Membrane distillation (MD) has shown promise for concentrating a wide variety of brines, but the knowledge is limited on how different brines impact salt scaling, flux decline, and subsequent wetting. Furthermore, past studies have lacked critical details and analysis to enable a physical understanding, including the length of experiments, the inclusion of salt kinetics, impact of antiscalants, and variability between feed-water types. To address this gap, we examined the system performance, water recovery, scale formation, and saturation index of a lab-scale vacuum membrane distillation (VMD) in long-running test runs approaching 200 h. The tests provided a comparison of a variety of relevant feed solutions, including a synthetic seawater reverse osmosis brine with a salinity of 8.0 g/L, tap water, and NaCl, and included an antiscalant. Saturation modeling indicated that calcite and aragonite were the main foulants contributing to permeate flux reduction. The longer operation times than typical studies revealed several insights. First, scaling could reduce permeate flux dramatically, seen here as 49% for the synthetic brine, when reaching a high recovery ratio of 91%. Second, salt crystallization on the membrane surface could have a long-delayed but subsequently significant impact, as the permeate flux experienced a precipitous decline only after 72 h of continuous operation. Several scaling-resistant impacts were observed as well. Although use of an antiscalant did not reduce the decrease in flux, it extended membrane operational time before surface foulants caused membrane wetting. Additionally, numerous calcium, magnesium, and carbonate salts, as well as silica, reached very high saturation indices (>1). Despite this, scaling without wetting was often observed, and scaling was consistently reversible and easily washed. Under heavy scaling conditions, many areas lacked deposits, which enabled continued operation; existing MD performance models lack this effect by assuming uniform layers. This work implies that longer times are needed for MD fouling experiments, and provides further scaling-resistant evidence for MD. The most commonly used process for water desalination is reverse osmosis (RO), primarily due to its comparatively low energy consumption ,2. HowevOn the other hand, membrane distillation utilizes thermal energy to provide the driving pressure force, which does not deteriorate significantly for high salinity feed ,13,14,15Numerous studies have analyzed the feasibility of RO-MD processes, and several have shown that pure water and salt deposits can be obtained from concentrated feeds, with water recovery in the range of 20–60% ,23,24,25In fact, several studies investigated the long-term operation of membrane distillation technology ,28,29,30The objective of this work was to determine the practicability of concentrating a variety of water types in a lab-scale VMD apparatus to achieve a high water-recovery ratio in long-term performance tests. Additionally, we investigated the limitations of the process due to scaling. In phase I of this investigation, the membrane was characterized using low-salinity feeds (Linz tap water) and an aqueous sodium chloride solution. In phase II, water recovery ratio and flux reduction due to scale formation of a synthetic RO brine were compared to the results for the low-salinity feeds. Finally, in phase III, the effect of using an antiscalant on permeate flux was analyzed.2. The laboratory apparatus consisted of a feed container, a stirred membrane cell, a distillate collection vessel , a magnetic stirrer , an external condenser , a platform balance , and a vacuum pump , as illustrated in The experiments were performed with a lab-scale VMD apparatus  using a f) and membrane water recovery. The concentration factor was calculated as , and SPK is the solubility product of CaCO3 in molar units. We found similar solubility trends in the speciation analysis of feed water shown in The alkaline scaling mechanism depends primarily on the temperature, the concentration of the brine, and the hydraulic conditions in the test cell . Additioy a role . The soly a role . The fol3 nucleation was around 72 h, and it decreased as supersaturation increased. Using the synthetic RO brine as a feed for the MD did therefore not result in a fast reduction in permeate flux. However, salt deposition on the membrane surface occurred after 82% recovery, which resulted in a gradual reduction in the permeate flux. Note that temperature and concentration polarization phenomena can facilitate precipitation of deposits on the membrane [As illustrated in membrane ,45,46,472 (resulting from bicarbonate in the water) passing through membrane pores together with the water vapor. One might suspect that the complete wetting of large membrane pores may have occurred, causing an increase in electrical conductivity. However, as shown in The higher initial permeate conductivity may be due to CO3), calcite, and dolomite (CaMg(CO3)2). Among these, dolomite had the highest saturation index in both the synthetic RO brine feed without antiscalant and its corresponding retentate solution. However, studies have shown that temperature has a stronger influence on dolomite precipitation than the saturating index [A multi-component aqueous speciation equilibrium model is performed to calculate the distribution of aqueous species and saturation indexes based on water elements concentrations provided in ng index . Similarng index , supersaThe effect of antiscalant on system performance was also studied to investigate the possibility of achieving a higher recovery ratio and of extending the time until scaling and the beginning of crystallization on the membrane surface.−2 h−1 to 1.59 kg m−2 h−1 after 125 h. The initial permeate electrical conductivity was around 15 µS/cm within the first 10 h of the experiment, then decreased to 2.50 µS/cm, and gradually increased to 3.70 at the end of the experiment. The brine electrical conductivity increased linearly from 9.47 mS/cm to 47 mS/cm, while a final recovery ratio of 88% was achieved. Compared to the system without antiscalant, the initial permeate flux was higher, while the permeate electrical conductivity did not increase to more than 15 µS/cm. Further, the antiscalant influenced brine electrical conductivity, and kept it below 47.5 mS/cm, while the recovery ratio remained the same  than the untreated system (86.60 mS/cm). Furthermore, using antiscalant increased the time until membrane scaling from 28 h to 59 h. Scaling – and consequently wetting—occurred when the amount of salt  in the system reached approximately 680.0 g/L water.3 and CaSO4) [As previously discussed, deposition on the membrane surface caused a reduction in permeate flux, contributed to more wetting, decreased membrane performance, and reduced heat efficiency . Scalingd CaSO4) . However−2 h−1 and 2.0 to 44.8 µS/cm for various feed types. The highest permeate flux of 3.98 kg m−2 h−1 with a permeate electrical conductivity of 2 µs/cm was achieved for the system with low-feed salinity (Linz tap water) at a feed temperature of 75 °C and a gauge pressure of −30 mbar. The long-term tests led to average flux reductions by 49% and 50%, corresponding to water recovery ratios of 91% and 88% after 128 h and 148 h of operation for the synthetic brine feed systems with and without antiscalant, respectively.The long-running lab-scale VMD experiments for water desalination were performed using a flat sheet hydrophobic (PTFE/PP) membrane to investigate the highest possible water recovery and to examine the effect of scaling on system performance for various feed waters. A series of preliminary experiments were designed to test the behavior of the membrane with a low-salinity feed (Linz tap water) and aqueous sodium chloride solution (5.25 g/L). The study’s performance emphasized the importance of longer duration experiments and demonstrated scaling resistance of MD.Importantly, our results showed that combining RO and VMD increases water recovery from 40% to 91%, which corresponds to a brine concentration factor of up to 11.1. Note that the application of the synthetic RO brine as feed for VMD did not result in an immediate decrease in permeate flux compared to using low-salinity feed. Rather, permeate flux started to decrease at a water recovery rate of 82% due to deposition of salts on the membrane surface. In addition to high retentate concentrations, we observed scaling in VMD; however, large areas of the membrane remained without visible scaling. Scaling on the membrane surface was caused by the decomposition of bicarbonates present in the brine under feed temperature conditions. Substantial membrane scaling actually reduced the concentration of salts in the retentate. Notably, in all cases, surface scaling could be washed away effectively by cleaning the membrane surface with distilled water.Furthermore, salt deposits around membrane pores were found to alter membrane hydrophobicity, which led to membrane wetting. The use of Genesys LF antiscalant delayed both flux reduction and membrane wetting. However, it also led to a decrease in the average permeate flux.There were numerous observations that are inconsistently considered in models of scale, along with their impact on performance. These included scale deposits being patchy, ease of cleaning, ability to scale without wetting in some conditions, and the strong delayed time-dependent behavior of scaling.Future investigations will consider the effect of stirring on system performance and scaling development and its morphology. Fluid circulations within the membrane module affect the permeate flux significantly. Specifically, high fluid circulation reduces ion residence time on the membrane surface and thus surface crystallization. Similar findings have been reported by other researchers ,69.Overall, the experiments show that VMD has the potential to be integrated with RO to achieve higher water recovery ratios.


