptom onset was available, and was also negative by RT-qPCR. Patient 7 had been enrolled in a study of pathogen identification by metagenomic sequencing; no JCV reads were identified in CSF obtained approximately two days after symptom onset, or in plasma obtained approximately one week after symptom onset . One pool (Pool 10) was positive, with a concentration of 273 copies/µl (mean Ct 25.7). The other 358 pools were negative for JCV by RT-qPCR.Pools of RNA sequencing was performed for the positive pool, along with nine other pools whose RT-qPCR results were negative but close to the Ct threshold in ≥1 well , as wellO. canadensis mosquitoes trapped in Bristol County, Massachusetts, in June 2016. Phylogenetic analysis of the JCV S segment recapitulated the A, B1, and B2 clades described from mosquitoes in Connecticut [JCV reads from Pool 10 were successfully assembled into the complete coding sequences of the S, M, and L genome segments with mean coverage depths of 122X, 64X, and 33X respectively (accessions MN135989-91). This genome, hereafter referred to as JCV/16/MA/01, was assembled from a pool of necticut  . While these were residual samples that may have been degraded by freeze/thaw cycles, our findings are consistent with prior results; JCV has never been isolated from human samples, and one study demonstrated no detectable viremia in experimentally infected rhesus macaques, despite seroconversion [et al., in preparation).Our cases highlight several challenges associated with JCV diagnostics. Clinically, JCV is indistinguishable from other neuroinvasive arboviruses and can resemble more common infections such as Lyme meningitis, so confirming the diagnosis relies upon laboratory testing. Diagnostic testing for JCV is currently performed by the CDC’s Arboviral Diseases Branch and a limited number of state public health laboratories. Serology remains the gold standard; although PCR is faster than serology, it is unreliable for many arboviruses due to short and/or limited viremia. In our assessment of molecular tests for JCV, we did not detect JCV RNA by RT-qPCR from serum samples from two patients, one of which was obtained only three days after symptom onset. Similarly, we did not detect JCV by RNA sequencing from acute serum and CSF from a third patient (Piantadosi nversion . In genenversion  or detecnversion , Oropoucnversion , and Cacnversion . OverallIllustrating an important challenge of serology-based testing, three patients in our series had positive IgM for LACV. Like JCV, this is a California serogroup Orthobunyavirus, but is not found in New England. Prior studies have noted LACV cross-reactivity in up to 23% of patients with JCV , and it O. canadensis is important given its predominance in Connecticut [O. canadensis pools in Connecticut from 1997 to 2006 [O. canadensis may not be the primary vector for JCV. Unlike most arboviruses, JCV does not have a strict vector range [The primary vector for JCV in Massachusetts is unknown; we hypothesized that necticut . Among 3 to 2006 , suggestor range ,40, and or range ,41–43. Nor range .The full coding sequence of JCV/16/MA/01 was successfully assembled from a pool of mosquitoes trapped in Bristol County, Massachusetts, in June 2016, the first JCV genome from the state. This JCV genome is also the first sequenced directly from the primary mosquito sample, and was therefore unaffected by passaging in cell culture. JCV/16/MA/01 belongs to the rarely isolated B2 lineage . Interes


Pseudis tocantins). This genome is 15.56 kb, excluding the control region, and is similar in gene content to other hylid mitogenomes. Maximum likelihood analysis, using the mitogenomes of several anurans, indicated P. tocantins as closely related to other hylid species.In this work, we present for the first time the mitochondrial genome of a paradoxical frog ( Pseudis tocantins Caramaschi and Cruz P. tocantins has highly heteromorphic sex chromosomes , using a standard phenol:chloroform extraction protocol  occurring at high frequencies among genomic reads using RepARK script  and sequenced on an Illumina HiSeq X Ten . We detected le Edgar  and usedle Edgar , under tP. tocantins (GenBank accession number MH571152) spanned 15.56 kb, with a GC content of 43.16%. This genome is similar to those of other hylids in size, gene content, and arrangement, with an average nucleotide similarity of 70% when compared to six species from Hyla + Dryophytes. There are 13 protein-coding genes, 22 tRNA genes, and 2 rRNA genes. These genes are coded in the heavy strand, except for eight tRNA genes  and one protein-coding gene (ND6).The assembled mitogenome of P. tocantins as the sister taxon of all extant hylid species analyzed in this paper (Pseudis as a member of the Hylidae family (Faivovich et al. The resulting phylogenetic tree placed is paper . This re


Several previous studies reported a greater prevalence of dry eye syndrome (DES) among patients with psychiatric diseases. The aim of this study is to investigate the prevalence and risk factors of DES in patients with psychiatric disorders (PD) using nationwide population-based data in Taiwan.This population-based cohort study retrospectively identified patients with PD from 1997 to 2011. Patients with both PD and DES served as the DES cohort, and PD patients without DES comprised the non-DES cohort. PD was defined as a diagnosis of PD (ICD-9-CM 290–319) made by psychiatrists only, with at least three consecutive outpatient visits or at least one inpatient visit. DES was defined as a diagnosis of DES (ICD-9-CM 375.15) and a prescription for an eye lubricant . The main outcome measures were the prevalence of DES in these patients and associated risk factors.A total of 75,650 patients with PD  were included in the final analysis. The majority of patients in the DES group were women (72.6%), compared the non-DES group (57.8%). The mean age of patients in the DES cohort was 62.2 ± 14.9, which was significantly older than those in the non-DES group (50.9 ± 17.5). The patients with DES had a significantly greater likelihood