Background: Migraineurs are often plagued by sleep disorders. The university student population is high in number and is more vulnerable to migraines. However, no study has analyzed the sleep quality of students with migraine and related contributing factors.Objective: We used the Pittsburgh Sleep Quality Index (PSQI) scale to assess the sleep of migraine patients among college students and to explore the influencing factors of sleep quality.Methods: We performed primary screening for migraine using the ID-migraine screening, and further assessed headache characteristics, sleep, anxiety, depression, and mobile phone addiction in college students with positive primary screening, then diagnosed migraine according to the third edition of the International Classification of Headache Disorders (ICHD-3). Finally, we analyzed the factors influencing sleep quality using Binary Logistic Regression Analysis. Those with scores greater than 5 points on the PSQI scale were believed to have poor sleep quality.Results: The prevalence of migraine was 6.6%. A total of 545 migraineurs were eventually included in the analysis, the incidence of poor sleep quality was 64.04%. The three factors of experiencing aura , anxiety , and high Mobile phone addiction index (MPAI) score  contributed enormously to poor sleep quality. Moreover, the factors of aura symptoms , anxiety , and MPAI score  influenced the sleep quality of female migraineurs rather than male migraineurs.Conclusions: The incidence of poor sleep quality is high among university students with migraine. Aura symptoms, anxiety, and high MPAI score influence the sleep quality of migraineurs, especially females. The proposal of prevention and intervention measures is of great importance to the physical and mental health of students with migraine.Clinical Trial Registration: identifier ChiCTR1800014343. Migraine is one of the most common types of primary headaches with high morbidity and disability, and it is reported that the current prevalence of migraine worldwide is 10% to 18% . Future research applied to the participants who answered “yes,” excluding those who answered “no.”In the second stage, ID-migraine screening was conducted with three simple questions: (1) are there any social, occupational, learning or daily activities affected by headaches in the past 3 months?; (2) do you have a stomach upset, nausea or vomiting during a headache attack?; and (3) are you afraid of light with a headache? ID-migraine is a simple but effective headache screening tool and is often used to diagnose migraine , Headache Impact Test-6 (HIT-6) scale, Hamilton Anxiety and Depression Scales (HAMA and HAMD), Pittsburgh Sleep Quality Index (PSQI) scale, Mobile Phone Addiction Index (MPAI) scale, and others. The research members were made up of professional neurologists in the Affiliated Hospital of North Sichuan Medical College, graduate students and undergraduate students majoring in clinical medicine at North Sichuan Medical College. Before the survey, each member was trained in professional knowledge and questionnaire-related questions and they all passed relevant tests. A unified standard to collect data was also made, insomuch that each member was able to collect data and the collected data were qualified. During data collection, all researchers strictly adhered to the Standard Operation Procedure (SOP) and before the research, they fully understood latent problems and made relevant preparations. the International Classification of Headache Disorders , number of days with headache per month, painkiller use, among others.The degree of headache was assessed using the VAS, with 0 points for no pain, 1–3 points for mild pain, 4–6 points for moderate pain, 7–9 points for severe pain and 10 points for being the most painful  and odds ratio (OR) and all the p-values were two-tailed with a significance level of 0.05.Data analysis was conducted with SPSS26.0 and Forest plotting was conducted with GraphPad Prism8.0.2. Kolmogorov-Smirnov test (K-S test) was used to test the normality of distribution features of measurement data. Measurement data in accordance with normal distribution were expressed as mean ± standard deviation , which had been reported in the previous studies . Sufferers’ headache frequency, HIT-6 score and MPAI score in the poor sleep quality group were higher than those in the good sleep quality group. Likewise, the students in the poor sleep quality group also had higher ratios of anxiety, depression, and aura symptoms. However, in terms of age, sex, Body mass index (BMI), family history, duration of headache, headache intensity, and mobile phone addiction, there are no statistical differences between the two groups (P > 0.05).Of the 545 migraine sufferers included in this study, their average age was 19.59 ± 1.59 years old and 437 of them were schoolgirls (80.18%). The migraine sufferers who obtained more than 5 points from the PSQI scale were classified into the group of “poor sleep quality” and those who obtained 5 or less than 5 points were classified into the group of “good sleep quality.” Analysis of general demographic data, headache features, accompanying psychiatric problems and MPAI situations of migraine sufferers in the two groups were implemented and the detailed results are shown in P > 0.05).MPAI scale survey results: 45 students (17.0%) in the poor sleep quality group spent an average of over 8 h on their mobile phone per day, which is much more than the 17 students (11.5%) in the good sleep quality group. Students with a mobile phone for more than 5 years in the good sleep quality group accounted for 35.8%, higher than those in the poor sleep quality group (32.8%). Students who spent an average of 51–100 yuan on mobile phones per month took up the largest proportion (44.1%) in the poor sleep quality group, whereas their counterparts mostly spent less than 50 yuan (48.7%). As for the main motives for mobile phone use, students in both groups were for interpersonal needs and recreation, with the respective proportions of 23.1% and 49.0% in the poor sleep quality group and those of 26.6% and 48.2% in the other group. However, the aforementioned items made no statistical difference (P = 0.977), sleep latency , sleep duration , sleep disturbance , use of sleeping medication , daytime dysfunction , sleep efficiency , PSQI total score . Migraine sufferers with aura symptoms had a higher average of sleep disturbance than those without aura symptoms who, on the other hand, had a higher score of daytime dysfunctions than migraine sufferers with aura symptoms, both of which were statistically significant (P < 0.05). Though the average sleep duration of migraine sufferers without aura symptoms (6.75 ± 0.937 h) was longer than those with aura symptoms (6.62 ± 1.066 h), the difference was not statistically significant (P > 0.05), see Based on the DC of migraine sufferers with and without aura (MA and MOA) from ICHD-3, students with migraine in this study were classified into two groups, one group being students with aura symptoms and the other group without aura symptoms. The total and sub-item PSQI scores are as follows: sleep quality (P > 0.05) and the sufferers with aura symptoms , anxiety , and depression  were more liable to poor sleep quality. Moreover, migraine sufferers with high HIT-6 scores , headache frequency , and MPAI scores  were more likely to have poor sleep quality.Univariate logistic regression analysis was performed on the items with statistical differences between the two groups in P < 0.05), but sex , HIT-6 score , headache frequency , and depression  had no impact on sleep quality.Such factors as sex, aura symptom, HIT-6 score, headache frequency, depression, anxiety, and MPAI score were included in binary logistic regression analysis as independent variables, in P > 0.05), but they affected the sleep quality of female participants and the effect was of statistical significance (p < 0.05). Factors influencing sleep quality in migraine and female migraine patients are shown in A further binary logistic regression analysis of the factors that influence the sleep quality of male and female migraine sufferers was conducted and the results are displayed in This study reveals that the prevalence of migraine among students is 6.6%, which is lower than that shown by previous studies in China on the general population and students . MA sufferers obtain higher scores in sleep disturbance, whereas MOA sufferers get higher scores in daytime dysfunction. According to Vgontzas et al.  studies of migraine attacks in the aura stage . The results of binary logistic regression analysis showed that aura symptoms, anxiety, and MPAI score were the factors that influenced the sleep quality of migraine sufferers, but we found discrepancies when we further analyzed the factors by sex. For the male migraine sufferers, the risk of poor sleep quality increased by 1.667 times, 1.957 times, and 1.015 times, respectively when they were diagnosed with aura symptoms, anxiety, and mobile phone addiction, but the data were not statistically significant (p > 0.05), which illustrates that for male migraine sufferers, these factors fail to affect their sleep quality. However, for female migraine sufferers, the risk of poor sleep quality increased by 3.796 times, 3.146 times, and 1.028 times, respectively when they were diagnosed with aura symptoms, anxiety, and mobile phone addiction. The risk results were of statistical significance (p > 0.05) and consistent with the overall regression analysis results of students with migraine. What is more, the OR for each factor in the female migraine population was higher than that in the overall migraine population. The discrepancies in contributing factors due to sex suggest that in clinical work, different emphasis needs to be put on the sleep quality of male and female migraine patients, so more targeted treatment plans can be provided.In this study, there was no difference in the sleep quality of male and female migraine sufferers , but we failed to discover the factors that influence the sleep quality of male migraine sufferers, which deserves further exploration. The examination schedules of each semester may affect the quality of students’ sleep, but this factor was not addressed in this study. We did not further describe the type of aura and the proportion of chronic migraine. Moreover, the study failed to further analyze how significant the effect of each factor is, which is also one of the limitations.In conclusion, our study discovers that the prevalence of migraine among students at North Sichuan Medical College is 6.6%, and the incidence of poor sleep quality among students with migraine is up to 64.04%. Such factors as anxiety, aura symptoms, and MPAI score are the risk factors affecting sleep quality, while depression, headache frequency, headache intensity, and duration show no influence on sleep quality. More than 70% of the participants with migraine spend an average of more than 4 h per day on mobile phones, and the avoidance of mobile phone addiction will help improve sleep quality and relieve headaches. Poor sleep quality seriously affects the study and life of students with migraine, so appropriate prevention and intervention measures are of great significance to improve their physical and mental health.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Ethics Committee of Affiliated Hospital of North Sichuan Medical College, and the project number was 2017ER (R) 040. The patients/participants provided their written informed consent to participate in this study.JZ, YC, and JiaL contributed to design the research study, manuscript conceptualization, methodology, writing original draft and translation. JiaL and JW contributed to funding acquisition, project administration and supervision. JY, CL, YL, ZR, and YX contributed to investigation and data collection and curation. JH, JinL, YZ, and WL analyzed the data and provided critical revisions. All authors are accountable for all aspects presented in this study. All authors contributed to the article and approved the submitted version.This work was supported by the Funding Project of the Bureau of Science and Technology and Intellectual Property of Nanchong City (No. NSMC20170420 and No. 19SXHZ0023).