More than 11 million people suffer from coronary heart disease (CHD) angina in China, showing high morbidity and mortality rates. Yufengingxin (YFNX) is a commonly used Chinese patent medicine in CHD angina treatment. The purpose of this protocol is to evaluate the effectiveness and safety of YFNX for the treatment of CHD angina.A systematic search of randomized controlled trials related to the effectiveness and safety of YFNX in the treatment of CHD angina will be performed from relevant databases, including PubMed, Cochrane Library, EMBASE, Chinese National Knowledge Infrastructure (CNKI), Wanfang Database, Chinese Scientific Journal Database (VIP) and Chinese Biomedical Literature Database (CBM). We will screen all the literatures from the database inception to November 1, 2020. The data including study ID, study characteristics, methodological information, patients information, interventions, comparisons and outcomes will be extracted. The frequency and duration of angina attacks will be served as the primary outcome. Review Manager 5.3 and STATA 14.0 software will be used for data analysis.This systematic review will provide strong evidence for the effectiveness and safety of YFNX in the treatment of CHD angina.INPLASY2020110040. Since the end of the last century, cardiovascular disease has become a leading cause of death among Chinese residents. At present, 11396,000 people in China suffer from CHD. The mortality rate of urban/rural residents reaches 115.32 per 100,000 and 122.04 per 100,000, respectively, and it shows an increasing trend year by year. Every year, nearly 910,000 patients receive percutaneous coronary intervention therapy. CHD seriously affects peoples quality of life and puts a huge economic burden on society.Coronary heart disease (CHD) angina, a common cardiovascular disease, is caused by coronary artery stenosis or spasm, which leads to myocardial ischemia and hypoxia. Therefore, to explore novel strategies for alleviating symptoms of angina, especially those with no obvious side effects, becomes an urgent task. Recently, a high-quality systematic review concluded that traditional Chinese medicine in the treatment of CHD might be effective in alleviating angina symptoms, abnormal myocardial perfusion and neurological deficits, and could be used as a complementary therapy for the treatment of CHD.Nitrates are the most commonly used drugs for relieving angina pectoris. However, the use of nitrates has been found to have some side effects in clinical practice, such as dizziness, blushing, nausea, and loss of appetite. High-performance liquid chromatography analysis showed that the main active ingredient of YFNX was puerarin, which should not be less than 13 mg/tablet as stipulated by the Pharmacopoeia.,9 Puerarin  is a kind of isoflavone compound, which has a strong activity in the cardiovascular system. It can dilate the coronary blood vessels, reduce the oxygen consumption of the myocardium, inhibit the inflammatory response, and protect the cardiomyocytes.–12Yufengningxin (YFNX), which is extracted and processed from Radix Puerariae, is a commonly used Chinese patent medicine for the treatment of CHD. It has the functions of relieving spasm and relieving pain, enhancing brain and coronary blood flow, and improving angina symptoms. Because of its remarkable effects, it was included in the Pharmacopoeia of the People's Republic of China in the 2020 edition.,14 At the same time, YFNX can also reduce the risk factors of CHD, including lowering blood pressure and low-density lipoprotein cholesterol.,16 In addition, animal experiments have further confirmed that YFNX has a protective effect on myocardial ischemia induced by isoproterenol hydrochloride in mice, improving the hypoxia tolerance of mice and prolonging the average survival time of mice with myocardial hypoxia. However, there is still no strong evidence to indicate the effectiveness and safety of YFNX in treating CHD angina. Therefore, a high quality systematic review and meta-analysis of the effectiveness and safety of YFNX in treating CHD angina will be carried out to provide a reference for clinical use.Some studies have confirmed that YFNX can significantly reduce the frequency and duration of angina in patients with CHD, and improve clinical efficacy.2 The current protocol was registered with INPLASY (registration number: INPLASY2020110040). If there are any adjustments in this study, we intend to make revisions and updates in the final publication.This review will be performed and reported in strict accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) Statement.2.12.1.1We will include all relevant randomized controlled trials (RCTs) regarding the application of YFNX to the treatment of CHD angina, with no limitation of the language and the type of publication.2.1.2By following the World Health Organization (WHO) diagnostic criteria for CHD, 2 kinds of CHD angina will be included, including stable angina pectoris (SAP) and unstable angina (UA). We will not consider the patients gender, age, ethnicity and region.2.1.3The control group accepted conventional therapy, such as aspirin, clopidogrel, β-blockers, statins, calcium channel blockers, renin-angiotensin-aldosterone system blockers, nitrates, etc. And the intervention group took YFNX combined with conventional therapy. The various dosage forms of YFNX will be included, such as tablets, capsules, dripping pills, granules and so on. Its dosage and duration will not be limited. The study in which other Chinese medicines were used will be excluded.2.1.42.1.4.1The primary outcomes will be the frequency and duration of angina attacks.2.1.4.2The secondary outcomes will include clinical efficacy rate, the efficacy rate in an electrocardiogram, scores of TCM syndrome, blood pressure, low-density lipoprotein cholesterol, fibrinogen, haematocrit and adverse events.2.22.2.1We will perform a systematic search of the relevant databases, including PubMed, Cochrane Library, EMBASE, Chinese National Knowledge Infrastructure (CNKI), Wanfang Database, Chinese Scientific Journal Database (VIP) and Chinese Biomedical Literature Database (CBM), from their inception to November 1, 2020. The English search terms include YFNX, CHD, angina pectoris and RCTs. The detailed search strategy for PubMed is shown in Table 2.2.2We will also retrieve grey documents and clinical trial registers, such as WHO International Clinical Trials Registry Platform (ICTRP), the Chinese Clinical Trial Register (Chi CTR) and the Clinical Trials, to supplement the electronic databases.2.32.3.1We will import all studies into Endnote X9 software. Two researchers (Chao Liu and Jing Bai) will independently extract the potentially eligible studies by screening the titles and abstracts in accordance with the inclusion criteria. After removing irrelevant and duplicate studies, they will further scan the full text to assess their eligibility. All studies selected by the researchers will be cross-checked, and if there are any disagreements, they will be discussed and solved by a third author (Jie Wang). The literature screening and selection processes will be performed in accordance with the PRISMA flow chart shown in Figure 2.3.2Two researchers (Chao Liu and Jing Bai) will independently extract and manage the data from the finally included studies according to the pre-defined data collection form. The form will include study ID, study characteristics , methodological information , patients information , interventions and comparisons , and outcomes (primary outcomes and secondary outcomes). If there is any unclear or missing information, the author of the original study will be consulted. Any disagreements will be solved through discussion.2.3.3The Cochrane “risk of bias” tool will be used to evaluate the risk of bias in each included study by 2 independent authors. The main items include selection bias , performance bias, detection bias, attrition bias, reporting bias and other biases. According to the actual situation of the research, each bias is divided into 3 levels, including low risk, high risk, and unclear. If there are any disagreements in the assessment, we will reach a consensus by discussing and negotiating with a third author (Lanchun Liu).2.3.4We will use the mean difference (MD) and 95% confidence interval (CI) to analyze the continuous data. For the dichotomous data, the rate ratio (RR) and the 95% CI will be used to expressed the results.2.3.5As for the missing or incomplete information in the included studies, we will contact the author to obtain the missing or incomplete information by email or other means. If the information is not accessible, the impact of the missing data should be discussed, and we will only analyze the available data.2.3.6I2 test will be used to evaluate the statistical heterogeneity. If I2 ≤ 50%, that means there is no significant heterogeneity among studies and a fixed-effects model should be used to calculate the effect size; otherwise, it means there exists significant heterogeneity.The 2.3.7I2 ≤ 50%) exists between the studies, the fixed-effects model will be used for meta-analysis. Conversely, if evident heterogeneity is found, a random-effects model will be applied to evaluate the outcome data, and a subgroup, sensitivity analysis or descriptive analysis will be conducted to analyze the potential reasons for heterogeneity. If quantitative synthesis is not appropriate, we will perform a descriptive analysis.The RevMan 5.3 software  will be used for data synthesis. If small heterogeneity (2.3.8If there are more than 10 trials in the study, the funnel plot analysis will be performed to evaluate publication bias. In the case of fewer than 10 trials, we will conduct a quantitative analysis through the Egger test by using the STATA 14.0 software.2.3.9If there have obvious heterogeneity and more than 10 trials among the included studies, a subgroup analysis based on the type of angina pectoris, the dosage form of YFNX and adverse events should be conducted.2.3.10To ensure the robustness of analytical conclusions, we need to perform a sensitivity analysis by eliminating each study individually or changing the category of the effect model.2.3.11 will be used to classify the quality of evidence in the systematic review into 4 levels: “high quality”, “moderate quality”, “low quality”, and “extremely low quality”.The GRADE tool2.3.12This article collects published articles for data analysis and does not involve the individual privacy of patients, so ethical approval is not necessary. The results of this review will be presented at relevant conferences and published in peer-reviewed journals.3 and autophagy via the Akt signaling pathway, increasing superoxide dismutase, decreasing creatine kinase and methylene dioxyamphetamine. Therefore, we designed the systematic evaluation protocol by using the latest data to test the effectiveness and safety of YFNX in the treatment of CHD angina. It is hoped that this study could find more rigorous medical evidence for the application of YFNX to the treatment of CHD angina, thus providing a reference for clinical practice. However, there are still some potential limitations in this study. First, there still lacks of a high quality, multi-center and large sample clinical trial, which will affect the authenticity of the evidence. Second, different dosage forms and doses of YFNX may lead to significant heterogeneity of results. Therefore, more rigorous, large-sample, high-quality RCTs should be carried out in future studies to confirm the clinical efficacy of YFNX.CHD angina affects physical function and the quality of life, and imposes a broad burden on both patients and caregivers. YFNX has significant clinical effects, and its main active ingredient puerarin can protect against myocardial ischemia/reperfusion injury by inhibiting inflammationConceptualization: Chao Liu.Data curation: Jing Bai, Lanchun Liu.Formal analysis: Jing Bai, Jialiang Gao.Funding acquisition: Jie Wang.Methodology: Chao Liu, Lanchun Liu.Project administration: Jie Wang.Resources: Jing Bai.Software: Lanchun Liu, Jialiang Gao.Writing – original draft: Chao Liu, Lanchun Liu.Writing – review & editing: Chao Liu, Jing Bai.