of having dementia, bipolar disorder, depression, and neurotic disorders. Conditional regression analyses revealed that patients with dry eye disease were more likely to have schizophrenia (OR = 1.34), bipolar disorder (OR = 1.9), depression (OR = 1.54), and neurotic disorders (OR = 1.62). In addition, patients with DES were more likely to use 1st generation anti-psychotics (OR = 1.28) and had a lower risk of using 2nd generation anti-psychotics (OR = 0.64).The study demonstrated that among PD patients, DES is highly prevalence in certain subtypes of PD, such as depression, bipolar disorder, and neurotic disorders, after adjusting for the comorbidities. Dry eye syndrome (DES) is a common ocular surface disease worldwide. It is a multifactorial disease of tears and ocular surface DEWS 2007), causing irritation, blurred vision, burning, and foreign body sensation which affect patients’ work, daily activities, quality of life, and emotions , 2.007, causSeveral previous epidemiological studies reported a greater prevalence of DES among patients with psychiatric diseases, such as depression, anxiety, and bipolar disorder –7. HowevThe data were obtained from Taiwan’s National Health Insurance Research Database (NHIRD). The Taiwan’s National Health Insurance (NHI) program was established on March 1, 1995, and currently provides health care coverage for 99% of the country’s population, approximately 23 million people. In this study, we used the Longitudinal Health Insurance Database 2010 (LHID2010), a subset of the NHIRD, comprising 1,000,000 randomly selected NHI beneficiaries. Patients’ demographic data including gender, date of birth, income level, and healthcare data including diagnostic codes, drug prescriptions, and medical procedures contained in the database and are made available to researchers. To protect patients’ privacy, all personally identifiable information is encrypted prior to release. The NHI records diagnoses according to the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM).The study was approved by the Institutional Review Board of Taichung Veterans General Hospital, Taichung, Taiwan (CE13152B-6). In this study, the requirement for informed consent was waived because the patients’ original identification numbers were anonymized by encryption.This population-based cohort study retrospectively identified patients with PD (ICD-9-CM 290–319) from 1997 to 2011 from the LHID2010. We defined PD diagnoses as those made by psychiatrists only, with at least three consecutive outpatient visits or at least one inpatient visit. Patients with both PD and DES (ICD-9-CM 375.15) served as the DES cohort, and PD patients without DES comprised the non-DES cohort. DES was defined as a diagnosis of DES and a prescription for an eye lubricant . Subjects who had a diagnosis of DES prior to the diagnosis of PD were excluded from this study. In addition, we excluded patients younger than 18 years and any PD diagnosis related to children or infants . Moreover, patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and Sjogren’s syndrome (SS) patients were also excluded.The main outcome measures were the prevalence of DES in PD patients and associated risk factors.p-value < 0.05 in 2-tailed tests was defined as significant.The statistical analyses were conducted using the SAS 9.3 statistical package . We used Pearson’s Chi-square tests to compare the prevalence rates of different PD among patients with and without DES. In addition, the odds ratio (OR) and 95% confidence interval (CI) for gender, age, and each subtype of the PD were conducted to calculate the risk among patients with and without DES. A n = 2771), were younger than 18 years (n = 1195), had a PD diagnosis related to children or infants , or had a diagnosis of RA (n = 260), SLE (n = 105), or SS (n = 132). The remaining 75,650 patients with PD  were included in the final analysis.Among the 1,000,000 subjects in the database, 90,318 patients were identified as being diagnosed with a PD , bipolar disorder (OR = 1.9), depression (OR = 1.54), and neurotic disorders (OR = 1.62). In addition, patients with DES were more likely to use 1st generation anti-psychotics (OR = 1.28) and were less likely to use 2nd generation anti-psychotics (OR = 0.64).n = 3665/75650).The aim of the current study was to use a nationwide population-based database to evaluate the prevalence of DES in patients with PD and to investigate the associated risk factors for the period 1997 to 2011. We found the prevalence of DES in adult patients with PD was 4.84% . Dibajnia et al. showed significant decreased tear break up time in patients with bipolar disorder treated with either lithium carbonate or sodium valproate [5. DibajnOlder age has been shown to be associated with increased risk of both DES and dementia . PreviouA major strength of this study was the use of a large cohort study to investigate the association between DES and PD using data from the National Health Insurance Research Database (NHIRD). This database covers approximately 99% of the country’s residents. However, there were several limitations in this study. First, it was a retrospective study and detailed information are not available in the NHIRD, such as socioeconomic status, severity of disease, and laboratory results. Second, we used strict exclusion criteria and definitions for the diagnosis of DES. Only patients with both ICD-9-CM diagnosis code 375.15 and an anatomical therapeutic chemical code for ophthalmological lubricants (ATC code: S01XA) were included in the analysis. This selection bias might have resulted in an underestimation of the prevalence of DES in psychiatric patients, because patients with only mild dry eye symptoms may not need to use any medication. In addition, some patients may seek other medications that are not available on the NHI system.This nationwide population-based cohort study demonstrated that among PD patients, DES is highly prevalence in certain subtypes of PD, such as depression, bipolar disorder, and neurotic disorders, after adjusting for the comorbidities.