Skeletonema grevillei cultivated in a bioreactor and an incubation-shaking cabinet at different growth phases (after 192 and 312 h). Growth was monitored by evaluating cell density with the Sedgewick Rafter chamber, and the collected biomass was extracted with 70% ethanol assisted by ultrasound. Extracts were evaporated to dryness and compounds were identified in derivatized form by gas chromatography and mass spectrometry (GC-MS) analysis, while antioxidant capacity was evaluated by DPPH and ORAC. Significantly faster growth was observed in the bioreactor than in the incubation-shaking cabinet. Oleamide, palmitelaidic acid, glycerol monostearate, myristic acid, cholesterol, eicosapentaenoic acid, 1-monopalmitin, and 24-methylene cholesterol were identified as the major compounds in both systems. Among them, oleamide was the dominant compound in both systems. It is also shown that prolonging the cultivation period had a direct effect on increasing the extract yield. The highest DPPH inhibition (11.4 ± 1%) and ORAC values (93.3 ± 8.4 mM TE) were obtained for the S. grevillei extract recovered from the bioreactor after 312 h. The obtained results contribute to the possibility of using S. grevillei for various biotechnological applications in the future.Marine microalgae, diatoms, are considered a source of a wide range of high-value compounds, and numerous studies indicate their biotechnological potential in the food and feed industry, cosmetic industry, nanotechnology, pharmaceutical industry, biodiesel production, fertilizers, and wastewater treatment. The aim of this study was to compare the growth, chemical profiles, and antioxidant activity of the diatom  Market demand is changing rapidly, directing research toward finding new ways to use marine resources as a renewable and sustainable source of compounds that can be used in a variety of industries. Among the great biodiversity found in the oceans and seas, marine microalgae are recognized as microorganisms with exceptional biotechnological potential, especially in industries directed to production of pharmaceuticals, functional food ingredients, and food products . AlthougDunaliella salina. As the biotechnological potential of diatoms is constantly being discovered, the number of studies on different species is increasing. Diatoms have shown promising features that make them ideal candidates for this form of production, as their intracellular content can be used for the production of biodiesel and valuable food components, and the silicate shell can be used as a material for nanotechnology [Skeletonema is an unconventional group of microalgae, but since it is non-toxic, it is suitable for use in the food industry [Skeletonema sp. as generally recognized as safe (GRAS) [S. marinoii have been described in the literature [S. grevillei. Nevertheless, the cultivation of microalgae for wider use slowed down due to technological obstacles, mainly related to the optimization of growth, harvesting, and extraction efficiency of targeted compounds [Diatoms are photosynthesizing microalgae with cell walls of transparent opaline silica containing compounds such as pigments, sterols, and fatty acids . Currentchnology . In termindustry . In facte (GRAS) . Some spterature ,8; howevompounds . In addiNitzschia sp. In general, cultivation in bioreactors achieves greater homogeneity so that all cells have the same availability of key factors for successful growth, such as nutrients and light. While the production of higher amounts of primary metabolites is associated with optimal growth conditions, the accumulation of secondary metabolites is usually associated with stressful conditions [Skeletonema. Some species such as S. marinoii, S. dohrnii and S. costatum have been previously studied [S. gravileii. In a nutrient-deprived environment, Skeletonema species are able to increase lipid production [Diatoms are among the most flexible and productive eukaryotic microalgae characterized by exceptional adaptability to changing environmental conditions . Mechanisms such as extracellular polysaccharide layers, cell wall thickness, and morphologically distinct resting stages minimize the potential damage caused by stress conditions ,11,12. Onditions . Microalnditions ,15. Neve studied ,16,17,18oduction , and eveoduction . Under loduction ,21. In aS. grevillei in a standard F/2 medium using two cultivation systems (bioreactor and incubation-shaking cabinet). In addition, ethanolic extracts were prepared for each system at two growth stages and their chemical profiles (dominant compounds) and antioxidant potential were determined.Biotechnological improvements are possible by selecting suitable microalgal strains capable of producing large amounts of various bioactive compounds under optimal conditions. Such a form of microalgae cultivation would make commercialization faster and more profitable. Therefore, the aim of this study was to determine and compare the growth of Considering that extending the duration of the cultivation period beyond the stationary phase would result in the decay of diatom cells, the end of cultivation in this study was defined as the time at which one of the systems reached maximum growth. This was considered the beginning of the stationary phase and in the bioreactor system (BRC) it was reached after 312 h.p < 0.05) change  was observed in the EIS system, while in the BRC system this was observed one day earlier. During the first 144 h, there was no statistically significant difference (p > 0.05) in cell number between the EIS and BRC systems. A statistically significant (p < 0.05) difference between the systems was first observed after 168 h and persisted until the end of cultivation. The total mass of dry extracts collected in the exponential and at the beginning of the stationary growth phase is shown in The results of growth in a BRC and the incubation-shaking cabinet (EIS) system during the cultivation period are shown in N. epithemioides and Nitzschia sp. at different temperatures, light intensities and silicate levels. The addition of silicate was found to be more suitable to stimulate growth than modification of light and temperature. Raniello et al. [Cocconeis neothumensis under different light and nutrient conditions in a bioreactor than in a batch culture in Petri dishes. Significantly higher extract yield (24 ± 5 mg of dry extract) was obtained in the bioreactor compared to batch cultures (17 ± 3 mg of dry extract), which is comparable to the results of extract yield between BRC and EIS systems in this study [Since the early days of diatom cultivation in bioreactors, it has been known that cell homogeneity allows maximum utilization of limiting nutrients . Althougo et al.  recordedis study .m/z of the compounds is given in the The compounds identified in a BRC, and EIS are listed in Z)-9-octadecenamide) was dominant in both cultivation systems and at both phases. After 192 h, the percentage of oleamide in BRC and EIS was 49.84% and 73.31%, respectively. In both systems, there was a significant decrease with the extension of the cultivation period, by 16.19% in BRC and 37.78% in EIS.The difference in the number of compounds identified in the BRC system after 192 and 312 h of cultivation was 44 and 42 compounds, respectively. Compared to BRC, a lower number of compounds were identified in both phases of the EIS system, 33 after 192 h and 34 after 312 h of cultivation. Oleamide  while the increase was lower in BRC at the end of the cultivation period (from 5.92% to 9.30%). To our knowledge, this is the first time it has been identified in marine diatoms. In both systems, myristic acid increased by 3.95% in EIS and 3.53% in BRC, with an additional culture period of 120 h. After 312 h of cultivation, EPA was identified in both systems but the proportion in BRC was higher than in EIS, 6.40% and 3.78%, respectively. In addition, after 192 h, EPA was not detected in the EIS, while in BRC, it was recorded in proportion of 3.29%. The proportion of 1-monopalmitin in BRC remained almost unchanged (from 2.91% to 2.89%), and the same compound showed a sharp increase in EIS (from 0.47% to 6.92%) with the extension of cultivation.In addition, an increase in tridecanoic acid, loliolide, pentadecanoic acid, palmitic acid, phytol, 2-monostearin, and loliolide was observed in both systems over cultivation time, while the proportion of decanoic acid remained unchanged. At the end of the cultivation period, some newly identified compounds in a proportion of less than 1%, were malic acid, (Z)-3-hexenyl-β-glucopyranoside, palmitoleamide, 1-octadecanol, desmosterol in BRC and butanedioic acid, hexadecanenitrile, 1-hexadecanol in EIS.There were eight compounds that were predominant in both systems. These are listed in Chromochloris zofingiensis and in few terrestrial plants. There are numerous biological activities related to this compound [Ipomoea and Dillenia species) excretes anti-inflammatory properties [Colletotrichum gloeosporioides has confirmed antimicrobial activity against Staphylococcus aureus (zone of inhibition = 25 mm) [Diaporthe schini, in which oleamide was a dominant metabolite, also showed antimicrobial activity against S. aureus (MIC = 125 uL/mL) [Aspergillus flavus and Klebsiella pneumonia [Microcystis aeruginosa NIES -843, a toxin-producing cyanobacterium [Tetraselmis tetrathela when cultivated at a salinity of 30 ppm [Oleamide is an amide derived from oleic acid. It is found in green algae such as compound ,30,31. Icompound . It was operties . Further= 25 mm) . The ext5 uL/mL) . Furtherneumonia . It is af 30 ppm .S. grevillei for both cultivation systems. Marzec et al. [Halamphora cf. salinicola (strain SZCZM1454A). Myristic acid has been described for a variety of biological activities, including antifungal, antiviral, anticancer, and antiparasitic activities [Fatty acids are known for their antimicrobial and antibiofilm activity. Monounsaturated palmitelaidic acid showed antibiofilm activity against Escherichia coli, inhibiting 25% of biofilm formation at 256 µg/mL, while inhibition of S. aureus was 21% at a concentration of 16 µg/mL . In genec et al.  also foutivities .S. grevillei we observed the decrease of 1-monopalmitin, probably due to the synthesis of fatty acids such as eicosaonic, EPA, and 5,8,11-eicosatrienoic acids.An important cellular metabolite in triacylglycerol (TAG) biosynthesis is 1-monopalmitin. When TAGs are synthesized more intensively, a decrease in the content of 1-monopalmitin can be observed . In S. gDichapetalum filicaule and reported for antihelminthic activity against hookworms [Glycerol monostearate is known as the glycerol ester of stearic acid. There is no available literature on this compound from diatoms. So far it has been extracted from plant ookworms . In geneS. grevillea, cholesterol, desmosterol, and isofucosterol. In addition to cholesterol, 24-methylene cholesterol was found among the dominant compounds. The proportion of 24-methylene cholesterol in BRC was 3.44% higher than in EIS at the end of the cultivation period. The mass spectra of the fourth sterol compound, 24-methylenecholesterol in derivatized form , plants (24-methylene cholesterol) and alga (fucosterol) were previously identified in the diatom species S. marinoi [S. marinoi strain does not possess genes for lanosterol synthetase, but that desmosterol and cholesterol are produced via cycloartenoid pathways [S. marinoi belongs to the same order as S. grevillei, it can be hypothesized that the biosynthesis of sterols occurs in a similar manner.These results are consistent with previous studies that characterized the genus  marinoi . Cultiva marinoi . The mod marinoi . A simil marinoi ,50,53. H marinoi  confirmepathways . Since SThe results of antioxidant activity (DPPH and ORAC) of diatom extracts are shown in In addition to the DPPH method, the ORAC method was chosen because it best simulates in vivo conditions and provides results for antioxidant levels equivalent to those in