The prevalence of prediabetes and diabetes is increasing rapidly, and 5% to 10% of prediabetic patients will develop diabetes every year. Diabetes causes major health problems as well as a large economic burden. Human studies have demonstrated the beneficial effects of probiotics, prebiotics, and synbiotics supplementation in prediabetes. However, there are no systematic reviews that explore the therapeutic efficacy of probiotics, prebiotics, and synbiotics supplementation in patients with prediabetes. Therefore, we aim to synthesize the existing evidence evaluating the effectiveness and safety of probiotics, prebiotics, and synbiotics supplementation in prediabetic patients.We will search PubMed, EMBASE, Cochrane Library, Web of Science, the Clinical Trials.gov website, China National Knowledge Infrastructure, China Science and Technology Journal Database, and Wanfang Data Knowledge Service Platform from inception to August 2020. Additionally, the search will be conducted in multiple languages. Search terms are keywords and medical subject headings related to prediabetes, probiotics, prebiotics, and synbiotics. The primary outcomes are differences in glycated hemoglobin and fasting blood glucose. The secondary outcomes are differences in fasting insulin, homeostasis model assessment of insulin resistance, quantitative insulin sensitivity check index, and adverse events. The meta-analysis will be performed using the Revman5.3.0 software provided by the Cochrane Collaboration.Our study will systematically evaluate the effectiveness and safety of probiotics, prebiotics, and synbiotics supplementation in prediabetes.The findings of this study will provide the best available evidence for probiotics, prebiotics, and synbiotics in the treatment of prediabetes, and provide a strong basis for clinical treatment. The number of diabetic patients worldwide is projected to rise from 382 million in 2013 to 592 million in 2035. It is estimated that 470 million people worldwide will suffer from prediabetes by 2030, and 5% to 10% of prediabetic patients will develop diabetes every year. Diabetes causes major health problems as well as a large economic burden. The direct annual cost of diabetes worldwide exceeds US$827 million. Moreover, diabetics lose a great deal of labor value due to reduced productivity and production time. Consequently, timely and effective preventive measures in prediabetes are a reasonable way to prevent the diabetes epidemic and lessen the healthcare cost.Prediabetes, typically defined as glycaemic parameters above normal levels but below diabetes thresholds, is a metabolic state between normal blood glucose and diabetes, including impaired fasting glucose, impaired glucose tolerance, and impaired fasting glucose combined with impaired glucose tolerance. Effective interventions conclude lifestyle changes and medications.–7 However, in clinical practice, the adherence to lifestyle changes in prediabetic populations is low, and the compliance of using hypoglycemic drugs is relatively poor. Thus, to find a nonhypoglycemic agent is of great significance in preventing the conversion of prediabetes to diabetes. Recent studies indicate that alterations in gut microbiota play a major role in the pathogenesis of diabetes.–10 It is reported that when the intestinal dysbacteriosis, the production of lipopolysaccharide increases, the release of pro-inflammatory cytokines increases, and a series of nonspecific inflammatory responses occur in the body, which interferes with insulin signal transduction, induce insulin resistance and lead to hyperinsulinemia, as well as trigger chronic low-level inflammatory responses of islet cells and metabolic endotoxemia, and ultimately, lead to destruction and apoptosis of islet β cells.,13 Microecological preparations, including probiotics, prebiotics, and synbiotics, have beneficial health effects on the host when administered in sufficient amounts. Studies suggest that certain microecological preparations can exert anti-diabetic effects in different studies,,15 improving glycemia, insulin sensitivity and inflammatory markers in subjects with type 2 diabetes.–19 Several human studies have also demonstrated the beneficial effects of microecological preparation supplementation in prediabetes.,21One study has shown that early intervention in prediabetes can reduce the risk of developing type 2 diabetes by 58%.To the best of our knowledge, up to date, there are no systematic reviews that explore the therapeutic efficacy of probiotics, prebiotics, and synbiotics supplementation in patients with prediabetes. The aim of this systemic review and meta-analysis is to synthesize the existing evidence evaluating the effectiveness and safety of probiotics, prebiotics, and synbiotics supplementation in prediabetic patients.22.12.1.1Only randomized controlled trials (RCTs).2.1.2All adult patients who met the accepted diagnostic criteria for prediabetes will be included in this study without discrimination of race or gender.2.1.3Only RCTs comparing probiotics, prebiotics, and synbiotics supplementation with placebo will be included in this study. Also, the minimum duration of treatment is 8 weeks.2.1.4RCTs with placebo treatment for at least 8 weeks will be included as the controls in this study.2.1.52.1.5.1(1)Differences in glycated hemoglobin;(2)Differences in fasting blood glucose.2.1.5.2(1)Differences in fasting insulin;(2)Differences in homeostasis model assessment of insulin resistance;(3)Differences in quantitative insulin sensitivity check index;(4)Incidence and severity of adverse events.2.2We will utilize the computer to retrieve the following databases: PubMed, EMBASE, Cochrane Library, Web of Science, the Clinical Trials.gov website, China National Knowledge Infrastructure, China Science and Technology Journal Database, and Wanfang Database from inception to August 2020. Additionally, the search will be conducted in multiple languages. Search terms are keywords and medical subject headings related to prediabetes, probiotics, prebiotics, and synbiotics. Keywords and medical subject headings used in this study include: “prediabetic state,” “prediabetes,” “glucose intolerance,” “impaired glucose tolerance,” “impaired fasting glucose,” “probiotics,” “prebiotics,” “synbiotics,” “randomized controlled trials,” and “clinical trial.” The search strategy will be determined by 2 researchers after several pre-searches. The PubMed search strategy is outlined in Table 2.3 Furthermore, the meta-analysis protocol of this study was created according to the PRISMA-P guidelines. The process of literature identification and screening is shown in Figure The literature included in the meta-analysis will be independently screened by 2 reviewers according to the eligibility criteria listed in this study. In the process of literature selection, the title and abstract will be read first, and the full text will be further read to determine the final inclusion after excluding the obvious irrelevant literature. The reporting quality of the included studies will be independently assessed by 2 reviewers using the PRISMA evaluation scale.2.4According to the pre-designed data extraction table, data will be extracted independently by 2 reviewers. All studies that meet the eligibility criteria will be extracted as follows:(1)Basic information, including study design, author, country, publication year source of literature, sample size, and grouping;(2)Basic characteristics, including age, gender, and the number of participants;(3)Methodological description, including randomized method, blind method, allocation concealment, losses to follow-up, and assessed risk of bias;(4)Interventions and controls characteristics, including time and type of interventions and controls;(5)Study outcomes, including primary and secondary outcome indicators, as well as adverse events.During the data extraction process, the extracted data will be checked by a third reviewer for verification. Moreover, if there is any disagreement in the process of data extraction, it needs to be re-examined and discussed, and even the judgment of a third reviewer.2.5We will give a call or sent an e-mail to the author to obtain relevant data. The study will be deleted if the missing data is unavailable. Besides, we will perform a sensitivity analysis of the missing data and evaluate its impact on this study.2.6The quality of the study will be assessed by 2 reviewers in accordance with the “biased risk assessment” Handbook 5.3 provided by the Cochrane Collaboration. There are 7 main items:(1)Random allocation method;(2)The allocation scheme is hidden;(3)Blind methods are used for subjects and treatment planners;(4)Blind method for measurement results;(5)The integrity of the resulting data;(6)Selectively report the results of the study;(7)Other sources of bias.Finally, the “low risk of bias,” “high risk of bias,” and “uncertain risk of bias” of the study will be judged. If the opinions are inconsistent, they will be resolved through group discussion.2.7I2 statistical value. When there is statistical homogeneity between the studies (I2 ≤ 50% or P > .10), a fixed effect model will be employed to assess the difference. Conversely, when there is statistical heterogeneity between the studies (I2 ≥ 50% or P < .10), the evaluator will conservatively assess the difference using a random effects model. Besides, the results of individual studies will be summarized descriptively if fewer studies are unable to carry out a meta-analysis.The meta-analysis will be performed using the Revman5.3.0 software provided by the Cochrane Collaboration. The relative risk and the 95% confidence intervals will be used for the dichotomous variables. And the mean difference or standard mean difference will be used as a therapeutic statistic for the continuous variables, along with 95% confidence intervals. The heterogeneity test utilizes the 2.82.8.1A funnel plot analysis will be performed on the indicators with the largest number of literatures (≥10 articles) to detect the possibility of publication bias.2.8.2We will perform a subgroup analysis of the different dosage and duration of interventions, respectively.2.8.3To ensure the stability of the results, we will remove each study in turn for sensitivity analysis to assess whether the deleted study affects the overall effect.2.8.4After completing the data extraction and evaluation analysis, we will use the GRADE pro 3.2 software to assess the overall quality of the evidence and the strength of the recommendation.3The increasing number of research on probiotics, prebiotics, and synbiotics for prediabetes leads us to believe that these interventions have improved glycemia and insulin sensitivity. The purpose of this systemic review and meta-analysis is to objectively estimate the effects of probiotics, prebiotics, and synbiotics supplementation on glycemic control and insulin metabolism in prediabetic patients. To interpret the results accurately, we will ascertain the influence of efficacy and safety in different dosages of probiotics, prebiotics, and synbiotics and different duration of treatment. The strength of this systematic review is that screening, data extraction, and quality assessment will be performed independently by 2 reviewers. Herein, this systematic review and meta-analysis will be the first one, which employ rigorous methods to identify and collate the best available evidence assessing the effectiveness and safety of probiotics, prebiotics, and synbiotics supplementation in prediabetes, therefore informing clinical treatment decisions.Conceptualization: Xuqin Du, Chunguang Xie, Lipeng Shi.Data curation: Xuqin Du.Funding acquisition: Qionghui Liu.Investigation: Xuqin Du.Methodology: Lipeng Shi, Xuqin Du.Project administration: Lipeng Shi, Xuqin Du.Software: Lipeng Shi.Supervision: Chan Yang.Validation: Hong Gao.Writing – original draft: Lipeng Shi, Xuqin Du.Writing – review and editing: Lipeng Shi, Xuqin Du.Lipeng Shi orcid: 0000-0002-4478-0622.