There has been research about the prevalence and risk factors of eczema, hay fever, and asthma in children, but little is known about these conditions in adults in China.To explore the prevalence of adult eczema/atopic dermatitis (AD) and its risk factors in northern China.Dermatophagoides pteronyssinu (Dp) allergen. The questionnaire was designed by specialists and included questions on the prevalence of eczema, hay fever, and asthma, socioeconomic risk factors, family history of atopy and environmental exposures. The prevalence of eczema with asthma and/or hay fever (EAH) was applied as a proxy of AD in this study.A cluster sampling randomized population-based survey was conducted using a face-to-face questionnaire combined with skin prick tests of ten common aeroallergens including nine pollen allergens and P < 0.05, logistic regression). The sensitization rate was higher in EAH compared with eczema , with weed pollen sensitization being the most common. Subjects with two or more concomitant allergic diseases had increased risk of eczema and EAH (P < 0.001). Allergen sensitization increased the risk of eczema and EAH .Overall, 2096 subjects were enrolled and completed the study. The prevalence of eczema was 15.7% (95% CI 14.3–17.4), while the prevalence of hay fever and asthma were 20.6% (95% CI 18.9–22.4) and 6.5% (95% CI 5.5–7.6), respectively. In particular, the prevalence of EAH was 5.1% (95% CI 4.4–7.0). The prevalence of eczema and EAH was significantly associated with younger age, atopy family history, high education level, urbanization, and antibiotic overuse (Adult eczema and EAH are prevalent in northern China under high pollen exposure. Socioeconomic and environmental factors affected the prevalence of adult AD in China. Dp had a particular impact on the prevalence of eczema/AD in the grassland region. Atopic dermatitis (AD) is a chronic, inflammatory cutaneous disorder that typically begins in early childhood, with a prevalence of 7–10% . AD is aTraditionally, AD is considered a disease of childhood that remits in adolescence and adulthood , 6. AlthThe diagnosis of adult AD is challenging , 20. TheThe prevalence of childhood AD is reported to be 2.5–12.94% in China –25. To oA multistage, clustered, stratified survey was carried out in the northern grassland of China from May to August of 2015 . Data regarding allergic disorders  and concomitant chronic diseases, family history of atopy, and environmental risk factors were also collected. The questionnaire was double-checked and re-answered if the quality was unsatisfactory. All participants signed informed consent. For those < 18 years, informed consent was signed by their parents or guardians. Approval to conduct this study was granted by the institutional review boards of Beijing Shijitan Hospital, the affiliated hospital of the Beijing Capital Medical University (2015 No. 3).Eczema was defined as a positive response to the following questions: “During the past year, have you had dermatitis, eczema, or any red, inflamed skin rashes?” or “Have you been diagnosed with eczema or dermatitis by a dermatologist during past year?” or typical eczema rashes were found on the study participants.Hay fever was defined as a positive response to the following questions: “During the past year, have you been diagnosed with hay fever by a doctor or health professional?” or “Did you have at least two of the four common symptoms  for at least one hour on pollen days during the past year?”.Asthma was defined as a positive response to the following question: “Have you been diagnosed with asthma by a physician during the past year?”.Subjects who were reported to have eczema combined with asthma and/or hay fever were defined as having EAH as published , 16, 21.Artemisia (Ar); Betula (Be), Chenopodium (Ch); Humulus scandens (Hu); Salix (Sa); Zea mays (Ze); Juniperus (Ju); Ulmus pumila (UI); Populus (Po), and Dermatophagoides pteronyssinus (Dp). Standardized allergen extracts  were applied. Antihistamines and other related medications were restricted at least three days prior to SPT. SPT was performed by experienced nurses following a standard protocol [All participants received skin prick tests (SPTs) of the ten common aeroallergens in the northern grassland region, which were determined by a combination of pollen monitoring and our preliminary study . The tenprotocol . The dro2 defined as lean or healthy, BMI between 25 and 29 kg/m2 defined as overweight, and BMI ≥ 30 kg/m2 defined as obese. Environmental risk factors: current smoking status , pet keeping, heating mode , overuse of antibiotics (> 3 times/year), and outdoor activity .The covariates in this study were recorded from the questionnaire. Socioeconomic factors: gender; age group; education status ; family income; and BMI, with BMI < 25 kg/mt test or Wilcoxon rank-sum test for continuous