biological systems . For theS. grevillei extracts also increased. A statistically significant difference (p < 0.05) in antioxidant activity was detected between cultivation systems at the same growth stage for ORAC, but not for DPPH. Fatty acids and sterols are recognized as compounds with strong antioxidant activity ,55. The Tetraselmis sp. IMP3, Tetraselmis sp. CTP4, and Skeletonema sp. was evaluated using the methods ferric ion reducing antioxidant power (FRAP), DPPH, and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) [Skeletonema sp. while the results of ethanolic extracts obtained with the DPPH method were comparable to those obtained in the present study.Antioxidant activity of aqueous and ethanolic extracts of marine microalgae ) (ABTS) . The higLiCrosolv Ethanol (gradient grade), Sigma Aldrich  was used for extraction. In GC/MS analysis NO-Bis(trimethylsilyl)trifluoroacetamide (BSTFA) from Sigma-Aldrich  was used as a derivatization reagent while dichlormethane  was used as a solvent for the non-derivatized sample.S. grevillei (CIM876), isolated from the northern Adriatic Sea, was donated from the culture collection of the Center for Marine Research of the Ruđer Bošković Institute . The diatom was cultured in F/2 medium in two parallel systems: (i) in a bioreactor (BRC) , (ii) in a 5-L Erlenmeyer flask set in an incubation-shaking cabinet (EIS) . Two replicas were used for each system, each containing 2 L of F/2 medium inoculated with 50 mL of S. gravillei (9 × 104 cells/mL). The medium was prepared according to the previously described recipe [The strain of d recipe . In both systems, diatoms were cultured at 18 °C with a light–dark photoperiod set to 12 h. The light intensity was 7500 lux in both cultivation systems. In BRC, a LED light  was used (a combination of cool white and blue), and fluorescent lamps  were used in EIS. In BRC, the diatoms were stirred with an impeller at 70 rpm and the air flow rate set at 1 L/min, while in EIS they were shaken at a low speed (50 rpm) to avoid spillage.To compare the growth rate in BRC and EIS systems, 5 mL of the cultures were collected every 24 h of the cultivation period under sterile conditions. The growth curve was determined by counting the cells in a Sedgewick Rafter chamber using a light microscope  at 200xX magnification . g, at 4 °C for 30 min to separate the silicate frustule. The collected supernatant was filtered through a sterile CA filter with a pore diameter of 0.2 µm  and evaporated in a centrifugal evaporator  until completely dry. The mass of the dry extract was recorded.The biomass was collected at the exponential and at the beginning of stationary growth phases. These were determined at 192 and 312 h of culturing. Using the filtration glass microfiber filters (Grade GF/F Whatman) at 3.12 psi pressure the biomass was collected and transferred to falcon tubes and re-suspended in 10 mL of 70% ethanol. Extraction was performed in duplicate by ultrasound-assisted extraction (UAE) using an ultrasound probe  set at 20 kHz ± 500 Hz. The extraction was performed for 5 min, at an amplitude of 30% and treatments of 20 s. The break between treatments was 40 s. The samples were centrifuged  at 29,097× The identification of compounds was done according to Torras-Claveria et al. with minor modifications . BrieflySince commercial libraries did not offer adequate mass spectra for the identification of compounds in derivatized forms, the same samples were analyzed in non-derivatized form as well. The analyses were performed according to the same protocol, but 50 μL dichloromethane was used instead of the derivatizing agent. The derivatized and non-derivatized compounds were identified by their retention index and mass spectra using the Wiley 12 & NIST 2020 database.Prior to analyses, dried extracts were dissolved in 70% ethanol at a concentration of 10 mg/mL. Two assays, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and oxygen radical absorbance capacity (ORAC) were used to determine the antioxidant potential of the extracts.DPPH radical scavenging ability of diatom extracts was measured in 96-well microplates, following the method previously reported by Čagalj et al. . Brieflyp < 0.05) in the growth rate in each system over time and the difference between the EIS and BRC systems, as well as antioxidant activities [Cell density data, collected and recorded using the Sedgewick Rafter method, were used for the analysis. The analyses of variance (one-way ANOVA followed by Fisher’s least significant difference test) was used to determine the significant changes (tivities . AnalyseS. grevillei showed low antioxidant activity at the tested concentration, so further studies should focus on the biological activity at higher concentrations and quantitative determination of the biologically active compounds. It should also be emphasized that, in addition to the extracts, silicate diatom frustules are also used in nanotechnology, which will certainly affect the profitability of the production of these microalgae in the future. Future research should focus on finding conditions under which it is possible to maximize the production of metabolites that have the potential for biotechnological applications.This study confirms that bioreactor cultivation is a successful system for the cultivation of diatoms. Optimal growth conditions in bioreactors allowed faster entry into the exponential and stationary growth phases. It could be hypothesized that prolongation of cultivation would result in slightly higher biomass in the bioreactor; however, the prolongation would also result in the death of diatom cells. The results of the chemical profiles revealed that oleamide was most abundant compound in the BRC and EIS systems, and that palmitelaidic acid, glycerol monostearate, myristic acid, cholesterol, eicosapentaenoic acid, 1-monopalmitin and 24-methylene cholesterol were also abundant. All of the mentioned compounds have been previously characterized for their biological activities, which is why some of them are already used in industry, while the other compounds have the potential for biotechnological applications. Moreover, in this study, tentative identification of derivatized 24-methylene cholesterol was obtained, which was not previously found in its derivatized form in the commercial libraries but is known from the literature. For precise identification techniques such HR-MS or NMR should be used. In addition, ethanolic extracts of


Aedes aegypti is the main vector that transmits viral diseases such as dengue, hemorrhagic dengue, urban yellow fever, zika, and chikungunya. Worldwide, many cases of dengue have been reported in recent years, showing significant growth. The best way to manage diseases transmitted by Aedes aegypti is to control the vector with insecticides, which have already been shown to be toxic to humans; moreover, insects have developed resistance. Thus, the development of new insecticides is considered an emergency. One way to achieve this goal is to apply computational methods based on ligands and target information. In this study, sixteen compounds with acceptable insecticidal activities, with 100% larvicidal activity at low concentrations (2.0 to 0.001 mg·L−1), were selected from the literature. These compounds were used to build up and validate pharmacophore models. Pharmacophore model 6  was used to filter 4793 compounds from the subset of lead-like compounds from the ZINC database; 4142 compounds  were then aligned to the active site of the juvenile hormone receptor Aedes aegypti (PDB: 5V13), 2240 compounds  were prioritized for molecular docking from the construction of a chitin deacetylase model of Aedes aegypti by the homology modeling of the Bombyx mori species (PDB: 5ZNT), which aligned 1959 compounds , and 20 compounds  were predicted for pharmacokinetic and toxicological prediction in silico . Finally, the theoretical routes of compounds M01, M02, M03, M04, and M05 were proposed. Compounds M01–M05 were selected, showing significant differences in pharmacokinetic and toxicological parameters in relation to positive controls and interaction with catalytic residues among key protein sites reported in the literature. For this reason, the molecules investigated here are dual inhibitors of the enzymes chitin synthase and juvenile hormonal protein from insects and humans, characterizing them as potential insecticides against the Aedes aegypti mosquito. Aedes aegypti mosquito, the main transmitter of arboviruses such as dengue, dengue hemorrhagic fever, urban yellow fever, and chikungunya  (n) + n H2O = n acetate + chitosan -3,7-dimethylnona-2,6-dienoate). Therefore, 2240  compounds were chosen since the efficiency of the ligand selects a more concordant compound for the choice of a principal compound [The molecular adjustment was performed using the AutoDock Vina program, and all parameter values were adjusted throughout the process to define receptors, exhaustiveness, center_xyz, and size_xyz. To generate the result in binding affinity values, the AutoDock Vina program foresees the connection of the lowest energy state at the time of the formation of the protein complex (receptor) with the ligand, with the objective of a correlation between the values of the functions and the affinity of the ligand ,53,54,55compound ,57,58.JH3) within the enzyme active sites (PDB ID 5V13). The root mean square deviation (RMSD) between the reference ligand and the experimental JH3 ligand (1.19 Å) was calculated. The RMSD analysis was determined by considering the most appropriate position of the initial structure around the X-ray crystallographic complex through an experiment of the most stable position so that there is no structural change between the protein and the reference ligand when complexed with the macromolecule under study. The best molecular fit is determined by an RMSD less than or equal to 1.5 [Molecular docking was performed using a validation protocol to determine the pose (conformation + orientation) of the ligands  was carried out on all compounds, and, after filtering the ligand efficiently, 2240 compounds went on to the next stage of molecular docking with the model Bombyx-Aedes enzyme insect chitin deacetylase built from the The juvenile hormone protein presents in its binding site the main interaction with the epoxy group and forms a hydrogen bond with the phenolic hydroxy group of Tyr129; the rest of the isoprenoid chain is surrounded by hydrophobic side chains, including those of Phe144, Tyr64, Trp53, Val65, Val68, Leu72, Leu74, Val51, and Tyr33 ,65.Aedes aegypti juvenile hormone macromolecule (COD: 5V13). All selected ligands (M01–M05) and the reference ligand complexed with the macromolecule (JH3) interacted with residues Tyr64 (except M03), Trp53, Trp50 (except M04), Try33 (except M03 and M05), and Val51 (except M03 and M04). M03 was the only one that interacted with the amino acid residues Val65 and Val68, as shown in When we analyzed our ligands, important interactions were carried out with the main amino acid residues of the binding site of the Aedes aegypti mosquito. In view of this approach, 2240  compounds were selected in the first stage with the juvenile hormone macromolecule. For the next stage, there was, again, a selection of the molecular docking based on the insect model Aedes aegypti chitin deacetylase, constructed from the enzyme insect chitin deacetylase deposited in the PDB (PDB code: 5ZNT); 1959 compounds  were aligned and 20 compounds  were prioritized for pharmacokinetic and toxicological prediction.The selection of compounds took place in a double inhibition of macromolecules, where the parameter to which they were associated performed more satisfactorily in filtering the choice of the most appropriate ligand that would, in particular, function as both an inhibitor of the juvenile hormone and a growth regulator of the When we analyzed the interactions described by Liu , the 5ZN7 (residues 162–471) composed of seven parallel β strips