In this study, nickel phosphorus (NiP)–titanium (Ti) nanocomposite coatings (NiP-Ti nanoparticles (TNPs)), containing various concentrations of Ti nanoparticles (TNPs) were deposited on high strength low alloy (HSLA) steel through electroless deposition processing. The concentrations of 0.25, 0.50 and 1.0 g/L TNPs were dispersed in the electroless bath, to obtain NiP-TNPs nanocomposite coatings comprising different Ti contents. Further, the effect of TNPs on the structural, mechanical, corrosion, and heat treatment performance of NiP coatings was thoroughly studied to illustrate the role of TNPs into the NiP matrix. Field emission scanning electron microscope (FESEM) and energy dispersive spectroscopy (EDX) results confirm the successful incorporation of TNPs into the NiP matrix. A substantial improvement in the mechanical response of the NiP matrix was noticed with an increasing amount of TNPs, which reached to its ultimate values  at NiP-0.5TNPs coatings composition. Likewise, the electrochemical impedance spectroscopy measurements confirmed a tremendous increase in the corrosion inhibition efficiency of the NiP coatings with an increasing amount of TNPs, reaching ~96.4% at a composition of NiP-0.5TNPs. In addition, the NiP-TNPs nanocomposite coatings also unveiled better performance after heat treatment than NiP coatings, due to the presence of TNPs into the NiP matrix and the formation of more stable (heat resistant) phases, such as Ni Nickel phosphorus (NiP) coatings have propitious mechanical  and anticorrosion properties, so they have applications in industries like automotive, oil and gas, electronics, and aerospace ,2,3. The3N4, TiN, BN, Al2O3, TiO2, SiO2, SiC, B4C [2O3 [2 [2 and Si3N4 can increase the corrosion resistance properties of NiP coatings [Further, to enhance the properties of electroless NiP coatings, incorporation of metallic or ceramic (micron or nano) particles into the NiP matrix has been carried out, leading to the development of composite coatings. For example, the inclusion of CSiC, B4C ,12,13,14SiC, B4C ,16,17 caB4C [2O3  and TiO2 [2O3 [2  into thecoatings ,19. Mostcoatings ,21. In ccoatings . Therefocoatings ,24,25,26coatings . Likewiscoatings ,29. TherIn the present study, NiP-TNPs composite coatings were developed by reinforcing the NiP matrix with various concentrations  of Ti nanoparticles (TNPs) through the electroless deposition process. HSLA steel was chosen as a substrate to target its application for the aerospace industry. Further, the effect of TNPs concentration on structural, mechanical, corrosion, and heat treatment behavior of NiP coatings was examined to clarify the superior performance of these nanocomposite coating systems. The coating morphology and chemical composition were studied using field emission scanning electron microscope (FESEM), EDX, and X-ray diffraction (XRD) techniques. The mechanical evaluation of the developed coatings was performed through the microhardness and nanoindentation techniques. The anticorrosion response of the developed coatings was assessed through the electrochemical impedance spectroscopy (EIS) and potentiodynamic polarization techniques in 3.5 wt. % NaCl solutions.3, with an elemental composition, as shown in Electroless deposition of NiP and NiP-TNPs composite coatings was carried out on a high strength low alloy (HSLA) steel substrate, which was obtained from Wugang Chang Yu Long Industry and Trade Co., Ltd., ShenZhen, China. The substrate samples have dimensions of 30 × 25 × 2 mm2SO4) as the Ni source, and sodium hypophosphite (NaPO2H2) as the reducing agent and as a P source. The substrates were ground with different grit SiC abrasive papers  successively, to achieve flat and smooth surfaces followed by soap water and ethanol cleaning. The steel substrate samples were washed with deionized water and activated with a 15% solution of H2SO4 for 30 s. After surface activation, the pretreated specimens were again rinsed with distilled water and immersed directly into the deposition (electroless solution) bath. The preparation of the electroless plating bath was performed by mixing specific quantities of Nichem A and Nichem B, as provided by the manufacturer, in DI water to obtain 1 L of the electroless solution. The temperature of the bath was kept at 86 ± 2 °C for 90 min, which is the deposition time, and the deposition bath was agitated with a magnetic stirrer at 300 rpm to avoid settling of TNPs during deposition processing. For each nanocomposite coating composition preparation, a specific amount of TNPs was added into 1 L of the electroless bath, and the solution was mixed with the ultrasonic probe for 0.5 h, before initiating the deposition process to eliminate any chances of their agglomeration and settling of TNPs during deposition. Then, the cleaned specimens were immersed in the electroless bath, and the deposition process started under the same previous conditions and lasted for 90 min. After that, the coupons were removed, rinsed, and dried. This procedure was repeated for each concentration of TNPs to obtain the different nanocomposite coatings. For simplification, the developed NiP-TNPs nanocomposite coatings were designated as per the TNPs concentrations, which is dispersed into the electroless bath. In this perspective, the obtained coatings samples were labeled as NiP, NiP-0.25TNPs, NiP-0.5TNPs, and NiP-1.0TNPs, respectively. For a clear understanding of electroless deposition processing of NiP-TNPs nanocomposite coatings, a schematic diagram is shown in An analytical grade Titanium powder with 99.9% purity , comprising an average particle size of 70.0 nm, was used as reinforcement material (nanoparticles). Different concentrations 0.25, 0.50, and 1.0 g/L of TNPs were used in electroless deposition (electrolyte) bath for preparation of NiP-TNPs coatings. The electroless deposition solution used in this study was a standard commercial-grade (Nichem 2500 (A and B) from Atotech Inc., Berlin, Germany) solution, with nickel sulfate  technique, using a PAN analytical X’pert Pro Cu (Ka), with a scanning rate of 0.02° in the 2θ range from 20° to 80°. Surface morphology and elemental analysis were performed using FESEM (field emission scanning electron microscope) from Nano-450, Netherland, also contain EDX (energy dispersive spectroscopy) analyzer. The topographic study was performed using AFM (atomic force microscope) from Oxford Instruments, Goleta, CA, USA. A coating thickness gage meter (PosiTector 6000) from DeFelsko, Proctor Avenue Ogdensburg, NY, USA was used to measure the thickness of coating samples. The mechanical response of the coatings was studied through microhardness and nanoindentation techniques. Microhardness measurements were performed with Vickers microhardness tester from FM-ARS9000, USA, and nanoindentation measurements were performed using MFP-3D nanoindenter head attached with AFM. For the corrosion study in 3.5 wt. % NaCl solution, electrochemical impedance spectroscopy (EIS) and potentiodynamic polarization (PP) methods were adopted. All electrochemical measurements were performed in an electrochemical cell, having three electrodes systems attached with a GAMRY 3000 potentiostat from Warminster, PA, USA. The calomel electrode was used as a reference, the graphite rod was used as a counter, and coating samples were used as a working electrode. An area of 1 cmThe heat treatment of the developed coatings was commenced in the electric furnace . The selected coatings samples were heat-treated at 400 °C for different time intervals  in the open air. Initially, the weight of each sample was measured, followed by heating. After heating at 400 °C, for each time interval, the heat-treated samples were weighed again, and the obtained weights were recorded. For weight measurements, the analytical grade balance  with 0.0001 g sensitivity, was utilized. The coated samples subjected to the high-temperature treatment were analyzed using FESEM, AFM, and XRD techniques to study the surface morphology, topography, and composition of the formed phases.Further, to have more details on the surface characteristics of the developed coatings, AFM surface analysis was carried out. The 3D surface profile-images of NiP and NiP-TNPs coatings are shown in EDX test was performed to perceive the co-deposition of TNPs into the NiP matrix and to study their effect on the chemical composition of the NiP coatings. The phases investigation of the developed coatings was performed through XRD. Prior to perform mechanical testing (microhardness and nanoindentation), the thickness of the coating samples was measured using thickness gage meter. Coating thickness measured values for all compositions are presented in 50) value contributing to ~28% increase in the microhardness when compared to NiP coatings. The increase in the hardness values can be regarded as the effect of dispersion hardening and grain refinement strengthening evolved by the incorporation of TNPs in the NiP matrix. In the dispersion hardening process, it can be assumed that the TNPs can act as an obstacle that does not deform during the deformation, such that the moving dislocations have to bypass the obstacles by changing their deformation path, whereas the grain refinement process can also strengthen the materials by providing obstacles to the dislocation motion at grain boundaries. In our case, TNPs dispersed into the deposition bath may provide various heterogeneous nucleation sites leading to accelerating the nucleation rate, which reduces the lateral growth of the grains and thus ultimately forms a fine structure. This argument can be visualized in FESEM images presented in The microhardness test results of NiP, NiP-0.25TNPs, NiP-0.5TNPs, and NiP-1.0TNPs nanocomposite coatings are presented in Further, to evaluate the mechanical characteristics of NiP and NiP-TNPs nanocomposites coatings, nanoindentation was performed by using the nanoindenter head (Berkovich diamond indenter tip) connected to AFM, using a maximum 1 mN indentation force at loading and unloading rate of 200 μN/s and having dwell time of 5 s. The load-displacement curves of NiP and NiP-TNPs composite coatings having various concentrations of TNPs are shown in The electrochemical impedance spectroscopy (EIS), and potentiodynamic polarization (PP) corrosion measurements were carried out in 3.5 wt. % NaCl solutions at room temperature, to illustrate the corrosion performance of the electroless NiP-TNPs coatings with different Ti nanoparticles concentrations.Rs, Rpo, and Rct are solution, pore, and charge transfer resistances, respectively, CPEcoat donates the constants-phase element for the coating and CPEdl is the constant-phase element for the double layer. The low–frequency time constant (CPEdl and Rct) demonstrates to the substrate/coating interface, whereas the combination of CPEcoat and Rpo represents the high–frequency time constant that corresponds to the coating/solution interface. The W is referred to the Warburg diffusion resistance, which clarifies the existence of electrolyte diffusion. The CPE is a pseudo-capacitive element, which is used instead of the regular one due to the surface roughness and the coating inhomogeneity, which is assigned to the irregular thickness of the deposited coatings, in addition to the non-uniform current distribution at the surface [n is the CPE exponent. The deviation from ideal behavior is expressed by the exponent n (0 ≤ n ≤ 1). The ideal resistor is indicated when n equals zero, whereas the ideal capacitor is acted when n equals 1 [The bode-phase angle plots and the corresponding Nyquist-plots, which are measured at OCP, for the developed coatings are shown in  surface . The imp surface ;(1)ZCPEequals 1 ,45.2. The impedance value has increased by one order magnitude for electroless NiP coating compared to that of the substrate. The formation of the hypophosphite layer causes the increased impedance value of the NiP coating due to the reaction of the phosphorus with water [Rct is observed at NiP-0.5TNPs (18.6 KΩ·cm2), with approximately 96.5% protection efficiency, which is reduced to 15.4 KΩ·cm2 at NiP-1.0TNPs, with approximately 95.6% protection efficiency, as shown in Rct value is due to the increase in the content of