variables, and chi-square or Fisher exact test was used for categorical variables. The Bonferroni correction was performed to correct P values of multiple pairwise comparisons. Multivariate logistic regression analysis was performed to explore the risk factors related to adult eczema and estimate their odds ratios. All tests were two-sided with a significance level of 0.05.Data were processed and analyzed with IBM SPSS Statistics for Windows Version 23.0  by a professional biostatistician. Categorical data ware described as numbers and percentages. Continuous data are shown as mean ± standard deviation (SD). Differences between groups in subject characteristics were tested using a A total of 2096 adults in six regions of the northern grassland of Mongolia were enrolled and completed the study. There were 850 (40.5%) male and 1246 (59.5%) female subjects in the study cohort. Overall, 330 subjects were found to have eczema, with a prevalence of 15.7%  and EAH  was higher in urbanized areas compared with rural areas . Prevalence of positive family history of atopy was higher in EAH group than in eczema group . There was a significant difference in the prevalence of eczema among the six study areas (P = 0.007), but EAH did not vary significantly (P = 0.32)  and higher family income  were associated with increased prevalence of eczema and EAH. Environmental factors are evaluated in Table P = 0.029); however, this variation was not seen in eczema (P = 0.073). Antibiotic overuse significantly increased the prevalence of EAH , but did not increase the prevalence of eczema (P = 0.235).Socioeconomic factors that were associated with higher prevalence of adult eczema and EAH are shown in Table P < 0.001 in medium compared with low level) while medium and high education levels increased the risk of EAH  as shown by a multivariate logistic regression model. Family history of atopy increased eczema risk 1.39-fold, and increased EAH risk 2.10-fold . Antibiotic overuse increased the risk of EAH, but not eczema  . Regarding EAH, the aeroallergen sensitization rate was 53.8%, which was higher than the eczema population (P < 0.001). The sensitization rates to 10 aeroallergens are shown in Fig. P < 0.001). The sensitization of Dp was positively correlated with increasing eczema  and EAH   and EAH . Subjects with two or more combined allergic diseases were at increased risk of having eczema and EAH (P < 0.001).Allergic disease comorbidity could increase the risk of eczema and EAH Table . In partEczema represents a group of conditions with no standard clinical definition. The diagnosis of adult AD can be challenging since this terminology has been defined recently and a lack of familiarity and confusion in the definition of the condition persists –8. As inAdult AD has distinct clinical characteristics as compared to child-onset AD . Herein,2, heavy traffic and exposure to diesel exhaust could damage the skin barrier, increase trans-epidermal water loss, and increase the production of IgE [The variation of adult AD prevalence suggests a role for socioeconomic and environmental factors. No gender difference was found in the present study in contrast to other reports that noted women displayed the study-defined symptomology and clinical presentations , 29. Pean of IgE –36. Envin of IgE –36. Highn of IgE , 37–39. n of IgE . In thisn of IgE . A lowern of IgE , 41. In Another key topic addressed in this study was the comorbidities of adult AD. A higher prevalence of other allergic diseases, such as asthma and hay fever, was found in the eczema/AD group compared with the non-eczema group in our study. Thus, AD may play a role in the persistence of asthma or hay fever , 3, 9. ROne strength of this study was the evaluation of allergic sensitization for adult AD. It is intriguing that the prevalence of AD is highly affected by aeroallergen sensitization. High pollen concentration, as we reported , may conWe performed a population-based investigation and analyzed the allergen sensitization of eczema/AD. Efforts to minimize selection bias were made. However, some limitations should be mentioned. First, eczema was defined as self-reported cases of doctor-diagnosed “eczema” or “atopic dermatitis”. This might overestimate the prevalence of adult eczema due to possible confusion with irritant or contact dermatitis. Second, the prevalence of adult AD may include both childhood persistent AD or adult-onset AD . In thisThe prevalence of adult eczema and AD in the northern grassland area of China was 15.7% and 5.1%, respectively. Adult eczema/AD was positively associated with increased asthma and hay fever. Significant predictors of adult eczema/AD included age, place of residence, higher education level, higher family income, family history, heating mode, overuse of antibiotics, allergen sensitization, and comorbid atopic diseases.