arranged in a barrel surrounded by six α helices. The active site and the substrate connection gap are located through the upper center of the barrel (β/α)7, which contains a metal bond triad conserved throughout the CE4 family, namely, a zinc ion coordinated by Asp206 and Asp205, His261 and His265, in addition to Arg306 and canonical Arg306, responsible for stabilizing the catalytic base Asp205 [In addition, the chitin deacetylase enzyme catalyzes the removal of acetyl groups from chitin and modifies this polymer during its synthesis and reorganization. The enzyme has a structure of two regions: a subdomain of NodB homology conserved in CE4 and C-terminal loops. The homology domain of NodB is a barrel (β/α)e Asp205 ,42.silkmoth Bombyx mori (BmCDA1-CAD), with only one hydrophobic residue exposed to the solvent (Tyr242) [A long crack in the substrate was also observed on the surface of the insect chitin deacetylases (CDAs) from the (Tyr242)  that canM01–M05) and zinc with the amino acid residues His256, His261 (except M01), and Tyr305 (except zinc) were observed. Only compounds M01 and M05 interacted with the amino acid residue Asp302, and compounds M01 and M03 were the only ones that interacted with His430 and Leu306. Compounds M01, M04, and M05 interacted with Ala303, as shown in When analyzing the ligands with the best scores, interactions of all ligands  [g links) ,79.M01–M05, as seen in M03 were the only ones that did not violate any of the parameters, the pharmacokinetic properties of which remained within the range of the graph. The positive control FCX, on the other hand, violated the polarity parameter, making it a very bad insecticide to achieve high concentrations in the target receptors.We checked the radar bioavailability graph, which has six axes of important bioavailability properties for the absorption of the insect’s body wall and CNS. Of the positive controls DFB, TEF, FCX, and BUP and compounds M01, M02, M04, and M05, as per 3 hybridization fraction greater than 0.25, demonstrating that the design of the compound substituents could increase the complementarity of receptors/ligands, making it possible that additional protein–ligand interactions would not be accessible to a flat aromatic ring and, therefore, would improve potency and selectivity for a given target, which should mitigate the off-target effects [However, DFB, TEF (positive controls),  effects .M01, M02, M04, and M05  [Therefore, according to the literature, the greater the hydrophobic property, the greater the logs (MDCK) ,85,86,87The pharmacokinetic properties are directly related to the proper planning of the insecticidal potential since different substituents on a given chemical entity are used as references to potentially create more efficient compounds according to their hydrophobic, steric, and electronic characteristics. Therefore, when analyzing our compounds, we take into account the octanol–water partition coefficient to define the hydrophobic character of a substituent  on diffeThe SwissADME program for better visualization allows the BOILED-Egg method of estimated brain or instinct permeation, an intuitive method to simultaneously predict two main parameters of the ADME, that is, passages across membranes and brain access (BBB). Although conceptually very simple, as it has only two physical–chemical descriptors , this classification model was constructed with extreme care for statistical significance and robustness ,79,89.M01–M05 was based on the structural simplicity of the derivatives and the fact that it allows the study of their effects on the cuticle and CNS. Since solubility is one of the main physicochemical properties for the development of a drug, the partition coefficient between n-octanol and water in its logarithmic form (log Po/w) could determine the assessment of the pharmacological properties of absorption, distribution, metabolism, excretion, and toxicity (ADMET) of a candidate chemical lead for its initial selection. It establishes benchmarks against which compounds synthesized during lead optimization can be evaluated [The selection of compounds valuated ,91.o/w, processed by the SwissADME program, as can be seen in o/w. This parameter is the arithmetic means of the values predicted by the five proposed methods based on free solvation energies in n-octanol and water, calculated by the generalized-born model, and can give an estimation of the solvent-accessible surface (GB/SA) [o/w of 5.37); however, this compound can overcome the gastrointestinal barrier, which in the insect’s structure would be the passage through the cuticle. However, it cannot overcome the blood–brain barrier in sufficient concentrations due to its atomic nature, with three aromatic rings, together with a cyclopropane, making it possible to increase the log Po/w [Log P (GB/SA) ,94,95,96log Po/w ,98,99.M01, M02, M03, and M04, are found in the yellow region (yolk) [M05 are in the white region of HIA , corroborating previous data. When performing the in silico experiment in the SwissADMET program for the 20 compounds, the program did not generate results for compounds ZINC95851157 and ZINC12447446. They cannot be seen in the graph because their results are out of reach, which can be explained by the fact that they have deficient pharmacokinetic properties, similar to the BCR-ABL-1 inhibitor. They are predicted with an accuracy of 83% (true negatives are out of range) [From the result of n (yolk) . On the f range) .M05 did not cross the blood–brain barrier in a high concentration, in spite of our plan for the growth inhibitors’ possible mechanism by inhibiting the enzyme chitin synthase present in the cuticle. Another analysis parameter in the graph is the possibility of active efflux from the cell to expel the compound that passes through the barriers that are substrates or non-substrates of the permeability glycoproteins PGP+ (substrates) and PGP− (not substrates). Blue and red dots symbolize compounds; none of the compounds were PGP+, shown as blue and red dots on the graph, and none of the compounds were PGP+. Probably they prevent the intracellular accumulation of toxic compounds, as they are eliminated outside the cell through active efflux through biological membranes [This is a clear indication that these compounds are highly likely to be absorbed by the gastrointestinal tract and permeate the insect’s brain; the compounds TEF, FCX, and embranes ,92,93.Aedes aegypti mosquito shows similarity in its identity to other species of insects and to humans [− since this depends on the species of insects, the structural components of the compounds, and how they are analyzed [The glycoprotein-P of the o humans . It was o humans . Howeveranalyzed ,96,97,98Aedes aegytpi juvenile hormone protein, it is expected that the insecticide requires a minimum concentration for the desired effect. A toxicity assessment in humans is essential for any insecticide [http://xundrug.cn/moltox, accessed on 30 June 2021).To design an insecticide that is developed based on certain mechanisms of action, be it the chitin synthase enzyme or the ecticide . Shown iEMolTox is a web server for predicting potential toxicity associated with the structure of a compound; it predicts the compound with high confidence using the aggregate predictor method (ACP). It was elaborated based on literature data from a total of 174 in vitro/in vivo experimental data sets related to toxicology. The data were used to build the model through physical–chemical descriptors and Mondrian’s shaped prediction, which offers similar prediction accuracy for both active and inactive sets, where the validity values are around 0.90, at a significance level of 0.1. This toxicological web server predicts the molecular toxicity of the target molecule based on the mechanism of action in different animal models and their organs ,109,110.M04 compound was on alert for skin injury, together with alerts in the liver for the action of blocking the (OATP1) B1 transporter, which is the same for the M02 and M03 compounds. The organic anion transporter polypeptide (OATP) 1B is a detoxifying limiting protein that mediates the uptake and elimination of many endogenous and xenobiotic compounds (drugs or insecticides) through the sinusoidal membrane of liver cells [M02, M03, and M04 are possibly blocking this transporter, making them molecular structures to be eliminated by living beings. M01 and M05 did not show any actions or injuries , at the request of the PRASA Unit of the European Food Safety Authority (EFSA), on the genotoxic and carcinogenic potential of buprofezin in the context of human risk assessment. It was evaluated in a two-year study that, in mice and rats, neither the nature nor the incidence of tumors was affected by its administration . HoweverN-methyl-4-chloroaniline, of which the last three are classified as mutagenic [Plectonema boryanum) that when the phenylurea derivatives were exposed and became highly resistant, the main metabolite was p-chlorophenylurea [M01–M05 compounds , these compounds could give rise to a few adverse reactions. Teflubenzuron (TEF) is used in the delousing of salmon farms; analyses in non-target species such as lobster (Homarus gammarus) and shrimp  have shown exoskeletal abnormalities (deformities of the claws or rigid legs); deformities of juvenile lobsters have been observed in several organs [The compound that showed excellent results for the standards established by the program when analyzing toxicity ,123 and l organs ,125,126.Insect growth inhibitors possess the basic chemical structure of benzoylphenylurean compounds, which consist of three regions for SAR analysis: the benzoyl ring (A), the urea bridge (B), and the aniline (C), as shown in M01 presented urea in its molecular structure. M01 and its derivatives have 19 studies and patents that are mainly related to antibacterial tests, nitric oxide, DNA inhibition, neuroinflammatory treatment and autoimmune diseases, ribonucleotide reductase, tyrosinase inhibitors, and sickle cell anemia treatments [After the high-performance pharmacophoric screening of the studied compounds, only eatments ,133,134.M05, has 26 studies related mainly to histone deacetylase, urease, metallo-β-lactamase inhibitor FEZ-1, antitumor activity, DNA synthesis, hydroxamic β-glucosyl-phenolic acid inhibitor, ribonucleotide reductase, 5-lipoxygenase inhibitors, and activity against trypanosome [M02 compound has only one study analyzing antibacterial activity [M03 and M04 compounds, there are no biological studies; none of our compound cells were tested against insect-growth-inhibiting activity to transform them into a new insecticide after additional tests in vitro and in vivo.Another widely studied compound, panosome ,142,143.activity ; in relaM01–M05, after hierarchical virtual screening, were submitted for evaluation of synthetic accessibility  in EtOH and H2O as solvents and using NaOH as a base will allow us to obtain the final butyraldehyde oxime—M01  of intermediate V, final hydroxylamine M02 will be obtained [We propose a theoretical synthetic route for nditions . By a reobtained  oxyl (TEMPO), InCl3, and acetaldoxime as catalysts, using toluene as a solvent  hydroxylamine M04 [We present a synthetic route to synthesize compound M04  based onmine M04 .M05 using intermediate XIV as the key substrate. The initial strategy to obtain XIV consists of two steps [XII with o-(tetrahydro-2H-pyran-2-yl) hydroxylamine (NH2OTHP) in DMF with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N,N-diisopropylethylamine (DIPEA) will provide derivative XIII. Then, the cleavage of the THP-protecting group in acidic conditions will produce benzamide XIV. The second possibility of synthesizing the key intermediate can be through palladium-catalyzed hydrogenation [XV. Finally, the target compound M05 can be furnished from intermediate XIV by a hydrogenation reaction using platinum/ceria-zirconia mixed oxide (Pt/CeO2-ZrO2) as a catalytic system [We proffer a theoretical synthetic route