TNPs, which may generate defects, causing more inhomogeneity in coatings. As in th water ,47. Thisth water , which lth water . It can Moreover, it is noticed that the phase-angle plots of the NiP coatings in the absence and presence of TNPs have the same shape and position, as shown in Nyquist plots for HSLA steel, NiP, and NiP-TNPs composite coatings with different content of TNPs are depicted in Rpo) of the NiP-TNPs coatings. The values of Rpo are given in Rpo) of NiP-TNPs coatings increases with the addition of TNPs and reaches its highest value at NiP-0.5TNPs composition. Furthermore, the decrease in the amount of constant phase element for NiP-TNPs nanocomposites coatings as compared to HSLA steel and NiP coating proves their better corrosion-resistant behavior. It is worth mentioning that the inhibition efficiency (I.E) of the coatings was calculated by using the following equation [Rct2 and Rct1 are the charge-transfer resistance of the substrate and coating, respectively. The inhibition efficiency for NiP and NiP-TNPs is given in Furthermore, the decrease in porosity of the NiP coating with the addition of TNPs also helps to increase the pore resistance (equation .I.E (%)icorr (corrosion-current density), Ecorr , bc and ba (anodic and cathodic Tafel slopes) that acquired using the Tafel extrapolation method, and the corrosion inhibition efficiency (IE%) for all coatings. The inhibition efficiency (IE%) for the coated samples derived from the data obtained from Tafel plots is obtained by applying the following equation [icorr1 and icorr2 are the corrosion current density of the substrate and coating samples, respectively. The corrosion-potential (Ecorr) for HSLA steel was around −764 mV, while that for the NiP coating is shifted toward more positive value (−527 mV), which is further anodically shifted to −383, −311, and −388 mV by the addition of TNPs  in the NiP matrix. The reduction in the corrosion potential of the NiP-TNPs nanocomposite coatings compared to NiP coatings proves their better corrosion resistance. The more negative value of the corrosion potential of HSLA steel indicates that the steel’s surface is more susceptible to corrosion. Moreover, a noteworthy decrease in the corrosion-current density values of the coated samples compared to the values for the substrate is observed, such that the icorr value is dropped from 69 µA·cm−2, in the case of the substrate, to 8.2 µA·cm−2 for NiP coating. A further drop in the icorr value to 6.5 µA·cm−2 takes place at NiP-0.25TNPs coating, signifying an increase in the corrosion resistance of the NiP coating in the presence of TNPs nanoparticles. As the concentration of the TNPs particles is increased into the NiP matrix, the icorr of the NiP-TNPs nanocomposite coating decreases and reaches to the highest drop at NiP-0.5TNPs composite coating composition, as shown in ba, bc) indicates the change in the mechanism of iron dissolution due to the deposition process. The results of EIS and potentiodynamic polarization measurements are consistent and confirmed with each other.equation .(IE %) For the heat treatment study, NiP, NiP-0.5TNPs nanocomposite coating samples were selected and heat-treated at 400 °C in an open furnace, for different time intervals . AFM was also performed on heat-treated samples to quantify the surface characteristics. The 3D surface topographic images of NiP and NiP-0.5TNPs, coatings are shown in 3P phases  coatings containing various content of TNPs were successfully deposited on HSLA steel through the electroless deposition technique. The FESEM and EDX analysis confirmed the progressive co-deposition of TNPs into NiP matrix, whereas phase pure structure formation was perceived by X-ray analysis. Microhardness and nanoindentation results report the significant influence of TNPs on the mechanical properties of NiP coatings that reached to its ultimate values for the NiP-0.5TNPs nanocomposite coating composition. Similarly, the EIS results confirm the excellent corrosion inhibition efficiency of the NiP coatings with an increasing amount of TNPs, reaching to ~96.4% with NiP-0.5TNPs composite coating composition. Moreover, the heat resistance (oxidizing resistance) behavior of the NiP coating was remarkably increased, due to the presence of TNPs into the NiP matrix and the formation of more stable (heat resistant) phases, such as Ni


Human exhaled breath consists of more than 3000 volatile organic compounds, many of which are relevant biomarkers for various diseases. Although gas chromatography has been the gold standard for volatile organic compound (VOC) detection in exhaled breath, recent developments in mid-infrared (MIR) laser spectroscopy have led to the promise of compact point-of-care (POC) optical instruments enabling even single breath diagnostics. In this review, we discuss the evolution of MIR sensing technologies with a special focus on photoacoustic spectroscopy, and its application in exhaled breath biomarker detection. While mid-infrared point-of-care instrumentation promises high sensitivity and inherent molecular selectivity, the lack of standardization of the various techniques has to be overcome for translating these techniques into more widespread real-time clinical use. Even in ancient times, human beings used to relate the odor of breath to diseases. Ancient Greek physicians assessed the aroma of human breath for disease diagnosis. Nebelthau mid-1800s) identified acetone in the breath of diabetes patients, while Anstie 1874) isolated ethanol from the breath of alcoholics [800s iden, and N2 ,4. More  isolated, and N2 ,6. Figur, and N2 ,44,45,46, and N2 ,13. For , and N2 . Higher , and N2 ,18,19,20Similarly, sulfur-containing compounds in breath are the result of incomplete metabolism of methionine in the transamination pathway. Moreover, nitrogen-containing compounds are at elevated levels in human breath for liver impairment or uremia. Nitric oxide has been identified as a biomarker of airway inflammation such as in asthma, allergic rhinitis, eosinophilic bronchitis, and chronic obstructive pulmonary disease (COPD) ,22,23,242, H2O, O2, and N2 (% levels) next to much lower concentrated VOCs (ppm–ppt levels), which may mask the presence of other biomarkers for certain detection techniques. Hence, analysis techniques with pronounced sensitivity and selectivity are required for identifying and quantifying disease-related biomarkers against the highly variable breath matrix background.The list of VOCs and inorganic compounds and their relation to potential disease states, as reported in various studies, is shown in Conventional breath analysis methods are usually based on offline sampling. Gas chromatography (GC) based separation coupled with mass spectrometry (MS) remains the ‘gold standard technique’ for trace gas analysis. Owing to its historical presence and analytical capabilities, GC–MS capitalizes on extensively developed libraries, which facilitate rapid compound identification at sensitivities down to ppt levels. However, these techniques involve manual sampling procedures and sample preparation ,29,30,31Alternative techniques like electronic nose sensors have found rapid developmental interest for their applications in small, affordable, point-of-care systems in breath analysis. Predominately, conducting polymer-based and metal–oxide semiconductor (MOS)-based arrays of sensors are used for determining multiple analytes. However, they are limited in molecular selectivity, high moisture sensitivity, and power demand .Alternatively, ion mobility spectrometry (IMS) techniques such as those based on time-of-flight (TOFIMS) and differential ion mobility (DMS) have been reported for the detection of VOCs. DMS involves the separation of different ion species owing to differences in their ion mobilities in low and high electric fields. High-frequency asymmetric waveform field is used for the filtering of other species while selectively allowing ions of a particular species through the filter by tuning a low amplitude compensation field. DMS has resulted in a cost-effective, compact sensor without moving parts that allows real-time gas analysis ,40,41. HDriven by the need for point-of-care (POC) medical instrumentation, laser spectroscopic-based noninvasive human exhale analysis has drawn increasing attention during the last decades since the development of new mid-infrared (MIR) laser sources ,44,45. PA basic schematic of the setup for MIR laser absorption spectroscopy is given in Quantum cascade lasers and interband cascade lasers  in the MIR range have potential application, where many molecules relevant in exhaled breath diagnostics exhibit strong rovibrational absorptions are present. QCLs offer wide tunability and pronounced output power, leading to molecular selectivity, sensitivity, and improved signal-to-noise ratio for the detection of trace biomarkers in the exhaled breath matrix. Li et al. have used the ICL-based TDLAS approach for ethane detection in an exhaled breath at 3.34 µm, which is indicative of lung cancer and asthma. A detection limit of 1.2 ppbv was achieved at 4 s data acquisition time . Ghorban2H4) using the spectral signature in the 10 µm band of the CO2 laser [2H6) detection in breath samples in the 3.34 μm region. They compared the performance of CALOS with gas chromatography with flame ionization detection (GC–FID). Repeated tests concluded that the spectroscopy setup could detect ethane in less than 1 min, making it more suitable for continuous monitoring of patients, whereas chromatography required 30 to 60 min [A variation of the CRDS technique is the CALOS, where, in contrast to the CRDS technique, a continuous wavelength (CW) source is used to tune one of the resonance modes of the cavity, allowing energy to build up inside the cavity ,58,59,60O2 laser . Later, Though such techniques offer high sensitivity and can be adopted for continuous real-time monitoring of patients, such methods need excellent tuning of the cavity length. The optical cavity or multi-pass cell is also sensitive to mechanical vibration. Even the availability of either tunable light sources or high reflectance mirrors precisely for a particular wavelength of the target gas sample limits this technique from being widely used. Moreover, the need for high reflectance mirrors makes it relatively more expensive when compared with the other optical methods.Recent advances in MIR waveguide technology have the potential to design advanced and compact instrumentation for trace gas analysis in this spectral regime. Hollow-core waveguides (HCW) in this region (3–20 µm) can facilitate the development of highly compact and sensitive trace gas sensing devices with potential usage in POC scenarios. Hollow-core photonic bandgap waveguides (HC-PBW) absorption spectroscopy for methane detection in the 3.4 µm region has been demonstrated by Nikodem et al. with sensitivities at the ppm-level . For higRecently, a novel concept of substrate-integrated hollow waveguides has been introduced ,68,69,70Alternatively, hollow core waveguides have also been used in fibre enhanced Raman spectroscopy (FERS) ,72. FERSPAS is a zero-background technique and the PA signal is less affected by scattering. Hence, it has been widely used for trace gas detection at the part per billion or even part per trillion levels. In photoacoustic spectroscopy, as the name suggests, light energy is converted into sound energy in a series of steps. The modulated or pulsed light is focused into a gas cell containing the target sample to be analyzed. If the frequency of the light source matches with the vibrational frequency of the gas molecule to be analyzed, the light is absorbed. Periodic amplitude modulation in the light source causes a periodic acoustic vibration in the gas cell. This acoustic vibration is detected by the microphone and converted into an electrical signal ,78,79,80On the other hand, broadband sources matching with broadband spectra of species of interest can be used, which allows low power based multiple-gas sensing. Recently, QEPAS based on quartz tuning forks (QTF) as a sound transducer for the PAS technique has been increasingly used for selective and sensitive sensing ,90,91,922 laser operating in the 9 and 10 µm