for compound wo steps . Firstlygenation , removinc system  , was employed to build ten pharmacophore models from a training set , from which we selected the most reliable tautomers (pH = 6.5) for all compounds [TM 2.0 package (Tripos Inc.). All structures were energy-minimized using a conjugate gradient (CG) with a convergence criterion of 0.001 kcal/mol and a Tripos force field  [TM (Tripos Inc.).First, they were drawn using Marvinompounds . Next, 2 50,000) . Partial 50,000) , availab−1) were selected, seven of which were used to determine the training set and seven for the test set. They were used randomly in the construction of the pharmacophore models, while the other compounds with the same criteria were used in the validation of the pharmacophore model.The study of larvicidal activity was carried out by Sun  et al., TM module in SYBYL-XTM 2.0 [The training datasets were flexibly aligned to each other in order to build hypermolecular alignments that map common pharmacophore features. The genetic algorithm employed in this step starts with 45 conformations (population size) of each benzoylphenylurean derivative and evolves through a maximum of 90 generations by means of default genetic operators (mutation rate = 0.2 and crossover rate = 0.2) available in the GALAHAD-XTM 2.0  , [TM program v. 12.0  by calculating the area under the receiver operating characteristics curve (AUC-ROC curve); it was possible to visualize the recognition of assets and decoys for better validation of the models through the true-positive rate versus the false-positive rate.Pharmacophore models with the lowest energy  were selected and analyzed for the results of the Pareto score. To determine the best pharmacophore model and its discriminatory power, the models were used to build decoys that were removed from the DUD-E server, March (h 2021),  and execwww.jyu.fi/rocker, accessed on 15 March 2021). Additionally, BEDROC is oriented by a search algorithm (α = 16.10) that corresponds to the initial 10% of the database. The pharmacophore model that demonstrated early recognition (BEDROC > 0.5) was employed in the virtual screening step.The Boltzmann-enhanced discrimination of the ROC curve (BEDROC) was employed to discriminate models with AUC > 0.7 according to early recognition of actives using the ROCKER server  . Compounds were ranked according to QFIT values, and those with values higher than mean plus two times standard deviation (Equation (1)) were selected for molecular docking.The best pharmacophore model was automatically converted to the UNITYdatabase  : Mathematical equation of the mean plus two times the standard deviation.Aedes aegypti species was collected in the National Center for Biotechnology Information (NCBI), with the descriptives (Aedes aegypti + chitin deacetylase 1 protein) at https://www.ncbi.nlm.nih.gov/protein, accessed on 27 June 2021. We found that the protein AAEL003419-PA was used as a primary protein; the BLASTTM, https://blast.ncbi.nlm.nih.gov/Blast.cgi, accessed on 27 June 2021 [The primary sequence of une 2021  performeAedes aegypti, performed the comparison, alignment, construction, and validation of the model, called Bombyx-Aedes.The most similar macromolecules were introduced and analyzed in the MODELLER 10.1 program, which, with the primary structure of https://swissmodel.expasy.org/assess, accessed on 27 June 2021) was employed to analyze the model created for A. aegypti chitin deacetylase using the global model quality estimation (GMQE) score on the accuracy of the constructed model and the model used [Using the SWISS-MODEL server ,166,167,del used ; the pridel used . Furtherdel used . In the The conformational search and scoring evaluation were performed using AutoDock Vina 1.1.2 . The ability to generate satisfactory solutions to scoring functions was probed according to the root mean square deviation value (RMSD < 2Å) ,172.Aedes aegypti juvenile hormone receptor (PDB: 5V13) were performed according to previous studies [A. aegypti chitin deacetylase receptor model.The compounds prioritized in pharmacophore-based virtual screening were employed in molecular docking, and in silico approaches of the  studies ,174. AftLE = Ligand efficiencyΔG = Binding affinityN = Number of heavy atomsEquation (2). Mathematical equation of ligand efficiency.A. aegypti chitin deacetylase receptor, and compounds with LE < −0.4 kcal/mol were prioritized for pharmacokinetic and toxicological analysis.The ligand efficiency (LE) was calculated for compounds employed in docking with the https://preadmet.bmdrc.kr/, accessed on 27 June 2021) and SwissADME tools [http://www.swissadme.ch/index.php, accessed on 27 June 2021). The analysis of the toxicity profile of the compounds was evaluated using the online platform eMolTox .For the analysis of pharmacokinetic properties, the best compounds from the previous step were selected, and we used the Preadmet , which implements the tautomer generations by the algorithm method of combinatorics, exhaustively calculating all tautomeric forms of a given organic compound by molecular and stereochemical complexity and considering the presence of fused and bridged systems to suggest the canonical tautomer by an energy base of classification by different structural and topological characteristics combined with an additive scheme using weighted molecular descriptors. By generating a score ranging from 0 to 100 (maximum synthetic accessibility), the molecule is more easily synthesized when the score is closer to 100 [The prediction of the synthetic accessibility (SA) of the compounds selected by the hierarchical virtual screening was performed using the AMBIT program  of the physiology of the Aedes aegypti mosquito, and toxicological properties directed at other organisms, mainly humans, as well as the synthetic theoretical routes of compounds M01, M02, M03, M04, and M05 have been proposed.This study has established the planning of insect-growth-inhibiting compounds with potential insecticidal activity that have the possible mechanism of action of dual inhibition of the juvenile hormone protein; the study has also established a chitin deacetylase model of M01–M05 were selected, showing parameters similar and superior to those observed for positive controls and also significant differences in the interaction with key residues in each catalytic protein site reported in the literature. Accordingly, the molecules investigated here are dual inhibitors of the enzymes chitin synthase and juvenile hormonal protein (from insects), characterizing them as potential insecticides against the mosquito Aedes aegypti. With these promising results, our research group intends to carry out biological tests in vivo with the compounds obtained in this work to confirm the in silico predictions.Finally, compounds


Lately, concern was expressed about the possibility that gases from respiration could build up in the mask over time, causing medical issues related to the respiratory system. In this research study, the carbon dioxide concentration and ethylene in the breathing zone were measured before and immediately after wearing surgical face masks using the photoacoustic spectroscopy method. From the determinations of this study, the C The widespread use of face masks reduces the spread of SARS-CoV-2 by minimizing the excretion of respiratory droplets from asymptomatic contaminated individuals or individuals who have not yet manifested symptoms ,2. This 2 concentrations ranging from 2150 to 2875 ppm and no differences were observed among the types of face masks tested. According to the literature, these concentrations have no toxicological effect [Guidance about mask wearing has varied from country to country, and some health bodies, including the World Health Organization, have modified their advice over time ,2,11. Sol effect ,11. Howe2, heart rate, respiration rate, and oxygen saturation on instructor pilots with relatively high CO2 concentrations  detected. Michelle S. M. Rhee et al., 2021 [2 increases with face masks but remains below short-term National Institute for Occupational Safety and Health limits.Another study    investig005  inveIn plants, ethylene is a major plant hormone mediating developmental processes and stress responses to stimuli, such as infection ,16. In hThe existing breath detection techniques designed to analyze air samples containing various volatile organic compounds include GC-MS/PTR-MS techniques, electronic nose (e-nose)  and infrCompared with non-spectral procedures, such as catalytic combustion, semiconductor gas sensing and electrochemical methods, there is no chemical reaction and no consumption of detection materials in the photoacoustic spectroscopy gas detection process, which makes the safety, stability, and life of a gas detection system highly improved . CompareThis study aimed to determine the concentration of ethylene and carbon dioxide from human respiration before and after one hour of wearing surgical face masks using photoacoustic spectroscopy gas detection technology.This research assesses the effect of wearing surgical face masks using laser photoacoustic spectroscopy gas detection in the evaluation of ethylene and carbon dioxide.Taking into account that the number of the absorbing molecules is proportional to the amplitude of the signal, ethylene and carbon dioxide emission was established in the case of 4 healthy volunteers aged between 31 and 41 years. −1. To improve the quantification of ethylene and carbon dioxide absorption coefficients, an adjusted process was adopted.A chemical compound that retains laser radiation is stimulated to a higher quantum condition causing a reduction in laser light intensity, which can be directly quantified via absorption spectroscopy ,31,32. T−3 cm−1atm−1 at 9P(18) laser transition.To determine gas absorptions, it is imperative to calibrate the resonant cell with a well-known gas compound and to establish the linearity of the detector with the concentration of the known gas over orders of magnitude. The linear responses of the cavity for the low detection limit of carbon dioxide and ethylene in literature is reported ,33. For −1atm−1 at the 10P(14) line of the laser.In the case of the ethylene absorption coefficients determination see a at the 2 laser was set at specific lines; first on the 9P (18) line: 9.53 μm where carbon dioxide showed a powerful assimilation, equivalent to an absorption coefficient of 3.01 × 10−3 cm−1atm−1 and then on the 10P (14) line 10.53 μm, where ethylene showed a powerful assimilation, equivalent to an absorption coefficient of 30.4 cm−1atm−1.All the measurements were made at the resonant cell pressure close to the atmospheric one and room temperature. So long as the absorption factors of carbon dioxide and ethylene at the separate laser wavelengths were specifically estimated ,33, the Below, in For the carbon dioxide and ethylene detection, we use software that records on different panels both the laser power, photoacoustic signal, and trace gases concentration.2. The participants were well-informed about the purpose and demands of the experimental study before giving their written consent to participate in this research. Information was asked regarding age, body weight and body height, time, and nature of the last meal and drink, recent exercise activity, medication, and smoking status.A total of 4 participants  were included in the study. The age ranged from 31 to 41 years old with 35 ± 7.6 yr, BMI 22.52 ± 4.2 kg/mThe breath was analyzed for a single subject with 4 samples per day for a period of 2 months. The participants were non-smokers, non-alcoholics, non-renal, non-diabetic, and without chronic mental or physical health problems, and without any recent antibiotic therapy. Prior to the analysis of the breath, the participants were asked to avoid the following for at least 6 h before, or at any time during, the breath sample collection: alcohol and coffee, food or beverages, and to refrain from exercise in the morning. On the day prior to the test, products such as onions, leeks, eggs, and garlic should be avoided.2 levels . The results for the carbon dioxide  on indiv2 increases with face masks when compared to controls. Otmar Geiss [2 concentration increase of 725 ppm, whereas Dattel et al. [2 concentration around 45,000 ppm. Similar to Otmar Geiss [2 increases with face masks but remains below short-term National Institute for Occupational Safety and Health limits.A number of papers ,13,14,34ar Geiss ,34 reporl et al. ,33, detear Geiss ,33 and Dar Geiss ,34, Mich2 at lower concentrations  has no toxicological effects while at higher concentrations  it causes the development of hypercapnia and respiratory acidosis  [2 concentrations between 2 and 3%  are known to produce sweating, headache and dyspnea  [2 concentrations are at 5% , mental depression may occur within several hours  [2 concentrations measured in this study are all far below these threshold values and range between 368 ppm  and 454 ppm .Inhaled COs, 2020) ,35. A coss 2020) ,36. Inhass 2020) ,36,37. I2 concentrations. As a vasodilator, the effect on people prone to headache has also been discussed. For example, Lim et al. 2006 [2 concentrations. Satish et al. 2012 [2 concentrations (approximately 2500 ppm) have the potential to affect decision-making.Carbon dioxide-related health-symptoms have been observed at concentrations above 1000 ppm and these include drowsiness and loss of attention  . A portial. 2006  administal. 2012  suggesteIn all these studies found in the literature ,38,39,40The ethylene concentrations measured in this study, similar to the carbon dioxide concentrations, are all far below the threshold values and range between 0.019 ppm  and 0.023 ppm , with a 0.004 ppm increase after one hour of wearing a surgical face mask.Face masks are essential components of personal protective equipment for health care workers in hospitals and public civilians alike. Notably, masks are helpful in preventing illness in healthy persons and preventing asymptomatic transmission, especially in a global pandemic. Although their use is now mandatory in many states due to COVID-19, they were generally used in hospitals and operating rooms even before the current pandemic. They do, however, increase both the carbon dioxide and ethylene concentrations behind them, as evidenced by this study. Although the results show that gases concentrations from the participant’s respiration increases after wearing a surgical mask for an hour, they remain below short-term National Institute for Occupational Safety and Health limits ,38,39,40The research was devoted to verifying the possibility that gases from respiration could build up in the mask over time, causing medical issues related to the respiratory system.2 laser photoacoustic spectroscopy method with respect to CO2 laser frequencies [So, the study evaluated the carbon dioxide and ethylene gases found in human respiration before and after wearing a surgical face mask using the COquencies ,41,42,43We used a specially designed sampling kit to collect a clean breath air sample with “alveolar” breath, which comes from the lungs, where gaseous exchange between the blood and breath air takes place.The exhaled air is a heterogeneous gas, and for a healthy individual, the first part of an exhaled breath, roughly 150 mL, consists of “dead-space” air from the upper airways (such as the mouth and trachea), where air does not come into contact with the alveoli of the lungs. The following part of a breath, about 350 mL, is “alveolar” breath, which comes from the lungs, where gaseous exchange between the blood and breath air takes place. For volatile organic compounds (VOCs) exchanged between the blood and alveolar air, the dead-space air is a “contaminant”, diluting the concentrations of VOCs when breath air is collected.For the identification of carbon dioxide and ethylene presence, all the participants’ respiration samples were collected in 0.75 L bags coated with aluminum and designed to accumulate and keep (for a maximum of 6 h) multiple respirations . The parThe photoacoustic spectroscopy gas detection used for the evaluation of the carbon dioxide and ethylene is summarized in 2 laser, a lens, a chopper, a photoacoustic resonant cell, a powermeter, a lock-in amplifier, an acquisition panel, and a data-processing computer. The experimental detection system also includes a gas handling system with an essential role in the control of the studied gases, but it can also perform other actions described in detail in other articles [The photoacoustic spectroscopy gas detection arrangement consists of a COarticles ,42,43,44articles ,44.2 laser photoacoustic spectroscopy gas detection system uses an adjustable laser and an H-type resonant cylindrical cell where the gas is detected. The laser beam enters the cylindrical cell after modulating and focusing. Similarly, to the photoacoustic cell, the microphone plays an important role in photoacoustic spectroscopy gas detection, and how to improve the sensitivity of the microphone has always been a hot topic.Basically, the CO2 laser radiation power is comparable to the molecular absorption coefficient of the analyzed gas sample at a CO2-used laser radiation wavelength. The microphones turn the acoustic signal into an electrical signal that is detected by a lock-in amplifier with a role in setting the chopper frequency and providing the photoacoustic signal. After passage through the stainless steel and Teflon cylindrical cell, the power of the laser beam is measured by a laser radiometer. Its digital output is introduced in the data acquisition interface module together with the output from the lock-in amplifier. The signals are gathered by an acquisition card and the data are recorded by a computer interface. The number of the absorbing molecules from the cylindrical cell is proportional with the amplitude of the photoacoustic signal. All experimental data are processed and stored by a computer. The frequency stabilized, line tunable CO2 laser emits in the 9.2–10.8 µm range, where a large number of gas molecules possess a high absorption coefficient. The light beam was modulated with a high quality, low vibration noise, a variable speed between 4 and 4000 Hz, and a mechanical chopper with a 30-slot aperture, operated at the appropriate resonant frequency of the H-type resonant cylindrical cell (564 Hz). The laser beam diameter is typically 5 mm at the point of insertion of the chopper blade and is nearly equal to the width of the chopper aperture. An approximately square waveform was produced with a modulation depth of 100% and a duty cycle of 50% so that the average power measured by the powermeter at the exit of the photoacoustic detector was half the cw value. The diverging infrared laser beam is converged by a ZnSe focusing lens (f = 400 mm). In this way, a slightly focused laser beam is passed through the resonant cylindrical cell without wall interactions. The useful data are automatically recorded, in real time, via a Test Point acquisition card under computer control.In the experiments ,42,43,44The dedicated program employed to acquire the data and calculate the value of the gas concentration was implemented using the Test Point platform. Further characteristics about the experimental set-up can be noted in the papers previously published ,42,43,442 laser photoacoustic spectroscopy gas detection method.Careful investigations of the carbon dioxide and ethylene concentrations of participants wearing surgical masks were performed using the COOur study demonstrates a non-significant increase in end tidal carbon dioxide and ethylene concentrations among participants while donning a surgical mask for one hour. According to the literature, these concentrations have no toxicological effect when inhaled. Therefore, there should not be concern for their regular day-to-day use. However, concentrations between 1000 ppm and 10,000 ppm can cause undesirable symptoms, such as fatigue, headache, and loss of concentration. This may be relevant for those segments of the population who are required to wear face masks over prolonged periods of time, such as students, bus drivers or cashiers, as well as persons with respiratory diseases. Although we recommend widespread mask usage, particularly during a respiratory disease pandemic, carbon dioxide and ethylene toxicity might complicate its use. Citizens should be provided regular opportunities to take breaks from the mask. A system to report symptoms related to mask use will be beneficial. A potential remedy is to change the surgical mask more often or designate an area to remove masks temporarily to mitigate any physiological effects associated with its use.In summary, the current work was carried out by implementing a methodology that assured better conditions to measure real concentrations of carbon dioxide and ethylene from human respiration. 2 laser photoacoustic spectroscopy gas detection method can be a new and easier method to assess molecules, which may contribute to a better understanding of the effect of wearing surgical face masks on gas detection from respiration. The implication of elevated carbon dioxide and ethylene levels with the long-term use of surgical masks needs further studies.The CO


Using the conventional rates  of urea as a surface application to produce 131.4, 257.2, and 212.1 Tg of wheat, maize, and rice resulted in 17–33 Tg, 7–10 Tg, and 6–87 Gg of AP, GWP, and AEP, respectively. For their balanced effect on reducing AP, GWP, and AEP while maximizing yields, inhibitors, and subsurface application could be set as the best mitigation strategies in wheat production. Inhibitors usage and biochar are strongly recommended strategies for sustainable production of maize. None of the investigated strategies had a balanced effect on rice yield and the environment, thus new mitigation technologies should be developed.Cropland reactive nitrogen losses (Nr) are of the greatest challenges facing sustainable agricultural intensification to meet the increases in food demand. The environmental impacts of Nr losses and their yield responses to the mitigation strategies were not completely evaluated. We assessed the environmental impacts of Nr losses in China and coupled the efficiency of mitigation actions with yield responses. Datasets about Nr losses in China were collected, converted into potentials of acidification (AP), global warming (GWP), and aquatic eutrophication (AEP), and analyzed by a meta-analysis program. Results showed that producing 1 Mg of rice grains had the highest AP (153 kg acid equiv.), while wheat had the highest GWP and AEP (74 kg CO China is the main contributor to these crops' production by 17.9, 22.9, and 28.1%, respectively  in the atmosphere, which is then rained out (acid rain)  and forms soluble (NH4)2 SO2 in atmospheric water vapor, which is the main source of N deposition  is a long-lived greenhouse gas and stratospheric ozone-depleting compound with an atmospheric lifetime of 116 ± 9 years  due to their broad coverage and capacity to carry out intricate search strings. The keywords  were used to screen databases. More than 600 papers were screened and 204 papers were selected (Data S1) with 1,893 datasets. These datasets were assessed for their relevance to the current study by the inclusion of the following criteria:Experimental details include study location, type, setup year, replicates number, crop type , climatic conditions (air temperature and precipitation), and soil parameters . Field and lysimeters studies only were included in the meta-analysis, while watershed-scale or modeling calculations and lab-based studies were excluded , rate and application depth, irrigation and amendments including biochar, inhibitors  thiophosphoric triamide dicyandiamide), and mulching.3, N2O, and −1, grain yield (Mg ha−1), standard deviation (STD), and standard error (SE).Accumulative seasonal Nr losses  via i which represents NH3, N2O, or −1). Then, Nr losses were converted into AP  12).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