wavelength range. They were able to detect 100 ppb ammonia-detection sensitivity [Many studies have shown that ammonia in exhaled breath can be used for detecting chronic kidney disease (CKD), because, in patients with CKD, the accumulated urea cannot be excreted by the kidneys, but is degraded by the salivary urease into ammonia, which is exhaled through the breath. A variety of medical conditions, including liver and kidney disorders ,94, as wsitivity . Lewickisitivity . On the sitivity .Lipid peroxidation that is the reaction between omega-3 fatty acids and reactive oxygen species releases ethane in the human body, which can be identified in the human breath. Ethane has also been noted as an indicator of oxidative stress, which in turn has been said to play an essential role in the pathophysiology of several common diseases. Studies show that cancer, cardiac disease neurodegenerative disease, psychiatric illness, stroke, and diabetes patients have high oxidative stress, and hence it has been proposed to have any involvement in the initiation of disease such as in cancer ,100. Mor2H4 concentration in elderly patients (age 70–80 years) [2H4 sensors were reported previously using a 3.32 µm DFB laser (1.5 mW) to achieve a minimum detection limit of 63 ppm at 25 s averaging time [2H4 sensor by exploiting the C2H4 spectra near 10.5 µm. The CW DFB QCL with ~23 mW power output at the target wavelength near 10.5 µm achieved a minimum detection limit of 50 parts per billion (ppb) at an averaging time of 70 s [Ethylene, as a biomarker, has been studied in dialysis patients. Studies show a relation between oxidative stress and fatality in patients with renal failure, especially elderly patients. A conventional PAS sensor using a CW carbon dioxide laser (10.53 µm) was developed to monitor C0 years) . The ething time . Wang et of 70 s .2 laser in the 10.6 um range for the detection of acetone. A group of healthy individuals was studied against a group of patients with type 2 diabetes mellitus (DM). In the group with type 2 DM, the acetone range was to vary between 101 and 162 ppb, while in healthy individuals, the acetone range was between 15 and 85 ppb [For patients with diabetes, the body cannot synthesize insulin to break down glucose in the blood to provide energy. Therefore, the body undergoes lipolysis, causing decarboxylation of acetoacetate, leading to the production of significantly increased concentrations of acetone in the breath ,111. Hend 85 ppb .−1 (λ ~ 5.263 μm). The minimum detection limit achievable is ~5 ppbv with a 1 s data acquisition time [Nitric oxide is an important biomarker for chronic obstructive pulmonary disease (COPD). The patients often exhibit symptoms that alter their performance statuses such as productive cough, worsening dyspnea, peripheral muscle weakness, and nutritional abnormalities. Measurement of exhaled nitric oxide (eNO) is a non-invasive method of assessing airway inflammation. In patients with COPD, the peripheral airway (bronchioles) is the leading site of obstruction and inflammation ,114, andion time . The estMethane as a breath gas is of considerable interest because it is considered as a potential biomarker for stomach inflammatory diseases and colorectal cancer. Various studies demonstrate that high lipid, low-fat diet, and elevated bile salts, as well as the presence of colonic anaerobic bacteria, are a source of methane in the intestine, which then traverse the intestinal mucosa and are absorbed into the systemic circulation. As it has low solubility, it is rapidly excreted by the lungs. Breath methane analysis has been shown in various studies for the diagnosis of carbohydrate malabsorption syndromes and small intestinal bacterial overgrowth and, if the exhaled methane is more than one ppm as compared with ambient levels, the patient is considered a methane producer. Bauer et al. presented the development of a Raman amplifier system operating at 1651 nm and its application for trace gas sensing with a miniaturized 3D printed PAS cell. The system exhibited high sensitivity towards methane sensing with the least detection limit of 17 ppb at a signal acquisition time of 130 s .4 detection in the 1.65 µm region was developed, where multiple absorption lines of methane were utilized to produce the photoacoustic signal. A bandpass filter with a full width at half maximum (FWHM) of 12 nm was chosen with a centre wavelength at 1.65 µm where the interference owing to moisture is minimal, as shown in As the amplitude of the photoacoustic signal is directly proportional to the input power and the absorption cross-section of the target gas, in recent years, many research groups have used QCL and ICL in the mid-infrared and THz regions, which are considered as the molecular fingerprint regimes. Furthermore, the QEPAS technique, along with a QTF, has been used to fabricate small, compact PAS systems . PeterseWhile the potential advantages of breath analysis are apparent, the variability of breath measurements has been quite high, and thus their potential has not been fully utilized to date. Though chromatography is considered the gold standard in breath gas analysis, real-time monitoring is not feasible via such laboratory-based techniques probing discrete samples. On the other hand, laser spectroscopic methods can be applied to real-time monitoring of breath gases. With the advancement in MIR lasers, compact handheld devices can be established with high sensitivity, molecular selectivity, and reproducibility. Even though such laser spectroscopic techniques offer sensitivities in the ppb to ppt concentration range, there are still several improvements required to render MIR spectroscopy a useful clinical tool in routine breath gas monitoring. This an issue for all sensing techniques, as MIR spectroscopies need to deal with the uncertainty on how the presence and/or up/down regulation of selected biomarkers may be mapped onto specific diseases, disease progression, or therapeutic interventions. For example, ethane can be indicative of cancer, cardiac disease neurodegenerative disease, psychiatric illness, stroke, and diabetes. Hence, the pathobiology of the breath compounds has to be extensively studied for establishing diagnostically relevant and usable correlations between biomarkers and the disease. As several studies have shown that, indeed, multiple rather than individual VOCs giving rise to pattern changes may be used to more reliably associate with a particular condition, MIR diagnostics are certainly at the forefront of multi-component sensing techniques utilizing fingerprint patters for detecting several VOCs. Appropriate selection of broadly tunable QCL, ICL, OPO, or supercontinuum light sources enables addressing such complex fingerprints, and may aid in further lowering detection limits, especially in broadband PAS measurements. In conclusion, even though more work certainly needs to be done on MIR sensing technologies as well as on establishing reliable pathobiologically relevant breath biomarker panels, appropriate MIR sensing systems may differentiate between healthy and diseased individuals in a statistically sound fashion appear to be on the horizon.


Biswamoyopterus (Rodentia: Sciuridae: Pteromyini) was once considered to contain three species, Biswamoyopterusbiswasi from northeastern India, B.laoensis from central Laos and B.gaoligongensis from southwest China, all identified from morphological characteristics of one or two specimens. However, based on similar morphological characteristics of two samples of the genus Biswamoyopterus collected recently from northern Laos and northern Myanmar, and the small genetic distances on mitochondrial DNA and nuclear DNA between them, the results strongly support these two samples as representatives of the same species. The phylogenetic analyses strongly support Biswamoyopterus as an independent genus of Pteromyini, as a sister group to Aeromys. Biswamoyopterusbiswasi is distributed in the northern Indo-China peninsula, where it is exposed to a series of threats, such as intense hunting activity, illegal trade, and rapid habitat loss; this should warrant its classification as critically endangered according to the International Union for Conservation of Nature (IUCN) Red List criteria. Here, the molecular data for genus Biswamoyopterus and two new specimen records from northern Laos and northern Myanmar are presented.The flying squirrel genus  Mammalia: Rodentia: Sciuridae: Pteromyini), occurring in northern coniferous forests to the tropical lowlands of North America and Eurasia, are great masters of gliding locomotion using well-developed membrane structures  found in northeastern India, B.laoensisB.gaoligongensisKIZ 034924), found in southwest China , has conducted several biodiversity expeditions to the northern Indo-China peninsula  issued by Biotechnology and Ecology Institute, Ministry of Science and Technology of Lao PDR, and permission (1567/XTBG/2017) issued by the Forest Research Institute, Forest Department, Ministry of Environmental Conservation and Forestry of Myanmar.All samples used in this study were obtained by the Biswamoyopterus and ten of Petaurista) were collected from northern Myanmar and northern Laos during the expedition of 2014–2018  was collected from a local market in Putao county , Kachin State, Myanmar ; detailed sequence information has been listed in Suppl. material Twelve flying squirrel samples  ; the Institute of Zoology, Chinese Academy of Sciences (IOZ) ; and the Guangdong Entomological Institute (GDEI) ; or using documented literature (According to the taxonomic assignments of Wilson and Reader 2005), pelage and skull characteristics can be discriminated using traditional methods and compared with those of other genera using specimens (Appendix I) retained in the Kunming Natural History Museum of Zoology, terature . Followi005, pelaBB Breadth of braincase,BH Braincase height,CBL Condylobasal length,DL Diastema length,FL Frontal length,GPB Greatest palatal breadth,IBG Inter bullae gap,IOB Interorbital breadth,LAB Length of auditory bulla,LBP Length of bony palate,LIF Length of the incisive foramina,MB Mastoid breadth,MH Mandible height,ML Mandible length,MRTL Mandibular tooth row length,MWN Maximum width of nasals,MYTL Maxillary tooth row length,NL Nasal length,OB Orbit breadth,ONL Occipitonasal length,PL Palate length,POB Postorbital breadth,PPL Postpalatal length,RB Rostrum breadth,WAAM Width of auditory bullae across the external auditory meati,WPFM Width of the bony palate at the first upper molar,ZB Zygomatic breadth,ZH Zygomatic height,P Premolars,M Molars.X, MX) upper premolars and upper molars, and Subscript  lower premolars and lower molars.Superscript  and mitochondrial 12S and 16S ribosomal DNA of flying squirrels were amplified using PCR with universal primers described previously . PCR mixtures contained approximately 100 ng of template DNA, 1 μL (10 pmol) of each primer, 5 μL of 10× reaction buffer, 2 μL of dNTPs (2.5 mM of each), and 2.0 U of Taq DNA polymerase, in a total volume of 50 μL. Reactions were carried out in a Veriti Thermal Cycler  and always included a negative control. Segments of the nuclear genes encoding the inter photoreceptor retinoid-binding protein (eviously . FragmenDNA sequences were edited using the DNASTAR 5.0 (DNASTAR Inc.) program and aligned using the CLUSTALW algorithm in MEGA 6.06, with default parameters . IdenticBI) in MRBAYES 3.2.6 (MCMC) analysis approximated posterior distributions with one cold and three heated chains, and samples of the trees and parameters were drawn every 100 steps from a total of one million MCMC generations; three additional runs were conducted beginning with random trees. The 50% majority rule consensus of the post-burn (using a burn-in of 25%) for all generations was computed for the four runs. Trees were visualized using FIGTREE 1.4  in a complex honeycomb pattern, the orbital regions are large and there is an incision on the edge of each orbit, the postorbital process is strong and curves down a little, the zygomatic plate is slant, the zygomatic arch is stronger with lower connection to the squamosal, the mastoid process is comparatively smaller, but the occipital condyle is strong.3 is cone-shaped and on the inside of the front of P4; overall, the crown surface of P4 appears as a triangle with three well-developed cusps on the labial side and one large cusp on the lingual side, and its labial side length is slightly longer than those of M1, M2, and M3. M1 and M2 are approximately equal in size; both have two well-developed cusps on the labial side, and one large cusp and one smaller cusp on the lingual side. There is a smaller cusp on the posterior transverse ridge of P4, M1, and M2. Compared with P4, M1, and M2, M3 is the smallest; its lingual side cusp is larger than the cusp on the labial side, and its later crown surface becomes a U-shape, with a small depression in its center.The mandible is strong, with the coronoid process developed, and the condylar process has a developed articular surface; the angular process is developed and curved towards the inside at its bottom. The upper incisors are strong and positioned vertically downwards; their outer surfaces are yellowish, without any orange. P4 to M3, the teeth enlarge gradually, and there are two labial and lingual cusps on each of them ; there is also a smaller cusp between, and slightly internal to, the two labial cusps on each of them. Different levels of depression occur in the centers of the crown surfaces of P4, M1, M2, and M3, with the largest in M3.The outer surface of the lower incisors is yellowish, the same as for the upper incisors; however, the inside part of the inner surface sinks deeply, making the outside margin sharp. From PTaxon classificationAnimaliaRodentia SciuridaeMorphological description ofsp. L3545C70733-0599-57C7-BF11-7B2D5090D7BDBiswamoyopteruslaoensis specimen (NUoL FES. MM.12.163) from central Laos. However, specimen M644 from northern Myanmar shares some key characters that have been used to distinguish the three known species from each other  from central Laos; the coloration of venter, tail, and ear tufts could pertain to either B.biswasi (specimen ZSI 20705) from northeastern India or B.gaoligongensis (specimen KIZ 034924) from southwestern China, which are very similar.The sample L35 from northern Laos shares the same pelage color of the tuft hair at the base of the ear and side of the neck , mitochondrial 12S (823 bp), and 16S (535 bp) ribosomal DNA recovered similar tree topologies. The results showed that Eupetaurus, Aeromys, and Biswamoyopterus  as a reciprocally monophyletic clade  variation was 0.51–5.47% , close to 0.13% for intraspecific variations of Pteromyscuspulverulentus.For the nuclear gene Biswamoyopterus and other genera ranged from 5.2 to 12.8% , close to 0.4% for intraspecific variations of Petauristaphilippensis, and the same as 0.6% of Pteromysvolans.For the mitochondrial 16S ribosomal DNA sequences, the range of original intergeneric variation was 2.9–14.6% Table . The genBiswamoyopterus specimens L35 from northern Laos and M644 from northern Myanmar are confirmed as representing the genus Biswamoyopterus. However, Biswamoyopterus sp. M644 shares many key characters with all three known Biswamoyopterus species. Since each Biswamoyopterus species has been described on the basis of only one or two samples, it is possible that the observed morphological differences are the result of intraspecific variation. If so, it is plausible that all known Biswamoyopterus specimens might in fact be conspecific.According to morphological comparisons of our samples and those from previous studies , BiswamoB.laoensis from B.biswasi mainly by 1) the large distance of 1250 km between the localities of the two species and 2) the different pelage colors present mostly on the ventral side: “white but washed with a faint orange-rufous” in B.biswasi versus “essentially orange” in B.laoensis. These factors were also true for samples L35 and M644, being separated by a long distance of more than 1000 km and different ventral pelage colors. Biswamoyopterus specimens M644 and L35 indicate that further studies should be performed to shed light on the relationships among B.biswasi, B.laoensis, and B.gaoligongensis.It was further implied by the molecular evidence that samples L35 and M644 belonged to the same species, with the smallest nuclear and mitochondrial DNA genetic distance among interspecific variations for any of the studied flying squirrel species (Tables Biswamoyopterus as an independent genus within Pteromyini, acting as a sister group to Aeromys (Figure Biswamoyopterus and other genera are greater than many of the intergeneric variations (Tables Biswamoyopterus is a separate flying squirrel genus distinct from every validly described genus. We note that DNA sequences for genus Aeretes cited in the literature may be based on mistaken institutional identifications, as reported recently by The molecular phylogenetic analysis strongly supported s Figure . For nucs Tables , 5. BothBiswamoyopterus were found. We therefore propose that Biswamoyopterus should be classified as critically endangered on the IUCN Red List, due to a series of threats on the Indo-China peninsula that include intense hunting, illegal trade, and rapid habitat loss (Biswamoyopterus, further studies including biodiversity expeditions covering the whole Indo-China peninsula should be performed. With respect to biogeography, members of the genus Biswamoyopterus inhabit the northern Indo-China peninsula, which belongs to one of the global biodiversity hotspot regions (Biswamoyopterus and to make every effort to conserve them.During the expedition of 2014–2018, only two samples of tat loss . In orde regions . The mec


Regarding biomarkers of iron status, plasma hemoglobin (Hb), ferritin, and transferrin saturation (%) levels were significantly lower in ID and IDA groups. The prevalence of ID and IDA were 3.7% and 2.7%, respectively, in infants under six months of age, but increased to 20.4% and 6.6%, respectively, in infants above six months of age. The healthy group had a higher total iron intake than ID and IDA groups, mainly derived from infant formula. The total dietary iron intake was positively correlated with infants’ Hb levels. Compared with formula-fed infants, the logistic regression revealed that the odds ratio for ID was 2.157  and that for IDA was 4.196 (95% CI: 1.780–9.887) among breastfed infants (p < 0.001) after adjusted for all confounding factors . In conclusion, our results determined that breastfeeding was associated with an increased the prevalence of ID and/or IDA, especially in infants above six months. This suggests that mothers who prolonged breastfeed after six months could provide high-quality iron-rich foods to reduce the prevalence of ID and IDA.Iron deficiency (ID) and iron deficiency anemia (IDA) typically occur in developing countries. Notably, ID and IDA can affect an infant’s emotion, cognition, and development. Breast milk is considered the best food for infants. However, recent studies have indicated that breastfeeding for more than six months increases the risk of ID. This study investigated the prevalence of ID and IDA, as well as the association between feeding type and iron nutritional status in northern Taiwan. A cross-sectional study was conducted on infants who returned to the well-baby clinic for routine examination from October 2012 to January 2014. Overall, 509 infants aged 1–12 months completed the iron nutritional status analysis, anthropometric measurement, and dietary intake assessment, including milk and complementary foods. The results revealed that 49 (10%) and 21 (4%) infants in their first year of life had ID and IDA, respectively, based on the World Health Organization criteria. Breastfed infants had a higher prevalence rate of ID and IDA than mixed-fed and formula-fed infants ( Breast milk is considered the optimal nutrition and healthy food for the first six months of life, and breastfeeding is typically complemented with other foods from six months of age until at least 12 months for nearly all infants . Anemia In addition to inadequate iron intake, reduced bioavailability of dietary iron, increased iron requirements, and chronic blood loss is common causes of iron deficiency (ID). ID is currently the most common micronutrient deficiencies worldwide, and a World Health Organization (WHO) survey in 2008 revealed that globally, approximately 293 million  preschool-age children and approximately 56 million  pregnant women suffer from anemia, among which approximately 50% cases are attributable to ID ,11.Recent studies have indicated that iron content in breast milk is low ,13,14, aA cross-sectional study was conducted in three hospitals, namely, Shin Kong Wu Ho-Su Memorial Hospital, Taipei Medical University Hospital, and Shuang Ho Hospital, from October 2012 to January 2014. Overall, 2804 healthy infants aged 1–12 months who came to the well-baby clinic for routine vaccination were screened for eligibility. The inclusion criteria for infants and mothers enrolled were as follows: No systemic diseases  during pregnancy, age below one year, healthy and without any disease, as determined by a pediatrician. Infants with premature birth, congenital diseases , growth disorders, diagnosis of gastrointestinal disorders , and thalassemia were excluded.The Ethics Committee of Shin Kong Wu Ho-Su Memorial Hospital and Taipei Medical University approved this study in accordance with the International Ethical Guidelines for Biomedical Research Involving Human Subjects and ethical principles of the Declaration of Helsinki (20120901R and 201308004). Written informed consent was obtained from all participants or legal representatives before the study procedures were performed.The contents of the questionnaire were validated by experts, including the infant’s basic information, such as gestational weeks, chronological age, birth weight, birth length; anthropometrics, such as body length, body weight, and head circumference; and dietary intake. Gestational weeks, chronological age, birth weight, and birth length were recorded per the medical chart. Body length and body weight were measured using an infantometer and weighing scales. The head circumference was measured by applying a plastic tape around the forehead (above the eyebrows) and the occipital protuberance. These measurements were converted to percentiles according to the growth charts for Taiwanese children released by the Ministry of Health and Welfare. The diet was assessed using a semi-quantitative frequency method, and the included food items were tofu, chicken, pork, beef, fish, egg yolk, viscera, vegetables, rice, wheat, baby rice cereal, baby wheat cereal, formula milk, juice, and puree. Complementary food and formula milk intakes were quantified based on standard bowls, spoons, and feeding bottles. Iron intake from the complementary food was calculated using the Nutritional Chamberlain Line, Nutritionist Edition, version 2002 . Iron intake from formula milk and baby cereal was calculated according to their nutrient labels. Iron content in breast milk was obtained through actual measurement. Breast milk intake was obtained from mothers’ reports. If the mothers were directly breastfeeding their infants, the total volume of breast milk intake was assessed according to the report of Lyu et al. . Pre-preg for 10 min at 4 °C and immediately sent to Central Laboratory, Shin-Kong Wu Ho-Su Memorial Hospital, for analysis.Breast milk from the mothers’ unelated breast was collected using an electric milking machine  for 20 min, which was then mixed, dispensed, and ice-cooled at −80 °C until analysis. Blood samples (4 mL) of infants were collected through an arterial puncture of one arm into vacutainers with and without anticoagulant ethylenediaminetetra-acetic acid (EDTA) by a trained technician. Serum was collected after centrifugation at 1400× Complete blood count was determined using an automatic blood cell analyzer . Ferritin was detected using a chemiluminescent immunoassay . Serum iron was analyzed using the ferrozine method . Total iron binding capacity (TIBC) is the ability of transferrin to bind with iron that was measured by chemistry analyzer using dedicated reagents . Transferrin saturation  represents the percentage of transferrin bound to iron ions, calculated by dividing serum iron concentration by TIBC