In this brief report, we compare the effectiveness and safety of intermittent theta burst stimulation (iTBS) and conventional 10 Hz repetitive transcranial magnetic stimulation (rTMS) in patients with methamphetamine use disorder (MAUD). Our study suggests that iTBS would also reduce drug craving in patients with MAUD just as the 10 Hz; thus, there may be no difference in treatment effects between these two methods.n = 10) or 10 Hz (n = 10) groups for 12 treatments. Cue-evoked cravings, anxiety, depression, and withdrawal symptoms were measured at baseline before the first treatment, and post-tests after days 10, 15, and 20.In total twenty male methamphetamine (MA) addicts were randomly assigned to iTBS (The results showed that iTBS and 10 Hz treatment had similar effectiveness in reducing cue-induced craving in male addicts for MA. Both 10 Hz and iTBS improved withdrawal symptoms of patients with MAUD.Intermittent theta burst stimulation may be similar in effectiveness as 10 Hz in treating patients with MAUD. The clinical usefulness of rTMS could be improved substantially because of the increase in its capacity, cost, and accessibility. Importantly, the effectiveness of rTMS in the treatment of patients with MAUD is not yet proven, and should be tested in the large double-blind sham-controlled studies. Methamphetamine use disorder (MAUD) can cause serious social problems. It is well-accepted that patients with substance use disorder (SUD) experience high cravings and high-relapse rates. Currently, available treatments for MAUD mainly include an extension application of deep electrical stimulation therapy in the human brains based on animal optogenetic and electrical stimulation, which remain to be proven, because of the invasive nature and high price. In addition, target sites have not yet been identified. One of the most widespread addiction rehabilitation treatments in China is physical isolation, which combines physical rehabilitation and psychological counseling, but lacks targeted brain science techniques; implementation of psychological counseling requires a long period of time and extensive counselor experience. Its promotion is limited under existing conditions in China. Non-invasive brain stimulation techniques, such as repetitive transcranial magnetic stimulation (rTMS), may be a more scientifically sound option. rTMS induces sustained changes in the brain regions through high-intensity, focused pulsed magnetic fields , 2. AddiRepetitive transcranial magnetic stimulation is approved by the U.S. Food and Drug Administration (FDA) for the treatment of major depression , migrainThe effectiveness of iTBS has been confirmed in psychiatry and neurology –30. Meanhttp://www.chictr.org.cn; no. ChiCTR17013610). All the subjects signed informed consent forms before the experiment and participated voluntarily. The 20 MA participants were randomly assigned to either 10 Hz (n = 10) or iTBS (n = 10) in a 1:1 ratio using a simple randomization procedure.This study was a randomized, parallel-controlled case study. In total, twenty male MA addicts aged 24–53 years were recruited from the Gongchen Addiction Rehabilitation Center in Hangzhou, Zhejiang Province. Inclusion criteria included the use of MA . Exclusion criteria was other drug use, infectious disease, sleep deprivation, history of epilepsy or stroke, history of mental illness, metal implants in the brain, cochlear implants, increased intracranial pressure, traumatic brain injury, brain tumor, encephalitis, cerebrovascular disease, cerebral metabolic disease, pacemakers, history of heart disease, illiteracy, and previous rTMS treatment. Experimental procedures were approved by the Ethics Committee of Nanjing Normal University in accordance with the Declaration of Helsinki and the trial was registered in the Chinese Clinical Trial Registration Center , which quantitatively assesses craving in patients with MAUD; the mood scales for assessing subjects' withdrawal symptoms were Self-Rating Anxiety Scale (SAS) , Self-RaCraving score is an important factor in cue-induced addictive behavior and drug relapse. In our study, we asked drug users to watch a 5 min video of MA use in a relaxed state, and then, we assessed cue-induced craving scores using the VAS, with scores ranging from 0  to 100 (very much).In this study, the stimulation protocol was 10 Hz or iTBS, as described in previous studies , 33. Thet-test was used to compare the differences in demographic variables between the 10 Hz and iTBS groups. This study used a mixed experimental design of 2 (group: 10 Hz and iTBS) × 4 . We used two-way repeated measures ANOVA to compare the changes in craving, SAS, SDS, and MA withdrawal scores over time, between the two groups. The statistical significance threshold was set at p < 0.05.We analyzed the data from this study using IBM Statistical Product and Service Solutions (SPSS 19.0). An independent samples t-tests showed that there were no differences in demographic characteristics such as age , years of MA intake , maximum MA intake , and monthly MA intake  between the 10 Hz and iTBS groups.F = 46.944, p < 0.001, Post-hoc tests showed that 10 Hz reduced craving at day 10 , day 15 , and day 20  relative to baseline . Similarly, iTBS significantly reduced craving among MA addicts on day 10 , day 15 , and day 20  relative to baseline . The group main effect was not significant  .F = 12.26, p < 0.001, post-hoc tests, compared with the baseline , and the results showed that 10 Hz did not improve the anxiety of MA addicts on the 10th day , but improved the anxiety on the 15th day  and day 20  to some extent; however, relative to baseline , iTBS did not have this effect on day 10 , day 15 , and day 20 . The group main effect was not significant  .F = 4.20, p < 0.05, post-hoc tests, compared with the baseline , the results showed that depression in patients with MAUD was not improved by 10 Hz on the 10th day  or the 15th day , but improved to a certain extent on the 20th day . While relative to baseline , iTBS did not have this effect on day 10 , day 15  or day 20 . The group main effect was not significant  .F = 9.77, p < 0.001, Post-hoc tests were performed and compared with baseline ; the results showed that the withdrawal symptoms of MA addicts at 10 Hz did not improve on day 10  or day 15 , whereas at day 20 , there was a certain degree of improvement relative to baseline . iTBS showed no improvement on day 10  or day 15 , but there was some improvement on day 20 . The group main effect was not significant  .In total, three participants  reported mild dizziness or scalp pain after the first two sessions. The symptoms were relieved within 1.5 h. None of the subjects dropped out of the study due to adverse reactions. In general, both 10 Hz and iTBS reduced the cue-induced craving of male addicts for MA. In total 10 Hz or iTBS could improve withdrawal symptoms in patients with MAUD.This study suggests that iTBS is similar in effectiveness as 10 Hz in reducing cravings for MA addiction. Furthermore, there was no difference between the two stimulation forms for treating patients with MAUD. Both forms of rTMS (10 Hz and iTBS) can effectively reduce cue-induced cravings in patients with MAUD, which is consistent with the conclusions of previous studies –7, 24. TAlthough this study has certain advantages, it also has several limitations. First, the study did not design a sham group and could not properly eliminate time or placebo effects. Second, the treatment time for iTBS participants in each session was much shorter than that of the 10 Hz group, which may have led to a specific effect of time with iTBS. Third, we lacked MRI-guided neuronavigation in this study; although this method is not feasible or cost-effective for most studies conducted in addiction rehabilitation centers. As a reference, a previous study showed that in a similar experiment, BeamF3  could achieve the same stereotactic target as MRI . Fourth,In conclusion, we found that iTBS may have similar therapeutic effect compared with 10 Hz in patients with MAUD. Typical iTBS treatment  takes 5–10 min, while 10 Hz takes 15–20 min. Therefore, the number of patients with MAUD treated with each iTBS protocol per day can be increased by more than two-fold. In a broader sense, iTBS could have a more positive impact on the effects of enhancing treatment capacity, including improving treatment pathways, and reducing waiting times, thereby helping more patients with MAUD in addiction rehabilitation centers to benefit from the advantages of TMS and help physicians treat more patients.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.http://www.chictr.org.cn; no. ChiCTR17013610). The patients/participants provided their written informed consent to participate in this study.The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by Ethics Committee of Nanjing Normal University (2017-004) and was registered in the Chinese Clinical Trial Registration Center  and the National Science Foundation of China (Grant No. 81702230).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


To evaluate the efficacy and safety of oropharyngeal administration of colostrum (OAC) in preterm infants.We searched Embase, MEDLINE, Cochrane Central Register of Controlled Trials (CENTRAL), Cumulative Index to Nursing and Allied Health Literature (CINAHL), and the website of the clinical trials, search time was from the establishment of the databases or websites up to 1 February 2022. Preterm infants with gestational age (GA) ≤ 32 weeks or birth weight (BW) ≤ 1500 g were taken as the participants, collect randomized controlled trials (RCTs) of comparing OAC and placebo or no intervention in preterm infants. Two researchers independently screened the literature, extracted the data, and evaluated the quality of the literature, and we adopted Review Manager 5.3 software for meta-analysis.n = 1,173) were included in the review. A meta-analysis showed significant difference in the incidence of necrotizing enterocolitis , late-onset sepsis , ventilator-associated pneumonia , the time to reach full enteral feeds  = −3.40, 95% CI = −3.87 to −2.92), duration of hospital stay , and the rate of weight gain  between the colostrum group and control group. Meanwhile, researchers found no significant difference between the colostrum group and control group in the incidence of bronchopulmonary dysplasia , intraventricular hemorrhage , periventricular leukomalacia , retinopathy of prematurity , and patent ductus arteriosus .In total, 11 RCTs . Therefore, OAC can be used as part of routine care for preterm infants. It was found that a strategy of OAC in very preterm and extremely preterm infants did not decrease the composite primary outcome of death, LOS, or NEC , lactoferrin, bifidus factor, and lysozyme, which can regulate immunity, anti-infection, and sterilization . With th, or NEC . In view(1) Types of participants: Preterm infants with BW ≤1,500 g or gestational age ≤32 weeks were included in the review.(2) Types of studies: We included all published clinical randomized controlled trials (RCTs), and articles are written in English.(3) Types of intervention: The subjects in the colostrum group were given OAC, and the subjects in the control group were given placebo or routine care.(4) Types of outcome measures: Describe at least one primary or secondary outcome index set in this systematic analysis.(1) Non-human studies, reviews, commentaries, editorials, abstracts, letters, and so on.(2) Included subjects were literature on severe congenital malformations.(3) Republished literature.(4) Literature with unclear data description, wrong statistical methods, or data provided that cannot be analyzed by meta-analysis.(1) Incidence of NEC.(2) Incidence of LOS.(3) Death before discharge to home.(1) Time to reach full enteral feeds.(2) Duration of hospital stay.(3) Incidence of bronchopulmonary dysplasia (BPD).(4) Incidence of ventilator-associated pneumonia (VAP).(5) Incidence of IVH (grade ≥3).(6) Incidence of periventricular leukomalacia (PVL).(7) Incidence of retinopathy of prematurity (ROP).(8) Incidence of patent ductus arteriosus (PDA).(9) Rate of weight gain (kg.d).We conducted a comprehensive search that included the Embase, MEDLINE, Cochrane Central Register of Controlled Trials (CENTRAL), Cumulative Index to Nursing and Allied Health Literature (CINAHL), and the website of the clinical trials, search time from the establishment of the databases or websites was up to 1 February 2022. We searched using Medical Subject Headings (MeSH) terms: “preterm infant”[MeSH] OR “premature infant” [MeSH] OR “Infant, newborn” [MeSH] OR “Infant” [MeSH] OR “infant, low birthweight” [MeSH] OR “Infant, Very Low Birth Weight” [MeSH] OR “Infant, Extremely Low Birth Weight” [MeSH] OR “Infant, Extremely Premature” [MeSH] OR “Very premature infants” [MeSH] AND “Colostrum” [MeSH] OR “oropharyngeal administration.”We followed the standard processes recommended by the Cochrane Handbook for Systematic Reviews of Interventions. Two of the review investigators independently screened the titles and abstracts of all of the relevant studies identified by the search. After that, full-text articles were downloaded, read, and assessed using the confirmed criteria for exclusion or inclusion. Any disagreement between the researchers on the inclusion or exclusion of a paper was settled by consensus or by inviting a third reviewer.Two review authors independently extracted and compared data, in case of disagreement, the two reviewers settled it through consensus or by inviting a third reviewer. Last, we extracted the following data from each study.Literature basic characteristics .Participants .Intervention .Primary outcomes .Secondary outcomes , PVL, ROP, PDA, and rate of weight gain (kg.d).Two review authors according to the methods in the Cochrane Handbook for Systematic Reviews of Interventions independently assessed the risk of bias  of all included trials. Total of six domains were evaluated and categorized into low risk, high risk, and unclear risk. These six domains were random sequence generation, allocation concealment, blinding of participants and personnel, blinding of outcome data, incomplete outcome data, and selective reporting.I2 ≤ 50%, the fix-effect model was used for analysis, in which I2 < 25% means that there is no heterogeneity between the collected documents, and when 25% ≤ I2 ≤ 49%, it means that the heterogeneity between the collected documents is low. When I2 > 50%, it indicates that there is heterogeneity in the collected literature, of which 50% ≤ I2 ≤ 74%, it means that there is moderate heterogeneity between the collected documents, and when I2 > 75%, it means that the heterogeneity between the collected documents is high, the source of heterogeneity was further analyzed. After excluding the influence of obvious clinical heterogeneity, the random effects model was used for analysis. The relative ratio (RR) was used as the effect index for the counting data, and the standardized difference (SMD) was used as the effect index for the measurement data. For each effect quantity, given its point estimate and 95% confidence interval (CI). p < 0.05 was statistically significant.A meta-analysis was performed using the Review Manager (version 5.3). Firstly, the heterogeneity of the literature was tested. When Initially, 156 records were identified through database searching and other sources, 79 duplicate studies were excluded, and 49 were excluded after reading the title and abstract. Then, 17 were excluded after full-text reading, i.e., 7 non-RCT studies, 3 unable to obtain the full text, and 7 for other reasons. Finally, 11 RCTs met the inclusion criteria of this review. Among them, 581 in colostrum group and 592 in control group, a total of 1,173 preterm infants were included . The stuWe adopted the methods in the Cochrane Handbook for Systematic Reviews of Interventions to assess the risk of bias  of all included studies. The results of the risk of bias assessment are shown in I2 = 2%, p = 0.42) suggested that there was homogeneity among the studies, therefore, the fix-effect model was used for statistical analysis. A meta-analysis showed that there was significant difference in the incidence of NEC between the colostrum group and control group .Necrotizing enterocolitis is a common serious gastrointestinal disease in newborns. The main clinical manifestations are abdominal distension, vomiting, and bloody stool. All included studies –14 reporI2 = 28%, p = 0.18). A meta-analysis showed that there was significant difference in the incidence of LOS between the colostrum group and control group .Late-onset sepsis is a serious infectious disease in the neonatal period. The systemic inflammatory reaction is caused by pathogens that invade the blood of newborns and grow, reproduce, and produce toxins. In total, 11 studies –14, withI2 = 0%, p = 0.60) suggested that there was homogeneity among the studies. A meta-analysis showed there was no significant difference in the rate of death before discharge to home between the colostrum group and control group .In total, 8 studies –8, 12–14I2 = 92%, p < 0.00001). A meta-analysis showed there was significant difference in the time to reach full enteral feeds between the colostrum group and control group .In total, 7 studies , 11, 14 I2 = 94%, p < 0.00001). A meta-analysis showed that there was significant difference in duration of hospital stay between the colostrum group and control group .In total, five studies –6, 8, 14I2 = 0%, p = 0.55) suggested that there was homogeneity among the studies, therefore, the fix-effect model was used for statistical analysis. A meta-analysis showed that there was no significant difference in the incidence of BPD between the colostrum group and control group .Bronchopulmonary dysplasia is a common complication of premature infants. The current definition is that premature infants still need oxygen at 28 days after birth or 36 weeks after correcting gestational age. In total, 7 studies , 13 repoI2 = 0%, p = 0.41) suggesting that there was homogeneity among the studies. A meta-analysis showed that there was significant difference in the incidence of VAP between the colostrum group and control group .Ventilator-associated pneumonia is the main acquired infection in NICU, which is pulmonary inflammation 48 h after mechanical ventilation. In total, 5 studies , 13 repoI2 = 13%, p = 0.33) suggesting that there was homogeneity among the studies. A meta-analysis showed that there was no significant difference in the incidence of IVH (grade ≥3) between the colostrum group and control group .Intraventricular hemorrhage is a common type of intracranial hemorrhage in preterm infants, and it is also one of the main causes of death and disability in preterm infants. In total, 4 studies , 7, 13 rI2 = 0%, p = 0.56). A meta-analysis showed that there was no significant difference in the incidence of PVL between the colostrum group and control group .White matter injury is one of the unique forms of brain injury in premature infants. The most serious outcome is PVL, which causes neurological sequelae in children. In total, 2 studies , 8, withI2 = 0%, p = 0.88). A meta-analysis showed that there was no significant difference in the incidence of ROP between the colostrum group and the control group .Retinopathy of prematurity is an important disease that leads to blindness or severe visual impairment in preterm infants. In total, 5 studies , 8, 13, I2 = 0%, p = 0.47) suggested that there was homogeneity among the studies. A meta-analysis showed that there was no significant difference in the incidence of PDA between the colostrum group and the control group .Patent ductus arteriosus is one of the common congenital heart diseases and one of the main causes of heart failure in premature infants. In total, 2 studies , 5 reporI2 = 98%, p < 0.00001). A meta-analysis showed that there was significant difference in the rate of weight gain (kg.d) between the colostrum group and the control group .Totally, two studies , 6 reporvia the oropharyngeal route, beginning within 24 h after birth and continued until oral feeds were initiated, using a 1 ml syringe (without needle) with its tip placed between the cheek and the gum on either side  or sterile water (control) every 3 h from the day of life 2–7 . For then = 48), receiving 0.2 ml of oropharyngeal mother's milk every 4 h for the first 15 days of life, and a control group (n = 52), not receiving oropharyngeal mother's milk. At the end of the study, two newborns in each group were diagnosed with NEC (Bell stage ≥ 2). The results showed that there was no significant difference in the incidence of NEC (Bell stage ≥ 2) between the two groups  infants, the researcher compared oropharyngeal administration of mother's milk practice  with regular gavage feeding. It was found that oropharyngeal administration of mother's milk practice did not affect the incidence of neonatal mortality . VAP is The main conclusions of this meta-analysis are inconsistent with those of some previous meta-analyses and observational studies. The reasons may be as follows: (1) with the increasing research on OAC in recent years, medical staff is more proficient and standardized in the operation process of OAC; (2) most of the studies included in this meta-analysis are in recent 3 years, and the sample size is large, which has a great impact on the overall outcome; (3) intervention methods are slightly different: among the 11 included studies, 10 studies used oral drip to give breast milk, this method is simple to operate, but it may also damage the oral mucosa due to improper operation or reduce the ejection of colostrum due to the sucking action of preterm infants; 1 article used oral apply to give breast milk, this method is also very simple to operate, but the actual use amount cannot be estimated because the cotton swab will absorb most of the breast milk. (4) The control group was dripped or applied with different substances, such as normal saline, sterile water, nothing, and only routine care without any substances. Similarly, this study also has some limitations: (1) there are some differences in the start time and frequency of oral colostrum, and the amount of breast milk used for oral colostrum is also different, especially the duration of oral colostrum. Therefore, the intervention may not be entirely due to colostrum, which will affect the effect of colostrum oral application; (2) NEC and LOS are not unifactorial disorders and may be associated with a variety of factors, such as gestational age, BW, treatment received, and medical status of the preterm infant. Therefore, the effect of colostrum oral application on short-term and long-term outcomes in preterm infants still needs to be confirmed by long-term, large-sample, multicenter, and well-designed randomized controlled blinded studies.In conclusion, OAC can reduce the incidence of NEC, LOS, and VAP in preterm infants, shortening the time to reach full enteral feeds and duration of hospital stay, and increasing the rate of weight gain (kg.d). Although OAC is a simple and economical nursing operation, there are still many problems in the specific operation, such as the way of using OAC, the frequency, and the amount of colostrum used. Therefore, the efficacy and safety of OAC still need to be further confirmed by more studies.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.MH, ChunlL, HM, YuhZ, ChunzL, and DS contributed to the research, design, and analysis. MH, ChunlL, HM, YuhZ, ChunzL, DS, YayZ, YanZ, and CX contributed to the interpretation of the data and the drafting of the manuscript. All authors contributed to the article and approved the submitted version.This research was funded by the Natural Science Foundation of Inner Mongolia Autonomous Region (2020MS08034).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.