and multiplying the result by 100.R value of ≥0.99, coefficient of variation (CV) at 2%, and a recovery rate of 80–120%.Aliquots of 0.5 mL of the mixed breast milk sample was added to 1.5 mL of 70% nitric acid and 0.5 mL of 30% hydrogen peroxide, separately. After mixing and allowing the samples resting for one night, the mixture was digested in a 50 mL polypropylene digestion bottle at 95 °C for 1 h. After cooling at room temperature, the digested sample was diluted using 50 mL of deionized water. Subsequently, 1 mL of the dilution was pipetted into a 15 mL centrifuge tube and diluted with 10 mL of 2% aqueous nitric acid solution to detect the iron content by using inductively coupled plasma mass spectrometry (ICP-MS)  . The irop < 0.05.The WHO defines ID as a serum ferritin level less than 15.0 ng/mL and IDA as serum ferritin level less than 15.0 ng/mL and Hb less than 10.5 g/dL. We divided the subjects into three groups according to the WHO definitions: The normal, ID, and IDA groups. All data were confirmed to have a normal distribution by using the Kolmogorov–Smirnov test. Data are presented as means ± standard deviations (SDs), median (interquartile range), or percentage. Intergroup differences were determined using one-way ANOVA, followed by the Scheffé method for post hoc test or nonparametric statistics. Pearson’s chi-squared test was used to assess categorical variables. The correlation between Hb and dietary iron intake was determined using the Pearson correlation test. The association between feeding types and anemia was determined using multivariable logistic regression. All data analyses were performed using SPSS . Differences were considered significant at 3, suspected of infection, were excluded. Data on the iron status analysis of 509 infants are presented in A total of 1368 infants were eligible for this study. However, 779 mothers did not provide consent to extract their infants’ blood, and therefore, the 589 subjects were ultimately enrolled in this study. However, blood draws were unsuccessful in 39 infants. Thus, a total of 550 cases were included for data analysis. Because no introduction of complementary food to infants over six months of age was considered as abnormal feeding, four infants aged eight months and two aged 12 months were excluded accordingly. In addition, 35 infants with a WBC count of >0.000/mmp < 0.001). No significant intergroup differences were observed in terms of the mother’s age, body mass index (BMI), gestational weight gain, and education.Anemia was defined according to the WHO criteria, and 49 (10%) and 21 infants (4%) were diagnosed with ID and IDA, respectively, by the physician . No statThe iron status of the infants and mothers before delivery are presented in The total iron intake was calculated as the sum of the iron contents in milk and complementary food, as presented in The normal group had the highest iron intake from milk (5.04 mg daily), whereas, the ID and IDA groups had a low iron intake from milk. This could be attributed to the continuous breastfeeding of the infants in the ID and IDA groups. No intergroup differences were noted in terms of the iron intake from complementary food. Therefore, milk was the major source of iron intake in infants aged 7–12 months. Furthermore, we determined that the prevalence of ID and IDA in infants aged 7–12 months was 20.4% and 6.6%, respectively, which was significantly higher than 3.7% and 2.7%, respectively, in infants aged 1–6 months. Our results revealed that infants with ID or IDA before six months were all in aged 4–6 months and breastfed. The medium iron intake in infants aged 4–6 months in the normal, ID and IDA groups were 5.13 mg (6.54), 0.13 mg (0.18), and 0.13 mg (0.29), respectively. In aged 7–12 months, infants with IDA had the highest breastfeeding rate, followed by those with ID.r = 0.292, p < 0.001) .Iron is essential for hematopoiesis. We observed that the total dietary iron intake was positively correlated with Hb (< 0.001) A. We fur< 0.001) B and 7–1< 0.001) C. The dap < 0.001). Furthermore, ORs for ID and IDA significantly differed after adjustment for variables in models 2 and 3.Logistic regression models were employed to identify predictors of ID and IDA. Our data revealed that feeding type was a major indicator in predicting anemia . In modeThe period of infancy constitutes a critical window of growth and brain development, and thus, micronutrient deficiencies during this period may have adverse effects on neurocognitive functions. Iron deficiency is the most prevalent nutritional deficiency among infants in developing countries. Women of childbearing age are at risk of iron deficiency because of poor iron content in the diet, increased demand for iron during pregnancy, and iron loss during menstruation and childbirth. In addition, breastfed infants are vulnerable to developing ID because of rapid growth, depletion of their iron endowment, and low iron content in breast milk and in some complementary foods. In our study, among the 509 mother–infant dyads who submitted complete data, 3.7% and 2.7% of the infants aged below six months had ID and IDA, respectively. However, in infants aged above six months, the prevalence of ID and IDA rapidly increased to 20.4% and 6.6%, respectively. Analysis of the maternal hematologic data revealed that anemia was not detected in the mothers during the routine prenatal examination. The basic characteristics of all infants  were similar except for the chronological age and feeding type.The prevalence of ID and IDA varies greatly among countries worldwide. In India, the prevalence of ID in infants aged 3, 4, and 5 months was 5.4%, 21.4%, and 36.4%, respectively, whereas, that of IDA was 4.6%, 16.7%, and 11.4%, respectively . In TurkBreast milk is considered the best food for infants, because it is highly nutritious for infant growth and contains maternal antibodies that provide defense against pathogens. A recent study indicated that human breast milk has very little iron (0.5 mg/L at one month and 0.29 mg/L at 3–5 months) . Our datStudies have indicated that the risk factors for the development of ID included small-for-gestational-age, infants below 10th percentile weight for gestation, infants of diabetic mothers, very-low-birth weight preterm neonates  and infants with lower iron stores ,31. TherAnother possible reason causing iron depletion in infants below six months of age could be maternal iron deficiency during pregnancy, which results in offspring with inadequate iron storage in the body. Some studies have found little or no correlation between maternal and neonatal iron status, whereas, others have suggested that the fetus was vulnerable to maternal ID ,34. MothThe WHO recommends exclusive breastfeeding of infants for the first six months of life. Thereafter, infants should receive complementary foods with continued breastfeeding up to two years of age or above. Taiwan’s health policy also follows this recommendation. In this study, among 328 infants below six months of age, only 17 were introduced to complementary food. Most of these foods were cereals or fruit juices that had low iron content. Thus, in addition to prenatal storage, formula milk contributed more iron to the infants below six months of age. Although human milk has a high bioavailability of iron, the iron content was so low that exclusive breastfeeding increased the risk of ID and IDA. We determined that the total iron intake was positively correlated with Hb level in infants below six months of age, thereby indicating that although prenatal storage was crucial to maintain iron status, additional dietary iron intake can improve the Hb levels and prevent ID and IDA, especially in infants aged 4–6 months. Evidence from randomized control trials suggests that the rate of IDA in breastfed infants could be positively altered by the introduction of solids at four months of age . The AmeAlthough infants aged 7–12 months were provided complementary foods, the iron intake from complementary foods was not different among the normal, ID, and IDA groups. Notably, milk was the major source of iron. Upon further analysis of data, we observed that up to 70% of infants with ID and 90% of infants with IDA were still breastfed. This result indicated that infants who are still breastfed for more than six months do not obtain adequate iron from breast milk. This suggests that mothers who still breastfeed after six months could provide high-quality iron-rich foods to reduce the prevalence of ID and IDA. When we reported the iron status of infants to their mothers, most of the mothers presented their infants had good growth and not aware of the insufficient iron intake and that it increased the risk of ID or IDA. Conversely, infants who used commercial formulas instead of breast milk had better iron status because most commercial formulas are iron-fortified. Future health policies should educate mothers to prepare high-quality iron-rich foods or provide iron supplementation for breastfed infants.The maximum iron requirement for infants aged 4–12 months is approximately 1 mg/kg or 10 mg/day. The average full-term infant requires 8 mg of iron daily from approximately six months of age . Our datAlthough our results determined that breastfeeding was associated with an increased the prevalence of ID and/or IDA, the advantages of breast milk are recognized and unquestionable. Thus, more attention should be paid to the problem of iron deficiency in infants, and strategies should be proposed, including improving maternal iron status, introducing high-quality complementary food early, and even providing iron supplement to breastfed infants. The Pediatrics Association of Taiwan revised the guidelines for breastfed infants in 2016. The content includes the following: (1) Encourage full-term infants to start breastfeeding as soon as possible after birth, (2) Continue to breastfeed until one year of age. After one year of age, the mother may continue to breastfeed the infants, (3) Breastfed infants should be started on complementary foods at 4–6 months of age. If no complementary foods are added after four months, oral iron supplementation should be started at 1 mg/kg/day. Iron deficiency affects the development of cognition, nerves, and behavior, some of which are long-term and irreversible . In thisThe strength of the present work is that it is the first to look at the association between breastfeeding and ID and IDA during infancy in Taiwan. The results can be used as a reference for nutrition policymakers. However, the study had some limitations. First, this was a cross-sectional study that was unable to clarify the effects of longitudinal nutritional status on the development of ID and IDA. Second, the sample size was relatively small, and all participants lived in northern Taiwan, which might limit the generalizability of the results. Therefore, additional large-sample, multicenter studies are required. Third, we only prioritized examining several clinical iron markers that are more likely related to iron status. Fourth, the lack of information regarding the initiation of complementary feeding, that could significantly impact iron status in infants. Nevertheless, further studies are warranted, including those for following up infants with iron deficiency in the first year to analyze whether it affects their cognition function and growth, as well as whether early introduction of high-quality iron-rich complementary foods can improve the iron status of infants.Our data revealed that exclusive breastfeeding increases the prevalence of ID and IDA in infants aged 4–6 months, and prolonged breastfeed above six months can significantly increase the prevalence of ID and IDA. Although iron in breast milk has good bioavailability, its low iron content results in insufficient iron intake in infants and increases the risk of anemia. Hence, health policies should encourage the early introduction of iron-rich complementary foods and educate mothers to prepare high-quality complementary foods to reduce the risk of anemia, especially in infants above six months. In addition, if no complementary foods are added after four months, oral iron supplementation should be started in breastfeeding infants.